RU2592370C9 - Agent based on dehydroepiandrosterone, method for use thereof - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.

SUBSTANCE: invention concerns an agent on basis of dehydroepiandrosterone (DHEA) for normalising balance of estrogen and androgen in body, characterised by that it contains micronised powder of dehydroepiandrosterone (DHEA) with particle size from 1 to 5 mcm, or its microemulsion with particle size from 2 to 5 mcm and natural chitosan biocompatible nanostructured gel containing DHEA from 5 to 10 mg in 1 ml. Agent is used for menopausal women, as well as for percutaneous administration for elderly.

EFFECT: invention provides improved clinical values in patients of either sex.

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Description

The invention relates to substitution therapy and is aimed at creating biologically active, prophylactic and therapeutic agents in the field of endocrinology, andrology, gynecology, diabetology in the form of an external agent based on dehydroepiandrosterone (DHEA), which is an intermediate product of hormonal metabolism and helps to normalize the balance of estrogens and androgens in the body. In particular, the invention relates to compositions of compositions comprising chitosan in the form of a gel, and allows the administration of DHEA percutaneously or intravaginally effectively and conveniently compared to previously used ointments, plasters, gels.

Dehydroepiandrosterone (DHEA) is a phenanthrene-type steroid that is produced by the primates and the wild yam plant, whose structural formula is shown in FIG. 1. By primates, it is synthesized and secreted mainly by the adrenal cortex in sulfate form. Its daily production in humans reaches about 40 to 45 mg. Before puberty, it is synthesized in relatively small quantities, increasing with the onset of puberty and reaching a maximum level of 30 years. Then the level of its synthesis and entry into the blood begins to decline steadily at a rate of 60 ng / ml per year. At the age of 70 to 80 years, the concentration of DHEA in the blood decreases by 80%. The dynamics of age-related changes in DHEA in peripheral blood in men in the range of 22-112 years is shown in FIG. 2. DHEA circulates in blood vessels mainly in the form of DHEA sulfate (DHEA-C). The interconversion of DHEA / DHEA-C occurs at a high rate under the action of sulfatase and sulfotransferase. DHEA carries out its biological effect through multiple signaling pathways, including specific membrane receptors and, as a result of its intracellular transformation, into more biologically active derivatives of androgens and estrogens, in particular testosterone and estradiol (Dehydroepiandrosterone (DHEA) - a precursor steroid or an active hormone in human physiology - Traish AM, Kang HP, Saad F, Quay AT - J Sex Med 2011 8 (11) 2960-2982 guiz 2983 dol: 10.1111 / 5.1749-6109.2011.02.523.X). It is the change in the balance of androgenic and estrogenic activities that determines the beneficial effect of DHEA on obesity, cardiovascular disease, breast cancer, insulin sensitivity, cellular immunity, improving sexual function and physical strength, cognitive function, memory and Alzheimer's disease.

In experiments on old monkeys of M. Resus that reached the limit of biological age, studies with subcutaneous administration of DHEA at a physiological dose of 1 mg / kg body weight every 2 days for 1 month showed a sharp change in monkey behavior from passive to violent motor activity and increased nutritional motivation. According to the technique of motor-food conditioned reflexes, compensation of the level of DHEA in the body led to a significant increase in long-term and operative memory, which remains for 2 years after the termination of the introduction of DHEA. In monkeys, a coat that is lost with age is restored (Goncharov N.P., Katsiya G.V. et al. Effect of the neurosteroid dehydroepiandrosterone on the state of higher nervous activity of old monkeys. Human Physiology, 2014, Volume 40, No. 2, p. 1- 8).

Currently, a wide range of biological activity of DHEA has been proven. DHEA-based drugs are used to treat vaginal atrophy, hypogonadism, insufficient libido (US patent No. 5855548), obesity (US patent No. 5527788, 5795880, 5846962), osteoporosis (US patent No. 5776923, 5846960), uterine cancer, breast cancer, urine, colorectal cancer, diabetes (US Pat. No. 4,518,595), skin and autoimmune diseases, to enhance the immune response to vaccines (US Pat. No. 5,837,269), for general immunity enhancement (US Pat. No. 5,919,465), chronic fatigue syndrome, with reduced muscle mass (US patent No. 5756469), for diseases of the connective th tissue, with depression, for the treatment of bad habits (patent W09737663A1), as well as to increase life expectancy and improve the psychological state (US patent No. 5798347, No. 5807849). DHEA is widely used as a drug to increase its level in case of deficiency in the elderly. DHEA is synthesized in the brain de novo and is regarded as a neurosteroid for the prevention and treatment of Alzheimer's disease. DHEA increases the function of the central nervous system (Goncharov NP, Katsiya GV Neurosteroid DHEA and brain function. Journal of Human Physiology, 2013, Volume 39, No. 6, pp. 120-128).

Currently, various methods for the delivery of DHEA to the body are known: the oral method in the form of tablets, including sublingual, capsules, including gelatine, sublingual solutions, chewing gums, a method of application in the form of a cream or gel containing numerous chemical components, as well as a transdermal method in the form of a patch [US patent No. 4978532, No. 56869090].

However, in all of the listed forms of using DHEA, one of the main and significant problems is that to achieve the necessary biological and therapeutic effect, it is necessary to use sufficiently high doses (from 40 to 100 mg / day) due to the rapid metabolism in the liver of free DHEA to DHEA sulfate. before it enters the bloodstream, which is a consequence of low therapeutic bioaccumulation and a decrease in the effectiveness of currently produced and used drugs.

The use of a high initial dose is due to the very low solubility of DHEA in both water and oil. In a hydrophilic environment, crystalline DHEA has a tendency to sedimentation, coalescence of particles and aggregation, which is a certain obstacle to their effective resorption in the oral cavity or transfer to the body in case of percutaneous application. The use of oil or fat forms of various creams or artificial gels having a multicomponent chemical composition avoids the initial ingestion of DHEA into the liver with subsequent metabolism, but it does not reduce the high doses of DHEA applied during applications to achieve the required concentration of it in the blood also due to the imperfect composition of the products based on DHEA and its low degree of absorption (about 10% of the applied dose).

A similar and physiological effect, the developed and manufactured Androgel product (manufactured by Laboratoires BESINS INTERNATIONAL, France) contains, in addition to the testosterone steroid, a gel based on chemically synthesized components and up to 70% ethyl alcohol. A sufficiently large volume of application (5 ml of gel and 25 mg of testosterone) in this case, in addition to side effects with prolonged use, can have a damaging effect on liver tissue, kidneys and other organs and tissues.

The same disadvantages apply to the preparation for transdermal administration of DHEA - a cream with DHEA (manufactured by NOW Foods, Ltd., USA), and chosen by the authors as an analogue in which this multicomponent product along with DHEA contains 18 chemical and herbal ingredients. To achieve a sufficient therapeutic effect when using this drug, it is necessary to apply an application in an amount of at least 15 mg of DHEA.

Thus, the presented and other products based on steroids do not solve the problem of its low absorption and all the associated side undesirable effects due to the need to use a high initial dose and the presence of numerous chemical components in ointments and gels.

The claimed invention eliminates all of the above problems by creating an original composition (composition) of a transdermal product containing either micronized DHEA powder with a particle size of 1 to 5 μm or a DHEA microemulsion with a particle size of 2 to 5 μm with a nanostructured chitosan gel.

The composition developed by the authors has a number of significant advantages compared to existing means in the form of ointments or gels:

1. The composition of the developed composition does not include any additional chemical components, except for DHEA and biologically similar nanostructured chitosan with a moisture content of 95 to 99.5%, which ensures the absence of undesirable side effects (toxicity, allergies, irritation, etc.).

2. The composition of the developed composition and the method of its use make it possible to exclude the possibility of excessive formation of nonphysiologically overestimated estradiol and testosterone from DHEA, which may have undesirable side effects.

3. The composition of the developed composition and the method of its application allows us to solve the problem of bioaccumulation of DHEA, its bioavailability and significantly reduce the initial dose compared to products on the market (ointments, gels and capsules, tablets) containing DHEA in the range from 2 to 10 times more than in the composition developed by the authors.

4. The composition of the developed composition and the method of its application by percutaneous application can reduce the amount of DHEA used for application to the minimum physiological level (up to 5-10 mg per day) to compensate for its deficiency during long courses of administration, thereby eliminating its metabolic load on the liver.

Chitosan used in the composition of the composition is well known as a non-toxic biocompatible polymer, a derivative of chitin and is a widely used and commercially produced product by various manufacturers both in Russia and abroad. It is a linear polysaccharide consisting of randomly linked D-glucosamine and N-acetyl-D-glucosamine units. This is a glucose polymer in which the C-2 hydroxyl group is replaced by an N-acetyl amino group (NHCOCH ₃ ). In chitosan, an acetyl group is absent. Thus, chitosan is a deacetylated chitin, contains about 7% nitrogen and is structurally similar to cellulose. Chitin can be obtained from the shells of crab and other crustaceans in the form of an amorphous powder after processing the calcium carbonate shell and removing proteins. The most promising form of chitosan in the form of a nanostructured hydrophilic gel is obtained by further processing (Ohya et al., 1994; Yokohama et al., 1998; Kataoka et al., 2000; Preparation and application of chitosan nanoparticles and nanofibers. Li-Ming Zhao et al. Braz. J. Chem. Eng. vol. 28 no. 3 Sao Paulo July / Sept. 2011; RF patent No. 2428432).

A review by T. Klean and M. Thanou, “Biodegradation, Biodistribution, and Chitosan Toxicity,” published in the journal (Advanced Drug Delivery Reviews, 2010, v. 62, p. 3-11), shows the absence of any toxic effect of chitosan. Due to the presence of NH ₂ groups in its structure and good biocompatibility, chitosan is considered as a promising transport form for the delivery of drugs to the body. Due to the positive charge, chitosan has the most interesting property for using it as a penetration agent through the skin or mucous membrane.

Chitosan is approved and widely used in pharmaceutical and food production in Japan, Italy, Finland, the USA and Germany. Due to the positive charge of the molecule, he found application in the drug for stopping profuse bleeding due to the mechanism of electrostatic action on negatively charged red blood cell membranes (B.C. Schmid, G.A. Rezniczek et al. Amer J Obstetrics and Gynecology, 2013, (3) 225.el).

A comprehensively investigated claimed composition of DHEA with a chitosan nanostructured gel, despite its hydrophilic nature, showed remarkable delivery properties of the hydrophobic substance DHEA through the skin and mucous membranes and thereby dramatically reduced its therapeutic dose, increased bioavailability, absorption and provided the necessary concentration blood. The claimed composition for percutaneous application with low physiological doses (from 5 to 10 mg / day), in contrast to other high doses of DHEA (15 to 50 mg / day or more) used in other drugs, eliminates the metabolic load on the liver and, which is especially important , to exclude the possibility of the formation of non-physiologically overestimated estradiol and testosterone from DHEA, which can have undesirable side effects, thereby ensuring the most optimal compensation for menopause by both estrogens and androgens that are formed target tissues from the primary source of DHEA. Fundamental studies of Professor F. Labry on the process of DHEA metabolism and its intracellular transformation in target tissues into more biologically active estrogens and testosterone have shown the particular importance of the presence and control of appropriate enzyme systems that have a locally biological effect due to undesirable metabolism (F. Labrie, A. Belanger, Van Luu-the, et al. DHEA, The precursor of Androgens and Estrogens in Peripheral Tissues in the Human: Intracrinology. Dehydroepiandrosterone (DHEA) edited by M. Kalimi and W. Regelson 2000, p 299-342 ; F. Labrie et al. Serum steroid levels during 12 -week intravaginal dehydroepiandrosterone administration. The journal of The North American Menopause Society. 2009, 5, 897-906; Trivedi DP, Khaw KT Dehydroepiandrosterone sulfate and mortality in elderly men and women. J Clin Endocrin Metab, 2001, 80, 9: 4171 -4177). This is especially important for women during menopause and postmenopause, when the ovaries lose hormonal function and the main source of estradiol and testosterone is DHEA, which is produced by the adrenal glands. Its concentration in the peripheral blood of menopausal women is reduced, but the level of its secretion is sufficient for the intracellular formation of estradiol and testosterone in the peripheral tissues, especially in adipose tissue, and from there they enter the systemic circulation.

The use of the claimed composition DHEA / chitosan nano-gel for percutaneous application allows you to adjust and establish the necessary therapeutic level of DHEA in the blood, which, as shown previously (Trivedi DP, Khaw KT - Dehydroepiandrosterone sulfate and mortality in elderly men and women. - J Clin Endocrinol Metab, 2001 80, 9: 4171-4177; Morales AJ, Nolan JJ et al. Effect of replacement dose of dehydroepiandrosterone in men and women of advancing age. J Clin Endocrinol Metab, 1999, 78 (6), 1360-1367; Goncharov N. P., Katsiya GV The hormone of health and longevity, pp. 134-141. Adamant Publishing House, Moscow, 2012) is able to provide correction of the entire cluster of symptoms, characteristic x for menopause, including osteopenia and osteoporosis, loss of muscle mass, reduction and elimination of hot flashes, changes in the volume of adipose tissue, age-related changes in the skin, deterioration in cognitive function and memory, correction of the severity of diabetes 2 in its presence and Alzheimer's disease.

The above examples confirm the originality and novelty of the claimed composition DHEA in chitosan nanogel and the method of its application, but do not limit the scope of its application.

Percutaneous application of DHEA in chitosan gel

To evaluate the effectiveness of the use of the DHEA / chitosan nanogel composition compared with the oral method, percutaneous application was used as a replacement therapy. The initial therapeutic dose of DHEA in the composition developed by the authors was 5 times less than when administered orally. The pharmacokinetics of free DHEA was evaluated in both men and women aged 68 to 78 years. The determination of free DHEA and testosterone was carried out by its content in saliva, since this is a more convenient, faster and non-invasive method for determining steroid hormones compared to the methods of such analyzes by taking blood due to the fact that the concentration of free DHEA in saliva is highly correlated with the concentration of total DHEA in blood. In FIG. Figure 3 shows the dynamics of age-related changes in DHEA in peripheral blood in men in the range of 22-112 years of Hbc (Science for Health. Immuno Biological Laboratories IBL. Saliva Diagnostics www.salivadiagnostics.de).

Method for percutaneous application of DHEA in a chitosan gel: DHEA in a dose of 5 to 10 mg per 1 ml of gel is applied to the inner surface of the forearm or thigh with an area of 10 cm ² and rubbed into the skin with light rubbing for about 5 minutes. This ensures rapid transdermal entry of DHEA into the capillary and general bloodstream with a maximum concentration in 30 to 60 minutes.

Example 1

The man is 78 years old. Initial state: symptoms of age-related deficiency, including general and muscle weakness, increased fatigue by the end of the day, drowsiness, depression.

A composition of DHEA / chitosan nanogel with a particle size of chitosan of not more than 50 nm was used. As a result of daily application of the DHEA / chitosan gel composition at a dose of 10 mg in 1 ml to the skin surface, the symptoms of age-related deficiency significantly improved after 14 days, and after 30 days they returned to normal. The level of circulating DHEA has increased to the level characteristic of men aged 30 to 40 years. In parallel with the increase in DHEA, there was a slight increase in the concentration of free testosterone in saliva.

A comparative assessment of the pharmacokinetics of free DHEA after a single transdermal application of a 10 mg DHEA / chitosan gel composition and after a single oral administration of a DHEA capsule at a dose of 25 mg and against a background of chitosan gel without DHEA application showed that a transdermal application of 10 mg of DHA in 1 mg more than 1 DHA in 1 a high (from 75 to 228 %%) increase in the concentration of free form of DHEA in comparison with oral administration of 25 mg of DHEA. Dynamics of free DHEA in saliva after a single application of DHEA in chitosan gel (10 mg of DHEA in ml of gel); after a single oral administration of a DHEA capsule (25 mg) and the dynamics of free DHEA after application of a chitosan gel without DHEA is shown in FIG. four.

Example 2

The patient is 68 years old.

Initial state: pronounced deficiency of DHEA and testosterone, complaints of increased fatigue, depression and memory impairment. After 2 weeks of DHEA administration, fatigue and depressive symptoms decreased, and after two months there was an improvement in all indicators of age-related symptoms, including cognitive function.

The pharmacokinetics of DHEA and the testosterone formed from it was studied in the case of application of the DHEA / chitosan gel composition developed by the authors.

As a result of the application of a single application of the DHEA / chitosan gel composition, after 1 hour, the content of free DHEA increases by 15 times compared to the initial one. At the same time, a pronounced rise in free testosterone is detected in saliva. But its maximum values are recorded with a delay of about 2 hours compared with DHEA. After 24 hours, the level of testosterone in saliva exceeded the baseline by 95%, i.e. there is a compensation for its deficiency, which is characteristic of women during menopause. The dynamics of free DHEA in saliva depending on the dose of DHEA for percutaneous application of DHEA in women in the postmenopausal period is shown in FIG. 5. The dynamics of an increase in the level of free testosterone in saliva against the background of percutaneous application of DHEA in a postmenopausal woman with androgen deficiency is shown in FIG. 6.

The obtained pharmacokinetic data suggest that the claimed composition, consisting of DHEA and biologically similar chitosan in the form of a gel, in comparison with the known formulations and forms of tablets, capsules, creams and gels of a different composition, solves the problem of the initial high therapeutic dose, bioaccumulation and bioavailability using a lower initial dose from 2 to 10 times and can be the basis for the creation of prophylactic agents, dietary supplements and drugs instead th therapy.

Intravaginal administration of DHEA in chitosan gel

In cases of vaginal atrophy in women, the effectiveness of the effect of the DHEA / chitosan gel composition was studied with a decrease in the initial therapeutic dose of DHEA by 2-2.3 times compared with the dose if used. The average age of women ranged from 51 to 56.6 years, with surgical menopause lasting from 1 to 14 years, which is the most severe in its clinical manifestations. Surgical menopause develops rapidly with surgical bilateral removal of the ovaries, which is performed according to strict medical conditions.

Example 3

Diagnosis: Surgical menopause (1 year).

Menopausal syndrome. Atrophic vaginitis.

Prior to the use of therapy, vegetative-emotional tides were observed more than 10 times a day, high astheno-depressive manifestations (Green scale 30 points), low vaginal health index (VHI) ranged from 1 to 2 points, and the pH of the medium was more than 6.1. There is no elasticity, severe dryness, bleeding of the mucous membrane upon examination. Unfavorable level of microflora of the separated vagina: Lactobacillus - not detected, Enteroccocus faecalis 1 · 10 ⁷ , Escherichia coli - copious growth.

A 5 mg daily intravaginal administration of the DHEA composition was prescribed in 1 ml of chitosan gel for three months. As a result of the application, vegetative-emotional manifestations significantly improved. The Green scale decreased from 30 to 22 points, the vaginal health index increased from 1-2 to 4 points; bacteriological analysis showed a significant improvement in the microflora of the separated vagina: Lactobacillus sp.- 1 · 10 ⁵ , Enteroccocus faecalis 1 · 10 ⁵ , Escherichia coli 1 · 10 ⁴ . An additional positive effect on the normalization of urination was revealed.

Example 4

Diagnosis: Surgical menopause (14 years). Menopause syndrome, emotional-vegetative form. Atrophic vaginitis. Metabolic syndrome.

Initially, the woman was dominated by the emotional-vegetative form of menopause (hot flashes more than 10 per day), astheno-depressive symptoms. On the Green scale, the high severity of the manifestations of menopausal syndrome corresponded to 36 points. Urogenital manifestations (urinary incontinence, frequent urination), IVS corresponded to values from 2 to 3 points, pH was 5.8. On examination, weak elasticity, severe dryness, an inflamed surface, and bleeding were noted. Bacteriological analysis of the vaginal discharge revealed an unfavorable level of microflora: Lactobacillus - not detected, Enteroccocus faecalis 1 · 10 ⁷ , Staphyloccocus spp - 1 · 10 ⁴ , Escherichia coli - 1 · 10 ⁵ .

A daily intravaginal administration of the DHEA composition at a dose of 5 mg in 1 ml of chitosan gel was prescribed for three months, which allowed to obtain a significant improvement in health status. The number of hot flashes is reduced by 2 times (from 5 to 6 per day), normalization of urination is registered. The Green scale decreased 1.8 times to 20 points, and the IVS rose to 4 points, the pH was 4.8. Significant improvement in elasticity, lack of bleeding and normalization of moisture in the vaginal mucosa were noted. Bacteriological analysis showed the normalization of microflora: Lactobacillus - 1 · 10 ⁴ , Enteroccocus faecalis 1 · 10 ³ , Staphyloccocus spp 1 · 10 ⁴ .

Example 5

Diagnosis: Surgical menopause (3 years).

Menopause syndrome, emotional-vegetative form. Atrophic vaginitis. The Green scale - 28 points (astheno-depressive symptoms and urogenital manifestations dominate), the IVS indicator ranged from 2 to 3 points: pH is 5.6, elasticity is weak, severe dryness, the surface is not inflamed. According to the bacteriological analysis of the vaginal discharge: Lactobacillus - 1 · 10 ² , Enteroccocus faecalis - 1 · 10 ⁴ , Staphyloccocus spp - 1 · 10 ³ .

A daily intravaginal administration of the DHEA composition at a dose of 5 mg in 1 ml of chitosan gel was prescribed for three months, which allowed to obtain a significant improvement. By the second month, a decrease in the Green score from 28 to 11 points was noted, the IVS indicator increased to 4 points with a normalization of the indicator (pH 4.9), elasticity and humidity. Normalization of the vaginal mucosa was noted according to microbiological indications (Lactobacillus sp. - 1 · 10 ⁵ , Staphyloccocus spp - 1 · 10 ³ ).

Example 6

Diagnosis: Surgical menopause (3 years).

Menopause syndrome, emotional-vegetative form. Atrophic vaginitis. The initial indicator on the Green scale was 27 points (vegetative-emotional manifestations dominate). The IVS indicator was from 2 to 3 points: pH is 5.2, elasticity is weak, severe dryness, the surface is not inflamed. According to the bacteriological analysis of the vaginal discharge: Lactobacillus - 1 · 10 ² , Enteroccocus faecalis - 1 · 10 ⁶ , Corynebacterium amycolatum - 1 · 10 ³ .

A daily intravaginal administration of the DHEA composition at a dose of 5 mg in 1 ml of chitosan gel was prescribed for 3 months, which allowed to obtain a significant improvement. By the 2nd month, a significant decrease in points on the Green scale to from 27 to 14 points was noted. The IVS indicator increased to 4 points. At the same time, the pH was set at 4.4, normalized elasticity and humidity. Bacteriological analysis of the vaginal discharge showed normalization of the microflora (Lactobacillus sp. - 1 · 10 ⁵ , Enteroccocus faecalis - 1 · 10 ² ).

The presented results on the use of the claimed means based on DHEA / chitosan gel to eliminate the symptoms of vulvovaginal atrophy in menopausal women confirm a significant improvement in all clinical indicators. A decrease in the Green score and an improvement in the vaginal health index were established, which allows to obtain a significant positive result: the vaginal structure is restored, the condition of the vaginal microflora is improved and all subjective symptoms associated with menopause are improved.

Claims (3)

1. A tool based on dehydroepiandrosterone for normalizing the balance of estrogens and androgens in the body, characterized in that the product contains micronized dehydroepiandrosterone powder with particle sizes from 1 to 5 microns or its microemulsion with a particle size of 2 to 5 microns and a chitosan natural biocompatible nanostructured gel with DHEA from 5 to 10 mg per 1 ml. 2. The tool according to claim 1 is not more than 1 ml with a DHEA content of not more than 5-10 mg for daily vaginal administration to menopausal women. 3. The tool according to claim 1 is not more than 1 ml with a DHEA content of not more than 5-10 mg for daily percutaneous administration to the elderly.

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