KR20240022819A - Use of 4-hexylresorcinol for treating male sexual dysfunction or infertility caused by diabetes - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating diabetes-related male sexual dysfunction or male infertility, comprising 4-hexylresorcinol or a pharmaceutically acceptable salt thereof as an active ingredient. According to the present invention, 4- Since treatment with hexylresorcinol can suppress testicular atrophy and increase sperm production ability, 4-hexylresorcinol can be useful in preventing, alleviating or treating male sexual dysfunction or male infertility caused by diabetes. there is.

Description

{Use of 4-hexylresorcinol for treating male sexual dysfunction or infertility caused by diabetes}

The present invention relates to the use of 4-hexylresorcinol in the treatment of male sexual dysfunction or male infertility.

Diabetes is a chronic metabolic disorder characterized by hyperglycemia, and has recently shown rapid prevalence in modern society, threatening individual health. Diabetes' threat to health can also have a profound impact on the male reproductive system. Diabetes can also lead to decreased testosterone levels and seminiferous tubule diameter, decreased virility and spermatogenesis, abnormal sperm production, testicular atrophy, premature ejaculation, frigidity, and increased infertility. It causes complications related to male reproductive function, such as:

In general, male sexual dysfunction such as erectile dysfunction, seborrhea, and premature ejaculation are treated by injecting male hormones, but in the case of male sexual dysfunction due to diabetes, male hormone levels are sometimes measured in the normal range, so treatment is performed with male hormone drugs. may not be possible. In addition, long-term use of male hormones in excessive amounts may cause testicular atrophy, enlargement of the prostate, and excessive stimulation of hematopoietic cells in the bone marrow, causing an increase in red blood cells, leading to the possibility of cardiovascular disease due to cytocytosis. There may be side effects such as depression, sudden weight gain due to increased body moisture, worsening of cardiovascular diseases such as high blood pressure, and reduction of high-density cholesterol, which is good cholesterol. Additionally, if you continuously supplement with male hormones, the body's natural production ability may be suppressed, resulting in a decrease in sperm count and semen volume. Therefore, in the treatment of male sexual dysfunction caused by diabetes, there is a need for other treatments to replace male hormone drugs.

Another disease that can be caused by diabetes is male infertility. Male infertility is caused by a variety of factors, including hormonal problems including male hormones, testicular problems due to exposure to toxic substances, infection or trauma, sperm problems, and abnormal sperm discharge paths. Recently, it has been reported that male infertility shows a higher-than-average rate of increase in relation to modern lifestyles such as obesity, diabetes, smoking, and stress. In particular, in the case of diabetes, oxidative stress induced by high blood sugar increases the function of testicular steroid production and spermatogenesis. It can cause disability and accelerate testicular damage. This damage to testicular tissue is closely related to sperm motility, sperm concentration, and number, and leads to the deterioration of sperm quality. As with male sexual dysfunction, hormones are commonly used to prevent or treat infertility, but because the side effects caused by them are problematic, other treatments are required to replace them.

The purpose of the present invention is to provide a new use of 4-hexylresorcinol for preventing, improving or treating male sexual dysfunction or male infertility due to diabetes.

The present inventors confirmed that when 4-hexylresorcinol was treated in male rats with induced diabetes, testicular atrophy was suppressed and the expression of LGR-4 and testosterone increased in the testes, thereby improving male sexual function induced by diabetes. The present invention was completed after confirming that 4-hexylresorcinol can be effectively used in the treatment of disorders or male infertility.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating diabetes-related male sexual dysfunction or male infertility, comprising 4-hexylresorcinol or a pharmaceutically acceptable salt thereof as an active ingredient.

According to the present invention, treatment with 4-hexylresorcinol can suppress testicular atrophy and increase sperm production ability, so 4-hexylresorcinol can be used to prevent and alleviate male sexual dysfunction or male infertility caused by diabetes. Or, it can be useful for treatment.

Figure 1 confirms that testicular atrophy is suppressed when 4-hexylresorcinol is treated in male rats with induced diabetes.
Figure 2 shows that when 4-hexylresorcinol was treated in male rats with induced diabetes, the expression of LGR-4 was increased in the testes.
Figure 3 shows that when 4-hexylresorcinol was treated in male rats with induced diabetes, it was confirmed that atrophy of the testis itself was prevented, atrophy of sperm in the testis was prevented, and expression of LGR-4 and the male hormone testosterone in testicular tissue was increased. .

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating diabetes-related male sexual dysfunction or male infertility, comprising 4-hexylresorcinol or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, “4-hexylresorcinol” is a compound with the IUPAC name of 4-hexylbenzene-1,3-diol, having the following formula (1) It is a compound represented by:

[Formula 1]

In the present invention, the 4-hexylresorcinol can be used interchangeably with “4-hexylbenzene-1,3-diol” or “4-HR”.

In the present invention, the pharmaceutically acceptable salt of 4-hexylresorcinol is a concentration that has an effective effect that is relatively non-toxic and harmless to patients, and the side effects caused by this salt reduce the beneficial efficacy of 4-hexylresorcinol. It refers to an organic or inorganic addition salt that does not separate. For example, pharmaceutically acceptable salts may be acid addition salts formed using organic acids or inorganic acids, wherein the organic acids include, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, and malic acid. , maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid, aminooxy acetic acid, benzene. Sulfonic acid, p-toluenesulfonic acid, and methanesulfonic acid-based salts are included, and inorganic acids include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, and boric acid-based salts. Preferably, the pharmaceutically acceptable salt may be in the form of citrate or hydrochloride. Additionally, the pharmaceutically acceptable salt may be an alkali metal salt (sodium salt, potassium salt, etc.) and an alkaline earth metal salt (calcium salt, magnesium salt, etc.).

In the present invention, “male sexual dysfunction” may refer to all disorders related to sexual life, including decreased libido or penile curvature. In one embodiment of the present invention, the male sexual dysfunction may be erectile dysfunction, premature ejaculation, seborrhea, or retrograde ejaculation, but is not limited thereto.

In the present invention, “infertility” refers to male infertility, unless otherwise specified, and “male infertility” refers to infertility that occurs due to abnormal sperm formation. In one embodiment of the present invention, the male infertility may be azoospermia, oligospermia, astheno spermia, or tetrazoospermia, but is not limited thereto.

In the present invention, the testes are sperm-producing organs among the male reproductive organs, and perform an exocrine function of producing and secreting mature sperm and an endocrine function of synthesizing and secreting the male hormone testosterone. Therefore, if the testicles become inflamed or atrophied, abnormalities in spermatogenesis may occur, for example, the number and motility of sperm may decrease and the number of malformed sperm may increase.

According to the following examples, 4-hexylresorcinol can suppress testicular atrophy and therefore can be usefully used for the treatment of male sexual dysfunction or male infertility.

In the present invention, "LGR-4" refers to G protein-coupled receptor 4 (Leucine-rich repeat), a G protein-coupled receptor (GPCR) that plays an important role in various physiological functions. -means containing G-protein coupled receptor 4). Unlike common G-protein coupled receptors, LGR4 does not activate heterotrimeric G-proteins to transmit signals, but rather Wnt signaling, which is required for the development of various organs, including the liver, kidneys, intestines, bones, reproductive tract, and eyes. It functions to activate the path. LGR4 may also serve as a receptor for norrin (NDP), which activates the Wnt signaling pathway in peritubular myoid cells during spermatogenesis. The expression of LGR4 can be reduced in heavy metal pollution such as cadmium or metabolic diseases such as diabetes, resulting in bone loss and decreased skin regeneration (Wang M, Liu J, Zhu G, Chen X. Low levels of cadmium exposure affect bone by inhibiting Lgr4 expression in osteoblasts and osteoclasts. J Trace Elem Med Biol. 2022:127025.: Wang M, Yao S, He D, Qahar M, He J, Yin M, Wu J, Yang G. Type 2 Diabetic Mellitus Inhibits Skin Renewal through Inhibiting WNT-Dependent Lgr5+ Hair Follicle Stem Cell Activation in C57BL/6 Mice. J Diabetes Res. 2022; 2022: 8938276.). Since deletion of the LGR4 gene causes reproductive abnormalities in rats, LGR4 is thought to play a very important role in normal testicular development (Hsu PJ, Wu FJ, Kudo M, Hsiao CL, Hsueh AJ, Luo CW. A naturally occurring Lgr4 splice variant encodes a soluble antagonist useful for demonstrating the gonadal roles of Lgr4 in mammals. PLoS One. 2014 Sep 4;9(9):e106804.). When sleep disorders are induced in laboratory rats, sperm function is reduced, and the expression of LGR4 is reduced in these rats (Chen W, Guo X, Jin Z, Li R, Shen L, Li W, Cai W, Zhang G. Transcriptional alterations of genes related to fertility decline in male rats induced by chronic sleep restriction. Syst Biol Reprod Med. 2020;66(2):99-111.). Therefore, if the expression of LGR4 is reduced, abnormalities in sperm function or sperm formation may occur.

According to the following examples, 4-hexylresorcinol can increase the expression of LGR-4 in the testes, and therefore can be used to treat diseases associated with a decrease in LGR-4 and abnormal production of sperm, that is, male sexual dysfunction or male infertility. It can be useful for

In the present invention, "testosterone" is a steroid hormone produced in Leydig cells of male testes. In men, testosterone maintains sexual function, promotes sperm formation, and develops and maintains muscles. . When testosterone decreases in adults, infertility, decreased sexual function, erectile dysfunction, testicular atrophy, and decreased sperm motility occur.

According to the following examples, 4-hexylresorcinol can increase the expression of testosterone in the testes of laboratory rats with diabetes, and therefore, diseases caused by decreased expression of testosterone with diabetes, such as male sexual dysfunction or male infertility. It can be useful for the treatment of.

In the present invention, 4-hexylresorcinol (4-HR) may be included at a concentration of 0.1 μM to 1 mM. For example, 4-hexylresorcinol may be included in a concentration of 0.5 μM to 500 μM or 1 μM to 100 μM. In the above concentration range of 4-hexylresorcinol, the effects of suppressing testicular atrophy and increasing the expression of LGR4 or testosterone can be maximized.

The route of administration of the pharmaceutical composition containing 4-hexylresorcinol as an active ingredient may be administered through any general route as long as the drug can reach the target tissue, and may be administered to the subject orally or parenterally. You can. For example, intraperitoneal administration, intravenous administration, intraarterial administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration (including dermal application, instillation, and inhalation), transdermal administration, intrathecal, and epidural. Administration, intranasal administration, intraocular administration, intrapulmonary administration, intrarectal administration, intravaginal administration, etc. may be performed, but are not limited thereto. In addition, the pharmaceutical composition containing 4-hexylresorcinol as an active ingredient may be in any convenient pharmaceutical product form, such as tablets, powders, granules, capsules, oral solutions, solutions, dispersions, suspensions, syrups, It can be administered as a spray, suppository, gel, emulsion, patch, etc.

The pharmaceutical composition of the present invention may further include pharmaceutically or physiologically acceptable carriers, excipients, and diluents. In the present invention, the term “pharmaceutically acceptable carrier, excipient, and diluent” refers to a carrier, excipient, and diluent that does not irritate living organisms and does not inhibit the biological activity and properties of the administered compound. Examples of suitable carriers, excipients and diluents that may be included in such compositions include saline, sterile water, Ringer's solution, buffered saline, albumin injectable solution, glycerol, ethanol, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol. , starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, hypromellose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc. , magnesium stearate, and mineral oil. When pharmaceutically formulated, the composition may additionally contain conventional dispersants, fillers, extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, freeze-drying agents, etc. The exact proportions of these excipients may be determined by the solubility and chemical properties of the active compound, the chosen route of administration, or standard pharmaceutical practice.

The composition of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., and sterile injectable solutions according to conventional methods.

In the present invention, the pharmaceutical composition may be administered orally. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain the lecithin-like emulsifier plus at least one excipient, such as starch, calcium carbonate, or sucrose. It can be prepared by mixing sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate talc can also be used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives can be used. may be included. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous preparations, suspensions, emulsions, and freeze-dried preparations. Non-aqueous preparations and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.

In the present invention, the pharmaceutical composition may be administered in the form of an injection. Injectables may be formulated as intravenous or subcutaneous injections. In the case of parenteral injections, they may contain ingredients included in general injection compositions. For example, the injectable composition includes a liquid carrier such as sterile water, water for injection, or physiological saline. Additionally, it may further include amino acids, sugars, lipids, vitamins, electrolytes, pH adjusters, stabilizers, osmotic pressure adjusters, or solubilizers.

Various dosage forms, such as for injection and parenteral administration, can be prepared according to known or commonly used techniques in the art. Additionally, an effective amount of 4-hexylresorcinol may be administered by formulating a solution in saline or buffer solution immediately before administration in a form suitable for intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.

In addition, the present invention relates to the use of 4-hexylresorcinol or a pharmaceutically acceptable salt thereof for the production of a pharmaceutical composition for the prevention or treatment of male sexual dysfunction or male infertility due to diabetes, and a therapeutically effective amount of 4-hexylresorcinol. Provided is a method for preventing or treating male sexual dysfunction or male infertility due to diabetes, comprising administering rcinol or a pharmaceutically acceptable salt thereof to a subject in need thereof.

As used in the present invention, the term "prevention" refers to any action that suppresses or delays an immune disease by administering a pharmaceutical composition according to the present invention, and the term "treatment" refers to any action that suppresses or delays an immune disease by administering a pharmaceutical composition according to the present invention. It refers to all actions that improve or benefit the symptoms of an immune disease. In the present invention, “subject” refers to a subject in need of prevention or treatment of a disease, and more specifically, any mammal, such as humans and primates, as well as those in need of prevention or treatment of an immune disease. It includes livestock such as cows, pigs, sheep, horses, dogs, and cats without limitation, but is preferably a human.

In the above treatment method, the description of 4-hexylresorcinol, male sexual dysfunction, and male infertility can be applied as described above.

As used herein, “effective amount” means the amount necessary to delay or completely stop the onset or progression of a specific disease to be treated, and the effective amount of 4-hexylresorcinol contained in the pharmaceutical composition of the present invention is diabetes. This refers to the amount required to suppress testicular atrophy and increase the expression of LGR4 or testosterone in male sexual dysfunction and male infertility caused by .

The pharmaceutical composition of the present invention may be applied differently depending on the purpose of administration and disease. The amount of active ingredient actually administered depends on various related factors, such as the disease to be treated, the severity of the disease, whether or not it is co-administered with other drugs, the activity of the drug, sensitivity to the drug, the patient's age, gender, weight, food, and administration time. , administration route, and administration ratio of the composition may be appropriately selected by a person skilled in the art. The composition may be administered once a day or in divided doses 1-3 times, although the dosage and route of administration may be adjusted depending on the type and severity of the disease.

The pharmaceutical composition containing 4-hexylresorcinol of the present invention as an active ingredient can be administered at a dose of 0.01 mg/kg to 1 g/kg, more preferably 0.1 mg/kg to 100 mg/kg, It can be administered in a dosage of 1 mg/kg to 100 mg/kg, 10 mg/kg to 100 mg/kg, or 10 mg/kg to 50 mg/kg. Meanwhile, the dosage can be appropriately adjusted depending on the patient's age, gender, and condition.

The advantages and features of the present invention and methods for achieving them will become clear with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below and will be implemented in various different forms. The present embodiments are merely intended to ensure that the disclosure of the present invention is complete and that common knowledge in the technical field to which the present invention pertains is not limited. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims.

[Example]

Experimental Example 1: Confirmation of testicular atrophy inhibition ability of 4-hexylresorcinol (Figure 1)

The animal experiment was conducted with approval from Gangneung-Wonju National University IACUC (GWNU-2021-22, approval date: November 23, 2021). Twenty-two 4-week-old male rats with an average weight of 220 g were purchased and intravenously injected with streptozotocin (STZ) to induce type 1 diabetes. To confirm successful diabetes induction, fasting blood sugar was examined 3 days after STZ injection, and if blood sugar did not exceed 300 mg/dL, additional STZ was injected to induce diabetes in all 22 animals. Rats with induced diabetes were randomly divided into two groups. The experimental group was administered 12.8 mg/kg of 4-hexylresorcinol by subcutaneous injection into the skin tissue of the back twice a week, and the control group did not contain 4-hexylresorcinol. Only the injection solvent that was not used was injected in the same manner. 9 weeks after injection of 4-hexylresorcinol, the experiment was terminated, all animals were sacrificed, and testes were extracted and weighed. Separately from this experiment, in order to observe the effect of injection of 4-hexylresorcinol in normal rats in which diabetes was not induced by injection of STZ, testes were removed from rats injected with 4-hexylresorcinol in the same manner and weighed. were compared.

In diabetic rats, the testicular weight of the control group was 1.15 ± 0.51 g, and the testicular weight of the 4-hexylresorcinol group was 1.86 ± 0.26 g. The difference between the two groups was statistically significant (P=0.002). In healthy mice, the weight of the testis in the control group was 1.99 ± 0.17 g, and in the 4-hexylresorcinol group, the testis weight was 1.98 ± 0.19 g. There was no statistically significant difference between the two groups (P>0.05). Even when comparing the testicular weight of the control group of healthy rats and the testicular weight of the 4-hexylresorcinol group of diabetic rats, there was no statistically significant difference between the two groups (P>0.05). Through this, it was confirmed that testicular atrophy was suppressed more in rats injected with 4-hexylresorcinol than in male rats not injected with 4-hexylresorcinol.

Experimental Example 2: Confirmation of increased expression in testes following 4-hexylresorcinol treatment (Figure 2)

In experiments on diabetic rats, two testes were obtained from each experimental rat. One of them was used to extract protein from the tissue to investigate the expression level of LGR-4 through Western blot, and the other was used to fix it in formalin and embed it in paraffin.

To investigate the protein expression level of LGR-4 in tissues, the necessary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Proteins were extracted from tissues using an intratissue protein extraction kit. Considering the number of lanes that can be loaded at once, five animals from each group were randomly selected and the extracted proteins were mixed with sodium dodecyl sulfate buffer and separated by heating. The separated proteins were electrophoresed on a 10% polyacrylamide gel. The gel was transferred to a polyvinylidene difluoride membrane. After blocking, the membrane was probed with primary antibody (dilution ratio = 1:500). Blots were imaged and quantified using the ChemiDoc XRS system (Bio-Rad Laboratories, Hercules, CA, USA). The relative expression level was calculated as a ratio to the expression level of beta-actin. Differences in relative expression levels between the experimental and control groups were compared using independent samples' t-test.

The relative expression level of LGR4 in the control group was 0.47 ± 0.11, and in the 4-hexylresorcinol group it was 0.89 ± 0.26. When comparing the two groups, there was a statistically significant difference between the two groups (P=0.011). Through this, it was confirmed that when 4-hexylresorcinol was treated in male rats with induced diabetes, the expression of LGR-4 in the testes was higher than that in the control group.

Experimental Example 3: Confirmation of increased expression of LGR-4 and testosterone in testicular tissue following 4-hexylresorcinol treatment (Figure 3)

Samples embedded in paraffin were cut into 5 μm thick pieces and then hematoxylin and eosin staining and immunochemical staining were performed. Antibodies used in immunochemical staining were testosterone and LGR4. Testosterone is a representative male hormone synthesized in the testes, and after it is synthesized in the testes, it exists mostly in the form bound to protein, so antibodies were used to detect testosterone bound to protein. LGR4 is a marker of sperm production and is believed to be found in areas where sperm production is active.

Looking at the results of hematoxylin and eosin staining (HE stain), in the control group, a lot of sperm cell death was observed and almost no sperm tails were produced, so motility is expected to be reduced. However, in the group treated with 4-hexylresorcinol, high cell density and well-developed sperm tails could be observed.

In addition, looking at the results of immunochemical staining, it was observed that the expression of LGR4 and testosterone was much stronger in the group treated with 4-hexylresorcinol compared to the control group. Through this, it was confirmed that when male rats with induced diabetes were treated with 4-hexylresorcinol, sperm cell death was suppressed and the expression of LGR-4 and testosterone in the tissues increased.

Claims (10)

A pharmaceutical composition for preventing or treating male sexual dysfunction or male infertility due to diabetes, comprising 4-hexylresorcinol or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition of claim 1, wherein the male sexual dysfunction is erectile dysfunction, seborrhea, premature ejaculation, or retrograde ejaculation. The pharmaceutical composition of claim 1, wherein the male infertility is azoospermia, oligozoospermia, asthenospermia, or teratospermia. The pharmaceutical composition of claim 1, wherein the composition increases LGR-4 expression in testes. The pharmaceutical composition of claim 1, wherein the composition increases testosterone expression in the testes. The pharmaceutical composition according to claim 1, wherein the composition increases sperm production. The pharmaceutical composition of claim 1, wherein the composition inhibits testicular atrophy. The pharmaceutical composition of claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier. The pharmaceutical composition according to claim 1, wherein the composition is administered at a dosage of 0.1 mg/kg to 100 mg/kg. The pharmaceutical composition of claim 1, wherein the composition is administered orally or parenterally.