Use of inosine in the preparation of a medicament for promoting testosterone secretion

Abstract

This invention discloses the application of inosine in the preparation of drugs that promote testosterone secretion, belonging to the field of biomedical technology. The effective active ingredient of the above-mentioned drugs is only inosine or its pharmaceutically acceptable salts. The dosage forms of the above-mentioned drugs are tablets, capsules, oral liquids, lozenges, granules, powders, pills, powders, ointments, elixirs, suspensions, powders, or injections. The routes of administration of the above-mentioned drugs are oral, transdermal, intramuscular, subcutaneous, or intravenous injection. Since inosine exists under physiological conditions and can be metabolized by intestinal microorganisms, reasonable supplementation of inosine is a safer and more effective method for restoring testosterone. Inosine helps maintain the homeostasis of the intestinal flora, and the experimental results of this invention further show that inosine can restore the damaged intestinal barrier, thus making it safer. Moreover, this invention does not directly supplement exogenous testosterone, therefore it is less likely to induce spermatogenesis disorders or promote prostatic hyperplasia.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more specifically to the application of inosine in the preparation of drugs that promote testosterone secretion. Background Technology

[0002] Sex hormones have a significant impact on an individual's physiological functions, development, behavior, and emotions. Therefore, a decline in testosterone levels may lead to adverse changes such as developmental delays, sexual and reproductive dysfunction, autonomic nervous system disorders, and mental and psychological problems, seriously affecting men's physical and mental health and quality of life.

[0003] Currently, testosterone replacement therapy (TST) is used clinically to treat insufficient testosterone secretion caused by hypogonadism. It involves exogenously supplementing testosterone to achieve normal physiological concentrations in the body, thereby eliminating the physiological changes and clinical symptoms caused by partial testosterone deficiency. While this treatment method is direct and highly effective, its safety is significantly compromised, and it is prone to a series of side effects, such as impaired spermatogenesis, gastrointestinal dysfunction, increased red blood cell and hemoglobin levels, significant liver toxicity, promotion of acute urinary retention, and even induction of prostate cancer.

[0004] Specifically, excessive use of TST can easily lead to spermatogenesis disorders. Under normal circumstances, the hypothalamus-pituitary-testis axis regulates testosterone levels in men. Exogenous testosterone not only stimulates prostate hyperplasia but also causes negative feedback inhibition of the hypothalamus-pituitary-testis axis, affecting luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels related to spermatogenesis, thereby inhibiting spermatogenesis. Of course, excessive use of TST can also stimulate non-cancerous growth of the prostate (benign prostatic hyperplasia) and the growth of existing prostate cancer; it can also lead to gastrointestinal dysfunction and intestinal flora imbalance.

[0005] To avoid the aforementioned problems, testosterone patches can be used. These are testosterone replacement therapy patches for men with hypogonadism, applied to the skin on the back, abdomen, upper arms, or groin. However, this testosterone replacement therapy still has many adverse reactions, such as local skin irritation, rashes, pustular eruptions, testicular pain, night sweats, lower limb pain, hearing loss, and upper respiratory tract infections. Therefore, there is still a need to find safer testosterone-boosting drugs. Summary of the Invention

[0006] Based on the above-mentioned technical problems, this invention proposes the application of inosine in the preparation of drugs that promote testosterone secretion.

[0007] The technical solution adopted in this invention is:

[0008] The application of inosine in the preparation of drugs that promote testosterone secretion.

[0009] The only active ingredient in the above-mentioned drugs is inosine or its pharmaceutically acceptable salt.

[0010] The dosage forms of the above-mentioned drugs are tablets, capsules, oral liquids, lozenges, granules, powders, pills, powders, ointments, elixirs, suspensions, powders, or injections.

[0011] The above-mentioned drugs can be administered orally, transdermally, intramuscularly, subcutaneously, or intravenously.

[0012] Furthermore, drugs containing the active ingredient inosine can restore testosterone levels on the basis of taking the narrow-spectrum antibiotic colistin.

[0013] The beneficial technical effects and principles of this invention are as follows:

[0014] Sex hormones are metabolized through the enterohepatic circulation pathway, therefore testosterone is often influenced by the gut microenvironment. Gut microorganisms, including bacteria and pathogens, shape the gut microenvironment and also affect local immune function. The intestinal mucus layer, the first line of defense in the intestinal barrier, separates bacteria and pathogens from intestinal mucosal epithelial cells, playing a crucial role in human health. Together, they constitute the gut microenvironment. When factors such as sex hormones, medications, age, diet, and emotions cause gut microbiota imbalance, large amounts of lipopolysaccharide (LPS) accumulate, inducing intestinal mucosal damage, inflammation, and impairing testicular function. Therefore, abnormal changes in the gut microenvironment not only harm gut health but also affect testosterone biosynthesis.

[0015] The inventors have discovered through years of research that the method of restoring normal testosterone levels by regulating gut health is safer and more reliable.

[0016] This invention utilizes inosine to prepare a drug that promotes testosterone secretion. While protecting the intestinal mucus layer, it effectively promotes testosterone secretion and restores testosterone levels. The drug prepared by this invention also has advantages such as being safer, more effective, and easier to use, having no obvious side effects, and being easily accepted by patients, thus meeting the needs of normal men to restore testosterone levels.

[0017] In summary, firstly, based on the discovery that the narrow-spectrum antibiotic colistin causes intestinal flora imbalance and inhibits testosterone in normal male mice, this invention, by administering inosine (a metabolite of the intestinal flora that can be synthesized by specific intestinal flora), can significantly restore the testosterone imbalance in normal male mice caused by the narrow-spectrum antibiotic colistin. Secondly, the inosine component of this invention can effectively improve the damage to the intestinal mucus layer caused by the narrow-spectrum antibiotic colistin, reduce the increase in LPS caused by the narrow-spectrum antibiotic colistin, and avoid the damage of the narrow-spectrum antibiotic colistin to the intestinal barrier and the body. Finally, the drug containing inosine used in this invention has the advantages of being easily obtained, convenient to take, protecting the intestinal mucus layer, and regulating the inflammatory response. Attached Figure Description

[0018] Figure 1 This study investigated the changes in gut microbiota caused by the narrow-spectrum antibiotic colistin. Two mouse groups, CTR and COL, were included. Figure a shows the α-diversity of the mouse gut microbiota, where Observed, Shannon, Chao1, Simpson, and ACE all reflect α-diversity. Figure b is a box plot of the absolute abundance of gut microbiota in mice. Figure c is principal component analysis (PCA) based on the absolute abundance of ASV. Figure d shows the relative β-diversity analysis of the microbiota.

[0019] Figure 2 To investigate the use of inosine to restore the damaged intestinal barrier, four groups of mice were set up: CTR, COL, INO, and CI. In the figure, a is a microscopic image of Alcian blue staining of the mouse colon; b is a statistical graph of the thickness of the intestinal mucus layer in a; and c is the concentration of LPS in the mouse serum.

[0020] Figure 3 Inosine was used to restore abnormal serum testosterone secretion in male mice caused by intestinal barrier damage. Detailed Implementation

[0021] This invention provides the application of inosine in the preparation of drugs that promote testosterone secretion, mainly by protecting the intestinal mucus layer and regulating the intestinal microenvironment to meet the needs of normal men to restore testosterone. When damage to the intestinal mucus layer occurs, inosine can promote testosterone secretion by restoring the damaged barrier.

[0022] Inosine helps maintain gut microbiota homeostasis, and the experimental results of this invention further demonstrate that inosine can restore the damaged intestinal barrier, thus making it safer. Moreover, this invention does not directly supplement exogenous testosterone, therefore it is less likely to induce spermatogenesis disorders or promote benign prostatic hyperplasia.

[0023] Since inosine exists under physiological conditions and can be metabolized by gut microbes (although inosine can be metabolized by gut microbes, it still needs to be directly supplemented from the outside to restore testosterone levels), reasonable inosine supplementation is a safer and more effective way to restore testosterone.

[0024] The only active ingredient in the above-mentioned drugs is inosine or its pharmaceutically acceptable salt.

[0025] Furthermore, the combination of the narrow-spectrum antibiotic colistin and inosine is preferred; inosine is more effective than colistin alone. Specifically, inosine can restore testosterone levels in mice on the basis of oral administration of the narrow-spectrum antibiotic colistin.

[0026] The dosage forms of the aforementioned drugs may include tablets, capsules, oral liquids, lozenges, granules, powders, pills, powders, ointments, elixirs, suspensions, or injections. These dosage forms, such as powders, offer advantages such as easier storage and transportation, simpler operation and use, and higher patient acceptance.

[0027] The above-mentioned drugs can be administered orally, transdermally, intramuscularly, subcutaneously, or intravenously.

[0028] The term "pharmaceutically acceptable" as used above refers to a substance suitable for human use without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a substance with a reasonable benefit / risk ratio. Inosine, mentioned above, is an active substance that can be used in combination with carriers or excipients to prepare a drug. Carriers or excipients include one or more of the following: binders, fillers, diluents, tableting agents, lubricants, disintegrants, colorants, flavoring agents, and humectants.

[0029] The new applications provided by the present invention will be further illustrated below through specific embodiments.

[0030] Example 1

[0031] Analysis of the gut microbiota in mice:

[0032] (1) Grouping, feeding and administration of male mice: C57BL / 6N male mice (7 weeks old and above sexually mature) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. All mice were maintained under specific pathogen-free conditions. After acclimatization for 1 week, they were divided into two groups of 6 mice each and treated with drugs for 7 days.

[0033] Normal group (CTR): Administered sterile drinking water.

[0034] Colistin treatment group (COL): 1 mg / mL Colistin (Aladdin, C114323) was added to sterile drinking water.

[0035] (2) Analysis of the abundance and composition of gut microbiota: Fresh feces of mice were collected on the 7th day of the experiment, stored at -80℃, and the composition of gut microbiota was analyzed by 16s absolute quantitative sequencing. At the end of the experiment, the mice were sacrificed, and aseptic operation was performed when collecting feces from the cecum segment. The feces were stored at -80℃ and the changes in gut microbiota were analyzed.

[0036] (3) Results:

[0037] 16S absolute quantitative sequencing analysis: such as Figure 1 As shown, the inventors collected fecal samples after 7 days of colistin treatment and performed accurate 16S absolute quantitative sequencing. Result a indicates consistency with its narrow antibacterial spectrum, and ns indicates no significant difference in α-diversity between the colistin treatment group and the control group. α-diversity and β-diversity reflect biodiversity at two different spatial scales. Indices reflecting α-diversity include: Observed, Shannon index, Chao1, Simpson index, and ACE. Observed: indicates the number of species contained in the sample; a higher Shannon index indicates higher biodiversity; the Simpson index represents species diversity; higher species diversity means a lower probability that two samples belong to the same species and a higher probability that they belong to different species; Chao1: represents an index that estimates the number of OTUs contained in the sample; ACE index: assesses the richness and evenness of species composition in the sample. Result b shows that the absolute abundance of total bacterial load per gram of fecal sample did not differ significantly between the two groups. Results c show that principal component analysis (PCA) based on the absolute abundance of ASV revealed significant changes in the gut microbiota structure, but the differences were not statistically significant (ANOSIM, P = 0.194; Adonis, P = 0.067). Results d show that relative β-diversity analysis revealed a clear separation between the colistin treatment group and the control group. Significant differences in gut microbiota structure were observed among the groups (ANOSIM, P = 0.031; Adonis, P = 0.03). In conclusion, these results indicate that Colistin significantly affects the gut microbiota of mice.

[0038] Example 2

[0039] The protective effect of inosine on the intestinal barrier in mice:

[0040] (1) Grouping, feeding and administration of male mice: Animal conditions were the same as in Example 1, only the grouping was different;

[0041] Normal group (CTR): each animal was given sterile drinking water and 0.1 ml of sterile phosphate-buffered saline (PBS) by gavage daily.

[0042] Colistin treatment group (COL): 1 mg / mL Colistin (Aladdin, C114323) was added to sterile drinking water, and each animal was given 0.1 ml of sterile phosphate-buffered saline (PBS) by gavage daily.

[0043] Inosine treatment group (INO): each animal was given sterile drinking water and 0.1 ml of sterile phosphate-buffered saline (PBS) containing 300 mg / kg Inosine by gavage daily.

[0044] Colistin combined with Inosine treatment group (CI): 1 mg / mL Colistin (Aladdin, C114323) was added to sterile drinking water, and each animal was given 0.1 ml of sterile phosphate-buffered saline (PBS) containing 300 mg / kg Inosine by gavage daily.

[0045] (2) Method for detecting colonic mucus layer: After sacrifice of mice, colons were collected, fixed with Carnoy fixative, embedded in paraffin, and sectioned at 5 μM. Paraffin sections were dewaxed, dehydrated, and stained with alcine blue (Servicebio, G1049). The thickness of colonic mucus layer was measured in 10 random fields of view on each section using ImageJ software (NIH, USA).

[0046] (3) Detection method of serum LPS: After administering an anesthetic to mice via intraperitoneal injection, serum samples were collected, stored at 4°C, and the concentration of LPS (Cloud-Clone Crop, HEB526Ge) was detected using an ELISA kit according to the manufacturer's instructions.

[0047] (4) Results:

[0048] Figure 2 Figure a shows the results of Alcian blue staining. Intestinal mucus is continuously distributed on the surface of the intestinal mucosa and in contact with the intestinal contents, playing a crucial role in maintaining normal intestinal function. Figure b is a statistical graph of intestinal mucus layer thickness. Figure c is a statistical graph of LPS concentration in mouse serum. * indicates statistically significant differences, ns indicates no statistically significant differences. Colistin causes thinning of the intestinal mucus layer in mice. Figure 2 (a and b) significantly increased serum LPS levels ( Figure 2 In the middle (c), the combination of Colistin and Inosine (CI group) can reduce the elevation of serum LPS and restore the thickness of the intestinal mucus layer. Figure 2(Ac). This demonstrates that Inosine can restore the damaged intestinal mucus layer and has a protective effect on the intestinal barrier.

[0049] Example 3

[0050] The restorative effect of inosine on testosterone levels in mice:

[0051] (1) Grouping, feeding and administration of male mice: Same as in Example 2.

[0052] (2) Method for detecting serum testosterone: After administering an anesthetic to mice via intraperitoneal injection, serum samples were collected, stored at 4°C, and the concentration of LPS (Cloud-Clone Crop, HEB526Ge) was detected using an ELISA kit according to the manufacturer's instructions.

[0053] (3) Results:

[0054] Figure 3 This is a statistical graph showing the concentration of testosterone in mouse serum. * indicates a statistically significant difference, and ns indicates no statistically significant difference. Compared to the control group, Colistin significantly reduced testosterone secretion in mice, while the combination of Colistin and Inosine (CI group) promoted the recovery of testosterone to normal levels (i.e., no significant difference in testosterone levels compared to the control group). This indicates that Inosine can restore testosterone secretion by protecting the intestinal mucus layer.

[0055] In summary, Colistin leads to thinning of the intestinal mucus layer and a significant increase in serum LPS, thereby inhibiting testosterone levels in normal male mice. Administration of Inosine antagonizes the adverse effects of Colistin, restores the intestinal mucus layer, and ultimately restores testosterone levels.

Claims

1. The use of inosine or a pharmaceutically acceptable salt thereof as the sole active ingredient in the preparation of drugs that promote testosterone secretion.

2. The application according to claim 1, characterized in that, The dosage form of the drug is tablets, capsules, oral liquid, lozenges, granules, powders, suspensions, or injections.

3. The application according to claim 1, characterized in that, The drug can be administered orally, intramuscularly, subcutaneously, or intravenously.

4. The application according to claim 1, characterized in that, The drug can restore testosterone levels when taken on the basis of the narrow-spectrum antibiotic colistin.

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