Application of urolithin A in preparation of product for improving endogenous estrogen level

Abstract

The invention relates to an application of urolithin A in preparation of a product for improving an endogenous estrogen level. The molecular formula of the urolithin A is C13H8O4, and the structural formula of the urolithin A is shown as a formula (I). The improvement of the endogenous estrogen level comprises the following steps: selectively regulating and controlling synthesis pathways of adrenal hormone and ovarian estrogen, and synergistically improving the in-vivo estrogen level. Besides, the urolithin A has good hormone regulation selectivity, can effectively inhibit cortisol secretion, and has no obvious influence on aldosterone level, so that blood pressure regulation and body fluid homeostasis are prevented from being interfered. The action characteristics lay a foundation for the application of the urolithin A in improvement of female climacteric hormone disorder and related health support directions, and the urolithin A has a wide prospect as an endogenous hormone regulation strategy. (I)

Description

Technical Field

[0001] This article relates to the field of biomedicine, specifically the use of urolithin A in the preparation of products that increase endogenous estrogen levels and improve menopausal hair loss caused by declining estrogen levels in women. Background Technology

[0002] During menopause, women often experience a significant drop in estrogen levels, leading to various symptoms such as menopausal hair loss, hot flashes, insomnia, mood swings, and osteoporosis. While hormone replacement therapy can alleviate these symptoms, safety concerns limit its long-term use (Li et al., Med, 2023). Therefore, developing safer small-molecule interventions that can promote the body's own estrogen secretion has significant clinical value.

[0003] Metenidone chemical structure Metyrapone promotes the secretion of dehydroepiandrosterone (DHEA) and sex hormones, and inhibits cortisol production. However, it also inhibits aldosterone production, which can affect blood pressure. Therefore, it is not suitable for improving menopausal estrogen deficiency in women. The fundamental reason is that metyrapone works by inhibiting 11-β-hydroxylase (CYP11B1), an enzyme that plays a crucial role in the synthesis of cortisol and aldosterone. Therefore, this inhibition inevitably blocks the synthesis of aldosterone.

[0004] Current estrogen regulation methods generally lack hormone selectivity, easily causing non-specific interference with non-target hormones such as aldosterone, which may lead to side effects or metabolic disorders. Therefore, there is an urgent need to develop a regulatory strategy that can specifically regulate the synthesis of functional hormones while avoiding adverse effects on other key hormones. Summary of the Invention

[0005] The following is an overview of the subject matter described in detail herein. This overview is not intended to limit the scope of protection of this application.

[0006] The urolithin A used in this application is a natural active ingredient derived from pomegranate. It can promote estrogen levels and reduce cortisol levels by regulating endogenous hormone synthesis pathways, while having no effect on aldosterone levels, demonstrating excellent hormone regulation properties. These properties indicate its significant potential for intervention in hormonal imbalances related to menopause in women, and it is expected to be used to support hormonal balance regulation during menopause, alleviating physiological discomfort caused by declining estrogen levels, such as hair loss, hot flashes, mood swings, sleep disorders, and metabolic disturbances.

[0007] The first aspect of this application provides the use of urolithin A in the preparation of products for increasing endogenous estrogen levels to improve menopausal hair loss caused by estrogen decline in women during menopause.

[0008] In one exemplary embodiment, the molecular formula of urolithin A is C2. 13 H8O4, the structural formula is shown in formula (I): (I).

[0009] In one exemplary embodiment, increasing endogenous estrogen levels includes: selectively regulating the synthesis of adrenal hormones and ovarian estrogen to synergistically increase estrogen levels in the body; optionally, it also includes: inhibiting cortisol secretion, and / or having no significant effect on aldosterone secretion.

[0010] In one exemplary embodiment, increasing endogenous estrogen levels includes: promoting the synthesis of dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), estrone, pregnenolone, and estradiol in adrenal cells, promoting estrogen synthesis in ovarian granulosa cells, and further enhancing its effect of increasing overall estrogen levels; it also includes: inhibiting cortisol secretion, and having no significant effect on aldosterone secretion.

[0011] In one exemplary embodiment, the product is applied to a mammal; preferably, to a human.

[0012] In one exemplary embodiment, the product is a medicine, daily chemical product, health product, or food.

[0013] In one exemplary embodiment, the drug can be used to support, regulate, or improve physiological conditions associated with decreased estrogen levels during menopause in women, while maintaining stable blood pressure and fluid balance.

[0014] In one exemplary embodiment, the physiological state associated with the decline in estrogen levels during female menopause includes, but is not limited to: Menopausal-related hormonal imbalances (such as decreased estrogen, decreased DHEA, etc.); Menopausal symptoms (such as hot flashes, night sweats, mood swings, anxiety, and sleep disturbances); Metabolic syndrome or menopausal metabolic disorders; Menopausal estrogen decline can lead to gray hair, hair loss, muscle loss, increased body fat percentage, decreased endurance, osteoporosis, and other related symptoms.

[0015] In one exemplary embodiment, the dosage form of the drug includes any one of the following: cream, patch, ointment, cream preparation, gel, spray, tablet, granule, capsule, oral liquid preparation, pill, suspension, drop pill, and injection.

[0016] In one exemplary embodiment, when the drug is used on a human body, the dosage of urolithin A is 200-1000 mg / day; for example, 200 mg / day, 300 mg / day, 400 mg / day, 500 mg / day, 600 mg / day, 700 mg / day, 800 mg / day, 900 mg / day, 1000 mg / day, or any two of these values ​​as a range between endpoints, but not limited to the listed values, other unlisted values ​​within this range are also applicable; Preferably, when the drug is used on a human body, the dosage of urolithin A is 250-1000 mg / day; more preferably, the dosage of urolithin A is 250 mg / day.

[0017] In one exemplary embodiment, the daily chemical product is a cosmetic, care product, or beauty product.

[0018] In one exemplary embodiment, the product uses urolithiasis A as the sole active ingredient.

[0019] In one exemplary embodiment, the product may also include other active ingredients.

[0020] In one exemplary embodiment, the product may further include pharmaceutically, biologically, or cosmetically acceptable carriers or excipients.

[0021] In one exemplary embodiment, the product may be applied by any of the following methods: topical application to the skin, oral administration, or injection.

[0022] The second aspect of this application provides a drug for increasing endogenous estrogen levels, said drug having urolithin A as the sole active ingredient.

[0023] The third aspect of this application provides the use of urolithin A in the preparation of a medicament that supports, regulates or improves physiological conditions associated with decreased estrogen levels during menopause in women and maintains stable blood pressure and fluid balance.

[0024] In one exemplary embodiment, the drug has urolithiasis A as the sole active ingredient; optionally, it may also include other active ingredients.

[0025] In one exemplary embodiment, the drug is administered to a mammal; preferably, the drug is administered to a human.

[0026] In one exemplary embodiment, the medicament may further include a pharmaceutically or biologically acceptable carrier or excipient.

[0027] In one exemplary embodiment, the drug may be administered by any of the following methods: topical application to the skin, oral administration, or injection.

[0028] A fourth aspect of this application provides a method for increasing endogenous estrogen levels, the method comprising administering the aforementioned drug or product containing urolithin A to a patient with appropriate need. Compared with existing related technologies, this application has the following technical advantages: This application discloses the multiple effects of urolithin A in regulating endogenous hormone secretion, particularly its effect on increasing estrogen levels. Specifically, urolithin A can significantly promote the synthesis and secretion of dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), estrone, pregnenolone, and estradiol in adrenal cells, and promote the production of ovarian estradiol in ovarian granulosa cells, thus synergistically increasing estrogen levels. Furthermore, urolithin A exhibits good selective hormone regulation, effectively inhibiting cortisol secretion without significantly affecting aldosterone levels. Based on this, urolithin A shows significant potential in intervening in hormonal imbalances related to menopause in women, and can be used to support hormonal balance regulation during menopause, alleviating physiological discomfort caused by declining estrogen levels, such as hair loss, hot flashes, mood swings, sleep disorders, and metabolic disturbances. It is worth emphasizing that urolithin A does not affect aldosterone secretion, allowing it to regulate functional hormone levels without interfering with blood pressure regulation mechanisms, thus improving its safety and specificity. Animal experiments conducted in this application showed that oral administration of urolithin A effectively increased serum estradiol levels in middle-aged female mice, significantly reduced cortisol levels, and had no significant effect on aldosterone. Visual observation revealed that mice treated with urolithin A had denser, darker, and shinier hair, significantly improved hair loss, and increased hair growth rate, suggesting its potential value in improving female hair loss caused by decreased estrogen levels during menopause.

[0029] Other features and advantages of this application will be set forth in the following description, and will be apparent in part from the description, or may be learned by practicing the application. Other advantages of this application can be realized and obtained by means of the embodiments described in the description and the accompanying drawings. Attached Figure Description

[0030] The accompanying drawings are used to provide an understanding of the technical solutions of this application and constitute a part of the specification. They are used together with the embodiments of this application to explain the technical solutions of this application and do not constitute a limitation on the technical solutions of this application.

[0031] Figure 1 Results of hormone secretion levels in H295R cell lines treated with urolithin A; Figure 2 Detection of mRNA levels of hormone secretion-related enzymes in H295R cell lines treated with urolithin A; Figure 3 Results of estradiol secretion levels in KGN cell lines treated with urolithin A; Figure 4 The animal appearance is compared between female mice treated with urolithin A by gavage and female mice in the control group in Example 4. Figure 5 This is a comparison of the results of the gavage treatment of female mice with urolithin A and the control group female mice after hair removal in Example 4. Figure 6 This is a comparison of the serum estradiol concentrations in female mice treated with urolithin A via gavage and in the control group in Example 4. Figure 7 This is a comparison of serum cortisol concentrations in female mice treated with urolithin A via gavage and in the control group in Example 4. Figure 8 This is a comparison of serum aldosterone concentrations in female mice treated with urolithin A via gavage and in the control group in Example 4. Detailed Implementation

[0032] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] In this application, urolithin A achieves its biological effects by specifically regulating the steroid synthesis pathway in adrenal cells and ovarian granulosa cells. On the one hand, urolithin A can upregulate the expression of key enzymes involved in the synthesis of various sex hormones, enhancing the biosynthesis of hormones such as dehydroepiandrosterone (DHEA), DHEA sulfate, estrone, estradiol, and pregnenolone. On the other hand, it has a selective inhibitory effect on cortisol synthase, thereby reducing cortisol secretion levels and alleviating the physiological burden caused by its excess. Crucially, urolithin A has no significant effect on the aldosterone synthesis pathway, demonstrating excellent selective hormone regulation. This characteristic overcomes the common drawback of existing technologies that "promote one type of hormone while interfering with other hormones," providing a new solution for achieving precise and balanced regulation of adrenal hormone and estrogen secretion.

[0034] The raw materials used in this application are all conventional products on the market.

[0035] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0036] Experimental methods not specified in the examples are generally performed under standard conditions or as recommended by the manufacturer.

[0037] Example 1. Detection of hormone secretion levels in H295R cell lines treated with urolithin A 1. Materials and solvents: Urolithiasis A was purchased from GLPBIO, product number GC15168; Cell line treated: H295R adrenocortical carcinoma cell line; Treatment concentration: 50 μM; Solvent: DMSO.

[0038] 2. Processing time: 48 hours.

[0039] 3. Experimental steps: The H295R cell line treated with 50 μM urolithin A for 48 hours was used as the experimental group, and the H295R cell line treated with an equal volume of DMSO for 48 hours was used as the control group. Simultaneously, a blank group (DMEM / F12 medium without added cells) was set up to subtract the hormone levels at the culture baseline.

[0040] After the experiment, the hormone secretion levels in the culture medium of H295R cells in the control and experimental groups were measured. The results are shown below. Figure 1 .

[0041] Results and conclusions: Figure 1 As shown, compared with the control group, after treatment with 50 μM urolithin A, the H295R cell line in the experimental group showed an increase of approximately 434% in dehydroepiandrosterone (DHEA), approximately 171% in DHEA sulfate, approximately 1598% in estrone, approximately 180% in pregnenolone, and approximately 80% in estradiol. Simultaneously, it reduced cortisol levels to 67% of the control group, with no significant effect on aldosterone secretion. Based on these results, urolithin A can significantly promote the secretion of DHEA, DHEAS, estrone, pregnenolone, and estradiol from adrenal cells, while effectively inhibiting cortisol secretion, and has no significant effect on aldosterone secretion. This fully demonstrates the technical advantage of urolithin A in precisely regulating adrenal hormone secretion.

[0042] Example 2. Detection of mRNA levels of hormone secretion-related enzymes in H295R cell lines treated with urolithin A Cells from each group in Example 1 were collected, lysed, and total RNA was extracted. After reverse transcription into cDNA, the mRNA expression levels of hormone synthesis-related enzymes in each group were detected using real-time quantitative PCR (qPCR). The results are shown in [Figure 1]. Figure 2 .

[0043] Conclusion: Figure 2 As shown, compared with the control group, treatment with 50 μM urolithin A significantly increased the mRNA levels of SULT2A1 and CYP19A1 enzymes in cells. These two enzymes play a crucial role in the synthesis of dehydroepiandrosterone sulfate (DHEA), estrone, and estradiol, consistent with the increased secretion of DHEA, estrone, and estradiol in Example 1. Simultaneously, treatment with 50 μM urolithin A decreased the mRNA level of CYP11B1 enzyme in cells. This enzyme plays an important role in cortisol synthesis, consistent with the decreased cortisol secretion observed in Example 1. Furthermore, 50 μM urolithin A treatment had no significant effect on the mRNA level of CYP11B2 enzyme in cells. CYP11B2 is the rate-limiting enzyme in aldosterone synthesis, consistent with the unchanged aldosterone secretion level observed in Example 1.

[0044] Example 3. Results of estradiol secretion level detection in the ovarian granulosa cell line KGN treated with urolithin A 1. Materials and solvents: Urolithiasis A was purchased from GLPBIO, product number GC15168; Cell line treated: KGN ovarian granulosa cell line; Treatment concentration: 10 μM; Solvent: DMSO.

[0045] 2. Processing time: 48 hours.

[0046] 3. Experimental steps: The experimental group consisted of KGN cells treated with 10 μM urolithin A for 48 hours, while the control group consisted of KGN cells treated with an equal volume of DMSO for 48 hours. Additionally, a blank group (DMEM / F12 medium without added cells) was used to subtract the baseline hormone levels in the culture medium.

[0047] After the experiment, the hormone secretion levels in the KGN cell culture medium of the experimental and control groups were measured, and the results are shown below. Figure 3 .

[0048] Results and conclusions: Figure 3As shown, compared with the control group, the KGN cell line in the experimental group showed a significant increase in estradiol secretion after treatment with 10 μM urolithin A, increasing by approximately 77%. This indicates that urolithin A can significantly promote estrogen secretion by ovarian granulosa cells.

[0049] Example 4. Results of gavage treatment of female mice with urolithin A Experimental Methods: Eleven-month-old SPF-grade C57BL / 6J middle-aged female mice with noticeably white and sparse fur were selected. Mice were randomly divided into an experimental group (n=10 per group) and a control group (n=10 per group). The experimental group received urolithin A via gavage at a dose of 50 mg / kg / day (equivalent to 250 mg / day for a 60 kg adult), using 0.5% sodium carboxymethyl cellulose as the dissolving carrier. The control group received the same dose of 0.5% sodium carboxymethyl cellulose via gavage. The treatment period was 5 months.

[0050] During the experiment, the appearance of mice in each group was observed and the serum hormone levels of the mice were measured. The experimental results are as follows: Figures 4 to 8 As shown: The appearance of the experimental animals in each group was observed after 2 months of experimentation, and the results are as follows: Figure 4 As shown in the figure. The results showed that, compared with the control group (Ctrl), the experimental group (UA) mice treated with urolithin A had thicker, darker fur and significantly less hair loss; In the third month of the experiment, the skin on the backs of the mice was shaved, and hair regrowth was observed two months later. Figure 5 As shown in the figure, the results indicated that, compared with the control group (Ctrl), the experimental group mice treated with urolithin A (UA) had significantly denser hair regeneration in the shaved area, suggesting that urolithin A treatment can significantly promote hair growth. Serum hormone levels in each group of experimental animals were measured at 3 months of the experiment, and the results are shown below. Figures 6 to 8 As shown. Figure 6 The serum estradiol level in mice treated with urolithin A was 1.5 times that of the control group, indicating that urolithin A can significantly increase estrogen levels. Figure 7 The serum cortisol level in mice treated with urolithin A decreased to 80% of that in the control group, indicating that urolithin A has a significant inhibitory effect on cortisol. Figure 8 The serum aldosterone levels in the experimental group of mice treated with urolithin A were not significantly different from those in the control group, proving that urolithin A does not interfere with aldosterone secretion.

[0051] This experiment showed that feeding urolithin A could increase estradiol secretion by about 53%, reduce cortisol levels to 80% of the control group, and have no effect on aldosterone levels.

[0052] In summary, the small molecule urolithin A used in this application significantly increases estradiol secretion by specifically regulating the synthesis pathways of adrenal and ovarian hormones, while effectively reducing cortisol levels and having no effect on aldosterone secretion. This solves the problem of poor selectivity and easy hormonal imbalance in existing technologies in hormone regulation, and provides a safe and efficient solution for alleviating common physiological and psychological symptoms (such as hair loss, hot flashes, mood swings, metabolic disorders, and sleep disturbances) caused by hormonal changes during menopause in women.

[0053] Although embodiments of this application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting this application. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of this application.

[0054] The following references are incorporated in this paper as part of this paper: [References] 1. Daniel, E., Aylwin, S., Mustafa, O., Ball, S., Munir, A.,Boelaert, K., ... & Newell-Price, J. (2015). Effectiveness of metyrapone intreating Cushing's syndrome: a retrospective multicenter study in 195patients. The Journal of Clinical Endocrinology & Metabolism, 100(11), 4146-4154. 2. Sigalas, PD, Garg, H., Watson, S., McAllister-Williams, RH, & Ferrier, IN (2012). Metyrapone in treatment-resistant depression. Therapeutic advances in psychopharmacology, 2(4), 139-149.

Claims

1. The use of urolithin A in the preparation of products for increasing endogenous estrogen levels, wherein, The molecular formula of urolithin A is C 13 H8O4, the structural formula is shown in formula (I): （I）。 2. The use according to claim 1, wherein, Increasing endogenous estrogen levels includes selectively regulating the synthesis of adrenal hormones and ovarian estrogen to synergistically increase estrogen levels in the body; optionally, it also includes inhibiting cortisol secretion and / or having no significant effect on aldosterone secretion.

3. The use according to claim 2, wherein, Increasing endogenous estrogen levels includes: promoting the synthesis of dehydroepiandrosterone, dehydroepiandrosterone sulfate, estrone, pregnenolone, and estradiol in adrenal cells, and promoting the production of ovarian estradiol in ovarian granulosa cells; it also includes: inhibiting cortisol secretion, and has no significant effect on aldosterone secretion.

4. The use according to any one of claims 1 to 3, wherein, The product is applied to mammals; optionally, the product is applied to humans.

5. The use according to any one of claims 1 to 3, wherein, The products mentioned are pharmaceuticals, daily chemical products, health products, or food.

6. The use according to claim 5, wherein, The dosage form of the drug includes any one of the following: cream, patch, ointment, cream preparation, gel, spray, tablet, granule, capsule, oral liquid preparation, pill, suspension, drop pill, injection; and / or When the drug is used on humans, the dosage of urolithiasis A is 200-1000 mg / day.

7. The use according to claim 5, wherein, The daily chemical products mentioned are cosmetics, care products, or beauty products.

8. The use according to claim 5, wherein, The product uses urolithiasis A as its sole active ingredient. Optionally, the product may also include other active ingredients; Optionally, the product may also include pharmaceutically, biologically, or cosmetically acceptable carriers or excipients.

9. The use according to any one of claims 1 to 3, wherein, The product is applied by any of the following methods: topical application to the skin, oral administration, or injection.

10. The use of urolithin A in the preparation of drugs to support, regulate, or improve physiological states associated with declining estrogen levels during menopause in women and to maintain stable blood pressure and fluid balance; among which, The drug has urolithiasis A as its sole active ingredient; optionally, it may also include other active ingredients.

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