Method and composition for protecting or maintaining cardiovascular health

Abstract

A composition and method for protecting or maintaining cardiovascular health, improving sexual satisfaction or sexual health or sexual ability, or relieving sexual dysfunction. The composition comprises salidroside or a pharmaceutically acceptable salt, ester or derivative thereof; and ergothioneine or a pharmaceutically acceptable salt, acid, ester, analogue or derivative thereof.

Description

Methods and compositions for protecting or maintaining cardiovascular health Technical Field

[0001] This invention belongs to the field of pharmaceutical and health care or nutritional supplement technology; specifically, it relates to compositions and methods for protecting or maintaining cardiovascular health, improving sexual satisfaction or sexual health, or improving sexual dysfunction. Background Technology

[0002] Cardiovascular health is one of the major public health challenges worldwide, causing millions of deaths annually, with coronary heart disease and stroke being the most common causes. While the incidence and mortality rates of cardiovascular disease have declined in some developed countries through lifestyle improvements and medical interventions, they continue to rise in many developing countries due to population aging and the prevalence of unhealthy lifestyles. In China, the prevalence of cardiovascular disease continues to rise, with estimated numbers reaching hundreds of millions, and the increase is particularly pronounced in rural areas. An increasing number of people are taking dietary supplements to prevent and improve their cardiovascular health, aiming to clear blood vessels, lower blood pressure, cleanse and quickly eliminate toxins from blood vessels, and improve blood cell activity. Simultaneously, it is hoped that these supplements can break down intravascular fat, accelerate vascular metabolism, and reduce the viscosity of intravascular lipids, thereby effectively preventing cardiovascular and cerebrovascular diseases.

[0003] Studies have found that approximately 20-30% of all men aged 18 to 55 experience sexual dysfunction, which significantly impacts their self-confidence and relationships with their partners. Currently, medication is the primary treatment for sexual dysfunction. However, some patients may experience a range of adverse reactions while using these medications, including fatigue, drowsiness, yawning, nausea, vomiting, dry mouth, diarrhea, and sweating. These unpleasant symptoms can often undermine a patient's confidence in treatment and may even lead them to discontinue treatment.

[0004] Salidroside is a natural active ingredient extracted from Rhodiola rosea, and it has attracted much attention due to its various biological activities. Studies have shown that salidroside has significant antioxidant, anti-inflammatory, and anti-apoptotic effects, and it has been widely used in traditional medicine to enhance physical strength, combat fatigue, and improve immunity.

[0005] Ergothioneine (EGT) is a natural amino acid derivative widely found in certain fungi, plants, and animals. In recent years, EGT has received increasing attention due to its potential antioxidant and cell-protective properties. Studies have shown that EGT can effectively scavenge free radicals, reduce oxidative stress-induced cell damage, and protect vascular endothelial cells from oxidative damage, thereby maintaining normal vascular function.

[0006] Therefore, combining the potential benefits of rhodioloside and ergothioneine to develop a supplement that can effectively enhance exercise protection or maintain cardiovascular health is of significant practical importance. This supplement would not only provide comprehensive cardiovascular protection but also significantly reduce the risk of cardiovascular disease through synergistic effects of multiple mechanisms. Currently, there are no reports on the combined use of rhodioloside and ergothioneine for protecting or maintaining cardiovascular health. Through further research and development, this supplement holds promise as a safe and effective cardiovascular health management tool, providing a new solution for cardiovascular disease prevention globally. Developing a supplement that can effectively improve sexual satisfaction or sexual health, or alleviate sexual dysfunction, is also of significant practical importance. Currently, there are no reports on the combined use of rhodioloside and ergothioneine for improving sexual satisfaction or sexual health. Through further research and development, this supplement holds promise as a means to promote reproductive health, improve the quality of sexual life, and maintain and enhance overall individual health and well-being. Summary of the Invention

[0007] To achieve the above objectives, in one aspect, the present invention provides a composition comprising rhodioloside or a pharmaceutically acceptable salt, ester, or derivative thereof; and ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof, the composition being used to protect or maintain cardiovascular health, to improve sexual satisfaction or sexual health or sexual ability, or to improve sexual dysfunction.

[0008] In some implementations, protecting or maintaining cardiovascular health includes preventing atherosclerosis, regulating the function of vascular endothelial cells, regulating vasomotor activity and permeability, improving myocardial function, improving vascular elasticity, and reducing hardening and calcification of the vascular wall; improving sexual satisfaction, sexual health, or sexual ability, or improving sexual dysfunction, including improving ejaculatory control, improving premature ejaculation, prolonging intravaginal ejaculation latency, improving erectile dysfunction, and improving decreased libido.

[0009] In some implementations, protecting or maintaining cardiovascular health is achieved by inhibiting the expression of vascular cell adhesion molecule-1 (VCAM-1) or intercellular adhesion molecule-1 (ICAM-1), or by increasing nitric oxide levels; improving sexual satisfaction or sexual health or sexual ability, or improving sexual dysfunction, by upregulating the expression of eNOS protein in the penile corpora cavernosa, or by increasing blood flow to the reproductive system and improving blood circulation.

[0010] In some implementations, compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed a reduction in VCAM-1 secretion levels of at least 10%, 20%, or 40%. Compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed a reduction in ICAM-1 secretion levels of at least 10%, 20%, or 30%. Compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed an increase in NO secretion levels of at least 10%, 20%, or 22%.

[0011] In some implementations, erectile rigidity (maximum ICP / MAP) and overall erectile quality (AUC) are restored by upregulating eNOS protein expression in the endothelial cells of the penis, thus improving sexual health. Compared with the untreated group, the eNOS protein expression in the endothelial cells of the penis was upregulated by at least 2-fold in the treated group. Compared with the untreated group, the maximum ICP / MAP ratio was increased by at least 1-fold, 1.5-fold, or 1.9-fold in the treated group. Compared with the untreated group, the area under the curve (AUC) of the ICP / MAP was increased by at least 1-fold, 1.5-fold, or 1.9-fold in the treated group.

[0012] In some embodiments, rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives are administered in doses of 1-2000 mg daily. In some embodiments, rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives may be administered in doses of 2-2000 mg, 5-1800 mg, 10-1500 mg, 15-1200 mg, 20-1000 mg, 25-800 mg, 30-500 mg, or 1-1000 mg, 5-800 mg, 5-300 mg, 5-100 mg, 10-500 mg, 10-300 mg, 10-100 mg, 15-100 mg, 20-80 mg daily.

[0013] In some embodiments, ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof is administered in doses of 1-1000 mg daily. In some embodiments, ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof may be administered in doses of 2-1000 mg, 5-800 mg, 8-500 mg, 10-300 mg, 15-200 mg, 20-100 mg, or 25-80 mg daily.

[0014] In some embodiments, the ratio of rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives to ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs, or derivatives is 1:50 to 500:1. In some embodiments, the ratio of rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives to ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs, or derivatives may be 1:20 to 300:1, 1:10 to 200:1, 1:5 to 100:1, 1:10 to 80:1, 1:1 to 80:1, 3:10 to 80:1, 3:1 to 50:1, 5:10 to 25:1, 5:1 to 25:1, 1:10 to 10:1, or 1:4 to 6:1.

[0015] In some embodiments, the composition is formulated as a nutritional supplement, food, beverage, or animal feed.

[0016] In some embodiments, the composition is in at least one of the following forms: suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, syrups, injections, and functionalized food compositions.

[0017] In some embodiments, the composition is prepared as a solid or liquid formulation.

[0018] In another aspect, the present invention provides a nutritional supplement, food, beverage, or animal feed comprising the composition described above.

[0019] In some embodiments, the composition is present in the nutritional supplement, food, beverage, or animal feed at a weight ratio of 0.1% to 95%. In some embodiments, the weight ratio of the composition in the nutritional supplement, food, beverage, or animal feed may be 0.5% to 90%, 1% to 85%, 5% to 80%, 10% to 75%, 15% to 70%, 20% to 65%, 25% to 60%, 30% to 55%, or 35% to 50%.

[0020] In some implementations, nutritional supplements, foods, beverages, and animal feeds also contain dietaryly or pharmaceutically acceptable carriers.

[0021] In another aspect, the present invention provides a method for protecting or maintaining cardiovascular health, for improving sexual satisfaction or sexual health or sexual ability, or for improving sexual dysfunction, the method comprising: administering a composition to a subject in need, the composition comprising rhodioloside or a pharmaceutically acceptable salt, ester or derivative thereof; and ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog or derivative thereof.

[0022] In some implementations, protecting or maintaining cardiovascular health includes preventing atherosclerosis, regulating the function of vascular endothelial cells, regulating vasomotor activity and permeability, improving myocardial function, improving vascular elasticity, and reducing hardening and calcification of the vascular wall; improving sexual satisfaction, sexual health, or sexual ability, or improving sexual dysfunction, including improving ejaculatory control, improving premature ejaculation, prolonging intravaginal ejaculation latency, improving erectile dysfunction, and improving decreased libido.

[0023] In some implementations, protecting or maintaining cardiovascular health is achieved by inhibiting the expression of vascular cell adhesion molecule-1 (VCAM-1) or intercellular adhesion molecule-1 (ICAM-1), or by increasing nitric oxide levels; improving sexual satisfaction or sexual health or sexual ability, or improving sexual dysfunction, by upregulating the expression of eNOS protein in the penile corpora cavernosa, or by increasing blood flow to the reproductive system and improving blood circulation.

[0024] In some implementations, compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed a reduction in VCAM-1 secretion levels of at least 10%, 20%, or 40%. Compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed a reduction in ICAM-1 secretion levels of at least 10%, 20%, or 30%. Compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed an increase in NO secretion levels of at least 10%, 20%, or 22%.

[0025] In some implementations, erectile rigidity (maximum ICP / MAP) and overall erectile quality (AUC) are restored by upregulating eNOS protein expression in the endothelial cells of the penis, thus improving sexual health. Compared with the untreated group, the eNOS protein expression in the endothelial cells of the penis was upregulated by at least 2-fold in the treated group. Compared with the untreated group, the maximum ICP / MAP ratio was increased by at least 1-fold, 1.5-fold, or 1.9-fold in the treated group. Compared with the untreated group, the area under the curve (AUC) of the ICP / MAP was increased by at least 1-fold, 1.5-fold, or 1.9-fold in the treated group.

[0026] In some embodiments, rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives are administered in doses of 1-2000 mg daily. In some embodiments, rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives may be administered in doses of 2-2000 mg, 5-1800 mg, 10-1500 mg, 15-1200 mg, 20-1000 mg, 25-800 mg, 30-500 mg, or 1-1000 mg, 5-800 mg, 5-300 mg, 5-100 mg, 10-500 mg, 10-300 mg, 10-100 mg, 15-100 mg, 20-80 mg daily.

[0027] In some embodiments, ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof is administered in doses of 1-1000 mg daily. In some embodiments, ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof may be administered in doses of 2-1000 mg, 5-800 mg, 8-500 mg, 10-300 mg, 15-200 mg, 20-100 mg, or 25-80 mg daily.

[0028] In some embodiments, the ratio of rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives to ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs, or derivatives is 1:50 to 500:1. In some embodiments, the ratio of rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives to ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs, or derivatives may be 1:20 to 300:1, 1:10 to 200:1, 1:5 to 100:1, 1:10 to 80:1, 1:1 to 80:1, 3:10 to 80:1, 3:1 to 50:1, 5:10 to 25:1, 5:1 to 25:1, 1:10 to 10:1, or 1:4 to 6:1.

[0029] In some implementations, the subjects are humans or mammals.

[0030] In some embodiments, the composition is administered orally, intravenously, intramuscularly, intraperitoneally, or sublingually.

[0031] In some implementations, application may be performed once daily or every other day, or at a suitable frequency. In some implementations, application may continue for a period of time, such as 3 days to 1 week, 2 weeks to 7 weeks, or 4 weeks to 14 weeks.

[0032] In some embodiments, the composition is formulated as a nutritional supplement, food, beverage, or animal feed.

[0033] In some embodiments, the composition is in at least one of the following forms: suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, syrups, injections, and functionalized food compositions.

[0034] In another aspect, the present invention provides the use of a composition in the preparation of a nutritional supplement, food, beverage, or animal feed for the protection or maintenance of cardiovascular health, for improving sexual satisfaction or sexual health or sexual ability, or for improving sexual dysfunction, the composition comprising rhodioloside or a pharmaceutically acceptable salt, ester, or derivative thereof; and ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof.

[0035] In some implementations, protecting or maintaining cardiovascular health includes preventing atherosclerosis, regulating the function of vascular endothelial cells, regulating vasomotor activity and permeability, improving myocardial function, improving vascular elasticity, and reducing hardening and calcification of the vascular wall; improving sexual satisfaction, sexual health, or sexual ability, or improving sexual dysfunction, including improving ejaculatory control, improving premature ejaculation, prolonging intravaginal ejaculation latency, improving erectile dysfunction, and improving decreased libido.

[0036] In some implementations, protecting or maintaining cardiovascular health is achieved by inhibiting the expression of vascular cell adhesion molecule-1 (VCAM-1) or intercellular adhesion molecule-1 (ICAM-1), or by increasing nitric oxide levels; improving sexual satisfaction or sexual health or sexual ability, or improving sexual dysfunction, by upregulating the expression of eNOS protein in the penile corpora cavernosa, or by increasing blood flow to the reproductive system and improving blood circulation.

[0037] In some implementations, compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed a reduction in VCAM-1 secretion levels of at least 10%, 20%, or 40%. Compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed a reduction in ICAM-1 secretion levels of at least 10%, 20%, or 30%. Compared with the ox-LDL-damaged group, the EGT and rhodioloside combination group showed an increase in NO secretion levels of at least 10%, 20%, or 22%.

[0038] In some implementations, erectile rigidity (maximum ICP / MAP) and overall erectile quality (AUC) are restored by upregulating eNOS protein expression in the endothelial cells of the penis, thus improving sexual health. Compared with the untreated group, the eNOS protein expression in the endothelial cells of the penis was upregulated by at least 2-fold in the treated group. Compared with the untreated group, the maximum ICP / MAP ratio was increased by at least 1-fold, 1.5-fold, or 1.9-fold in the treated group. Compared with the untreated group, the area under the curve (AUC) of the ICP / MAP was increased by at least 1-fold, 1.5-fold, or 1.9-fold in the treated group.

[0039] In some embodiments, rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives are administered in doses of 1-2000 mg daily. In some embodiments, rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives may be administered in doses of 2-2000 mg, 5-1800 mg, 10-1500 mg, 15-1200 mg, 20-1000 mg, 25-800 mg, 30-500 mg, or 1-1000 mg, 5-800 mg, 5-300 mg, 5-100 mg, 10-500 mg, 10-300 mg, 10-100 mg, 15-100 mg, 20-80 mg daily.

[0040] In some embodiments, ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof is administered in doses of 1-1000 mg daily. In some embodiments, ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof may be administered in doses of 2-1000 mg, 5-800 mg, 8-500 mg, 10-300 mg, 15-200 mg, 20-100 mg, or 25-80 mg daily.

[0041] In some embodiments, the ratio of rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives to ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs, or derivatives is 1:50 to 500:1. In some embodiments, the ratio of rhodioloside or its pharmaceutically acceptable salts, esters, or derivatives to ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs, or derivatives may be 1:20 to 300:1, 1:10 to 200:1, 1:5 to 100:1, 1:10 to 80:1, 1:1 to 80:1, 3:10 to 80:1, 3:1 to 50:1, 5:10 to 25:1, 5:1 to 25:1, 1:10 to 10:1, or 1:4 to 6:1.

[0042] In some embodiments, the composition is in at least one of the following forms: suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, syrups, injections, or functionalized food compositions. Attached Figure Description

[0043] Figure 1 shows the results of VCAM-1 secretion in each experimental group.

[0044] Figure 2 shows the results of ICAM-1 secretion in each experimental group.

[0045] Figure 3 shows the results of NO secretion in each experimental group.

[0046] Figure 4 shows the ejaculation latency of each experimental group.

[0047] Figure 5 shows the maximum ICP / MAP ratio for each experimental group.

[0048] Figure 6 shows the area under the ICP / MAP curves for each experimental group.

[0049] Figure 7 shows the relative protein expression levels of eNOS in the penis of each experimental group. Detailed Implementation

[0050] The present invention will now be further described with reference to preferred embodiments thereof. While the invention will be described in conjunction with preferred embodiments, it should be understood that they are not intended to limit the invention to these embodiments. Rather, the invention is intended to cover alternatives, modifications, and equivalents that may be included within the spirit and scope of the invention as defined in the claims.

[0051] As used herein, the term “or” is intended to include both “and” and “or”. In other words, the term “or” can also be replaced with “and / or”.

[0052] As used herein, unless the context clearly indicates otherwise, the singular forms “a / an” and “the” are intended to include the plural forms as well.

[0053] As used herein, the terms “comprising” or “including” or variations thereof refer to items that are included in the context of the term in their non-restrictive sense, but do not exclude items not specifically mentioned. It also includes the more restrictive verbs 'consistently composed of' and 'comprises of'.

[0054] As used herein, the terms “subject” or “mammal” are used interchangeably to refer to any animal to which the methods and compositions of this disclosure may be applied or administered. Animals may suffer from illness or other diseases, but they do not need to be sick to benefit from the methods and compositions of this disclosure. Therefore, any animal may utilize the disclosed compositions or become a recipient of the disclosed methods. Although the animal subject is preferably human, the methods and compositions of this invention are equally applicable to veterinary medicine, for example, for the treatment of domesticated species such as canines, felines, rodents, and various other pets; livestock such as cattle, horses, sheep, goats, pigs, etc.; and wild animals such as non-human primates in the wild or in zoos.

[0055] As used herein, the term "administration" refers to the process of delivering the disclosed composition or active ingredient to a subject. The compositions of the present invention are preferably administered via oral, intravenous, intramuscular, intraperitoneal, subcutaneous, topical, or sublingual routes, but may also be administered via other conventional routes to achieve the desired effect.

[0056] As used herein, the term "pharmaceutically acceptable" means pharmaceutically, physiologically, dietaryally, or cosmetically acceptable, and refers to combinations of compositions or reagents, materials or compositions and / or dosage forms thereof that are suitable for contact with human and animal tissues, compatible with other components of the composition, without excessive toxicity, irritation, allergic reactions or other problems or complications, and commensurate with a reasonable benefit / risk ratio, within the bounds of reasonable medical judgment.

[0057] In some embodiments, the compositions of the present invention can be prepared together with dietaryly or pharmaceutically acceptable carriers. These carriers include non-toxic, compatible substances commonly used in health foods and dietary supplements, animal feeds, and pharmaceutical preparations, such as sugars, starches, cellulose and their derivatives, powdered tragacanth gum, malt, gelatin, talc, oils, glycols, polyols, esters, agar, alginic acid, pyrogen-free water, isotonic saline, petrolatum, silicone oil, olive oil, and palm wax.

[0058] In some embodiments, the compositions of the present invention may be administered together with other supplements, such as vitamins, minerals, nootropics, and other supplements known in the art.

[0059] Various forms and formulations of the compositions are considered. The compositions will be formulated as nutritional supplements or dietary supplements, (medical) foods, beverages, animal feeds, in liquid or solid form. For oral administration, the compositions of the present invention may be in any suitable form, including solutions, tablets, gel capsules, capsules, or alternative nutritional foods or supplements.

[0060] The dosage of the active ingredient in this invention depends on the specific formulation and form. The dosage can also vary depending on factors such as the subject's sensitivity, age, sex, weight, and specific response. One or more doses can be administered once or multiple times daily or at a suitable frequency for any period of time. For example, an effective dose can be administered daily for one day, several days, multiple days, or indefinitely. In some embodiments, administration can continue for several days or longer, such as weeks or months.

[0061] The following examples illustrate selected embodiments of the present invention and are not intended to limit the scope of the invention. Example 1

[0062] Take 1 part (by weight) of rhodioloside and 1 part (by weight) of ergothionein and mix them evenly in a three-dimensional mixer to obtain the composition described in Example 1. Example 2-18

[0063] Examples 2-18 were prepared in the same manner as in Example 1. The weight parts of rhodioloside and ergothioneine are shown in the table below:

[0064]

[0065] The nutritional supplements, foods, beverages, and animal feeds of the present invention comprise the compositions of the present invention and suitable dietaryally or pharmaceutically acceptable carriers. Example 19

[0066] Cell culture: HUVECs (human umbilical vein endothelial cells) were cultured in 75 cm² culture flasks under the following conditions: 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, 1% cell supplement and 88% endothelial cell basal medium, in a cell culture incubator at 37°C and 5% CO2.

[0067] Treatment of HUVECs: HUVECs were seeded in 12-well plates (50,000 cells / mL) and allowed to grow to approximately 80% confluence. They were then divided into the following groups and treated with the corresponding solutions for 1 hour: Group 1 (Control Group): Culture medium. Group 2 (ox-LDL damage group): Culture medium. Group 3 (Example 12): Culture medium containing EGT and rhodioloside in a 4:1 ratio, specifically EGT concentration of 2 μg / mL and rhodioloside concentration of 0.5 μg / mL. Group 4 (Example 1): Culture medium containing EGT and rhodioloside in a 1:1 ratio, specifically EGT concentration of 1.25 μg / mL and rhodioloside concentration of 1.25 μg / mL. Group 5 (Example 4): Culture medium containing EGT and rhodioloside in a 1:4 ratio, specifically EGT concentration of 0.5 μg / mL and rhodioloside concentration of 2 μg / mL. Group 1 cells were placed in a cell culture incubator for 24 hours without any damage treatment. Groups 2-5 were co-incubated with 100 µg / mL oxidized low-density lipoprotein (ox-LDL) for 24 hours. After 24 hours, the cell supernatant was collected.

[0068] VCAM-1 and ICAM-1 expression: The content of VCAM-1 and ICAM-1 in the cell culture supernatant of each group was detected using an ELISA kit according to the instructions provided by the supplier.

[0069] NO determination: NO production is indirectly determined by the Griess reaction, that is, by measuring the concentration of stable end product nitrite in the supernatant of treated endothelial cells.

[0070] Statistical analysis: Data analysis was performed using Graphpad Prism 10 software and the two-way ANOVA statistical method.

[0071] Figure 1 shows the results of VCAM-1 secretion in each group. As shown in Figure 1, compared with group 2 (ox-LDL damage), groups 3-5 (combination of EGT and rhodioloside) significantly reduced VCAM-1 secretion levels, decreasing by 42.1%, 44.7%, and 50%, respectively. Statistical analysis of groups 2 and 3-5 showed p < 0.001 for all groups. Furthermore, this invention also explored the effects of EGT alone (2.5 μg / mL), rhodioloside alone (2.5 μg / mL), and rhodioloside + vitamin E (2 μg / mL + 0.5 μg / mL, with vitamin E being a commonly used positive control in this model). Compared with group 2 (ox-LDL damage), the VCAM-1 level decreased by approximately 35%, but the effect was not as good as the combination of EGT and rhodioloside. Figure 2 shows the results of ICAM-1 secretion in each group. As shown in Figure 2, compared with group 2 (ox-LDL damage), groups 3-5 (combination of EGT and rhodioloside) significantly reduced ICAM-1 secretion levels, decreasing by 34.7%, 32.6%, and 36.7%, respectively. Statistical analysis of groups 2 and 3-5 showed p < 0.0001 for all groups. Furthermore, this invention also explored the effects of EGT alone, rhodioloside alone, and rhodioloside + vitamin E groups. Compared with group 2 (ox-LDL damage), ICAM-1 levels decreased by approximately 26%, all of which were less than the combination of EGT and rhodioloside. Figure 3 shows the NO secretion results for each group. As shown in Figure 3, compared with group 2 (ox-LDL damage), groups 3-5 (combination of EGT and rhodioloside) significantly increased NO secretion levels, increasing by 23.7%, 22.7%, and 25.7%, respectively. Statistical analysis of groups 2 and 3-5 showed p < 0.01 for all groups. In addition, this invention also explored the effects of EGT alone, rhodioloside alone, and rhodioloside + vitamin E group. Compared with group 2 (ox-LDL damage), NO secretion increased by about 17%, and the effects were not as good as the combination of EGT + rhodioloside.

[0072] The results showed that the composition provided by this invention can work synergistically through multiple mechanisms. The combination of EGT and rhodioloside can resist ox-LDL-induced inflammatory responses (inhibiting the secretion of ICAM-1 and VCAM-1), thereby reducing leukocyte infiltration into the blood vessel wall and delaying or preventing the formation of early lipid streaks and plaques in atherosclerosis. Simultaneously, it protects or restores the ability of endothelial cells to synthesize NO, alleviates endothelial dysfunction, and is beneficial for restoring normal vasodilatory function, anti-inflammatory state, and anti-thrombotic capacity, thus providing a certain protective effect on the cardiovascular system. Example 20

[0073] Animals and Model Establishment: Forty 6-week-old male SD rats with normal erectile function, weighing 200 ± 20 g, were randomly divided into a control group, a model group, and experimental groups 1, 2, and 3, with 8 rats in each group. The control group was fed a standard diet with a fat content not exceeding 5% and a fat calorie content not exceeding 15%. The model and experimental groups were fed a high-fat diet with a fat content not exceeding 35% and a fat calorie content not exceeding 60%. After 8 weeks of feeding, all rats were fasted overnight. The model and experimental groups were intraperitoneally injected with 50 mg / kg streptozotocin (dissolved in 1% 0.1 mol / L citrate-sodium citrate buffer solution) to damage pancreatic β cells; the control group was intraperitoneally injected with an equal volume of citrate-sodium citrate buffer solution. One week after injection, an oral glucose tolerance test (OGTT) was performed to screen rats with a fasting blood glucose (FBG) ≥ 16.7 mmol / L. The selected rats were then placed in an observation box to acclimatize to the environment for 10 minutes. The room was kept quiet with the lights dimmed to just the right level for observation. Then, apomorphine 80 pg / kg was subcutaneously injected into the dorsal side of the neck of the rats. The apomorphine was dissolved in 0.5 mg / kg of vitamin C and physiological saline, and the volume was adjusted to 5 mL / kg. Each rat was observed for 30 minutes immediately after injection. If no erection was achieved, it was considered a diabetic rat model of erectile dysfunction (ED). Rats with erectile dysfunction were selected for subsequent drug intervention.

[0074] Animal intervention: Table 1 shows the intervention methods for each group of animals. The rats were administered the above doses by gavage for 12 weeks, followed by sexual behavior tests, erectile dysfunction assessment, and collection of penile tissue to detect eNOS (endothelial nitric oxide synthase) levels. Table 1. Intervention methods for each group of animals

[0075]

[0076] Screening for erectile dysfunction (ED) in diabetic rats: Rats underwent an erection test at a fixed time at night in a dimly lit, quiet, and undisturbed environment. After recording the rats' weight, they were placed in test cages and allowed to acclimatize for 10 minutes. Apomorphine 100 μg / kg was then injected subcutaneously into the neck. The rats were observed for 30 minutes, and penile erection was recorded. Penile erection was defined as penile engorgement with engorgement of the glans penis and the appearance of the penile shaft, counted as one erection. A positive erection test was considered to have ≥1 erection; otherwise, it was considered negative. No response (penile erection or exposure of the glans penis) indicated successful induction of diabetic erectile dysfunction. Rats with a response indicated induction failure and were not used for subsequent drug intervention.

[0077] Sexual behavior test: In a low-light and quiet environment, one male mouse was placed in a cage with one female mouse that was artificially induced to estrus (30 μg estradiol benzoate was injected subcutaneously into the neck of each mouse 48 hours before the experiment; 500 μg progesterone was injected subcutaneously into the neck of each mouse 4-6 hours before the experiment). After being placed in the cage, the mating behavior of the male mouse was observed and recorded within 30 minutes. The main observation was the ejaculation latency period (the time interval from the first insertion to the occurrence of ejaculation).

[0078] Erectile Dysfunction Assessment: Penile erection depends on the increase in intracavernosal pressure (ICP) caused by engorgement of the cavernous sinuses. Therefore, measuring ICP during penile erection can accurately quantify erectile function, while monitoring arterial pressure (AP) can eliminate the interference of systemic blood pressure fluctuations on ICP. After anesthetizing rats with 0.4% sodium pentobarbital (20 mg / kg), the BL-420F biosignal acquisition system (Techman Soft, Chengdu, China) was activated to apply electrical stimulation (15 Hz, 5 V, 1 min) to the cavernous nerves to measure and record ICP and mean arterial pressure (MAP). Erectile function was assessed by calculating the maximum ICP / MAP ratio and the area under the ICP / MAP ratio (AUC). After measurement, penile tissue from dissected rats was used for further protein analysis.

[0079] eNOS protein level detection: Total protein was extracted from the penis, and the eNOS protein level in the penis was detected by Western blot. An internal control protein (β-actin) was used to correct for loading errors. Protein bands were visualized using horseradish peroxidase (HRP)-labeled goat anti-rabbit / mouse antibody, with signal enhancement using Western ECL chemiluminescence solution, and imaging was performed using a ChemiDoc MP imaging system. The gray values ​​of the target bands were analyzed using ImageJ software, and the relative protein expression level was expressed as the gray ratio of eNOS to β-actin.

[0080] Statistical analysis: Data analysis was performed using Graphpad Prism 10 software. The results showed that the application of the composition provided by the present invention significantly upregulated the expression of eNOS protein in the endothelial cells of the penile cavernous body, thereby restoring erectile rigidity (maximum ICP / MAP) and overall erectile quality (AUC) to a certain extent and improving the sexual health of rats.

[0081] Figure 4 shows the ejaculation latency of each group. Diabetic ED mice have erectile and ejaculatory dysfunction, resulting in a longer latency period. The composition of this invention can effectively shorten the latency period. As shown in Figure 4, compared with the model group, experimental groups 1-3 significantly shortened the ejaculation latency period, by 16.7%, 18.8%, and 22.3%, respectively. Statistical analysis showed that the p-values ​​for both the model group and the experimental group were <0.05. In addition, this invention also explored the effects of EGT alone, rhodioloside alone, and rhodioloside + sildenafil. Compared with the model group, the ejaculation latency period was shortened by about 14 times, but none of these were as good as the combination of EGT + rhodioloside. Figure 5 shows the maximum ICP / MAP ratio of each group. As shown in Figure 5, compared with the model group, experimental groups 1-3 significantly increased the maximum ICP / MAP ratio, by 1.9 times, 2 times, and 2 times, respectively. Statistical analysis showed that the p-values ​​for both the model group and the experimental group were <0.05. Furthermore, this invention also investigated the effects of EGT alone (30 mg / kg), rhodioloside alone (30 mg / kg), and rhodioloside + sildenafil (24 mg / kg + 6 mg / kg). Compared with the model group, the maximum ICP / MAP ratio increased by approximately 1.6 times, but none of these were as significant as the combination of EGT and rhodioloside. Figure 6 shows the area under the ICP / MAP curve for each group. As shown in Figure 6, compared with the model group, experimental groups 1-3 significantly increased the area under the ICP / MAP curve (AUC), increasing by 1.95 times, 1.9 times, and 2 times, respectively. Statistical analysis showed that the p-values ​​for both the model group and the experimental group were <0.01. In addition, this invention also investigated the effects of EGT alone, rhodioloside alone, and rhodioloside + sildenafil. Compared with the model group, the area under the ICP / MAP curve increased by approximately 1.7 times, but none of these were as significant as the combination of EGT and rhodioloside. Figure 7 shows the relative eNOS protein expression levels in the penis of each group. As shown in Figure 7, compared with the model group, experimental groups 1-3 significantly upregulated the relative protein expression level of eNOS in the penis, by 2.26-fold, 2.22-fold, and 2.3-fold, respectively. Statistical analysis showed that the p-values ​​for both the model group and the experimental group were <0.05. Furthermore, this invention also explored the effects of EGT alone, rhodioloside alone, and rhodioloside + sildenafil. Compared with the model group, the relative protein expression level of eNOS in the penis was upregulated by approximately 2-fold, but none of these effects were as significant as the combination of EGT and rhodioloside.

[0082] While specific embodiments and examples of the invention have been described herein, those skilled in the art will understand that any modifications and variations can be made without departing from the principles of the invention. The above embodiments and descriptions do not limit the scope of the invention. Any combination of embodiments of the invention, as well as any obvious extensions or analogies thereof, are within the scope of the invention. Furthermore, the invention covers any arrangement intended to achieve the same purpose, and all such variations and modifications falling within the scope of the appended claims.

Claims

1. A composition, characterized in that, The composition comprises rhodioloside or a pharmaceutically acceptable salt, ester, or derivative thereof; and ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof, and the composition is intended to protect or maintain cardiovascular health, to improve sexual satisfaction or sexual health or sexual ability, or to improve sexual dysfunction.

2. The composition according to claim 1, characterized in that, The protection or maintenance of cardiovascular health includes preventing atherosclerosis, regulating the function of vascular endothelial cells, regulating vasomotor activity and permeability, improving myocardial function, improving vascular elasticity, and reducing hardening and calcification of the vascular wall; the improvement of sexual satisfaction, sexual health, or sexual ability, or improvement of sexual dysfunction, includes improving ejaculatory control, improving premature ejaculation, prolonging intravaginal ejaculation latency, improving erectile dysfunction, improving erectile hardness, and improving decreased libido.

3. The composition according to claim 1 or 2, characterized in that, The protection or maintenance of cardiovascular health is achieved by inhibiting the expression of vascular cell adhesion molecule-1 (VCAM-1) or intercellular adhesion molecule-1 (ICAM-1), or by increasing the content of nitric oxide; sexual health or sexual ability is improved, or sexual dysfunction is improved, by upregulating the expression of eNOS protein in the endothelial cells of the penile cavernous body, or by increasing blood flow to the reproductive system and improving blood circulation.

4. The composition according to any one of claims 1 to 3, characterized in that, The rhodioloside or its pharmaceutically acceptable salts, esters or derivatives are administered at a dose of 1-2000 mg per day.

5. The composition according to any one of claims 1 to 4, characterized in that, The ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs or derivatives are administered in doses of 1-1000 mg per day.

6. The composition according to any one of claims 1 to 5, characterized in that, The ratio of the rhodioloside or its pharmaceutically acceptable salt, ester or derivative to the ergothioneine or its pharmaceutically acceptable salt, acid, ester, analog or derivative is 1:50 to 500:

1.

7. The composition according to any one of claims 1 to 6, characterized in that, The composition is formulated into nutritional supplements, food, beverages, and animal feed.

8. The composition according to any one of claims 1 to 7, characterized in that, The composition is in at least one of the following forms: suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, syrups, injections, and functional food compositions.

9. A method for protecting or maintaining cardiovascular health, for improving sexual satisfaction or sexual health or sexual ability, or for improving sexual dysfunction, characterized in that, The method includes administering a composition to a subject in need, the composition comprising rhodioloside or a pharmaceutically acceptable salt, ester, or derivative thereof; and ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof.

10. The method according to claim 9, characterized in that, The protection or maintenance of cardiovascular health includes preventing atherosclerosis, regulating the function of vascular endothelial cells, regulating vasomotor activity and permeability, improving myocardial function, improving vascular elasticity, and reducing hardening and calcification of the vascular wall; the improvement of sexual satisfaction, sexual health, or sexual ability, or improvement of sexual dysfunction, includes improving ejaculatory control, improving premature ejaculation, prolonging intravaginal ejaculation latency, improving erectile dysfunction, improving erectile hardness, and improving decreased libido.

11. The method according to claim 9 or 10, characterized in that, The protection or maintenance of cardiovascular health is achieved by inhibiting the expression of vascular cell adhesion molecule-1 (VCAM-1) or intercellular adhesion molecule-1 (ICAM-1), or by increasing the content of nitric oxide; sexual health or sexual ability is improved, or sexual dysfunction is improved, by upregulating the expression of eNOS protein in the endothelial cells of the penile cavernous body, or by increasing blood flow to the reproductive system and improving blood circulation.

12. The method according to any one of claims 9 to 11, characterized in that, The rhodioloside or its pharmaceutically acceptable salts, esters or derivatives are administered at a dose of 1-2000 mg per day.

13. The method according to any one of claims 9 to 12, characterized in that, The ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs or derivatives are administered in doses of 1-1000 mg per day.

14. The method according to any one of claims 9 to 13, characterized in that, The ratio of the rhodioloside or its pharmaceutically acceptable salt, ester or derivative to the ergothioneine or its pharmaceutically acceptable salt, acid, ester, analog or derivative is 1:50 to 500:

1.

15. The method according to any one of claims 9 to 14, characterized in that, The subjects are humans or mammals.

16. The method according to any one of claims 9 to 15, characterized in that, The composition is administered orally, intravenously, intramuscularly, intraperitoneally, or sublingually.

17. The method according to any one of claims 9 to 16, characterized in that, The composition is formulated into nutritional supplements, food, beverages, and animal feed.

18. The method according to any one of claims 9 to 17, characterized in that, The composition is in at least one of the following forms: suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, syrups, injections, and functional food compositions.

19. The use of a composition in the preparation of nutritional supplements, foods, beverages, or animal feeds for the protection or maintenance of cardiovascular health, for improving sexual satisfaction, sexual health, or sexual ability, or for improving sexual dysfunction, characterized in that, The composition comprises rhodioloside or a pharmaceutically acceptable salt, ester, or derivative thereof; and ergothioneine or a pharmaceutically acceptable salt, acid, ester, analog, or derivative thereof.

20. The use according to claim 19, characterized in that, The protection or maintenance of cardiovascular health includes preventing atherosclerosis, regulating the function of vascular endothelial cells, regulating vasomotor activity and permeability, improving myocardial function, improving vascular elasticity, and reducing hardening and calcification of the vascular wall; the improvement of sexual satisfaction, sexual health, or sexual ability, or improvement of sexual dysfunction, includes improving ejaculatory control, improving premature ejaculation, prolonging intravaginal ejaculation latency, improving erectile dysfunction, improving erectile hardness, and improving decreased libido.

21. The use according to claim 19 or 20, characterized in that, The protection or maintenance of cardiovascular health is achieved by inhibiting the expression of vascular cell adhesion molecule-1 (VCAM-1) or intercellular adhesion molecule-1 (ICAM-1), or by increasing the content of nitric oxide; sexual health or sexual ability is improved, or sexual dysfunction is improved, by upregulating the expression of eNOS protein in the endothelial cells of the penile cavernous body, or by increasing blood flow to the reproductive system and improving blood circulation.

22. The use according to any one of claims 19 to 21, characterized in that, The rhodioloside or its pharmaceutically acceptable salts, esters or derivatives are administered at a dose of 1-2000 mg per day.

23. The use according to any one of claims 19 to 22, characterized in that, The ergothioneine or its pharmaceutically acceptable salts, acids, esters, analogs or derivatives are administered in doses of 1-1000 mg per day.

24. The use according to any one of claims 19 to 23, characterized in that, The ratio of the rhodioloside or its pharmaceutically acceptable salt, ester or derivative to the ergothioneine or its pharmaceutically acceptable salt, acid, ester, analog or derivative is 1:50 to 500:

1.

25. The use according to any one of claims 19 to 24, characterized in that, The composition is in at least one of the following forms: suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, syrups, injections, and functional food compositions.

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