Capsule containing supramolecular anti-aging factor and used for improving ovarian function and preparation method thereof

Abstract

The invention provides a capsule containing supramolecular anti-aging factors and used for improving ovarian functions and a preparation method of the capsule. The capsule is prepared from the following components by weight: 0.4 to 0.8 g of ergothioneine, 5 to 8 g of fruit polyphenol, 10 to 20 g of inositol composition, 5 to 8 g of diindolylmethane, 0.01 to 0.02 g of folic acid and 100 to 200 g of auxiliary materials. High-purity ergothioneine is extracted from pleurotus eryngii through microwave and ultrasonic combination, chiral inositol is extracted from tartary buckwheat seeds, the chiral inositol and myo-inositol are maintained to be in a proper proportion, meanwhile, fruit polyphenol and a small amount of folic acid are used in a matched mode, gelatin and other auxiliary materials are used for preparing a capsule form, the capsule is more convenient to take, the symptoms of premature ovarian failure can be relieved, and the effect of preventing and treating premature ovarian failure is achieved. The raw materials for preparing the capsule are environment-friendly, and the preparation process is simple and suitable for large-scale production.

Description

Technical Field

[0001] The invention belongs to the field of capsules, and relates to a capsule containing a supramolecular anti-aging factor for improving ovarian function and a preparation method thereof. Background Art

[0002] As a key organ in the female reproductive system, the function of the ovaries gradually declines with age. During this process, the number of follicles in the ovaries decreases, the quality of oocytes declines, and other phenomena occur, which in turn lead to irregular menstruation, insufficient ovarian hormone secretion, and cessation of ovulation, which ultimately affect women's fertility and cause various diseases. Among them, polycystic ovary syndrome is a common endocrine disease related to reproductive and metabolic disorders in women. It is closely related to reproductive and metabolic disorders and affects about 6-20% of women worldwide.

[0003] PCOS not only causes typical symptoms such as reduced fertility, irregular menstruation, obesity and hirsutism in women of childbearing age, but also causes a series of metabolic problems. If PCOS is not treated promptly and effectively, the health risks faced by patients will increase significantly, including serious diseases such as type 2 diabetes, cardiovascular disease, metabolic syndrome and endometrial cancer.

[0004] At present, the drug treatment used in clinical practice is mainly based on the diagnostic criteria of PCOS patients, and anti-androgen drugs are used to treat PCOS patients of childbearing age for patients with endocrine abnormalities. Such anti-androgen drugs will have side effects on the body after long-term use and are potentially carcinogenic, so it is particularly important to develop natural, safe and side-effect-free products to improve ovarian function.

[0005] Ergothioneine is a natural chiral amino acid strong antioxidant, with free radical scavenging, detoxification, maintaining the biosynthesis of DNA, the normal growth of cells, cellular immunity and anti-aging and other functions, especially plays an important role in anti-oxidation and energy regulation, and is a multifunctional cell physiological protective agent. And the preparation method of ergothioneine is various, with the advantages of low cost, easy acquisition of raw materials, easy expansion of output, but the product for improving ovarian function based on ergothioneine is not many at present. Therefore, developing a product for improving ovarian function containing ergothioneine is a problem to be solved in the present invention. Summary of the invention

[0006] The object of the present invention is that, on the one hand, the present invention provides a capsule containing a supramolecular anti-aging factor for improving ovarian function, wherein the capsule comprises the following components by weight: 0.4-0.8g of ergothioneine, 5-8g of fruit polyphenols, 10-20g of inositol composition, 5-8g of diindolylmethane, 0.01-0.02g of folic acid and 100-200g of auxiliary materials.

[0007] In order to enhance the anti-aging function of the ovaries, the anti-aging factor ergothioneine is added to the capsule containing supramolecular anti-aging factors for improving ovarian function.

[0008] As a further improvement, the preparation method of the ergothioneine comprises the following steps:

[0009] S2-1: taking fresh Pleurotus eryngii and placing it in a refrigerator to rapidly cool and freeze it, and then performing vacuum freeze drying to a constant weight, and then using a grinder to fully grind it into powder to obtain Pleurotus eryngii powder;

[0010] S2-2: Take the Pleurotus eryngii powder obtained in step S2-1 and the ethanol solution in a solid-liquid ratio of 1: (20-60)

[0011] (g / mL) is mixed, then treated with microwave and ultrasound, and then centrifuged and dried to obtain ergothioneine.

[0012] As a further improvement, the volume fraction of the ethanol solution in step S2-2 is 50%-75%.

[0013] As a further improvement, in step S2-2, the power of the microwave is 100-700 W, the power of the ultrasound is 200-600 W, the microwave time is 5-20 min, and the ultrasound time is 10-20 min.

[0014] As a further improvement, the inositol composition comprises chiral inositol and myo-inositol, and the mass ratio of the chiral inositol to myo-inositol is 1:(35-45).

[0015] As a further improvement, the preparation method of the chiral inositol comprises the following steps:

[0016] S6-1: drying the tartary buckwheat seeds in an oven to constant weight, crushing them with an intermittent high-speed grinder, passing them through a 70-90 mesh sieve to obtain tartary buckwheat seed powder, and sealing them in a dryer for later use;

[0017] S6-2: Weigh buckwheat seed powder into a beaker, add distilled water at a solid-liquid ratio of 1:(10-30) (g / mL), adjust the pH to 5-6, add β-galactosidase, and perform enzymolysis at 30-50°C for 0.5-1h. After enzymolysis, add ethanol solution, and then place in a constant temperature water bath for ultrasonic oscillation extraction, centrifuge at 2000-3500r / min, take the supernatant, and freeze-dry and purify to obtain chiro-inositol.

[0018] As a further improvement, the fruit polyphenols include one or more of pomegranate polyphenols, cherry polyphenols, grape polyphenols, mangosteen polyphenols, blackcurrant polyphenols, blueberry polyphenols and apple polyphenols.

[0019] As a further improvement, the auxiliary material comprises one or more of gelatin, chitosan and gum arabic.

[0020] On the other hand, the present invention also provides a method for preparing a capsule containing supramolecular anti-aging factors for improving ovarian function, comprising the following steps:

[0021] S9-1: Weigh ergothioneine, fruit polyphenols, inositol composition, diindolylmethane and folic acid, prepare a sample solution with anhydrous ethanol, wrap it with tin foil and store it for later use; weigh auxiliary materials, prepare auxiliary material solution with purified water, and store it at low temperature for later use;

[0022] S9-2: At a temperature of 30-45°C, add the auxiliary material solution into a beaker, then drop the sample solution while stirring, then adjust the pH value, continue stirring for a few minutes, then cool to room temperature, add the curing agent under ice bath conditions, and continue stirring to obtain the intermediate; finally, centrifuge the intermediate, purify it, and freeze-dry it to obtain capsules containing supramolecular anti-aging factors for improving ovarian function.

[0023] As a further improvement, in step S9-2, the curing temperature is 0-6°C.

[0024] The beneficial effects of the present invention are:

[0025] The present invention provides a capsule containing supramolecular anti-aging factors for improving ovarian function. The capsule adopts microwave ultrasound combined extraction technology to extract high-purity ergothioneine from Pleurotus eryngii, effectively remove excess oxygen free radicals in the body, prevent oocyte apoptosis, repair ovarian damage, reduce follicle loss, promote follicle development, increase the number of eggs, and help to postpone amenorrhea. In addition, the capsule also contains chiro-inositol extracted from bitter buckwheat seeds, by maintaining the appropriate ratio of chiro-inositol to myo-inositol, regulating estrogen balance, assisting polycystic ovary syndrome patients to restore natural ovulation, improve irregular menstruation, and promote ovarian health. At the same time, combined with the formula of fruit polyphenols and folic acid, the capsule form is made of excipients such as gelatin, which is more convenient to take and helps to alleviate the symptoms of premature ovarian failure. The raw material source of the capsule is environmentally friendly, the preparation process is simple, and it is suitable for large-scale production. DETAILED DESCRIPTION

[0026] The present invention will be described below in conjunction with specific implementation schemes. It should be noted that the following examples are examples of the present invention and are only used to illustrate the present invention, not to limit the present invention. Without departing from the subject matter or scope of the present invention, other combinations and various improvements within the concept of the present invention may be made. In the following examples, the compound monomers and related reagents used can all be purchased from the market, among which Pleurotus eryngii is purchased from the farmer's market, and fresh and complete ones are selected; pomegranate polyphenols, grape polyphenols, cherry polyphenols, blackcurrant polyphenols, blueberry polyphenols, and mangosteen polyphenols are purchased from Xi'an Dongchi Biotechnology Co., Ltd.; commercial ergothioneine is purchased from Shaanxi Sanhe Pharmaceutical Co., Ltd. with a purity of 99%; gelatin is purchased from Henan Guangheng Food Ingredients Co., Ltd. with a CAS number of 9000-70-8; chitosan is purchased from Shanghai Gaoming Chemical Co., Ltd. with a CAS number of 9012-76-4; gum arabic is purchased from Henan Guangheng Food Ingredients Co., Ltd. with a CAS number of 9000-01-5.

[0027] The capsules containing supramolecular anti-aging factors for improving ovarian function prepared in the following Examples 1-4 and Comparative Examples 1-3 all include the following steps:

[0028] Preparation of Ergothioneine AC:

[0029] S2-1: 100 g of fresh Pleurotus eryngii was placed in a refrigerator for 12 h to rapidly cool and freeze, and then vacuum freeze-dried to constant weight (cold trap temperature -57°C, vacuum degree 200 Pa), and then fully crushed into powder using a pulverizer to obtain Pleurotus eryngii powder;

[0030] S2-1: The Pleurotus eryngii powder prepared in step S2-1 is mixed with the ethanol solution according to the contents in Table 1, and then subjected to microwave and ultrasonic treatment. The microwave and ultrasonic conditions are set as shown in Table 1.

[0031] The crude product was obtained, and then centrifuged at a speed of 8000r / min for 10min, the supernatant was taken, and freeze-dried to obtain ergothioneine.

[0032] Table 1

[0033]

[0034]

[0035] Preparation of chiro-inositol:

[0036] S6-1: drying the tartary buckwheat seeds in an oven at 60° C. to constant weight, crushing them with an intermittent high-speed grinder, passing them through an 80-mesh sieve to obtain tartary buckwheat seed powder, and sealing them in a desiccator for later use;

[0037] S6-2: Weigh 5 g of the bitter buckwheat seed powder obtained in step S6-1 into a beaker, add 50 mL of distilled water at a solid-liquid ratio of 1:10 (g / mL), adjust the pH value to 6, add 0.1 g of galactosidase, and perform enzymolysis at 30°C for 0.5 h. After enzymolysis, add a 50% ethanol solution by volume, and then place it in a constant temperature water bath at 50°C for ultrasonic oscillation extraction for 0.5 h, with an ultrasonic power of 150 W, and then centrifuge at a speed of 3000 r / min, take the supernatant, and freeze-dry to obtain chiro-inositol.

[0038] Preparation of capsules containing supramolecular anti-aging factors for improving ovarian function:

[0039] S9-1: Weigh thioneine, fruit polyphenols, inositol composition, diindolylmethane and folic acid, prepare a 1 mg / mL sample solution with anhydrous ethanol, wrap it with tin foil and store it for later use; weigh the auxiliary materials, prepare a 20 mg / mL auxiliary material solution with purified water, and store it at low temperature for later use;

[0040] S9-2: At a temperature of 30-45°C, add 10 mL of the auxiliary material solution into a beaker, and then add 5 mL of the sample solution dropwise while stirring. Then adjust the pH value to the isoelectric point of the auxiliary material solution, continue stirring for a few minutes, then cool to room temperature, add to the curing agent solution under ice bath conditions, keep the temperature at 0-6°C during curing, and continue stirring for 30 minutes to obtain an intermediate; finally, the intermediate is centrifuged and purified, and freeze-dried for 24 hours to obtain a capsule containing a supramolecular anti-aging factor for improving ovarian function. The curing agent solution is prepared by adding calcium chloride and ascorbic acid in a mass ratio of 6.5:1 to an acetic acid-sodium acetate buffer solution at a pH of 4, and stirring and dissolving evenly to obtain the capsule.

[0041] Example 1 provides a capsule containing a supramolecular anti-aging factor for improving ovarian function, comprising the following components: 0.6 g of ergothioneine A, 6 g of fruit polyphenols, 15 g of inositol composition, 6 g of diindolylmethane, 0.013 g of folic acid and 150 g of gelatin, wherein the fruit polyphenols comprise pomegranate polyphenols, cherry polyphenols and grape polyphenols in a mass ratio of 1:1:1, and the mass ratio of chiral inositol to myo-inositol in the inositol composition is 1:40.

[0042] Example 2 provides a capsule containing a supramolecular anti-aging factor for improving ovarian function, comprising the following components: 0.5 g of ergothioneine B, 5 g of fruit polyphenols, 16 g of inositol composition, 5 g of diindolylmethane, 0.015 g of folic acid and 100 g of chitosan, wherein the fruit polyphenols comprise pomegranate polyphenols, blackcurrant polyphenols, blueberry polyphenols and apple polyphenols in a mass ratio of 1:2:1:1, and the mass ratio of chiral inositol to myo-inositol in the inositol composition is 1:35.

[0043] Example 3 provides a capsule containing a supramolecular anti-aging factor for improving ovarian function, comprising the following components: 0.8 g of ergothioneine C, 8 g of fruit polyphenols, 20 g of inositol composition, 8 g of diindolylmethane, 0.018 g of folic acid and 180 g of gum arabic, wherein the fruit polyphenols comprise pomegranate polyphenols, blueberry polyphenols, mangosteen polyphenols and apple polyphenols in a mass ratio of 1:1:2:1, and the mass ratio of chiral inositol to myo-inositol in the inositol composition is 1:45.

[0044] Example 4 provides a capsule containing supramolecular anti-aging factors for improving ovarian function. The components and contents are basically the same as those in Example 1, except that the mass ratio of chiral inositol to myo-inositol in the inositol composition is 1:50.

[0045] Comparative Example 1 provides a capsule containing a supramolecular anti-aging factor for improving ovarian function, and the components and contents are substantially the same as those in Example 1, except that the 0.6 g of ergothioneine A is replaced with 1 g of ergothioneine A.

[0046] Comparative Example 2 A capsule containing a supramolecular anti-aging factor for improving ovarian function, the components and contents are substantially the same as those in Example 1, except that the 0.6 g of ergothioneine A is replaced with 0.2 g of ergothioneine A.

[0047] Comparative Example 3 A capsule containing a supramolecular anti-aging factor for improving ovarian function, the components and contents are substantially the same as those in Example 1, except that the 0.6 g of ergothioneine A is replaced with 0.6 g of commercial ergothioneine.

[0048] The test method is as follows:

[0049] Select several SD female rats weighing 180-220g. After 1 week of adaptive feeding, vaginal exfoliated cell smear examination was performed once a day for 10 consecutive days, and 90 rats with an estrus period of 4-5 days were screened out. Randomly divided into a blank control group, a model control group, Example 1, Example 2, Example 3, Example 4 and Comparative Example 1, Comparative Example 2, Comparative Example 3, 10 rats in each group. Modeling was performed on rats with normal estrous cycle with reference to the modeling method of the Second Affiliated Hospital of Sun Yat-sen University of Medical Sciences. Except for the blank control group given intraperitoneal injection of 1mL of normal saline, other groups were given intraperitoneal injection of cyclophosphamide 75mg / kg, once a day, for 4 consecutive weeks. At the same time, oral administration was performed according to the dose, 10mL / kg, once a day, for 4 consecutive weeks, and the blank control group and the model control group were given equal volumes of distilled water.

[0050] After the experiment, the ovaries were weighed to calculate the index; serum E2 and FSH were measured to evaluate the ovarian reserve function and observe the improvement of the ovarian function of the rats.

[0051] (1) Ovarian index: After the ovaries of rats were completely removed, the fascia and adipose tissue were removed and the ovaries were weighed using an electronic balance. The index was calculated according to the following formula: Ovarian index = ovarian wet weight (mg) / body weight (g) x 100%.

[0052] (2) Hormone detection: Blood was collected from the ear vein, and the serum was separated and stored at -20°C for testing. The changes in E2 and FSH levels in rat serum were determined by radioimmunoassay.

[0053] The test results are shown in Table 2.

[0054] Table 2

[0055]

[0056]

[0057] It can be seen from Examples 1-3 that the capsules provided by the present invention contain supramolecular anti-aging factors for improving ovarian function. High-purity ergothioneine is extracted from Pleurotus eryngii by microwave ultrasound, and chiral inositol extracted from buckwheat seeds is used in combination, fruit polyphenols and folic acid are added, and then the capsules are made into capsules by excipients such as gelatin. It can be seen from animal experiments that the capsules have the function of improving premature ovarian failure.

[0058] From the comparison between Example 1 and Comparative Examples 1-3, it can be seen that when the components provided by the present invention are within a reasonable range and the capsules prepared by the method provided by the present invention have a good improvement effect on premature ovarian failure in mice caused by cyclophosphamide in terms of ovarian index, E2 and FSH indicators.

[0059] In summary, the capsule containing supramolecular anti-aging factors for improving ovarian function provided by the present invention, through the preparation method provided by the present invention, high-purity ergothioneine is extracted from Pleurotus eryngii by combining microwave and ultrasound, chiral inositol is extracted from tartary buckwheat seeds, while ensuring that chiral inositol and myo-inositol are in a suitable ratio, fruit polyphenols and a small amount of folic acid are used in combination, and the capsules made of auxiliary materials such as gelatin have a certain therapeutic effect on premature ovarian failure.

[0060] The above implementation modes are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with this technology to understand the content of the present invention and implement it, and they cannot be used to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit of the present invention should be included in the protection scope of the present invention.

Claims

1. A capsule containing supramolecular anti-aging factors for improving ovarian function, characterized in that: The capsule comprises the following components by weight: 0.4-0.8 g of ergothioneine, 5-8 g of fruit polyphenols, 10-20 g of inositol composition, 5-8 g of diindolylmethane, 0.01-0.02 g of folic acid and 100-200 g of auxiliary materials.

2. The capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 1, characterized in that: The preparation method of described ergothioneine comprises the following steps: S2-1: taking fresh Pleurotus eryngii and placing it in a refrigerator to rapidly cool and freeze it, and then performing vacuum freeze drying to a constant weight, and then using a grinder to fully grind it into powder to obtain Pleurotus eryngii powder; S2-2: The Pleurotus eryngii powder obtained in step S2-1 is mixed with the ethanol solution at a solid-liquid ratio of 1:(20-60) (g / mL), and then subjected to microwave and ultrasonic treatment, and then centrifugal drying to obtain ergothioneine.

3. The capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 2, characterized in that: The volume fraction of the ethanol solution in step S2-2 is 50%-75%.

4. The capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 2, characterized in that: In step S2-2, the power of the microwave is 100-700 W, the power of the ultrasound is 200-600 W, the microwave time is 5-20 min, and the ultrasound time is 10-20 min.

5. The capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 1, characterized in that: The inositol composition comprises chiro-inositol and myo-inositol, and the mass ratio of the chiro-inositol to myo-inositol is 1:(35-45).

6. The capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 5, characterized in that: The preparation method of the chiral inositol comprises the following steps: S6-1: drying the tartary buckwheat seeds in an oven to constant weight, crushing them with an intermittent high-speed grinder, passing them through a 70-90 mesh sieve to obtain tartary buckwheat seed powder, and sealing them in a dryer for later use; S6-2: Weigh buckwheat seed powder into a beaker, add distilled water at a solid-liquid ratio of 1:(10-30) (g / mL), adjust the pH to 5-6, add β-galactosidase, and perform enzymolysis at 30-50°C for 0.5-1h. After enzymolysis, add ethanol solution, and then place in a constant temperature water bath for ultrasonic oscillation extraction, centrifuge at 2000-3500r / min, take the supernatant, and freeze-dry and purify to obtain chiro-inositol.

7. The capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 1, characterized in that: The fruit polyphenols include one or more of pomegranate polyphenols, cherry polyphenols, grape polyphenols, mangosteen polyphenols, blackcurrant polyphenols, blueberry polyphenols and apple polyphenols.

8. The capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 1, characterized in that: The auxiliary materials include one or more of gelatin, chitosan and gum arabic.

9. The method for preparing the capsule containing supramolecular anti-aging factors for improving ovarian function according to any one of claims 1 to 8, comprising the following steps: S9-1: Weigh ergothioneine, fruit polyphenols, inositol composition, diindolylmethane and folic acid, prepare a sample solution with anhydrous ethanol, wrap it with tin foil and store it for later use; weigh auxiliary materials, prepare auxiliary material solution with purified water, and store it at low temperature for later use; S9-2: At a temperature of 30-45°C, add the auxiliary material solution into a beaker, then drop the sample solution while stirring, then adjust the pH value, continue stirring for a few minutes, then cool to room temperature, add the curing agent under ice bath conditions, and continue stirring to obtain the intermediate; finally, centrifuge the intermediate, purify it, and freeze-dry it to obtain capsules containing supramolecular anti-aging factors for improving ovarian function.

10. The method for preparing the capsule containing supramolecular anti-aging factors for improving ovarian function according to claim 9, characterized in that: In the step S9-2, the curing temperature is 0-6°C.

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