Composition containing active peptide and application thereof in improving male function

Abstract

The invention relates to the field of health care products, and particularly discloses a composition containing active peptides and application of the composition in improving male functions. The composition is prepared from 0.5 to 1.0 part of oyster peptide, 1.8 to 2.8 parts of ginseng peptide, 0.5 to 1.0 part of fructus perillae peptide, 1 to 3 parts of cordyceps militaris peptide, 2 to 4 parts of penis et testis cervi peptide, 0.5 to 1.5 parts of cordyceps sinensis extract, 3 to 7 parts of total flavonoids of herba epimedii, 1 to 3 parts of cycloastragenol, 1 to 3 parts of fructus lycii extract, 2 to 6 parts of herba dendrobii extract, 1.5 to 2.5 parts of herba cynomorii extract and 0.05 to 0.15 part of glycine. The composition disclosed by the invention has the effects of tonifying kidney qi, nourishing essence and blood, tonifying Yang, tonifying spleen and replenishing qi, strengthening and consolidating body resistance, resisting fatigue and the like, can also improve the sperm motility and improve the reproductive injury condition of an organism, and has positive influence on the fertility and sexual function of males.

Description

Technical Field

[0001] The present application relates to the field of health care products, and more specifically, to a composition comprising active peptides and its use in improving male function. Background Art

[0002] Erectile dysfunction (ED) refers to the inability of a normal adult male's penis to achieve and maintain an erection sufficient to complete a normal sexual life. According to surveys, more and more men suffer from ED, and the prevalence rate is increasing year by year, especially among the middle-aged and elderly population. The reasons for this phenomenon may be: with the increase of age, the function of the hypothalamus-pituitary gonadal axis gradually declines, and the secretion of gonadotropin decreases significantly, which causes symptoms such as weakened sexual desire and erectile function and decreased muscle strength. When muscle fatigue occurs, the body's sensory information input and motor information output change, and muscle strength and posture control ability decrease, all of which have a certain impact on sexual activity ability. In addition, the increase in the prevalence of cardiovascular and cerebrovascular diseases, hypertension, diabetes, etc., as well as the treatment of these diseases, have damaged the erectile function of the penis to varying degrees. In addition, psychological and social reasons such as tension and anxiety, high life pressure, disharmonious relationship between husband and wife, and unhealthy living habits such as long-term smoking, alcoholism or drug use can cause erectile dysfunction.

[0003] At present, there are many clinical methods for treating ED, such as phosphodiesterase type 5 inhibitors, surgical treatment, vacuum suction and penile prosthesis implantation, etc. However, these treatments are not accepted by most patients due to their serious adverse reactions, and more patients will choose oral medication. However, the drugs currently on the market can also cause certain adverse reactions, such as headaches, nasal congestion, muscle pain, blurred vision, facial flushing, etc., and cannot be taken for a long time. Therefore, it is of great significance to develop a safe and effective food-derived nutritional supplement. Summary of the invention

[0004] In order to solve the above technical problems, the present application provides a composition comprising active peptides, and applies the composition to improve male sexual function.

[0005] In the first aspect, the present application provides a composition comprising an active peptide, which adopts the following technical solution: A composition containing active peptides comprises the following components in parts by weight: 0.5-1.0 parts of oyster peptide, 1.8-2.8 parts of ginseng peptide, 0.5-1.0 parts of perilla seed peptide, 1-3 parts of cordyceps peptide, 2-4 parts of deer whip peptide, 0.5-1.5 parts of cordyceps extract, 3-7 parts of epimedium total flavonoids, 1-3 parts of cycloastragenol, 1-3 parts of wolfberry extract, 2-6 parts of dendrobium extract, 1.5-2.5 parts of cynomorium songaricum extract and 0.05-0.15 parts of glycine.

[0006] By adopting the above technical scheme, the present application rationally combines a variety of tonic medicinal materials, giving full play to the synergistic effect between each other, achieving the effects of tonifying kidney qi, nourishing essence and blood, strengthening yang, tonifying spleen and replenishing qi, strengthening the body and resisting fatigue, and at the same time, it can also improve sperm motility and improve the reproductive damage of the body, thereby having a positive effect on male fertility and sexual function. In addition, the present application also adds a certain amount of glycine to be used in combination with a variety of medicinal materials. Glycine can indirectly regulate male function by affecting the synthesis and release of neurotransmitters (such as dopamine, norepinephrine, etc.), which play an important role in the processes of sexual desire, erection and ejaculation, and also help protect the male reproductive system from damage by oxidative stress and inflammatory response.

[0007] In addition, the present application selects extracts or bioactive peptides of various medicinal materials, fully utilizing the active ingredients in the various medicinal materials, and at the same time making the medicinal materials more easily digestible and absorbed by the human body, significantly improving the effects of the medicinal materials. Moreover, the composition containing active peptides of the present application has few side effects and is a safe and effective food-derived nutritional supplement.

[0008] Preferably, the bioactive peptide composition comprises the following components in parts by weight: 0.8 parts of oyster peptide, 2 parts of ginseng peptide, 0.8 parts of perilla seed peptide, 2 parts of cordyceps peptide, 3 parts of deer whip peptide, 1 part of cordyceps extract, 5 parts of total flavonoids of epimedium, 2 parts of cycloastragenol, 2 parts of wolfberry extract, 4 parts of dendrobium extract, 2 parts of Cynomorium songaricum extract and 0.1 parts of glycine.

[0009] By adopting the above technical solution, the present application further optimizes the dosage ratio between the various components, further exerts the synergistic effect between different medicinal materials, and thus further improves the effect of the composition containing active peptides on improving male sexual function.

[0010] Preferably, the weight ratio of the oyster peptide, ginseng peptide and perilla seed peptide is 1:(3.0-3.5):(0.8-1.2).

[0011] By adopting the above-mentioned technical scheme, the present application adopts a specific proportion of oyster peptide, ginseng peptide and perilla seed peptide mixed and used, which can give full play to the synergistic effect among the three, significantly improve the male's androgen level, gonadotropin level and sperm production ability, and help relieve fatigue, and has a good effect on improving male sexual function.

[0012] Preferably, the oyster peptide is prepared by the following method: a. homogenizing oyster meat and water in a weight ratio of 1:(0.5-2.0) to obtain an oyster homogenate, and then sterilizing the oyster homogenate at a temperature of 120-125° C. for 8-10 minutes to obtain a sterile oyster homogenate; b. After adding lactic acid bacteria to the sterile oyster homogenate, ferment at 36-38°C for 24-25h, the amount of lactic acid bacteria added is 0.02%-2.00% of the mass of the sterile oyster homogenate, and then continue to add yeast, ferment at 40-44°C for 24-25h, the amount of yeast added is 0.02%-2.00% of the mass of the sterile oyster homogenate, and then cook at 100-110°C for 1-7h; c. The result of step b is cooled to room temperature, and the pH value is adjusted to the optimum pH value of the added composite protease, and then the composite protease is added, and enzymolysis is carried out at the optimum temperature of the composite protease for 2-4 hours, and the amount of composite protease added is 1000-5000U / g. After that, the resultant is inactivated, centrifuged at a speed of 4000-8000r / min at room temperature for 5-10min, and the supernatant is spray-dried to obtain oyster peptide.

[0013] By adopting the above technical scheme, the present application obtains oyster peptides containing 3,5-dihydroxy-4-methoxybenzyl alcohol. Compared with ordinary oyster peptides, oyster peptides containing 3,5-dihydroxy-4-methoxybenzyl alcohol have better effects on improving the antioxidant function and metabolic capacity of cells, have more significant anti-fatigue effects, and have more significant effects on improving male sexual function. In addition, the present application controls the amount of lactic acid bacteria added, the amount of yeast added, and the cooking time, so that the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is higher, and the anti-fatigue effect is more significant.

[0014] Preferably, in step b, the amount of lactic acid bacteria added is 0.5%-1.0% of the mass of the sterile oyster homogenate.

[0015] By adopting the above technical solution, the present application further optimizes the amount of lactic acid bacteria added, so that the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in oyster peptides is increased by 0.2%-25.94%, further enhancing the effect of oyster peptides in improving male sexual function.

[0016] Preferably, in step b, the amount of yeast added is 0.5%-1.5% of the mass of the sterile oyster homogenate.

[0017] By adopting the above technical solution, the present application further optimizes the amount of yeast added, so that the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in oyster peptides is increased by 5.38%-28.93%, further enhancing the effect of oyster peptides in improving male sexual function.

[0018] Preferably, in step b, the cooking time is 3-7 hours.

[0019] By adopting the above technical solution, the present application optimizes the cooking time, so that the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in oyster peptides is increased by 2.17%-10.15%, further enhancing the effect of oyster peptides in improving male sexual function.

[0020] Preferably, in step c, the composite protease comprises trypsin and papain, and the enzyme activity ratio of trypsin to papain is (2-3):1.

[0021] By adopting the above technical scheme, the present application adopts a mixture of trypsin and papain to prepare a composite protease. The two proteases have good synergistic effects, which can make the composite protease have an efficient enzymatic hydrolysis effect on oyster meat, so that the product of the prepared oyster peptide is mainly short peptides with a molecular weight of less than 1000Da, accounting for no less than 60%, with high nutritional value and potential biological activity. When other enzymes (such as flavor protease and bromelain) are mixed with trypsin, the proportion of short peptides with a molecular weight of less than 1000Da in the prepared oyster peptides is lower than that of the mixed use of trypsin and papain.

[0022] Preferably, the bioactive peptide composition further comprises 0.3-0.6 parts by weight of sea cucumber peptide and 0.4-0.8 parts by weight of loach peptide.

[0023] By adopting the above-mentioned technical scheme, the present application also adopts a certain amount of sea cucumber peptides and loach peptides mixed with various medicinal materials such as oyster peptides for use, which can give full play to the synergistic effect of the two with medicinal materials such as oyster peptides, further improve the antioxidant capacity of the composition containing active peptides, thereby improving the anti-fatigue effect, and can also enhance the nutritional value of the composition containing active peptides, thereby comprehensively improving the effect of the composition containing active peptides on improving male sexual function.

[0024] Preferably, the sea cucumber peptide is prepared by the following method: The dried sea cucumber body wall is crushed and passed through a 100-mesh sieve to obtain sea cucumber dry powder. The sea cucumber dry powder is mixed with water to obtain a mixed solution with a mass concentration of 0.5%-1.5%, and then the pH value of the mixed solution is adjusted to 5-7, and proteinase K is added at a temperature of 40-60°C, and the amount of proteinase K is 250-350g / U. After enzymolysis for 2-4h, the enzyme is inactivated, cooled, centrifuged, and the supernatant is freeze-dried to obtain sea cucumber peptides.

[0025] By adopting the above-mentioned technical scheme, the present application uses proteinase K to enzymatically hydrolyze sea cucumbers. Compared with those prepared by using other enzymes (such as thermolysin and pepsin), the content of sea cucumber short peptides is higher, the biological activity, stability and specificity are stronger, and the improvement effect of the composition containing active peptides on improving male sexual function is more significant.

[0026] In a second aspect, the present application provides a composition comprising active peptides for use in improving male function.

[0027] In summary, this application has the following beneficial technical effects: 1. The present application rationally combines a variety of tonic medicinal materials with N-glycine, giving full play to the synergistic effect between each other, achieving the effects of invigorating kidney qi, nourishing essence and blood, strengthening yang, invigorating spleen and replenishing qi, strengthening the body and resisting fatigue, and at the same time, it can also improve sperm motility and improve the body's reproductive damage, thereby having a positive impact on male fertility and sexual function; 2. This application uses extracts or bioactive peptides from various medicinal materials, making full use of the active ingredients in each medicinal material, while making the medicinal materials more easily digestible and absorbed by the human body, significantly improving the effects of the medicinal materials; 3. The composition containing active peptides of the present application has little side effects and is a safe and effective food-derived nutritional supplement with extremely broad application prospects. DETAILED DESCRIPTION

[0028] The present application is further described in detail below in combination with preparation examples, embodiments and comparative examples.

[0029] <Material Source> Unless otherwise specified, the raw materials used in this application are all commercially available products, including: Trypsin was purchased from Pangbo Bioengineering Co., Ltd., Nanning, Guangxi, with an enzyme activity of 350,000 U / g; Papain was purchased from Pangbo Bioengineering Co., Ltd., Nanning, Guangxi, with an enzyme activity of 70,000 U / g; Flavor protease, purchased from Pangbo Bioengineering Co., Ltd., Nanning, Guangxi, with an enzyme activity of 70,000 U / g; Bromelain, purchased from Pangbo Bioengineering Co., Ltd., Nanning, Guangxi, with an enzyme activity of 100,000 U / g; Proteinase K was purchased from Shandong Sikoje Biotechnology Co., Ltd. with the product number AA1907, the effective substance concentration was 20 mg / mL, and the enzyme activity was 30 U / mg; Thermolysin was purchased from Shanghai Huzheng Biotechnology Co., Ltd. with a product number of HZM9196 and an enzyme activity of 50 U / mg; Pepsin was purchased from Shanghai Chuangsai Technology Co., Ltd., product number PM11528, enzyme activity was 3000U / mg; Cordyceps sinensis extract, purchased from Shaanxi Snotech Biotechnology Co., Ltd., with an effective content of 98% and a fineness of 100 mesh; Epimedium total flavonoids, purchased from Gansu Yishengxiang Biotechnology Co., Ltd., with an effective content of 40% and a fineness of 200 mesh; Cycloastragenol, purchased from Shaanxi Xiazhou Biotechnology Co., Ltd., with an effective content of 98% and a fineness of 80 mesh; Wolfberry extract, purchased from Lanzhou Waterless Biotechnology Co., Ltd., with an effective content of 98% and a fineness of 120 mesh; Dendrobium officinale extract, purchased from Lanzhou Waterless Biotechnology Co., Ltd., with an effective content of 98% and a fineness of 100 mesh; Cynomorium songaricum extract, purchased from Shaanxi Junhe Biotechnology Co., Ltd., with an effective content of 98% and a fineness of 100 mesh; Ginseng peptides were purchased from Hebei Tuohai Biotechnology Co., Ltd., with an effective content of 99%, short peptides with a molecular weight of less than 500 Da accounting for more than 98%, and a fineness of 120 mesh; Perilla seed peptide, purchased from Shaanxi Snotech Biotechnology Co., Ltd., with a molecular weight of less than 1000Da and a fineness of 100 mesh; Cordyceps militaris peptide, purchased from Shaanxi Sinuo Te Biotechnology Co., Ltd., with a molecular weight of less than 1000 Da and a fineness of 100 mesh; Deer whip peptide, purchased from Lanzhou Waters Biotechnology Co., Ltd., with a molecular weight of less than 1000Da and a fineness of 100 mesh; Glycine, purchased from Xi'an Lavia Biotechnology Co., Ltd., with a fineness of 80 mesh; Loach peptide was purchased from Shanghai Zhongfeng Biotechnology Co., Ltd., with an effective content of 99.2%, a molecular weight of less than 1000Da, and a fineness of 100 mesh.

[0030] <Preparation Example 1.1.1> The oyster peptide is prepared by the following method: a. Shelling fresh oysters, cleaning and crushing them to obtain oyster meat, and then homogenizing them with water at a weight ratio of 1:0.5 to obtain an oyster homogenate, and then sterilizing the oyster homogenate at a temperature of 120° C. for 10 minutes to obtain a sterile oyster homogenate; b. After adding lactic acid bacteria to the sterile oyster homogenate, ferment at 36°C for 25 hours, the amount of lactic acid bacteria added is 1.0% of the mass of the sterile oyster homogenate, and then continue to add yeast, ferment at 40°C for 25 hours, the amount of yeast added is 1.5% of the mass of the sterile oyster homogenate, and then cook at 100°C for 7 hours; c. The obtained product of step b was cooled to room temperature, and the pH value was adjusted to 7.0, and then a composite protease (the ratio of trypsin and papain enzyme activity was 2:1) was added, and the enzymatic hydrolysis was carried out at a temperature of 50°C for 2h, and the amount of composite protease added was 5000U / g, and then the enzyme was inactivated in boiling water for 10min, and centrifuged at room temperature at a speed of 4000r / min for 10min, and the supernatant was spray-dried to obtain oyster peptide. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide was 798.56μg / kg, and the proportion of short peptides with a molecular weight of less than 1000Da in the obtained oyster peptide was 65.61%.

[0031] <Preparation Example 1.1.2> The oyster peptide is prepared by the following method: a. Shelling fresh oysters, cleaning and crushing them to obtain oyster meat, and then homogenizing them with water at a weight ratio of 1:2 to obtain an oyster homogenate, and then sterilizing the oyster homogenate at a temperature of 125° C. for 8 minutes to obtain a sterile oyster homogenate; b. After adding lactic acid bacteria to the sterile oyster homogenate, ferment at 38°C for 24 hours, the amount of lactic acid bacteria added is 1.0% of the mass of the sterile oyster homogenate, and then continue to add yeast, ferment at 44°C for 24 hours, the amount of yeast added is 1.5% of the mass of the sterile oyster homogenate, and then cook at 110°C for 7 hours; c. The obtained product of step b was cooled to room temperature, and the pH value was adjusted to 7.0, and then a composite protease (the ratio of trypsin and papain enzyme activity was 3:1) was added, and the enzymatic hydrolysis was carried out at a temperature of 50°C for 4 hours, and the amount of composite protease added was 1000U / g, and then the enzyme was inactivated in boiling water for 10 minutes, and centrifuged at room temperature at a speed of 8000r / min for 5 minutes, and the supernatant was spray-dried to obtain oyster peptide. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide was 800.24μg / kg, and the proportion of short peptides with a molecular weight of less than 1000Da in the obtained oyster peptide was 60.12%.

[0032] <Preparation Example 1.2.1> The difference from Preparation Example 1.1.1 is that in step b, the amount of lactic acid bacteria added is 0.02% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 632.47 μg / kg.

[0033] <Preparation Example 1.2.2> The difference from Preparation Example 1.1.1 is that in step b, the amount of lactic acid bacteria added is 0.5% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 781.23 μg / kg.

[0034] <Preparation Example 1.2.3> The difference from Preparation Example 1.1.1 is that in step b, the amount of lactic acid bacteria added is 1.5% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 779.65 μg / kg.

[0035] <Preparation Example 1.2.4> The difference from Preparation Example 1.1.1 is that in step b, the amount of lactic acid bacteria added is 2.0% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 731.18 μg / kg.

[0036] <Preparation Example 1.3.1> The difference from Preparation Example 1.1.1 is that in step b, the amount of yeast added is 0.02% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 617.83 μg / kg.

[0037] <Preparation Example 1.3.2> The difference from Preparation Example 1.1.1 is that in step b, the amount of yeast added is 0.5% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 782.33 μg / kg.

[0038] <Preparation Example 1.3.3> The difference from Preparation Example 1.1.1 is that in step b, the amount of yeast added is 1% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 789.66 μg / kg.

[0039] <Preparation Example 1.3.4> The difference from Preparation Example 1.1.1 is that in step b, the amount of yeast added is 2% of the mass of the sterile oyster homogenate, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 742.38 μg / kg.

[0040] <Preparation Example 1.4.1> The difference from Preparation Example 1.1.1 is that in step b, the cooking time is 1 hour, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 723.15 μg / kg.

[0041] <Preparation Example 1.4.2> The difference from Preparation Example 1.1.1 is that in step b, the cooking time is 2 hours, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 729.68 μg / kg.

[0042] <Preparation Example 1.4.3> The difference from Preparation Example 1.1.1 is that in step b, the cooking time is 3 hours, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 745.55 μg / kg.

[0043] <Preparation Example 1.4.4> The difference from Preparation Example 1.1.1 is that in step b, the cooking time is 5 hours, and the rest is the same as Preparation Example 1.1.1. After testing, the content of 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide is 771.27 μg / kg.

[0044] <Preparation Example 1.5.1> The difference from Preparation Example 1.1.1 is that in step c, papain is replaced by flavor protease, and the rest is the same as Preparation Example 1.1.1. After testing, the proportion of short peptides with a molecular weight of less than 1000 Da in the obtained oyster peptides is 50.12%.

[0045] <Preparation Example 1.5.2> The difference from Preparation Example 1.1.1 is that in step c, papain is replaced with bromelain, and the rest is the same as Preparation Example 1.1.1. After testing, the proportion of short peptides with a molecular weight of less than 1000 Da in the obtained oyster peptides is 45.33%.

[0046] <Preparation Example 2.1.1> Sea cucumber peptide is prepared by the following method: The dried sea cucumber body wall is crushed and passed through a 100-mesh sieve to obtain sea cucumber dry powder. The sea cucumber dry powder is mixed with water to obtain a mixed solution with a mass concentration of 0.5%, and then the pH value of the mixed solution is adjusted to 5, and proteinase K is added at a temperature of 40°C, and the amount of proteinase K is 250g / U. After the enzymatic hydrolysis reaction for 4 hours, the enzyme is inactivated in boiling water for 10 minutes, and then cooled to room temperature, centrifuged at a speed of 4000r / min for 15 minutes, and the supernatant is freeze-dried to obtain sea cucumber peptides.

[0047] <Preparation Example 2.1.2> Sea cucumber peptide is prepared by the following method: The dried sea cucumber body wall is crushed and passed through a 100-mesh sieve to obtain sea cucumber dry powder. The sea cucumber dry powder is mixed with water to obtain a mixed solution with a mass concentration of 1.5%, and then the pH value of the mixed solution is adjusted to 7, and proteinase K is added at a temperature of 60°C, and the amount of proteinase K is 350g / U. After the enzymatic hydrolysis reaction for 2h, the enzyme is inactivated in boiling water for 10min, and then cooled to room temperature, centrifuged at a speed of 4000r / min for 15min, and the supernatant is freeze-dried to obtain sea cucumber peptides.

[0048] <Preparation Example 2.1.3> Sea cucumber peptide is prepared by the following method: The dried sea cucumber body wall is crushed and passed through a 100-mesh sieve to obtain sea cucumber dry powder. The sea cucumber dry powder is mixed with water to obtain a mixed solution with a mass concentration of 1%, and then the pH value of the mixed solution is adjusted to 6, and proteinase K is added at a temperature of 50°C, and the amount of proteinase K is 300g / U. After the enzymatic hydrolysis reaction for 3h, the enzyme is inactivated in boiling water for 10min, and then cooled to room temperature, centrifuged at a speed of 4000r / min for 15min, and the supernatant is freeze-dried to obtain sea cucumber peptides.

[0049] <Preparation Example 2.2.1> The difference from Preparation Example 2.1.3 is that proteinase K is replaced by thermolysin, and the rest is the same as Preparation Example 2.1.3.

[0050] <Preparation Example 2.2.2> The difference from Preparation Example 2.1.3 is that proteinase K is replaced by pepsin, and the rest is the same as Preparation Example 2.1.3.

[0051] <Example 1> A method for preparing a composition containing active peptides: 0.5 kg of oyster peptide prepared in Preparation Example 1.1.1, 2.8 kg of ginseng peptide, 0.5 kg of perilla seed peptide, 3 kg of Cordyceps militaris peptide, 2 kg of deer whip peptide, 0.5 kg of Cordyceps sinensis extract, 7 kg of total flavonoids from Epimedium, 1 kg of cycloastragenol, 3 kg of Lycium barbarum extract, 2 kg of Dendrobium officinale extract, 2.5 kg of Cynomorium songaricum extract and 0.15 kg of glycine are mixed evenly, ground for 5 minutes, and passed through a 100-mesh sieve to obtain a composition containing active peptides.

[0052] <Example 2> A method for preparing a composition containing active peptides: 0.5 kg of oyster peptide prepared in Preparation Example 1.1.2, 2.8 kg of ginseng peptide, 0.5 kg of perilla seed peptide, 3 kg of Cordyceps militaris peptide, 2 kg of deer whip peptide, 0.5 kg of Cordyceps sinensis extract, 7 kg of total flavonoids from Epimedium, 1 kg of cycloastragenol, 3 kg of Lycium barbarum extract, 2 kg of Dendrobium officinale extract, 2.5 kg of Cynomorium songaricum extract and 0.15 kg of glycine are mixed evenly, ground for 5 minutes, and passed through a 100-mesh sieve to obtain a composition containing active peptides.

[0053] <Example 3> A method for preparing a composition containing active peptides: 0.8 kg of oyster peptide prepared in Preparation Example 1.1.1, 2 kg of ginseng peptide, 0.8 kg of perilla seed peptide, 2 kg of Cordyceps militaris peptide, 3 kg of deer whip peptide, 1 kg of Cordyceps sinensis extract, 5 kg of total flavonoids from Epimedium, 2 kg of cycloastragenol, 2 kg of Lycium barbarum extract, 4 kg of Dendrobium officinale extract, 2 kg of Cynomorium songaricum extract and 0.1 kg of glycine are mixed evenly, ground for 5 minutes, and passed through a 100-mesh sieve to obtain a composition containing active peptides.

[0054] <Example 4> A method for preparing a composition containing active peptides: 0.8 kg of oyster peptide prepared in Preparation Example 1.1.2, 2 kg of ginseng peptide, 0.8 kg of perilla seed peptide, 2 kg of Cordyceps militaris peptide, 3 kg of deer whip peptide, 1 kg of Cordyceps sinensis extract, 5 kg of total flavonoids from Epimedium, 2 kg of cycloastragenol, 2 kg of Lycium barbarum extract, 4 kg of Dendrobium officinale extract, 2 kg of Cynomorium songaricum extract and 0.1 kg of glycine are mixed evenly, ground for 5 minutes, and passed through a 100-mesh sieve to obtain a composition containing active peptides.

[0055] <Example 5> A method for preparing a composition containing active peptides, which is different from Example 3 in that the weight ratio of oyster peptide, ginseng peptide and perilla seed peptide is 1:3:1.2, and the rest is the same as Example 3, specifically 0.8 kg of oyster peptide, 2.4 kg of ginseng peptide, and 0.96 kg of perilla seed peptide.

[0056] <Example 6> A method for preparing a composition containing active peptides, which is different from Example 3 in that the weight ratio of oyster peptide, ginseng peptide and perilla seed peptide is 1:3.5:0.8, and the rest is the same as Example 3, specifically 0.8 kg of oyster peptide, 2.8 kg of ginseng peptide, and 0.64 kg of perilla seed peptide.

[0057] <Example 7> A method for preparing a composition containing active peptides, which is different from Example 3 in that the weight ratio of oyster peptide, ginseng peptide and perilla seed peptide is 1:3.25:1, and the rest is the same as Example 3, specifically 0.8 kg of oyster peptide, 2.6 kg of ginseng peptide, and 0.8 kg of perilla seed peptide.

[0058] <Example 8-21> A method for preparing a composition containing active peptides, which is different from Example 3 in that the oyster peptides prepared in Preparation Example 1.1.1 are replaced by the oyster peptides prepared in Preparation Examples 1.2.1-1.5.2 respectively, and the rest is the same as Example 3.

[0059] <Example 22> A method for preparing a composition containing active peptides, which is different from Example 3 in that: 0.3 kg of sea cucumber peptide prepared in Preparation Example 2.1.1 and 0.8 kg of loach peptide are added and mixed with oyster peptide and other components, and the rest is the same as Example 3.

[0060] <Example 23> A method for preparing a composition containing active peptides, which is different from Example 3 in that: 0.6 kg of sea cucumber peptide prepared in Preparation Example 2.1.2 and 0.4 kg of loach peptide are added and mixed with oyster peptide and other components, and the rest is the same as Example 3.

[0061] <Example 24> A method for preparing a composition containing active peptides, which is different from Example 3 in that: 0.45 kg of sea cucumber peptide prepared in Preparation Example 2.1.3 and 0.6 kg of loach peptide are added and mixed with oyster peptide and other components, and the rest is the same as Example 3.

[0062] <Examples 25-26> A method for preparing a composition containing active peptides, which is different from Example 24 in that the sea cucumber peptides prepared in Preparation Example 2.1.3 are replaced by the sea cucumber peptides prepared in Preparation Examples 2.2.1-2.2.2 respectively, and the rest is the same as Example 24.

[0063] <Comparative Example 1> The difference from Example 3 is that the oyster peptide is prepared by the following method: a. Shelling fresh oysters, cleaning and crushing to obtain oyster meat, and then homogenizing with water at a weight ratio of 1:0.5 to obtain oyster homogenate; b. Adjust the pH value of the oyster homogenate to 7.0, then add a composite protease (the ratio of trypsin and papain is 2:1), and enzymolyze at 50°C for 2h. The amount of composite protease added is 5000U / g, then inactivate the enzyme in boiling water for 10min, centrifuge at 4000r / min for 10min at room temperature, take the supernatant for spray drying, and obtain oyster peptide. After testing, the 3,5-dihydroxy-4-methoxybenzyl alcohol in the obtained oyster peptide was lower than the detection limit of 0.05μg / mL.

[0064] <Comparative Example 2> The difference from Example 3 is that ginseng peptide is not added, the amount of oyster peptide is adjusted to 1.9 kg, the amount of perilla seed peptide is adjusted to 1.9 kg, and the rest is the same as Example 3.

[0065] <Comparative Example 3> The difference from Example 3 is that no perilla seed peptide is added, the amount of oyster peptide is adjusted to 0.75 kg, the amount of ginseng peptide is adjusted to 3.05 kg, and the rest is the same as Example 3.

[0066] <Comparative Example 4> The difference from Example 3 is that the perilla seed peptide is replaced by soybean peptide, and the rest is the same as Example 3. The soybean peptide is purchased from Shaanxi Snotech Biotechnology Co., Ltd., and the short peptides with a molecular weight of less than 500Da account for more than 98%, and the fineness is 100 mesh.

[0067] <Comparative Example 5> The difference from Example 3 is that ginseng peptide is replaced by collagen peptide, and the rest is the same as Example 3. The collagen peptide is purchased from Shaanxi Rankang Biotechnology Co., Ltd., with a molecular weight of less than 1000Da and a fineness of 100 mesh.

[0068] <Performance Testing> 1. Anti-fatigue effect detection: 320 male ICR experimental mice were divided into a blank control group and the remaining 31 groups, each group had 10 ICR mice, wherein the mice in the blank control group were orally gavaged with water for 28 consecutive days, and the mice in the remaining 31 groups were orally gavaged with drugs at a dose of 1g / kg for 28 consecutive days, and anti-fatigue experiments were carried out 0.5 hours after the last administration, wherein the drugs used in the 31 groups corresponded to the products prepared in Examples 1-26 and Comparative Examples 1-5, respectively. The anti-fatigue effect experiment was specifically as follows: 0.5 hours after the last administration, 5 mice were randomly selected from each group, 5% of the body weight of lead blocks were tied to the roots of the mice, and they were placed in a swimming box to swim in turn, the water depth of the swimming box was 30cm, the water temperature was (25±1.0)℃, and the time from the start of the mouse's swimming to the time when the mouse sank to the bottom of the box and could not float to the surface of the water was recorded as the weighted swimming time of the mouse.

[0069] 2. Detection of the effect of improving male sexual function: This application reflects the effect of improving male sexual function by detecting the production of androgens in testicular interstitial cells, specifically: ① TM3 cell culture: Use DMEM / F12 (1:1) culture medium containing 5% HS and 2.5% FBS, and culture in an incubator at 37°C, 5% CO2 and sufficient saturated humidity. According to the actual situation of the cells, change the medium every 2-3 days. When the cells are fused to 80% of the culture dish, discard the old culture medium in the culture flask, rinse the cells with PBS buffer 2-3 times, add an appropriate amount of 0.05% trypsin digestion solution rewarmed at room temperature, and culture according to 1:3. After 3 subcultures, take cells in the logarithmic phase and in good growth state for experiments; ② TM3 survival rate detection: After digesting the cells in the logarithmic growth phase, use DMEM / F12 complete culture medium (containing 10% FBS and 1% penicillin-streptomycin The cell density was adjusted to 105 cells / mL by adding a cell suspension (100 μL) and the cells were plated in a 96-well plate. Each well was incubated in a CO2 incubator for 24 hours. After the cell monolayer adhered to the wall, the DMEM / F12 incomplete medium was replaced. The sample solutions with a concentration of 400 μg / mL were added respectively to obtain different groups. The sample solutions of different groups corresponded to the products obtained in Examples 1-26 and Comparative Examples 1-5, respectively. The cells were cultured for 24 hours, and then the cell survival rate was detected by the MTT method. The calculation formula was: survival rate (%) = OD sample / OD control × 100%; ③ NO (nitrate reductase method), testosterone (competitive ELISA method), dihydrotestosterone (competitive ELISA method) and intracellular SOD (WST-8 method), MDA (thiobarbituric acid method), and cGMP production (competitive ELISA method) in the cell culture supernatant of different groups were detected respectively.

[0070] Table 1 Weighted swimming time of mice in the anti-fatigue effect experiment Table 2 Test results of improving male sexual function Table 2 It can be seen from Table 1 and Table 2 that the weighted swimming time of mice in the corresponding groups of Examples 1-4 was longer than that of the control group, and in TM3 cells, the contents of NO, testosterone, dihydrotestosterone, SOD and cGMP were higher than those of the control group, and the content of MDA was lower than that of the control group. Moreover, the survival rate of TM3 cells was not less than 120% after detection. Testosterone can promote spermatogenesis, maturation and arginine production; NO is a non-adrenal non-cholinergic neurotransmitter, which is the key substance that causes penile smooth muscle relaxation and blood to enter the corpus cavernosum to fill and erect; dihydrotestosterone can be converted into 17β-estradiol under the action of aromatase, which is widely distributed in the blood throughout the body and can bind to the androgen receptors of spermatogenic cells to promote spermatogenesis. Its main functions are to maintain spermatogenesis, normal sexual desire and stimulate the development of reproductive organs. It is an important male hormone; SOD enzyme activity and MDA content are two commonly used indicators for evaluating the body's antioxidant capacity. The SOD enzyme in testicular cells can maintain the normal oxidative environment of cells and regulate spermatogenesis in spermatogenic cells; penile erection refers to the phenomenon that the corpus cavernosum of the penis is rapidly filled with blood due to sexual stimulation, and the penis quickly becomes filled with blood, hardens and extends. The parasympathetic nerve center activates soluble adenylate cyclase by releasing soluble neurotransmitter NO, and the intracellular GTP is converted into cGMP. As the second messenger in the cell, cGMP can make the Ca in the cell 2+ The concentration of intracellular Ca 2+ The corpus cavernosum is reduced, the corpus cavernosum smooth muscle relaxes, the intracavernosal pressure increases, and at the same time, the subalbuminae venules are passively compressed and occluded, resulting in erection. Therefore, the test data show that the composition containing active peptides prepared in Examples 1-4 of the present application has a significant effect on fighting fatigue and improving male sexual function, and when the ratio between the drugs is further optimized, the effect of fighting fatigue and improving male sexual function is more prominent.

[0071] The weighted swimming time of the mice in the corresponding groups of Examples 5-7 was higher than that in Example 3, and in TM3 cells, the contents of NO, testosterone, dihydrotestosterone, SOD and cGMP were higher than those in Example 3, and the MDA content was lower than that in Example 3. This indicates that when the ratio between oyster peptide, ginseng peptide and perilla seed peptide is further optimized, the synergistic effect between the three can be further exerted, thereby further improving the improvement effect of the composition containing active peptides in anti-fatigue and male sexual function.

[0072] The difference between Example 8-21 and Example 3 is that the preparation method of oyster peptide is different. It can be seen from Table 1 and Table 2 that when the content of 3,5-dihydroxy-4-methoxybenzyl alcohol and the proportion of short peptides in oyster peptides are further optimized, the effect of oyster peptides themselves can be further promoted, thereby further improving the improvement effect of the composition containing active peptides in anti-fatigue and male sexual function.

[0073] The weighted swimming time of mice in the corresponding groups of Examples 22-24 was higher than that in Example 3, and in TM3 cells, the contents of NO, testosterone, dihydrotestosterone, SOD and cGMP were higher than those in Example 3, and the MDA content was lower than that in Example 3. This indicates that when sea cucumber peptides and loach peptides are further added, the synergistic effect between the two and components such as oyster peptides can be fully exerted, thereby further improving the anti-fatigue and male sexual function improvement effects of the composition containing active peptides.

[0074] The difference between Examples 25-26 and Example 24 is that the preparation methods of sea cucumber peptides are different. It can be seen from Tables 1 and 2 that when the proportion of short peptides in sea cucumber peptides is further optimized, the effects of sea cucumber peptides themselves can be further promoted, thereby further improving the improvement effect of the composition containing active peptides in anti-fatigue and male sexual function.

[0075] The weighted swimming time of the mice in the corresponding group of Comparative Example 1 was lower than that in Example 3, and in TM3 cells, the contents of NO, testosterone, dihydrotestosterone, SOD and cGMP were lower than those in Example 3, and the MDA content was higher than that in Example 3. This shows that the oyster peptide containing 3,5-dihydroxy-4-methoxybenzyl alcohol used in this application can further improve the improvement effect of the composition containing active peptides in anti-fatigue and male sexual function.

[0076] The weighted swimming time of mice in the corresponding groups of Comparative Examples 2-5 was lower than that in Example 3, and in TM3 cells, the contents of NO, testosterone, dihydrotestosterone, SOD and cGMP were all lower than those in Example 3, and the MDA content was higher than that in Example 3. This shows that the present application gives full play to the synergistic effect between oyster peptides, perilla seed peptides and ginseng peptides, so that the composition containing active peptides of the present application has a significant effect on anti-fatigue and improving male sexual function.

[0077] The embodiments of this specific implementation method are all preferred embodiments of the present application, and are not intended to limit the protection scope of the present application. Therefore, all equivalent changes made based on the structure, shape, and principle of the present application should be included in the protection scope of the present application.

Claims

1. A composition comprising an active peptide, characterized in that: The invention comprises the following components in parts by weight: 0.5-1.0 parts of oyster peptide, 1.8-2.8 parts of ginseng peptide, 0.5-1.0 parts of perilla seed peptide, 1-3 parts of cordyceps peptide, 2-4 parts of deer whip peptide, 0.5-1.5 parts of cordyceps sinensis extract, 3-7 parts of total flavonoids of epimedium, 1-3 parts of cycloastragenol, 1-3 parts of wolfberry extract, 2-6 parts of dendrobium extract, 1.5-2.5 parts of cynomorium songaricum extract and 0.05-0.15 parts of glycine.

2. A composition comprising an active peptide according to claim 1, characterized in that: The invention comprises the following components in parts by weight: 0.8 parts of oyster peptide, 2 parts of ginseng peptide, 0.8 parts of perilla seed peptide, 2 parts of cordyceps peptide, 3 parts of deer whip peptide, 1 part of cordyceps extract, 5 parts of total flavonoids of epimedium, 2 parts of cycloastragenol, 2 parts of wolfberry extract, 4 parts of dendrobium extract, 2 parts of cynomorium songaricum extract and 0.1 parts of glycine.

3. A composition comprising an active peptide according to claim 1, characterized in that: The weight ratio of the oyster peptide, ginseng peptide and perilla seed peptide is 1:(3.0-3.5):(0.8-1.2).

4. A composition comprising an active peptide according to any one of claims 1 to 3, characterized in that: The oyster peptide is prepared by the following method: a. homogenizing oyster meat and water in a weight ratio of 1:(0.5-2.0) to obtain an oyster homogenate, and then sterilizing the oyster homogenate at a temperature of 120-125°C for 8-10 minutes to obtain a sterile oyster homogenate; b. After adding lactic acid bacteria to the sterile oyster homogenate, ferment at 36-38°C for 24-25h, the amount of lactic acid bacteria added is 0.02%-2.00% of the mass of the sterile oyster homogenate, and then continue to add yeast, ferment at 40-44°C for 24-25h, the amount of yeast added is 0.02%-2.00% of the mass of the sterile oyster homogenate, and then cook at 100-110°C for 1-7h; c. The result of step b is cooled to room temperature, and the pH value is adjusted to the optimum pH value of the added composite protease, and then the composite protease is added, and enzymolysis is carried out at the optimum temperature of the composite protease for 2-4 hours, and the amount of composite protease added is 1000-5000U / g. After that, the resultant is inactivated, centrifuged at a speed of 4000-8000r / min at room temperature for 5-10min, and the supernatant is spray-dried to obtain oyster peptide.

5. A composition comprising an active peptide according to claim 4, characterized in that: In the step b, the amount of lactic acid bacteria added is 0.5%-1.0% of the mass of the sterile oyster homogenate.

6. A composition comprising an active peptide according to claim 4, characterized in that: In the step b, the amount of yeast added is 0.5%-1.5% of the mass of the sterile oyster homogenate.

7. A composition comprising an active peptide according to claim 4, characterized in that: In the step b, the cooking time is 3-7 hours.

8. A composition comprising an active peptide according to claim 1, characterized in that: In the step c, the composite protease includes trypsin and papain, and the enzyme activity ratio of the trypsin and papain is (2-3):

1.

9. A composition comprising an active peptide according to claim 1, characterized in that: The bioactive peptide composition further comprises 0.3-0.6 parts by weight of sea cucumber peptide and 0.4-0.8 parts by weight of loach peptide.

10. Use of the composition comprising active peptides as claimed in claim 1 in improving male function.

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