Composition for relaxing bowel, reducing blood sugar, losing weight, resisting aging and promoting sleep

Abstract

The invention provides a composition for relaxing bowel, reducing blood sugar, losing weight, resisting aging and promoting sleep, which can be used in the fields of biology, medicine, food, health care products and the like. In some embodiments, the composition comprises a proper amount of water (optional), a recombinant collagen-like protein (FLM1.6 PHPV fusion factor, fusion factor for short), gamma-aminobutyric acid, chia seeds, walnut kernels, carrots and cassia seeds, and the amino acid sequence of the recombinant collagen-like protein is as shown in SEQ ID NO: 1. The invention also provides the recombinant collagen-like protein, and a medicine, a beverage, powder and a tablet candy containing the composition. Optionally, the composition is a medicinal and edible dual-purpose composition. The composition and the protein have unexpected effects of relaxing bowel, reducing blood sugar, losing weight, resisting aging and promoting sleep to a certain extent. Good market prospects are realized.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and food health products, and specifically relates to a composition for moistening the intestines and facilitating bowel movements, lowering blood sugar and reducing weight, and resisting aging and promoting sleep. Background Art

[0002] Lactoferrin is an iron-binding lactose protein that promotes the growth of specific probiotic strains and can affect the growth and diversification of the intestinal microbiota, including (i) enhancing the permeability of the epithelial cell monolayer and (ii) providing signals to balance the anti-inflammatory and pro-inflammatory responses that lead to intestinal homeostasis. The effects of lactoferrin on the intestinal barrier are mainly reflected in the following aspects: repairing the damaged intestinal physical barrier by promoting intestinal epithelial cell proliferation, increasing the expression of tight junction proteins, and improving intestinal tissue morphology; protecting the intestinal immune barrier by activating intestinal immune cells and regulating the secretion of cytokines; protecting the intestinal biological barrier by positively regulating the distribution of intestinal flora; and protecting the intestinal chemical barrier by enhancing mucin expression and regulating the secretion of antimicrobial peptides. In general, lactoferrin has multiple biological functions, including antimicrobial activity, anti-inflammatory activity, intestinal barrier protection, and immune cell regulation, and is involved in maintaining intestinal mucosal immune homeostasis. It has been reported that lactoferrin alleviates cognitive impairment caused by Western diet through the microbiome-gut-brain axis.

[0003] Intestinal collagen plays an important role in intestinal health, mainly in the following aspects: strengthening the intestinal barrier, collagen helps to support and strengthen the protective layer of the digestive tract to prevent the occurrence of leaky gut syndrome; promoting tissue repair, when the intestine is injured, the accumulation of collagen can stimulate cell regeneration and help repair damaged tissue; improving the intestinal lining, collagen is rich in glycine, which helps to rebuild the intestinal lining tissue and reduce intestinal inflammation; regulating intestinal function, the amino acids in collagen are believed to help regulate gastric acid production and prevent gastric ulcers. In summary, intestinal collagen is essential for maintaining healthy intestinal function.

[0004] Interestingly, collagen peptides have been reported to exhibit probiotic properties through interaction with the gut microbiota, which can alleviate symptoms associated with metabolic diseases through the production of short-chain and branched-chain fatty acids.

[0005] Gamma-aminobutyric acid (GABA) is an important inhibitory neurotransmitter in the central nervous system. It has been confirmed that GABA is safe for consumption and has physiological effects such as improving sleep quality and lowering blood pressure. Modern pharmacological studies have shown that the main components of Cassia seed include enquinones, phenylpropionazoles, fatty acids, glycosides, etc., which have the effects of protecting the liver, improving eyesight, protecting nerves, and helping sleep. The GI value of chia seeds is less than 55, which is a low GI food; the GI value refers to the value of how fast blood sugar rises after food enters the body. Foods with low GI values ​​will raise blood sugar more slowly, so they help prevent the body's blood sugar waves from being too high to maintain cardiovascular health. The latest research shows that carrots have the potential to improve blood sugar regulation and promote healthier intestinal flora, or can provide a natural and potentially side-effect-free auxiliary diet therapy for diabetic patients. It is reported that consuming 2-3 walnut kernels a day can reduce total blood cholesterol and low-density cholesterol. Summary of the invention

[0006] In one aspect of the present invention, a composition for moistening the intestines and promoting bowel movements, lowering blood sugar and reducing weight, and resisting aging and promoting sleep is provided, the composition comprising a protein with an amino acid sequence as shown in SEQ ID NO: 1. The applicant named the protein with an amino acid sequence as shown in SEQ ID NO: 1 as FLM1.6 PHPV fusion factor.

[0007] In another aspect of the present invention, a composition for moisturizing the intestines and promoting bowel movements, lowering blood sugar and losing weight, and resisting aging and promoting sleep is provided, the composition comprising recombinant collagen, γ-aminobutyric acid, chia seeds, walnut kernels, carrots, and cassia seeds, and the amino acid sequence of the recombinant collagen is shown in SEQ ID NO: 1.

[0008] Walnut kernels contain high levels of plant fat and protein, including 70% fat and 17%-27% protein. Walnut kernels contain not only calcium, iron, phosphorus and other elements, but also zinc, manganese, chromium and other essential minerals for the human body. Multiple vitamins, proteins and minerals are all nutrients that are beneficial to a healthy appearance. The linoleic acid in the plant fat rich in walnut kernels is regarded as a beauty agent that nourishes and moisturizes the skin, making it smooth and elastic.

[0009] Optionally, the composition is a medicinal and edible composition.

[0010] In some specific embodiments of the present invention, the composition comprises 0.05-0.2 parts by weight of recombinant collagen, 0.05-0.2 parts by weight of γ-aminobutyric acid, 1.5-2.5 parts by weight of chia seeds, 0.5-2 parts by weight of walnut kernels, 0.5-2 parts by weight of carrots, and 0.5-2 parts by weight of cassia seeds.

[0011] In some specific embodiments of the present invention, the composition comprises 0.1 parts by weight of recombinant collagen, 0.1 parts by weight of γ-aminobutyric acid, 2 parts by weight of chia seeds, 1 part by weight of walnut kernels, 1 part by weight of carrots, and 1 part by weight of cassia seeds.

[0012] In some specific embodiments of the present invention, the chia seeds are chia seed powder. In some specific embodiments of the present invention, the walnut kernels are walnut kernel powder. In some specific embodiments of the present invention, the carrots are carrot powder. In some specific embodiments of the present invention, the cassia seeds are cassia seeds powder.

[0013] In some specific embodiments of the present invention, the Cassia seeds are mature seeds of Cassia.

[0014] In some specific embodiments of the present invention, the composition further comprises water. In some specific embodiments of the present invention, the composition comprises an appropriate amount of water.

[0015] In some specific embodiments of the present invention, the composition further comprises 20-30 parts by weight of water. In some specific embodiments of the present invention, the composition further comprises 25 parts by weight of water.

[0016] In some specific embodiments of the present invention, the preparation method of the composition is as described in Example 2. In some specific embodiments of the present invention, the preparation method of the composition is as described in Example 3.

[0017] In some specific embodiments of the present invention, the composition comprises 0.1 parts by weight of recombinant collagen, 0.1 parts by weight of γ-aminobutyric acid, 2 parts by weight of chia seeds, 1 part by weight of walnut kernels, 1 part by weight of carrots, 1 part by weight of cassia seeds, and 20-30 parts by weight of water.

[0018] In some specific embodiments of the present invention, the composition comprises 0.1 parts by weight of recombinant collagen, 0.1 parts by weight of γ-aminobutyric acid, 2 parts by weight of chia seed powder, 1 part by weight of walnut kernel powder, 1 part by weight of carrot powder, 1 part by weight of cassia seed powder, and 20-30 parts by weight of water.

[0019] In some specific embodiments of the present invention, the composition comprises 0.1 parts by weight of recombinant collagen, 0.1 parts by weight of γ-aminobutyric acid, 2 parts by weight of chia seed powder, 1 part by weight of walnut kernel powder, 1 part by weight of carrot powder, 1 part by weight of cassia seed powder, and 25 parts by weight of water.

[0020] In some specific embodiments of the present invention, the composition is a composition for moistening the intestines and promoting bowel movements, lowering blood sugar and losing weight, and resisting aging and promoting sleep.

[0021] In another aspect of the present invention, a protein is provided for moistening the intestines and promoting bowel movements, lowering blood sugar and reducing weight, and resisting aging and promoting sleep, wherein the amino acid sequence of the protein is shown in SEQ ID NO: 1 or 3.

[0022] In some specific embodiments of the present invention, the amino acid sequence of the protein is shown in SEQ ID NO: 1. In some specific embodiments of the present invention, the amino acid sequence of the protein is shown in SEQ ID NO: 3.

[0023] In another aspect of the present invention, a nucleic acid is provided, wherein the nucleic acid encodes the protein described in the present invention.

[0024] In another aspect of the present invention, a medicine for moistening the intestines and promoting bowel movements, lowering blood sugar and reducing weight, and resisting aging and promoting sleep is provided, wherein the medicine comprises the composition as described in the present invention or the protein as described in the present invention.

[0025] In some specific embodiments of the present invention, the medicament comprises the composition as described in the present invention. In some specific embodiments of the present invention, the medicament comprises the protein as described in the present invention.

[0026] In another aspect of the present invention, a beverage is provided which has the functions of moistening the intestines and promoting bowel movements, lowering blood sugar and reducing weight, and resisting aging and promoting sleep. The beverage comprises the composition as described in the present invention or the protein as described in the present invention.

[0027] In some specific embodiments of the present invention, the beverage comprises the composition as described in the present invention. In some specific embodiments of the present invention, the beverage comprises the protein as described in the present invention.

[0028] In another aspect of the present invention, a food having the functions of moistening the intestines and promoting bowel movements, lowering blood sugar and reducing weight, resisting aging and promoting sleep is provided, wherein the food comprises the composition as described in the present invention or the protein as described in the present invention.

[0029] In some specific embodiments of the present invention, the food comprises the composition as described in the present invention. In some specific embodiments of the present invention, the food comprises the protein as described in the present invention.

[0030] In another aspect of the present invention, a powder for moistening the intestines and promoting bowel movements, lowering blood sugar and reducing weight, and resisting aging and promoting sleep is provided, wherein the powder comprises the composition as described in the present invention or the protein as described in the present invention.

[0031] In another aspect of the present invention, a compressed candy is provided, wherein the compressed candy comprises the composition according to the present invention or the protein according to the present invention.

[0032] In another aspect of the present invention, there is provided use of the composition of the present invention or the protein of the present invention in moistening the intestines and relieving constipation, lowering blood sugar and losing weight, and resisting aging and promoting sleep.

[0033] In another aspect of the present invention, there is provided use of the composition of the present invention or the protein of the present invention in the preparation of a drug for moistening the intestines and facilitating bowel movements, lowering blood sugar and reducing weight, and resisting aging and promoting sleep.

[0034] In some specific embodiments of the present invention, there is provided use of the composition of the present invention in the preparation of medicaments for laxative, hypoglycemic and anti-aging effects.

[0035] In some specific embodiments of the present invention, there is provided the use of the composition of the present invention in the preparation of a laxative drug. In some specific embodiments of the present invention, there is provided the use of the composition of the present invention in the preparation of a drug for lowering blood sugar. In some specific embodiments of the present invention, there is provided the use of the composition of the present invention in the preparation of a drug for anti-aging.

[0036] In some specific embodiments of the present invention, there is provided use of the protein of the present invention in the preparation of medicaments for laxative, hypoglycemic and anti-aging effects.

[0037] In some specific embodiments of the present invention, there is provided the use of the protein of the present invention in the preparation of a laxative drug. In some specific embodiments of the present invention, there is provided the use of the protein of the present invention in the preparation of a drug for lowering blood sugar. In some specific embodiments of the present invention, there is provided the use of the protein of the present invention in the preparation of a drug for anti-aging.

[0038] The composition and protein of the present invention have strong laxative effect, as well as certain blood sugar lowering, weight loss, anti-aging and sleep promoting effects. It has a good market prospect. The food, beverage and health care product of the present invention have certain laxative effect, blood sugar lowering, weight loss, anti-aging and sleep promoting effects. Unexpected technical effects have been achieved.

[0039] The beverage of the present invention may include appropriate amounts of water, recombinant collagen, gamma-aminobutyric acid, chia seeds, walnut kernels, carrots, and cassia seeds. DETAILED DESCRIPTION

[0040] The present disclosure is further illustrated by way of examples below, but the present disclosure is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are selected according to conventional methods and conditions, or according to the product instructions. Unless otherwise specified, the reagents and raw materials used in the present disclosure are all commercially available.

[0041] Example 1. Design and preparation of recombinant collagen-like protein (fusion factor) Based on years of in-depth research in the field of collagen, the applicant has designed a recombinant collagen-like protein (named FLM1.6 PHPV fusion factor, referred to as fusion factor), the sequence of which is shown in SEQ ID NO: 1. The protein structure is PEP-1-linker-collagen-like sequence-linker-lactoferrin-like sequence-His tag. The fusion factor PHPV is prominent, contains sequence-optimized collagen fragments, which can repair intestinal mucosal damage; contains sequence-optimized biological defense protein fragments (chimera of bovine and human lactoferrin conservative sequences) which can improve intestinal microecology.

[0042] The nucleic acid fragment encoding the recombinant collagen-like protein (fusion factor) (codon optimized for E. coli) was cloned into the BlpI and XbaI restriction sites of the pET28a(+) vector. The reaction solution was transformed into Stbl3 competent cells and cultured for 15 hours, after which positive clones were selected for sequencing for identification. The positive clones with correct sequencing were then cultured and the expression vector plasmid of the recombinant collagen-like protein was extracted (SEQ ID NO: 2).

[0043] The obtained vector plasmid was transformed into BL21(DE3) E. coli competent cells (NEB, C2527I). Cultured in LB medium containing kanamycin at 37°C for 16h. IPTG was added to a concentration of 0.5 mM and induced at 16°C for 19h. The cells were collected by centrifugation, and the lysate was added and ultrasonically broken. The supernatant containing recombinant collagen was obtained by centrifugation again. Filtered with a 0.45µm filter membrane, purified with a Ni-NTA affinity chromatography column, and ion chromatography, the eluate was freeze-dried to obtain a recombinant collagen with a purity of 96.1% (SEQ ID NO: 1). It only showed a single band in SDS-PAGE.

[0044] The same method was used to prepare a collagen-like protein (SEQ ID NO: 3). The protein structure is PEP-1-linker-collagen sequence-linker-lactoferrin sequence-His tag.

[0045] Example 2. Preparation of the composition Take fresh, ripe carrots, peel them, and cut them into small pieces with a length, width, and height of 2-3 mm. Put the carrot pieces into a hot air dryer at 60 degrees Celsius and dry them for about 6-10 hours until the moisture content drops to 6%-10%. Use a pulverizer to pulverize the dried carrot pieces to an average particle size of about 100 microns to obtain carrot powder.

[0046] Take dry and clean chia seeds and use a grinder to grind them into an average particle size of about 100 microns to obtain chia seed powder.

[0047] Take dry and clean walnut kernels, use liquid nitrogen to freeze and crush them to an average particle size of about 200 microns, and obtain walnut kernel powder.

[0048] Take dry and clean Cassia seeds (mature seeds of Cassia), and use a grinder to grind them into an average particle size of about 80 microns to obtain Cassia seed powder.

[0049] Take 100 mg of recombinant collagen (fusion factor), 100 mg of γ-aminobutyric acid, 2 g of chia seed powder, 1 g of walnut kernel powder, 1 g of carrot powder, and 1 g of cassia seed powder, mix them evenly to obtain a solid powder, which is the composition CTS-1.

[0050] Take 100 mg of collagen-like protein (SEQ ID NO: 3), 100 mg of γ-aminobutyric acid, 2 g of chia seed powder, 1 g of walnut kernel powder, 1 g of carrot powder, and 1 g of cassia seed powder, mix them evenly to obtain a solid powder, which is the composition CTS-2.

[0051] Take 100 mg of γ-aminobutyric acid, 2 g of chia seed powder, 1 g of walnut kernel powder, 1 g of carrot powder, and 1 g of cassia seed powder, mix them evenly to obtain a solid powder, which is the composition CTS-3.

[0052] Example 3. Laxative function test Adult C57BL / 6 mice (female), weighing 19.7-20.3g, were randomly divided into groups, 10 in each group. The modeling drug compound diphenoxylate was given by oral gavage to obtain a constipation model of mice. The time of the first black stool, the number of stools within 5 hours and the stool weight were measured by gavage with ink to reflect the defecation condition of the model mice.

[0053] Each composition (CTS-n, n is a number) prepared as described in Example 2 was stirred in sterile water at room temperature for 5 minutes to obtain a uniform solid-liquid mixture (CTS-nL, n is a number corresponding to the number n of CTS-n) with a concentration of 200 mg composition / mL.

[0054] The recombinant collagen (fusion factor) prepared by the method described in the previous example was stirred in sterile water at room temperature for 5 minutes to obtain a uniform suspension with a concentration of 20 mg / mL.

[0055] The mice in group 1 were gavaged with the solid-liquid mixture CTS-1-L, the mice in group 2 were gavaged with the composition CTS-2-L, the mice in group 3 were gavaged with the composition CTS-3-L, and the mice in group 4 were gavaged with the recombinant collagen suspension; gavage was once a day. The volume administered was 10 mL / kg body weight. The mice in the normal control group and the model group were given 10 mL / kg body weight of sterile water.

[0056] After continuous gavage for 8 days, the mice in each group were fasted but not watered for 18 hours. The normal control group was given normal saline, and the mice in groups 1-4 and the model group were gavaged with a suspension of compound diphenoxylate tablets in water (25 mg / mL of diphenoxylate concentration) at 25 mg / kg body weight. After 30 minutes, the mice in each group were gavaged with an equal amount of ink. From the beginning of the ink gavage, the discharge time of the first black stool of each mouse and the weight of black stool in a specific time period were recorded. As shown in Table 1, the average value of mice in the group was taken for each data. The weight of black stool discharged within 12 hours in groups 1 and 4 was significantly higher than that in the model group, group 2 and group 3, respectively, with statistically significant differences (P<0.05); in addition, the discharge time of the first black stool in groups 1 and 4 was significantly shorter than that in the model group, group 2 and group 3 (P<0.05). It shows that both composition 1 and recombinant collagen have strong laxative functions.

[0057] Table 1. Laxative function test results Example 4. Blood sugar lowering function test Male healthy Kunming mice (weight 19.0-22.3 g) were taken and fed normally for 3 days, and then fasted but not watered for 15 hours. Ten mice were randomly selected as the normal control group; the other mice were used for modeling, and each mouse was intraperitoneally injected with 160 mg / kg streptozotocin. The fasting blood glucose of the mice was measured 48 hours later, and the mice with blood glucose concentration higher than 11.1 mmol / L were considered to have successfully established the diabetes model.

[0058] Then the model mice were randomly divided into groups, with 10 mice in each group. The test group was gavaged once a day with the test substance, i.e., recombinant collagen (SEQ ID NO: 1) and collagen (SEQ ID NO: 3), respectively, at 50 mg / mL, stirred in water for 1 hour and dispersed evenly before gavage, with the volume of 10 mL / kg body weight.

[0059] Another group of mice was gavaged once daily with the solid-liquid mixture CTS-1-L (200 mg of the composition CTS-1 / mL) prepared as in the previous example, at a volume of 10 mL / kg body weight.

[0060] The normal control group and the model group were gavaged with an equal volume of normal saline daily for 12 consecutive days. After fasting for 12 hours on the last day of gavage, fasting blood glucose was measured (the results were expressed as mean values). The results are shown in Table 2.

[0061] The fasting blood glucose of mice in the recombinant collagen group after administration was significantly lower than that in the model group, with a statistically significant difference (P<0.05), and was also significantly lower than that in the collagen group (P<0.05). The fasting blood glucose of mice in the solid-liquid mixture CTS-1-L group after administration was also significantly lower than that in the model group (P<0.05), indicating that the recombinant collagen and the solid-liquid mixture CTS-1-L have a certain blood sugar-lowering function, thus achieving an unexpected technical effect.

[0062] Table 2. Fasting blood glucose of mice Example 5. Anti-aging function test Male C57BL / 6 mice, 8 weeks old and weighing 19.1-21.2 g, were randomly divided into groups, with 10 animals in each group.

[0063] The aging mouse model was obtained by intraperitoneal injection of 200 mg / kg of D-galactose, and the blank group mice were intraperitoneally injected with an equal volume of saline. The solid-liquid mixture CTS-nL prepared as described in the previous example was taken, and the solid-liquid mixture (0.2 mL / d) was gavaged once a day at the same time as the model was established for 6 weeks. The blank group and the model group were given an equal amount of sterile water. The Morris water maze test was performed on the second day after the administration. The results are shown in Table 3, and the average value of each group was taken.

[0064] Table 3. Water maze test results Compared with the model group, the latency of crossing the platform in the CTS-1-L group was significantly shortened (P<0.05), and the number of crossing the platform was significantly increased (P<0.05). This indicates that CTS-1-L can improve the learning and memory ability of aging mice to a certain extent.

[0065] Although the embodiments of the present disclosure have been described above, it is understood that the above embodiments are exemplary and do not constitute limitations to the present disclosure. A person skilled in the art may make changes, modifications and variations to the above embodiments within the scope of the present disclosure.

Claims

1. A composition for moistening the intestines and relieving constipation, lowering blood sugar and reducing weight, and resisting aging and promoting sleep, characterized in that: The composition comprises 0.05-0.2 parts by weight of recombinant collagen, 0.05-0.2 parts by weight of γ-aminobutyric acid, 1.5-2.5 parts by weight of chia seeds, 0.5-2 parts by weight of walnut kernels, 0.5-2 parts by weight of carrots, and 0.5-2 parts by weight of cassia seeds; the amino acid sequence of the recombinant collagen is shown in SEQ ID NO:

1.

2. The composition according to claim 1, characterized in that The composition comprises 0.1 parts by weight of recombinant collagen, 0.1 parts by weight of gamma-aminobutyric acid, 2 parts by weight of chia seeds, 1 part by weight of walnut kernels, 1 part by weight of carrots, and 1 part by weight of cassia seeds.

3. The composition according to claim 1, characterized in that The chia seeds are chia seed powder, the walnut kernels are walnut kernel powder, the carrots are carrot powder, and the cassia seeds are cassia seed powder.

4. The composition according to claim 1, characterized in that The composition comprises a suitable amount of water.

5. The composition according to claim 1, characterized in that The composition is a medicine and food composition.

6. A protein that can moisten the intestines and promote bowel movements, reduce blood sugar and lose weight, and resist aging and promote sleep, characterized in that: The amino acid sequence of the protein is shown in SEQ ID NO:

1.

7. A drug for moistening the intestines and relieving constipation, lowering blood sugar and reducing weight, and resisting aging and promoting sleep, characterized in that: The drug comprises the composition according to any one of claims 1 to 5 or the protein according to claim 6.

8. A beverage that can moisten the intestines and promote bowel movements, reduce blood sugar and lose weight, and resist aging and promote sleep, characterized in that: The beverage comprises the composition according to any one of claims 1 to 5 or the protein according to claim 6.

9. A food for moistening the intestines and relieving constipation, lowering blood sugar and reducing weight, and resisting aging and promoting sleep, characterized in that: The food comprises the composition according to any one of claims 1 to 5 or the protein according to claim 6.

10. Use of the composition according to any one of claims 1 to 5 or the protein according to claim 6 in the preparation of a drug for moistening the intestines and relieving constipation, lowering blood sugar and losing weight, and resisting aging and promoting sleep.

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