Composition containing ergothioneine and having sleep-aiding, whitening, anti-aging and anti-inflammatory effects and application thereof

Abstract

The invention belongs to the technical field of health-care food, and provides a composition containing ergothioneine and having sleep-aiding, whitening, anti-aging and anti-inflammatory effects and application thereof. The ergothioneine composition disclosed by the invention is prepared from ergothioneine, collagen peptide, carnosine, nicotinamide, an olive leaf extract and a chamomile extract. The components in the composition are matched for use, the synergistic effect is obvious, the whitening, anti-aging and anti-inflammatory effects are obviously improved, meanwhile, the composition also has a sleep-aiding effect, and the comprehensive performance is excellent.

Description

Technical Field

[0001] The present invention belongs to the technical field of health food, and provides a composition containing ergothioneine and having sleep-aiding, whitening, anti-aging and anti-inflammatory effects, and application thereof. Background Art

[0002] Ergothioneine (mercaptohistidine trimethyl salt, ergothioneine, EGT) is a natural antioxidant that can protect cells in the human body and is an important active substance in the body. Ergothioneine chelates divalent iron ions and copper ions to prevent H2O2 from generating -OH under the action of iron ions or copper ions. It can also inhibit the oxidation of copper-dependent oxygenated hemoglobin and the peroxidation reaction of arachidonic acid after myoglobin (or hemoglobin) is mixed with H2O2; it can also remove hypochlorous acid, thereby preventing the inactivation of α1-antiproteinase and protecting red blood cells from the harm of neutrophils from normal function or pathological inflammatory sites; in addition, it can also inhibit the oxidation of amino acids mediated by peroxynitrite anions, such as tyrosine nitration, thereby providing feasibility for the treatment of inflammation. It has multiple physiological functions such as scavenging free radicals, detoxification, maintaining DNA biosynthesis, normal cell growth and cellular immunity. Therefore, it has a wide range of applications in the field of functional health foods.

[0003] Chinese invention patent application CN117959397A discloses a whitening and anti-aging composition containing PQQ and its application in oral whitening preparations, the components including PQQ and ergothioneine in a mass ratio of 1-6:0.5-6. PQQ and ergothioneine can play a synergistic role; when the two are used together, they can play a whitening and anti-aging effect by synergistically inhibiting tyrosinase activity and scavenging oxygen free radicals; at the same time, the composition can significantly improve the whiteness and elasticity of the skin of female subjects, increase the skin moisture content, and reduce the skin oil content, and has a significant improvement effect on the skin condition of women.

[0004] Chinese invention patent application CN117838558A discloses an anti-aging composition and a preparation method thereof. The composition includes ergothioneine, glutathione and vitamins. The composition has synergistic antioxidant, anti-aging and repair effects. Applying it to cosmetics and health products can promote skin cell metabolism, target and revitalize liver cells, enhance liver power, resist ultraviolet rays, and scavenge free radicals.

[0005] It can be seen that ergothioneine has been studied in the field of whitening and anti-aging. Chinese invention patent application CN114599358A discloses a composition for improving sleep containing ergothioneine or its salt, with L-ergothioneine or its salt as an effective ingredient, shortening the waiting time for falling asleep, reducing the time and number of waking up midway, and increasing the ratio of non-rapid eye movement sleep time to overall sleep time, etc.

[0006] Lack of sleep can also affect the body's metabolic functions, leading to dull skin and aging. At present, there are relatively few studies on ergothioneine in the field of improving sleep. At present, whitening, anti-aging and sleep aid are both hot topics in the field of functional health foods. Therefore, through the selection of functional raw materials and reasonable combination with ergothioneine, the development of a composition with both whitening, anti-aging and sleep improvement effects will be more valuable. Summary of the invention

[0007] The present invention aims at the problems existing in the prior art, and provides a composition containing ergothioneine with sleep-aiding, whitening, anti-aging and anti-inflammatory effects and its application. The composition has simple ingredients, is green and safe, and each component cooperates with each other, has the effects of whitening, anti-aging, anti-inflammatory and sleep-aiding, has excellent comprehensive performance, and is widely used.

[0008] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0009] The invention provides a composition containing ergothioneine and having the effects of aiding sleep, whitening, anti-aging and anti-inflammatory. The composition comprises: ergothioneine, collagen peptide, carnosine, nicotinamide, olive leaf extract and chamomile extract.

[0010] Furthermore, the composition comprises, by weight, 1-30 parts of ergothioneine, 200-500 parts of collagen peptides, 40-80 parts of carnosine, 0.1-15 parts of niacinamide, 80-200 parts of olive leaf extract and 100-200 parts of chamomile extract.

[0011] Furthermore, the composition comprises, by weight, 10-20 parts of ergothioneine, 200-400 parts of collagen peptides, 50-80 parts of carnosine, 1-10 parts of niacinamide, 100-150 parts of olive leaf extract and 120-150 parts of chamomile extract.

[0012] Furthermore, the composition comprises, by weight, 15-18 parts of ergothioneine, 240-280 parts of collagen peptides, 50-60 parts of carnosine, 2-5 parts of niacinamide, 120-140 parts of olive leaf extract and 130-150 parts of chamomile extract.

[0013] As a more preferred technical solution of the present invention, the ergothioneine composition further comprises: PQQ and wolfberry extract.

[0014] Furthermore, the weight ratio of ergothioneine to PQQ is 1:0.05-1.

[0015] Furthermore, the weight ratio of ergothioneine, chamomile extract and wolfberry extract is 1:10-20:5-10.

[0016] The composition of the present invention uses thioneine as the main active ingredient, because it is widely present in the human body, is an essential component of the human body but cannot be synthesized by itself, and will not produce dependence; it can carry OCTTN1 through the cell membrane to enter the cell mitochondria to clear the antioxidant of free radicals, and the only action pathway is clear, so that it can improve sleep, replenish qi and nourish the skin, and delay aging; currently there are related studies showing that thioneine can prevent sleep disorders caused by pressure, and thioneine is used in combination with collagen peptides and carnosine with the effect of scavenging active free radicals, so that its antioxidant performance is significantly improved. Related studies have shown that olive leaf extract has a certain effect in the treatment of skin diseases and trauma, so it is used in conjunction with thioneine to play a synergistic role in whitening and anti-oxidation. Secondly, with niacinamide, it promotes the synthesis of proteins such as keratin and collagen, and accelerates their differentiation, thus playing an anti-aging role; chamomile contains apigenin, an antioxidant that can bind to certain receptors in the brain, promote sleepiness and reduce insomnia or chronic insomnia; at the same time, chamomile extract can soothe the skin well, resist skin sensitivity, and has anti-inflammatory, antibacterial and antioxidant effects. The synergistic combination of each component can give full play to its own efficacy.

[0017] In addition, wolfberry is rich in various nutrients, including vitamin C, vitamin B group, zinc, iron, magnesium and various antioxidants, etc., to maintain physical health and normal function of the nervous system; in recent years, PQQ has become one of the widely-watched "star" nutrients, and its biological functions are mainly concentrated in two aspects. First, it can support the growth and development of mitochondria and stimulate the rapid growth of human cells; secondly, it has good antioxidant properties, which can help remove free radicals and reduce cell damage. As the preferred technical solution of the present invention, the addition of wolfberry extract and PQQ, in conjunction with ergothioneine, can further promote sleep quality and improve the antioxidant properties of the composition.

[0018] The present invention also provides the use of any of the above-mentioned compositions in the preparation of food or health products with sleep-aiding, whitening, anti-aging and anti-inflammatory effects.

[0019] Furthermore, the food or health product comprises any one of the above-mentioned ergothioneine compositions and a food-acceptable auxiliary material.

[0020] Furthermore, the food or health product includes any one of dairy products, instant products, beverages and tea bag preparations.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The invention has good anti-aging effect by using ergothioneine in combination with collagen peptide and carnosine, and under the synergistic effect of ergothioneine and olive leaf extract; the presence of olive leaf extract and chamomile extract gives it excellent anti-inflammatory performance; nicotinamide and carnosine have a significant synergistic effect on improving the whitening effect of the composition, and the product has a sleep-aiding effect by using ergothioneine and chamomile extract in combination; the composition, under the cooperation of various components, makes full use of the antioxidant and sleep-improving effects of ergothioneine, and makes up for the problem that the prior art composition can only have a single whitening or sleep-aiding effect and its activity is greatly limited; in addition, the invention further compounded PQQ and wolfberry extract as a preferred scheme, and the corresponding effects are further improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a picture of zebrafish neutrophil staining in Example 1 (a, model control group; b, positive control group; c, composition in Example 1);

[0024] Figure 2 This is the statistical result of zebrafish neutrophil data;

[0025] Figure 3 The relative expression results of zebrafish elastin gene in Example 1 (* compared with the blank control group, p < 0.05);

[0026] Figure 4 The relative expression results of type I collagen gene in zebrafish in Example 1 (* compared with the blank control group, p<0.05). DETAILED DESCRIPTION

[0027] The embodiments of the present invention will be described in detail below in conjunction with examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. In the examples, if no specific experimental steps or conditions are indicated, the operation or conditions of the conventional experimental steps described in the literature in this area can be carried out. If the manufacturer is not indicated for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially, and the sources of the raw materials used are shown in Table 1.

[0028] Table 1

[0029] raw material factory Specification Ergothioneine Jiangsu Jinsan Biotechnology Co., Ltd. Purity 99.99%

[0030] Examples 1-4

[0031] The raw materials were directly mixed according to the formulation in Table 2 to obtain compositions A1-A5.

[0032] Table 2 Composition formula table (unit: mg)

[0033]

[0034]

[0035] Comparative Examples 1-6

[0036] The raw materials were directly mixed according to the formula in Table 3 to obtain compositions B1-B9.

[0037] Table 3 Composition formula table (unit: mg)

[0038]

[0039] Test Example 1: Verification of anti-inflammatory, whitening and anti-aging effects

[0040] 1. Experimental materials:

[0041] (1) Experimental animals: Wild-type AB strain zebrafish (Danio rerio) aged 6 months from the National Zebrafish Resource Center of China were used for spawning tests. They were brought into the laboratory and domesticated (water temperature 26-28.5°C, room temperature 20-25°C, 1-2 fish per liter of water, and fixed light intensity of 12-16 h per day, and fed at least twice a day, including at least 1 brine shrimp larvae, with a feeding interval of more than 3 hours) for more than 14 days. Male and female fish were placed in a spawning tank at a ratio of 2:1 1-2 h before turning off the lights on the day before spawning. Healthy zebrafish embryos were selected and cultured at 28±1°C at a density of no more than 1 embryo in 200 μL of fish embryo culture medium until 6 days after fertilization.

[0042] (2) Test substance: Take compositions A1-A5 and compositions B1-B9, add fish embryo culture medium to make up to 5 mL to prepare test substance solutions.

[0043] 2. Experimental methods:

[0044] (1) Zebrafish embryo neutrophil aggregation inhibition experiment

[0045] With reference to T / HPCIA 003-2023 "Cosmetic Soothing Effect-Zebrafish Embryo Neutrophil Test Method", copper sulfate was used to induce damage to the neuromast cells in the lateral line area of ​​zebrafish embryos to cause neutrophil aggregation. Six-day-old zebrafish were divided into 17 groups, with 24 fish in each group, and exposed to 5mL fish embryo culture medium (blank control group), 10μM copper sulfate fish embryo culture medium (model control group), 10μM copper sulfate working solution + 10μM indomethacin fish embryo culture medium (positive control group), and 10μM anhydrous copper sulfate test solution (experimental groups 1-14). After 40 minutes of exposure, the fish embryos were fixed and stained with Sudan black, and the number of neutrophils in the lateral line area was counted and statistically analyzed.

[0046] (2) Experiment on promoting expression of zebrafish elastin gene (Elna)

[0047] Six-day-old zebrafish were divided into 17 groups, 36 in each group, and evenly distributed into 24-well plates. Three wells were set, each containing 12 zebrafish. The plates were exposed to 5 mL of fish embryo culture medium (blank control group), 10 μM copper sulfate fish embryo culture medium (model control group), 10 μM acetyl hexapeptide-8 working solution (positive control group), and test solution (experimental groups 1-14), respectively. After 24 hours of exposure, RNA was extracted from zebrafish, cDNA was synthesized, and real-time PCR amplification was performed. β-actin was used as the housekeeping gene, Ct was used as the amplification result, and the relative expression of Elna was calculated and statistically analyzed.

[0048] (3) Experiment on promoting the expression of type I collagen gene in zebrafish

[0049] Six-day-old zebrafish were divided into 17 groups, 36 in each group, and evenly distributed into 24-well plates. Three wells were set, each containing 12 zebrafish. The zebrafish were exposed to 5 mL of fish embryo culture medium (blank control group), 10 μM copper sulfate fish embryo culture medium (model control group), 10 μM acetyl hexapeptide-8 working solution (positive control group), and test solution (experimental groups 1-14), respectively. After 24 h of exposure, RNA was extracted from the zebrafish, cDNA was synthesized, and real-time PCR amplification was performed. β-actin was used as the housekeeping gene and Ct was used as the amplification result. The relative expression levels of col1a1a, col1a1b and col1a2 were calculated and statistically analyzed.

[0050] (4) Tyrosinase inhibition test

[0051] In the biosynthesis of melanin in the skin, tyrosinase is a key enzyme that acts on dopa to form dopaquinone, which spontaneously undergoes a series of reactions to finally form melanin. Therefore, inhibiting tyrosinase activity can reflect the whitening effect of the test sample.

[0052] Test method: The tyrosinase inhibition activity of the prepared test samples was tested according to T / SHRH 015 "Cosmetics - Tyrosinase Activity Inhibition Test Method".

[0053] 3. Experimental results:

[0054] The above compositions inhibited the aggregation of zebrafish embryonic neutrophils and promoted the expression of elastin gene (Elna) and type I collagen gene in Tables 5-8, respectively. Taking the composition of Example 1 as an example, the zebrafish neutrophil staining diagram and neutrophil data statistics are shown in Tables 5-8. Figure 1 and Figure 2 The relative expression results of zebrafish elastin gene and zebrafish type I collagen gene are shown in Figure 3 and Figure 4 .

[0055] Table 5

[0056]

[0057] Table 6

[0058]

[0059]

[0060] Table 7

[0061]

[0062] Table 8

[0063]

[0064]

[0065] According to the data in the above tables, the compositions of Examples 1-3 have excellent anti-inflammatory properties. After adding PQQ and wolfberry extract to Examples 4 and 5, the neutrophil aggregation inhibition rate is as high as 99%, and the promotion rates of elastin and collagen gene expression are further improved. Since PQQ has good antioxidant properties, this result should be directly related to the addition of PQQ, and can be used as a further preferred embodiment of the present invention.

[0066] As shown in Table 5, in terms of anti-inflammatory, by comparing Comparative Examples 4 and 5 with Example 5, it can be seen that chamomile extract and olive leaf extract have significant synergistic effects; by comparing Comparative Examples 6 and 7 with Example 5, it can be seen that ergothioneine and PQQ also have certain synergistic effects. In terms of anti-aging, by comparing Comparative Examples 1-3 with Example 5, it can be seen that ergothioneine, collagen peptides and carnosine have significant synergistic effects.

[0067] Test Example 2: Sleep-aiding efficacy verification

[0068] 1. Experimental materials:

[0069] (1) Experimental animals: SPF grade Balb / C mice, healthy adults, half male and half female, weighing 20±2 g. Purchased from Changchun Yisi Experimental Animal Technology Co., Ltd., animal production certificate number: SCXK(Ji)-2020-0002. After 7 days of adaptive feeding, the experimental period began.

[0070] (2) Experimental drugs: The experimental group used compositions A1-A5 and compositions B1-B9; positive drug: Kangxin oral liquid extract (extract during the production process of Kangxin oral liquid, which is produced by Jilin Huakang Pharmaceutical Co., Ltd., batch number 210101); blank control: normal saline.

[0071] 2. Experimental methods:

[0072] (1) Experimental grouping: After the adaptation period, 170 mice in good health were selected and divided into 17 groups according to the random number table method, with 10 mice in each group, namely blank control group, model control group, positive control group, and experimental groups 1-14.

[0073] (2) Animal model construction and drug administration:

[0074] Preventive administration: The drugs were administered at around 9 am every day. The drug-treated group was given by gavage, and the blank control group and the model group were given by gavage with an equal volume of normal saline. The administration volume was 10 mL / kg for 7 consecutive days.

[0075] Insomnia model construction: Except for the blank control group, which was intraperitoneally injected with an equal volume of saline, all other groups were intraperitoneally injected with pCPA (p-chlorophenylacetic acid) saline solution (350 mg / kg) once a day for 4 days to establish an insomnia model. Compared with the blank control group, mice showed restlessness, increased excitability, increased aggression, continuous activity, loss of circadian rhythm, decreased appetite, increased urine and feces production, gray and messy hair, and weight loss, which preliminarily indicated the success of the model. Samples of each test group were prepared at 200 mg / kg with 0.5% CMC-Na, stored at 4°C, and mixed before use every day. Samples of each group were stored at 4°C and mixed before use every day.

[0076] Therapeutic administration: The positive control group was administered with 20 g of Kangxin oral liquid extract, and the experimental groups 1-14 were administered with 5 g / kg (compositions of Examples 1-5 and Comparative Examples 1-9) by intragastric administration for 7 consecutive days, once a day. The model group and the blank group were administered with the same volume of normal saline.

[0077] (3) Testing indicators:

[0078] 3.1 Effect on the body weight of mice: Observe the behavioral changes of mice daily, weigh and record the weight of mice, and compare the weight changes of mice at different drug administration stages.

[0079] 3.2 Determination of the effect of subthreshold dose on the sleeping rate of mice: fasting but not drinking water for 8 hours before the last administration. 30 minutes after the last administration, intraperitoneal injection of subthreshold dose of sodium pentobarbital (maximum dose that can maintain the righting reflex of 90-100% mice) was given to mice in each group. Mice whose righting reflex disappeared for more than 1 minute within 30 minutes were considered to have fallen asleep, and the number of sleeping mice was recorded.

[0080] 3.3 Determination of the effect of suprathreshold dose of sodium pentobarbital on the sleep latency and sleep duration of mice: Fasting but not water for 8 hours before the last administration, 30 minutes after the last administration, each group of mice was intraperitoneally injected with a hypnotic dose (the minimum dose that makes 100% of the mice fall asleep). The time from intraperitoneal injection of sodium pentobarbital to the disappearance of the righting reflex of the mice was taken as the sleep latency, and the time from the disappearance to the recovery of the righting reflex was taken as the sleep time of the mice. The sleep latency and sleep duration of the mice were observed and recorded.

[0081] (4) Experimental results:

[0082] The results of the measurements of mouse body weight, the effect of subthreshold dose on the sleep rate of mice, and the effect of suprathreshold dose of sodium pentobarbital on the sleep latency and sleep time of mice are recorded in Tables 9-12, respectively.

[0083] Table 9

[0084]

[0085]

[0086] Table 10

[0087]

[0088]

[0089] Table 11

[0090] Sleep latency (min) Blank control group 2.11±1.25 Model control group <![CDATA[12.43±2.45 ## ]]> Positive control group <![CDATA[7.42±1.58 # *]]> Example 1 <![CDATA[3.21±1.06** ^ ]]> Example 2 <![CDATA[2.98±1.41** ^ ]]> Example 3 <![CDATA[3.46±1.33** ^ ]]> Example 4 <![CDATA[3.15±1.12** ^ ]]> Example 5 <![CDATA[3.08±1.04** ^ ]]> Comparative Example 1 9.32±1.47**^ Comparative Example 2 <![CDATA[3.95±1.57** ^ ]]> Comparative Example 3 <![CDATA[3.65±1.44** ^ ]]> Comparative Example 4 <![CDATA[7.05±1.83 # * ％& ]]> Comparative Example 5 <![CDATA[3.67±1.55 # ** ^ ]]> Comparative Example 6 <![CDATA[8.05±1.62 # * ％& <!-- 10 -->]]> Comparative Example 7 <![CDATA[4.06±1.70 # **^]]> Comparative Example 8 <![CDATA[3.86±1.07** ^ ]]> Comparative Example 9 <![CDATA[5.54±1.25** ^％& ]]>

[0091] Note: #Compared with blank group: P<0.05, ##Compared with blank group: P<0.01; *Compared with model group: P<0.05, **Compared with model group: P<0.01; ^Compared with positive control group: P<0.05, ^^Compared with positive control group: P<0.01; %Compared with experimental group 1: P<0.05, %%Compared with experimental group 1: P<0.01; &Compared with experimental group 5: P<0.05, &&Compared with experimental group 5: P<0.01.

[0092] Table 12

[0093]

[0094]

[0095] Note: #Compared with blank group: P<0.05, ##Compared with blank group: P<0.01; *Compared with model group: P<0.05, **Compared with model group: P<0.01; ^Compared with positive control group: P<0.05, ^^Compared with positive control group: P<0.01; %Compared with experimental group 1: P<0.05, %%Compared with experimental group 1: P<0.01; &Compared with experimental group 5: P<0.05, &&Compared with experimental group 5: P<0.01.

[0096] According to the above data in each table, the model control group is compared with the blank control group, and the sleep latency is extended, and the sleep time decreases, and there is a very significant difference, indicating that the modeling is successful. In the present invention, the medicine of embodiment 1-5, comparative example 1-9 and positive control group can make mice weight growth compared to blank control group and model control group, but there is no significant difference. Wherein, compared with the embodiment, comparative example 1,6 mice sleep rate is the lowest, the sleep latency is the longest, and the sleep time is the shortest, and the comparative example 4 is the second, indicating that in the present composition, ergothioneine and chamomile extract play a key role in improving sleep. Each embodiment is compared with the positive control drug, and the mouse sleep rate decreases, and the sleep latency significantly decreases, and the sleep time is significantly improved, indicating that the composition of the present invention has the effect of significantly improving sleep, and the effect is better than positive drug Kangxin oral liquid.

[0097] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A composition containing ergothioneine having sleep-aiding, whitening, anti-aging and anti-inflammatory effects, characterized in that: The composition comprises: ergothioneine, collagen peptide, carnosine, niacinamide, olive leaf extract and chamomile extract.

2. The composition according to claim 1, characterized in that: The composition comprises, by weight, 1-30 parts of ergothioneine, 200-500 parts of collagen peptide, 40-80 parts of carnosine, 0.1-15 parts of nicotinamide, 80-200 parts of olive leaf extract and 100-200 parts of chamomile extract.

3. The composition according to claim 2, characterized in that: The composition comprises, by weight, 10-20 parts of ergothioneine, 200-400 parts of collagen peptide, 50-80 parts of carnosine, 1-10 parts of nicotinamide, 100-150 parts of olive leaf extract and 120-150 parts of chamomile extract.

4. The composition according to claim 3, characterized in that: The composition comprises, by weight, 15-18 parts of ergothioneine, 240-280 parts of collagen peptide, 50-60 parts of carnosine, 2-5 parts of nicotinamide, 120-140 parts of olive leaf extract and 130-150 parts of chamomile extract.

5. The composition according to any one of claims 1 to 4, characterized in that: Also includes: PQQ and wolfberry extract.

6. The composition according to claim 5, characterized in that: The weight ratio of ergothioneine to PQQ is 1:0.05-1.

7. The composition according to claim 5, characterized in that: The weight ratio of the ergothioneine, chamomile extract and wolfberry extract is 1:10-20:5-10.

8. Use of the composition according to any one of claims 1 to 7 in the preparation of food or health products with sleep-aiding, whitening, anti-aging and anti-inflammatory effects.

9. The use according to claim 8, characterized in that: The food or health product comprises the ergothioneine composition described in any one of claims 1 to 7 and a food-acceptable auxiliary material.

10. The use according to claim 8, characterized in that: The food or health product includes any one of dairy products, instant products, beverages and tea bag preparations.

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