Composition with anti-inflammatory and anti-aging effects and application thereof

Abstract

The invention relates to a composition with anti-inflammatory and anti-aging effects and application thereof, the composition comprises a dogwood extract, a radix fici simplicissimae extract and a guava extract, the composition can effectively scavenge free radicals, inhibit collagenase activity and inhibit generation of fluorescent AGEs, and the composition has the anti-inflammatory and anti-aging effects. Skin aging and skin laxity and darkness caused by glycosylation reaction are prevented. Meanwhile, by inhibiting NO secretion, the composition relieves skin inflammation reaction and can also improve skin microcirculation. All the components have a synergistic effect, so that the anti-oxidation, anti-inflammatory, anti-aging and anti-saccharification effects are remarkably enhanced, and the skin problems are comprehensively solved. No organic solvent is used in the process, the green and environment-friendly concept is met, the safety and reliability of the extract are ensured, and a mild and efficient solution is provided for anti-aging.

Description

Technical Field

[0001] The invention belongs to the technical field of plant extraction, and relates to a composition with anti-inflammatory and anti-aging effects and application thereof. Background Art

[0002] Skin aging is a complex and gradual physiological process, which is affected by the interaction of multiple internal and external factors. External factors such as human metabolism and ultraviolet radiation can lead to the massive generation of free radicals. These free radicals are highly oxidative and can attack various biological molecules in skin cells, affecting the renewal and repair of skin cells, and ultimately accelerating the aging process of the skin. When the body's balance is disturbed, the activity of collagenase is disturbed, and collagen is over-decomposed, resulting in a decrease in the content of collagen in the skin, loosening the skin fiber structure and losing elasticity. At the same time, the accumulation of AGEs (advanced glycation end products) in the skin not only makes the skin stiff and loses elasticity, but also aggravates the dullness and yellowing of the skin. Excessive AGEs induce oxidative stress reactions, further increase the generation of free radicals, stimulate the release of inflammatory factors, activate the expression of collagenase, form a vicious cycle, and accelerate skin aging and damage.

[0003] Based on this, the present invention aims to provide a composition with anti-inflammatory and anti-aging effects and its application, aiming to improve the aging state of the skin through multi-faceted regulation, inhibit the generation of free radicals and the accumulation of AGEs, thereby effectively preventing and alleviating skin aging-related problems. Summary of the invention

[0004] In view of the deficiencies of the prior art, the purpose of the present invention is to provide a composition with anti-inflammatory and anti-aging effects and its application. By scientifically selecting natural plant extracts with significant antioxidant, anti-aging and anti-inflammatory effects and combining their synergistic effects, the present invention can improve skin health in multiple dimensions and all directions, and prevent and delay the skin aging process.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a composition with anti-inflammatory and anti-aging effects, comprising a Cornus officinalis extract, a Prunus armeniaca extract and a Psidium guajava extract.

[0007] Cornus officinalis Sieb.et Zucc. is a plant of the genus Cornus in the family Cornaceae. Studies have found that the pulp of Cornus officinalis is rich in iridoid glycosides, triterpenes, and polysaccharides, which have beneficial effects on the human body, such as lowering blood sugar, improving immunity, anti-tumor, anti-inflammatory, antibacterial, and antioxidant.

[0008] Ficus hirta Vahl. is a plant of the Moraceae family and a traditional medicinal and edible plant in South China. Its main chemical components include coumarins, polysaccharides, steroids, phenols and flavonoids, which have multiple pharmacological effects such as anti-inflammatory, analgesic, immunity-enhancing, liver-protecting, antibacterial and antioxidant.

[0009] Guava (Psidium guajava L.) is a tree plant of the genus Guava in the Myrtaceae family. It is rich in various nutrients, including polyphenols, flavonoids, polysaccharides, vitamins, trace elements, etc. These substances play an important role in physical health and beauty. In addition, guava can effectively promote the balance of blood sugar in the human body and have a positive impact on the prevention and management of chronic diseases.

[0010] Cornus officinalis, ginseng, and guava are all raw materials that can be used as medicine and food. These natural ingredients can comprehensively improve skin health through scientific proportions and synergistic effects. They can not only improve skin condition through direct antioxidant and anti-inflammatory effects, but also fundamentally prevent and delay the skin aging process by regulating the body's internal physiological functions.

[0011] Preferably, the composition comprises, by mass, 10-50 parts of Cornus officinalis extract, 20-60 parts of Eleutherodactyla quinata extract and 10-50 parts of Psidium guajava extract.

[0012] The mass fraction of the Cornus officinalis extract can be selected from 10 parts, 25 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts, etc., the mass fraction of the Elaeagnus quinquefolius extract can be selected from 20 parts, 25 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts, 55 parts, 60 parts, etc., and the mass fraction of the guava extract can be selected from 10 parts, 15 parts, 20 parts, 25 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts, etc. Other specific point values ​​within the above numerical range can be selected, and they will not be listed here one by one.

[0013] Preferably, the Cornus officinalis extract is prepared by a method comprising the following steps:

[0014] (1) Mixing Cornus officinalis with water and performing ultra-high pressure microfluidization pretreatment;

[0015] (2) mixing the pretreated Cornus officinalis with an enzyme for enzymolysis to obtain an enzymolysis solution;

[0016] (3) The enzymatic hydrolyzate is filtered, concentrated, and dried to obtain the product.

[0017] Enzymatic hydrolysis can assist in releasing the active ingredients in the cells. By controlling the enzymatic hydrolysis conditions, the enzymatic hydrolysis efficiency can be maximized to avoid destroying the active ingredients.

[0018] Preferably, the Prunus armeniaca extract is prepared by a method comprising the following steps:

[0019] (1) mixing the five-fingered peach with water, and extracting under ultrasound to obtain an extract;

[0020] (2) Filter the extract, concentrate it, and dry it to obtain the product.

[0021] Ultrasonic waves can generate strong vibration and cavitation effects, which can help destroy cell structures and accelerate the release and extraction of active ingredients. By controlling ultrasonic conditions, the extraction efficiency and product purity can be maximized.

[0022] Preferably, the guava extract is prepared by a method comprising the following steps:

[0023] (1) mixing guava with water and performing ultra-high pressure microfluidization pretreatment;

[0024] (2) subjecting the pretreated guava to reflux extraction to obtain an extract;

[0025] (3) The extract is centrifuged to obtain the supernatant, which is filtered, concentrated, and dried to obtain the product.

[0026] Preferably, the liquid ratio of Cornus officinalis to water is 1g:(10-30)mL, wherein 10, 15, 20, 25, 30, etc. in (10-30) and other specific point values ​​within the above numerical range can be selected, which will not be repeated here.

[0027] Preferably, the ultra-high pressure microfluidic pretreatment is performed at a pressure of 100-300 MPa, a temperature of 60-80° C., and a number of times of 2-6 times.

[0028] The pressure can be selected from 100MPa, 120MPa, 140MPa, 160MPa, 180MPa, 200MPa, 220MPa, 240MPa, 260MPa, 280MPa, 300MPa, etc., the temperature is 60°C, 62°C, 64°C, 66°C, 68°C, 70°C, 72°C, 74°C, 76°C, 78°C, 80°C, etc., the number of times can be selected from 2 times, 3 times, 4 times, 5 times, 6 times, etc., other specific point values ​​within the above numerical range can be selected, and they will not be elaborated here.

[0029] Preferably, the enzyme is a combination of cellulase and pectinase, and the amount of the enzyme added is 2-5% of the mass of Cornus officinalis, for example 2%, 2.2%, 2.5%, 2.8%, 3%, 3.2%, 3.5%, 3.8%, 4%, 4.2%, 4.5%, 4.8%, 5%, etc. Other specific point values ​​within the above numerical range can be selected, which will not be repeated here.

[0030] Preferably, the enzyme activity ratio of cellulase and pectinase is 1:(1-2), for example 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, etc. Other specific points within the above numerical range can be selected and will not be described here one by one.

[0031] Preferably, the enzymatic hydrolysis temperature is 35-55°C, the pH is 3-5.5, and the time is 20-30 min.

[0032] The temperature can be selected from 35°C, 37°C, 40°C, 42°C, 44°C, 46°C, 48°C, 50°C, 52°C, 55°C, etc., the pH can be selected from 3, 3.2, 3.5, 3.8, 4, 4.2, 4.5, 4.8, 5, 5.2, 5.5, etc., the time can be selected from 20min, 21min, 22min, 23min, 24min, 25min, 26min, 27min, 28min, 29min, 30min, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be described one by one here.

[0033] Preferably, the enzymatic hydrolysis further comprises keeping warm at 78-85°C for 30-50 minutes.

[0034] The temperature can be selected from 78°C, 79°C, 80°C, 81°C, 82°C, 83°C, 84°C, 85°C, etc., and the time can be selected from 30min, 32min, 34min, 36min, 38min, 40min, 42min, 44min, 46min, 48min, 50min, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0035] Preferably, the filtration comprises filtering through a sieve, a ceramic membrane and an ultrafiltration membrane in sequence.

[0036] Preferably, the aperture of the sieve is 400-600 mesh, for example, 400 mesh, 420 mesh, 440 mesh, 460 mesh, 480 mesh, 500 mesh, 520 mesh, 540 mesh, 560 mesh, 580 mesh, 600 mesh, etc. Other specific point values ​​within the above numerical range can be selected, and they will not be described one by one here.

[0037] Preferably, the pore size of the ceramic membrane is 100-200nm, for example, 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, 160nm, 170nm, 180nm, 190nm, 200nm, etc. Other specific point values ​​within the above numerical range can be selected and will not be described here one by one.

[0038] Preferably, the pore size of the ultrafiltration membrane is 1000-5000Da, for example 1000Da, 1500Da, 2000Da, 2500Da, 3000Da, 3500Da, 4000Da, 4500Da, 5000Da, etc. Other specific point values ​​within the above numerical range can be selected, which will not be described here one by one.

[0039] Preferably, the vacuum degree of the concentration is 60-110 mbar and the temperature is 50-60°C.

[0040] The vacuum degree can be selected as 60mbar, 65mbar, 70mbar, 75mbar, 80mbar, 85mbar, 90mbar, 95mbar, 100mbar, 105mbar, 110mbar, etc., and the temperature can be selected as 50℃, 51℃, 52℃, 53℃, 54℃, 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0041] Preferably, the liquid ratio of five-fingered peach to water is 1g:(10-30)mL, where 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, etc. in (10-30) and other specific point values ​​within the above numerical range can be selected, which will not be repeated here.

[0042] Preferably, before the extraction, the method further comprises treating the mixture of the quinquefolius and water at 110-130° C. and 0.1-0.2 MPa for 15-30 min.

[0043] The temperature can be selected as 110℃, 112℃, 114℃, 116℃, 118℃, 120℃, 122℃, 124℃, 126℃, 128℃, 130℃, etc. The pressure can be selected as 0.1MPa, 0.11MPa, 0.12MPa, 0.13MPa, 0.14MPa, 0.15MPa0.16MPa, 0.17MPa, 0.18MPa, 0.19MPa, 0.2MPa, etc. The time can be selected as 15min, 16min, 17min, 18min, 19min, 20min, 21min, 22min, 23min, 24min, 25min, 26min, 27min, 28min, 29min, 30min, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be described one by one here.

[0044] Preferably, the temperature of the ultrasonic extraction is 60-80°C, the frequency is 30-40kHz, and the time is 15-30min.

[0045] The temperature can be selected as 60℃, 62℃, 64℃, 66℃, 68℃, 70℃, 72℃, 74℃, 76℃, 78℃, 80℃, etc. The frequency can be selected as 30kHz, 31kHz, 32kHz, 33kHz, 34kHz, 35kHz, 36kHz, 37kHz, 38kHz, 39kHz, 40kHz, etc. The time can be selected as 15min, 16min, 17min, 18min, 19min, 20min, 21min, 22min, 23min, 24min, 25min, 26min, 27min, 28min, 29min, 30min, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0046] Preferably, the number of ultrasounds is 1-3 times, for example, 1 time, 2 times, 3 times, etc.

[0047] Preferably, the vacuum degree of the concentration is 60-110 mbar and the temperature is 50-60°C.

[0048] The vacuum degree can be selected as 60mbar, 65mbar, 70mbar, 75mbar, 80mbar, 85mbar, 90mbar, 95mbar, 100mbar, 105mbar, 110mbar, etc., and the temperature can be selected as 50℃, 51℃, 52℃, 53℃, 54℃, 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0049] Preferably, the liquid-to-solid ratio of guava to water is 1:(10-30) mL, wherein the specific point values ​​in (10-30) can be selected from 10, 15, 20, 25, 30, etc., and other specific point values ​​within the above numerical range can be selected, which will not be described one by one here.

[0050] Preferably, the ultra-high pressure microfluidic pretreatment is performed at a pressure of 100-300 MPa, a temperature of 60-80° C., and a number of times of 3-6 times.

[0051] The pressure can be selected from 100MPa, 120MPa, 140MPa, 160MPa, 180MPa, 200MPa, 220MPa, 240MPa, 260MPa, 280MPa, 300MPa, etc., the temperature can be selected from 60°C, 62°C, 64°C, 66°C, 68°C, 70°C, 72°C, 74°C, 76°C, 78°C, 80°C, etc., the number of times can be selected from 3 times, 4 times, 5 times, 6 times, etc., and other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0052] Preferably, the reflux extraction time is 20-40 min.

[0053] The time can be selected as 20min, 25min, 30min, 35min, 40min, etc. Other specific point values ​​within the above numerical range can also be selected, and they will not be described one by one here.

[0054] Preferably, the centrifugal speed is 5000-7000 rpm and the time is 5-10 min.

[0055] The rotation speed can be selected as 5000rpm, 5500rpm, 6000rpm, 6500rpm, 7000rpm, etc., and the time can be selected as 5min, 6min, 7min, 8min, 9min, 10min, etc. Other specific point values ​​within the above numerical range can be selected, and they will not be described here one by one.

[0056] Preferably, the filtering comprises filtering through a 100-200 nm ceramic membrane.

[0057] The ceramic membrane can be selected at 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, 160nm, 170nm, 180nm, 190nm, 200nm, etc. Other specific point values ​​within the above numerical range can be selected, which will not be described here one by one.

[0058] Preferably, the vacuum degree of the concentration is 60-110 mbar and the temperature is 50-60°C.

[0059] The vacuum degree can be selected as 60mbar, 65mbar, 70mbar, 75mbar, 80mbar, 85mbar, 90mbar, 95mbar, 100mbar, 105mbar, 110mbar, etc., and the temperature can be selected as 50℃, 51℃, 52℃, 53℃, 54℃, 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0060] Compared with the prior art, the present invention has the following beneficial effects:

[0061] (1) The present invention effectively extracts the active ingredients from Cornus officinalis, Eleutherodactyla quinquefolia and Psidium guajava through ultra-high pressure microfluidic extraction, complex enzyme hydrolysis, high pressure extraction, ultrasound-assisted extraction and the like, and successfully obtains a composition with anti-inflammatory, anti-aging, anti-glycation and microcirculation improving effects.

[0062] (2) The composition obtained by the preparation method of the present invention can significantly scavenge DPPH free radicals and inhibit collagenase activity.

[0063] (3) The composition obtained by the preparation method of the present invention can significantly inhibit the generation of fluorescent AGEs and reduce the relative expression level of NO.

[0064] (4) The composition obtained by the preparation method of the present invention can significantly promote blood circulation and improve microcirculation of zebrafish.

[0065] (5) The composition obtained by the preparation method of the present invention has synergistic effects in anti-oxidation, anti-aging, anti-inflammatory, anti-glycation and improving microcirculation.

[0066] (6) The extraction solvent of the present invention uses RO water, which can not only effectively extract the bioactive components in plants, but also further expand its application scope in the field of health. No organic solvent is used in the whole process, and no complicated recovery and treatment links are required, which greatly reduces energy consumption and pollutant emissions, effectively reduces environmental pressure, and better meets the needs of industrial production. DETAILED DESCRIPTION

[0067] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0068] The sources of the functional ingredients contained in the products involved in the following embodiments and comparative examples are as follows (only the functional ingredients are reflected, and the necessary auxiliary ingredients contained in other commercially available raw materials are not repeated):

[0069] Cellulase was purchased from Wuxi Xuemei Enzyme Preparation Technology Co., Ltd. under the trade name of Cellulase.

[0070] Pectinase was purchased from Tianjin Lihua Enzyme Preparation Technology Co., Ltd. under the trade name of Pectinase.

[0071] Preparation Example 1-1

[0072] This preparation example provides a method for preparing a Cornus officinalis extract, the preparation method comprising:

[0073] (1) 10 g of Cornus officinalis was crushed and mixed with water at a solid-liquid ratio of 1 g:20 mL. Ultrahigh pressure microfluidization pretreatment was performed at a pressure of 200 MPa and a temperature of 70°C for 4 times.

[0074] (2) The Cornus officinalis obtained in step (1) was mixed with 0.4 g of the complex enzyme and subjected to enzymatic hydrolysis at 45° C. and pH 4 for 25 min. After the enzymatic hydrolysis, the mixture was kept at 80° C. for 40 min.

[0075] (3) The product after enzymatic hydrolysis was filtered through a 400-mesh screen, a 5 μm filter bag, a 100 nm ceramic membrane, and a 1000 Da ultrafiltration membrane in sequence, and the retentate was concentrated to 15 wt % at a temperature of 55° C. and a vacuum degree of 80 mbar, sterilized, and freeze-dried to obtain the product.

[0076] Preparation Example 1-2

[0077] This preparation example provides a method for preparing a Cornus officinalis extract, the preparation method comprising:

[0078] (1) 10 g of Cornus officinalis was crushed and mixed with water at a solid-liquid ratio of 1 g:10 mL. Ultrahigh pressure microfluidization pretreatment was performed at a pressure of 100 MPa and a temperature of 60° C. for 2 times.

[0079] (2) The Cornus officinalis obtained in step (1) was mixed with 0.2 g of the complex enzyme and subjected to enzymatic hydrolysis at 35° C. and pH 3 for 20 min. After the enzymatic hydrolysis, the mixture was kept at 80° C. for 30 min.

[0080] (3) The product after enzymatic hydrolysis was filtered through a 400-mesh screen, a 5 μm filter bag, a 200 nm ceramic membrane, and a 3000 Da ultrafiltration membrane in sequence, and the retentate was concentrated to 20 wt % at a temperature of 50° C. and a vacuum degree of 60 mbar, sterilized, and freeze-dried to obtain the product.

[0081] Preparation Example 1-3

[0082] This preparation example provides a method for preparing a Cornus officinalis extract, the preparation method comprising:

[0083] (1) 10 g of Cornus officinalis was crushed and mixed with water at a solid-liquid ratio of 1 g:20 mL. Ultrahigh pressure microfluidization pretreatment was performed at a pressure of 300 MPa and a temperature of 80° C. for 6 times.

[0084] (2) The Cornus officinalis obtained in step (1) was mixed with 0.5 g of a complex enzyme and subjected to enzymatic hydrolysis at 55° C. and pH 5.5 for 30 min. After the enzymatic hydrolysis, the mixture was kept at 80° C. for 50 min.

[0085] (3) The product after enzymatic hydrolysis was filtered through a 400 mesh screen, a 5 μm filter bag, a 150 nm ceramic membrane, and a 4000 Da ultrafiltration membrane in sequence, and the retentate was concentrated to 10 wt % at a temperature of 60° C. and a vacuum degree of 110 mbar, sterilized, and freeze-dried to obtain the product.

[0086] Preparation Example 1-4

[0087] This preparation example provides a preparation method of Cornus officinalis extract, which differs from Preparation Example 1-1 only in that step (1) is "crushing 10 g of Cornus officinalis and mixing it with water, and the solid-liquid ratio of Cornus officinalis to water is 1 g:20 mL", and other operations remain unchanged.

[0088] Preparation Example 1-5

[0089] This preparation example provides a preparation method of Cornus officinalis extract, which differs from Preparation Example 1-1 only in that step (2) is "treating the Cornus officinalis obtained in step (1) at 45°C and pH 4 for 25 minutes, and then keeping warm at 80°C for 40 minutes", and other operations remain unchanged.

[0090] Preparation Example 1-6

[0091] This preparation example provides a preparation method of Cornus officinalis extract, which differs from Preparation Example 1-1 only in that step (3) is "filtering the enzymatic hydrolyzate obtained in step (2) through a 400-mesh sieve and a 5 μm filter bag, concentrating to 15 wt% at a temperature of 55° C. and a vacuum degree of 80 mbar, sterilizing, and freeze-drying to obtain", and other operations remain unchanged.

[0092] Preparation Example 1-7

[0093] This preparation example provides a method for preparing a Cornus officinalis extract, which differs from Preparation Example 1-1 only in that step (3) is "filtering the enzymatic hydrolyzate obtained in step (2) through a 400-mesh sieve, a 5 μm filter bag, and a 100 nm ceramic membrane, taking the filtrate, concentrating it to 15 wt% at a temperature of 55° C. and a vacuum degree of 80 mbar, sterilizing it, and freeze-drying it to obtain the product", and other operations remain unchanged.

[0094] Preparation Example 1-8

[0095] This preparation example provides a method for preparing a Cornus officinalis extract, which differs from Preparation Example 1-1 only in that step (2) is "reflux extraction of the product obtained in step (1) for 25 minutes, twice", and other operations remain unchanged.

[0096] Preparation Example 2-1

[0097] This preparation example provides a preparation method of a Prunus armeniaca extract, the preparation method comprising:

[0098] (1) The five-fingered ginseng peach was crushed and mixed with water, the solid-liquid ratio of the five-fingered ginseng peach to water was 1 g:20 mL, and the mixture was treated at 0.15 MPa and 120° C. for 20 min.

[0099] (2) The product obtained in step (1) was subjected to ultrasonic extraction at 70° C. for 20 min, with an ultrasonic frequency of 35 kHz and a number of times of 2 times.

[0100] (3) The product obtained in step (2) is passed through a 400-mesh sieve and a 5 μm filter bag, concentrated to 10 wt % at 80 mbar and 55° C., sterilized, and dried to obtain the product.

[0101] Preparation Example 2-2

[0102] This preparation example provides a preparation method of a Prunus armeniaca extract, the preparation method comprising:

[0103] (1) The five-fingered ginseng peach was crushed and mixed with water, the solid-liquid ratio of the five-fingered ginseng peach to water was 1 g:10 mL, and the mixture was treated at 0.1 MPa and 110° C. for 30 min.

[0104] (2) The product obtained in step (1) was subjected to ultrasonic extraction at 80° C. for 15 min, with an ultrasonic frequency of 40 kHz and a number of times of 3 times.

[0105] (3) The product obtained in step (2) is passed through a 400-mesh sieve and a 5 μm filter bag, concentrated to 15 wt % at 60 mbar and 50° C., sterilized, and dried to obtain the product.

[0106] Preparation Example 2-3

[0107] This preparation example provides a preparation method of a Prunus armeniaca extract, the preparation method comprising:

[0108] (1) The five-fingered ginseng peach was crushed and mixed with water, the solid-liquid ratio of the five-fingered ginseng peach to water was 1 g:30 mL, and the mixture was treated at 0.2 MPa and 130° C. for 15 min.

[0109] (2) The product obtained in step (1) was subjected to ultrasonic extraction at 60° C. for 30 min, with an ultrasonic frequency of 30 kHz and a frequency of 1 time.

[0110] (3) The product obtained in step (2) is passed through a 400-mesh sieve and a 5 μm filter bag, concentrated to 5 wt % at 110 mbar and 60° C., sterilized, and dried to obtain the product.

[0111] Preparation Example 2-4

[0112] This preparation example provides a preparation method of the Eleutherodactyl extract, which differs from Preparation Example 2-1 only in that step (1) is "crushing the Eleutherodactyl extract and mixing it with water, with the solid-liquid ratio of Eleutherodactyl extract to water being 1 g:20 mL", and other operations remain unchanged.

[0113] Preparation Example 2-5

[0114] This preparation example provides a preparation method of a Prunus armeniaca extract, which differs from Preparation Example 2-1 only in that step (2) is "extracting the product obtained in step (1) at 70°C for 20 minutes, twice", and other operations remain unchanged.

[0115] Preparation Example 2-6

[0116] This preparation example provides a preparation method of a Prunus armeniaca extract, which differs from Preparation Example 2-1 only in that step (2) is to "reflux and extract the product obtained in step (1) for 20 minutes, twice", and other operations remain unchanged.

[0117] Preparation Example 2-7

[0118] This preparation example provides a preparation method of a Prunus armeniaca extract, which differs from Preparation Example 2-1 only in that step (2) is "extracting the product obtained in step (1) by negative pressure cavitation for 1 hour at a temperature of 40°C and a negative pressure intensity of 90 mbar", and other operations remain unchanged.

[0119] Preparation Example 3-1

[0120] This preparation example provides a preparation method of guava extract, the preparation method comprising:

[0121] (1) The guava was crushed and mixed with water in a ratio of 1 g:15 mL. The guava was pretreated with ultrahigh pressure microfluidization. The temperature of the ultrahigh pressure microfluidization treatment was 70° C., the pressure was 200 MPa, and the number of times was 5 times.

[0122] (2) refluxing and extracting the guava obtained in step (1) for 30 minutes;

[0123] (3) The product obtained by reflux extraction was centrifuged at 6000 rpm for 8 min, the supernatant was filtered through a 150 nm filter membrane, concentrated to 10 wt% at 55° C. and 80 mbar, sterilized, and dried to obtain the product.

[0124] Preparation Example 3-2

[0125] This preparation example provides a preparation method of guava extract, the preparation method comprising:

[0126] (1) The guava was crushed and mixed with water at a ratio of 1 g:10 mL. The guava was pretreated with ultrahigh pressure microfluidization at a temperature of 60° C. and a pressure of 100 MPa for 6 times.

[0127] (2) refluxing and extracting the guava obtained in step (1) for 40 minutes;

[0128] (3) The product obtained by reflux extraction was centrifuged at 7000 rpm for 5 min, the supernatant was filtered through a 100 nm filter membrane, concentrated to 15 wt% at 50° C. and 60 mbar, sterilized, and dried to obtain the product.

[0129] Preparation Example 3-3

[0130] This preparation example provides a preparation method of guava extract, the preparation method comprising:

[0131] (1) The guava was crushed and mixed with water in a ratio of 1 g:20 mL. The guava was pretreated with ultrahigh pressure microfluidization. The temperature of the ultrahigh pressure microfluidization treatment was 80° C., the pressure was 300 MPa, and the number of times was 3 times.

[0132] (2) extracting the guava obtained in step (1) by reflux for 20 minutes;

[0133] (3) The product obtained by reflux extraction was centrifuged at 5000 rpm for 10 min, the supernatant was filtered through a 200 nm filter membrane, concentrated to 5 wt% at 60° C. and 110 mbar, sterilized, and dried to obtain the product.

[0134] Example 1

[0135] This embodiment provides a composition with anti-inflammatory and anti-aging effects, which comprises, by mass, 50 parts of Cornus officinalis extract obtained in Preparation Example 1-1, 30 parts of Prunus armeniaca extract obtained in Preparation Example 2-1, and 10 parts of Psidium guajava extract obtained in Preparation Example 3-1.

[0136] The preparation method thereof is as follows: physically mixing the preparation raw materials to obtain the product.

[0137] Example 2

[0138] This embodiment provides a composition with anti-inflammatory and anti-aging effects, which comprises, by mass, 10 parts of Cornus officinalis extract obtained in Preparation Example 1-2, 20 parts of Prunus armeniaca extract obtained in Preparation Example 2-2, and 50 parts of Psidium guajava extract obtained in Preparation Example 3-2.

[0139] The preparation method is similar to that of Example 1.

[0140] Example 3

[0141] This embodiment provides a composition with anti-inflammatory and anti-aging effects, which comprises, by mass, 30 parts of Cornus officinalis extract obtained in Preparation Example 1-3, 50 parts of Prunus armeniaca extract obtained in Preparation Example 2-3, and 30 parts of Psidium guajava extract obtained in Preparation Example 3-3.

[0142] The preparation method is similar to that of Example 1.

[0143] Embodiment 4-8

[0144] This embodiment provides 5 compositions with anti-inflammatory and anti-aging effects, which differ from Example 1 only in that the Cornus officinalis extract obtained in Preparation Example 1-1 is replaced in equal amounts by the Cornus officinalis extracts obtained in Preparation Example 1-4, Preparation Example 1-5, Preparation Example 1-6, Preparation Example 1-7, and Preparation Example 1-8, and the other ingredients and contents remain unchanged.

[0145] The preparation method is similar to that of Example 1.

[0146] Examples 9-12

[0147] This embodiment provides 4 compositions with anti-inflammatory and anti-aging effects, which differ from Example 1 only in that the five-fingered peach extract obtained in Preparation Example 2-1 is replaced in equal amounts by the five-fingered peach extracts obtained in Preparation Example 2-4, Preparation Example 2-5, Preparation Example 2-6, and Preparation Example 2-7, and the other ingredients and contents remain unchanged.

[0148] The preparation method is similar to that of Example 1.

[0149] Comparative Example 1

[0150] This comparative example provides a composition with anti-inflammatory and anti-aging effects, which is different from Example 1 only in that it does not contain Cornus officinalis extract, and its reduced mass is proportionally distributed to the masses of Eleutherodendron chinense extract and Psidium guajava extract.

[0151] The preparation method is similar to that of Example 1.

[0152] Comparative Example 2

[0153] This comparative example provides a composition with anti-inflammatory and anti-aging effects, which is different from Example 1 only in that it does not contain the Eleutherodactylus quinquefolius extract, and its reduced mass is proportionally distributed to the mass of the Cornus officinalis extract and the Psidium guajava extract.

[0154] The preparation method is similar to that of Example 1.

[0155] Comparative Example 3

[0156] This comparative example provides a composition with anti-inflammatory and anti-aging effects, which is different from Example 1 only in that it does not contain guava extract, and its reduced mass is proportionally distributed to the masses of the quinquefolia extract and the cornus officinalis extract.

[0157] The preparation method is similar to that of Example 1.

[0158] Test Example 1

[0159] DPPH free radical scavenging experiment

[0160] Experimental methods:

[0161] The DPPH solution was dissolved in anhydrous ethanol to a concentration of 0.1 mg / mL, and a 0.5 mg / mL vitamin C standard (positive control) and a sample solution to be tested were prepared at the same time. 150 μL of the DPPH alcohol solution and 150 μL of the sample solution were mixed evenly, reacted at room temperature in the dark for 30 minutes, shaken and evenly, and the absorbance was measured at 517 nm. Each group was measured 3 times on average and the average was taken. The scavenging rate of the composite sample on DPPH free radicals was calculated according to the following formula, and the results are shown in Table 1.

[0162] Sample group: sample solution 150μL + DPPH alcohol solution 150μL

[0163] Sample blank group: DPPH solution 150μL + sample solvent 150μL

[0164] Control group: sample solution 150μL + anhydrous ethanol 150μL

[0165] Blank control group: 150 μL of anhydrous ethanol + 150 μL of sample solvent

[0166]

[0167] Table 1

[0168] Group Clearance rate (%) Example 1 98.71 Example 2 96.15 Example 3 95.61 Example 4 85.63 Example 5 89.37 Example 6 88.58 Example 7 89.08 Example 8 88.64 Example 9 87.45 Example 10 90.64 Embodiment 11 82.18 Example 12 81.64 Comparative Example 1 58.25 Comparative Example 2 80.67 Comparative Example 3 68.72 Preparation Example 1-1 81.13 Preparation Example 2-1 38.48 Preparation Example 3-1 71.85 Positive Control 98.74

[0169] It can be seen from the data results in Table 1 that the composition described in the present application has a good DPPH free radical scavenging effect. The preparation method of Cornus officinalis extract and Eleutherodactyl quinquefolius extract will also affect the effect of the composition. Cornus officinalis extract, Eleutherodactyl quinquefolius extract and Psidium guajava extract have a certain synergistic effect in antioxidant efficacy, and none of them can be missing.

[0170] Test Example 2

[0171] Collagenase activity inhibition assay

[0172] Experimental methods:

[0173] Use Tris-HCl buffer (pH = 7.5 ± 0.2, containing 5 mmol / mL CaCl2) to prepare 1 mg / mL tetracycline hydrochloride (positive control) and the sample solution to be tested; prepare 0.5 mg / mL substrate FALGPA buffer solution; prepare 0.2 mg / mL type I collagenase Tris-HCl solution.

[0174] Set up a blank group, a control group, a sample group, and a sample blank group. Add 140 μL of 0.2 mg / mL enzyme solution to the control group and the sample group, and add an equal volume of buffer to the blank group and the sample blank group. After 20 minutes of water bath at 37°C, add 40 μL of substrate FALGPA solution to each group. After 20 minutes of water bath at 37°C, measure the absorbance changes of each component at 330 nm. Each group is measured 3 times on average and the average value is taken. The inhibition rate of collagenase is calculated according to the following formula, and the results are shown in Table 2.

[0175] Sample blank group: sample solution 60μL + buffer salt 140μL + substrate 40μL

[0176] Sample group: sample solution 60μL + collagenase 140μL + substrate 40μL

[0177] Control group: 60 μL buffer salt + 140 μL collagenase + 40 μL substrate

[0178] Blank group: 200 μL buffer + 40 μL substrate

[0179]

[0180] Table 2

[0181]

[0182]

[0183] It can be seen from the data results in Table 2 that the composition described in the present application has a good collagenase inhibitory effect. The preparation method of Cornus officinalis extract and Eleutherodactyl quinquefolius extract will also affect the effect of the composition. Cornus officinalis extract, Eleutherodactyl quinquefolius extract and Psidium guajava extract have a certain synergistic effect in this effect, and none of them can be missing.

[0184] Test Example 3

[0185] AGEs inhibition assay

[0186] Experimental methods:

[0187] 1 mg / mL aminoguanidine hydrochloride (positive control) and the sample solution to be tested were prepared with phosphate buffer (50 mmol / L, pH 7.4). 1 mL of the sample to be tested, 0.8 mg / mL bovine serum albumin (BSA) solution, 200 mM glucose solution (GLU) and PBS buffer were added respectively, and heated continuously at 60°C for 24 hours. Fluorescent AGEs were determined by a fluorescent microplate reader with an excitation wavelength of 370 nm and an emission wavelength of 420 nm. The fluorescent AGEs content was expressed as fluorescence intensity AU. The fluorescence intensity of each group before and after the reaction was measured, and each group was measured 3 times on average to take the average value. The inhibition rate of the sample on the formation of fluorescent AGEs was calculated using the following formula, and the results are shown in Table 3.

[0188] Sample set: sample solution 1mL + buffer 1mL + BSA 1mL + GLU 1mL

[0189] Blank control group: PBS buffer 2mL + BSA 1mL + GLU 1mL

[0190]

[0191] Where:

[0192] A0 is the initial fluorescence intensity of the blank control group;

[0193] A1 is the initial fluorescence intensity of the experimental group;

[0194] B0 Fluorescence intensity of blank control group after reaction;

[0195] B1 Fluorescence intensity of the experimental group after reaction.

[0196] Table 3

[0197] Group IC50 value (μg / mL) Example 1 167.72 Example 2 178.64 Example 3 169.72 Example 4 203.93 Example 5 212.35 Example 6 226.80 Example 7 279.75 Example 8 208.32 Example 9 370.68 Example 10 324.09 Embodiment 11 305.47 Example 12 315.09 Comparative Example 1 394.56 Comparative Example 2 716.29 Comparative Example 3 535.2 Preparation Example 1-1 723.14 Preparation Example 2-1 285.49 Preparation Example 3-1 450.28 Positive Control 488.45

[0198] It can be seen from the data results in Table 3 that the composition described in the present application has a good inhibitory effect on the formation of fluorescent AGEs. The preparation method of Cornus officinalis extract and Eleutherodactyl quinquefolius extract will also affect the effect of the composition. Cornus officinalis extract, Eleutherodactyl quinquefolius extract and Psidium guajava extract have a certain synergistic effect in anti-glycation effect, and none of them can be missing.

[0199] Test Example 4

[0200] LPS-induced anti-inflammatory experiment of RAW264.7 macrophages

[0201] Test method:

[0202] According to the results of the CCK-8 method, the test concentration of each sample group was selected as 125 μg / mL to determine the anti-inflammatory efficacy of macrophage RAW264.7. A blank control group, a model group, a positive control group and a sample group were set up. Complete medium was added to each well of the blank control group, 5 μM dexamethasone sodium acetate diluted with complete medium was added to each well of the positive control group, and samples diluted with complete medium were added to each well of the sample group. Three replicate wells were set up for each group and incubated in a 37°C, 5% CO2 incubator for 2h. Except for the blank control group, LPS with a final concentration of 1 μg / mL was added to each well and incubated in a 37°C, 5% CO2 incubator for 24h. The cell supernatant was collected by centrifugation in a low-temperature centrifuge, and the NO detection kit was tested according to the instructions. The content of NO in the collected cell supernatant was detected, and the experimental data are shown in Table 4.

[0203] Sample group: samples diluted in complete medium and LPS;

[0204] Model group: complete medium and LPS;

[0205] Positive control group: 5 μM dexamethasone sodium acetate and LPS diluted in complete medium;

[0206] Blank control group: complete medium only.

[0207] Table 4

[0208]

[0209]

[0210] It can be seen from the data results in Table 4 that the composition described in the present application has a good inhibitory effect on the secretion of NO. The preparation method of Cornus officinalis extract and Eleutherodactyl quinquefolius extract will also affect the effect of the composition. Cornus officinalis extract, Eleutherodactyl quinquefolius extract and Psidium guajava extract have a certain synergistic effect in anti-inflammatory effect, and none of them can be missing.

[0211] Test Example 5

[0212] Zebrafish microcirculation improvement experiment

[0213] Zebrafish strain: wild-type AB strain, 30 fish per experimental group, cultured in a 28°C incubator. Experimental methods:

[0214] According to the results of MTC determination, a sample group with a concentration of 100 μg / mL was selected for the zebrafish microcirculation improvement experiment. 48hpf zebrafish with consistent developmental conditions were selected, and the zebrafish fry were randomly assigned to 6-well plates, with 30 per well. A single concentration group (100 μg / mL) was set in the sample group, and aspirin was selected as the positive drug. A normal control group and a model control group were also set up. Except for the normal control group, the remaining experimental groups were given 0.9% sodium chloride in water to establish a zebrafish microcirculatory disorder model, and then 3 mL of the test solution was added to each well. After incubation at 28°C for 24 hours, 10 zebrafish were randomly selected from each experimental group and placed under a blood flow analyzer to record zebrafish blood flow videos. The blood flow analyzer was used to analyze and process the mid-abdomen where the zebrafish blood vessels were the clearest and the blood flow was larger in the collected video, and the blood flow velocity (V) of the tested zebrafish was collected. The blood flow velocity of each group of zebrafish was analyzed and observed by the blood flow analyzer. The formula for improving the sample blood flow velocity is as follows, and the results are shown in Table 5.

[0215] Normal control group: containing zebrafish and standard diluted aqueous solution;

[0216] Model group: containing zebrafish prepared with 0.9% NaCl solution;

[0217] Sample group: Contains the test substance prepared with 0.9% NaCl solution and zebrafish.

[0218]

[0219] Table 5

[0220]

[0221] Note: Compared with the blank group, ### p<0.001, compared with the model group, * p<0.05, *** p<0.001

[0222] It can be seen from the data results in Table 5 that the composition described in the present application has a significant promoting effect on the blood flow rate of zebrafish. The preparation method of Cornus officinalis extract and Eleutherodactyla quinquefolia extract will also affect the effect of the composition. Cornus officinalis extract, Eleutherodactyla quinquefolia extract and Psidium guajava extract have a certain synergistic effect in improving microcirculation, and none of them can be missing.

[0223] The applicant declares that the present invention illustrates a composition with anti-inflammatory and anti-aging effects and its application through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0224] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0225] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A composition with anti-inflammatory and anti-aging effects, characterized in that: The composition includes cornus officinalis extract, quinquefolia extract and guava extract.

2. The composition with anti-inflammatory and anti-aging effects according to claim 1, characterized in that: The composition comprises, by weight, 10-50 parts of cornus officinalis extract, 20-60 parts of quinquefolia extract and 10-50 parts of guava extract.

3. The composition with anti-inflammatory and anti-aging effects according to claim 1, characterized in that: The cornus officinalis extract is prepared by a method comprising the following steps: (1) Mixing Cornus officinalis with water and performing ultra-high pressure microfluidization pretreatment; (2) mixing the pretreated Cornus officinalis with an enzyme for enzymolysis to obtain an enzymolysis solution; (3) The enzymatic hydrolyzate is filtered, concentrated, and dried to obtain the product.

4. The composition with anti-inflammatory and anti-aging effects according to claim 1, characterized in that: The five-fingered peach extract is prepared by a method comprising the following steps: (1) mixing the five-fingered peach with water, and extracting under ultrasound to obtain an extract; (2) Filter the extract, concentrate it, and dry it to obtain the product.

5. The composition with anti-inflammatory and anti-aging effects according to claim 1, characterized in that: The guava extract is prepared by a method comprising the following steps: (1) mixing guava with water and performing ultra-high pressure microfluidization pretreatment; (2) subjecting the pretreated guava to reflux extraction to obtain an extract; (3) The extract is centrifuged to obtain the supernatant, which is filtered, concentrated, and dried to obtain the product.

6. The composition with anti-inflammatory and anti-aging effects according to claim 3, characterized in that: The ultra-high pressure microfluidic pretreatment is performed under a pressure of 100-300 MPa, a temperature of 60-80° C., and for 2-6 times.

7. The composition with anti-inflammatory and anti-aging effects according to claim 3, characterized in that: The enzyme is a combination of cellulase and pectinase, and the added amount of the enzyme is 2-5% of the weight of the cornus officinalis; The enzymatic hydrolysis temperature is 35-55°C, the pH is 3-5.5, and the time is 20-30 minutes.

8. The composition with anti-inflammatory and anti-aging effects according to claim 4, characterized in that: Before the extraction, the method further comprises treating the mixture of the five-fingered peach and water at 110-130° C. and 0.1-0.2 MPa for 15-30 minutes.

9. The composition with anti-inflammatory and anti-aging effects according to claim 4, characterized in that: The temperature of the ultrasound is 60-80°C, the frequency is 30-40kHz, the time is 15-30min, and the number of times is 1-3 times.

10. Use of the composition with anti-inflammatory and anti-aging effects according to any one of claims 1 to 9 in the preparation of a product with anti-inflammatory, anti-aging, antioxidant or anti-glycation effects.

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