Edible and medicinal composition and application thereof in preparation of medicine for preventing and / or treating hyperuricemia

Abstract

The invention belongs to the technical field of traditional Chinese medicine compositions, and particularly relates to an edible and medicinal composition and application thereof in preparation of a medicine for preventing and / or treating hyperuricemia. According to the invention, five edible and medicinal components including the celery seed extract, the inonotus obliquus extract, the lycium ruthenicum extract, the honeysuckle flower extract and the corn stigma extract are compatible to obtain the edible and medicinal composition. Experiments prove that the edible and medicinal composition has relatively high antioxidant activity and xanthine oxidase inhibitory activity, can regulate the levels of inflammatory factors (IL-6, TNF-alpha, IL-10 and TGF-beta), regulate the content of malondialdehyde (MAD), regulate and improve the activity of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX), and can be used for preparing the food and medicinal composition. Therefore, the effect of reducing uric acid is achieved, and the hyperuricemia can be effectively prevented and / or treated.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine compositions, and in particular relates to a food-drug homologous composition and an application thereof in preparing a medicine for preventing and / or treating hyperuricemia. Background Art

[0002] Hyperuricemia (HUA) is a metabolic disease caused by excessive production or insufficient excretion of uric acid (UA) or both. Hyperuricemia is often accompanied by complications such as kidney damage, coronary heart disease, and hypertension. Therefore, the management of hyperuricemia is clinically significant in preventing diseases such as gout. The kidney is the main organ for UA excretion, and the kidney maintains the balance of UA in the body through filtration, reabsorption, and secretion. HUA usually triggers inflammation, which can lead to the production and accumulation of urate crystals in certain tissues and organs. These crystals can lead to the response of the immune system and promote the phagocytosis of urate by some cells such as macrophages. At the same time, these processes activate the expression of pro-inflammatory factors IL-6, TNF-α and anti-inflammatory factors IL-10, TGF-β, and activate inflammatory responses through some inflammatory signaling pathways (such as the NF-κB pathway), leading to an increase in the secretion of inflammatory cytokines.

[0003] Xanthine oxidase (XOD) continuously catalyzes the conversion of xanthine and hypoxanthine into uric acid. It is a key enzyme in the production of uric acid. By inhibiting its activity, the level of uric acid production can be reduced, providing an effective target for the prevention and management of gout. In addition, when XOD catalyzes the production of uric acid from xanthine, it consumes a large amount of oxygen molecules and produces reactive oxygen species (ROS). ROS are usually active in nature. Excessive accumulation of ROS can cause cellular oxidative damage, which can aggravate HUA. Therefore, anti-oxidation is also a top priority in the treatment of hyperuricemia. Some current drugs are used to manage HUA by inhibiting uric acid production (such as XOD inhibitors such as allopurinol) and promoting uric acid excretion (such as benzbromarone), but these drugs have serious side effects, including severe renal insufficiency, liver damage, gastrointestinal irritation, bone marrow suppression, and even secondary HUA. Safety and tolerability issues still exist. Therefore, it is necessary to find a safe, effective and economical drug to treat hyperuricemia.

[0004] my country is rich in food and medicine homologous substances, which have a positive impact on human health. Food and medicine homologous substances contain a variety of active ingredients, such as polyphenols, flavonoids, polysaccharides, alkaloids, etc. Some existing studies have also developed some combinations of food and medicine homologous substances for lowering uric acid, but the effect is not ideal. Summary of the invention

[0005] The object of the present invention is to provide a food-drug homologous composition and its use in the preparation of a medicament for preventing and / or treating hyperuricemia. The food-drug homologous composition has good uric acid-lowering activity and can achieve the effect of preventing and treating hyperuricemia.

[0006] The present invention provides a food-drug homologous composition, comprising the following components in parts by weight:

[0007] 38-44 parts of celery seed extract, 36-42 parts of inonotus birch extract, 8-12 parts of black wolfberry extract, 3-7 parts of honeysuckle extract and 3-7 parts of corn silk extract.

[0008] Preferably, based on the total weight parts of celery seed extract, inonotus obliquus extract, black wolfberry extract, honeysuckle extract and corn silk extract being 45 to 50 parts, the following components are also included in weight parts: 13 to 17 parts of kudzu root extract, 8 to 12 parts of Poria cocos extract, 3 to 5 parts of chicory extract, 1 to 2 parts of hawthorn extract and 1 to 2 parts of raspberry extract.

[0009] Preferably, the celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract are all water extracts;

[0010] The preparation method of the celery seed extract, honeysuckle extract or corn silk extract comprises the following steps: crushing the celery seed, honeysuckle or corn silk, mixing with water, sequentially performing a first soaking and a first reflux extraction, solid-liquid separation, and collecting a first extract; sequentially performing reduced pressure concentration and drying on the first extract to obtain the celery seed extract, honeysuckle extract or corn silk extract respectively;

[0011] The preparation method of the inonotus obliquus extract or the black wolfberry extract comprises the following steps: crushing the inonotus obliquus or the black wolfberry, mixing with water, sequentially performing a second soaking and a second reflux extraction, solid-liquid separation, and collecting a second extract; sequentially performing reduced pressure concentration and drying on the second extract to obtain the inonotus obliquus extract or the black wolfberry extract, respectively.

[0012] The present invention also provides the use of the food-drug homologous composition described in the above scheme in the preparation of uric acid-lowering products.

[0013] The present invention also provides the use of the food-drug homologous composition described in the above scheme in the preparation of a drug for preventing and / or treating hyperuricemia.

[0014] Preferably, the drug includes a drug having one or more functions of uric acid lowering activity, anti-inflammatory activity, antioxidant activity and xanthine oxidase inhibitory activity;

[0015] The anti-inflammatory effect includes one or more of the following four items: 1) reducing the production of pro-inflammatory factor IL-6; 2) reducing the production of pro-inflammatory factor TNF-α; 3) increasing the production of anti-inflammatory factor IL-10; 4) increasing the production of anti-inflammatory factor TGF-β;

[0016] The antioxidant activity includes one or more of the following four items: 1) reducing the content of malondialdehyde; 2) increasing the activity of catalase; 3) increasing the activity of superoxide dismutase; 4) increasing the activity of glutathione peroxidase.

[0017] Preferably, the effective concentration of the food-drug homologous composition in the medicine is 390.625-3125.00 μg / mL.

[0018] The present invention also provides the use of the food-drug homologous composition described in the above scheme in preparing a drug for preventing gout.

[0019] The present invention also provides a medicine for preventing and / or treating hyperuricemia, wherein the effective ingredients of the medicine include the food-drug homologous composition described in the above scheme.

[0020] The present invention also provides a compressed candy for lowering uric acid, comprising the following raw materials in parts by weight: 45 to 50 parts of celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract, 13 to 17 parts of kudzu root extract, 8 to 12 parts of poria extract, 3 to 5 parts of chicory extract, 1 to 2 parts of hawthorn extract, 1 to 2 parts of raspberry extract, 1 part of chitosan oligosaccharide, 13 to 17 parts of sorbitol and 1 part of magnesium stearate.

[0021] Beneficial effects:

[0022] The present invention provides a food-drug homologous composition, comprising the following components in parts by weight: 41 parts of celery seeds, 39 parts of inonotus birchii, 10 parts of black wolfberry, 5 parts of honeysuckle and 5 parts of corn silk. The present invention obtains the food-drug homologous composition by combining the above five food-drug homologous components. It has been found through experimental verification that the food-drug homologous composition has high antioxidant activity and xanthine oxidase inhibitory activity, and can also regulate the levels of inflammatory factors (IL-6, TNF-α, IL-10 and TGF-β), regulate the content of malondialdehyde (MAD), regulate and increase the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX), thereby achieving the effect of lowering uric acid, thereby effectively preventing and / or treating hyperuricemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0024] Figure 1 The scavenging ability of the food and medicine homologous substances on DPPH free radicals in Experimental Example 1;

[0025] Figure 2 The scavenging ability of the food and medicine homologous substances on ABTS free radicals in Experimental Example 1;

[0026] Figure 3 is the inhibition rate of the food and medicine homologous substances on xanthine oxidase in Experimental Example 2;

[0027] Figure 4 is XOI in FMHC1 during the in vitro digestion process in Example 5;

[0028] Figure 5 An example of a standard curve constructed by measuring the peak areas of purine standards at different concentrations (a) for the HPLC determination of purine bases in Example 6 and (b) for the HPLC chromatogram;

[0029] Figure 6 The effect of FMHC1 on HK-2 cell viability in Example 7;

[0030] Figure 7 The effect of FMHC1 on uric acid in hyperuricemia HK-2 cells in Example 7; wherein a is an HPLC graph of uric acid, inosine and adenosine in the cell supernatant, and b is a graph of uric acid levels in the cell supernatant;

[0031] Figure 8 The effect of FMHC1 on the production of inflammatory factors in hyperuricemia HK-2 cells in Example 8; wherein a is a graph showing IL-6 levels, b is a graph showing TNF-α, c is a graph showing IL-10 levels, and d is a graph showing TGF-β levels;

[0032] Fig. 9 The effect of FMHC1 on oxidative stress indicators of HK-2 cells in Example 9; wherein a is the MDA level graph, b is the CAT activity graph, c is the SOD activity graph, and b is the GSH-PX activity graph;

[0033] Fig.10 is the Chuanbei equation for FMHC2 powder in Example 11;

[0034] Fig.11 The effect of FMHC2 on HK-2 cell viability in Example 11;

[0035] Fig.12 The effect of FMHC2 on uric acid in hyperuricemia HK-2 cells in Example 11;

[0036] Fig.13The effect of FMHC2 on the production of inflammatory factors in hyperuricemia HK-2 cells in Example 11; wherein a is the level of IL-6; b is the level of TNF-α; c is the level of IL-10; and d is the level of TGF-β;

[0037] Fig.14 : The effect of FMHC2 on oxidative stress indicators of HK-2 cells in Example 11; wherein a is the MDA level graph, b is the CAT activity graph, c is the SOD activity graph, and b is the GSH-PX activity graph;

[0038] Fig.15 The storage stability index changes of FMHC2 in Example 11; wherein, a is the browning index; b is the sensory score; c is the total phenol content; and d is the total flavonoid content. DETAILED DESCRIPTION

[0039] The invention provides a food and medicine homologous composition, comprising the following components in parts by weight: 38-44 parts of celery seed extract, 36-42 parts of inonotus birch extract, 8-12 parts of black wolfberry extract, 3-7 parts of honeysuckle extract and 3-7 parts of corn silk extract.

[0040] As an embodiment, the food and medicine composition is composed of the following components: 38-44 parts of celery seed extract, 36-42 parts of inonotus birch extract, 8-12 parts of black wolfberry extract, 3-7 parts of honeysuckle extract and 3-7 parts of corn silk extract.

[0041] In parts by weight, the food and medicine composition of the present invention comprises 38 to 44 parts of celery seed extract; as an embodiment, the weight of the celery seed extract is 41 parts.

[0042] Based on the weight of celery seeds, the food and medicine composition of the present invention includes 36 to 42 parts of Inonotus obliquus extract; as an embodiment, the weight of the Inonotus obliquus extract is 39 parts.

[0043] Based on the weight of celery seeds, the food and medicine composition of the present invention includes 8 to 12 parts of black wolfberry extract; as an embodiment, the weight of the black wolfberry extract is 10 parts.

[0044] Based on the weight of celery seeds, the food and medicine composition of the present invention includes 3 to 7 parts of honeysuckle extract; as an embodiment, the weight of the honeysuckle extract is 5 parts.

[0045] Based on the weight of celery seeds, the food and medicine composition of the present invention includes 3 to 7 parts of corn silk extract; as an embodiment, the weight of the corn silk extract is 5 parts.

[0046] As an embodiment, in terms of weight percentage, the total amount of the celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract of the present invention is 100%. For example, in a specific embodiment of the present invention, the weight percentage of each component in the food and medicine composition is: 41% celery seed extract, 39% inonotus birch extract, 10% black wolfberry extract, 5% honeysuckle extract and 5% corn silk extract.

[0047] As an embodiment, based on the total weight parts of celery seed extract, inonotus obliquus extract, black wolfberry extract, honeysuckle extract and corn silk extract being 45 to 50 parts, the present invention also includes the following components in weight parts: 13 to 17 parts of kudzu root extract, 8 to 12 parts of poria extract, 3 to 5 parts of chicory extract, 1 to 2 parts of hawthorn extract and 1 to 2 parts of raspberry extract.

[0048] As an embodiment, the food and medicine composition is composed of the following components: 45 to 50 parts of celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract, 13 to 17 parts of kudzu root extract, 8 to 12 parts of Poria cocos extract, 3 to 5 parts of chicory extract, 1 to 2 parts of hawthorn extract and 1 to 2 parts of raspberry extract.

[0049] In the specific implementation of the present invention, the kudzu root extract, poria extract, chicory extract, hawthorn extract and raspberry extract are commercially available extracts purchased from Shaanxi Yuanbeibei Biotechnology Co., Ltd., and are commercially available extracts in a 10:1 ratio.

[0050] In the specific implementation of the present invention, the celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract are all water extracts.

[0051] In the specific implementation process of the present invention, the preparation method of the celery seed extract, honeysuckle extract or corn silk extract includes the following steps: crushing the celery seeds, honeysuckle or corn silk, mixing with water, sequentially performing a first soaking and a first reflux extraction, solid-liquid separation, and collecting a first extract; sequentially performing reduced pressure concentration and drying on the first extract to obtain celery seed extract, honeysuckle extract or corn silk extract, respectively. In one embodiment of the present invention, the mass ratio of the crushed celery seeds, honeysuckle or corn silk to water (solid-liquid ratio) is 1:(10-15), further 1:12; the water is pure water; the first soaking time is 2-4h, further 3h; the first reflux extraction is hot reflux extraction, and the temperature is 100°C; the first reflux extraction is performed 3 times, and the time of each first reflux extraction is 60-90min, and the extracts of the three first reflux extractions are combined; the reduced pressure concentration is to a solid content of 20%; the vacuum degree of the reduced pressure concentration is -0.085MPa; the temperature of the reduced pressure concentration is 50°C; the drying is spray drying; the inlet air temperature of the spray drying is 110-120°C.

[0052] In the specific implementation process of the present invention, the preparation method of the birch inonotus extract or the black wolfberry extract comprises the following steps: crushing the birch inonotus or the black wolfberry, mixing with water, sequentially performing a second soaking and a second reflux extraction, solid-liquid separation, and collecting the second extract; sequentially performing reduced pressure concentration and drying on the second extract to obtain the birch inonotus extract or the black wolfberry extract, respectively; in one embodiment of the present invention, the mass ratio of the material after the birch inonotus or the black wolfberry is crushed to water (solid-liquid ratio) is 1: (8-12), further 1: 10; the water is pure water; the second soaking time is 4 to 6 hours, further 5 hours; the second reflux extraction is hot reflux extraction, and the temperature is 100°C; the number of the second reflux extraction is 3 times, the time of each second reflux extraction is 90 to 120 minutes, and the extracts of the three second reflux extractions are combined; the reduced pressure concentration is to a solid content of 35% to 45%, further 40%; the vacuum degree of the reduced pressure concentration is -0.085MPa; the temperature of the reduced pressure concentration is 50°C; the drying is vacuum freeze drying.

[0053] In the present invention, the preparation method of the food-drug homologous composition comprises the following steps: mixing the celery seed extract, the inonotus birch extract, the black wolfberry extract, the honeysuckle extract and the corn silk extract to obtain the food-drug homologous composition; the mixing is based on uniform mixing.

[0054] In the present invention, the celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract have high antioxidant activity and xanthine oxidase inhibitory activity. After the five food and medicine homologous components are combined according to the components in the present invention, it is found that the food and medicine homologous composition can significantly reduce the increase of uric acid caused by hyperuricemia, and at the same time can regulate the levels of inflammatory factors IL-6, TNF-α, IL-10 and TGF-β, thereby achieving the effect of regulating the uric acid level of hyperuricemia and preventing and / or treating hyperuricemia.

[0055] Based on the above advantages, the present invention also provides the use of the food-drug homologous composition described in the above scheme in the preparation of a product for reducing uric acid. As an embodiment, the product includes food, health care products or medicines. As an embodiment, the uric acid reduction is to reduce the increase of uric acid caused by hyperuricemia.

[0056] The present invention also provides the use of the food-drug homologous composition described in the above scheme in the preparation of a drug for preventing and / or treating hyperuricemia. As an embodiment, the drug is a drug having one or more functions of uric acid lowering activity, anti-inflammatory activity, antioxidant activity and xanthine oxidase inhibitory activity; as another embodiment, the drug is a drug having uric acid lowering activity, anti-inflammatory activity, antioxidant activity and xanthine oxidase inhibitory activity.

[0057] As an embodiment, the anti-inflammatory includes one or more of the following four items: 1) reducing the production of proinflammatory factor IL-6; 2) reducing the production of proinflammatory factor TNF-α; 3) increasing the production of anti-inflammatory factor IL-10; 4) increasing the production of anti-inflammatory factor TGF-β.

[0058] As an embodiment, the antioxidant activity includes one or more of the following four items: 1) reducing the malondialdehyde content; 2) increasing the catalase activity; 3) increasing the superoxide dismutase activity; 4) increasing the glutathione peroxidase activity.

[0059] As one embodiment, the effective concentration of the food-drug homologous composition in the drug of the above technical solution is 390.625-3125.00 μg / mL; as another embodiment, the effective concentration of the food-drug homologous composition is 781.25-3125.00 μg / mL.

[0060] The present invention also provides the use of the food-drug homologous composition described in the above scheme in the preparation of a drug for preventing gout. The food-drug homologous composition of the present invention can further prevent gout complications caused by hyperuricemia through the prevention and / or treatment effect on hyperuricemia.

[0061] The present invention also provides a drug for preventing and / or treating hyperuricemia, wherein the active ingredient of the drug comprises the food-drug homologous composition described in the above scheme. As an embodiment, the drug further comprises a pharmaceutically acceptable excipient. The present invention does not specifically limit the type of the excipient, and it can be conventionally selected according to needs, such as according to the drug dosage form, or according to the drug preparation process.

[0062] The present invention also provides a compressed candy for lowering uric acid, comprising the following raw materials in parts by weight: 45 to 50 parts of celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract, 13 to 17 parts of kudzu root extract, 8 to 12 parts of poria extract, 3 to 5 parts of chicory extract, 1 to 2 parts of hawthorn extract, 1 to 2 parts of raspberry extract, 1 part of chitosan oligosaccharide, 13 to 17 parts of sorbitol and 1 part of magnesium stearate.

[0063] As an embodiment, the compressed candy is composed of the following raw materials in parts by weight: 45 to 50 parts of celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract, 13 to 17 parts of kudzu root extract, 8 to 12 parts of Poria cocos extract, 3 to 5 parts of chicory extract, 1 to 2 parts of hawthorn extract, 1 to 2 parts of raspberry extract, 1 part of chitosan oligosaccharide, 13 to 17 parts of sorbitol and 1 part of magnesium stearate.

[0064] As an embodiment, in terms of weight percentage, the weight percentage of each component in the compressed candy is: 20.5% celery seed extract, 19.5% inonotus birch extract, 5% black wolfberry extract, 2.5% honeysuckle extract, 2.5% corn silk extract, 15% kudzu root extract, 10% poria extract, 4% chicory extract, 2% hawthorn extract, 2% raspberry extract, 1% chitosan oligosaccharide, 15% sorbitol and 1% magnesium stearate.

[0065] The compressed candy of the present invention has excellent uric acid lowering effect and unique flavor, suitable sour and sweet taste, smooth and delicate taste, and also has good storage stability and is more suitable for storage at room temperature and under refrigeration conditions.

[0066] As an embodiment, the shape of the compressed candy includes oval, triangle, diamond or circle.

[0067] In the present invention, the preparation method of the tablet candy comprises the following steps: mixing the raw materials of the tablet candy, and then tableting to obtain the tablet candy; the tableting method is a direct tableting method.

[0068] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0069] The experimental materials, experimental reagents, main instruments and data processing methods used in the following experimental examples and embodiments are:

[0070] 1. Experimental Materials

[0071] The extracts of celery seeds, inonotus obliquus, black wolfberry, corn silk and honeysuckle were provided by the Bioengineering Laboratory of North University of China; the extracts of kudzu root, poria, chicory, hawthorn and raspberry were all commercially available extracts in a 10:1 ratio; chitosan oligosaccharide, food grade, was purchased from Shandong Weikan Biotechnology Co., Ltd.; sorbitol, microcrystalline cellulose and magnesium stearate were all food grade and purchased from Anhui Qianshun Biotechnology Co., Ltd.; renal tubular epithelial cells HK-2 were purchased from Wuhan Pronosai Life Science Technology Co., Ltd.

[0072] Experimental reagents

[0073] Folin (biotechnology grade), gallic acid (standard ≥ 98%), rutin (standard ≥ 98%), NaCO3, NaNO2, Al(NO3)3, NaNO2, NaOH, and anhydrous ethanol were all analytical grade, and all drugs were purchased from Shanghai McLean Biochemical Technology Co., Ltd.

[0074] Table 1 Other experimental reagents

[0075]

[0076]

[0077] 2. Main instruments

[0078] Table 2 Experimental instruments

[0079] Instrument Name Specifications Origin / Brand High Performance Liquid Chromatography (HPLC) ultimate3000 Thermo Fisher Scientific Analytical Balance ME204 / 02 Mettler-Toledo Instruments (Shanghai) Co., Ltd. <![CDATA[CO2 Incubator]]> MCO-18AC Panasonic Clean bench HFsafe-1500LC Shanghai Lishen Scientific Instrument Co., Ltd. Multifunctional ELISA reader ReadMax1500 Shanghai Shanpu Biotechnology Co., Ltd. Constant temperature incubator SPX-150B-Z Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory High-speed centrifuge TDL-5-A Shanghai Anting Scientific Instrument Factory Constant temperature water bath HH-6 Jiangsu Jintan Ronghua Instrument Manufacturing Co., Ltd. Magnetic stirrer SN-MS-3D Shanghai Shangpu Instrument Equipment Co., Ltd. Single punch tablet press DP-30 Shanghai Taberite Machinery Co., Ltd. Tablet Four-purpose Tester ZPJ-4 Tianjin Jingtuo Instrument Technology Co., Ltd. Centrifuge TDL-5-A Shanghai Anting Scientific Instrument Factory Analytical Balance ME-204 Mettler-Toledo Instruments (Shanghai) Co., Ltd. Electronic moisture meter DHS-20A Bangsi Instrument Technology (Shanghai) Co., Ltd.

[0080] 3. Data processing

[0081] The experimental data were expressed as the mean ± standard deviation of the results of three measurements; graphics were drawn using Origin 2022 software; uniform design data were statistically analyzed using DPS 7.05; other data were analyzed for differences using SPSS23.0 software, and P<0.05 indicated a significant statistical difference.

[0082] The Chinese, Latin and abbreviations of 11 kinds of food and medicine homologous substances used in the following experimental examples and embodiments are compared: celery seed (Apium graveolens L., AGL), birch inonotus (Inonotus obliquus, IO), black wolfberry (Lycium ruthenicum Murray, LRM), corn silk (Stigma Maydis, SM), honeysuckle (Lonicerajaponica Thunb, LJT), kudzu root (Pueraria lobata (Willd.) Ohwi, PLWO), Poria cocos (Poria cocos (Schw.) Wolf, PCSW), chicory (Cichorium intybus, CI), hawthorn (Crataegus pinnatifida Bunge, CPB) and raspberry (Rubus idaeus L., RIL) and chitosan oligosaccharide (Chitosan oligosaccharide, CO).

[0083] Experimental Example 1

[0084] Evaluation of Antioxidant Activity of Food and Drug Homologous Substances

[0085] The antioxidant activities of 11 edible and medicinal homologous substances were evaluated by DPPH and ABTS free radical inhibition tests. The DPPH free radical scavenging ability was determined according to the method in the following literature [Xiaojun Li, Yiyuan Fan, Juan Guo, et al. Polyvinylalcohol / kappa-carrageenan-based package film with simultaneous incorporation of ferric ions and polyphenols from Capsicum annuum leaves for fruit shelf-life extension [J]. International Journal of Biological Macromolecules, 2024, 266: 131002.] with slight modifications. Specifically, sample solutions (mother solutions of extracts of 11 food and drug homologous substances) with different mass concentrations (0.4, 0.8, 1.2, 1.6, 2.0 mg / mL) were prepared, 100 μL of sample solution and 100 μL of DPPH alcohol solution (0.2 mmoL / L) were respectively measured and mixed in a 96-well plate, and the absorbance value was measured at a wavelength of 517 nm after reacting at room temperature in the dark for 40 minutes. Vc was used as a positive control. The scavenging rate of the sample on DPPH free radicals was calculated according to formula (1).

[0086] DPPH free radical scavenging rate / % = [1-(A s -A r ) / A0]×100 (1)

[0087] Note: A s : Sample group (sample + DPPH alcohol solution); A r : Sample control group (sample + 95% ethanol); A0: blank control group (95% ethanol + DPPH alcohol solution)

[0088] The determination of ABTS free radical scavenging ability was slightly modified according to the method in the following literature [Zhang Jianfeng, Li Xiaojun, Wang Xiaohui, et al. Different extraction processes of hemp seed shell polyphenols and their antioxidant properties [J]. Food Research and Development, 2023, 44 (6): 153-158.]. Prepare sample solutions of different mass concentrations (0.10, 0.25, 0.40, 0.55, 0.70 mg / mL), measure 50 μL of sample solution and 150 μL of ABTS working solution (7 mmol / LABTS solution and 2.45 mmol / L potassium persulfate solution are mixed in equal volumes, reacted at room temperature in the dark for 12 to 16 hours to obtain ABTS stock solution, diluted with 70% ethanol solution to make its absorbance at 734 nm wavelength 0.7 ± 0.02) in a 96-well plate, reacted at room temperature in the dark for 6 minutes, and measured the absorbance at 734 nm. VC was used as a positive control. The scavenging rate of the sample for ABTS free radicals was calculated according to formula (2).

[0089] ABTS free radical scavenging rate / % = [1-(A i -A j ) / A0]×100 (2)

[0090] Note: A i : Sample group (sample + ABTS working solution); A j : Sample control group (sample + anhydrous ethanol); A0: blank control group (anhydrous ethanol + ABTS working solution)

[0091] Figures 1-2 Demonstrated the ability of different food and medicine homologous substances to clear DPPH and ABTS free radicals. Figure 1 It can be seen that within the experimental concentration range, as the sample concentration increases, the scavenging rates of the seven food and drug homologous substances, LRM, AGL, RIL, CPB, IO, SM and LJT, on DPPH free radicals gradually increase, showing a concentration-dependent manner, which is similar to the positive control VC (IC 50 =10.41±0.74μg / mL) showed the same DPPH free radical scavenging trend, and its IC 50The concentrations of CO, PCSW, CI and PLWO were 0.54±0.04, 0.93±0.02, 1.12±0.01, 1.26±0.01, 1.35±0.03, 1.4±0.03 and 1.58±0.02 mg / mL respectively (P<0.05), all of which showed strong DPPH free radical scavenging ability and had significant differences with VC (P<0.01). In addition, the scavenging rates of CO, PCSW, CI and PLWO on DPPH free radicals were weak. When the mass concentration reached 2 mg / mL, the scavenging rate of CO on DPPH free radicals was only 34.2±0.12%, and when the mass concentration reached 5 mg / mL, the scavenging rates of PCSW, CI and PLWO were almost zero.

[0092] Depend on Figure 2 It can be seen that with the increase of sample concentration, the inhibition rate of ABTS free radicals by seven edible and medicinal homologous substances, LRM, AGL, LJT, IO, SM, CPB and RIL, showed concentration dependence, which was consistent with the scavenging effect of DPPH free radicals. 50 The results showed that the ABTS free radical scavenging ability was good, but the scavenging effect was weaker than that of VC (IC 50 =7.85±0.03μg / mL). However, CO, PCSW, CI and PLWO have weak scavenging ability on ABTS free radicals. When the mass concentration reaches 2mg / mL, the scavenging rate of CO on DPPH free radicals is only 31.5%. When the mass concentration reaches 4mg / mL, PCSW, CI and PLWO almost have no scavenging effect. Therefore, the scavenging ability of the 11 food and drug homologous substances on ABTS free radicals is consistent with the scavenging ability of DPPH free radicals. Except for PCSW, CI, PLWO and CO, which have weak scavenging rates, other samples have good scavenging effects.

[0093] Experimental Example 2

[0094] Evaluation of Xanthine Oxidase (XOD or XO) Inhibitory Activity

[0095] Since the uric acid generated by XOD-catalyzed substrate xanthine has a characteristic absorption peak at a wavelength of 290 nm, the activity intensity of XO can be reflected by measuring the absorbance value at this wavelength within a certain period of time, and then the inhibitory effect of different samples on XO activity can be evaluated [refer to the following literature Kailin Chen, Dongdong Xie, Manping Luo, et al. Functional Food Potential of Chrysanthemum morifolium, Perilla frutescens, and Sophora japonica in Managing Hyperuricemia through Dual Enzyme Inhibition [J]. Journal of Agricultural and Food Chemistry, 2024, 72 (46): 25879-25894]. Sample solutions of different mass concentrations (2, 4, 6, 8, 10 mg / mL) were prepared, and 50 μL of phosphate buffer (PBS, pH=7.5), 70 μL of sample solution, and 50 μL of XOD solution (0.5 U / mL) were added to a 96-well plate in sequence. After incubation in a 37°C incubator for 10 min, 100 μL of xanthine solution (0.6 mmol / L) was added, and after reacting at 37°C for 20 min, the absorbance was measured at 290 nm. Allopurinol was used as a positive control. The XOD inhibition rate (XOI) was calculated according to formula (3).

[0096] XOI(%)=1-(A a -A b ) / (A c -A d ) (3)

[0097] Note: A a : Sample group (sample+XOD+PBS+xanthine); A b : Sample control group (sample+PBS+xanthine); A c : Blank group (XOD+PBS+xanthine); A d : Blank control group (PBS+xanthine).

[0098] XOD continuously catalyzes the conversion of xanthine and hypoxanthine into uric acid. The accumulation of uric acid in the body can lead to hyperuricemia. Therefore, XOD is a key enzyme in the production of uric acid. Inhibiting its activity to reduce uric acid levels can provide an effective target for the prevention and management of hyperuricemia. Figure 3As shown in the figure, it can be seen that with the increase of mass concentration, the XOI of LRM, LJT, IO, AGL, SM, CPB and RIL also gradually increased, showing a good dose-effect relationship, which was similar to the positive control AP (IC 50 =2.26±0.11μg / mL) showed the same XOI trend, IC 50 The inhibitory effects of CPB and RIL were weak, and the inhibitory effects of PCSW, CI, PLWO and CO on xanthine oxidase were not obvious. When the concentration reached 20 mg / mL, the inhibition rates were all less than 10%. Therefore, IO, LRM, AGL, LJT and SM were selected for the next test based on their antioxidant activity.

[0099] Experimental Example 3

[0100] Screening of uric acid-lowering food and medicine homologous compositions based on uniform design

[0101] Taking XOI as the evaluation index, the "uniform design method" was used to study the optimal compatibility of these five food-medicine homologous substances, in order to obtain a food-medicine homologous composite 1 (FMHC1) with higher activity.

[0102] Choose U 10 *(10 8 ) uniform design table (see Table 3), according to the uniform design table (see Table 4, group design, X1: black wolfberry, X2: honeysuckle, X3: corn silk, X4: birch inonotus, X5: celery seed. Referring to the results of the antioxidant test and XOD inhibition test, according to U 10 *(10 8 ) Using the instructions of the table, set the factors and levels, and optimize the uniform design scheme through DPS 7.05 software (see Table 5). Multivariate stepwise regression analysis was performed on the XOI of the 10 groups of food-drug homologous compositions to determine the optimal ratio and relationship between the food-drug homologous substances.

[0103] Table 3U 10 *(10 8 )

[0104]

[0105] Table 4U 10 *(10 8 ) usage table

[0106]

[0107] Table 5 Dosage table of food and drug homologous substances (10 mg / mL)

[0108] X1 X2 X3 X4 X5 N1 0.5 1.4 1.8 2.3 4.1 N2 0.6 1.8 2.4 3 2.1 N3 1.4 4.1 0.5 1.8 2.3 N4 1.8 0.5 2.3 4.1 1.4 N5 2.3 1.8 4.1 1.4 0.5 N6 1.8 2.1 0.6 2.4 3 N7 2.1 3 1.8 0.6 2.4 N8 2.4 0.6 3 2.1 1.8 N9 4.1 2.3 1.4 0.5 1.8 N10 3 2.4 2.1 1.8 0.6

[0109] The uniform design results are shown in Table 6. The results show that different doses of food and drug homologous substances combined in different proportions present a higher XOI, ranging from 77.15±2.13% to 86±1.25%, and the difference is statistically significant (P<0.05). Among them, the uniform design groups N2 and N6 are at a lower XOI level, the N1, N3, N9, and N10 groups are at a medium level, and there is no significant difference (P>0.05), while the N5 group has a higher XOI and is significantly different from other groups (except the N4 group) (P<0.05).

[0110] Table 6 XOI of uniform design group

[0111]

[0112] Note: Different letters indicate significant differences among the groups (P<0.05), the same below.

[0113] The significance test of partial regression coefficient can be used to determine the individual impact of the introduced fitting items on the dependent variable Y. As shown in Table 7, X3 (AGL), X4 (IO), X52 (SM), X1*X2 (LRM, LJT), X3*X5 (AGL, SM), and X4*X5 (IO, SM) all have significant impacts on XOI. The order of the individual variables affecting XOI is X4*X5>X1*X2>X3*X5>X52>X3>X4>X42>X1*X5, indicating that different food and drug homologous substances have a more significant impact on XOI when they interact with each other.

[0114] Table 7 t-test of partial regression coefficients

[0115]

[0116]

[0117] The experimental data were subjected to multivariate stepwise regression analysis, and the regression equation was obtained as Y = 82.5937 + 0.3343 * X3 + 0.5213 * X4-0.0093 * X42 + 0.0648 * X52-0.0495 * X1 * X2-0.0033 * X1 * X5-0.0739 * X3 * X5-0.1542 * X4 * X5, where X1, X2, X3, X4, and X5 represent LRM, LJT, AGL, IO, and SM, respectively. The determination coefficient R of the regression model 2=0.99995, significant level p=0.0151 (P<0.05), residual standard deviation S=0.0505, indicating that the regression model has good fitting and can be used to predict the XOI of the food-drug homologous composition at different dosage ratios. The optimal ratio X1:X2:X3:X4:X5 is 1:0.5:4.1:3.9:0.5, and the predicted value of XOI is 87.6109%. The optimal compatibility composition solution of 10 mg / mL was prepared for formula verification, and the measured value XOI was 87.1±0.79%, which was not significantly different from the predicted value (P<0.05), and was better than the XOI of the uniform design group N1~N10. Therefore, the optimal weight ratio of each substance in the food-drug homologous composition (FMHC1) optimized by the model is: AGL 41%, IO 39%, LRM 10%, LJT 5% and SM 5%.

[0118] Example 4

[0119] A food and medicine homologous composition (FMHC1) is composed of the following components in percentage by weight: 41% of celery seed extract, 39% of inonotus obliquus extract, 10% of black wolfberry extract, 5% of honeysuckle extract and 5% of corn silk extract.

[0120] The preparation method is:

[0121] 1. After celery seeds, honeysuckle and corn silk are crushed separately, pure water is added at a solid-liquid ratio of 1:12; soak for 3 hours; hot reflux extraction 3 times, solid-liquid separation, and the extracts are combined; reduced pressure concentration to a concentrate with a solid content of about 20%, the reduced pressure concentration conditions are vacuum degree -0.085MPa, temperature 50℃; spray drying to obtain the corresponding extracts.

[0122] 2. After the birch mushroom and black wolfberry are crushed separately, pure water is added at a solid-liquid ratio of 1:10; soak for 5 hours; perform hot reflux extraction 3 times, separate the solid and liquid, and combine the extracts; concentrate under reduced pressure to a concentrate with a solid content of 40%, the reduced pressure concentration conditions are a vacuum degree of -0.085MPa and a temperature of 50°C; vacuum freeze-dry to obtain the corresponding extracts.

[0123] 3. Weigh different extracts according to their weight percentages, mix them thoroughly and prepare a food-drug homologous composition, named FMHC1.

[0124] Experimental Example 5

[0125] In vitro saliva, gastric juice, and intestinal juice digestion simulation of FMHC1

[0126] (1) The preparation of electrolyte solution and saliva, gastric juice, and intestinal juice refers to the following literature:

[0127]

Mulet-Cabero AI, Egger L, Portmann R, et al. A standardised semi-dynamic in vitro digestion method suitable for food-an international consensus[J]. Food&function, 2020, 11(2): 1702-1720.

Zhu Miao. Study on the inhibitory effect of catechins on xanthine oxidase and the influence of processing conditions on its activity[D]. Nanchang University, 2022. DOI: 10.27232 / d.cnki.gnchu.2022.003761.

Liu Lian. Dissolution kinetics, simulated digestion and biological activity of polysaccharides from Porphyra yezoensis[D]. Jiangsu University, 2017.

[0128] Salivary fluid electrolytes (SSF): Accurately weigh 0.3822 g NaCl, 0.7455 g KCl, and 0.0666 g CaCl2, dissolve them in 500 mL distilled water, and then adjust the pH to 6.9 (± 0.05) with 1 mol / L HCl and 1 mol / L NaHCO3.

[0129] Simulated saliva (eSSF): Accurately weigh 0.493 g of α-amylase and dissolve it in 400 mL of SSF. Stir for 20 min and filter. Add 400 mL of SSF solution to the filtrate and mix well. Refrigerate for later use.

[0130] Gastric electrolytes (SGF): Accurately weigh 1.595 g NaCl, 0.55 g KCl, 0.075 g CaCl2 and 0.60 g NaHCO3, dissolve them in 500 mL distilled water, and then adjust the pH to 2.5 (± 0.05) with 1 mol / L HCl.

[0131] Simulated gastric fluid (eSGF): Accurately weigh 1.77 g of pepsin, 75 mg of gastric lipase and 1.5 mL of CH3COONa (pH 5.0, 1.0 mol / L) and add them to 150 mL of SGF. Stir magnetically at room temperature for 10 min, adjust the pH to 2.5 (±0.05) with 1 mol / L HCl, and refrigerate for later use.

[0132] Intestinal electrolytes (SIF): Accurately weigh 2.70 g NaCl, 0.325 g KCl, and 0.165 g CaCl2 and dissolve them in 500 mL distilled water, then adjust the pH to 7 (± 0.05) with 1 mol / L NaHCO3.

[0133] Simulated intestinal fluid (eSIF): Accurately weigh 13 mg of trypsin and add it to 100 mL of trypsin solution (7%, w / w, magnetic stirring for 10 min, centrifugation at 4000 rpm for 5 min, and supernatant), 200 mL of bile (4%, w / w) and 100 mL of SIF mixture, and then adjust the pH to 7.5 (±0.05) with 1 mol / L NaHCO3.

[0134] (2) In vitro digestion simulation method of FMHC1 obtained in Example 4

[0135] Simulated saliva digestion: 4 mL of FMHC1 solution (10 mg / mL) and 2 mL of LeSSF were added to a 50 mL centrifuge tube and mixed thoroughly. The mixture was reacted at 150 rpm in a 37°C constant temperature shaker for 5 min to simulate the dynamic digestion process. The enzyme was then quickly inactivated in a boiling water bath for 5 min to terminate the reaction. After cooling to room temperature, the digestion solution was used for the XOD inhibition test. The test method was the same as in Example 2, and each group was tested in parallel for 3 times. The test was divided into a sample group (sample + eSSF), a sample control group (sample + SSF), and a blank group (eSSF + distilled water).

[0136] Simulated saliva-gastric juice continuous digestion: Take a 150mL conical flask, add 20mL of FMHC1 solution (10mg / mL) and 20mL LeSSF, mix well and react at 150rpm at 37℃ constant temperature shaker for 5min, quickly boil in water for 5min to inactivate enzyme activity, adjust pH of digestive solution after saliva digestion to 2.5 with 1mol / L HCI, mix with 20mL eSGF, and adjust pH to 2.5 with 1mol / L HCI to obtain mixed digestive solution. Take 4 50ml centrifuge tubes in each group, add 15mL of the above mixed digestive solution, mix well and react at 150rpm at 37℃ constant temperature shaker. After reacting for 0, 1, 2 and 4h respectively, take out the corresponding centrifuge tubes and boil in water for 5min to inactivate enzyme to terminate the reaction. After cooling at room temperature, use digestive solution to perform XOD inhibition test, the test method is the same as Example 2, and each group is parallel 3 times. The experiment was divided into a sample group (sample + eSSF + eSGF), a sample control group (sample + SSF + SGF), and a blank group (eSSF + eSGF + distilled water).

[0137] Simulated saliva-gastric juice-intestinal juice continuous digestion: Take a 150mL conical flask, add 20mL of FMHC1 solution (10mg / mL) and 20mL of eSSF, mix well, react at 150rpm in a constant temperature shaker at 37℃ for 5min, quickly boil in water for 5min to inactivate enzyme activity, then mix with 20mL of LeSGF, adjust pH to 2.5 with 1mol / L HCl, mix well, react at 150rpm in a constant temperature shaker at 37℃ for 4h for the second stage of gastric digestion, then quickly boil in water for 5min to inactivate enzyme activity, and adjust pH to 7 with 1mol / L NaHCO3. After digestion with saliva and gastric juice, mix with 20ml of eSIF, adjust pH to 7 with 1mol / L NaHCO3 to obtain a mixed digestion solution. Take 4 50mL centrifuge tubes in each group, add 20mL of the above mixed digestion solution respectively, mix well, react at 150rpm in a constant temperature shaker at 37℃. After the above experiments were reacted for 0, 1.5, 1 and 2 hours, the corresponding centrifuge tubes were taken out and boiled in water for 5 minutes to kill the enzyme to terminate the reaction. After cooling to room temperature, they were placed in a refrigerator for storage, and each group was paralleled 3 times. After cooling to room temperature, the XOD inhibition test was carried out with the digestive juice, and the test method was the same as in Example 2, and each group was paralleled 3 times. The test was divided into a sample group (sample + eSSF + eSGF + eSIF), a sample control group (sample + SSF + SGF + SIF), and a blank group (eSSF + eSGF + eSIF + distilled water).

[0138] After digestion simulation, the in vitro xanthine oxidase inhibition rate (XOI) of FMHC1 was as follows Figure 4As shown in the figure, its XOI decreased significantly (P<0.05), among which the XOI of the blank group did not change significantly after digestion, but there was still a low XOI, which may be due to the influence of digestive enzymes and pH on the metabolic process of XOD catalyzing xanthine to produce uric acid, thereby affecting XOI [Reference: Ding Qiao DQ, Nie ShaoPing NSP, Hu JieLun HJL, et al. In vitro and invivo gastrointestinal digestion and fermentation of thepolysaccharide fromGanoderma atrum[J].2017.]. In addition, compared with the sample control group, the XOI of FMHC1 after simulated oral digestion only changed slightly, and there was no significant difference with the sample control group and the initial group (P>0.05). After 2 hours of simulated oral-gastric continuous digestion, XOI began to decrease. Compared with the sample control group, the relative XOI decreased by 0.72±0.82% (P>0.05), 1.52±0.98% (P>0.05), 3.77±1.34% (P<0.05), and 5.61±1.02% (P<0.01) after continuous digestion of gastric juice for 0, 1, 2, and 4 hours, respectively. During the entire simulated oral-gastric-intestinal continuous digestion stage, XOI continued to decrease during the intestinal juice digestion stage. Compared with the sample control group, the relative XOI decreased by 7.12±1.05% (P<0.01), 8.8±0.59% (P<0.01), 11.5±1.21% (P<0.01), and 18.2±1.55% (P<0.01) after continuous digestion of intestinal juice for 0, 0.5, 1, and 2 hours, respectively. The reason why FMHC1's XOI decreases after digestion may be that it contains a large number of active substances such as polyphenols and flavonoids. A large number of hydroxyl groups in these active substances interact with digestive enzymes (α-amylase, pepsin, gastric lipase, pancreatin, trypsin) through hydrogen bonds to form macromolecular polymers, resulting in a decrease in active substances after digestion and a decrease in XOI. In contrast, the relationship between the degree of decrease in XOI of FMHC1 at each digestion stage is intestine > stomach > oral cavity. The reason may be that the oral digestion process is short and does not cause loss of active substances. FMHC1 is a weakly acidic composition and is relatively stable in the acidic environment of simulated gastric fluid, which can protect active substances. However, it is easy to degrade and oxidize in the weakly alkaline intestinal fluid, so XOI is significantly reduced.

[0139] In summary, the presence of digestive enzymes and the pH value of digestive fluid have a great influence on the digestive stability of FMHC1 in the mouth, stomach, and intestines, which is consistent with the results of simulated digestion of polyphenols (such as caffeic acid, chlorogenic acid, gallic acid, etc.) and flavonoids (apigenin, quercetin, catechins, etc.) compounds in vitro [Reference: Zhu Miao. Study on the inhibitory effect of catechins on xanthine oxidase and the influence of processing conditions on its activity [D]. Nanchang University, 2022. DOI: 10.27232 / d.cnki.gnchu.2022.003761.].

[0140] Experimental Example 6

[0141] HPLC analysis

[0142] Preparation of standard solution: Accurately weigh 6 mg of adenosine, 6 mg of inosine, 3 mg of hypoxanthine, 4 mg of xanthine, 4 mg of adenine and 4 mg of uric acid, respectively, add a small amount of 0.10 mol / L NaOH to assist dissolution, and finally adjust the volume to 2.40 mL to prepare a 100 μmol / L standard solution, and store it at 4°C for later use.

[0143] High performance liquid chromatography can accurately analyze the changes and expression levels of different secondary metabolites, and a two-phase mobile phase was selected for gradient elution. Mobile phase A is 0.1% phosphoric acid (w / v), and mobile phase B is pure acetonitrile solution. The procedure is as follows: 0-15min, 2%-95% B; 15-25min, 95%-2% B. The total flow rate is set to 1.0mL / min. Before injecting the first sample, the corresponding system should be balanced at a given flow rate for at least 20min to ensure the stability of the column pressure. The injection volume is 20μL, the column temperature is 30℃, and the references are as follows: [Lin S, Meng J, Li F, et al. Ganoderma lucidum polysaccharide peptide alleviates hyperuricemia by regulating adenosinedeaminase and urate transporters[J]. Food&Function, 2022, 13(24): 12619-12631.].

[0144] The compounds in FMHC1 were determined using isocratic elution with a mobile phase of acetonitrile:0.01% acetic acid (w / v) = 40:60, a total flow rate of 1.0 mL / min, an injection volume of 20 μL, and a column temperature of 25°C.

[0145] Determination of purine bases by HPLC

[0146] Purines are divided into exogenous (derived from diet) and endogenous (derived from the degradation of DNA and RNA), and uric acid is the end product of purine metabolism in the human body [Zhang Y, Chen JS, Wang MM, et al. Synthesis and bioactivity evaluation of novel nuciferine derivatives with antihyperuricemia and nephroprotective effects [J]. Bioorganic Chemistry, 2022, 126: 105916.]. In order to construct a high-quality hyperuricemia cell model, HPLC was used to quantitatively determine different purine metabolites in the culture supernatant, including uric acid and its related precursors. Six standard samples were prepared and analyzed by HPLC. The results are as follows Figure 5 As shown, purine bases including inosine, adenosine, uric acid, xanthine, hypoxanthine and adenine are clearly separated. The peak area (Y), concentration (X) and determination coefficient (R2) were obtained according to the chromatographic conditions. The concentration range of uric acid, hypoxanthine, xanthine and adenine was 6.25-200 mg / L, which produced a strong linear relationship. The linear relationship of adenosine and inosine in the concentration range of 11-347 mg / L also had good reproducibility. In summary, the selected HPLC operating parameters are suitable for this study.

[0147] Experimental Example 7

[0148] (1) Detection of HK-2 cell viability by FMHC1

[0149] CCK-8 cell viability assay

[0150] HK-2 cells were seeded into 96-well plates at a density of 6000 cells / well, and 100 μL of culture medium containing different concentrations of FMHC (FMHC1: 48.83, 97.67, 195.31, 390.63, 781.25, 1562.50, 3125.00, 6250.00 μg / mL) was added for 48 h. The old culture medium was discarded, and 100 μL of culture medium containing 10% CCK-8 solution was added to each well. A culture medium without 10% CCK-8 solution was set as a blank control. The cells were incubated in an incubator at 37°C and 5% CO2 for 2 h, and then the OD value of each well was detected at a wavelength of 450 nm using an ELISA reader. The cell survival rate was calculated.

[0151] Cell viability (%) = [A(drug addition) - A(blank)] / [A(0 drug addition) - A(blank)] × 100;

[0152] A (drug added): absorbance of wells with cells, CCK-8 solution, and drug solution;

[0153] A (blank): absorbance of wells with culture medium and CCK-8 solution but no cells;

[0154] A (0 drug addition): absorbance of the wells with cells, CCK-8 solution but no drug solution.

[0155] In order to screen the concentration of HK-2 cell culture, different concentrations of FMHC1 (48.83, 97.67, 195.31, 390.63, 781.25, 1562.50, 3125.00, 6250.00 μg / mL) were used to act on the cells for 48 h, and then the cell viability was detected using CCK-8 reagent. Figure 6 As shown, different concentrations of FMHC1 have different tolerance to cells. With the increase of FMHC1 concentration, HK-2 cell viability first increases and then decreases. When the concentration reaches 1562.5 μg / mL, the cell survival rate is the highest, showing good cell tolerance. Therefore, three concentrations of FMHC1 (781.25, 1562.50, 3125.00 μg / mL) were selected as the next low, medium, and high dosing concentrations.

[0156] (2) Effect of FMHC1 on hyperuricemia HK-2 cells

[0157] Hyperuricemia HK-2 cell model

[0158] Cell culture: HK-2 cells were cultured in RPMI 1640 medium containing 10% FBS (fetal bovine serum, 100 μg / mL streptomycin, and 100 units / mL penicillin). The cells were cultured at 37°C in a humidified atmosphere supplemented with 5% CO2, and the culture medium was replaced every other day [Hou C, Liu D, Wang M, et al. Novel xanthine oxidase-based cell model using HK-2 cell for screening antihyperuricemic functional compounds [J]. Free Radical Biology and Medicine, 2019, 136: 135-145.].

[0159] Establishment of HK-2 cell model of hyperuricemia: HK-2 cells were cultured in complete medium to sub-confluence, and blank control group (no addition), model group (2.5mmol / L adenosine + 0.5U / mLXOD), positive control group (2.5mmol / L adenosine + 0.5U / mL XOD + 1mmol / L allopurinol); high-concentration sample group (2.5mmol / L adenosine + 0.5U / mL XOD + 3125μg / mLFMHC1), medium-concentration sample group (2.5mmol / L adenosine + 0.5U / mLXOD + 1563μg / mLFMHC1), and low-concentration sample group (2.5mmol / L adenosine + 0.5U / mLXOD + 781μg / mLFMHC1) were set up. The cells were seeded in a 6-well plate at a density of 300,000 cells / well (120,000 cells / mL) and incubated at 37°C for 48 hours. The blank control group and the model group were replaced with new culture medium. The positive control group and the sample group were pre-cultured for 24 hours, and then the culture medium was aspirated and washed three times with PBS. Adenosine in 2.5 mmol / L serum-free culture medium was added to the model group, the positive control group and the sample group, respectively. The blank control group was maintained in a fresh culture medium without adenosine. After incubation for 30 hours, 0.5 U / mL XOD was added to each well. After treatment for 8 hours, the culture supernatant was collected and the uric acid content was determined by HPLC to determine whether the model was successfully constructed. The references are as follows: [Lv SM, Zhang MQ, Chen JS, et al. Study on the anti-hyperuricemic bioactivity and chemical components of Sterculiae lychnophorae Semen [J]. Journal of Functional Foods, 2022, 95: 105173.].

[0160] The HK-2 model of hyperuricemia induced by exogenous adenosine and XOD was established to verify the effect of FMHC1 on the uric acid content in the cell supernatant. Figure 7 As shown in Figure a, HPLC was used to detect UA, adenosine and inosine in the cell culture supernatant. Figure 7 As shown in middle b, the uric acid content in the cell culture supernatant of the model group was significantly higher than that of the blank group (P<0.05), indicating that the hyperuricemia HK-2 cell model was successfully constructed. Compared with the model group, allopurinol in the positive control group significantly reduced the uric acid content of hyperuricemia HK-2 cells, and the low, medium and high dose groups of FMHC1 showed the same therapeutic effect, among which the medium dose group (1562.5μg / mL) had the most significant therapeutic effect, which may be related to the higher cell viability of this dose group. The results show that the stimulation of FMHC1 may have a certain protective effect on hyperuricemia HK-2 cells.

[0161] Experimental Example 8

[0162] Effect of FMHC1 on the levels of inflammatory factors in hyperuricemia cells

[0163] After the cells were administered, the crushed cells were resuspended in commercial lysis buffer and incubated at 4°C for 30 min. After low-temperature centrifugation (12000r / min, 20min), the supernatant was collected for pro-inflammatory cytokine detection. The concentrations of IL-6, TNF-α, IL-10 and TGF-β were determined using ELISA kits, and each group was repeated three times.

[0164] Adenosine and XOD induce hyperuricemia in HK-2 cells, which release pro-inflammatory and anti-inflammatory factors such as IL-6, TNF-α, IL-10 and TGF-β. These inflammatory factors are small secretory proteins that regulate inflammation, which can stimulate, recruit and amplify immune cells, initiate immune responses, and thus produce inflammation [Reference: Yang B, Xin M, Liang S, et al. Naringenin ameliorates hyperuricemia by regulating renal uric acid excretion via the PI3K / AKT signaling pathway and renal inflammation through the NF-κB signaling pathway [J]. Journal of Agricultural and Food Chemistry, 2022, 71(3): 1434-1446.]. In order to determine the inhibitory effect of FMHC1 on pro-inflammatory factors and the promoting effect of anti-inflammatory factors in hyperuricemia HK-2 cells, this example studied its effects on IL-6, TNF-α, IL-10 and TGF-β in hyperuricemia HK-2 cells by enzyme immunoassay (ELISA). Figure 8 As shown, the expression of IL-6 (P < 0.01) and TNF-α (P < 0.01) in the model group increased significantly, and the expression of IL-10 (P < 0.01) and TGF-β (P < 0.01) decreased significantly, indicating that hyperuricemia does induce cell inflammation. In addition, treatment with FMHC1 reduced the production of pro-inflammatory factors IL-6 (P < 0.05) and TNF-α (P < 0.01), increased the production of anti-inflammatory factors IL-10 (P < 0.05) and TGF-β (P < 0.05), and showed the same therapeutic effect as allopurinol, indicating that FMHC1 can reduce inflammation caused by hyperuricemia.

[0165] Experimental Example 9

[0166] Effect of FMHC1 on oxidative stress response in hyperuricemia cells

[0167] After the cells were dosed, the crushed cells were resuspended in a commercial lysis buffer and incubated at 4°C for 30 min. After low-temperature centrifugation (12000r / min, 20min), the supernatant was collected for detection of oxidative stress kinases and substances. CAT, GSP-Px, SOD, and MDA were detected using the BCA method, and the operation was performed according to the instructions of the ELISA kit, and each group was repeated three times.

[0168] Hyperuricemia can lead to oxidative stress response in the body, reduce the activity of antioxidant enzymes such as CAT, SOD, GSH-PX, and increase the level of MDA. This example explored the effect of FMHC1 on various oxidative stress indicators in HK-2 cells by enzyme immunoassay (ELISA). The results are as follows: Fig. 9 As shown. Fig. 9 As can be seen in a, the MDA level in the model group was significantly higher than that in the control group (P < 0.05), indicating that HUA promoted the production of MDA and successfully induced the oxidative stress response of cells. After administration of FMHC1 at different doses, the MDA level was significantly reduced (P < 0.05). Fig. 9 As can be seen in Figure b, the CAT activity in the model group was low, and the therapeutic effects of FMHC1 and allopurinol in the high-dose group were comparable (P>0.05). Fig. 9 As can be seen in Figure c, compared with the control group, the SOD activity in the model group was significantly reduced (P < 0.05), but after administration, the SOD activity increased, and the SOD activity in the medium-dose group was significantly better than that in the allopurinol group (P < 0.05). Fig. 9 As can be seen in Figure d, compared with the model group, the high-dose group significantly increased the level of GSH-PX (P < 0.05). HUA can induce oxidative stress in cells, but after administration of FMHC1 in different dose groups, the MD level of cells was reduced and the activity of antioxidant enzymes was increased, indicating that FMHC1 can effectively alleviate the oxidative stress caused by HUA.

[0169] Example 10

[0170] The tableted candy is prepared by direct tableting method using celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract, corn silk extract, kudzu root extract, poria extract, chicory extract, hawthorn extract and raspberry extract as main raw materials, with auxiliary materials such as chitosan oligosaccharide, sorbitol and magnesium stearate added.

[0171] Formula: In terms of mass percentage, it contains 20.5% celery seed extract, 19.5% inonotus birch extract, 5% black wolfberry extract, 2.5% honeysuckle extract, 2.5% corn silk extract, 15% kudzu root extract, 10% poria extract, 4% chicory extract, 2% hawthorn extract, 2% raspberry extract, 1% chitosan oligosaccharide, 15% sorbitol and 1% magnesium stearate.

[0172] Process flow: raw material pretreatment → preparation of raw material extract → adding a certain proportion of auxiliary materials → mixing → tableting → quality inspection → packaging → finished product, the finished product is named FMHC2.

[0173] Embodiment 11

[0174] The optimal formula of compressed candy was determined using sensory evaluation and friability as evaluation indicators, and the product quality was evaluated by physical and chemical indicators, microbiological indicators, etc. The uric acid-lowering effect of the optimal formula of compressed candy was verified by establishing a hyperuricemia HK-2 cell model. In addition, the storage stability of compressed candy was evaluated using browning index, sensory evaluation, changes in total phenol content, and changes in total flavonoids content as indicators.

[0175] 1. Determination and optimization of tablet candy formula

[0176] The optimal addition range of several main ingredients is obtained through the "chessboard method", and the main ingredient ratio, auxiliary material type, mold shape, etc. are adjusted for tableting. Sensory evaluation and brittleness are used as evaluation indicators to obtain the optimal formula, which is set as food and drug homologous composition 2 (FMHC2). Subsequent quality index testing and in vitro activity determination are carried out to determine the formula of tablet candy with uric acid lowering function. The mold shapes include ellipse, triangle, diamond, and circle.

[0177] 2. Determination of quality indicators of tablet candy

[0178] 1) Sensory evaluation

[0179] Strictly follow the requirements and methods of sensory evaluation, select 15 highly sensitive professional tasters to form an evaluation team. After observing the status of the compressed candy products, sensory evaluation was conducted from four aspects: taste (25 points), flavor (25 points), appearance (25 points), and chewiness (25 points). The evaluation criteria are shown in Table 8.

[0180] Table 8 Evaluation criteria for tablet candies

[0181]

[0182]

[0183] 2) Tablet weight difference

[0184] According to the Chinese Pharmacopoeia, 20 tablets of compressed candy are accurately weighed for total weight. After the average tablet weight is obtained, each tablet is accurately weighed. The weight of each tablet is compared with the average tablet weight. If the weight of a single tablet is less than 0.30g, the weight ratio is required to be less than ±7.5%; if the weight of a single tablet is greater than or equal to 0.30g, the weight ratio is required to be less than ±5%.

[0185] 3) Friability

[0186] According to the "Tablet Friability Test Method" of the Chinese Pharmacopoeia, take about 6.5g of compressed candy, set the test parameters to 21°C, 25r / min, and 4min, and use a tablet four-purpose tester for testing [China Pharmacopoeia Committee. Pharmacopoeia of the People's Republic of China - Part I: 2020 Edition [M]. Beijing: China Medical Science and Technology Press, 2020.]. Calculate the friability of the compressed candy according to formula (3-1), and the weight loss shall not exceed 1%.

[0187] Friability (%) = [(m1-m2) / m1] × 100% (3-1)

[0188] In formula (3-1), m1 is the total weight of the compressed candy before the test (g), and m2 is the total weight of the compressed candy after the test (g).

[0189] 4) Disintegration time

[0190] Refer to the Chinese Pharmacopoeia, use the tablet four-in-one instrument, take 6 tablets of candy and place them in the disintegration tank, set the temperature to 37°C, observe the disintegration time of the candy, and record it. According to the Chinese Pharmacopoeia 0921 disintegration time limit test method, each tablet should be completely disintegrated within 15 minutes.

[0191] 5) Moisture content

[0192] According to the provisions of my country's "National Food Safety Standard" GB / T5009.3-2016, the direct drying method is used to measure the moisture content of the material. About 5g of raw materials are weighed, and the raw materials are placed in the tray of the electronic moisture meter. The raw materials are heated from room temperature to 100°C, and dried at 100°C until the material has a constant weight. The instrument reading is read and the moisture content of the material is recorded.

[0193] 6) Kawakita Equation

[0194] For reference, use a funnel to slowly and evenly inject the prepared powder into a 500 mL measuring cylinder until the loose volume is about 200 mL. Allow the measuring cylinder filled with powder to fall freely from a height of 2 cm above the horizontal tabletop. Record the number of drops (n) and the corresponding volume (Vn mL), and process the data according to the Kawakita equation (3-2), (3-3), and (3-4).

[0195] n / C=1 / ab+n / a (3-2)

[0196] a=(v0-vinf) / v0 (3-3)

[0197] C=(v0-vn) / v0 (3-4)

[0198] In the formula, C is the relative volume reduction fraction of the powder; n is the number of taps, that is, the number of drops; a is the relative volume reduction fraction (the final volume reduction number) when the number of taps is infinite. The smaller a is, the better the fluidity of the powder; b is the filling rate constant. The larger b is, the better the filling property of the powder. The calculation of a and b adopts the drawing method, and n / C is drawn against n. The slope of the straight line is 1 / a, and the intercept is 1 / ab.

[0199] 7) Carr Index

[0200] Weigh 100g of sample into a measuring cylinder, tap until the sample height no longer changes, and record the sample volume before and after tapping. According to formulas (3-5), (3-6), and (3-7), the bulk density ρb (g / mL), tap density ρt (g / mL), and Carr's index c are processed respectively.

[0201] ρb=100 / vb (3-5)

[0202] ρt=100 / vt (3-6)

[0203] c=(1-ρb / ρt)×100% (3-7)

[0204] Wherein, Vb is the volume before compaction (mL), Vt is the volume after compaction (mL), and Carr's index c refers to the ratio of the difference between the tap density and the bulk density of the same sample to the tap density. If the value is between 15% and 25%, it indicates that the powder has good fluidity.

[0205] 8) Material bulk density

[0206] During the test, the material is loaded into a 500mL measuring cylinder and the measuring cylinder is vibrated until the material gap approaches the limit. When the material volume no longer decreases, the material volume at this time is recorded and the weight of the measuring cylinder before and after the material is added is weighed. Each material is measured three times in parallel and the bulk density of the material is calculated according to formula (3-8).

[0207] Bulk density ρ = (m2-m1) / v (3-8)

[0208] In formula (3-8), m2 is the total weight of the material and the measuring cylinder, m1 is the weight of the measuring cylinder, and v is the volume of the material.

[0209] 9) Angle of repose

[0210] The injection method (funnel fixing method) is used to evaluate the fluidity and friction of the material. The funnel is fixed at a position 12 cm away from the horizontal base plate, and 150 mL of compressed candy powder material is slowly injected to allow it to fall freely to form a cone. After the accumulation is stable, the cone height h and bottom radius r are measured, and the repose angle θ is calculated using formula (3-9).

[0211] θ=Arctan(h / r) (3-9)

[0212] The smaller the angle of repose, the better the fluidity. When it is less than 40°, the powder has good fluidity and can meet the fluidity requirements during tableting. When it is greater than 40°, it is necessary to modify the particle surface or add auxiliary materials to improve the fluidity of the powder.

[0213] 10) Color difference analysis

[0214] Use a colorimeter to measure the L*, a*, b*, and ΔE of the compressed candy. L* represents the brightness of the sample, a* represents the red-green value, b* represents the yellow-blue value, and ΔE represents the color difference. Each sample is measured three times and the average value is calculated.

[0215] 11) Microbiological indicators

[0216] The total colony count and Escherichia coli are tested in accordance with GB 17399-2016 "National Food Safety Standard Candy", GB 4789.2-2022 "National Food Safety Standard Food Microbiology Examination Total Colony Count Determination" and GB 4789.3-2022 "National Food Safety Standard Food Microbiology Examination Coliform Count".

[0217] 12) Nutritional composition determination

[0218] In accordance with GB 28050-2011 "National Food Safety Standard General Rules for Nutrition Labeling of Pre-packaged Foods", with reference to GB5009.5-2016 "National Food Safety Standard Determination of Protein in Foods", GB 5009.6-2016 "National Food Safety Standard Determination of Fat in Foods", GB 5009.7-2016 "National Food Safety Standard Determination of Reducing Sugars in Foods", and GB5009.91-2017 "National Food Safety Standard Determination of Potassium and Sodium in Foods", the protein (Kjeldahl nitrogen method), fat (Soxhlet extraction method), carbohydrates (reducing sugar titration method) and sodium (flame atomic absorption spectrometry) in compressed candies were determined.

[0219] 3. Establishment of HK-2 cell model of hyperuricemia

[0220] Same as Example 7.

[0221] 4. CCK-8 cell viability assay

[0222] The concentration gradient of FMHC2 is 48.83, 97.67, 195.31, 390.63, 781.25, 1562.50, 3125.00, 6250.00, 12500 μg / mL, and the specific method is the same as Example 7.

[0223] 5. Effect of FMHC2 on the levels of inflammatory factors in hyperuricemia cells

[0224] Same as Example 8.

[0225] 6. Effect of FMHC2 on oxidative stress response in hyperuricemia cells

[0226] Same as Example 9.

[0227] 7. Storage stability of compressed candy

[0228] In order to evaluate the storage stability of compressed candies, the compressed candies were stored at 4°C, 25°C, and 50°C for 4 weeks, and the browning index, sensory evaluation, total phenolic content, and total flavonoids content were evaluated every week.

[0229] 1) Browning index

[0230] The browning index was calculated by measuring the L*, a* and b* of the compressed candy using a colorimeter. The browning index was calculated according to formula (3-10).

[0231] Browning index = 100 (X-0.31) / 0.172 (3-10)

[0232] Where: X = (a* + 1.75L*) / (5.645L* + a* - 3.012b*)

[0233] 2) Determination of total phenol content

[0234] Determination by Folin-Ciocalteu method. Weigh 10 mg of gallic acid, dissolve it in water, and dilute it to a 100 mL volumetric flask. Take 0.1, 0.2, 0.3, 0.4, 0.5, and 0.6 mL of gallic acid and dissolve them in a 10 mL volumetric flask, add 0.3 mL of Folin-Ciocalteu solution and shake well. After 1 min, add 1.5 mL of Na2CO3 (20%), dilute to 10 mL, shake well and incubate at room temperature for 1 h. After the reaction is completed, measure the absorbance at 760 nm to complete the standard curve determination. Prepare 20 mg / mL FMHC2 solution, take 0.1 mL, add 0.3 mL of Folin-Ciocalteu solution and shake well. After 1 min, add 1.5 mL of Na2CO3 (20%), dilute to 10 mL, shake well and incubate at room temperature for 1 h. After the reaction is completed, measure the absorbance at 760 nm, and use deionized water as a blank control. The total phenolic content was expressed as gallic acid equivalent per gram of sample (μgGAE / g).

[0235] 3) Determination of total flavonoid content

[0236] The total flavonoids in the compressed candy were determined by the NaNO2-Al(NO3)3 colorimetric method. 10, 20, 30, 40, 50, 60 μg / mL rutin solutions were prepared, 1 mL of rutin solutions of different concentrations or 20 mg / mL FMHC2 solution was taken, 1 mL of 70% ethanol was added, 0.3 mL of 5% NaNO2 solution was added, shaken, and allowed to stand for 6 min, 0.3 mL of 10% Al(NO3)3 solution was added, shaken, allowed to stand for 6 min, and 2 mL of 4% NaOH was added. After standing for 10 min, the absorbance was measured at 510 nm. A standard curve was drawn with rutin, 70% ethanol was used as a blank control, and the total flavonoids were expressed as rutin equivalent (mgRE / g).

[0237] 8. Data Analysis

[0238] The experimental data were expressed as the mean ± standard deviation of the results of three measurements; graphics were drawn using Origin 2022 software; other data were analyzed for differences using SPSS23.0 software, and p<0.05 indicated a significant statistical difference.

[0239] 9. Results and Discussion

[0240] 1) Formula determination and optimization of compressed candy

[0241] The “chessboard method” was used to adjust the addition amount of FMHC1 and auxiliary materials such as sorbitol, microcrystalline cellulose, and magnesium stearate, and the shape of the tablet press mold was changed to optimize the performance of the compressed candy. Taking brittleness and sensory evaluation as indicators, it was finally determined that the addition amount of FMHC1 was 50%, the mold shape was round (diameter 12 mm), and the optimal formula FMHC2 for compressed candy is shown in Table 9.

[0242] Table 9 Optimal formula for tablet candy (FMHC2)

[0243] name FMHC1 PLWO PCSW CI CPB RIL CO Sorbitol Magnesium Stearate Addition amount (%) 50 15 10 4 2 2 1 15 1

[0244] Note: FMHC1 is the food-drug homologous composition prepared in Example 4, PLWO is Pueraria extract, PCSW is Poria extract, CI is Chicory extract, CPB is Hawthorn extract, RIL is Raspberry extract, and CO is Chitosan oligosaccharide.

[0245] The brittleness, tablet weight difference, and disintegration time of FMHC2 tablet candy all meet the standards. The surface of the tablet candy is smooth and complete, the taste is delicate, and there is no granularity. It has a unique caramel flavor, slightly bitter, and a suitable sweet and sour taste. The sensory score reached 89.8±0.65 points. The formula was further verified by experiments, and the experiment was repeated three times. The results showed that the quality indicators of the tablet candy all met the standards, and the sensory evaluation reached 90.1±0.80 points, which was consistent with the experimental results.

[0246] 2) Quality evaluation of compressed candy

[0247] The sensory evaluation of FMHC2 was carried out and the results are shown in Table 10.

[0248] Table 10 Sensory evaluation of FMHC2

[0249]

[0250] The physical and chemical indicators and microbiological indicators of FMHC2 were tested and all were within the qualified range. The results are shown in Table 11.

[0251] Table 11 Physical and chemical indicators and microbiological indicators of FMHC2

[0252]

[0253] Powder fluidity study Based on the experimental data, the powder index of FMHC2 was plotted with n / C as the ordinate and n as the abscissa. Fig.10 It can be seen that the regression equation of FMHC2 powder is Y = 3.3882X + 29.281 (R 2 =0.9807), a=0.2951, b=0.1157, the Chuanbei equation of FMHC2 is n / C=n / 0.2951+29.281. From the values ​​of a and b, it can be seen that the powder of FMHC2 has good fluidity and filling properties.

[0254] According to the experimental data, the Carr index of FMHC2 powder is 20.45%, which is between 15% and 25%, indicating that the powder has good fluidity, which is consistent with the result of the Chuanbei equation. In addition, from the results of bulk density and repose angle, it can be concluded that FMHC2 powder also has good stacking properties, and the FMHC2 color difference analysis shows that the product color is brown. Specific experimental data are shown in Table 12.

[0255] Table 12 Fluidity index and color difference analysis of FMHC2

[0256]

[0257] Nutrition label The nutrition label design of FMHC2 is shown in Table 13.

[0258] Table 13 Nutrition label of FMHC2

[0259] project Per 100g NRV% energy 1630 kilojoules (KJ) 19％ protein 1.8 grams (g) 3％ Fat 0 grams (g) 0％ carbohydrate 94.1 grams (g) 31％ sodium 26 milligrams (mg) 1％

[0260] 3) In vitro activity verification of compressed candies

[0261] The effect of FMHC2 on HK-2 cell viability was detected by using CCK-8 reagent to screen the culture concentration of FMHC2 acting on HK-2 cells. The results are as follows Fig.11 As shown in the figure, its action pattern is similar to that of FMHC1. When the concentration reaches 3125 μg / mL, the cell survival rate is the highest, showing good cell tolerance. Therefore, three concentrations of FMHC2 (1563, 3125, 6250 μg / mL) were selected as the subsequent low, medium, and high dosing concentrations. The effect of FMHC2 on hyperuricemia HK-2 cells is shown in the figure. Fig.12 As shown in a, HPLC was used to detect UA, adenosine and inosine in the cell culture supernatant. Fig.12 As shown in b, the uric acid content in the model group was significantly higher than that in the blank group (P<0.05), indicating that the hyperuricemia HK-2 cell model was successfully constructed. Compared with the model group, the low, medium and high dose groups of FMHC2 all showed good uric acid lowering effects, among which the therapeutic effects of the medium dose group (3125μg / mL) and the high dose group (6250μg / mL) had no significant difference (P>0.05), while the positive control group showed similar therapeutic effects.

[0262] The effect of FMHC2 on the levels of inflammatory factors in hyperuricemia cells was studied by ELISA. Fig.13 As shown in the results, compared with the blank group, the expression of anti-inflammatory factors IL-6 (P < 0.01) and TNF-α (P < 0.01) in the model group was significantly increased, and the expression of pro-inflammatory factors IL-10 (P < 0.01) and TGF-β (P < 0.01) was significantly decreased, indicating that hyperuricemia induced inflammation in the cell model. In addition, compared with the model group, treatment with high, medium, and low doses of FMHC2 reduced the production of IL-6 (P < 0.05) and TNF-α (P < 0.01), increased the production of anti-inflammatory factors IL-10 (P < 0.05) and TGF-β (P < 0.05), and all showed similar therapeutic effects to allopurinol, indicating that FMHC2 can regulate the disordered levels of inflammatory factors caused by hyperuricemia, thereby alleviating inflammation.

[0263] The effect of FMHC2 on oxidative stress indices in hyperuricemia cells was studied by ELISA. Fig.14 As shown in the results, the MDA level in the model group was significantly higher than that in the control group (P < 0.05). After administration of different doses of FMHC2, the MDA level was significantly reduced (P < 0.05), and the FMHC2 reduction effect in the medium and high dose groups was significant, indicating that FMHC2 can effectively alleviate cellular lipid peroxidation; the CAT, SOD and GSH-PX activities in the model group were significantly lower than those in the drug administration group (P < 0.05), indicating that the model group reduced the activity of antioxidant enzymes and induced oxidative stress response, but after the action of different doses of FMHC2, the levels of CAT, SOD and GSH-PX were significantly increased (P < 0.05), indicating that FMHC2 can effectively improve the oxidative stress caused by hyperuricemia.

[0264] In summary, HK-2 cells can maintain good cell viability at higher concentrations of FMHC2, and FMHC2 can significantly reduce the uric acid level in the supernatant of hyperuricemia cells, while regulating the levels of inflammatory factors and oxidative stress indicators in cells, effectively relieving inflammation and oxidative stress, and showing similar therapeutic effects as FMHC1. This result shows that even after physical optimization of the tablet candy preparation process, the therapeutic effect of the active substances of edible and medicinal materials on hyperuricemia has not been weakened, that is, the product shows good uric acid-lowering ability and inflammation-relieving ability. This conclusion provides more possibilities for edible and medicinal materials in the field of uric acid-lowering product development.

[0265] 4) Storage stability evaluation of compressed candies

[0266] During the storage process, the sensory coefficient of compressed candy will change with the change of time, temperature and humidity, and the bioactive substances such as polyphenols and flavonoids will be degraded to a certain extent. Therefore, the browning index, sensory score, change in total flavonoid content, and change in total polyphenol content are measured at different temperatures within a certain period of time to evaluate its storage stability. Fig.15 As shown in a, the browning index of compressed candy is relatively stable when stored at 4℃ and 25℃, but it increases by 10.5 when stored at 50℃ for 4 weeks, indicating that compressed candy is more likely to brown at high temperature. Fig.15 As shown in Figure b, temperature and time changes did not have a significant effect on the sensory evaluation of compressed candies. After one month of storage, the contents of total phenols and total flavonoids decreased to a certain extent. Fig.15 c and Fig.15The total phenols decreased by 400, 500, and 995 μgGAE / g at 4℃, 25℃, and 50℃, respectively, and the total flavonoids decreased by 500, 530, and 841 μgRE / g at 4℃, 25℃, and 50℃, respectively. The results showed that the active substances such as total phenols and total flavonoids in compressed candies were more easily degraded at high temperatures, while the degradation was slower under refrigeration and room temperature. In summary, the browning index and sensory evaluation of compressed candies were relatively stable during storage, while the active substances such as total phenols and total flavonoids would change to varying degrees, and compressed candies were more suitable for storage at room temperature and refrigeration to maximize the drug-dose effect.

[0267] 10. Conclusion

[0268] By optimizing the addition amount of 10 raw materials with the same origin as food and medicine, the addition amount of different auxiliary materials and the shape of the tablet press mold, a tablet candy product with unique flavor, appropriate sweet and sour, smooth and delicate texture was prepared with brittleness and sensory evaluation as evaluation indicators. The specific formula is 20.5% celery seed extract, 19.5% birch inonotus extract, 5% black wolfberry extract, 2.5% honeysuckle extract, 2.5% corn silk extract, 15% kudzu root extract, 10% Poria cocos extract, 4% chicory extract, 2% hawthorn extract, 2% raspberry extract, 1% chitosan oligosaccharide, 15% sorbitol and 1% magnesium stearate. The product quality was evaluated, and its sensory evaluation, physical and chemical indicators and microbiological indicators, powder fluidity and filling properties all met the standards. The uric acid-lowering effect of compressed candy was verified by establishing a hyperuricemia HK-2 cell model induced by adenosine and XOD. The results showed that after the compressed candy preparation process, its uric acid-lowering effect was similar to that of FMHC1, and it could still effectively reduce the uric acid level in the supernatant of hyperuricemia cells and relieve inflammation. It is a uric acid-lowering product with both flavor and activity. Finally, the storage stability of compressed candy was investigated in combination with the browning index, sensory evaluation, total phenol content and total flavonoid content. The results showed that compressed candy has good storage stability and is more suitable for storage at room temperature and refrigerated conditions.

[0269] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A food-drug homologous composition, characterized in that: The composition comprises the following components in parts by weight: 38-44 parts of celery seed extract, 36-42 parts of inonotus obliquus extract, 8-12 parts of black wolfberry extract, 3-7 parts of honeysuckle extract and 3-7 parts of corn silk extract.

2. The food-drug homologous composition according to claim 1, characterized in that: Based on the total weight parts of celery seed extract, inonotus obliquus extract, black wolfberry extract, honeysuckle extract and corn silk extract being 45 to 50 parts, the following components are also included in weight parts: 13 to 17 parts of kudzu root extract, 8 to 12 parts of poria extract, 3 to 5 parts of chicory extract, 1 to 2 parts of hawthorn extract and 1 to 2 parts of raspberry extract.

3. The food-drug homologous composition according to claim 1, characterized in that: The celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract are all water extracts; The preparation method of the celery seed extract, honeysuckle extract or corn silk extract comprises the following steps: The celery seeds, honeysuckle or corn silk are crushed, mixed with water, and sequentially subjected to a first soaking and a first reflux extraction, solid-liquid separation, and a first extract is collected; the first extract is sequentially subjected to reduced pressure concentration and drying to obtain a celery seed extract, a honeysuckle extract or a corn silk extract, respectively; The preparation method of the Inonotus obliquus extract or the black wolfberry extract comprises the following steps: The inonotus obliquus or black wolfberry is crushed, mixed with water, and sequentially subjected to a second soaking and a second reflux extraction, solid-liquid separation, and a second extract is collected; the second extract is sequentially subjected to reduced pressure concentration and drying to obtain an inonotus obliquus extract or a black wolfberry extract, respectively.

4. Use of the food-drug homologous composition according to any one of claims 1 to 3 in the preparation of a product for lowering uric acid.

5. Use of the food-drug combination according to any one of claims 1 to 3 in the preparation of a drug for preventing and / or treating hyperuricemia.

6. The use according to claim 5, characterized in that: The drugs include drugs having one or more functions of uric acid lowering activity, anti-inflammatory activity, antioxidant activity and xanthine oxidase inhibitory activity; The anti-inflammatory effect includes one or more of the following four items: 1) reducing the production of pro-inflammatory factor IL-6; 2) reducing the production of pro-inflammatory factor TNF-α; 3) increasing the production of anti-inflammatory factor IL-10; 4) increasing the production of anti-inflammatory factor TGF-β; The antioxidant activity includes one or more of the following four items: 1) reducing the content of malondialdehyde; 2) increasing the activity of catalase; 3) increasing the activity of superoxide dismutase; 4) increasing the activity of glutathione peroxidase.

7. The use according to claim 5 or 6, characterized in that: The effective concentration of the food-drug homologous composition in the medicine is 390.625-3125.00 μg / mL.

8. Use of the food-drug combination according to any one of claims 1 to 3 in the preparation of a drug for preventing gout.

9. A drug for preventing and / or treating hyperuricemia, characterized in that: The active ingredients of the medicine include the food-drug homologous composition according to any one of claims 1 to 3.

10. A compressed candy for lowering uric acid, characterized in that: The invention comprises the following raw materials in parts by weight: 45-50 parts of celery seed extract, inonotus birch extract, black wolfberry extract, honeysuckle extract and corn silk extract, 13-17 parts of kudzu root extract, 8-12 parts of poria extract, 3-5 parts of chicory extract, 1-2 parts of hawthorn extract, 1-2 parts of raspberry extract, 1 part of chitosan oligosaccharide, 13-17 parts of sorbitol and 1 part of magnesium stearate.

Images