Application of composition in preparation of iron supplementing preparation

Abstract

The invention discloses application of a composition in preparation of an iron supplementing preparation. The composition comprises dogwood and / or a dogwood extract and mangosteen and / or a mangosteen extract. Through compatibility of the active components, the composition provided by the invention can promote absorption of an iron element by a body by promoting intestinal iron absorption and / or inhibiting hepcidin and other mechanisms, plays a role in supplementing iron, further can be used for treating indications such as functional iron deficiency and the like, provides a more effective preparation for improving iron deficiency of the body, and has wide application prospects. Meanwhile, the preparation prepared from the composition also has the effects of regulating physique, tonifying deficiency and qi, regulating the health condition of the body and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine preparations, and in particular to application of a composition in the preparation of an iron supplement preparation. Background Art

[0002] From single-cell bacteria to multicellular organisms such as humans, iron is one of the most important metals for maintaining life. Iron plays a vital role in several cellular processes such as DNA synthesis, nucleic acid repair, mitochondrial cell respiration, cell growth and cell death, and contributes to host defense and cell signaling. Most of the iron in the human body is associated with red blood cell hemoglobin. Iron deficiency can lead to anemia, which in turn leads to a series of health problems, such as insufficient Qi and blood, cold hands and feet, and other symptoms.

[0003] Iron absorbed by enterocytes can be directly used in the cell's intrinsic metabolic machinery, stored in ferritin, or released through the basal outer membrane for systemic iron delivery. Most of the iron absorbed in the body is obtained from food, and Fe 3+ Must first be reduced to Fe 2+ Iron can only be absorbed by the intestines. The iron absorbed by the intestines is transported to the liver for storage. Hepcidin released by the liver is a key regulator of iron homeostasis. It binds to FPN, the only known cellular iron export protein, mediates the ubiquitination, internalization and degradation of FPN, and directly blocks the channel, thereby blocking the export of iron from cells to plasma. When serum iron levels increase, hepcidin expression is upregulated, resulting in the obstruction of iron transport to plasma through FPN, thereby providing a negative feedback reaction to prevent potential toxic iron accumulation in the body. Reduced plasma iron levels lead to reduced transferrin saturation, resulting in reduced hepcidin synthesis. Therefore, Hepcidin expression is enhanced, downregulating the expression of ferroportin and reducing the body's absorption of iron. When the body is iron deficient, the above process undergoes the opposite changes, thereby maintaining iron homeostasis.

[0004] Therefore, in order to improve a series of adverse effects caused by iron deficiency in the body, it becomes more important to explore safe and effective bioactive substances for iron supplementation. Summary of the invention

[0005] Based on this, it is necessary to provide a composition with excellent iron supplementation effect for use in the preparation of iron supplementation preparations.

[0006] In a first aspect, the present invention provides an application of a composition in the preparation of an iron supplement preparation, wherein the composition comprises ingredients of Cornus officinalis and Mangosteen, wherein the Cornus officinalis includes Cornus officinalis medicinal materials and / or Cornus officinalis extracts, and the mangosteen includes Mangosteen medicinal materials and / or Mangosteen extracts.

[0007] In some embodiments, the composition satisfies at least one of the following (1) to (3):

[0008] (1) The mass ratio of Cornus officinalis to Mangosteen in the composition is (10-20): (1-5);

[0009] (2) The Cornus officinalis extract is selected from a water extract and / or an alcohol extract;

[0010] (3) The mangosteen extract is selected from water extract and / or alcohol extract.

[0011] In some embodiments, the composition further comprises 2'-methoxymatrine and / or pyralidinol A.

[0012] In some embodiments, the composition satisfies the following (1) and / or (2):

[0013] (1) The amount of 2'-methoxymatrine added to the composition is 0.1-1 μM;

[0014] (2) The amount of Panlongol A added to the composition is 0.05~0.5 μM.

[0015] In some embodiments, the composition comprises 10-20 μg / mL Cornus officinalis extract, 1-5 μg / mL Mangosteen extract, 0.1-1 μM 2'-methoxymatrine and 0.05-0.5 μM Penicillin A.

[0016] In some embodiments, the iron supplement preparation achieves the iron supplement effect by promoting intestinal iron absorption and / or inhibiting hepcidin.

[0017] In a second aspect, the present invention also provides a complex, the components of which include Cornus officinalis and Mangosteen, the Cornus officinalis includes Cornus officinalis medicinal materials and / or Cornus officinalis extracts, the mangosteen includes Mangosteen medicinal materials and / or Mangosteen extracts, and also contains 2'-methoxymatrine and / or panlongol A.

[0018] In some embodiments, the composite satisfies at least one of the following (1) to (3):

[0019] (1) The content ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM;

[0020] (2) The mass ratio of Cornus officinalis, Mangosteen and Panlongphenol A in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.05-0.5) μM;

[0021] (3) The mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlongol A in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM: (0.05-0.5) μM.

[0022] In some embodiments, the compound satisfies at least one of the following (1) to (5):

[0023] (1) The mass ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the composite is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM;

[0024] (2) The mass ratio of Cornus officinalis, Mangosteen and Panlongphenol A in the composite is (15-18) μg / mL: (3-4) μg / mL: (0.05-0.2) μM;

[0025] (3) The mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlongol A in the composite is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM: (0.05-0.2) μM;

[0026] (4) The Cornus officinalis extract is selected from a water extract and / or an alcohol extract;

[0027] (5) The mangosteen extract is selected from water extract and / or alcohol extract.

[0028] In some embodiments, the composition of the complex includes 10-20 μg / mL Cornus officinalis extract, 1-5 μg / mL Mangosteen extract, 0.1-1 μM 2'-methoxymatrine and 0.05-0.5 μM Penicillin A.

[0029] In a third aspect, the present invention also provides a drug for treating anemia, wherein the drug comprises the complex and pharmaceutically acceptable excipients.

[0030] In some embodiments, the drug satisfies at least one of the following (1) to (4):

[0031] (1) The drug is in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes or aerosols;

[0032] (2) The dosage form of the drug includes oral preparation, injection, patch, ointment, spray, inhalation, drops, lotion or suppository;

[0033] (3) The anemia is iron deficiency anemia, autoimmune hemolytic anemia or nausea anemia;

[0034] (4) The drug is used to treat anemia by promoting intestinal iron absorption and / or inhibiting hepcidin.

[0035] In a fourth aspect, the present invention further provides a health product for replenishing qi and blood, the ingredients of the health product comprising the complex and auxiliary materials.

[0036] In some embodiments, the health care product is in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes or aerosols.

[0037] In a fifth aspect, the present invention also provides a multifunctional food for improving sub-health status, wherein the ingredients of the food include the complex and food additives.

[0038] Compared with the prior art, the present invention has the following beneficial effects:

[0039] The present invention makes full use of the characteristics of oral medicine and food homologous substances, and based on the advantage that food can be directly orally taken, combines the Chinese medicinal raw materials of Cornus officinalis and mangosteen and / or the active ingredients extracted therefrom, so that the obtained composition has a better ability to promote iron absorption, significantly improves the body's absorption efficiency of iron, plays an iron supplementing role, and can promote intestinal iron absorption and / or inhibit hepcidin, so that the composition can be used to prepare a drug for treating anemia indications, which can be used to treat anemia indications such as iron deficiency anemia, autoimmune hemolytic anemia or nausea anemia. At the same time, the drug prepared by using the composition also has significant effects of regulating physical constitution, replenishing deficiency and invigorating qi, regulating the health status of the body, etc., and has broader application prospects in the fields of health care products, food, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] FIG. 1 shows the effects of each single substance and combination formulas 1 to 6 in Experimental Example 1 on the survival rate of Caco-2 cells.

[0041] FIG. 2 shows the effects of each single substance and combination formulas 1 to 6 in Experimental Example 1 on the survival rate of HepG2 cells.

[0042] FIG. 3 is a statistical data of the effects of each single substance and combination formulas 1 to 6 in Experimental Example 2 on the absorption of iron ions by Caco-2 cells.

[0043] FIG. 4 is a statistical data of the effects of each single substance and combination formulas 1 to 6 in Experimental Example 3 on the mRNA expression of Hepcidin gene in HepG2 cells.

[0044] Figure 5 The data of each single substance and combination formulas 1 to 6 in Experimental Example 4 on the expression level of Hepcidin protein in HepG2 cells are statistically shown. DETAILED DESCRIPTION

[0045] In order to facilitate the understanding of the present invention, the present invention is described more fully below. Preferred embodiments of the present invention are provided, but the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0047] It should be noted that the experimental methods in the following examples of the present invention without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturers. The various commonly used chemical reagents used in the examples are all commercially available products.

[0048] Cornus officinalis (scientific name: Cornus officinalis Sieb. et Zucc.), also known as cornus meat, yam meat, and cornus meat, is the dried mature pulp of Cornus officinalis Sieb. et Zucc., a plant of the Cornaceae family. It tastes sour and astringent, is slightly warm in nature, and enters the liver and kidney meridians. It has the effects of nourishing the liver and kidneys, astringing and consolidating.

[0049] Mangosteen, (scientific name: Garcinia mangostana L.), also known as: mangosteen, or mountain bamboo, mangosteen, mangosteen. It is flat in nature, sweet and slightly sour. Mangosteen peel: cool in nature, bitter and astringent, invigorates the spleen and produces body fluid, and stops diarrhea. Mangosteen peel: anti-inflammatory and analgesic, mainly used to treat spleen deficiency diarrhea, dry mouth, burns, scalds, eczema, and stomatitis.

[0050] In a first aspect, the present invention provides an application of a composition in the preparation of an iron supplement preparation, wherein the composition comprises ingredients of Cornus officinalis and Mangosteen, wherein the Cornus officinalis includes Cornus officinalis medicinal materials and / or Cornus officinalis extracts, and the mangosteen includes Mangosteen medicinal materials and / or Mangosteen extracts.

[0051] The present invention combines the above-mentioned ingredients and finds that the multiple ingredients in each formula can synergistically promote iron absorption, thereby playing the role of iron supplementation for the body.

[0052] In some embodiments, the content ratio of Cornus officinalis to Mangosteen in the composition is (10-20): (1-5), including but not limited to 10:1, 10:2, 10:3, 10:4, 10:5, 12:1, 14:1, 16:1, 18:1, 18.2:1, 20:1, 12:2, 14:2, 16:2, 18:2, 20:2, 12:3, 14:3, 15:3, 16:3.2, 18:3, 20:3, 12:4, 14:4, 15:4, 16:4, 18:4, 20:4, 12:5, 14:5, 14.2:5, 16:5, 18:5, 20:5 or any range formed by any two of the foregoing and any ratio within the range.

[0053] In some embodiments, the Cornus officinalis medicinal material includes the pulp of Cornus officinalis, the peel of Cornus officinalis, the flower of Cornus officinalis, the bark of Cornus officinalis and other parts.

[0054] In some embodiments, the cornus officinalis extract includes substances extracted from the pulp, peel, flower, bark, etc. of cornus officinalis;

[0055] Optionally, the cornus officinalis extract includes substances such as alkaloids, polysaccharides, saponins, flavonoids and polyphenols.

[0056] In some embodiments, the Cornus officinalis extract is selected from a water extract and / or an alcohol extract.

[0057] In some embodiments, the mangosteen medicinal material includes mangosteen pulp, mangosteen peel, mangosteen flower, mangosteen bark root, stem and other parts.

[0058] In some embodiments, the mangosteen extract includes substances extracted from mangosteen pulp, mangosteen peel, mangosteen flower, mangosteen bark, root, stem and other parts;

[0059] Optionally, the mangosteen extract includes substances such as alkaloids, polysaccharides, saponins, flavonoids and polyphenols.

[0060] In some embodiments, the mangosteen extract is selected from a water extract and / or an alcohol extract.

[0061] In some embodiments, the composition further comprises 2'-methoxymatrine and / or pyralidinol A.

[0062] In some embodiments, the added amount of 2'-methoxymatrine in the composition is 0.1~1μM, including but not limited to 0.1μM, 0.15μM, 0.2μM, 0.25μM, 0.3μM, 0.35μM, 0.4μM, 0.45μM, 0.5μM, 0.55μM, 0.6μM, 0.65μM, 0.7μM, 0.75μM, 0.8μM, 0.85μM, 0.9μM, 0.95μM, 1μM or any range formed by any two of the foregoing and any numerical value within the range.

[0063] In some embodiments, the added amount of Panlongol A in the composition is 0.05~0.5μM, including but not limited to 0.05μM, 0.08μM, 0.1μM, 0.12μM, 0.15μM, 0.18μM, 0.2μM, 0.22μM, 0.25μM, 0.28μM, 0.3μM, 0.32μM, 0.35μM, 0.38μM, 0.4μM, 0.42μM, 0.45μM, 0.48μM, 0.5μM or any range formed by any two of the foregoing and any numerical value within the range.

[0064] In some embodiments, the amount of mangosteen added to the composition is 1-5 μg / mL, including but not limited to 1 μg / mL, 1.2 μg / mL, 1.5 μg / mL, 1.8 μg / mL, 2 μg / mL, 2.2 μg / mL, 2.5 μg / mL, 2.8 μg / mL, 3 μg / mL, 3.2 μg / mL, 3.5 μg / mL, 3.8 μg / mL, 4 μg / mL, 4.2 μg / mL, 4.5 μg / mL, 4.8 μg / mL, 5 μg / mL or a range formed by any two of the foregoing and any value within the range.

[0065] In some embodiments, the amount of Cornus officinalis added to the composition is 10-20 μg / mL, including but not limited to 10 μg / mL, 11 μg / mL, 12 μg / mL, 13 μg / mL, 14 μg / mL, 14.2 μg / mL, 15 μg / mL, 16 μg / mL, 17 μg / mL, 18 μg / mL, 18.2 μg / mL, 19 μg / mL, 20 μg / mL or a range formed by any two of the foregoing and any value within the range.

[0066] In some embodiments, the composition comprises 10-20 μg / mL Cornus officinalis extract, 1-5 μg / mL Mangosteen extract and 0.1-1 μM 2'-methoxymatrine.

[0067] In some embodiments, the composition comprises 10-20 μg / mL Cornus officinalis extract, 1-5 μg / mL Mangosteen extract and 0.05-0.5 μM Penicillin A.

[0068] In some embodiments, the composition comprises 10-20 μg / mL Cornus officinalis extract, 1-5 μg / mL Mangosteen extract, 0.1-1 μM 2'-methoxymatrine and 0.05-0.5 μM Penicillin A.

[0069] The compositions and proportions of the above compositions can achieve positive effects in treating anemia. Furthermore, the composition comprising 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine and 0.1 μM Penrophyllum A can better achieve the therapeutic effect.

[0070] In some embodiments, the iron supplement preparation achieves the iron supplement effect by promoting intestinal iron absorption and / or inhibiting hepcidin.

[0071] In a second aspect, the present invention also provides a complex, the components of which include Cornus officinalis and Mangosteen, the Cornus officinalis includes Cornus officinalis medicinal materials and / or Cornus officinalis extracts, the mangosteen includes Mangosteen medicinal materials and / or Mangosteen extracts, and also contains 2'-methoxymatrine and / or panlongol A.

[0072] In some embodiments, the cornus officinalis can be the flower of cornus officinalis, the bark, root, stem and other parts of cornus officinalis and their extracts, and the cornus officinalis extract includes alkaloids, polysaccharides, saponins, flavonoids and polyphenols;

[0073] In some embodiments, the Cornus officinalis extract is selected from a water extract and / or an alcohol extract.

[0074] In some embodiments, the mangosteen and / or mangosteen extract can be mangosteen pulp, mangosteen peel, mangosteen flower, mangosteen bark root, stem and other parts and extracts thereof, and the mangosteen extract includes alkaloids, polysaccharides, saponins, flavonoids and polyphenols and other substances;

[0075] In some embodiments, the mangosteen extract is selected from a water extract and / or an alcohol extract.

[0076] In some embodiments, the content ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM, including but not limited to 10:1:0.1, 10:2:0.3, 10:3:0.4, 10:4:0.5, 10:5:1, 12:1:0.1, 14:1:0.3, 16:1:0.4, 18:1:0.5, 18.2:1:0.1, 18.2:1:0.3, 20:1:0.1, 12:2:0.1, 14:2:0.3, 16:2:0. 4, 18:2:0.5, 20:2:0.1, 12:3:0.1, 14:3:0.2, 15:3:0.3, 16:3.2:0.3, 18:3:0.4, 20:3:1, 12:4:0.1, 14:4:0.2, 15:4:0.3, 16:4:0.3, 18:4:0.5, 20:4:1, 12:5:0.1, 14:5:0.2, 14.2:5:0.1, 14.2:5:0.3, 16:5:0.3, 18:5:0.5, 20:5:1, or any ratio within the range formed by any two of the foregoing.

[0077] In some embodiments, the content ratio of Cornus officinalis, Mangosteen and Panlong phenol A in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.05-0.5) μM, including but not limited to 10:1:0.05, 10:2:0.1, 10:3:0.2, 10:4:0.3, 10:5:0.5, 12:1:0.05, 14:1:0.1, 16:1:0.2, 18:1:0.3, 18.2:1:0.1, 18.2:1:0.3, 20:1:0.5, 12:2:0.05, 14:2:0.1, 16:2:0.2, 0.3, 20:2:0.5, 12:3:0.05, 14:3:0.1, 15:3:0.2, 16:3.2:0.3, 18:3:0.3, 20:3:0.5, 12:4:0.1, 14:4:0.2, 15:4:0.3, 16:4:0.3, 18:4:0.3, 20:4:0.5, 12:5:0.05, 14:5:0.1, 14.2:5:0.1, 14.2:5:0.3, 16:5:0.2, 18:5:0.3, 20:5:0.5 or any ratio within the range formed by any two of the foregoing.

[0078] In some embodiments, the mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlongol A in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM: (0.05-0.5) μM, including but not limited to 10:1:0.1:0.05, 10:2:0.3:0.1, 10:3:0.3:0.2, 10:4:0.5:0.3, 10:5:1:0.5, 12:1 :0.1:0.05, 14:1:0.3:0.1, 16:1:0.3:0.3, 18:1:0.5:0.3, 18.2:1:0.3:0.1, 18.2:1:0.1:0.3, 20:1:1:0.5, 12:2:0.1:0.05, 14:2:0.3:0.1, 16:2:0.3:0.3, 18:2:0.5:0.3 、20:2:1:0.5、12:3:0.1:0.05、14:3:0.3:0.1、15:3:0.3:0.3、16:3.2:0.5:0.3、18:3:1:0.5、20:3:0.1:0.5、12:4:0.2:0.05、14:4:0.3:0.1、15:4:0.3:0.2、16:4:0.3:0 .3, 18:4:0.5:0.4, 20:4:1:0.5, 12:5:0.1:0.05, 14:5:0.3:0.1, 14.2:5:0.3:0.1, 14.2:5:0.1:0.3, 16:5:0.3:0.3, 18:5:0.5:0.3, 20:5:1:0.5 or any ratio within the range formed by any two of the foregoing.

[0079] In some embodiments, the mass ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the composite is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM.

[0080] In some embodiments, the mass ratio of Cornus officinalis, Mangosteen and Panlongphenol A in the complex is (15~18) μg / mL: (3~4) μg / mL: (0.05~0.2) μM.

[0081] In some embodiments, the mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlongol A in the complex is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM: (0.05-0.2) μM.

[0082] In some embodiments, the composition of the complex includes 10-20 μg / mL cornus extract, 1-5 μg / mL mangosteen extract, 0.1-1 μM 2'-methoxymatrine and 0.05-0.5 μM panlongol A, and the complex has good compatibility and has a good effect in treating anemia. Further, the composition of the complex includes 16 μg / mL cornus extract, 3.2 μg / mL mangosteen extract, 0.3 μM 2'-methoxymatrine and 0.1 μM panlongol A, which can better achieve the above effect.

[0083] In a third aspect, the present invention also provides a drug for treating anemia, wherein the drug comprises the complex and pharmaceutically acceptable excipients.

[0084] In some embodiments, the pharmaceutically acceptable excipients include natural excipients, semi-synthetic and / or synthetic excipients, such as one or more of solvents, propellants, solubilizers, suspending agents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, glidants, compression aids, flavoring agents, preservatives, suspending agents, coating agents and fragrances.

[0085] In some embodiments, the drug is in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes or aerosols.

[0086] In some embodiments, the dosage form of the drug includes oral dosage, injection, patch, liniment, spray, inhalation, drops, lotion or suppository.

[0087] In some embodiments, the anemia is iron deficiency anemia, autoimmune hemolytic anemia or nausea anemia. Furthermore, the complex has the activity of reversing functional iron deficiency, thereby improving the body's iron deficiency abnormality. Therefore, the complex has a better therapeutic effect on iron deficiency anemia.

[0088] In some embodiments, the medicament is for treating anemia by promoting intestinal iron absorption and / or inhibiting hepcidin.

[0089] In a fourth aspect, the present invention further provides a health product for replenishing qi and blood, the ingredients of the health product comprising the complex and auxiliary materials.

[0090] In some embodiments, the excipients include natural excipients, semi-synthetic and / or synthetic excipients, such as one or more of solvents, propellants, solubilizers, suspending agents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, glidants, compression aids, flavoring agents, preservatives, suspending agents, coating agents and fragrances.

[0091] In some embodiments, the health care product is in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes or aerosols.

[0092] In some embodiments, the dosage form of the health care product includes oral preparations, injections, patches, ointments, sprays, inhalants, drops, lotions or suppositories.

[0093] In some embodiments, the health care product can promote intestinal iron absorption and / or inhibit hepcidin, thereby achieving the effects of improving iron deficiency anemia, replenishing blood and invigorating Qi, etc.

[0094] In a fifth aspect, the present invention also provides a multifunctional food for improving sub-health status, wherein the ingredients of the food include the complex and food additives.

[0095] The food additives include, but are not limited to, acidity regulators, anticaking agents, defoaming agents, antioxidants, bleaching agents, leavening agents, colorants, color preservatives, enzyme preparations, flavor enhancers, nutritional fortifiers, preservatives, sweeteners, thickeners or spices, etc.

[0096] For experimental parameters not specified in the following specific embodiments, reference is made to the instructions given in the present application document, and reference may also be made to experimental manuals in the art or other experimental methods known in the art, or to experimental conditions recommended by manufacturers.

[0097] The raw materials and reagents involved in the following specific examples can be obtained from commercial sources, or can be prepared by those skilled in the art according to known methods.

[0098] Among them: human colon adenocarcinoma Caco-2 cells were purchased from Xiamen Yimo Biotechnology Co., Ltd., China; human liver cancer cell HepG2 cells were purchased from Saiye Biotechnology Co., Ltd., China; DMEM culture medium and FBS fetal bovine serum were purchased from Gibco, USA; Cell Counting Kit (CCK-8) cell proliferation-toxicity detection kit was purchased from Tongren Chemical Research Institute; Taq Pro Universal SYBR qPCR Master Mix was purchased from Nanjing Novozyme Biotechnology Co., Ltd., China; RNA-easy Isolation Reagent was purchased from Nanjing Novazon Biotech Co., Ltd., China; FastKing one-step genomic cDNA first-strand synthesis premix was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., China; FerroOrange kit, a divalent iron ion detection probe, was purchased from Dojindo Chemical Research Institute, Japan; 2'-methoxymatrine (CAS: 270249-38-2) and synthetic panlongol A (CAS: 126192-35-6) were purchased from Tianjin WuXi AppTec; Cornus officinalis extract (Cornus officinalis fruit pulp water-soluble extract, 10:1) and mangosteen extract (mangosteen peel extract 10:1) were purchased from Fufeng Snot Biotechnology Co., Ltd.

[0099] Example 1: Application of the composition in the preparation of iron supplement preparations

[0100] The composition formula provided in this embodiment is as follows:

[0101] Formula 1: 18.2μg / mL Cornus officinalis extract and 1μg / mL Mangosteen extract.

[0102] Formula 2: 16μg / mL Cornus officinalis extract and 3.2μg / mL Mangosteen extract.

[0103] Formula 3: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract and 0.3 μM 2'-methoxymatrine.

[0104] Formula 4: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract and 0.1 μM Panax notoginseng A.

[0105] Formula 5: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine and 0.1 μM Panax notoginseng A.

[0106] Formula 6: 14.2 μg / mL Cornus officinalis extract, 5 μg / mL Mangosteen extract, 0.1 μM 2'-methoxymatrine and 0.3 μM Panax notoginseng A.

[0107] The compositions of the above six groups of formulas all have the effect of supplementing iron.

[0108] Test Example 1: Cytotoxicity test of the composition

[0109] In this test example, the CCK-8 method was used to screen out the concentration of the composition that had no significant inhibitory effect on the viability of Caco-2 cells and HepG2 cells. The specific method was as follows:

[0110] Caco-2 cells and HepG2 cells in the logarithmic growth phase were taken, digested and made into cell suspension with complete medium, and inoculated into 96-well plates, with 100 μL of cell suspension inoculated into each well and the cell number was 1.2×10 4 / well, cultured overnight in a constant temperature incubator at 37°C and 5% CO2, and when the cell confluence was about 80%, the test substances were added respectively: the six groups of formula compositions provided in Example 1, 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine, and 0.1 μM Panlongol A to treat the two types of cells.

[0111] The blank group was set as the medium and cells without the test substance, the control group was set as the medium and cells without the test substance, and the experimental group was set as the medium and cells containing the test substance. Then each group was placed in a 37°C, 5% CO2 constant temperature incubator for 24 hours. After the original culture medium was replaced with a culture medium containing 10% CCK-8, the 96-well plate was placed in a constant temperature incubator for 4 hours, and the OD value of each well was measured at a wavelength of 450nm. The proliferation and survival rate of cells by different test substances was calculated according to formula (I), and 3 replicates were performed for each concentration.

[0112] (I)

[0113] The results of the effects of each group on the survival rate of Caco-2 cells and HepG2 cells obtained by the above test method are shown in Table 1, Figure 1 and Figure 2:

[0114] Table 1: Effects of the test substances on the survival rate of Caco-2 cells and HepG2 cells ( ±s, n=3)

[0115]

[0116] The results in Table 1 and Figures 1 and 2 show that the extract of Cornus officinalis, the extract of Mangosteen, 2'-methoxymatrine, Panlongol A and the six groups of combination formulas had no significant inhibitory effect on the survival rate of Caco-2 cells and HepG2 cells.

[0117] Experimental Example 2: Fe uptake by Caco-2 cells of the composition 2+ Impact

[0118] This test method uses FerroOrange fluorescent probe to detect the effect of the composition on promoting the absorption of iron ions Fe by Caco-2 cells. 2+ The specific test method is as follows:

[0119] Caco-2 cells in the logarithmic growth phase were taken, digested and made into a cell suspension with complete medium, and inoculated into a 96-well plate, with 100 μL of cell suspension inoculated into each well and the cell number was 1.2×10 4 / well, cultured in a constant temperature incubator at 37°C and 5% CO2, and when the cell confluence was about 80%, 16μg / mL Cornus officinalis extract, 3.2μg / mL Mangosteen extract, 0.3μM 2'-methoxymatrine, 0.1μM Panlong phenol A and the six groups of composition formulas provided in Example 1 were added to treat Caco-2 cells respectively. After 12h of treatment, the culture medium was discarded, and 100μL of complete culture medium containing 4μg / mL ferric citrate and 10μM ferrous sulfate was added to each well. After overnight culture, the culture medium was discarded, washed 3 times with Hank's salt solution, 1μM FerroOrange working solution was added and placed in a constant temperature incubator at 7°C and 5% CO2 for 30min. At the end of incubation, Hoechst 33342 dye was added and incubated for 5min. The fluorescence value was detected on the machine, and the results are shown in Table 2 and Figure 3.

[0120] Table 2: Effects of the test substances on Fe uptake by Caco-2 cells 2+ The impact of

[0121]

[0122] Note: Compared with a single substance, * represents p﹤0.05, ** represents p﹤0.01;

[0123] The results in Table 2 and Figure 3 show that compared with a single substance, the six groups of compositions enhanced the ability of Caco-2 cells to take up iron ions, which was statistically significant compared with a single substance.

[0124] Experimental Example 3: Effect of the Composition on the mRNA Expression of Hepcidin in HepG2 Cells

[0125] In this experiment, 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine, 0.1 μM Panlongphenol A and six groups of composition formulas provided in Example 1 were selected to treat HepG2 cells respectively, and RNA was extracted and reverse transcribed and qPCR experiments were performed to evaluate the mRNA expression level of Hepcidin gene in HepG2 cells. The specific method is as follows:

[0126] HepG2 cells in the logarithmic growth phase were taken and digested, and then prepared into a cell suspension using DMEM complete medium containing 15% fetal bovine serum and 1% double antibody. 1×10 6 In a 6 cm cell culture dish, 5 mL per well, cultured in a constant temperature incubator at 37 ° C and 5% CO2. After 72 hours of adaptive culture, the control group was replaced with fresh complete medium, and the intervention group was replaced with complete medium containing the corresponding monomers and compositions. After 12 hours of culture in a constant temperature incubator at 37 ° C and 5% CO2, the RNA of each group of cells was extracted according to the RNA-easy Isolation Reagent instructions, and the RNA concentration was detected. According to the instructions of the FastKing one-step genomic cDNA first-chain synthesis premix reagent, the extracted RNA of each group was reverse transcribed into cDNA at a concentration of 100 ng / μL. Finally, the experimental system was prepared according to the instructions of the TaqPro Universal SYBR qPCR Master Mix to detect the mRNA expression level of the Hepcidin gene. The results are shown in Table 3 and Figure 4.

[0127] Table 3: Effects of the test substances on the mRNA expression of Hepcidin gene

[0128]

[0129] Note: Compared with a single substance, * represents p﹤0.05;

[0130] The results in Table 3 and Figure 4 show that, compared with a single substance, the six groups of compositions can significantly reduce the mRNA expression of Hepcidin, which is statistically significant compared with a single substance.

[0131] Experimental Example 4: Effect of the Composition on the Protein Expression of Hepcidin in HepG2 Cells

[0132] In this experiment, 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine, 0.1 μM Panlongphenol A and the six groups of compositions provided in Example 1 were selected to treat HepG2 cells respectively, and Western blot experiments were performed after protein extraction to evaluate the expression level of Hepcidin protein in HepG2 cells. The specific method is as follows:

[0133] HepG2 cells in the logarithmic growth phase were digested and made into a cell suspension using DMEM complete medium containing 10% fetal bovine serum and 1% double antibody. 1×10 6In a 6 cm cell culture dish, 5 mL per well, cultured in a 37°C, 5% CO2 constant temperature incubator. After 72 hours of adaptive culture, the control group was replaced with fresh complete culture medium, and the intervention group was replaced with complete culture medium containing the above monomers and mixtures. After 12 hours of culture in a 37°C, 5% CO2 constant temperature incubator, the cells were scraped with a cell spatula, centrifuged at 1000 rpm for 5 min, lysed on ice to extract total cell protein, and the protein concentration was determined by BCA kit; 30 μg of protein was taken from each group for SDS-PAGE electrophoresis, transferred to PVDF membrane, blocked with 5% BSA, added with Hepcidin and GAPDH primary antibodies, placed in a 4°C shaker overnight incubation, washed with TBS-T washing solution, added with HRP-labeled secondary antibody and incubated at room temperature for 1 hour, washed with TBS-T washing solution, and photographed with a chemiluminescence imaging system. The data results are shown in Tables 4 and Figure 5 shown.

[0134] Table 4: Effects of the test substances on the protein expression of Hepcidin

[0135]

[0136] Note: Compared with a single substance, * represents p﹤0.05;

[0137] From Table 4 and Figure 5 The results showed that compared with a single substance, the six groups of combinations could significantly reduce the protein expression of Hepcidin, which was statistically significant compared with a single substance.

[0138] GraphPad Prism 9.3.0 statistical software was used for data processing, and some experimental results were expressed as Mean±SD.

[0139] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0140] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Use of the composition in the preparation of an iron supplement preparation, characterized in that: The composition comprises cornus officinalis and mangosteen, wherein the cornus officinalis comprises cornus officinalis medicinal materials and / or cornus officinalis extract, and the mangosteen comprises mangosteen medicinal materials and / or mangosteen extract.

2. The use according to claim 1, characterized in that: The composition satisfies at least one of the following (1) to (3): (1) The content ratio of Cornus officinalis to Mangosteen in the composition is (10-20): (1-5); (2) The Cornus officinalis extract is selected from a water extract and / or an alcohol extract; (3) The mangosteen extract is selected from water extract and / or alcohol extract.

3. The use according to claim 1 or 2, characterized in that: The composition also includes 2'-methoxymatrine and / or panlongol A.

4. The use according to claim 3, characterized in that: The composition satisfies the following (1) and / or (2): (1) The amount of 2'-methoxymatrine added to the composition is 0.1-1 μM; (2) The amount of Panlongol A added to the composition is 0.05~0.5 μM.

5. The composition according to claim 4, characterized in that The composition comprises 10-20 μg / mL cornus officinalis extract, 1-5 μg / mL mangosteen extract, 0.1-1 μM 2'-methoxymatrine and 0.05-0.5 μM pyrocalyx A.

6. The use according to claim 1 or 2, characterized in that: The iron supplement preparation achieves the iron supplement effect by promoting intestinal iron absorption and / or inhibiting hepcidin.

7. A composite, characterized in that The components of the complex include cornus officinalis and mangosteen, the cornus officinalis includes cornus officinalis medicinal materials and / or cornus officinalis extract, the mangosteen includes mangosteen medicinal materials and / or mangosteen extract, and also contains 2'-methoxymatrine and / or panlongol A.

8. The composite according to claim 7, characterized in that The composite satisfies at least one of the following (1) to (3): (1) The content ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM; (2) The mass ratio of Cornus officinalis, Mangosteen and Panlongphenol A in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.05-0.5) μM; (3) The mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlongol A in the composite is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM: (0.05-0.5) μM.

9. The composite according to claim 7 or 8, characterized in that The composite satisfies at least one of the following (1) to (5): (1) The mass ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the composite is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM; (2) The mass ratio of Cornus officinalis, Mangosteen and Panlongphenol A in the composite is (15-18) μg / mL: (3-4) μg / mL: (0.05-0.2) μM; (3) The mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlongol A in the composite is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM: (0.05-0.2) μM; (4) The Cornus officinalis extract is selected from a water extract and / or an alcohol extract; (5) The mangosteen extract is selected from water extract and / or alcohol extract.

10. The composite according to claim 9, characterized in that The composition of the complex includes 10-20 μg / mL cornus officinalis extract, 1-5 μg / mL mangosteen extract, 0.1-1 μM 2'-methoxymatrine and 0.05-0.5 μM panlongol A.

11. A drug for treating anemia, characterized in that: The composition of the drug comprises the complex according to any one of claims 7 to 10, and pharmaceutically acceptable excipients.

12. The drug for treating anemia according to claim 11, characterized in that: The drug satisfies at least one of the following (1) to (4): (1) The drug is in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes or aerosols; (2) The dosage form of the drug includes oral preparation, injection, patch, ointment, spray, inhalation, drops, lotion or suppository; (3) The anemia is iron deficiency anemia, autoimmune hemolytic anemia or nausea anemia; (4) The drug is used to treat anemia by promoting intestinal iron absorption and / or inhibiting hepcidin.

13. A health product for replenishing qi and blood, characterized in that: The ingredients of the health care product include the compound according to any one of claims 7 to 10 and auxiliary materials.

14. The health product for replenishing qi and blood according to claim 13, characterized in that: The health care product is in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes or aerosols.

15. A multifunctional food for improving sub-health status, characterized in that: The ingredients of the food include the compound according to any one of claims 7 to 10 and a food additive.

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