Use of composition in preparing iron-supplementing formulation

Abstract

Use of a composition in preparing an iron-supplementing formulation. The composition comprises Cornus officinalis and / or an extract thereof and Garcinia mangostana and / or an extract thereof, and may further comprise 2'-methoxykurarinone and / or spiranthol A. The composition can promote intestinal iron absorption and / or inhibit hepcidin, treat functional iron deficiency, regulate the constitution, tonify deficiency and replenish qi, and regulate physical health state.

Description

Application of the composition in the preparation of iron supplements

[0001] Related applications

[0002] This application claims priority to Chinese patent application filed on January 22, 2025, with application number 202510098630.5, entitled "Use of a composition in the preparation of an iron supplement", the entire contents of which are incorporated herein by reference. Technical Field

[0003] This application relates to the field of traditional Chinese medicine preparation technology, and in particular to the application of the composition in the preparation of iron supplements. Background Technology

[0004] From single-celled bacteria to multicellular organisms (such as humans), iron is one of the most essential metals for maintaining life. Iron plays a crucial role in several cellular processes, including DNA synthesis, nucleic acid repair, mitochondrial cellular respiration, cell growth, and cell death, and also contributes to host defense and cell signaling. Most of the iron in the human body is related to hemoglobin in red blood cells; iron deficiency leads to anemia, which in turn causes a range of health problems, such as poor blood circulation and cold hands and feet.

[0005] Iron absorbed by intestinal cells can be directly used in intracellular metabolic mechanisms, stored in ferritin, or released through the basement membrane for systemic iron delivery. Most iron absorption in the body comes from food. 3+ It needs to be reduced to Fe first. 2+ Iron is absorbed by the intestines and then transported to the liver for storage. Hepcidin, released by the liver, is a key regulator of iron homeostasis. It binds to the only known iron export protein, femoral transfer protein (FPN), mediating FPN ubiquitination, internalization, and degradation, and directly blocking this channel, thus preventing iron from being exported from cells to plasma. When serum iron levels are elevated, hepcidin expression is upregulated, leading to impaired iron transport through FPN to plasma, thus providing a negative feedback loop to prevent the accumulation of potentially toxic iron in the body. Decreased plasma iron levels lead to reduced transferrin saturation, resulting in decreased hepcidin synthesis. Therefore, hepcidin expression is enhanced, downregulating the expression of the iron transport protein (ferroportin), reducing the body's absorption of iron. When the body is iron deficient, the above processes are reversed, thereby maintaining iron homeostasis.

[0006] Therefore, in order to improve the series of adverse effects caused by iron deficiency, it has become more important to explore safe and effective iron-supplementing bioactive substances. Summary of the Invention

[0007] Based on this, in a first aspect, this application provides the use of a composition in the preparation of an iron supplement, wherein the composition comprises Cornus officinalis and mangosteen, wherein the Cornus officinalis comprises at least one of Cornus officinalis medicinal material and Cornus officinalis extract, and the mangosteen comprises at least one of mangosteen medicinal material and mangosteen extract.

[0008] In some embodiments, the composition satisfies at least one of (1) to (3) below:

[0009] (1) The mass ratio of Cornus officinalis and Mangosteen in the composition is (10-20):(1-5);

[0010] (2) The Cornus officinalis extract is selected from at least one of water extract and alcohol extract;

[0011] (3) The mangosteen extract is selected from at least one of water extract and alcohol extract.

[0012] In some embodiments, the composition further includes at least one of 2'-methoxymatrine and Panlongphenol A.

[0013] In some embodiments, the composition satisfies at least one of (1) and (2) below:

[0014] (1) The amount of 2'-methoxymatrine added in the composition is 0.1-1 μM;

[0015] (2) The amount of Panlong phenol A added in the composition is 0.05-0.5 μM.

[0016] 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 Panlongphenol A.

[0017] In some embodiments, the iron supplement achieves its iron-supplementing effect by promoting intestinal iron absorption.

[0018] In some embodiments, the iron supplementation agent achieves its iron-supplementing effect by inhibiting hepcidin.

[0019] Secondly, this application also provides a complex comprising Cornus officinalis and mangosteen, wherein the Cornus officinalis comprises at least one of Cornus officinalis medicinal material and Cornus officinalis extract, and the mangosteen comprises at least one of mangosteen medicinal material and mangosteen extract, and further comprises at least one of 2'-methoxymatrine and Panlongphenol A.

[0020] In some embodiments, the complex satisfies at least one of (1) to (3) below:

[0021] (1) The content ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the complex is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM;

[0022] (2) The mass ratio of Cornus officinalis, Mangosteen and Panlong phenolic A in the complex is (10-20) μg / mL: (1-5) μg / mL: (0.05-0.5) μM;

[0023] (3) The mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlong phenol A in the complex is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM: (0.05-0.5) μM.

[0024] In some embodiments, the complex satisfies at least one of the following (1) to (5):

[0025] (1) The mass ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the complex is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM;

[0026] (2) The mass ratio of Cornus officinalis, Mangosteen and Panlong phenolic A in the complex is (15-18) μg / mL: (3-4) μg / mL: (0.05-0.2) μM;

[0027] (3) The mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlong phenolic A in the complex is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM: (0.05-0.2) μM;

[0028] (4) The Cornus officinalis extract is selected from at least one of water extract and alcohol extract;

[0029] (5) The mangosteen extract is selected from at least one of water extract and alcohol extract.

[0030] In some embodiments, the complex 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 Panlongphenol A.

[0031] Thirdly, this application also provides a medicament for treating anemia, the composition of which includes the complex provided in the second aspect, and pharmaceutically acceptable excipients.

[0032] In some embodiments, the drug satisfies at least one of the following (1) to (5):

[0033] (1) The form of the drug includes tablets, granules, capsules, solutions, powders, suspensions, emulsions, ointments or aerosols;

[0034] (2) The dosage forms of the drug include oral preparations, injections, patches, liniments, sprays, inhalers, drops, lotions, or suppositories;

[0035] (3) The anemia mentioned is iron deficiency anemia, autoimmune hemolytic anemia, or nausea anemia;

[0036] (4) The drug treats anemia by promoting intestinal iron absorption;

[0037] (5) The drug treats anemia by inhibiting hepcidin.

[0038] Fourthly, this application also provides a health product for invigorating qi and nourishing blood, the ingredients of which include the complex and excipients provided in the second aspect.

[0039] In some embodiments, the health product may be in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes, or aerosols.

[0040] Fifthly, this application also provides a multifunctional food for improving sub-health conditions, the ingredients of which include the complex and food additives provided in the second aspect. Attached Figure Description

[0041] To more clearly illustrate the technical solutions in the embodiments of this application or the conventional technology, the drawings used in the description of the embodiments or the conventional technology will be briefly introduced below. Obviously, the drawings described below are only embodiments of this application. For those skilled in the art, other drawings can be obtained based on the disclosed drawings without creative effort.

[0042] The accompanying drawings, which form part of this application, are used to provide a further understanding of this application. The illustrative embodiments and descriptions of this application are used to explain this application and do not constitute an undue limitation of this application. In the drawings:

[0043] Figure 1 shows the effects of each individual substance and formulations 1 to 6 of the combination on the survival rate of Caco-2 cells in Experiment Example 1.

[0044] Figure 2 shows the effects of each individual substance and formulations one through six in Experiment 1 on the survival rate of HepG2 cells.

[0045] Figure 3 shows the statistical data on the absorption of iron ions by Caco-2 cells by each single substance and the combination formulations one to six in Experimental Example 2.

[0046] Figure 4 shows the statistical data on the mRNA expression of the Hepcidin gene in HepG2 cells by each single substance and the combination formulations one to six in Experiment Example 3.

[0047] Figure 5 shows the statistical data on the expression level of Hepcidin protein in HepG2 cells by each single substance and the combination formulations one to six in Experiment Example 4. Detailed Implementation

[0048] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0049] To facilitate understanding of this application, preferred embodiments of this application are provided below for a more comprehensive description of the technical solutions of this application. However, this application can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of the disclosure of this application. It should be noted that experimental methods in the following embodiments of this application that do not specify specific conditions are generally performed under conventional conditions or according to the conditions recommended by the manufacturer. All commonly used chemical reagents used in the embodiments are commercially available products.

[0050] It should be noted that experimental methods in the following embodiments of this application, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. All commonly used chemical reagents used in the embodiments are commercially available products.

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0052] Cornus officinalis (scientific name: *Cornus officinalis* Sieb. et Zucc.), also known as Shan Yu Rou, Shan Yu Rou, or Shan Yu Rou, is the dried, ripe pulp of the fruit of *Cornus officinalis* Sieb. et Zucc., a plant in the Cornaceae family. It has a sour and astringent taste, is slightly warm in nature, and enters the liver and kidney meridians. It has the effects of tonifying the liver and kidneys, and astringing and consolidating the body.

[0053] Mangosteen (scientific name: *Garcinia mangostana* L.), also known as mangosteen fruit, is neutral in nature and has a sweet and slightly sour taste. Mangosteen peel is cool in nature and bitter and astringent in taste; it strengthens the spleen, promotes the production of body fluids, and stops diarrhea. Mangosteen peel also has anti-inflammatory and analgesic properties, and is used to treat spleen deficiency diarrhea, dry mouth, burns, scalds, eczema, and stomatitis.

[0054] In a first aspect, this application provides the use of a composition in the preparation of an iron supplement, wherein the composition comprises Cornus officinalis and mangosteen, wherein the Cornus officinalis comprises at least one of Cornus officinalis medicinal material and Cornus officinalis extract, and the mangosteen comprises at least one of mangosteen medicinal material and mangosteen extract.

[0055] This application, through the compatibility of the above-mentioned ingredients, has discovered that multiple components in each formula have a synergistic effect in promoting iron absorption, thus playing a role in supplementing the body with iron.

[0056] In some embodiments, the mass 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 that range.

[0057] In some embodiments, the Cornus officinalis medicinal material includes the pulp, pericarp, flowers, and bark of the Cornus officinalis tree.

[0058] In some embodiments, the Cornus officinalis extract includes substances extracted from the pulp, pericarp, flowers, and bark of Cornus officinalis.

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

[0060] In some embodiments, the Cornus officinalis extract is selected from at least one of an aqueous extract and an alcoholic extract.

[0061] In some embodiments, the mangosteen medicinal material includes the pulp, peel, flowers, bark, roots, and stems of the mangosteen.

[0062] In some embodiments, the mangosteen extract includes substances extracted from the pulp, pericarp, flowers, bark, roots, stems, and other parts of the mangosteen.

[0063] Optionally, mangosteen extract includes substances such as alkaloids, polysaccharides, saponins, flavonoids, and polyphenols.

[0064] In some embodiments, the mangosteen extract is selected from at least one of an aqueous extract and an alcoholic extract.

[0065] In some embodiments, the composition further includes at least one of 2'-methoxymatrine and Panlongphenol A.

[0066] In some embodiments, the amount of 2'-methoxymatrine added to the composition is 0.1 to 1 μM, including but not limited to 0.1 μM, 0.15 μM, 0.2 μM, 0.25 μM, 0.3 μM, 0.35 μM, 0.4 μM, 0.45 μM, 0.5 μM, 0.55 μM, 0.6 μM, 0.65 μM, 0.7 μM, 0.75 μM, 0.8 μM, 0.85 μM, 0.9 μM, 0.95 μM, 1 μM, or any range formed by any two of the foregoing and any value within the range.

[0067] In some embodiments, the amount of Panlongphenol A added to the composition is 0.05 to 0.5 μM, including but not limited to 0.05 μM, 0.08 μM, 0.1 μM, 0.12 μM, 0.15 μM, 0.18 μM, 0.2 μM, 0.22 μM, 0.25 μM, 0.28 μM, 0.3 μM, 0.32 μM, 0.35 μM, 0.38 μM, 0.4 μM, 0.42 μM, 0.45 μM, 0.48 μM, 0.5 μM, or any range formed by any two of the foregoing and any value within the range.

[0068] In some embodiments, the amount of mangosteen added to the composition is 1 to 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 any range formed by any two of the foregoing and any value within the range.

[0069] In some embodiments, the amount of Cornus officinalis added to the composition is 10 to 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 any range formed by any two of the foregoing and any value within the range.

[0070] 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.

[0071] 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 Panlongphenol A.

[0072] 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 Panlongphenol A.

[0073] Within the composition and proportion range of the above-mentioned compositions, positive effects can be achieved in treating anemia. Furthermore, the composition of the above-mentioned compositions includes 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine and 0.1 μM Panlongphenol A, which can better achieve this therapeutic effect.

[0074] In some embodiments, the iron supplement achieves its iron-supplementing effect by promoting intestinal iron absorption.

[0075] In some embodiments, the iron supplementation agent achieves its iron-supplementing effect by inhibiting hepcidin.

[0076] Secondly, this application also provides a complex comprising Cornus officinalis and mangosteen, wherein the Cornus officinalis comprises at least one of Cornus officinalis medicinal material and Cornus officinalis extract, and the mangosteen comprises at least one of mangosteen medicinal material and mangosteen extract, and further comprises at least one of 2'-methoxymatrine and Panlongphenol A.

[0077] In some embodiments, the Cornus officinalis can be the flowers, bark, roots, stems, and other parts of Cornus officinalis and their extracts, and the Cornus officinalis extract includes substances such as alkaloids, polysaccharides, saponins, flavonoids, and polyphenols;

[0078] In some embodiments, the Cornus officinalis extract is selected from at least one of an aqueous extract and an alcoholic extract.

[0079] In some embodiments, the mangosteen and mangosteen extract may be the pulp, pericarp, flower, bark, root, stem, and other parts of the mangosteen and their extracts. The mangosteen extract includes substances such as alkaloids, polysaccharides, saponins, flavonoids, and polyphenols.

[0080] In some embodiments, the mangosteen extract is selected from at least one of an aqueous extract and an alcoholic extract.

[0081] In some embodiments, the content ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the complex 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, and 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 of the above two ranges and any ratio within the range.

[0082] In some embodiments, the content ratio of Cornus officinalis, Mangosteen, and Panlongphenol A in the complex 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, 18:2:0.3, 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 of the ranges formed by the foregoing and any ratio within the range.

[0083] In some embodiments, the mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine, and Panax notoginseng A in the complex 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, and 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, 1 8:2:0.5:0.3, 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 range formed by any two of the foregoing and any ratio within that range.

[0084] In some embodiments, the mass ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the complex is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM.

[0085] 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.

[0086] In some embodiments, the mass ratio of Cornus officinalis, Mangosteen, 2'-methoxymatrine and Panlong phenolic A in the complex is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM: (0.05-0.2) μM.

[0087] In some embodiments, the complex 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 Pantothecin A. This complex exhibits good compatibility and is effective in treating anemia. Furthermore, the complex comprising 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine, and 0.1 μM Pantothecin A further enhances the aforementioned effects.

[0088] Thirdly, this application also provides a medicament for treating anemia, the composition of which includes the complex provided in the second aspect, and pharmaceutically acceptable excipients.

[0089] In some embodiments, the pharmaceutically acceptable excipient comprises at least one of natural, semi-synthetic, and synthetic excipients, such as solvents, propellants, solubilizers, suspending agents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, flow aids, pressure aids, flavoring agents, preservatives, suspending agents, coating agents, and flavoring agents.

[0090] In some embodiments, the form of the drug includes tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes, or aerosols.

[0091] In some embodiments, the dosage form of the drug includes oral dosage, injection, patch, liniment, spray, inhaler, drops, lotion, or suppository.

[0092] In some embodiments, the anemia is iron deficiency anemia, autoimmune hemolytic anemia, or nausea anemia. Furthermore, the complex has reversible functional iron deficiency activity, thereby improving the body's iron deficiency abnormalities; therefore, the complex is particularly effective for iron deficiency anemia.

[0093] In some embodiments, the drug treats anemia by promoting intestinal iron absorption.

[0094] In some embodiments, the drug is used to treat anemia by producing hepcidin.

[0095] Fourthly, this application also provides a health product for invigorating qi and nourishing blood, the ingredients of which include the complex and excipients provided in the second aspect.

[0096] In some embodiments, the excipients comprise at least one of natural, semi-synthetic, and synthetic excipients, such as solvents, propellants, solubilizers, suspending agents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, flow aids, pressure aids, flavoring agents, preservatives, suspending agents, coating agents, and fragrances.

[0097] In some embodiments, the health product may be in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes, or aerosols.

[0098] In some embodiments, the dosage form of the health product includes oral dosage, injection, patch, liniment, spray, inhaler, drops, lotion, or suppository.

[0099] In some embodiments, the health product can promote intestinal iron absorption and inhibit hepcidin, thereby improving iron-deficiency anemia and replenishing blood and qi.

[0100] Fifthly, this application also provides a multifunctional food for improving sub-health conditions, the ingredients of which include the complex and food additives provided in the second aspect.

[0101] The food additives include, but are not limited to, acidity regulators, anti-caking agents, defoamers, antioxidants, bleaching agents, leavening agents, colorants, color protectants, enzyme preparations, flavor enhancers, nutritional fortifiers, preservatives, sweeteners, thickeners, or flavorings.

[0102] For experimental parameters not specified in the following specific embodiments, please refer to the guidelines given in this application document first, or refer to experimental manuals or other experimental methods known in the art, or refer to the experimental conditions recommended by the manufacturer.

[0103] The raw materials and reagents used in the following specific embodiments can be obtained commercially or prepared by those skilled in the art using known methods. Furthermore, the concentrations of each component in the composition refer to the concentration of the corresponding extract component as a whole in the composition or complex. For example, the composition includes 10–20 μg / mL of Cornus officinalis extract, indicating that the amount of Cornus officinalis extract added as a whole in the complex is 10–20 μg of Cornus officinalis extract per 1 mL of the complex. Similarly, the complex includes 10–20 μg / mL of Cornus officinalis extract, indicating that the amount of Cornus officinalis extract added as a whole in the complex is 10–20 μg per 1 mL of the complex.

[0104] Among them: human colon adenocarcinoma Caco-2 cells were purchased from Xiamen Yimo Biotechnology Co., Ltd., China; human liver cancer HepG2 cells were purchased from Cyagen Biosciences Co., Ltd., China; DMEM culture medium and FBS fetal bovine serum were purchased from Gibco, USA; Cell Counting Kit (CCK-8) cell proliferation-toxicity assay kit was purchased from Tongren Chemical Research Institute; Taq Pro Universal SYBR qPCR Master Mix was purchased from Nanjing Novizan Biotechnology Co., Ltd., China; RNA-easy Isolation... Reagent was purchased from Nanjing Novizan Biotechnology Co., Ltd., China; FastKing one-step genomic cDNA first-strand synthesis premix reagent was purchased from Tiangen Biotech (Beijing) Co., Ltd., China; FerroOrange ferrous ion detection probe kit was purchased from Dojin Chemical Research Institute, Japan; 2'-methoxymatrine (CAS: 270249-38-2) and artificially synthesized Panlong phenol-ginseng A (CAS: 126192-35-6) were purchased from WuXi AppTec, Tianjin; Cornus officinalis extract (water-soluble extract of Cornus officinalis pulp, 10:1) and mangosteen extract (water-soluble extract of mangosteen pericarp, 10:1) were both purchased from Fufeng Sinote Biotechnology Co., Ltd.

[0105] Example 1: Application of the composition in the preparation of iron supplements

[0106] The composition formulation provided in this embodiment is as follows:

[0107] Formula 1: 18.2 μg / mL Cornus officinalis extract and 1 μg / mL Mangosteen extract.

[0108] Formula 2: 16 μg / mL Cornus officinalis extract and 3.2 μg / mL Mangosteen extract.

[0109] Formula 3: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract and 0.3 μM 2'-methoxymatrine.

[0110] Formula 4: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract and 0.1 μM Panlongphenol A.

[0111] Formula 5: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine and 0.1 μM Panlongphenol A.

[0112] Formula 6: 14.2 μg / mL Cornus officinalis extract, 5 μg / mL Mangosteen extract, 0.1 μM 2'-methoxymatrine and 0.3 μM Panlongphenol A.

[0113] The concentration data of each component in the above six formulations are the amount added to the composition as a whole for the corresponding extract component.

[0114] The solvent used in the above six formulations is water, which is well known in the art, and the solvent has no effect on the efficacy of the formulation.

[0115] All six of the above formulations have iron-supplementing effects.

[0116] Test Example 1: Cytotoxicity Test of the Composition

[0117] In this experiment, the CCK-8 assay was used to screen for the concentration of the composition that did not significantly inhibit the viability of Caco-2 and HepG2 cells. The specific method is as follows:

[0118] Caco-2 and HepG2 cells in logarithmic growth phase were harvested, digested, and resuspended in complete culture medium. The resulting cell suspension was seeded into 96-well plates at a density of 100 μL per well (1.2 × 10⁶ cells / well). 4 / well, cultured overnight in a 37°C, 5% CO2 incubator. When the cell confluence was about 80%, the following test substances were added: the six formulations 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 Panlongfen A to treat the two cell types.

[0119] The following groups were set up: a blank group containing culture medium and cells without the test substance; a control group containing culture medium and cells without the test substance; and an experimental group containing culture medium and cells containing the test substance. Each group was then incubated at 37°C with 5% CO2 for 24 hours. The original culture medium was then replaced with medium containing 10% CCK-8, and the 96-well plates were placed in an incubator for 4 hours. The OD value of each well was measured at 450 nm. The cell proliferation and survival rate of different test substances was calculated according to formula (I), with three replicates for each concentration.

[0120] The effects of each group on the survival rate of Caco-2 and HepG2 cells obtained according to the above experimental methods are shown in Table 1, Figure 1, and Figure 2:

[0121] Table 1: Effects of the analytes on the survival rates of Caco-2 and HepG2 cells ( n=3)

[0122] The results in Table 1 and Figures 1 and 2 show that the extracts of Cornus officinalis, Mangosteen extract, 2'-methoxymatrine, Panlong phenol ginseng A, and the six combined formulations did not significantly inhibit the survival rate of Caco-2 and HepG2 cells.

[0123] Experimental Example 2: Fe uptake in Caco-2 cells using the composition 2+ Impact

[0124] This experimental method uses a FerroOrange fluorescent probe to detect the effect of the composition on promoting the uptake of iron ions (Fe) by Caco-2 cells. 2+ The specific experimental methods for assessing the impact are as follows:

[0125] Caco-2 cells in the logarithmic growth phase were harvested, digested, and resuspended in complete culture medium. The resulting cell suspension was then seeded into 96-well plates at a density of 100 μL per well (1.2 × 10⁶ cells). 4 / well, cultured in a 37℃, 5% CO2 incubator. When cell confluence was approximately 80%, Caco-2 cells were treated with 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine, 0.1 μM Panlong phenol-ginseng A, and the six formulations provided in Example 1. After 12 h of treatment, the culture medium was discarded, and 100 μL of complete culture medium containing 4 μg / mL ferric citrate and 10 μM ferrous sulfate was added to each well. After overnight culture, the culture medium was discarded, and the cells were washed three times with Hank's salt solution. Then, 1 μM FerroOrange working solution was added, and the cells were incubated in a 7℃, 5% CO2 incubator for 30 min. After incubation, Hoechst 33342 dye was added and incubated for another 5 min. Fluorescence values ​​were detected by instrumentation, and the results are shown in Table 2 and Figure 3.

[0126] Table 2: Effects of analytes on Fe uptake in Caco-2 cells 2+ Impact results Note: Compared to a single substance, * represents p < 0.05, and ** represents p < 0.01;

[0127] The results in Table 2 and Figure 3 show that, compared with the single substance, the six-combination group enhanced the ability of Caco-2 cells to take up iron ions, which is statistically significant.

[0128] Experimental Example 3: Effect of the Composition on Hepcidin mRNA Expression in HepG2 Cells

[0129] This experiment treated HepG2 cells with six different formulations: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine, 0.1 μM Panlongfenshen A, and the formulation provided in Example 1. RNA was extracted, reverse transcribed, and then qPCR was performed to evaluate the mRNA expression level of the Hepcidin gene in HepG2 cells. The specific methods are as follows:

[0130] HepG2 cells in logarithmic growth phase were digested and then suspended in DMEM complete medium containing 15% fetal bovine serum and 1% penicillin-dextrose antibodies. Cells were then seeded at a concentration of 1 × 10⁶ cells / year. 6 Cells were cultured in 6cm cell culture dishes, 5mL per well, in a 37℃, 5% CO2 incubator. After 72h of adaptation culture, the control group was replaced with fresh complete medium, and the intervention group was replaced with complete medium containing the corresponding monomer and composition. After 12h of culture in a 37℃, 5% CO2 incubator, RNA was extracted from each group of cells according to the RNA-easy Isolation Reagent instructions, and the RNA concentration was measured. Following the FastKing one-step genomic cDNA first-strand synthesis premix reagent instructions, the extracted RNA was reverse transcribed into cDNA at a concentration of 100ng / μL. Finally, the experimental system was prepared according to the Taq Pro Universal SYBR qPCR Master Mix instructions, and the mRNA expression level of the Hepcidin gene was detected. The results are shown in Table 3 and Figure 4.

[0131] Table 3: Effects of the analytes on the mRNA expression of the Hepcidin gene Note: Compared to a single substance, * indicates p < 0.05;

[0132] The results in Table 3 and Figure 4 show that, compared with the single substance, the six-component combination significantly reduced the mRNA expression of Hepcidin, which was statistically significant.

[0133] Experimental Example 4: Effect of the Composition on Hepcidin Protein Expression in HepG2 Cells

[0134] This experiment treated HepG2 cells with six different formulations: 16 μg / mL Cornus officinalis extract, 3.2 μg / mL Mangosteen extract, 0.3 μM 2'-methoxymatrine, 0.1 μM Panlongfenshen A, and the formulation provided in Example 1. After protein extraction, Western blot experiments were performed to evaluate the expression level of Hepcidin protein in HepG2 cells. The specific methods are as follows:

[0135] HepG2 cells in logarithmic growth phase were digested and then suspended in DMEM complete medium containing 10% fetal bovine serum and 1% penicillin-dextrose antibodies. Cells were then seeded at a concentration of 1 × 10⁶ cells / year. 6Cells were cultured in 6cm cell culture dishes, 5mL per well, at 37℃ in a 5% CO2 incubator. After 72h of adaptation culture, the control group was replaced with fresh complete medium, while the intervention group was replaced with complete medium containing the aforementioned monomers and mixtures. After 12h of culture at 37℃ in a 5% CO2 incubator, cells were scraped off with a cell scraper, centrifuged at 1000rpm for 5min, lysed on ice, and total protein was extracted. Protein concentration was determined using a BCA kit. 30μg of protein from each group was subjected to SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% BSA, and incubated overnight at 4℃ with Hepcidin and GAPDH primary antibodies. The membranes were washed with TBS-T washing buffer, incubated at room temperature for 1h with HRP-labeled secondary antibody, washed with TBS-T washing buffer, and photographed using a chemiluminescence imaging system. The data are shown in Table 4 and Figure 5.

[0136] Table 4: Effects of the analytes on Hepcidin protein expression Note: Compared to a single substance, * indicates p < 0.05;

[0137] The results in Table 4 and Figure 5 show that, compared with the single substance, the six combinations can significantly reduce the protein expression of Hepcidin, which is statistically significant.

[0138] Data processing was performed using GraphPadPrism 9.3.0 statistical software. Some experimental results are expressed as Mean ± SD.

[0139] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above 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 embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. The use of the composition in the preparation of iron supplements, wherein the composition comprises Cornus officinalis and mangosteen, wherein the Cornus officinalis comprises at least one of Cornus officinalis medicinal material and Cornus officinalis extract, and the mangosteen comprises at least one of mangosteen medicinal material and mangosteen extract.

2. The application according to claim 1, wherein, The composition satisfies at least one of the following (1) to (3): (1) The mass ratio of Cornus officinalis and Mangosteen in the composition is (10-20):(1-5); (2) The Cornus officinalis extract is selected from at least one of water extract and alcohol extract; (3) The mangosteen extract is selected from at least one of water extract and alcohol extract.

3. The application according to claim 1 or 2, wherein, The composition also includes at least one of 2'-methoxymatrine and Panlongphenol A.

4. The application according to claim 3, wherein, The composition satisfies at least one of the following (1) and (2): (1) The amount of 2'-methoxymatrine added to the composition is 0.1-1 μM; (2) The amount of Panlong phenol A added in the composition is 0.05-0.5 μM.

5. The composition according to claim 4, wherein, 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 Panlongphenol A.

6. The application according to claim 1 or 2, wherein, The iron supplement works by promoting intestinal iron absorption.

7. The application according to claim 1 or 2, wherein, The iron supplement works by inhibiting hepcidin.

8. A complex comprising Cornus officinalis and mangosteen, wherein the Cornus officinalis comprises at least one of Cornus officinalis medicinal material and Cornus officinalis extract, and the mangosteen comprises at least one of mangosteen medicinal material and mangosteen extract, and further comprises at least one of 2'-methoxymatrine and Panlongphenol A.

9. The complex according to claim 8, wherein, The complex satisfies at least one of the following (1) to (3): (1) The content ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the complex is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM; (2) The mass ratio of Cornus officinalis, Mangosteen and Panlong phenolic A in the complex 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 Panlong phenol A in the complex is (10-20) μg / mL: (1-5) μg / mL: (0.1-1) μM: (0.05-0.5) μM.

10. The complex according to claim 8 or 9, wherein, The complex satisfies at least one of the following (1) to (5): (1) The mass ratio of Cornus officinalis, Mangosteen and 2'-methoxymatrine in the complex is (15-18) μg / mL: (3-4) μg / mL: (0.1-0.5) μM; (2) The mass ratio of Cornus officinalis, Mangosteen and Panlong phenolic A in the complex 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 Panlong phenolic A in the complex 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 at least one of water extract and alcohol extract; (5) The mangosteen extract is selected from at least one of water extract and alcohol extract.

11. The complex according to claim 10, wherein, The complex 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 Panlongphenol A.

12. A medicament for treating anemia, the medicament comprising the complex according to any one of claims 8 to 11, and pharmaceutically acceptable excipients.

13. The drug for treating anemia according to claim 12, wherein, The drug satisfies at least one of the following (1) to (5): (1) The form of the drug includes tablets, granules, capsules, solutions, powders, suspensions, emulsions, ointments or aerosols; (2) The dosage forms of the drug include oral preparations, injections, patches, liniments, sprays, inhalers, drops, lotions, or suppositories; (3) The anemia mentioned is iron deficiency anemia, autoimmune hemolytic anemia, or nausea anemia; (4) The drug treats anemia by promoting intestinal iron absorption; (5) The drug treats anemia by inhibiting hepcidin.

14. A health product for invigorating qi and nourishing blood, wherein the ingredients of the health product include the complex and excipients described in any one of claims 8 to 11.

15. The health product for invigorating qi and nourishing blood according to claim 14, wherein, The health products can be in the form of tablets, granules, capsules, solutions, powders, suspensions, emulsions, pastes, or aerosols.

16. A multifunctional food for improving sub-health conditions, the food comprising the complex according to any one of claims 8 to 11 and food additives.

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