New application of 18 beta-glycyrrhetinic acid in improving hematopoietic capacity of renal anemia

Abstract

The invention provides a new application of 18beta-glycyrrhetinic acid in improving hematopoietic capacity of renal anemia, and finds and verifies that the hematopoietic capacity of an organism is improved by stimulating generation of endogenous EPO (erythropoietin) through 18beta-glycyrrhetinic acid for the first time, and the level of erythropoietin can be remarkably increased by supplementing 18beta-glycyrrhetinic acid for a short time; by supplementing the 18 beta-glycyrrhetinic acid for a long time, indexes such as the number of red blood cells can be improved, and the symptom of renal anemia can be relieved. The invention provides an active component which is natural, safe and efficient and can improve the hematopoietic capacity of the organism, provides a scientific basis for the fundamental research of medicines for clinically treating and preventing renal anemia and the development of health-care foods, and has important application value.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine, and in particular relates to a new use of 18β-glycyrrhetinic acid for improving the hematopoietic ability of renal anemia. Background Art

[0002] Renal anemia is a type of hypoproliferative anemia caused by impaired renal function, that is, anemia caused by decreased red blood cell production. Renal anemia is one of the common complications of patients with chronic kidney disease. It is usually manifested as a decrease in red blood cell count (RBC), hematocrit (HCT) or hemoglobin (HGB) content in peripheral blood. As renal function declines, the incidence of anemia gradually increases. Insufficient secretion of endogenous erythropoietin (EPO) caused by renal tissue lesions is one of the main causes of renal anemia. EPO is the main growth factor that regulates the body's red blood cell production. EPO is mainly synthesized in the liver during the embryonic period and produced by the interstitial cells of the renal cortex layer of the kidney after birth. At present, recombinant human EPO (rhEPO) is the most commonly used erythropoiesis stimulator in clinical practice, but its injection may cause off-target effects and increase the risk of cardiovascular disease, hypertension, and stroke in patients. Therefore, it is of great significance to find and explore an active substance that can promote the production of endogenous EPO and thus improve the hematopoietic ability of the kidney.

[0003] 18β-Glycyrrhetinic acid (GA) is an oleanane-type pentacyclic triterpenoid compound and the main active ingredient of licorice root. It is widely used as a food sweetener and flavoring agent in the modern food industry. A large number of studies have shown that 18β-glycyrrhetinic acid has biological functions such as antioxidant, anti-inflammatory, anti-cancer, and liver protection. The patent with publication number CN111758963A discloses that 18β-glycyrrhetinic acid can rescue the myeloid differentiation tendency of bone marrow hematopoiesis in an aging model and alleviate the aging of bone marrow hematopoietic stem cells. Its mechanism of action may be related to its anti-inflammatory effect. At present, there is no report on the therapeutic effect of 18β-glycyrrhetinic acid on renal anemia. Summary of the invention

[0004] In view of this, the present invention aims to propose a new use of 18β-glycyrrhetinic acid for improving the hematopoietic ability of renal anemia. Experiments have found that 18β-glycyrrhetinic acid can improve the body's hematopoietic ability by inducing the production of endogenous EPO, thereby improving renal anemia.

[0005] To achieve the above object, the technical solution of the present invention is achieved as follows: In a first aspect, the present invention provides use of 18β-glycyrrhetinic acid in preparing a product for treating renal anemia.

[0006] Furthermore, the structural formula of the 18β-glycyrrhetinic acid (CAS: 471-53-4) is as follows: .

[0007] Furthermore, the product is one of a medicine, a health product, and a functional food.

[0008] Furthermore, the application includes one of the following: (1) Application in the preparation of products for increasing endogenous erythropoietin levels; (2) Application in the preparation of products for promoting reticulocyte production; (3) Application in the preparation of products for increasing the number of red blood cells; (4) Application in the preparation of products for increasing hemoglobin content; (5) Application in the preparation of products for increasing hematocrit; (6) Application in the preparation of products for improving hematopoietic capacity.

[0009] Furthermore, the effect of 18β-glycyrrhetinic acid in improving the hematopoietic capacity of renal anemia is as follows: (1) The results of various experiments on the intervention of 18β-glycyrrhetinic acid in renal anemia model mice showed that 18β-glycyrrhetinic acid can effectively increase the serum EPO level of anemic mice and has the potential to alleviate anemia symptoms; (2) By analyzing the changes in the level of reticulocytes (RET), a marker of bone marrow hematopoietic function, it was found that 18β-glycyrrhetinic acid can significantly promote the production of RET and restore it to normal levels; (3) Experiments have shown that taking 18β-glycyrrhizic acid for 3 weeks can effectively increase the number of red blood cells, hemoglobin content and hematocrit of anemic mice.

[0010] Furthermore, after short-term use (1 week), it has the effect of promoting RET production, and taking it for 3 weeks can increase the red blood cell and hemoglobin levels of anemic mice and alleviate anemia symptoms.

[0011] Furthermore, the drug is prepared with 18β-glycyrrhetinic acid as the active ingredient, pharmaceutically or physiologically acceptable excipients and conventional drug preparation processes.

[0012] Furthermore, the medicine is an oral preparation or an injection.

[0013] Furthermore, the oral preparation is a powder, granule, capsule, powder, pill, tablet or oral liquid.

[0014] Furthermore, the renal anemia is anemia caused by reduced erythropoietin production when kidney disease leads to decreased renal function.

[0015] Furthermore, the renal anemia is renal anemia caused by diabetic kidney disease, renal anemia complicated by chronic kidney disease, or renal anemia complicated by cancer.

[0016] In a second aspect, the present invention provides a drug for treating renal anemia, wherein the active ingredient of the drug includes 18β-glycyrrhetinic acid.

[0017] Furthermore, the 18β-glycyrrhetinic acid is the only active ingredient.

[0018] Further, the daily dosage of 18β-glycyrrhetinic acid does not exceed 10 mg / kg; preferably 2-8 mg / kg; more preferably 3-5 mg / kg, for example, 3 mg / kg, 3.2 mg / kg, 3.4 mg / kg, 3.5 mg / kg, 3.6 mg / kg, 3.8 mg / kg, 4 mg / kg, 4.2 mg / kg, 4.4 mg / kg, 4.5 mg / kg, 4.6 mg / kg, 4.8 mg / kg, 5 mg / kg. These doses can vary according to the needs of the patient, the severity of the disease being treated, and the compound used. Generally speaking, treatment is started with a smaller dose that is less than the optimal dose of the compound, and thereafter, the dose is increased in small amounts to achieve the best effect. For convenience, the total daily dose can be further subdivided into divided doses within a day if necessary.

[0019] Furthermore, the drug also includes pharmaceutically or physiologically acceptable excipients, which include one or more of diluents, adhesives, wetting agents, lubricants, disintegrants, solvents, emulsifiers, solubilizers, preservatives, pH regulators, osmotic pressure regulators, surfactants, coating materials, antioxidants or buffers.

[0020] Furthermore, the administration route of the drug includes at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, and sublingual administration.

[0021] For the preparation of suitable medicaments of the present invention, pharmaceutically acceptable carriers may be solid or liquid.

[0022] Solid form medicines include powders, granules, capsules, powders, pills, tablets. Solid carriers can be one or more substances that also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, adhesives, preservatives, tablet disintegrating agents, or encapsulating materials. In powders, the carrier is a subdivided solid that is mixed with the subdivided active ingredient. In tablets, the active ingredient is mixed with a carrier with the necessary adhesive properties in an appropriate ratio and compressed into a desired shape and size. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, etc.

[0023] Oral liquids include solutions, suspensions and emulsions, for example, aqueous solutions or water-propylene glycol solutions. For example, parenteral injection liquid preparations can be formulated as water-polyethylene glycol solutions.

[0024] Therefore, the medicament used in the present invention can be formulated into a preparation for parenteral administration (e.g., injection, such as bolus injection or continuous infusion), and can be present in the form of a unit dose in an ampoule, a prefilled syringe, a small volume infusion bag or a multi-dose container together with an added preservative. The composition can take the form of a suspension, solution or emulsion of an oily or aqueous carrier, and can contain formulation ingredients such as suspending agents, stabilizers and / or dispersants. In addition, the active ingredient can be in the form of a powder, which can be obtained by aseptic separation of a sterilized solid or freeze-drying from a solution, for reconstitution with a suitable carrier such as sterile, pyrogen-free water before use.

[0025] Aqueous solutions suitable for oral administration can be prepared by dissolving the active ingredient in water and adding desired colorants, flavoring agents, stabilizers and thickeners. Aqueous suspensions suitable for oral administration can be prepared by dispersing the finely divided active ingredient in water containing a viscous material, such as a natural or synthetic gum, resin, methylcellulose, sodium carboxymethylcellulose, or other known suspending agents.

[0026] Also included are solid preparations designed to be converted into liquid preparations for oral administration shortly before use. Such liquid preparations include solutions, suspensions and emulsions. In addition to the active ingredient, such drugs may contain colorants, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, etc.

[0027] Compared with the prior art, the new use of 18β-glycyrrhetinic acid for improving the hematopoietic capacity of renal anemia according to the present invention has the following advantages: The present invention discovers and verifies for the first time that 18β-glycyrrhetinic acid stimulates the production of endogenous EPO and thus improves the body's hematopoietic ability and improves the symptoms of renal anemia, thereby providing a natural, safe, and highly effective active ingredient that can improve the body's hematopoietic ability, providing a scientific basis for basic research on clinical drugs for the treatment and prevention of renal anemia and the development of health foods, and having great application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the accompanying drawings: Figure 1 This is a schematic diagram of the effect of short-term supplementation of 18β-glycyrrhetinic acid on mouse reticulocytes (RET) as described in Example 1 of the present invention; Figure 2 This is a schematic diagram of the effect of short-term supplementation of 18β-glycyrrhetinic acid on erythropoietin (EPO) in mice as described in Example 1 of the present invention; Figure 3 This is a schematic diagram of the changes in anemia indicators such as red blood cell count (RBC), hemoglobin (HGB) and hematocrit (HCT) in mice after long-term supplementation with 18β-glycyrrhetinic acid as described in Example 2 of the present invention; Figure 4 This is a schematic diagram of the effect of long-term supplementation of 18β-glycyrrhetinic acid on mouse reticulocytes (RET) as described in Example 2 of the present invention; Figure 5 This is a schematic diagram of the effect of long-term supplementation of 18β-glycyrrhetinic acid on erythropoietin (EPO) in mice as described in Example 2 of the present invention. DETAILED DESCRIPTION

[0029] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0030] The present invention will be described in detail below with reference to the accompanying drawings and in conjunction with embodiments.

[0031] Example 1 Effect of short-term supplementation of 18β-glycyrrhetinic acid on the hematopoietic ability of renal anemia model mice Experimental animal grouping: 32 male C57BL / 6J mice, 6-8 weeks old, were randomly divided into 4 groups, 8 mice in each group, namely control group (Con), drug administration group (C+GA), model group (Ad) and model drug administration group (Ad+GA).

[0032] Renal anemia model: Adenine diet was used to induce a renal anemia model in mice. The model group and the model treatment group were fed with a standard rodent maintenance diet containing 0.2% adenine, and the control group and the treatment group were fed with the corresponding normal diet for 4 weeks.

[0033] Experimental method: The gavage method was used. From the third week of modeling, the control group and the model group began to be gavaged with 0.5% sodium carboxymethyl cellulose every day for 1 week. The drug-treated group and the model-treated group were gavaged with 100 mg / kg 18β-glycyrrhetinic acid every day, with the solvent being 0.5% sodium carboxymethyl cellulose, for 1 week.

[0034] After the experiment, whole blood was collected by eye sampling and divided into 1.5 ml centrifuge tubes and anticoagulant tubes (coated with EDTA K2). About 200 μL of whole blood was dripped into the anticoagulant tube, and blood routine tests were immediately performed using a fully automatic animal blood analyzer. The remaining whole blood was dripped into a 1.5 ml centrifuge tube, placed at room temperature for 2 hours, and then centrifuged to obtain serum for testing.

[0035] Serum EPO content determination: Serum EPO levels were determined using mouse EPO enzyme-linked immunosorbent assay (ELISA).

[0036] like Figure 1 As shown in the data, the proportion and absolute value of reticulocytes in the Ad group were significantly lower than those in the Con group (P<0.0001), while compared with the Ad group, the proportion and absolute value of reticulocytes in the Ad+GA group increased by approximately 67% and 83%, respectively (P<0.0001), indicating that 18β-glycyrrhizic acid has the potential to improve the hematopoietic ability of mice with renal anemia model.

[0037] like Figure 2 As shown, the serum EPO level in the C+GA group increased by 67% compared with the Con group (P<0.05). In addition, compared with the Ad group, the serum EPO level in the Ad+GA group increased significantly (P<0.01), and the serum EPO level in the Ad group was also significantly increased compared with the Con group (P<0.001). This indicates that after the hematopoietic function of the mice in the Ad group was damaged, the level of EPO secretion in their kidneys showed a compensatory increase.

[0038] In summary, in the renal anemia mouse model, short-term supplementation of 18β-glycyrrhetinic acid can improve the body's hematopoietic capacity by stimulating the production of endogenous EPO.

[0039] Example 2 Evaluation of the effect of long-term supplementation of 18β-glycyrrhetinic acid in the treatment of renal anemia model mice Experimental animal grouping: 48 male C57BL / 6J mice, 6-8 weeks old, were randomly divided into 6 groups, 8 mice in each group, namely control group (Con), control / drug group 1 (C+GA25), control / drug group 2 (C+GA50), model group (Ad), model / drug group 1 (Ad+GA25) and model / drug group 2 (Ad+GA50).

[0040] Renal anemia model: Adenine diet was used to induce renal anemia in mice. The model group, model / drug group 1, and model / drug group 2 were fed with a standard rodent maintenance diet containing 0.2% adenine for 4 weeks, and then switched to a normal maintenance diet without adenine for 3 weeks. The control group, control / drug groups 1 and 2 were fed with the corresponding normal diet for 7 weeks.

[0041] Experimental methods: The control group and the model group were gavaged with 0.5% sodium carboxymethyl cellulose every day from the 4th week of modeling for 3 weeks; the control / drug group 1 and the model / drug group 1 were gavaged with 25 mg / kg of 18β-glycyrrhetinic acid every day, with the solvent being 0.5% sodium carboxymethyl cellulose, for 3 weeks; the corresponding control / drug group 2 and the model / drug group 2 were gavaged with 50 mg / kg of 18β-glycyrrhetinic acid every day for 3 weeks. After the experiment, whole blood was collected by eyeball blood sampling, and the blood of the mice was divided into 1.5 ml centrifuge tubes and anticoagulant tubes (coated with EDTA K2). About 200 μL of whole blood of the mice was dripped into the anticoagulant tube, and blood routine tests were immediately performed using an automatic animal blood analyzer. The remaining whole blood was dripped into a 1.5 ml centrifuge tube, and the serum was centrifuged and collected after being placed at room temperature for 2 hours for testing.

[0042] Serum EPO content determination: Serum EPO levels were determined using mouse EPO enzyme-linked immunosorbent assay (ELISA).

[0043] like Figure 3 As shown, the three key indicators of anemia symptoms, red blood cell count (RBC), hemoglobin (HGB) and hematocrit (HCT), in the Ad group were significantly lower than those in the Con group (P<0.0001), while continuous supplementation of 18β-glycyrrhetinic acid for 3 weeks significantly increased the levels of RBC, HGB and HCT in mice with renal anemia in a dose-dependent manner, indicating that 18β-glycyrrhetinic acid can improve anemia.

[0044] like Figure 4As shown in the figure, the proportion and absolute value of reticulocytes in the Ad group increased by about 34% (P<0.01) and 23% (P<0.05) respectively compared with the Con group, which indicates that the 3-week maintenance diet period after the modeling caused a compensatory increase in RET levels, and its mechanism may be related to the enhanced stimulation of EPO on erythropoiesis. After supplementation with 18β-glycyrrhetinic acid, RET almost returned to normal levels.

[0045] like Figure 5 As shown in the results, under normal conditions, long-term supplementation of 18β-glycyrrhetinic acid can increase serum EPO levels, but the effect is not obvious. In the renal anemia model, serum EPO levels of Ad+GA25 and Ad+GA50 increased by 48% (P<0.05) and 81% (P<0.05) respectively compared with the Ad group, further indicating that 18β-glycyrrhetinic acid has a potential role in treating renal anemia.

[0046] The above results show that in the renal anemia mouse model, long-term supplementation of 18β-glycyrrhetinic acid can improve indicators such as the number of red blood cells, alleviate the symptoms of renal anemia in mice, and play a role in treating renal anemia.

[0047] The embodiments described above are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work shall all fall within the scope of protection of the present invention.

Claims

1. 18 Use of β-glycyrrhetinic acid in the preparation of products for treating renal anemia.

2. The use according to claim 1, characterized in that: The product is one of medicine, health care product and functional food.

3. The use according to claim 1, characterized in that: The application includes at least one of the following: (1) Application in the preparation of products for increasing endogenous erythropoietin levels; (2) Application in the preparation of products for promoting reticulocyte production; (3) Application in the preparation of products for increasing the number of red blood cells; (4) Application in the preparation of products for increasing hemoglobin content; (5) Application in the preparation of products for increasing hematocrit; (6) Application in the preparation of products for improving hematopoietic capacity.

4. The use according to claim 2, characterized in that: The drug is prepared by using 18β-glycyrrhetinic acid as an active component, pharmaceutically or physiologically acceptable auxiliary materials and a conventional drug preparation process.

5. The use according to claim 4, characterized in that: The medicine is an oral preparation or an injection.

6. The use according to claim 5, characterized in that: The oral preparation is a powder, granule, capsule, powder, pill, tablet or oral liquid.

7. The use according to claim 1, characterized in that: The renal anemia is anemia caused by decreased production of erythropoietin when kidney disease leads to decreased renal function.

8. The use according to claim 1, characterized in that: The renal anemia is renal anemia due to diabetic kidney disease, renal anemia complicated by chronic kidney disease, or renal anemia complicated by cancer.

9. A drug for treating renal anemia, characterized in that: The active ingredient of the drug includes 18β-glycyrrhetinic acid.

10. The drug according to claim 9, characterized in that: The drug also includes pharmaceutically or physiologically acceptable excipients, which include one or more of diluents, adhesives, wetting agents, lubricants, disintegrants, solvents, emulsifiers, solubilizers, preservatives, pH regulators, osmotic pressure regulators, surfactants, coating materials, antioxidants or buffers.

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