Use of 18β-glycyrrhetinic acid for improving hematopoietic ability of renal anemia

Abstract

The present invention provides the use of 18β-glycyrrhetinic acid for improving the hematopoietic capacity of renal anemia. The present invention discovers and verifies for the first time that 18β-glycyrrhetinic acid stimulates the production of endogenous EPO, thereby improving the body's hematopoietic capacity. Short-term supplementation of 18β-glycyrrhetinic acid can significantly increase the level of erythropoietin, thereby promoting the production of reticulocytes. Long-term supplementation of 18β-glycyrrhetinic acid can improve indicators such as red blood cell count and alleviate the symptoms of renal anemia. The present invention provides a natural, safe, and highly effective active ingredient that improves the body's hematopoietic capacity, provides a scientific basis for basic research on drugs for the clinical treatment and prevention of renal anemia, and has significant application value.

Description

Technical Field

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

[0002] Renal anemia is a type of hypoproliferative anemia caused by impaired renal function, typically characterized by decreased red blood cell (RBC) count, hematocrit (HCT), or hemoglobin (HGB) content in the peripheral blood. The incidence of anemia increases with declining renal function. Insufficient endogenous erythropoietin (EPO) secretion, resulting from renal tissue lesions, is a major cause of renal anemia. EPO is the primary growth factor regulating erythropoiesis. During embryonic development, EPO is primarily synthesized in the liver and, after birth, is produced in the renal cortex interstitial cells. Currently, recombinant human EPO (rhEPO) is the most commonly used erythropoiesis-stimulating agent in clinical practice. However, its injection may cause off-target effects, increasing the risk of cardiovascular disease, hypertension, and stroke. 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), an oleanane-type pentacyclic triterpenoid compound and the main active ingredient in licorice root, is widely used in the modern food industry as a sweetener and flavoring. Numerous studies have demonstrated that 18β-GA possesses antioxidant, anti-inflammatory, anti-cancer, and hepatoprotective properties. Patent publication number CN111758963A discloses that 18β-GA can rescue myeloid differentiation in bone marrow hematopoiesis and alleviate aging of bone marrow hematopoietic stem cells in an aging model, with the mechanism of action possibly related to its anti-inflammatory properties. However, there are currently no reports on the therapeutic effects of 18β-GA 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:

[0006] In a first aspect, the present invention provides use of 18β-glycyrrhetinic acid in preparing a product for treating renal anemia.

[0007] Furthermore, the structural formula of the 18β-glycyrrhetinic acid (CAS: 471-53-4) is as follows:

[0008] .

[0009] Furthermore, the product is a medicine.

[0010] Furthermore, the application includes one of the following:

[0011] (1) Application in the preparation of products for increasing endogenous erythropoietin levels;

[0012] (2) Application in the preparation of products for promoting reticulocyte production;

[0013] (3) Application in the preparation of products for increasing the number of red blood cells;

[0014] (4) Application in the preparation of products for increasing hemoglobin content;

[0015] (5) Application in the preparation of products for increasing hematocrit;

[0016] (6) Application in the preparation of products for improving hematopoietic capacity.

[0017] Furthermore, the effect of 18β-glycyrrhetinic acid in improving the hematopoietic capacity of renal anemia is manifested as follows:

[0018] (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;

[0019] (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;

[0020] (3) Experiments have shown that taking 18β-glycyrrhetinic acid for 3 weeks can effectively increase the number of red blood cells, hemoglobin content and hematocrit of anemic mice.

[0021] Furthermore, after short-term use (1 week), it has the effect of promoting RET production, and after 3 weeks of use, it can increase the red blood cell and hemoglobin levels of anemic mice and alleviate anemia symptoms.

[0022] Furthermore, the drug is prepared using 18β-glycyrrhetinic acid as the active ingredient, pharmaceutically or physiologically acceptable excipients, and conventional pharmaceutical preparation technology.

[0023] Furthermore, the medicine is an oral preparation or an injection.

[0024] Furthermore, the oral preparation is a powder, granule, capsule, powder, pill, tablet or oral liquid.

[0025] Furthermore, the renal anemia is anemia caused by reduced erythropoietin production when kidney disease leads to decreased renal function.

[0026] 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.

[0027] 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.

[0028] Furthermore, the 18β-glycyrrhetinic acid is the only active ingredient.

[0029] Furthermore, the daily dosage of 18β-glycyrrhetinic acid does not exceed 10 mg / kg; preferably 2-8 mg / kg; further 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 dosages can vary according to the needs of the patient, the severity of the condition being treated, and the compound used. Generally, 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 divided into divided doses within a day if necessary.

[0030] 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.

[0031] 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.

[0032] For the preparation of medicaments suitable for the present invention, pharmaceutically acceptable carriers can be either solid or liquid.

[0033] Solid form medicines include powders, granules, capsules, powders, pills, and tablets. Solid carriers can be one or more substances that also act as diluents, flavorings, solubilizers, lubricants, suspending agents, adhesives, preservatives, tablet disintegrating agents, or encapsulating materials. In powders, the carrier is a finely divided solid that is mixed with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary adhesive properties in an appropriate ratio and compressed into the 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.

[0034] 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.

[0035] Therefore, the medicaments 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 presented in the form of a unit dose in ampoules, prefilled syringes, small-volume infusion bags, or multi-dose containers together with added preservatives. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle 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 isolation of a sterilized solid or lyophilization from a solution, for reconstitution with a suitable carrier such as sterile, pyrogen-free water before use.

[0036] Aqueous solutions suitable for oral administration can be prepared by dissolving the active ingredient in water and adding colorants, flavorings, stabilizers, and thickeners as desired. 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 well-known suspending agents.

[0037] Also included are solid dosage forms 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, these drugs may contain colorants, flavorings, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, etc.

[0038] Compared with the prior art, the novel use of 18β-glycyrrhetinic acid for improving the hematopoietic capacity of renal anemia according to the present invention has the following advantages:

[0039] The present invention discovered and verified for the first time that 18β-glycyrrhetinic acid stimulates the production of endogenous EPO, thereby improving the body's hematopoietic ability and improving the symptoms of renal anemia. It provides a natural, safe, and highly effective active ingredient that can improve the body's hematopoietic ability, provides a scientific basis for basic research on drugs for the clinical treatment and prevention of renal anemia, and has significant application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The accompanying drawings, which constitute part of the present invention, are provided to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are provided to explain the present invention and do not constitute an undue limitation of the present invention. In the accompanying drawings:

[0041] 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;

[0042] 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;

[0043] 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;

[0044] 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;

[0045] 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

[0046] 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.

[0047] The present invention will be described in detail below with reference to the accompanying drawings and in conjunction with embodiments.

[0048] Example 1 Effect of short-term supplementation of 18β-glycyrrhetinic acid on the hematopoietic ability of renal anemia model mice

[0049] Experimental animal groups: 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).

[0050] Renal anemia model: A renal anemia model was induced in mice using an adenine-rich diet. The model group and the model treatment group were fed a standard rodent maintenance diet containing 0.2% adenine, while the control group and the treatment group were fed the corresponding normal diet for 4 weeks.

[0051] Experimental method: The gavage method was used. Starting from the third week of modeling, the control group and the model group were 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 of 18β-glycyrrhetinic acid, with the solvent being 0.5% sodium carboxymethyl cellulose, every day for 1 week.

[0052] After the experiment, whole blood was collected from the mice using eye sampling and aliquoted into 1.5 ml centrifuge tubes and anticoagulant tubes (coated with EDTA K2). Approximately 200 μL of the mouse's whole blood was dripped into the anticoagulant tube and immediately analyzed using a fully automated animal hematology analyzer. The remaining whole blood was dripped into a 1.5 ml centrifuge tube and allowed to stand at room temperature for 2 hours before centrifugation to collect serum for testing.

[0053] Serum EPO content determination: Serum EPO levels were determined using a mouse EPO enzyme-linked immunosorbent assay (ELISA) kit.

[0054] like Figure 1 As shown in the results, the proportion and absolute value of reticulocytes in the Ad group were significantly lower than those in the Con group (P<0.0001), while the proportion and absolute value of reticulocytes in the Ad+GA group increased by approximately 67% and 83%, respectively, compared with the Ad group (P<0.0001), indicating that 18β-glycyrrhetinic acid has the potential to improve the hematopoietic ability of mice with renal anemia model.

[0055] like Figure 2 As shown in the data, the serum EPO level in the C+GA group increased by 67% compared with the Con group (P<0.05). In addition, the serum EPO level in the Ad+GA group further increased significantly compared with the Ad group (P<0.01), and the serum EPO level in the Ad group also increased significantly compared with the Con group (P<0.001). This indicates that after the hematopoietic function of the Ad group mice was damaged, the level of EPO secretion in their kidneys showed a compensatory increase.

[0056] 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.

[0057] Example 2 Evaluation of the effect of long-term supplementation of 18β-glycyrrhetinic acid on renal anemia model mice

[0058] Experimental animal groups: 48 male C57BL / 6J mice, 6-8 weeks old, were randomly divided into 6 groups, with 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).

[0059] Renal Anemia Model: A renal anemia model was established in mice using an adenine-rich diet. The model group, model / drug group 1, and model / drug group 2 were fed a standard rodent maintenance diet containing 0.2% adenine for 4 weeks, followed by a switch to a normal maintenance diet without adenine for 3 weeks. The control group and control / drug groups 1 and 2 were fed the corresponding normal diet for 7 weeks.

[0060] Experimental Methods: Starting from the fourth week of modeling, the control and model groups were gavaged daily with 0.5% sodium carboxymethylcellulose for three weeks. Control / drug group 1 and model / drug group 1 were gavaged daily with 25 mg / kg of 18β-glycyrrhetinic acid in 0.5% sodium carboxymethylcellulose for three weeks. Control / drug group 2 and model / drug group 2 were gavaged daily with 50 mg / kg of 18β-glycyrrhetinic acid for three weeks. At the end of the experiment, whole blood was collected by eye sampling and aliquoted into 1.5 ml centrifuge tubes and anticoagulant tubes (coated with EDTA K2). Approximately 200 μL of whole blood was dripped into the anticoagulant tube and immediately analyzed using an automated animal hematology analyzer. The remaining whole blood was dripped into a 1.5 ml centrifuge tube, allowed to stand at room temperature for 2 hours, and then centrifuged to obtain serum for analysis.

[0061] Serum EPO content determination: Serum EPO levels were determined using a mouse EPO enzyme-linked immunosorbent assay (ELISA) kit.

[0062] 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). However, 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.

[0063] like Figure 4As shown, the proportion and absolute value of reticulocytes in the Ad group increased by approximately 34% (P < 0.01) and 23% (P < 0.05), respectively, compared with the Con group. This suggests that the 3-week maintenance diet period after model establishment resulted in a compensatory increase in RET levels, possibly due to the enhanced stimulation of EPO on erythropoiesis. Supplementation with 18β-glycyrrhetinic acid restored RET levels to almost normal levels.

[0064] like Figure 5 As shown in the results, long-term supplementation of 18β-glycyrrhetinic acid under normal conditions 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 the potential to treat renal anemia.

[0065] The above results show that in the renal anemia mouse model, long-term supplementation of 18β-glycyrrhetinic acid can improve indicators such as red blood cell count, alleviate the symptoms of renal anemia in mice, and play a role in treating renal anemia.

[0066] The embodiments described above are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work shall fall within the scope of protection of the present invention.

Claims

1. Use of 18β-glycyrrhetinic acid in the preparation of a medicament for treating renal anemia, wherein the renal anemia is anemia caused by reduced erythropoietin production when renal disease leads to decreased renal function, wherein the 18β-glycyrrhetinic acid is the sole active ingredient of the medicament, and the renal anemia is renal anemia due to diabetic kidney disease, renal anemia complicated by chronic kidney disease, or renal anemia complicated by cancer.

2. The use according to claim 1, characterized in that The 18β-glycyrrhetinic acid increases endogenous erythropoietin levels.

3. The use according to claim 1 or 2, characterized in that: The drug is prepared using 18β-glycyrrhetinic acid as an active ingredient, pharmaceutically or physiologically acceptable excipients, and a conventional drug preparation process.

4. The use according to claim 3, characterized in that: The medicine is an oral preparation or an injection.

5. The use according to claim 4, characterized in that: The oral preparation is a powder, granule, capsule, powder, pill, tablet or oral liquid.

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