Application of eplerenone in preparation of medicine for treating type 1 diabetes

Abstract

The invention provides an application of eplerenone in preparation of a medicine for treating type 1 diabetes mellitus. The application finds that eplerenone can be used for effectively treating type 1 diabetes mellitus: reducing the blood glucose level of a type 1 diabetes mellitus model mouse, increasing the serum insulin level and the serum C peptide level, and improving the survival rate of the mouse.

Description

Technical Field

[0001] This application belongs to the field of the treatment of autoimmune diseases, particularly type 1 diabetes. Specifically, this application provides the use of eplerenone in the preparation of a medicament for treating type 1 diabetes. Background Art

[0002] Type 1 diabetes mellitus (T1DM) is an endocrine and metabolic disease caused by the combined action of susceptible genes and environmental factors, resulting in the destruction of pancreatic islet β cells and lifelong dependence on insulin therapy.

[0003] T1DM and type 2 diabetes mellitus (T2DM) are two distinct diseases.

[0004] T1DM is an autoimmune disease that occurs when the body's immune system mistakenly regards insulin-producing β cells as foreign invaders. Risk factors include genetics, age, and environment. It usually has a sudden onset, and many patients develop symptoms within a few weeks. It can occur at any age, most commonly starting at a very young age, but it does not exclude occurrence in adults. Common symptoms include hyperglycemia within a short period, leading to weight loss, fatigue, thirst, and frequent urination. Short-term complications include hypoglycemia and ketoacidosis. In the long term, it may cause damage to blood vessels, affect the heart and kidneys, and lead to premature death.

[0005] T2DM is a metabolic disease in which the pancreas produces sufficient insulin, but the body cannot utilize it. Risk factors include age, exercise, blood glucose levels, body weight, and abdominal fat mass. The onset is mostly gradual, and symptoms develop slowly over a period of time, with an average of 12 years. It mostly occurs in adults over 30 years old. Symptoms include hyperglycemia, thirst, fatigue, urinary tract infections, nerve disorders, etc. Considering the slow onset, mild symptoms such as repeated infections and weakness are also common. Patients may already have complications at the time of diagnosis, including blindness, retinopathy, heart and kidney diseases, foot lesions, etc.

[0006] Currently, it is believed that T1DM is caused by the autoimmune abnormality mediated by effector T cells, which destroys pancreatic islet β cells, resulting in absolute insulin deficiency. Exogenous administration of insulin is the main and most effective treatment method, but problems such as hypoglycemia, weight gain, exacerbation of insulin resistance, and increased cardiovascular risk occur during long-term insulin therapy. Some non-insulin drugs can assist in lowering blood glucose, and to a certain extent, they can also antagonize the above effects and may have cardiovascular and renal benefits.

[0007] Diabetic ketoacidosis (DKA) is the most common hyperglycemic emergency in patients with diabetes. Without timely and adequate diagnosis and treatment, it is the main cause of death in children with T1DM and one of the main causes of death in adults aged >60 years with severe comorbidities. Potassium loss is obvious in DKA patients, and those with hypokalemia need potassium supplementation.

[0008] As a second-generation aldosterone receptor antagonist, eplerenone is used in the treatment of refractory hypertension such as hypertension caused by primary aldosteronism and heart failure. It has high safety, fewer adverse reactions, and good tolerance; however, there is no report on its use in the treatment of type 1 diabetes. Summary of the Invention

[0009] On the one hand, the present application provides the use of eplerenone in the preparation of a drug for the treatment of type 1 diabetes.

[0010] Type 1 diabetes described in the present application is also known as insulin-dependent diabetes, which is characterized by hyperglycemia caused by absolute insulin deficiency. It is generally believed that the main cause of type 1 diabetes is the destruction of pancreatic islet β cells caused by individual genetic susceptibility or environmental factors, triggering a long-term chronic autoimmune process.

[0011] Furthermore, the rheumatoid arthritis is type 1 diabetes in humans or mice.

[0012] Furthermore, the drug is in oral dosage form or injection dosage form.

[0013] Furthermore, the drug is in oral dosage form.

[0014] Furthermore, the drug further contains pharmaceutically acceptable excipients.

[0015] Furthermore, eplerenone is the only active ingredient in the drug.

[0016] Furthermore, the drug reduces blood glucose levels.

[0017] Furthermore, the drug increases serum insulin levels.

[0018] Furthermore, the drug increases serum C-peptide levels.

[0019] Furthermore, the dosage of the drug is 4 - 100 mg / kg / day.

[0020] The drug dosage in the present application is calculated based on mice. When used in humans or other animals, those skilled in the art can calculate the appropriate dosage level for humans or other animals according to known calculation methods in the art, including but not limited to conversion methods such as body surface area.

[0021] The pharmaceutical composition described in the present invention is in any clinically acceptable dosage form, including various dosage forms for oral and parenteral administration. When used orally, it can be tablets, capsules, oral liquids, syrups, granules, dripping pills, powders, etc.; when used for parenteral administration, it can be new dosage forms such as aqueous injection, powder injection, liposomes, etc.

[0022] According to the different dosage forms, those skilled in the art can select various pharmaceutically acceptable excipients based on the knowledge in the field of pharmacy, including but not limited to fillers, binders, lubricants, disintegrants, solubilizers, surfactants, adsorption carriers, solvents, antioxidants, solubilizers, adsorbents, osmotic pressure regulators, pH regulators, etc.

[0023] The present application discovers that the aldosterone receptor antagonist eplerenone can be used to treat type 1 diabetes, expanding the new clinical indications of eplerenone; it is expected to become an adjuvant drug for immunotherapy of T1DM to play a good hypoglycemic effect; it may be more valuable as a preventive drug for latent autoimmune diabetes in children or adults. In addition, its important potassium-sparing effect is also well-targeted for potassium loss in the body (or intracellular) of DKA patients. Description of the Drawings

[0024] Figure 1 It is a graph of the change in whole blood glucose level.

[0025] Figure 2 It is a graph of the change in serum insulin level.

[0026] Figure 3 It is a graph of the change in serum C-peptide level.

[0027] Figure 4 It is a survival curve graph of each group of mice. Detailed Description of the Invention

[0028] Example 1 Preparation and Administration of Animal Model

[0029] Drug:

[0030] Eplerenone, CAS No.: 107724-20-9, Chemical formula: C 24 H 30 O6, Molecular weight: 414.49, Chemical structure:

[0031]

[0032] Administration method: Oral administration.

[0033] Dissolution method: The solvent is 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline. Weigh an appropriate amount of eplerenone accurately before preparation, and add 10% DMSO → 40% PEG300 → 5% Tween-80 → 45% Saline in sequence to prepare a clear solution. Prepare it once a week and store it at 2℃ - 8℃ after preparation. Restore it to room temperature before administration.

[0034] Usage: The positive drug ruxolitinib (10 mg / kg), the low-dose group of eplerenone (2 mg / kg), the medium-dose group (10 mg / kg), and the high-dose group (50 mg / kg), twice a day for 8 weeks.

[0035] Experimental animal situation:

[0036] Experimental animals: NOD mice, female, 48

[0037] Animal age: 8 weeks old

[0038] Animal source: Jiangsu Jicui Yakang Biotechnology Co., Ltd., production license number: SCXK (Su) 2023-0009, issued by: Jiangsu Provincial Science and Technology Commission.

[0039] Animal feeding: SPF-class animal house, raised in an IVC cage rack, temperature 20 - 26℃, humidity 40 - 70%, 12-hour light-dark cycle; free diet except when fasting is required.

[0040] Experimental animal selection and quantity requirements: NOD mice are an important tool for studying the pathogenesis of type 1 diabetes, and no other non-living animal tests can replace them. On the premise of meeting the research purpose, scientific standards, and regulatory requirements, use as few animals as possible (8).

[0041] Animal welfare: Animal feeding and management are carried out in accordance with the SOP of Suzhou Hekai Biotechnology Co., Ltd., and at the same time refer to the 8th edition of the Guide for the Care and Use of Laboratory Animals (2010) and the Animal Welfare Management Regulations (Public Law 99-198). The method of using animals in this experiment has been approved by the Institutional Animal Care and Use Committee (IACUC) of this institution.

[0042] Model establishment:

[0043] The T1DM model was established by inducing NOD mice with adjuvant injection twice on Day 0 and Day 14.

[0044] Grouping and administration:

[0045] Animals were randomly divided into 6 groups on Day 0 and administered drugs as follows:

[0046] Table 1 Grouping and drug administration of animals

[0047]

[0048] Note: 1 : Ruxolitinib; 2 : p.o., intragastric administration; 3 : BID, twice a day

[0049] Observation indicators:

[0050] Blood glucose, serum insulin, serum C-peptide; survival curve

[0051] Data analysis:

[0052] All data were analyzed by ANOVA variance analysis (GraphPad Prism 8.0 software), * P < 0.05, ** P < 0.01. The obtained data were expressed as and the inter-group differences were analyzed by t-test.

[0053] Experimental results and analysis of Example 2

[0054] Effect of eplerenone on blood glucose:

[0055] Compared with the normal group, the blood glucose data in the model group were significantly increased (P < 0.05); medium-dose eplerenone (10 mg / kg) could significantly reduce the blood glucose level in the early stage of the model (7 - 28 days) (P < 0.05); high-dose eplerenone (50 mg / kg) could significantly reduce the blood glucose level throughout the whole process (P < 0.05, P < 0.01), see Figure 1 .

[0056] Effect of eplerenone on serum insulin level:

[0057] Compared with the normal group, the insulin level in the model group was significantly decreased (P < 0.05); the insulin levels in each dose group of eplerenone were increased compared with the model group, and the increase in the high-dose group was statistically significant (P < 0.05), as shown in Figure 2 .

[0058] Effect of eplerenone on serum C-peptide:

[0059] Compared with the normal group, the C-peptide level in the model group was significantly decreased (P < 0.01); compared with the model group, the C-peptide levels in the low- and medium-dose groups of eplerenone were significantly increased (P < 0.05), as shown in Figure 3 .

[0060] Survival curves of NOD mice in each group

[0061] A total of 3 mice died in the model group (1 mouse died on Day 10, Day 24, and Day 28 respectively), 2 mice died in the low-dose eplerenone group (1 mouse died on Day 10 and Day 28 respectively), and 1 mouse died in the ruxolitinib group (1 mouse died on Day 45). No deaths were observed in the other groups. See Figure 4 Survival curves.

[0062] Two weeks later, the blood glucose in the model group increased significantly and continued to increase until the end of the experiment. Ruxolitinib is a JAK tyrosine kinase inhibitor and is clinically used for the treatment of vitiligo (an autoimmune disease). In this experiment, the increase in blood glucose caused by immune imbalance could be reversed by it, further indicating that the model was successfully established and the modeling rate reached 75%.

[0063] Compared with the model group, 2 mg / kg eplerenone had little hypoglycemic effect, 50 mg / kg eplerenone could significantly reduce blood glucose (P<0.05, comparable to the effect of ruxolitinib), and 10 mg / kg eplerenone had a certain tendency of hypoglycemic effect in the early stage and was between the low and high doses. In addition, at 28 days, the three doses of eplerenone could increase the insulin and C-peptide levels in NOD mice to varying degrees, and some had statistical differences (P<0.05). 3 NOD mice died in the model group, ruxolitinib group, and low-dose eplerenone group respectively, probably due to excessive blood glucose (possibly caused by ketoacidosis), but the blood glucose values could still be included (according to the data before death) to meet the statistical requirements. No deaths were observed in the medium- and high-dose eplerenone groups, indicating that eplerenone has a good protective effect.

[0064] In summary, eplerenone has a good hypoglycemic effect on the type 1 diabetes mouse model, and eplerenone can protect against the damage (death) caused by high glucose. Eplerenone may be a candidate drug for the treatment of type 1 diabetes.

Claims

1. Use of eplerenone in the preparation of a drug for treating type 1 diabetes.

2. The use according to claim 1, wherein the type 1 diabetes is human or mouse type 1 diabetes.

3. The use according to claim 1 or 2, wherein the drug is in an oral dosage form or an injection dosage form.

4. The use according to claim 3, wherein the drug is in an oral dosage form.

5. The use according to any one of claims 1 to 4, wherein the drug further comprises a pharmaceutically acceptable excipient.

6. The use according to any one of claims 1 to 5, wherein eplerenone is the only active ingredient in the drug.

7. The use according to claim 1, wherein the drug lowers blood sugar levels.

8. The use according to claim 1, wherein the drug increases serum insulin levels.

9. The use according to claim 1, wherein the drug increases serum C-peptide levels.

10. The use according to claim 1, wherein the dosage of the drug is 4-100 mg / kg / day.

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