Application of IL17 inhibitor in preparation of medicine for treating diabetic acute kidney injury

Abstract

The invention discloses an application of an IL-17 inhibitor (SGC-CBP30) in preparation of a medicine for treating diabetic acute kidney injury. Research results show that the SGC-CBP30 can significantly inhibit activation of an IL17 pathway in a diabetic ischemia reperfusion kidney and related inflammatory injury, and also can significantly down-regulate expression of M1 type macrophage related inflammatory factors IL6 and IL1B and Th17 cell key transcription factors PSTAT3, RORC and IL17A in kidney tissues, and relieve renal tubular injury and renal fibrosis. The application of the IL17 inhibitor in preparation of the medicine for treating the diabetic acute kidney injury is expanded, the research result provides a new theoretical basis for basic research of the diabetic acute kidney injury, and a new way is expected to be developed for prevention and treatment of the diabetic acute kidney injury.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical treatment, and relates to the application of an IL17 inhibitor in the preparation of a drug for treating diabetic acute kidney injury. Background Art

[0002] Type 2 diabetes mellitus (T2DM) is one of the chronic diseases that currently poses a serious threat to global human health. It affects 463 million people worldwide, and this number may increase to 700 million by 2045. Long-term hyperglycemia, insulin resistance, and metabolic disorders can cause various acute and chronic organ damages in the body. The kidney is one of the most commonly affected organs in diabetes. About 40% of patients with type 2 diabetes may experience proteinuria and / or renal impairment. Diabetes is also one of the main causes of chronic kidney disease and end-stage renal disease, which has brought a heavy burden to families and society.

[0003] As our understanding of the long-term risks of diabetes continues to evolve, so does our understanding of the myriad challenges that diabetes and its complications can present. Acute kidney injury (AKI) is a complication of diabetes that requires attention and is a public health problem that causes a high socioeconomic burden worldwide. The main feature of AKI is a sudden decline in renal excretion function, manifested by an increase in serum creatinine levels and a decrease in urine output. In recent years, the etiology of AKI has changed: increasing evidence suggests that DM is an independent risk factor for the development of AKI, while another view is that AKI is a true complication of DM. Clinically, a high proportion of children with type 1 diabetes (T1DM) hospitalized for diabetic ketoacidosis (DKA) will develop AKI; compared with non-diabetic patients, patients with type 2 diabetes have an increased risk of AKI and mortality. The pathophysiology of AKI is multifactorial and complex. Renal ischemia-reperfusion injury (IRI) is a common pathophysiological phenomenon of AKI and is also the main cause of AKI. It has a high mortality rate and there is still a lack of effective treatment drugs.

[0004] The IL-17 inhibitor (SGC-CBP30) is a highly selective and effective inhibitor of the CBP / p300 bromodomain, with Kd values ​​of 21 and 32 nM for CBP and p300, respectively. It strongly reduces the secretion of IL17A in Th17 cells and has anti-inflammatory effects. In ankylosing spondylitis and psoriatic arthritis, SGC-CBP30 can inhibit the secretion of IL17A by Th17 cells. In vitro studies also showed that SGC-CBP30 treatment can slightly alleviate NSC-125066-induced alveolar bronchial fibrosis. There are also studies showing that the inhibitor SGC-CBP30 can alleviate intestinal radiation damage. However, there are no reports in the literature on the use of IL17 inhibitors (SGC-CBP30) in the preparation of drugs for the treatment of diabetic acute kidney injury. Summary of the invention

[0005] The present invention discloses for the first time the use of IL17 inhibitors in the preparation of drugs for treating diabetic acute kidney injury. Experimental results show that IL17 inhibitor SGC-CBP30 can significantly alleviate diabetic acute kidney injury, providing a new option for the preparation of drugs for diabetic acute kidney injury.

[0006] The IL-17 inhibitor (SGC-CBP30) for treating acute kidney injury in diabetes disclosed in the present invention can be prepared into various pharmaceutical compositions with one or more pharmaceutically acceptable carriers, excipients or diluents, including various solid oral preparations, liquid oral preparations, etc.

[0007] The IL-17 inhibitor (SGC-CBP30) for treating acute kidney injury in diabetes of the present invention is effective within a fairly wide dosage range. For example, the daily dosage may be in the range of approximately 0.1 mg-1000 mg per kilogram of body weight. In the treatment of adults, the dosage range is preferably 1 mg / kg-10 mg / kg, administered once or several times. The actual dosage of the IL-17 inhibitor (SGC-CBP30) for treating acute kidney injury in diabetes of the present invention should be determined by the doctor based on relevant circumstances, including the physical condition, age, weight, individual response of the patient to the drug, severity of the patient's symptoms, etc., and therefore the above dosage range does not limit the scope of the present invention in any way.

[0008] When administered orally, the composition can be formulated into tablets, dispersible tablets, sugar-coated tablets, granules, dry powders, solutions or capsules. Lactose or starch can be used as carriers for preparing oral pharmaceutical compositions, and gelatin, sodium carboxymethylcellulose, methylcellulose, polyvinyl pyrrolidone, etc. are suitable binders. Starch or microcrystalline cellulose can be selected as disintegrants, often talcum powder, colloidal silica, glyceryl stearate, calcium or magnesium stearate, polyethylene glycol-4000, polyethylene glycol-6000, sodium metabisulfite, etc.; as suitable anti-adhesives and lubricants. For example, tablets can be prepared by pressing wet granules. The active ingredient is mixed with a carrier and optionally with a portion of a disintegrant additive, and the mixture is granulated with an aqueous solution, an alcoholic or an aqueous alcoholic solution of a binder in a suitable device, and the dried granules are then added with other disintegrants, lubricants and anti-adhesives to tablet the mixture.

[0009] A more detailed introduction to IL17 inhibitors in the treatment of diabetic acute kidney injury is as follows: Methods: This study used a high-fat diet combined with intraperitoneal injection of streptozotocin (STZ) to establish a type 2 diabetes model, and bilateral renal pedicle clamping and reperfusion were used to prepare the model. In vitro, a cell injury model was established by high glucose combined with hypoxia and reoxygenation. SGC-CBP30 was used in vivo and in vitro to observe its effects on renal immune response, tubular injury and renal fibrosis in diabetic ischemia-reperfusion injury.

[0010] Steps: Animal Experiments Animal model establishment and grouping The type 2 diabetes model was established by feeding with a 45% high-fat diet for 8 weeks and intraperitoneal injection of 60 mg / kg / d STZ solution for 5 consecutive days. The non-diabetic group was injected with an equal dose of 0.1 mmol / L sodium citrate buffer as a control. After the model was successfully established (random blood glucose > 16.7 mmol / L one week after STZ injection), the changes in body weight and random blood glucose were recorded weekly.

[0011] According to the weight of the mice, 60 mg / kg 2% pentobarbital was injected intraperitoneally for anesthesia. After the anesthesia took effect, the mice were fixed on the operating table and the back was depilated and disinfected. The skin was cut open near the side of the spine to separate the kidney, and the perirenal fat tissue and renal pedicle were bluntly separated. After the bilateral renal pedicles were clamped by arterial clamps, the time was counted for 25 minutes respectively. The interval between the clamping of the bilateral renal pedicles did not exceed 1 minute. The kidney changed from bright red to purple-red, indicating successful blockage. After the arterial clamp was released, the kidney changed from purple-red to bright red, indicating successful renal reperfusion. After the kidney was repositioned, double-layer suture was given and observed in a single cage. After the operation, it was kept warm and supplemented with an appropriate amount of normal saline. The medication group needed to be injected with 10 mg / kg SGC-CBP30 intraperitoneally 8 hours after reperfusion. The renal pedicles of mice in the sham operation group were not clamped, and other surgical operations were the same as before. The status of the mice was closely observed after surgery.

[0012] Detection of mouse renal function After blood was collected from the eyeball, the mice were killed, and the venous blood was centrifuged and the supernatant was drawn out to measure blood urea nitrogen and creatinine using an automatic biochemical analyzer to calculate the creatinine clearance rate.

[0013] 3. Observation of M1 macrophage and Th17 cell infiltration in mouse kidneys Western blot was used to detect the expression levels of the main inflammatory factors (IL6, IL1B) secreted by M1 macrophages and the main transcription factors (PSTAT3, RORC) of Th17 cells.

[0014] IV. Detection of IL17 pathway activation, renal tubular injury and fibrosis in mouse kidneys Western blot was used to detect the activation of key proteins and effector factors of the IL17 signaling pathway in renal tissue, as well as the expression of renal tubular injury-related factors and renal fibrosis-related proteins.

[0015] Peripheral blood mononuclear cells were co-cultured with proximal tubular epithelial cells, and 20 μmol SGC-CBP30 was added to the drug group at 30 minutes after reoxygenation. Western blot was used to detect the activation of key proteins and effector factors of the IL17 signaling pathway in proximal tubular epithelial cells, as well as the expression of tubular injury-related factors and renal fibrosis-related proteins.

[0016] Results: The results showed that SGC-CBP30 could significantly inhibit the activation of IL17 pathway and related inflammatory damage in diabetic ischemia-reperfusion kidneys, and also significantly down-regulate the expression of M1 macrophage-related inflammatory factors IL6 and IL1B, Th17 cell key transcription factors PSTAT3, RORC and IL17A in renal tissue, thereby alleviating renal tubular damage and renal fibrosis.

[0017] Conclusion: In addition to significantly inhibiting the activation of the downstream IL17 pathway, tubular injury and renal fibrosis, SGC-CBP30 also has a certain effect on macrophages and Th17 cells. Through this study, the application of IL17 inhibitors in the preparation of drugs for the treatment of diabetic acute kidney injury has been expanded. The research results provide a new theoretical basis for the basic research of diabetic acute kidney injury and are expected to open up new ways for its prevention and treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 SGC-CBP30 can alleviate renal function damage in diabetic mice with renal ischemia-reperfusion injury; A: serum creatinine; B: blood urea nitrogen; *P < 0.05, **P < 0.01, ***P < 0.001 Figure 2 SGC-CBP30 significantly reduced the Th17 immune response and inflammatory damage in the kidneys of diabetic ischemia-reperfusion injury; AB: Western blot was used to detect the protein expression levels of IL6, IL1B, RORC, PSTAT3, and IL17 in renal tissues and their quantitative analysis; CD: Western blot was used to detect the protein expression levels of IL17RA, PP38 / P38, PERK / ERK, and PP65 / P65 in renal tissues and their quantitative analysis; EF: Western blot was used to detect the protein expression levels of MMP3, TNFA, COX2, CCL3, and CCL2 in renal tissues and their quantitative analysis; GH: Western blot was used to detect the protein expression levels of ECA, VIMENTIN, SMA, TGFB, AKLOTHO, and EDN1 in renal tissues and their quantitative analysis; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; Figure 3 SGC-CBP30 significantly reduced the activation of the IL17 signaling pathway and inflammatory damage in proximal tubular epithelial cells; E: Western blot was used to detect the protein expression level of IL17RA and the phosphorylation activation of P38, ERK, and P65 in proximal tubular epithelial cells co-cultured with peripheral blood mononuclear cells under different intervention conditions, and their quantitative analysis; FR: Western blot was used to detect the protein expression levels of IL17 signaling pathway effectors MMP3, TNFA, COX2, CCL3, CCL2, and fibrosis-related proteins ECA, VIMENTIN, SMA, and TGFB, as well as tubular injury-related factors AKLOTHO and EDN1 in proximal tubular epithelial cells co-cultured with peripheral blood mononuclear cells under different intervention conditions, and their quantitative analysis; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. DETAILED DESCRIPTION

[0019] The present invention is described below by specific embodiments. Unless otherwise specified, the technical means used in the present invention are methods known to those skilled in the art. In addition, the embodiments should be understood as illustrative rather than limiting the scope of the present invention, and the essence and scope of the present invention are limited only by the claims. For those skilled in the art, various changes or modifications to the material components and dosages in these embodiments, without departing from the essence and scope of the present invention, also belong to the protection scope of the present invention. The raw materials and reagents used in the present invention are all commercially available. Among them, SGC-CBP30 was purchased from TargetMol, USA. Example

[0020] Preparation of tablets containing 100 mg SGC-CBP30 per tablet: mg / tablet SGC-CBP30 100 Lactose 50 Microcrystalline Cellulose 80 Starch 50 Hydroxymethylcellulose 40 Magnesium Stearate 5 The active ingredients, lactose, starch and microcrystalline cellulose are passed through a 100-mesh sieve and mixed thoroughly. A 2% hydroxymethylcellulose aqueous solution is added to the mixed powder and mixed. The mixture is passed through a 20-mesh sieve to prepare a soft material. The wet granules are dried at 45-55°C. Sodium carboxymethyl starch and magnesium stearate are added to the dry granules and tableted. Example

[0021] Capsules containing 100 mg SGC-CBP30 per capsule were prepared as follows: Dosage / capsule weight concentration (%) SGC-CBP30 100mg 30.0 Polyoxyethylene dehydrated sorbitan 0.05mg 0.02 Sugar alcohol monooleate Starch 250mg 69.98 Total 350.05 mg 100.00.

Claims

1. Application of IL17 inhibitors in the preparation of drugs for the treatment of diabetic acute kidney injury.

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