Application of orlistat in preparation of medicine for preventing and treating cisplatin-induced renal injury

Abstract

The invention belongs to the technical field of medicines, and discloses application of orlistat in preparation of a medicine for preventing and treating cisplatin-induced renal injury. The invention discloses application of orlistat or pharmaceutically acceptable salts thereof in prevention and / or treatment of renal injury for the first time. In-vitro cell experiments show that orlistat obviously inhibits toxicity of cis-platinum on renal tubular epithelial cells, DNA damage is inhibited, ROS generation is reduced, endoplasmic reticulum stress is improved, and cell apoptosis is inhibited. Animal experiments show that orlistat can significantly improve the toxicity of cis-platinum to mice, increase the survival rate of the mice, recover the weight of the mice, improve pathological injury of kidneys, inhibit secretion of inflammatory factors, reduce creatinine and urea nitrogen and reverse electrolyte disorder. Particularly, orlistat does not affect the anti-tumor efficacy of cis-platinum at the cellular level and in a tumor-bearing mouse body, and the protection effect of orlistat on kidney injury induced by cis-platinum is superior to that of the unique clinical drug amifostine.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to the use of orlistat in the preparation of a medicament for preventing and treating cisplatin-induced kidney injury. Background Art

[0002] Cisplatin, also known as cis-dichlorodiammineplatinum, began to be used clinically in 1969 and was approved by the FDA in 1978 for the treatment of testicular cancer. Cisplatin is the first platinum-based anti-tumor drug widely used in clinical practice and is also a representative drug of the first generation of platinum-based drugs. Due to its high anti-cancer activity, broad anti-tumor spectrum, and few cross-resistances, since its approval, it has become one of the basic drugs for chemotherapy of various solid tumors. Even at present, while emerging anti-tumor drugs such as targeted drugs and immunotherapy are widely used, cisplatin is still widely used clinically as a first-line basic treatment drug or adjuvant treatment drug for lung cancer, ovarian cancer, osteosarcoma, head and neck tumors, gastric cancer, etc., and is even the preferred combination drug for some advanced malignant tumors. According to statistics, the utilization rate of cisplatin in the chemotherapy of malignant tumors is close to 50%.

[0003] The anti-tumor mechanism of cisplatin is relatively clear: after entering the cell, due to the decrease in chloride ion concentration, cisplatin dissociates and directly binds to DNA to form intra-strand or inter-strand cross-links of DNA, interfering with DNA replication and causing irreversible damage and death of cells. Although cisplatin has good anti-tumor effects, it also has relatively high toxicity, and common and serious toxicities include nephrotoxicity, ototoxicity, electrolyte disorders, etc.

[0004] Although the nephrotoxicity of second-generation and third-generation platinum-based drugs such as carboplatin and oxaliplatin is significantly reduced, their neurotoxicity, myelotoxicity, etc. are significantly increased. Therefore, in clinical applications, the third-generation platinum-based drugs have different clinical indications and application scopes. For more than fifty years, cisplatin remains one of the classic chemotherapy drugs that are irreplaceable for many malignant tumors, but its kidney injury greatly limits its clinical application.

[0005] Nephrotoxicity is the main dose-limiting factor in the clinical use of cisplatin. Among the patients treated with cisplatin clinically, up to 30% of the patients will develop acute kidney injury (AKI). Among the patients who do not meet the AKI criteria after cisplatin treatment, up to 70% of the patients are at risk of developing chronic kidney disease (CKD). Therefore, tumor patients often face the risks of acute and chronic kidney injury when treated with cisplatin.

[0006] Cisplatin is not absorbed orally, so intravenous administration is used clinically. Transporters in the basolateral membrane and luminal membrane of renal tubular cells are responsible for the uptake and efflux of cisplatin. Cisplatin is taken up into renal tubular epithelial cells via organic cation transporter 2 (OCT2), organic anion transporters 1 and 3 (OAT1 / 3), etc. The concentration accumulated in proximal renal tubular epithelial cells can reach 5 times that in the blood. Therefore, even if the blood concentration of cisplatin is non-toxic, it may reach toxic levels in the kidneys. Cisplatin that enters the cells induces DNA damage, oxidative stress, endoplasmic reticulum stress, cell cycle arrest, etc., ultimately leading to inflammation and apoptosis. Depending on the dose and frequency of cisplatin, nephrotoxicity can present as AKI and / or renal interstitial fibrosis. A single high dose of cisplatin causes rapid death of a large number of renal tubular epithelial cells, resulting in acute loss of renal function and the development of AKI; multiple low doses of cisplatin lead to cell cycle arrest of renal tubular epithelial cells, activation of myofibroblasts, and extracellular matrix deposition, resulting in renal interstitial fibrosis and chronic kidney injury.

[0007] In clinical practice, the most commonly used strategies for preventing / ameliorating cisplatin-induced kidney injury are hydration regimens such as normal saline, hypertonic saline, mannitol, and furosemide diuresis. However, due to differences in the definition of nephrotoxicity, the cisplatin dosage, and the patient population in different clinical trials, there is currently no unified standard protocol for cisplatin hydration. In addition, the results of existing clinical trials do not fully support current clinical practice: for example, the renal protective effect of mannitol is not fully supported, and the combination of mannitol and normal saline treatment can increase the incidence of hyponatremia in patients.

[0008] The antioxidant / protective agent amifostine (AMI) is the only drug approved by the US FDA for reducing the nephrotoxicity of ovarian cancer and non-small cell lung cancer. In clinical practice, the recommended drug dose of amifostine is usually 910 mg / m 2 , which is administered by intravenous infusion (completed within 15 minutes) 30 minutes before cisplatin administration. Since amifostine is mainly excreted by the kidneys, the use of high-dose amifostine requires careful consideration of drug concentration in clinical combination for patients with renal insufficiency. At the same time, amifostine itself may cause side effects such as hypotension and nausea. Currently, some clinical trials have attempted low-dose regimens (such as 500 mg / m 2 ) or divided dosing, but the evidence for its efficacy is inconsistent. The effects of amifostine in reversing cisplatin-induced nephrotoxicity vary greatly in different trials, indicating that its protective effect against cisplatin-induced kidney injury is not stable. Therefore, drugs for protecting against cisplatin nephrotoxicity are a major need and urgent requirement in clinical anti-tumor practice.

[0009] Repurposing of existing drugs, also known as drug repositioning or redirection, refers to the discovery of new uses for drugs that are currently being used clinically to treat diseases, as well as candidate drugs that have been or are currently in preclinical or clinical research, through various research methods or practices. Compared with the high investment, long cycle, and high risk of new drug development, repurposing of existing drugs reduces the risk of drug development failure, shortens the development cycle, reduces the development cost, and increases the development success rate, with advantages that are incomparable to "new drugs". In the field of "repurposing of existing drugs" for improving cisplatin-induced damage, there have also been some progress: for example, diphenhydramine has shown certain protective effects against cisplatin nephrotoxicity in cell and animal experiments. Clinical trial results have shown that the proton pump inhibitor pantoprazole can reduce the nephrotoxicity caused by cisplatin in patients with head and neck tumors, but in the study of the application of cisplatin in children and adolescents with osteosarcoma, pantoprazole did not show obvious protective effects against cisplatin nephrotoxicity.

[0010] Orlistat is the first over-the-counter weight loss drug approved by the FDA. It was developed by Hoffmann-La Roche of Switzerland, first launched in New Zealand in 1998, in the United States in 1999, and in China in 2000. Orlistat is a long-acting and potent gastrointestinal lipase inhibitor, and its chemical structure is a derivative of tetrahydrolipstatin. Orlistat inhibits the hydrolysis of approximately 30% of dietary triglycerides by covalently binding to the serine active sites of gastric lipase and pancreatic lipase, reducing the absorption of free fatty acids and monoglycerides. Orlistat has no effect on amylase, trypsin, phospholipase A2, etc., and does not affect the absorption of carbohydrates, proteins, and phospholipids. Orlistat is basically not absorbed after oral administration, and the drug concentration in plasma is very low. After single-dose (maximum dose 800 mg) oral administration, the plasma concentration is <5 ng / mL within 8 hours. 97% of the drug is excreted through feces, and a small amount (<2%) of the original drug and metabolites are excreted from the kidneys. Orlistat has good safety, and there is no drug accumulation in the body after long-term use. The acute toxicity of single-dose oral administration to mice, rats, young rats, and dogs is very low: doses of 1000 mg / kg (dogs), 2000 mg / kg (rats), and 5000 mg / kg (rats, mice) did not show poisoning symptoms. The LD50 of intragastric administration in rats >5 g / kg, and there are no irritating, reproductive toxic, carcinogenic, or mutagenic effects. The side effects of Orlistat are mild, and the main adverse reactions after oral administration by patients are steatorrhea, abdominal discomfort, flatulence, etc.

[0011] The emergence of GLP-1 weight loss drugs such as liraglutide and semaglutide has completely changed the weight loss market. GLP-1 drugs are highly favored, and the market for orlistat has rapidly shrunk due to the "embarrassing" side effect of fat diarrhea. There are many manufacturers and products of orlistat in China, and subsequent competition will surely become even more intense. For pharmaceutical companies, how to respond is an urgent practical issue. Therefore, developing new non-weight loss indications for orlistat helps achieve its secondary development. Summary of the Invention

[0012] In view of the current lack of effective drugs for preventing and treating the renal toxicity of the basic chemotherapy drug cisplatin in cancer treatment, the present invention provides a new use of orlistat, that is, the application of orlistat in the prevention and / or treatment of cisplatin-induced kidney injury.

[0013] The purpose of the first aspect of the present invention is to provide the application of orlistat or a pharmaceutically acceptable salt thereof in the preparation of a product for preventing and / or treating kidney injury.

[0014] The purpose of the second aspect of the present invention is to provide the application of orlistat or a pharmaceutically acceptable salt thereof in the preparation of a drug for reducing cisplatin renal toxicity.

[0015] The purpose of the third aspect of the present invention is to provide a drug.

[0016] In order to achieve the above purposes, the technical solutions adopted by the present invention are as follows:

[0017] The first aspect of the present invention provides the application of orlistat or a pharmaceutically acceptable salt thereof in the preparation of a product for preventing and / or treating kidney injury.

[0018] In some embodiments of the present invention, the kidney injury is drug-induced kidney injury.

[0019] In some embodiments of the present invention, the drug-induced kidney injury is an anti-tumor drug.

[0020] In some embodiments of the present invention, the anti-tumor drug is a platinum-based anti-tumor drug.

[0021] In some embodiments of the present invention, the platinum-based anti-tumor drug is cisplatin.

[0022] In some embodiments of the present invention, the kidney injury includes cisplatin-induced kidney injury.

[0023] In some embodiments of the present invention, the cisplatin-induced kidney injury includes acute kidney injury, chronic kidney injury, and related electrolyte disorders.

[0024] In some embodiments of the present invention, the kidney injury includes acute and chronic kidney injuries caused by a single high dose and / or multiple low doses of cisplatin, as well as related electrolyte disorders.

[0025] In some embodiments of the present invention, the product prevents and / or treats cisplatin-induced kidney injury by improving and reversing cisplatin-induced electrolyte disorders.

[0026] In some embodiments of the present invention, the electrolyte disorder includes at least one of hypomagnesemia and hypocalcemia.

[0027] In some embodiments of the present invention, the product prevents and / or treats cisplatin-induced kidney injury by reducing the expression level of inflammatory factors.

[0028] In some embodiments of the present invention, the inflammatory factors include TNF-α, IL-1β, and IL-6.

[0029] In some embodiments of the present invention, the product prevents and / or treats cisplatin-induced kidney injury by reducing creatinine and blood urea nitrogen.

[0030] In some embodiments of the present invention, the product prevents and / or treats cisplatin-induced kidney injury by reducing the expression of KIM-1, pro-apoptotic protein p53, and cleaved caspase 3.

[0031] In some embodiments of the present invention, the drug prevents and / or treats cisplatin-induced kidney injury by inhibiting cisplatin-induced DNA damage and oxidative stress.

[0032] In some embodiments of the present invention, the drug prevents and / or treats cisplatin-induced kidney injury by inhibiting cisplatin-induced epithelial cell death and apoptosis.

[0033] In some embodiments of the present invention, the drug prevents and / or treats cisplatin-induced kidney injury by inhibiting cisplatin-induced endoplasmic reticulum stress and activation of the MAPKs pathway in epithelial cells.

[0034] The present invention has been proven by experiments that the drug orlistat can significantly inhibit the toxicity of cisplatin-induced rat renal tubular epithelial NRK-52E cells, human proximal renal tubular epithelial cells HK-2 cells, and primary cultured proximal renal tubular epithelial cells (PTEC), reduce the generation of reactive oxygen species (ROS), inhibit DNA damage, inhibit endoplasmic reticulum stress, and inhibit apoptosis. At the same time, orlistat can reduce the mortality of cisplatin-induced normal mice, reduce the levels of blood urea nitrogen and creatinine, and improve the pathological changes of the kidneys; in tumor-bearing mice, orlistat can significantly inhibit the nephrotoxicity of tumor-bearing mice during treatment, reduce the levels of inflammatory factors, and reverse electrolyte disorders. The protective effect of orlistat on cisplatin nephrotoxicity is better than that of amifostine, the only drug approved by the FDA. Orlistat has no effect on the in vitro killing effect of cisplatin on malignant tumor cells, has no effect on the in vivo antitumor activity of cisplatin, and even can enhance the killing effect of cisplatin on tumors.

[0035] In some embodiments of the present invention, the pharmaceutically acceptable salts include at least one of hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, lactate, ascorbate, maleate, tartrate, malate, gluconate, sulfonate, citrate, benzoate, benzenesulfonate, carbonate, mesylate, stearate, nitrate, valerate or succinate.

[0036] In some embodiments of the present invention, the effective dose of orlistat or its pharmaceutically acceptable salt in the product is 10-50 μM.

[0037] The second aspect of the present invention provides the use of orlistat or its pharmaceutically acceptable salt in the preparation of a drug for reducing cisplatin nephrotoxicity.

[0038] In some embodiments of the present invention, the drug reduces cisplatin nephrotoxicity by inhibiting cisplatin-induced DNA damage and oxidative stress (reducing cisplatin-induced ROS).

[0039] In some embodiments of the present invention, the drug reduces cisplatin nephrotoxicity by inhibiting cisplatin-induced epithelial cell death and apoptosis.

[0040] In some embodiments of the present invention, the epithelial cells include renal tubular epithelial cells (such as NRK-52E, HK-2, and PTEC cells).

[0041] In some embodiments of the present invention, the drug inhibits the increase in the expression of p53 and cleaved caspase 3 induced by cisplatin and promotes the expression of the anti-apoptotic protein Bcl-2.

[0042] In some embodiments of the present invention, the drug reduces cisplatin-induced nephrotoxicity by inhibiting the activation of endoplasmic reticulum stress in epithelial cells induced by cisplatin (inhibiting the expression of CHOP and ATF-4 and the phosphorylation of EIF2α) and the MAPKs pathway (inhibiting the phosphorylation of Erk, JNK, and p38 proteins).

[0043] In some embodiments of the present invention, the pharmaceutically acceptable salts include at least one of hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, lactate, ascorbate, maleate, tartrate, malate, gluconate, sulfonate, citrate, benzoate, benzenesulfonate, carbonate, mesylate, stearate, nitrate, valerate, or succinate.

[0044] In some embodiments of the present invention, the effective dose of orlistat or its pharmaceutically acceptable salt in the drug is 10 - 50 μM.

[0045] Orlistat or its pharmaceutically acceptable salt can significantly inhibit the toxicity of cisplatin to renal tubular epithelial cells, inhibit the increase of p53 protein and the decrease of Bcl-2, inhibit the cleavage of Caspase-3, inhibit the activation of MAPKs, endoplasmic reticulum stress, and reduce oxidative stress, and inhibit the apoptosis of renal tubular epithelial cells.

[0046] The third aspect of the present invention provides a drug comprising orlistat or its pharmaceutically acceptable salt and cisplatin.

[0047] Orlistat significantly improves the general condition of mice after cisplatin administration, significantly increases the survival rate of mice, reduces the levels of serum creatinine, urea nitrogen, and inflammatory factors, improves electrolyte disorders, and significantly improves kidney damage. The protective effect of orlistat on cisplatin-induced kidney injury does not affect the in vitro and in vivo antitumor activities of cisplatin, and the protective effect is better than that of amifostine, a drug currently used clinically.

[0048] In some embodiments of the present invention, the drug further comprises a pharmaceutically acceptable excipient, such as at least one of a solvent, a propellant, a solubilizer, a cosolvent, an emulsifier, a coloring agent, a binder, a disintegrant, a filler, a lubricant, a wetting agent, an osmotic pressure regulator, a stabilizer, a glidant, a flavoring agent, a preservative, a suspending agent, a coating material, an aromatic agent, an antiadhesive, a chelating agent, a penetration enhancer, a pH regulator, a buffer, a plasticizer, a surfactant, a foaming agent, a defoaming agent, a thickening agent, a clathrate, a humectant, an absorbent, a diluent, a flocculant and an anti-flocculant, a filter aid, a release retarder, a carrier.

[0049] In some embodiments of the present invention, for the convenience of drug administration, the active ingredient can be processed into a specific dosage form together with any one or several pharmaceutically acceptable excipients. These excipients can be diluents (such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol, and microcrystalline cellulose, etc.), absorbents (such as calcium sulfate, calcium hydrogen phosphate, light magnesium oxide, and calcium carbonate, etc.), wetting agents (such as water and ethanol, etc.), binders (such as hydroxypropyl methylcellulose, povidone, starch paste, and syrup, etc.), disintegrants (such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, effervescent disintegrants, and cross-linked povidone, etc.), lubricants (magnesium stearate, talc, hydrogenated vegetable oil, polyethylene glycol, and colloidal silicon dioxide, etc.), coloring agents (such as titanium dioxide, sunset yellow, methylene blue, and medicinal iron oxide, etc.), coating materials (such as acrylic resin, hydroxypropyl methylcellulose, and povidone, etc.), solvents (such as water for injection, ethanol, propylene glycol, DMSO, and glycerol, etc.), acid-base regulators (such as hydrochloric acid, lactic acid, sodium hydroxide, tartaric acid, and sodium tartrate, etc.), antioxidants (such as sodium sulfite, sodium metabisulfite, and sodium thiosulfate, etc.), bacteriostatic agents (such as phenol, benzyl alcohol, and thimerosal, etc.), or can also be isotonic regulators (such as sodium chloride and glucose, etc.).

[0050] In some embodiments of the present invention, the dosage form of the product includes enteral dosage forms or parenteral dosage forms, preferably parenteral dosage forms.

[0051] In some embodiments of the present invention, the enteral dosage forms include at least one of powders, tablets, granules, capsules, sustained-release agents, solutions, dry suspensions, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0052] In some embodiments of the present invention, the enteral dosage forms include, but are not limited to, enteric-coated tablets, coated tablets, film-coated tablets, sugar-coated tablets, dispersible tablets, sucking tablets, chewable tablets, effervescent tablets, scored tablets, sustained-release controlled-release dosage forms such as sustained-release tablets, sustained-release coated tablets, controlled-release tablets, orally disintegrating tablets, lozenges, etc.

[0053] In some embodiments of the present invention, the parenteral dosage forms include at least one of injection dosage forms, respiratory dosage forms, skin dosage forms, mucosal dosage forms, and cavity dosage forms.

[0054] In some embodiments of the present invention, the injection dosage forms include, but are not limited to, injection solutions, injection suspensions, intravenous drip injection solutions, sterile powders for injection, intravenous injection needles, aqueous injections, injection emulsions, powder injections, injections, sterile powder injections, freeze-dried powder injections, etc.

[0055] The beneficial effects of the present invention are:

[0056] The present invention discloses for the first time the use of orlistat or a pharmaceutically acceptable salt thereof in the prevention and / or treatment of kidney injury. In vitro cell experiments show that orlistat significantly inhibits the toxicity of cisplatin to renal tubular epithelial cells, inhibits DNA damage, reduces ROS production, improves endoplasmic reticulum stress, and inhibits apoptosis. However, orlistat has no inhibitory effect on the killing of tumor cells by cisplatin. In vivo animal experiments show that orlistat can significantly improve the survival rate of mice treated with cisplatin, restore the body weight of mice, improve renal pathological damage, inhibit the secretion of inflammatory factors, reduce creatinine, urea nitrogen, and reverse electrolyte disorders. In particular, orlistat does not affect the anti-tumor efficacy of cisplatin in vivo, and its kidney protection effect is better than that of the only clinically used drug amifostine. This suggests that orlistat or a pharmaceutically acceptable salt thereof is a good drug for the prevention and treatment of cisplatin nephrotoxicity. Orlistat is a drug approved by the FDA and has been clinically used for decades. It has few side effects, high safety, and has the potential to be developed into a drug for protecting against cisplatin nephrotoxicity, with good application prospects and great commercial value. Description of the Drawings

[0057] Figure 1 Orlistat (ORL) inhibits the cytotoxicity of cisplatin (CIS) - induced rat renal tubular epithelial cells NRK - 52E. Among them, A shows that ORL (25 μM) significantly reverses the change in cell morphology induced by cisplatin (left) and MTT assay (right), and the scale bar is 400 μm; B shows propidium iodide (PI) staining, and the scale bar is 400 μm. * represents P < 0.05.

[0058] Figure 2 Orlistat (ORL) inhibits the cytotoxicity of cisplatin - induced human proximal renal tubular epithelial cells HK - 2. Among them, A shows that ORL (25 μM) significantly reverses the change in cell morphology induced by cisplatin (left) and MTT assay (right), and the scale bar is 400 μm; B shows propidium iodide (PI) staining, and the scale bar is 400 μm. * represents P < 0.05.

[0059] Figure 3 Orlistat (ORL) inhibits the cytotoxicity of primary proximal renal tubular epithelial cells (PTEC) extracted from Balb / c mice induced by cisplatin (CIS). Among them, A shows that ORL (25 μM) significantly reverses the change in cell morphology induced by cisplatin (left) and MTT assay (right), and the scale bar is 400 μm; B shows propidium iodide (PI) staining, and the scale bar is 400 μm. * represents P < 0.05.

[0060] Figure 4To investigate the effect of orlistat (ORL) on reversing the expression of apoptosis pathway proteins induced by cisplatin (CIS) in renal tubular epithelial cells. Among them, A, B, and C are the detection of apoptosis pathway proteins P53, Bcl-2, Caspase-3, and cleaved caspase-3 in three cells, namely NRK-52E (A), HK-2 (B), and PTEC (C), by Western Blotting.

[0061] Figure 5 To investigate the effect of orlistat (ORL) on reversing apoptosis of rat renal tubular epithelial cell line NRK-52E induced by cisplatin (CIS). Among them, A is the result of 7-AAD / AnnexinV double staining by flow cytometry; B is the result of TUNEL staining, and the scale bar is 400 μm.

[0062] Figure 6 To investigate the effect of orlistat (ORL) on reversing the generation of reactive oxygen species (ROS) and DNA damage in renal tubular epithelial cells induced by cisplatin (CIS). Among them, A is the result of comet electrophoresis for detecting DNA damage in HK-2 cells, with a magnification of 200; B is the result of flow cytometry for detecting intracellular ROS in NRK-52E cells using the fluorescent probe DCFH 2 -DA. AMI is the clinically approved drug amifostine by the FDA. CON is the blank control group, and H 2 O 2 is the positive control group.

[0063] Figure 7 To investigate the activation of MAPKs pathway and endoplasmic reticulum stress pathway in renal tubular epithelial cell line NRK-52E induced by orlistat (ORL) reversing cisplatin (CIS). Among them, A is the expression of endoplasmic reticulum stress pathway proteins; B is the detection of MAPKs pathway proteins by Western Blotting.

[0064] Figure 8 To investigate that orlistat (ORL) does not affect the killing effect of cisplatin (CIS) on malignant tumor cells. Among them, A - F are the cytotoxic effects of orlistat combined with cisplatin on triple-negative breast cancer 4T1 cells (A), non-small cell lung cancer A549 cells (B), colon cancer HCT116 cells (C), ovarian cancer SKOV3 cells (D), cervical cancer Hela cells (E), and large cell lung cancer H460 cells (F).

[0065] Figure 9Effect of orlistat (ORL) on cisplatin (CIS)-induced renal toxicity in normal mice. Among them, A is a schematic diagram of the animal experiment design process; B is the survival curve of mice; C is the statistical chart of the body weight of mice during the experiment. In each figure, CON is the normal saline group, MODEL is the group given cisplatin alone, ORL-L is the low-dose orlistat combined with cisplatin group, ORL-M is the medium-dose orlistat combined with cisplatin group, ORL-H is the high-dose orlistat combined with cisplatin group, and AMIFOSTINE is the positive drug amifostine combined with cisplatin group.

[0066] Figure 10 Protective effect of orlistat (ORL) on cisplatin (CIS)-induced renal injury in 4T1 tumor-bearing mice. Among them, A is a schematic diagram of the animal experiment design process; B is the statistical chart of the tumor volume of tumor-bearing mice; C is the statistical chart of the body weight of tumor-bearing mice; D is the survival curve between groups of tumor-bearing mice; E is the photo of tumors in each group; F is the photo of kidneys in each group; G is the H&E staining result of kidneys in each group, and the scale bar is 500μm. In the figure, CON is the normal saline group, CIS is the group treated with cisplatin alone, CIS+ORL-L is the low-dose orlistat combined with cisplatin group, CIS+ORL-M is the medium-dose orlistat combined with cisplatin group, CIS+ORL-H is the high-dose orlistat combined with cisplatin group, and CIS+AMI is the positive drug amifostine combined with cisplatin group.

[0067] Figure 11 Protective effect of orlistat (ORL) on cisplatin (CIS)-induced renal injury in 4T1 tumor-bearing mice. Among them, A is the levels of blood urea nitrogen, creatinine, serum magnesium ion, serum calcium ion, serum TNFα, hemoglobin, renal tissue IL-1β and renal tissue IL-6; B is the expression of renal injury biomarker KIM-1, apoptotic protein p53, c-caspase-3, caspase-3 proteins detected by Western Blotting in the kidney tissues of tumor-bearing mice. In the figure, ns represents no significant difference, * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001. Detailed implementation manners

[0068] The content of the present invention will be further described in detail below through specific examples.

[0069] It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention.

[0070] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be described clearly and completely below. For those not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. Reagents or instruments without indicating the manufacturer are all conventional products that can be obtained through commercial purchase.

[0071] The features and performance of the present invention will be further described in detail below in conjunction with the embodiments.

[0072] Orlistat is a long-acting and potent gastrointestinal lipase inhibitor developed by Roche and was first approved for the treatment of obesity in 1998. The chemical structure of Orlistat (Orlistat, ORL) (CAS No.: 96829-58-2) is:

[0073]

[0074] The experimental cell lines used in the embodiments: NRK-52E cells (rat renal tubular epithelial cells), HK-2 cells (human renal cortical proximal tubular epithelial cells), SKOV3 cells, A549 cells, HCT116 cells, HeLa cells, H460 cells, and 4T1 cells were all purchased from the American Type Culture Collection (ATCC); primary mouse renal tubular epithelial cells PTEC were isolated from mouse kidneys and obtained by primary culture.

[0075] Cisplatin used in the embodiments was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; the positive drug amifostine trihydrate was purchased from Shanghai Macklin Biochemical Co., Ltd. Paraformaldehyde, xylene, H&E staining solution, and neutral gum were all purchased from Beijing Solarbio Science & Technology Co., Ltd.

[0076] The antibodies caspase 3 antibody (9662s), p53 antibody (1C12), and p-H2AX (20E3) used in the embodiments were purchased from Cell Signaling (CST); the GAPDH antibody (60004-1-ig) and KIM-1 antibody (30948-1-AP) were purchased from Proteintech.

[0077] Cell culture: NRK-52E cells were routinely cultured with complete medium (DMEM medium containing 5% FBS and 1% double antibody); HK-2 cells were routinely cultured with complete medium (DMEM / F-12 1:1 mixed medium containing 10% FBS and 1% double antibody). During the culture, a constant temperature of 37°C and 5% CO 2 were used.

[0078] Preparation of drug stock solutions: Take orlistat (purchased from Shanghai Macklin Biochemical Co., Ltd., purity > 99%) powder, weigh it, and dissolve it with DMSO solution to prepare a stock solution with a concentration of 25 mM; take cisplatin (purity > 99%) powder, weigh it, and dissolve it with DMSO solution to prepare a stock solution with a concentration of 3 mM. All stock solutions are stored in a -20 °C refrigerator.

[0079] Example 1

[0080] After treating NRK-52E cell line with ORL for 4 hours in advance, cisplatin (15 μM) was given, and after 24 hours, MTT cell viability assay and PI staining were performed on the cells. The specific operations of this example are as follows:

[0081] Take NRK-52E cells in the logarithmic growth phase, seed 5000 cells per well in a 96-well plate, and culture them overnight at 5% CO 2 , 37 °C. After the cells are fully spread and adapted to the seeding environment, remove the complete medium and transfer it to a medium containing 1% FBS. Add 50 μL of a medium solution of orlistat (ORL) at double concentration (0 or 50 μM) with or without, and after 4 hours, add 50 μL of a medium solution of cisplatin at double concentration (0 or 30 μM) with or without. The final concentration of ORL is 25 μM, and the final concentration of cisplatin is 15 μM, and both are given to the cells either in combination or alone. After 24 hours of treatment, cell viability was detected at a wavelength of 570 nm using the conventional MTT method. In addition, after 24 hours of treatment, 20 μL of propidium iodide (PI) (5 mg / mL) was added and incubated, and then observed and photographed under a fluorescence microscope.

[0082] The results are as Figure 1 shown. After treating NRK-52E cells with cisplatin for 24 hours, cell viability decreased from 100% to about 50%. After treating with ORL alone for 24 hours, there was no obvious effect on cell viability. After treating with ORL and cisplatin in combination for 24 hours, cell viability was significantly increased to about 80%; the cell morphology was significantly improved, and the number of cells also increased significantly and normally. PI staining showed that the number of PI-positive cells (stained red) increased after cisplatin treatment, while the number of PI-positive cells in the ORL pretreatment group decreased significantly. This indicates that ORL can significantly protect NRK-52E cells from cisplatin-induced death.

[0083] Example 2

[0084] After treating HK-2 cell line with ORL for 4 hours in advance, cisplatin (30 μM) was given, and after 24 hours, MTT cell viability assay and PI staining were performed on the cells. The specific operations of this example are the same as those in Example 1. The final concentration of ORL is 25 μM, and the final concentration of cisplatin is 30 μM, and both are given to the cells either in combination or alone.

[0085] The results are asFigure 2 As shown in the figure, after HK-2 cells were treated with cisplatin for 24 hours, cell viability decreased from 100% to about 50%. Administration of ORL alone for 24 hours had no obvious effect on cell viability. After ORL and cisplatin were administered jointly for 24 hours, cell viability increased significantly to about 80%; cell morphology was significantly improved, and the number of cells also increased significantly to normal. PI staining showed that the number of PI-positive cells (stained red) increased after cisplatin treatment, while the number of PI-positive cells in the ORL pretreatment group decreased significantly. This indicates that ORL can significantly protect HK-2 cells from cisplatin-induced death.

[0086] Example 3

[0087] After ORL treatment for 4 hours in primary proximal tubular epithelial cells (PTEC cells) extracted from Balb / c mice, 15 μM cisplatin was administered. After 24 hours, MTT cell viability assay and PI staining were performed on the cells. The operation of this example was the same as that of Example 1. The final concentrations were 25 μM ORL and 15 μM cisplatin, administered to the cells either jointly or alone.

[0088] The results were as Figure 3 As shown in the figure, after PTEC cells were treated with cisplatin for 24 hours, cell viability decreased from 100% to about 45%. Administration of ORL alone for 24 hours had no obvious effect on cell viability. After ORL and cisplatin were administered jointly for 24 hours, cell viability increased significantly, cell morphology was significantly improved, and the number of cells also increased significantly. PI staining showed that the number of PI-positive cells (stained red) increased after cisplatin treatment, while the number of PI-positive cells in the ORL pretreatment group decreased significantly. This indicates that ORL can significantly protect PTEC cells from cisplatin-induced death.

[0089] Example 4

[0090] Effect of ORL on the expression of apoptosis-related proteins in cisplatin-induced NRK-52E, HK-2 and PTEC cells. The drug administration method in this example was the same as that in Examples 1-3 above. Western blotting was used to detect the proteins in the apoptosis pathway to investigate the effect of ORL on the apoptosis pathway of renal tubular epithelial cells induced by cisplatin. The specific process was as follows:

[0091] NRK-52E, HK-2 cells in the logarithmic growth phase and primary cultured PTEC cells were respectively cultured in 6-well plates and placed in 5% CO 2Grow overnight at 37°C. After the cells are fully spread and adapted to the plate environment, remove the complete medium and transfer them into the medium containing 1% FBS. After 4 hours with or without ORL at 25 μM, with or without cisplatin (30 μM) is given for another 24 hours. Collect the cells, extract the total protein, and detect the expression of apoptosis-related proteins (caspase 3 protein, cleaved caspase 3 protein, p53 protein, Bcl-2 protein) in each cell by conventional Western blotting.

[0092] The results are as Figure 4 shown. Cisplatin had no significant effect on the total caspase 3 expression in the three types of cells, but significantly increased the cleaved caspase 3, suggesting that cisplatin induced caspase 3-mediated apoptosis; cisplatin significantly increased the expression of the pro-apoptotic protein p53 and significantly decreased the anti-apoptotic protein Bcl-2 in NRK-52E cells; it significantly increased the expression of the pro-apoptotic protein p53 in HK-2 cells. Pretreatment with ORL significantly inhibited the increase in p53 expression and the increase in cleaved caspase 3 induced by cisplatin, and reversed the decrease in Bcl-2 expression. This indicates that under the conditions of the examples, cisplatin significantly induced apoptosis after 24 hours of stimulation, and ORL had a significant protective effect on cisplatin-induced apoptosis in NRK-52E, HK-2 and PTEC cells.

[0093] Example 5

[0094] In the NRK-52E cell line, after treatment with ORL 4 hours in advance, cisplatin at 15 μM was given (the treatment doses of ORL and cisplatin and the experimental procedures were the same as in Example 1). After 24 hours, the cells were collected, incubated with 7-AAD / AnnexinV double probes for staining, and the results of flow cytometry double staining were obtained.

[0095] Use Incucyte to take bright-field and green-field pictures of the TUNEL staining results. TUNEL staining, namely terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling method, the principle of which is based on the cleavage of DNA during apoptosis. When genomic DNA is cleaved, the exposed 3'-OH can be added with fluorescein (FITC)-labeled dUTP (fluorescein-dUTP) under the catalysis of terminal deoxynucleotidyl transferase (TdT), so that it can be detected by fluorescence microscopy or flow cytometry. TUNEL staining is one of the important techniques for measuring apoptosis.

[0096] The results are as Figure 5As shown in the results of 7-AAD / Annexin V flow cytometry double staining, cisplatin induced obvious early apoptosis of cells ( Figure 5 Q3 of A in

[0097] Example 6

[0098] This example investigated the effects of ORL on cisplatin-induced DNA damage and oxidative stress (ROS), as follows:

[0099] NRK-52E cells were treated according to Example 1. After 24 hours, the cells were collected and subjected to comet electrophoresis staining to measure DNA damage, with H 2 O 2 as a control drug for inducing DNA damage. Both the electrophoresis buffer and the lysis buffer were prepared by ourselves. Electrophoresis was carried out in an ice bath at 20 V for 20 minutes in a DNA electrophoresis tank. After electrophoresis, it was washed with deionized water, and 10 μL of PI dye was dripped and photographed under a fluorescence microscope.

[0100] After pre-treating NRK-52E with ORL for 4 hours, 15 μM of cisplatin was given (the treatment method and dosage were the same as in Example 1). After 30 minutes, the cells were collected and incubated with 10 μM of ROS probe DCFH 2 -DA, and the results were measured by flow cytometry. Among them, amifostine (AMI) was used as a positive control drug.

[0101] The results were as Figure 6 shown. The comet electrophoresis results of NRK-52E cells showed that ORL could significantly reduce the DNA tailing phenomenon (comet) caused by cisplatin, indicating that ORL could significantly protect against cisplatin-induced DNA damage. In the flow cytometry results of ROS measurement, cisplatin treatment for half an hour could induce the production of a large amount of ROS in cells (compared with the CON group, the flow peak shifted significantly to the right). ORL could significantly reduce the ROS production induced by cisplatin, and the effect was better than that of amifostine, indicating that ORL could inhibit cisplatin-induced oxidative stress.

[0102] Example 7

[0103] This example was used to investigate the effects of ORL on endoplasmic reticulum stress and MAPKs pathways in cisplatin-induced renal tubular epithelial cells, as follows:

[0104] NRK-52E cells were used, and the administration method and drug dosage were the same as those in Example 1. Western blotting was used to detect the biomarkers of endoplasmic reticulum stress and MAPKs pathway to investigate the effect of ORL on cisplatin-induced endoplasmic reticulum stress and MAPKs.

[0105] The results were as Figure 7 shown. Cisplatin could significantly induce the increased expression of CHOP and ATF-4 proteins in the endoplasmic reticulum stress pathway, and also significantly increase the phosphorylation of EIF2α; the phosphorylation of Erk, JNK, and p38 proteins in the MAPKs pathway was also significantly increased. ORL could significantly inhibit the increased expression of these signaling pathway proteins induced by cisplatin, suggesting that ORL could significantly inhibit the activation of cisplatin-induced endoplasmic reticulum stress and MAPKs pathway.

[0106] Example 8

[0107] This example was used to investigate the effect of ORL on the killing of malignant tumor cells by cisplatin in vitro, and the specific steps were as follows:

[0108] Malignant tumor cells such as SKOV3 cells, A549 cells, HCT116 cells, HeLa cells, H460 cells, and 4T1 cells were used. Each cell was seeded and cultured in a 96-well plate and incubated overnight at 5% CO 2 ₂, 37 °C. After the cells were fully spread and adapted to the seeded environment, the complete medium was removed and the cells were transferred into the medium containing 1% FBS. Each cell was treated with ORL (the final concentration of ORL was between 0 and 200 μM according to different cell types) 4 hours in advance and then given cisplatin (15 μM). The treatment method was the same as that in Example 1. After 24 hours, MTT cell viability assay was performed on the cells.

[0109] The results were as Figure 8 shown. Administering ORL alone did not promote the proliferation of each malignant tumor cell, and on A549 cells, HCT116 cells, and H460 cells, ORL could also inhibit the proliferation of malignant tumor cells in a concentration-dependent manner. In the experimental group co-administered with ORL and cisplatin, it was completely different from that in renal tubular epithelial cells: ORL at different concentrations not only had no obvious protective effect on cisplatin-induced death of malignant tumor cells, but could significantly enhance the killing activity of cisplatin at slightly higher concentrations. This showed that ORL did not affect the anti-tumor drug activity of cisplatin in vitro.

[0110] Example 9

[0111] This example used animal experiments to investigate the effect of ORL on cisplatin-induced renal injury in normal mice. Normal mice were selected to investigate whether ORL would inhibit the renal injury effect of cisplatin itself at the animal level, and the specific steps were as follows:

[0112] SPF-grade Balb / c mice aged 6 - 7 weeks (25 g ± 3 g, sourced from the Faculty of Health Sciences, University of Macau) were acclimatized to the environment for one week and then caged and grouped as DAY 0. They were divided into a CON group, a MODEL group, a low-dose ORL group (ORL-L group), a medium-dose ORL group (ORL-M group), a high-dose ORL group (ORL-H group), and a positive drug AMIFOSTINE group (i.e., the positive drug amifostine), with 6 mice in each group. The CON group was intraperitoneally injected with an equal volume of normal saline only; the MODEL group was intraperitoneally injected with cisplatin (15 mg / kg) once; the low, medium, and high-dose ORL groups, on the basis of a single injection of cisplatin, were intraperitoneally injected with ORL once a day, with doses of 1 mg / kg, 3 mg / kg, and 10 mg / kg respectively. The cisplatin working solution was intraperitoneally injected at 15 mg / kg, only once, and the orlistat working solution was injected daily; the AMI FOSTINE group was administered in the same way as the ORL group, that is, on the basis of a single injection of cisplatin, AMIFOSTINE was intraperitoneally injected once a day at a dose of 200 mg / kg. Starting from DAY 1, the body weights of the mice were recorded daily. On the 10th day, the mice were observed quietly without being given drugs. After recording the body weights on the 15th day, euthanasia was performed.

[0113] The results are as Figure 9 shown. Compared with the CON group, the body weights of the mice in the other groups decreased significantly. The body weights of the mice in the positive drug AMI or ORL groups were higher than those in the MODEL group in the later stage. At the same time, the body weight of the ORL-H group began to stabilize and showed an upward trend on the 12th day. All the mice in the MODEL group died on the 7th day, all the mice in the ORL-L group and the AMIFOSTINE group died on the 12th day, and all the mice in the ORL-M group died on the 13th day; while in the ORL-H group, except for one mouse that died on the 10th day, the rest of the mice survived until the end of the experiment, and at the end of the experiment, the body weights of the mice in the ORL-H group showed an obvious upward trend. These results suggest that ORL can significantly counteract the nephrotoxicity of a single high dose of cisplatin, significantly extend the survival curve of mice, and inhibit the decrease in the body weight of mice, and its effect is better than that of the positive drug amifostine.

[0114] Example 10

[0115] This example uses animal experiments to investigate the effect of ORL on cisplatin-induced renal injury in tumor-bearing mice.

[0116] Since cisplatin is a basic chemotherapy drug and is widely used in the treatment of various malignancies. One of the necessary prerequisites for the application of ORL to inhibit and protect its renal injury is that ORL does not weaken the anti-tumor efficacy of cisplatin. Therefore, in this example, a xenograft tumor model of triple-negative breast cancer 4T1 cells was selected to investigate whether ORL would affect the anti-tumor efficacy of cisplatin. Clinically, when cisplatin chemotherapy is used, it is generally carried out for 2 - 3 weeks, with one week as a cycle, and cisplatin is administered 2 - 3 times within each cycle. This example adopted a similar protocol to the clinical use of cisplatin, and the specific protocol is as follows:

[0117] After 6 - 7-week-old SPF-grade Balb / c mice (25g ± 3g, sourced from Zhuhai BestBio Technology Co., Ltd.) were adapted to the environment for one week, all mice were subcutaneously injected with 1×10 6 4T1 cells one week in advance for modeling. After the tumors were successfully modeled, the mice were randomly divided into cages and grouped, and recorded as DAY 1. The tumor-bearing mice were grouped into a CON group, a cisplatin group (CIS group), low, medium, and high-dose orlistat groups (CIS+ORL-L, CIS+ORL-M, CIS+ORL-H), and an amifostine group (CIS+AMI). The CON group was intraperitoneally injected with an equal volume of normal saline once a day; the CIS group was intraperitoneally injected with cisplatin at 5mg / kg every 3 days, 2 times a week; the low, medium, and high-dose orlistat groups were intraperitoneally injected with ORL at 1mg / kg, 3mg / kg, and 10mg / kg respectively once a day on the basis of cisplatin treatment; the amifostine group was intraperitoneally injected with amifostine at 200mg / kg once a day on the basis of cisplatin treatment.

[0118] Starting from DAY 1, the body weight and tumor volume of the mice were recorded every day. After recording the body weight and tumor volume on the 25th day, euthanasia was performed for sample collection. Blood was taken, and the tumor tissues of the mice were quickly removed and weighed, and the kidneys were washed in physiological saline, observed and photographed. The right kidney was stripped of the surface adipose tissue and mucosa and fixed in 4% paraformaldehyde for histopathological examination, and the remaining parts were labeled and placed in a -80°C refrigerator for other related detections.

[0119] Kidney tissue was excised, total protein was extracted, protein quantification was performed by the BCA method, and the expressions of KIM-1, p53, and cleaved caspase3 in the kidney tissue were detected by conventional Western Blotting.

[0120] Histopathological examination: After the kidney tissue was fixed in 4% paraformaldehyde, it was routinely paraffin-embedded and cut into 4μm-thick sections for conventional H&E staining. Observation, photography, and analysis were performed under a light microscope.

[0121] Renal function detection: The levels of blood creatinine (BUN) and blood urea nitrogen (CREA) were detected using a dry biochemical analyzer (Jiangsu Kangshang Biomedical Technology Co., Ltd.).

[0122] Inflammatory response detection: A commercial ELISA kit (Xinbosheng Biotechnology Co., Ltd.) was used to detect the levels of TNF-α in serum and IL-1β and IL-6 in the kidneys.

[0123] Electrolyte disorder detection: A commercial kit (Sangon Biotech Co., Ltd.) was used to detect the levels of electrolyte magnesium ions and calcium ions in serum.

[0124] The results were as Figures 10 to 11 shown. Compared with the CON group, the tumor volumes in the remaining groups decreased significantly. The positive drug amifostine or ORL combined with cisplatin had no effect on the tumor volume and did not weaken the anti-tumor activity of cisplatin. During the entire experiment, one mouse in the CIS group died on the 16th day, one mouse in the group of positive drug amifostine combined with cisplatin died on the 12th and 18th days respectively, while no mouse in the CON and ORL groups died. Renal function tests showed that cisplatin significantly induced an increase in BUN and CREA. ORL and amifostine significantly reversed the effect of cisplatin, and the reversal effect of ORL was similar to that of amifostine. The test of inflammatory factor levels showed that ORL could dose-dependently reduce the levels of TNF-α, IL-1β and IL-6. The detection of electrolyte levels showed that cisplatin significantly induced hypomagnesemia and hypocalcemia, indicating electrolyte disorders, which could be almost completely reversed by ORL.

[0125] H&E staining is also known as hematoxylin & eosin staining. After staining, the cell nucleus appears blue-purple and the cytoplasm appears pink. The H&E staining results of the kidneys showed that compared with the CON group, the kidneys of mice in the CIS group had obvious damage, vacuoles or swelling, and both ORL and amifostine administration could significantly improve the above pathological changes. The Western Blotting results of renal tissues showed that cisplatin induced a significant increase in the expression of the biomarker KIM-1 for renal injury and an increase in the expression of the pro-apoptotic proteins p53 and cleaved caspase 3. In each group treated with ORL, these changes were reversed to varying degrees, and the reversal effect of ORL was significantly better than that of the positive drug amifostine. These results suggest that while ORL improves cisplatin-induced renal injury in tumor-bearing mice, it does not affect the anti-tumor efficacy of cisplatin, and the effect is better than that of the positive control drug amifostine in some indicators.

[0126] It is particularly worth mentioning that no kidney stones or calcium oxalate crystals were found in the kidneys of the ORL administered by intraperitoneal injection in this example. There are research reports that oral ORL inhibits fat decomposition, resulting in the binding of fat and calcium in the gastrointestinal tract, competitively inhibiting the binding of oxalic acid and calcium, causing excessive oxalic acid to be absorbed by the intestine, and increasing the excretion of oxalic acid by the kidneys. Eventually, supersaturation of oxalic acid occurs in the renal tubules, forming calcium oxalate crystals or kidney stones. Therefore, this suggests that the side effects of kidney stones or calcium oxalate crystals caused by ORL may be related to the administration method. In this example, ORL was administered by intraperitoneal injection, avoiding the inhibition of local fat decomposition in the gastrointestinal tract, not affecting the binding of oxalic acid and calcium in the gastrointestinal tract, not promoting the increase of oxalic acid absorption in the gastrointestinal tract and the increase of oxalic acid excretion by the kidneys. Therefore, intravenous administration of ORL will avoid the potential renal damage caused by oral ORL.

[0127] From the above pharmacological experimental results, it can be seen that ORL has a significant effect of inhibiting cisplatin nephrotoxicity and protecting the kidneys, and does not affect the anti-tumor efficacy of cisplatin in vivo and in vitro, showing the application prospect of being developed into a drug for improving cisplatin-induced kidney injury.

[0128] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the scope of knowledge possessed by those of ordinary skill in the art. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. Use of orlistat or a pharmaceutically acceptable salt thereof in the preparation of a product for preventing and / or treating renal injury.

2. The use according to claim 1, characterized in that: The renal injury is drug-induced renal injury; Preferably, the drug-induced renal injury is an anti-tumor drug; Preferably, the anti-tumor drug is a platinum anti-tumor drug; Preferably, the platinum anti-tumor drug is cisplatin.

3. The use according to claim 1, characterized in that: The renal injury includes renal injury induced by cisplatin.

4. The use according to claim 3, characterized in that: The cisplatin-induced renal injury includes acute renal injury, chronic renal injury and electrolyte disturbances associated therewith.

5. The use according to claim 4, characterized in that: The product prevents and / or treats cisplatin-induced renal injury by improving and reversing electrolyte imbalance induced by cisplatin; and / or, the product prevents and / or treats cisplatin-induced renal injury by reducing the expression level of inflammatory factors; and / or, the product prevents and / or treats cisplatin-induced renal injury by reducing creatinine and urea nitrogen.

6. The use according to claim 5, characterized in that: The electrolyte disorder includes at least one of hypomagnesemia and hypocalcemia; and / or the inflammatory factors include TNF-α, IL-1β and IL-6.

7. Use of orlistat or a pharmaceutically acceptable salt thereof in the preparation of a drug for reducing cisplatin nephrotoxicity.

8. The use according to claim 7, characterized in that: The drug reduces the nephrotoxicity of cisplatin by inhibiting cisplatin-induced DNA damage and oxidative stress; and / or, the drug reduces the nephrotoxicity of cisplatin by inhibiting cisplatin-induced epithelial cell death and apoptosis; and / or, the drug reduces the nephrotoxicity of cisplatin by inhibiting cisplatin-induced epithelial cell endoplasmic reticulum stress and activation of the MAPK pathway.

9. The use according to any one of claims 1 to 8, characterized in that: The pharmaceutically acceptable salt includes at least one of hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, lactate, ascorbate, maleate, tartrate, malate, gluconate, sulfonate, citrate, benzoate, benzenesulfonate, carbonate, methanesulfonate, stearate, nitrate, valerate or succinate.

10. A medicament comprising orlistat or a pharmaceutically acceptable salt thereof and cisplatin; Preferably, the drug further comprises a pharmaceutically acceptable excipient.

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