Use of nitrofurazone or pharmaceutically acceptable derivatives thereof and pharmaceutical compositions

Abstract

The application provides a nitrofurazone or a pharmaceutically acceptable derivative thereof, and belongs to the technical field of biological medicines. The application comprises the use of nitrofurazone in the preparation of a drug for reducing food intake and / or inhibiting appetite. The nitrofurazone can be independent of blood sugar regulation, independent of the gastrointestinal tract, and does not cause side effects such as nausea and vomiting; the target of the nitrofurazone is not located in the peripheral nervous system, but acts on the central nervous system. The nitrofurazone can significantly reduce the intake of ordinary food and high-fat food of mice in the form of activating Glp1-r, inhibit the appetite of the mice, and thus significantly reduce the body weight of obese mice. Therefore, the nitrofurazone has the potential to treat obesity and can be applied to the preparation of a drug for treating obesity.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, in particular to the application and pharmaceutical composition of furazolidone or its pharmaceutically acceptable derivatives. BACKGROUND

[0002] Obesity is a chronic metabolic disease caused by multiple factors, mainly manifested as excessive accumulation of fat in the body, leading to abnormal increase in body weight (BMI≥30).

[0003] At present, the treatment of obesity mainly relies on lifestyle intervention, drug treatment and metabolic surgery. Lifestyle intervention (such as diet control, exercise and behavior therapy) is the basic means, but the long-term compliance is poor, and the effect on moderate and severe obesity patients is limited. In terms of drug treatment, existing drugs such as glucagon-like peptide-1 (Glp-1) receptor agonists (such as semaglutide), pancreatic lipase inhibitors (such as orlistat) can reduce body weight, but there are side effects (such as hypoglycemia, gastrointestinal reactions, risk of pancreatitis) and weight rebound after drug withdrawal. Metabolic surgery (such as gastric bypass surgery) can significantly reduce weight and improve metabolic abnormalities, but the surgery is high-risk, expensive, and may cause malnutrition or postoperative complications, and is only suitable for a small number of patients. These limitations highlight the urgent need to develop new anti-obesity drugs, and the ideal new drug should have better safety, more significant weight loss effect and multiple benefits in improving metabolic indicators. SUMMARY

[0004] Therefore, in order to at least partially solve at least one of the above-mentioned technical problems, the present application provides the application of furazolidone or its pharmaceutically acceptable derivatives, which includes the application of furazolidone in the preparation of a drug for reducing food intake and / or suppressing appetite.

[0005] According to an embodiment of one aspect of the present application, a pharmaceutical composition for reducing food intake and / or treating obesity is provided, which comprises furazolidone or its pharmaceutically acceptable derivatives as an active ingredient.

[0006] According to the embodiment of the present application, furazolidone can be independent of blood glucose regulation, independent of the gastrointestinal tract, and does not cause side effects such as nausea and vomiting; its target is not located in the peripheral nervous system, but acts on the central nervous system. Furazolidone can significantly reduce the intake of ordinary food and high-fat food in mice in the form of activating Glp1-r, suppress the appetite of mice, and thus significantly reduce the body weight of obese mice. Therefore, furazolidone has the potential to treat obesity and can be used in the preparation of a drug for treating obesity. BRIEF DESCRIPTION OF DRAWINGS

[0007] Figure 1Figure for change of ordinary food intake after intraperitoneal injection of nitrofurazone to mice in Example 1 of the present application;

[0008] Figure 2 Figure for change of ordinary food intake after intraperitoneal injection of nitrofurazone to mice after 12 hours of fasting in Example 1 of the present application;

[0009] Figure 3 Figure for change of relative blood glucose after intraperitoneal injection of nitrofurazone to mice after 12 hours of fasting in Example 1 of the present application;

[0010] Figure 4 Comparison figure for overnight high-fat food intake after intraperitoneal injection of nitrofurazone to mice in Example 1 of the present application;

[0011] Figure 5 Figure for change of body weight of obese mice after intraperitoneal injection of nitrofurazone in Example 1 of the present application;

[0012] Figure 6 Statistical figure for ordinary food intake after intraperitoneal injection of nitrofurazone to mice after removal of enterogastric vagus nerve in Example 2 of the present application;

[0013] Figure 7 Statistical figure for ordinary food intake after intraperitoneal injection of nitrofurazone to mice after removal of enterogastric vagus nerve after 12 hours of fasting in Example 2 of the present application;

[0014] Figure 8 Statistical figure for number of times of retching within 3 hours after intraperitoneal injection of nitrofurazone to mice in Example 2 of the present application;

[0015] Figure 9 Statistical figure for retching latency within 3 hours after intraperitoneal injection of nitrofurazone to mice in Example 2 of the present application;

[0016] Figure 10 Statistical figure for number of times of defecation within 3 hours after intraperitoneal injection of nitrofurazone to mice in Example 2 of the present application;

[0017] Figure 11 Figure for detection result of marker protein (c-Fos protein) of hypothalamic paraventricular nucleus (PVN) brain area neurons after intraperitoneal injection of nitrofurazone to obese Glp1r-cre::Ai14 mice after high-fat modeling in Example 3 of the present application;

[0018] Figure 12 Statistical figure for c-Fos protein of PVN brain area after intraperitoneal injection of nitrofurazone to Glp1r-cre::Ai14 mice in Example 3 of the present application, wherein A is a statistical figure for paraventricular nucleus, and B is a statistical figure for Glp-1 receptor. DETAILED DESCRIPTION

[0019] Embodiments of the present application will be described herein below with reference to the accompanying drawings. It is to be understood, however, that such description is merely exemplary and is intended to provide a thorough and complete disclosure of embodiments of the present application, as copies can be filed. In the following detailed description of embodiments of the present application, numerous specific details are set forth in order to provide a thorough understanding of embodiments of the present application. However, it will be apparent to one skilled in the art that one or more embodiments of the present application can be practiced without these specific details. In other instances, well-known structures and methods have not been described in detail in order to avoid unnecessarily obscuring embodiments of the present application.

[0020] The terms used herein are merely for the purpose of describing particular embodiments and are not intended to limit the present application. The term "comprising" used herein indicates the presence of a feature, step, operation, but does not exclude the presence or addition of one or more other features.

[0021] In the present application, the term "pharmaceutically acceptable" means a compound, material, composition, and / or dosage form that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0022] The term "treatment" means that after the onset of a disease, a subject is contacted (e.g., administered) with a drug, composition, etc. based on the present application, thereby reducing the symptoms of the disease compared to not being contacted, and does not mean that the symptoms of the disease must be completely inhibited. The onset of the disease means that the body has symptoms of the disease.

[0023] The term "prevention" means that before the onset of a disease, a subject is contacted (e.g., administered) with a drug, composition, etc. based on the present application, thereby reducing the symptoms after the onset of the disease compared to not being contacted, and does not mean that the onset of the disease must be completely inhibited.

[0024] In the process of realizing the concept of the present application, it is found that Glp-1 agonists as new drugs for clinical treatment of type 2 diabetes mellitus, the main drugs are liraglutide and semaglutide, their main functions are to activate Glp-1 receptor, promote insulin secretion, inhibit glucagon secretion, delay gastric emptying, and increase satiety. Therefore, they not only have obvious therapeutic effect on the function of regulating blood sugar, but also have significant inhibitory effect on the appetite of obese patients.

[0025] Nitrofurazone is an oral nitrofurazone antibiotic, which belongs to nitrofurazone compounds, and is an effective inhibitor of STAT3, which inhibits the enzyme system of bacteria (such as pyruvate dehydrogenase), interferes with bacterial DNA synthesis, thereby playing a bacteriostatic or bactericidal role. And has anti-cancer and anti-metastatic activity, previous studies have shown that nitrofurazone has important functions in the treatment of intestinal infections, immunomodulation, antiviral, antitumor, etc., but its potential in the treatment of obesity has not been explored.

[0026] Specifically, according to an embodiment of the aspect of the present application, there is provided a use of nitrofurazone or a pharmaceutically acceptable derivative thereof, the use comprising the use of nitrofurazone in the preparation of a medicament for reducing food intake and / or suppressing appetite.

[0027] According to an embodiment of the present application, nitrofurazone can reduce food intake and / or suppress appetite without dependence on blood glucose regulation, without dependence on the gastrointestinal tract, and without causing side effects such as nausea and vomiting; its target of action can not be located in the peripheral nervous system, but can act on the central nervous system; nitrofurazone can significantly reduce the intake of ordinary food and high-fat food by mice, and suppress the appetite of mice, thereby significantly reducing the body weight of obese mice, in a manner of activating Glp1-r. Therefore, nitrofurazone has the potential to treat obesity, and can be used in the preparation of a medicament for treating obesity.

[0028] According to an embodiment of the present application, nitrofurazone reduces food intake and / or suppresses appetite as a Glp1-r receptor agonist.

[0029] According to an embodiment of the present application, nitrofurazone and Glp-1 have similar spatial structures, can bind to Glp-1 receptors to activate Glp1-r, and can be used to treat obesity and suppress the intake of ordinary food and high-fat food by mice without affecting the change in blood glucose, in a manner of reducing the body weight of obese mice. Moreover, the effect of appetite suppression does not change after vagotomy, proving that the effect is not dependent on peripheral gastrointestinal signals.

[0030] According to an embodiment of the present application, the use further comprises the use of nitrofurazone in the preparation of a medicament for treating obesity and / or reducing body weight.

[0031] According to an embodiment of the present application, by reducing food intake and suppressing appetite, nitrofurazone can effectively reduce energy intake, thereby helping to control body weight and treat obesity.

[0032] According to an embodiment of the present application, the use comprises salifying nitrofurazone and / or chemically modifying nitrofurazone.

[0033] According to an embodiment of the present application, by salifying or chemically modifying nitrofurazone, the stability, bioavailability, or efficacy of nitrofurazone can be improved. Salification and chemical modification can improve the pharmacokinetic properties of nitrofurazone, such as prolonging the half-life, enhancing the targeting, or reducing the degradation, thereby improving the effectiveness and convenience of nitrofurazone in practical applications.

[0034] According to an embodiment of the present application, the salt of nitrofurazone comprises at least one of a metal salt of nitrofurazone, a salt of nitrofurazone with an inorganic acid, and a salt of nitrofurazone with an organic acid; and the chemical modification comprises at least one of acetylation, amination, methylation, phosphorylation, glycosylation, lipidation, ubiquitination, biotin labeling, and fluorescent protein labeling.

[0035] According to an embodiment of the present application, the metal salt of nitrofurazone includes at least one of lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, manganese salt, copper salt, zinc salt, aluminum salt; the inorganic acid includes at least one of hydrochloric acid, sulfuric acid, boric acid or carbonic acid; the organic acid includes at least one of acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, oxalic acid.

[0036] According to an embodiment of the present application, the pharmaceutical composition is used for reducing food intake and / or treating obesity; the pharmaceutical composition includes nitrofurazone or a pharmaceutically acceptable derivative thereof as an active ingredient.

[0037] According to an embodiment of the present application, the derivative includes a salt of nitrofurazone and / or a chemical modification of nitrofurazone.

[0038] According to an embodiment of the present application, when the pharmaceutical active ingredient is a pharmaceutically acceptable salt, such as a pharmaceutically acceptable acid addition salt or base addition salt, it can be prepared in situ by reaction with a pH adjusting agent in solution, thereby producing the corresponding free active ingredient, such as a free basic active ingredient or a free acidic active ingredient.

[0039] According to an embodiment of the present application, the dosage form of the pharmaceutical composition is any one of aerosol, spray, powder, pill, tablet, film, ointment, suppository, paste, solution, injection, mixture, lotion or liniment.

[0040] According to an embodiment of the present application, the dosage form of the plurality of drugs meets different treatment needs, improves patient compliance, optimizes the efficacy and safety of the drug, can meet different clinical needs and patient preferences, and adapts to different administration routes.

[0041] Specifically, the pharmaceutical composition can further include an auxiliary ingredient, including at least one of a preservative, a solubilizer, a stabilizer, a diluent, a lubricant, which can improve the physical and chemical properties of the drug, improve the efficacy and safety of the drug, also increase the acceptability of the preparation, meet different clinical needs and patient preferences, while helping the stable release and effective delivery of the drug, and also improve the safety of the preparation. By reasonably selecting and using these auxiliary materials, the dosage form of the drug can be optimized to improve the therapeutic effect.

[0042] According to an embodiment of the present application, the dosage of the pharmaceutical composition is 0.1-100 mg / kg.

[0043] According to the embodiments of the present application, the dosage of the drug can be 0.1 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, but is not limited to the listed values, and other values not listed in the range are also applicable.

[0044] According to the embodiments of the present application, the administration method is preferably intraperitoneal injection, but is not limited to only intraperitoneal injection, and other methods such as oral administration can be adjusted to appropriate dosage forms.

[0045] The embodiments of the present application will be further explained below, and in the following embodiments, if not specifically stated, all are conventional commercially available reagents.

[0046] In the results graphs of the following embodiments, * indicates P value <0.05, ** indicates P value <0.01, and *** indicates P value <0.001, and P value <0.05 is considered to have statistically significant differences in data, and the statistical results are represented by mean value (n is the total number of experimental groups) ± standard error.

[0047] Example 1 Effect of nitrofurazone injection on diet, blood glucose and body weight of mice

[0048] Experimental animals: 8-week-old normal male C57BL / 6J mice were selected as experimental objects and divided into experimental and control groups. 8-week-old normal male mice expressing glucagon-like peptide-1 receptor (Glp1-r) cells were specifically labeled and functionally regulated transgenic mice (i.e. Glp1r-cre::Ai14 transgenic mice) were divided into experimental and control groups.

[0049] Drug preparation: Nitrofurazone was added to 10% dimethyl sulfoxide (DMSO) + 40% polyethylene glycol (PEG)-300 + 5% Tween-80 + 45% saline solvent to prepare a suspension, and the suspension was prepared at an appropriate concentration and used immediately.

[0050] Solvent control group: 10% dimethyl sulfoxide (DMSO) + 40% polyethylene glycol (PEG)-300 + 5% Tween-80 + 45% saline solvent, hereinafter referred to as solvent.

[0051] Intraperitoneal injection experiment: The mice were fixed, and an appropriate amount of nitrofurazone suspension was drawn using a 1 mL syringe, and slowly injected into the abdominal cavity at the lower 1 / 3 of the mouse abdomen, avoiding the internal organs. The dose of nitrofurazone injected intraperitoneally was 50 mg / kg, and the control group was injected with an equal amount of solvent, and the operation steps were the same as the experimental group.

[0052] Food intake measurement: Mice were kept in isolation for 3 days to acclimatize before the formal experiment. During free feeding or after fasting overnight for 12 hours, nifurazolidone or a solvent was injected into the mice, and food intake was monitored at 0h, 0.5h, 1h, 1.5h, 2h, and 5h.

[0053] Blood glucose measurement: Before the formal experiment, the mice were kept alone for 3 days to adapt. They were injected with physiological saline in the peritoneum every day and stroked for 5 minutes. After fasting overnight for 12 hours, they were injected with nifurazolidone or solvent. Blood glucose values ​​were monitored at 0h, 0.5h, 1h, 1.5h and 2h.

[0054] Body weight measurement: Nifurazolidone or its solvent was injected at the same time every day for one week, and the body weight of the mice was recorded at the same time before each injection.

[0055] Experimental Methods: Twelve mice were randomly divided into n=6 pairs. Under normal conditions and after a 12-hour overnight fast, nifurazolidone was administered intraperitoneally, and the mice's food intake was monitored for 5 hours. After a 12-hour overnight fast, nifurazolidone was administered intraperitoneally, and blood glucose levels were monitored for 2 hours. Overnight high-fat food intake was also monitored. Twelve mice were fed a high-fat diet for two weeks to increase their body weight, and then randomly divided into n=6 pairs. Nifurazolidone was administered intraperitoneally at the same time each day for one week, and the daily body weight changes of the obese mice were monitored. Results are shown below. Figures 1-5 As shown.

[0056] Figure 1 This is a graph showing the changes in the intake of ordinary food in mice after intraperitoneal injection of nifurazolidone in Example 1 of the present invention; Figure 2 This is a graph showing the changes in normal food intake in mice after intraperitoneal injection of nifurazolidone following a 12-hour fast in Example 1 of the present invention. Figure 3 This is a graph showing the relative blood glucose changes in mice after intraperitoneal injection of nifurazolidone following a 12-hour fast in Example 1 of the present invention. Figure 4 This is a comparison chart of overnight high-fat food intake in mice after intraperitoneal injection of nifurazolidone in Example 1 of the present invention; Figure 5 This is a graph showing the weight change of obese mice one week after intraperitoneal injection of nifurazolidone in Example 1 of the present invention.

[0057] according to Figures 1-5 It can be seen that after intraperitoneal injection of nifurazolidone, the amount of food consumed by mice was significantly reduced. Figure 1 After fasting mice overnight for 12 hours, mice injected intraperitoneally with nifurazide showed a significant reduction in food intake. Figure 2 ), while blood glucose showed no significant change ( Figure 3 Intraperitoneal injection of nifurazolidone significantly reduced the amount of high-fat food consumed overnight in mice. Figure 4), mice were fed high-fat diet for two weeks, and after obesity, intraperitoneal injection of furazolidone at the same time point for one week, the body weight of obese mice decreased significantly (P < 0.05) Figure 5 ).

[0058] Example 2 Mechanism of furazolidone on diet, blood glucose and body weight of mice

[0059] Experimental animals: 8-week-old normal male C57BL / 6J mice were selected as experimental objects and divided into experimental and control groups. 8-week-old normal male Glp1r-cre::Ai14 transgenic mice were divided into experimental and control groups.

[0060] Drug preparation: Furazolidone was added to 10% DMSO+40% PEG-300+5% Tween-80+45% Saline solvent to prepare a suspension, and the suspension with appropriate concentration was prepared for immediate use.

[0061] Intraperitoneal injection experiment: The mice were fixed, and an appropriate amount of furazolidone suspension was drawn with a 1 mL syringe, and slowly injected into the abdominal cavity at the lower 1 / 3 of the mouse abdomen, avoiding the internal organs. The dose of furazolidone injected intraperitoneally was 50 mg / kg, and the control group was injected with an equal amount of 10% DMSO+40% PEG-300+5% Tween-80+45% Saline solvent, and the operation steps were the same as the experimental group.

[0062] Measurement of food intake: Before the formal experiment, the mice were individually housed for 3 days. During the free feeding of the mice, or after feeding after overnight fasting for 12 hours, furazolidone or solvent was injected, and the food intake at 0h, 0.5h, 1h, 1.5h, 2h, 5h was monitored.

[0063] Measurement of blood glucose: Before the formal experiment, the mice were individually housed for 3 days, and intraperitoneal injection of normal saline was performed every day, and the mice were stroked for 5 minutes, and furazolidone or solvent was injected after overnight fasting for 12 hours, and the blood glucose value at 0h, 0.5h, 1h, 1.5h, 2h was monitored.

[0064] Measurement of body weight: Furazolidone or its solvent was injected at the same time point for one week, and the body weight of the mice was recorded at the same time point before injection of the drug every day.

[0065] Gastroenteric vagotomy: 7-8 hours before surgery, mice need to be fasted. When the operation, through intraperitoneal injection of tribromoethanol solution (ratio: tribromoethanol 0.625g, tertiary amyl alcohol 1.25mL, ultrapure water 50mL; 0.4mL / 20g) deeply anesthetize the mouse, then remove the hair on the abdomen. Wipe the mouse abdomen with alcohol cotton ball, cut a wound of about 2cm along the midline of the abdomen from below the xiphoid cartilage, use forceps to expose the stomach outside the body from the wound. Use a glass microelectrode bend to hook the esophagus, and the subphrenic vagus nerve is attached to the two sides of the esophagus. Use a glass microelectrode bend to hook the bilateral vagus nerve, and use a microscissors to cut off a small piece of vagus nerve about 5mm from the left and right branches of the stomach end to prevent the subphrenic vagus nerve from reconnecting in a short time. The first three days after the operation need to pay close attention to the state of the mouse, and the surviving mice recover for 1 week before the behavior experiment.

[0066] Detection of vomiting behavior: place the mouse in a 2000ml transparent large beaker, record the mouse behavior with a high-resolution camera for 3 hours. In order to reduce the influence of human factors, avoid real-time observation by the experimental operator during the experiment recording, and observe the video material after the recording is completed, and all behavior monitoring is carried out in the same circadian rhythm time. Before the formal experiment, the mice are individually housed for 3 days, intraperitoneally injected with normal saline every day, and stroked for 5 minutes. On the day of behavior monitoring, the mice were transferred from the feeding room to the behavior room 1 hour in advance to adapt to the environment in the room, and the 2000mL large beaker used for monitoring was cleaned with 75% ethanol to eliminate the odor left by other mice.

[0067] Experimental method: 12 mice were randomly divided into 6 groups, 6 mice were subjected to gastroenteric vagotomy, and 6 mice were subjected to sham operation. After recovery, the mice were intraperitoneally injected with furazolidone and the ordinary food intake of the mice was monitored. 16 mice were randomly divided into 8 groups, 8 mice were injected with solvent control and 8 mice were injected with furazolidone, 3 hours of video was recorded, and the number of vomiting and defecation of the mice within 3 hours was observed, and the experimental results are shown in Figures 6-10

[0068] Figure 6 The figure for the ordinary food intake of the mice after intraperitoneal injection of furazolidone after gastroenteric vagotomy in Example 2 of the present application; Figure 7 The figure for the ordinary food intake of the mice after intraperitoneal injection of furazolidone after gastroenteric vagotomy after fasting for 12 hours in Example 2 of the present application; Figure 8 The figure for the number of vomiting within 3 hours after intraperitoneal injection of furazolidone in Example 2 of the present application; Figure 9 The figure for the vomiting latency within 3 hours after intraperitoneal injection of furazolidone in Example 2 of the present application; Figure 10 The figure for the number of defecation within 3 hours after intraperitoneal injection of furazolidone in Example 2 of the present application.​

[0069] According to Figures 6-10 It can be seen that the solvent control group mice and the mice injected with nitrofurazone have no significant difference in the number of vomiting, vomiting latency, and stool frequency, which can rule out that the inhibition of appetite and weight loss of mice by nitrofurazone is caused by drug side effects. Moreover, the inhibition of appetite is not dependent on peripheral gastrointestinal signals, and even if the vagus nerve of the intestine and stomach is cut off, nitrofurazone can still effectively inhibit food intake, proving that it directly regulates appetite through the central nervous system rather than relying on peripheral gastrointestinal pathways. Moreover, it can be seen that nitrofurazone does not cause vomiting or abnormal defecation, which is significantly different from the gastrointestinal adverse reactions of traditional Glp-1 agonists.

[0070] Example 3 Effect of nitrofurazone on mouse brain nerves

[0071] Glp1r-cre::Ai14 transgenic mice (all Glp1-r neurons express red fluorescent protein) were injected intraperitoneally with solvent and nitrofurazone, respectively, to observe the number of marker proteins (c-Fos protein) of neuron activation in the hypothalamic paraventricular nucleus (PVN brain region) regulating food intake. It is speculated that nitrofurazone has a similar spatial structure to Glp-1 and can bind to the Glp-1 receptor, so nitrofurazone is divided into four concentration gradients, and the concentration is detected by an Elisa kit for detecting Glp-1.

[0072] c-Fos protein staining: After high-fat modeling, obese Glp1r-cre::Ai14 transgenic mice were randomly divided into 5 groups, and before the formal experiment, physiological saline was injected intraperitoneally every day, and the mice were stroked for 5 minutes. One hour after intraperitoneal injection of solvent and nitrofurazone, perfusion was performed to take the brain. Then, the brain slices were frozen, the brain slices containing the paraventricular nucleus were selected according to the brain positioning map, 100% Triton X-100 was diluted with 1× phosphate buffered saline (1× PBS) to obtain 0.4% Triton X-100, i.e. phosphate buffered saline containing Triton (PBST), 5% donkey serum was added, and the blocking was performed on a shaker for 1 hour, and then the brain slices were incubated in the first antibody at 4°C for 24 hours. The first antibody is rabbit-derived c-Fos (1:1000), and the brain slices after incubation are placed in the second antibody in a 4°C refrigerator in the dark. The second antibody is a donkey anti-rabbit antibody conjugated with fluorescent dye (Alexa Fluor) 647 (1:500), and then the slices are pasted. The microscopic images of the samples are collected by a Zeiss LSM980 laser confocal system, and the image collection parameters of the experimental group and the control group are set to be completely consistent.

[0073] Enzyme-linked immunosorbent assay (Elisa): nitrofurazone was divided into four concentrations. The solvent was the negative control, and the exenatide was the positive control. The orbital blood was collected to collect three mouse sera, and all were collected in ethylene glycol bis (2-aminoethylether) tetraacetic acid (EGTA) coated tubes, and all samples were then measured and quantified with a glucagon-like peptide-1 enzyme-linked immunosorbent assay kit (Cat. RK15288, ABclonal). All standards and samples were determined in duplicate. The concentration was calculated according to the manufacturer's instructions, and the results are shown in Table 1 below.

[0074] Table 1 Elisa detection results

[0075]

[0076] Figure 11 Figure of the detection results of the hypothalamic paraventricular nucleus (PVN) brain region neuron activation marker protein (c-Fos protein) of the obese Glp1r-cre::Ai14 mice after intraperitoneal injection of nitrofurazone in Example 3 of the present application; Figure 12 Figure of the PVN brain region c-Fos protein of the Glp1r-cre::Ai14 mice in Example 3 of the present application after intraperitoneal injection of nitrofurazone, wherein A is a paraventricular nucleus statistical diagram, and B is a Glp-1 receptor statistical diagram.

[0077] According to Table 1, Figure 11 and Figure 12 It can be seen that the Elisa kit for detecting Glp-1 can detect the concentration of nitrofurazone, although it is lower than the positive control exenatide, but the detected concentration increases with the increase of the concentration of nitrofurazone, and each concentration is higher than the solvent control, which means that nitrofurazone may be similar to the spatial structure of Glp-1, so it can be detected. Figure 11 and Figure 12 It can be seen that nitrofurazone can activate the neuron Glp-1 receptor in the related brain region, so nitrofurazone may be similar to the spatial structure of Glp-1, so as to activate the hypothalamic PVN region c-Fos by activating the neuron Glp-1 receptor to regulate appetite, and confirm that it inhibits appetite through the Glp-1 receptor.

[0078] The above specific embodiments further illustrate the purpose, technical solution and beneficial effects of the present application. It should be understood that the above description is only a specific embodiment of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. Use of nitrofurazone or a pharmaceutically acceptable salt thereof, wherein the use is a use of nitrofurazone in the manufacture of a medicament for reducing food intake and / or suppressing appetite. The nitrofurazone reduces food intake and / or suppresses appetite as a glucagon-like peptide-1-r receptor agonist.

2. Use according to claim 1, characterized in that, 3. Use of nitrofurazone or a pharmaceutically acceptable salt thereof, wherein the use is a use of the nitrofurazone in the manufacture of a medicament for treating obesity and / or reducing body weight.

4. The use according to claim 1 or 3, wherein the pharmaceutically acceptable salt of nitrofurazone includes at least one of a metal salt of nitrofurazone, a salt of nitrofurazone with an inorganic acid, and a salt of nitrofurazone with an organic acid.

5. The use according to claim 4, wherein the metal salt includes at least one of a lithium salt, a sodium salt, a potassium salt, a calcium salt, a magnesium salt, a manganese salt, a copper salt, a zinc salt, and an aluminum salt; the inorganic acid includes hydrochloric acid, sulfuric acid, boric acid, or carbonic acid; and the organic acid includes at least one of acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, and oxalic acid. ​ ​ ​ ​ ​ ​

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