Application of nifurazil or pharmaceutically acceptable derivative thereof and pharmaceutical composition

Abstract

The invention provides application of nifurazide or a pharmaceutically acceptable derivative thereof and a pharmaceutical composition, and belongs to the technical field of biological medicines. The application comprises application of nifurazil in preparation of drugs for reducing food intake and / or inhibiting appetite. The nifurazil does not depend on blood sugar regulation and intestines and stomach, and does not cause side effects of nausea and vomiting; the action target of the nifurazil is not located in the peripheral nervous system but acts on the central nervous system, and the nifurazil can be used for remarkably reducing the intake of common food and high-fat food of mice and inhibiting the appetite of the mice in a mode of activating Glp1-r, so that the weight of the obese mice is remarkably reduced. Therefore, the nifurazil has the potential of treating the obesity and can be applied to preparation of the medicine for treating the obesity.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the application and pharmaceutical composition of nifurozide or its pharmaceutically acceptable derivatives. Background Art

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

[0003] Currently, the treatment of obesity relies primarily on lifestyle interventions, medications, and metabolic surgery. Lifestyle interventions (such as diet, exercise, and behavioral therapy) are foundational approaches, but long-term compliance is poor and their effectiveness in patients with moderate to severe obesity is limited. Regarding medications, existing drugs such as glucagon-like peptide-1 (Glp-1) receptor agonists (such as semaglutide) and pancreatic lipase inhibitors (such as orlistat) can reduce weight, but they are associated with side effects (such as hypoglycemia, gastrointestinal reactions, and the risk of pancreatitis) and weight rebound after discontinuation. Metabolic surgery (such as gastric bypass) can significantly reduce weight and improve metabolic abnormalities, but it carries high surgical risks, is expensive, and may lead to malnutrition or postoperative complications, making it suitable only for a minority of patients. These limitations highlight the urgent need for the development of new anti-obesity drugs. Ideally, these drugs should offer improved safety, more significant weight loss, and the multiple benefits of improving metabolic markers. Summary of the Invention

[0004] In view of this, in order to at least partially solve at least one of the above-mentioned technical problems, the present invention provides a use of nifurozide or a pharmaceutically acceptable derivative thereof, including the use of nifurozide in the preparation of a drug for reducing food intake and / or suppressing appetite.

[0005] According to one embodiment of the present invention, a pharmaceutical composition is provided, which is used for reducing food intake and / or treating obesity; the pharmaceutical composition comprises nifurozide or a pharmaceutically acceptable derivative thereof as an active ingredient.

[0006] According to embodiments of the present invention, nifurozide is independent of blood sugar regulation and gastrointestinal function, and does not cause nausea and vomiting. Its target, rather than the peripheral nervous system, acts on the central nervous system. Nifurozide may significantly reduce mice's intake of both regular and high-fat foods by activating Glp1-r, suppressing their appetite and thus significantly reducing the weight of obese mice. Therefore, nifurozide has the potential to treat obesity and can be used in the preparation of drugs for the treatment of obesity. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1This is a graph showing changes in normal food intake in mice after intraperitoneal injection of nifurozide in Example 1 of the present invention;

[0008] Figure 2 This is a graph showing changes in normal food intake in mice after intraperitoneal injection of nifurozide after fasting for 12 hours according to Example 1 of the present invention;

[0009] Figure 3 This is a graph showing the relative blood sugar changes in mice after intraperitoneal injection of nifurozide after fasting for 12 hours in Example 1 of the present invention;

[0010] Figure 4 This is a comparison chart of overnight high-fat food intake in mice after intraperitoneal injection of nifurozide in Example 1 of the present invention;

[0011] Figure 5 This is a graph showing the weight changes of obese mice one week after intraperitoneal injection of nifurozide in Example 1 of the present invention;

[0012] Figure 6 This is a statistical chart of normal food intake in mice after intraperitoneal injection of nifurozide after gastrointestinal vagus nerve removal in Example 2 of the present invention;

[0013] Figure 7 This is a statistical chart of normal food intake in mice after fasting for 12 hours and intraperitoneal injection of nifurozide after removal of the gastrointestinal vagus nerve in Example 2 of the present invention;

[0014] Figure 8 This is a statistical chart showing the number of dry heaves within 3 hours after intraperitoneal injection of nifurozide in mice according to Example 2 of the present invention;

[0015] Figure 9 This is a statistical chart of the latency of retching within 3 hours after intraperitoneal injection of nifurozide in mice according to Example 2 of the present invention;

[0016] Figure 10 This is a statistical chart of the number of bowel movements within 3 hours after intraperitoneal injection of nifurozide in mice according to Example 2 of the present invention;

[0017] Figure 11 This is a graph showing the results of detecting a marker protein (c-Fos protein) for neuronal activation in the paraventricular nucleus (PVN) of the hypothalamus of obese Glp1r-cre::Ai14 mice induced by high-fat diet after intraperitoneal injection of nifurozide in Example 3 of the present invention;

[0018] Figure 12 This is a statistical diagram of c-Fos protein in the PVN brain region of Glp1r-cre::Ai14 mice injected intraperitoneally with nifurozide in Example 3 of the present invention, wherein A is a statistical diagram of the paraventricular nucleus and B is a statistical diagram of the Glp-1 receptor. DETAILED DESCRIPTION

[0019] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. However, it should be understood that these descriptions are exemplary only and are not intended to limit the scope of the present invention. In the following detailed description, for ease of explanation, many specific details are set forth to provide a comprehensive understanding of embodiments of the present invention. However, it is apparent that one or more embodiments may also be implemented without these specific details. In addition, in the following description, descriptions of known structures and technologies are omitted to avoid unnecessarily confusing the concept of the present invention.

[0020] The terms used herein are only for describing specific embodiments and are not intended to limit the present invention. The term "comprising" used herein indicates the existence of features, steps, operations, but does not exclude the existence or addition of one or more other features.

[0021] In the present invention, the term "pharmaceutically acceptable" refers to compounds, substances, compositions and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, and are commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment.

[0022] The term "treatment" refers to exposing a subject to (e.g., administering) a drug, composition, or the like according to the present invention after the subject has contracted a disease, thereby alleviating the symptoms of the disease compared to a patient without such exposure. It does not necessarily mean that the symptoms of the disease are completely suppressed. Contracting a disease means that the body has developed symptoms of the disease.

[0023] The term "prevention" means that the subject is exposed to (e.g., administered) the drug, composition, etc. according to the present invention before the disease occurs, thereby reducing the symptoms after the disease occurs compared to when the subject does not have such exposure. It does not necessarily mean that the disease must be completely suppressed.

[0024] During the implementation of the present invention, it was discovered that Glp-1 agonists, primarily liraglutide and semaglutide, are novel clinical treatments for type 2 diabetes. Their primary function is to activate the Glp-1 receptor, promoting insulin secretion, inhibiting glucagon secretion, delaying gastric emptying, and increasing satiety. Therefore, they not only have significant therapeutic effects in regulating blood sugar, but also significantly suppress appetite in obese patients.

[0025] Nifuroxide is an oral nitrofuran antibiotic. It is a potent inhibitor of STAT3, inhibiting bacterial enzymes (such as pyruvate dehydrogenase) and interfering with bacterial DNA synthesis, thereby exerting antibacterial or bactericidal effects. It also exhibits anticancer and anti-metastatic activities. Previous studies have shown that nifuroxide has important functions in the treatment of intestinal infections, immunomodulation, antiviral, and anti-tumor effects, but its potential in the treatment of obesity has not been explored.

[0026] Specifically, according to one embodiment of the present invention, there is provided a use of nifurozide or a pharmaceutically acceptable derivative thereof, including use of nifurozide in the preparation of a drug for reducing food intake and / or suppressing appetite.

[0027] According to embodiments of the present invention, nifurozide is independent of blood sugar regulation and gastrointestinal function, and does not cause nausea and vomiting. Its target may not be located in the peripheral nervous system, but rather in the central nervous system. Nifurozide may significantly reduce mice's intake of both regular and high-fat foods by activating Glp1-r, suppressing their appetite and thus significantly reducing the weight of obese mice. Therefore, nifurozide has the potential to treat obesity and can be used in the preparation of drugs for the treatment of obesity.

[0028] According to an embodiment of the present invention, nifurozide acts as a Glp1-r receptor agonist to reduce food intake and / or suppress appetite.

[0029] According to embodiments of the present invention, nifurozide, which has a similar spatial structure to Glp-1, can bind to the Glp-1 receptor to activate Glp1-r, leading to weight loss in obese mice. Nifurozide can be used to treat obesity, inhibiting both regular and high-fat food intake in mice without affecting blood sugar levels. Furthermore, its appetite-suppressing effects persist after vagotomy, demonstrating that its effects are independent of peripheral gastrointestinal signals.

[0030] According to an embodiment of the present invention, the application also includes the use of nifurozide as a drug for treating obesity and / or reducing weight.

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

[0032] According to an embodiment of the present invention, the application includes salification of nifurozide and / or chemical modification of nifurozide.

[0033] According to embodiments of the present invention, nifurozide is salified or chemically modified to improve its stability, bioavailability, or efficacy. Salt formation and chemical modification can improve the pharmacokinetic properties of nifurozide, such as extending its half-life, enhancing its targeting, or reducing its degradation, thereby enhancing its effectiveness and convenience in practical applications.

[0034] According to an embodiment of the present invention, the salt of nifurozide includes at least one of a metal salt of nifurozide, a salt of nifurozide and an inorganic acid, and a salt of nifurozide and an organic acid; and the chemical modification includes 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 invention, the metal salt of nifuroxide includes at least one of lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, manganese salt, copper salt, zinc salt, and 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.

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

[0037] According to an embodiment of the present invention, the derivative includes a salt of nifuroxide and / or a chemically modified substance of nifuroxide.

[0038] According to an embodiment of the present invention, 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 reacted with a pH regulator in the solution to thereby prepare in situ the corresponding free active ingredient, such as a free alkaline active ingredient or a free acidic active ingredient.

[0039] According to an embodiment of the present invention, 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 the embodiments of the present invention, the dosage forms of multiple drugs meet different treatment needs, improve patient compliance, optimize the efficacy and safety of drugs, can meet different clinical needs and patient preferences, and adapt to different administration routes.

[0041] Specifically, pharmaceutical compositions may also include excipients, including at least one of a preservative, solubilizer, stabilizer, diluent, and lubricant. These excipients can improve the drug's physical and chemical properties, enhance its efficacy and safety, increase its acceptability, meet diverse clinical needs and patient preferences, and contribute to its stable release and effective delivery, while also improving its safety. By rationally selecting and using these excipients, the drug's dosage form can be optimized, enhancing its therapeutic efficacy.

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

[0043] According to an embodiment of the present invention, the dosage 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 unlisted values ​​within the numerical range are also applicable.

[0044] According to an embodiment of the present invention, the administration method is preferably intraperitoneal injection, but is not limited to intraperitoneal injection. The drug may be administered orally or in other ways by adjusting the appropriate dosage form.

[0045] The present invention will be further explained below with reference to specific embodiments. In the following embodiments, conventional commercial reagents are used unless otherwise stated.

[0046] In the following implementation plan result graph, * indicates P value <0.05, ** indicates P value <0.01, and *** indicates P value <0.001. The data are considered to have statistically significant differences only when P value <0.05. The data values ​​are expressed as mean (n is the total number of experimental groups) ± standard error in the statistical result graph.

[0047] Example 1 Effects of Nifurozide Injection on Diet, Blood Glucose and Body Weight in Mice

[0048] Experimental Animals: 8-week-old, normally developed male C57BL / 6J mice were used as experimental subjects and divided into experimental and control groups. 8-week-old, normally developed male transgenic mice that specifically label and functionally regulate cells expressing the glucagon-like peptide-1 receptor (Glp1-r) (i.e., Glp1r-cre::Ai14 transgenic mice) were also divided into experimental and control groups.

[0049] Drug preparation: Add nifurozide to 10% dimethyl sulfoxide (DMSO) + 40% polyethylene glycol (PEG)-300 + 5% Tween-80 + 45% saline solvent to prepare a suspension. Prepare a suspension of appropriate concentration and prepare it before use.

[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: Immobilize the mouse and use a 1 mL syringe to draw an appropriate amount of nifurozide suspension. Slowly inject into the peritoneal cavity at the lower third of the mouse's abdomen, avoiding internal organs. The intraperitoneal dose of nifurozide is 50 mg / kg. The control group receives an equal amount of solvent. The procedure is the same as for the experimental group.

[0052] Measurement of food intake: Before the formal experiment, mice were individually housed and acclimated for 3 days. Nifurozide or the solvent was injected into the mice while they were eating freely or after a 12-hour overnight fast and then fed. Food intake was monitored at 0, 0.5, 1, 1.5, 2, and 5 hours.

[0053] Blood glucose measurement: Before the formal experiment, mice were individually housed for 3 days for adaptation. Normal saline was injected intraperitoneally every day and the mice were stroked for 5 minutes. After fasting for 12 hours overnight, nifurozide or solvent was injected, and blood glucose levels were monitored at 0h, 0.5h, 1h, 1.5h, and 2h.

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

[0055] Experimental method: 12 mice were randomly divided into 6 pairs of 6. After normal conditions and overnight fasting for 12 hours, the mice were intraperitoneally injected with nifuroxide and their food intake within 5 hours was monitored. After overnight fasting for 12 hours, the mice were intraperitoneally injected with nifuroxide and their blood sugar changes within 2 hours were monitored. Overnight high-fat food intake of mice. After 12 mice were fed a high-fat diet for two weeks to increase their weight, they were randomly divided into 6 pairs of 6. Nifuroxide was intraperitoneally injected at the same time every day for one week. The weight changes of obese mice were monitored every day for one week. The results are as follows. Figures 1 to 5 shown.

[0056] Figure 1 This is a graph showing changes in normal food intake in mice after intraperitoneal injection of nifurozide in Example 1 of the present invention; Figure 2 This is a graph showing changes in normal food intake in mice after intraperitoneal injection of nifurozide after fasting for 12 hours according to Example 1 of the present invention; Figure 3 This is a graph showing the relative blood sugar changes in mice after intraperitoneal injection of nifurozide after fasting for 12 hours 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 nifurozide in Example 1 of the present invention; Figure 5 This is a graph showing the weight changes of obese mice one week after intraperitoneal injection of nifurozide according to Example 1 of the present invention.

[0057] according to Figures 1 to 5 It can be seen that after intraperitoneal injection of nifuroxide, the food intake of mice was significantly reduced ( Figure 1 ), after the mice were fasted for 12 hours overnight, the food intake of mice injected intraperitoneally with nifuroxide was significantly reduced ( Figure 2 ), while blood sugar showed no significant changes ( Figure 3 After intraperitoneal injection of nifurozide, the mice's overnight food intake on high-fat food was significantly reduced ( Figure 4), mice were fed a high-fat diet for two weeks, and then nifuroxide was injected intraperitoneally at the same time every day for one week after becoming obese. The weight of the obese mice decreased significantly ( Figure 5 ).

[0058] Example 2 Mechanism of the Effect of Nifurozide on Diet, Blood Glucose and Body Weight in Mice

[0059] Experimental Animals: 8-week-old male C57BL / 6J mice with normal development were used as experimental and control groups. 8-week-old male Glp1r-cre::Ai14 transgenic mice with normal development were also divided into experimental and control groups.

[0060] Drug preparation: Add nifurozide to 10% DMSO + 40% PEG-300 + 5% Tween-80 + 45% Saline solvent to prepare a suspension. Prepare a suspension of appropriate concentration and prepare it before use.

[0061] Intraperitoneal injection experiment: Immobilize the mouse and slowly inject an appropriate amount of nifurozide suspension into the peritoneal cavity using a 1 mL syringe into the lower third of the mouse's abdomen, avoiding internal organs. The intraperitoneal dose of nifurozide is 50 mg / kg. The control group receives an equivalent volume of 10% DMSO, 40% PEG-300, 5% Tween-80, and 45% Saline solution. The procedure is the same as for the experimental group.

[0062] Measurement of food intake: Before the formal experiment, mice were individually housed and acclimated for 3 days. Nifurozide or the solvent was injected into the mice while they were eating freely or after a 12-hour overnight fast and then fed. Food intake was monitored at 0, 0.5, 1, 1.5, 2, and 5 hours.

[0063] Blood glucose measurement: Before the formal experiment, mice were individually housed for 3 days for adaptation. Normal saline was injected intraperitoneally every day and the mice were stroked for 5 minutes. After fasting for 12 hours overnight, nifurozide or solvent was injected, and blood glucose levels were monitored at 0h, 0.5h, 1h, 1.5h, and 2h.

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

[0065] Gastrointestinal vagus nerve removal: The mouse must be fasted for 7–8 hours before surgery. During surgery, the mouse is deeply anesthetized with an intraperitoneal injection of a tribromoethanol solution (ratio: 0.625 g tribromoethanol, 1.25 mL tert-amyl alcohol, 50 mL ultrapure water; 0.4 mL / 20 g). The abdominal hair is then shaved. The mouse's abdomen is cleaned with an alcohol swab. A 2 cm incision is made along the midline of the abdomen, starting just below the xiphoid cartilage. The stomach is then freed from the incision with forceps and exposed to the outside of the body. The esophagus is delineated using a curved glass microelectrode. The subphrenic vagus nerve attaches to both sides of the esophagus. The vagus nerves are delineated bilaterally using a curved glass microelectrode. Microscissors are used to cut a short segment of the vagus nerve, approximately 5 mm from the left and right branches at the stomach end, to prevent reconnection within a short period of time. The mouse's condition should be closely monitored for the first three days after surgery. Surviving mice will undergo behavioral testing after one week of recovery.

[0066] Detection of retching behavior: The mice were placed in a 2000ml transparent beaker, and the behavior of the mice was recorded with a high-resolution camera for 3 hours. In order to reduce the influence of human factors, the experimental operators were prevented from real-time observation during the experimental recording. The video materials were used for observation after the recording, and all behavioral monitoring was carried out within the same circadian rhythm time. Before the formal experiment, the mice were housed individually for 3 days, and normal saline was injected intraperitoneally every day, and the mice were stroked for 5 minutes. On the day of behavioral monitoring, the mice were transferred from the feeding room to the behavioral room 1 hour in advance to adapt to the environment in the room. The 2000mL 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 pairs of 6. 6 mice underwent gastrointestinal vagotomy and 6 mice underwent sham surgery. After recovery, nifurozide was injected intraperitoneally and the mice's normal food intake was monitored. 16 mice were randomly divided into 8 pairs of 8 and injected with solvent control and nifurozide respectively. Videos were recorded for 3 hours and the number of retching and defecation of the mice was observed within 3 hours. The experimental results are as follows: Figures 6 to 10 shown.

[0068] Figure 6 This is a statistical chart of normal food intake in mice after intraperitoneal injection of nifurozide after gastrointestinal vagus nerve removal in Example 2 of the present invention; Figure 7 This is a statistical chart of normal food intake in mice after fasting for 12 hours and intraperitoneal injection of nifurozide after removal of the gastrointestinal vagus nerve in Example 2 of the present invention; Figure 8 This is a statistical chart showing the number of dry heaves within 3 hours after intraperitoneal injection of nifurozide in mice according to Example 2 of the present invention; Figure 9 This is a statistical chart of the latency of retching within 3 hours after intraperitoneal injection of nifurozide in mice according to Example 2 of the present invention; Figure 10 This is a statistical chart showing the number of bowel movements within 3 hours after intraperitoneal injection of nifurozide in mice according to Example 2 of the present invention.

[0069] according to Figures 6 to 10 As can be seen, there was no significant difference in the number of vomiting episodes, vomiting latency, or bowel movement between the solvent control group and the nifurozide-injected mice, ruling out that the appetite suppression and weight loss effects of nifurozide in mice were not due to drug side effects. Furthermore, appetite suppression is independent of peripheral gastrointestinal signals. Even after vagal resection, nifurozide still effectively suppresses food intake, demonstrating that it directly regulates appetite through the central nervous system, rather than relying on peripheral gastrointestinal pathways. Furthermore, it can be seen that nifurozide did not induce retching or abnormal bowel movements, significantly different from the gastrointestinal adverse reactions associated with traditional Glp-1 agonists.

[0070] Example 3 Effects of Nifurozide on Mouse Brain Nerves

[0071] Glp1r-cre::Ai14 transgenic mice (all Glp1-r neurons express red fluorescent protein) were intraperitoneally injected with solvent and nifurozide. The number of c-Fos, a marker for activation of neurons regulating feeding, in the paraventricular nucleus (PVN) of the hypothalamus was measured. Nifurozide was hypothesized to have similar spatial structures to Glp-1 and could bind to the Glp-1 receptor. Therefore, nifurozide was administered in four concentration gradients and measured using an ELISA kit for Glp-1.

[0072] c-Fos protein staining: Obese Glp1r-cre::Ai14 transgenic mice induced by high-fat diet were randomly divided into pairs of five and five. Before the experiment, mice were intraperitoneally injected with saline and handled for 5 minutes daily. One hour after intraperitoneal injection of vehicle and nifurozide, the brains were perfused and removed. Cryosections were then performed. Brain slices containing the paraventricular nucleus were identified based on a brain 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 with Triton (PBST). 5% donkey serum was then added and permeabilized and blocked on a rocking platform for 1 hour. The slices were then incubated in the primary antibody, rabbit c-Fos (1:1000), at 4°C for 24 hours. After incubation, the slices were incubated in the secondary antibody at 4°C in a dark refrigerator. The secondary antibody was donkey anti-rabbit conjugated with Alexa Fluor 647 (1:500), followed by mounting. Microscopic images of the samples were acquired using a Zeiss LSM980 laser confocal system, and the image acquisition parameters for the experimental and control groups were exactly the same.

[0073] Enzyme-linked immunosorbent assay (ELISA): Nifurozide was assayed at four concentrations. The solvent served as a negative control, and belaglutide served as a positive control. Three mouse sera were collected by orbital bleeding and collected in ethylene glycol bis(2-aminoethyl ether) tetraacetic acid (EGTA)-coated tubes. All samples were then measured and quantified using a glucagon-like peptide-1 enzyme-linked immunosorbent assay kit (Cat. RK15288, ABclonal). All standards and samples were assayed in duplicate. Concentrations were calculated according to the manufacturer's instructions, and the results are shown in Table 1.

[0074] Table 1 Elisa test results

[0075]

[0076] Figure 11 This is a graph showing the results of detecting a marker protein (c-Fos protein) for neuronal activation in the paraventricular nucleus (PVN) of the hypothalamus of obese Glp1r-cre::Ai14 mice induced by high-fat diet after intraperitoneal injection of nifurozide in Example 3 of the present invention; Figure 12 This is a statistical diagram of c-Fos protein in the PVN brain region of Glp1r-cre::Ai14 mice injected intraperitoneally with nifurozide in Example 3 of the present invention, wherein A is a statistical diagram of the paraventricular nucleus and B is a statistical diagram of the Glp-1 receptor.

[0077] According to Table 1, Figure 11 and Figure 12 It can be seen that the concentration of nifurozide can be detected by the Elisa kit for detecting Glp-1. Although it is lower than the positive control belaglutide, the concentration detected increases with the increase of nifurozide concentration. All concentrations are higher than the solvent control, which means that nifurozide may have a similar spatial structure to Glp-1 and thus be detected. Figure 11 and Figure 12 It can be seen that nifurozide can activate neuronal Glp-1 receptors in relevant brain areas. Therefore, nifurozide may have a similar spatial structure to Glp-1, thereby regulating appetite by activating neuronal Glp-1 receptors and activating c-Fos in the hypothalamic PVN area, confirming that it suppresses appetite through Glp-1 receptors.

[0078] The specific embodiments described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A use of nifurozide or a pharmaceutically acceptable derivative thereof, characterized in that: The application includes the application of nifurozide in preparing drugs for reducing food intake and / or suppressing appetite.

2. The use according to claim 1, characterized in that Nifurozide acts as a glucagon-like peptide-1-r receptor agonist to reduce food intake and / or suppress appetite.

3. The use according to claim 1, characterized in that The application also includes the application of nifurozide as a drug for treating obesity and / or reducing weight.

4. The use according to claim 1, characterized in that The application includes salifying the nifuroxide and / or chemically modifying the nifuroxide.

5. The use according to claim 4, characterized in that The salt of nifurozide includes at least one of a metal salt of nifurozide, a salt of nifurozide with an inorganic acid, and a salt of nifurozide with an organic acid; The chemical modification includes at least one of acetylation, amination, methylation, phosphorylation, glycosylation, liposylation, ubiquitination, biotin labeling, and fluorescent protein labeling.

6. The use according to claim 5, characterized in that The metal salt includes at least one of lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, manganese salt, copper salt, zinc salt and aluminum salt; The inorganic acid includes 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, and oxalic acid.

7. A pharmaceutical composition, characterized in that The pharmaceutical composition is used for reducing food intake and / or treating obesity; The pharmaceutical composition comprises nifurozide or a pharmaceutically acceptable derivative thereof as an active ingredient.

8. The pharmaceutical composition according to claim 7, characterized in that The derivatives include salts of nifuroxide and / or chemically modified products of nifuroxide.

9. The pharmaceutical composition according to claim 7, characterized in that 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.

10. The pharmaceutical composition according to claim 7, characterized in that The dosage of the pharmaceutical composition is 0.1-100 mg / kg.

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