Application of GLP-1 receptor stimulant in preparation of drug for reducing defensive attack behavior

Abstract

The invention belongs to the technical field of neurobiology and medicine, and relates to an application of a GLP-1 receptor stimulant in preparation of a drug for reducing defensive aggressive behaviors. Experiments prove that the GLP-1 receptor stimulant can significantly reduce defensive attack behaviors. The GLP-1 receptor stimulant provided by the invention is used as a conventional hypoglycemic drug, has a mature commercially available product, and has the characteristics of safety, high efficiency and low side effect when being applied to preparation of drugs for reducing defensive attack behaviors.

Description

Technical Field

[0001] The present invention belongs to the fields of neurobiology and pharmaceutical technology. Specifically, it relates to the application of a GLP-1 receptor agonist in the preparation of a drug for reducing defensive attack behavior. Background Art

[0002] Defensive attack is an innate behavior induced by environmental threats and is of great significance for the survival of individuals. Existing studies have shown that multiple brain regions, such as the periaqueductal gray (PAG), hypothalamus, etc., play key roles in the regulation of defensive attack behavior. However, the current regulatory mechanism for defensive attack behavior is still not fully understood, and there is a lack of effective intervention measures.

[0003] The glucagon-like peptide-1 receptor (GLP-1R) is a known neuroendocrine regulatory factor that mainly participates in the metabolic regulation of the body, such as regulating appetite, insulin secretion, etc., and is mainly used for the treatment of diabetes and obesity. It regulates blood sugar and body weight by mimicking the action of the natural hormone GLP-1 in the human body and activating related receptors. Existing studies mainly focus on the application of GLP-1R in appetite control and mood disorders, while the research in the field of defensive attack behavior is still relatively blank. In addition, the current treatment methods for abnormal aggressive behavior are limited and mostly focus on traditional psychiatric drugs, but these drugs often have large side effects, such as sedation, cognitive impairment, etc.

[0004] Therefore, developing other drug intervention measures and finding safer and more effective prevention and treatment drugs have important clinical value. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide the application of a GLP-1 receptor agonist in the preparation of a drug for reducing defensive attack behavior.

[0006] To solve the above problems, the technical solution adopted by the present invention is: The present invention provides the application of a GLP-1 receptor agonist in the preparation of a drug for reducing defensive attack behavior.

[0007] As a further improvement of the present invention, the GLP-1 receptor agonist includes exenatide, benaglutide, lixisenatide, liraglutide, dulaglutide, semaglutide, pegolotide, albiglutide, and tirzepatide.

[0008] As a further improvement of the present invention, the defensive aggressive behaviors include defensive aggressive behaviors related to mental diseases and / or organic brain injuries; the mental diseases include, but are not limited to, post-traumatic stress disorder, autism spectrum disorder, social anxiety disorder, borderline personality disorder, bipolar disorder, and Alzheimer's disease.

[0009] As a further improvement of the present invention, the defensive aggressive behaviors include defensive aggressive behaviors related to post-traumatic stress disorder, schizophrenia, autism spectrum disorder, social anxiety disorder, borderline personality disorder, bipolar disorder, Alzheimer's disease, and / or organic brain injuries.

[0010] As a further improvement of the present invention, the GLP-1 receptor agonist is administered via an oral, subcutaneous injection, intravenous injection, or nasal administration route.

[0011] As a further improvement of the present invention, the drug is formulated into one of the following dosage forms: tablets, capsules, granules, powders, suspensions, emulsions, powders, solutions, gels, syrups, pills, tinctures, medicated wines, decoction extracts, lozenges, mixtures, suppositories, injections, inhalants, or sprays.

[0012] As a further improvement of the present invention, it further comprises at least one pharmaceutically acceptable carrier or excipient.

[0013] In the preparation of the drug, the GLP-1 receptor agonist can be administered independently or in combination with a pharmaceutically acceptable carrier and excipient. "Pharmaceutically acceptable carrier or excipient" includes: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifying agents, sweetening agents, flavoring agents, taste masking agents, coloring agents, anti-caking agents, humectants, chelating agents, plasticizers, thickening agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. Those skilled in the art will understand that certain pharmaceutically acceptable excipients can be used with more than one function and with alternative functions, depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation. There are many resources available to those skilled in the art that describe pharmaceutically acceptable excipients and can be used to select suitable pharmaceutically acceptable excipients, such as books like "Remington: The Science and Practice of Pharmacy", "Chinese Pharmaceutical Yearbook", "Pharmaceutics", etc.

[0014] The beneficial effects obtained by adopting the above technical solutions are as follows: 1. The present invention provides a new use, that is, the application of GLP-1 receptor agonist (GLP-1RA) in reducing defensive aggressive behaviors, and it is first demonstrated that GLP-1RA can significantly inhibit the defensive aggressive behaviors of mice, providing a new treatment strategy for the intervention of abnormal aggressive behaviors.

[0015] 2. Traditional anti - aggression drugs are mostly psychiatric drugs (such as antipsychotic drugs and antidepressant drugs), which have relatively large side effects. While GLP - 1RA, as an approved hypoglycemic drug, has high safety. At the same time, as a commonly used hypoglycemic drug, GLP - 1RA already has mature commercial products. Therefore, applying it as an anti - aggression drug in clinical practice greatly improves the efficiency of the drug from research and development to clinical use. Description of the Drawings

[0016] Figure 1 is the experimental flow chart of Example 1 of the present invention; Figure 2 is the open - field test result of Example 1 of the present invention. Among them, A is the test result of male mice; B is the test result of female mice; Figure 3 is the test result of the defensive attack behavior of male mice in Example 1 of the present invention. Among them, A is the latency of the first attack; B is the number of attacks; C is the attack time; Figure 4 is the test result of the defensive attack behavior of female mice in Example 1 of the present invention. Among them, A is the latency of the first attack; B is the number of attacks; C is the attack time; Figure 5 is the experimental flow chart of Example 2 of the present invention; Figure 6 is the test result of the defensive attack behavior of male mice in Example 2 of the present invention. Among them, A is the latency of the first attack; B is the number of attacks; C is the attack time; Figure 7 is the test result of the defensive attack behavior of female mice in Example 2 of the present invention. Among them, A is the latency of the first attack; B is the number of attacks; C is the attack time; Figure 8 is the experimental flow chart of Example 3 of the present invention; Figure 9 is the open - field test result of Example 3 of the present invention. Among them, A is the total moving distance; B is the residence time in the central area; Figure 10 is the test result of the defensive attack behavior of Example 3 of the present invention. Among them, A is the latency of the first attack; B is the number of attacks; C is the attack time. Detailed Embodiments

[0017] To make the objectives, technical solutions and advantages of the present invention clearer, the following describes the invention clearly and completely in conjunction with specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts fall within the scope of protection of the present invention.

[0018] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which this invention belongs. The terms used in the description of the present invention in this specification are only for the purpose of describing specific embodiments, and are not intended to limit the present invention. The experimental methods or test methods involved in the embodiments of the present invention, unless otherwise specified, are conventional methods in the prior art, and their names and / or abbreviations are all conventional names in the field, and are very clear and definite in the relevant application fields. Those skilled in the art can understand the conventional process steps according to the name and apply the corresponding equipment, and implement them under conventional conditions or conditions recommended by the manufacturer. There is no special limitation on the sources of various instruments, equipment, raw materials or reagents used in the embodiments of the present invention. They are all conventional products that can be obtained through regular commercial channels, and can also be prepared according to the conventional methods well-known to those skilled in the art.

[0019] All experiments were approved by the Local Committee for the Care, Use and Protection of Animals at Hebei Medical University and were conducted in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.

[0020] Preparation Example 1 1. Select 7-week-old male ICR mice and 7-week-old female ICR mice (Beijing Vital River Laboratory Animal Technology Co., Ltd.). After one week of adaptive cultivation, the mice were anesthetized with a mixture of anesthetic, muscle relaxant, analgesic and sedative. Using stereotactic surgery, a catheter was implanted into the lateral ventricle of each mouse (AP = -0.7, ML = -1, DV = -2.2) with reference to the brain atlas. The catheter was fixed with dental cement and the mice were housed under conditions of a temperature of 20 - 25°C, a humidity of 50% - 55%, and a 12h light / 12h dark cycle. After 7 days of recovery, they had free access to water and food.

[0021] 2. GLP-1 receptor agonist (GLP-1RA): Liraglutide was dissolved in 0.9% sterile saline solution at a final concentration of 0.5 μg / μL and freshly prepared before administration. During administration, an ICV injection (intracerebroventricular injection) was performed using a Hamilton syringe (25 μL), and the injection volume was 2 μL, which was completed within 10 minutes.

[0022] Example 1: Effect of Acute GLP-1RA Exposure on Defensive Aggressive Behavior in Mice Twenty male mice (n = 20) and twenty female mice (n = 20) selected from Preparation Example 1 were randomly divided into a male control group (n = 10), a male experimental group (n = 10), a female control group (n = 10), and a female experimental group (n = 10). 1. Open Field Test (OFT): Twenty minutes before the Open Field Test, each mouse in the male experimental group and the female experimental group was intracerebroventricularly injected with a GLP-1 receptor agonist (Liraglutide, 0.5 μg / μL, ICV) at a dose of 2 μL / mouse; each mouse in the male control group and the female control group was intracerebroventricularly injected with normal saline at a dose of 2 μL / mouse. The plexiglass arena (40 cm × 40 cm × 40 cm) was divided into a central area (20 cm × 20 cm) and a peripheral area. At the start of the experiment, the test mice were placed in the central area and allowed to freely explore for 5 minutes. The variables measured were the time the mice stayed in the central area and the total distance traveled. Automatic recording was performed using a video tracking system and analysis was carried out using software. After the experiment, the mice were returned to the breeding cage and the plexiglass arena was cleaned with 75% alcohol to remove residual odors.

[0023] The experimental results were as Figure 2 shown. There were no significant differences between the results of the experimental group and the Open Field Test results of the control group (p > 0.05), indicating that GLP-1RA does not affect the locomotor ability or anxiety level of mice.

[0024] 2. Defensive Attack Test (DAT) On the second day after the Open Field Test, twenty minutes before the start of the Defensive Attack Test, the mice used in Step 1 were continued to be used. Each mouse in the male experimental group and the female experimental group was intracerebroventricularly injected with a GLP-1 receptor agonist (Liraglutide, 0.5 μg / μL, ICV) at a dose of 2 μL / mouse; each mouse in the male control group and the female control group was intracerebroventricularly injected with normal saline at a dose of 2 μL / mouse. A plastic model snake equipped with a mechanical stimulation device and a mouse were placed in the same breeding cage. The model snake (weighing 150 grams) was equipped with a crocodile clip shaped like a head (weighing 3 grams). The crocodile clip was used to clamp the mouse's tail to apply continuous harmful mechanical stimulation to the mouse. During the experiment, due to the weight of the fake snake, the mouse could not effectively move to avoid the harmful stimulation. The fake snake was clamped on the mouse's tail for 60 seconds to induce the mouse's defensive attack through mechanical tail clamping stimulation (simulating a threat). The latency of the mouse's first bite, the number of bites, and the total bite duration were recorded. The experiment lasted for 1 minute. After the experiment, the model snake was removed, disinfected with alcohol, and the mouse was returned to the original breeding cage; the experiment was repeated to test the defensive attack behavior of all mice, and the latency of their second bite, the number of bites, and the total bite duration were recorded.

[0025] The experimental results are as Figure 3 、 Figure 4 shown. The defensive attack behavior of the experimental group mice was significantly reduced compared with that of the control group mice. The latency of the first bite was significantly prolonged, the number of bites was significantly reduced, and at the same time, the total bite duration was reduced.

[0026] Example 2: The action time of a single injection of GLP-1RA in mice.

[0027] Twenty male mice (n = 20) and twenty female mice (n = 20) selected from Preparation Example 1 were randomly divided into a male control group (n = 10), a male experimental group (n = 10), a female control group (n = 10), and a female experimental group (n = 10); 1. Baseline test of defensive attack behavior: A plastic model snake equipped with a mechanical stimulation device and a mouse were placed in the same breeding cage. The crocodile clip clamped the mouse's tail to apply continuous harmful mechanical stimulation to the mouse. During this period, the latency of the mouse's first bite, the number of bites, and the total bite duration were recorded. The experiment lasted for 1 minute. After the experiment, the model snake was removed, disinfected with alcohol, and the mouse was returned to the original breeding cage; the experiment was repeated to test the defensive attack behavior of all mice, and the latency of their second bite, the number of bites, and the total bite duration were recorded.

[0028] 、Test after acute GLP-1RA exposure: Within 4 days after the baseline test, the defensive attack behavior of the mice was tested once a day for 4 consecutive days, as described below: On the second day after the baseline test, 20 minutes before the defensive attack behavior test, each mouse in the male experimental group and the female experimental group was intracerebroventricularly injected with a GLP-1 receptor agonist (Liraglutide, 0.5 μg / μL, ICV) at a dose of 2 μL / mouse; each mouse in the male control group and the female control group was intracerebroventricularly injected with physiological saline at a dose of 2 μL / mouse; Place a plastic model snake equipped with a mechanical stimulation device and a mouse in the same breeding cage. Use an alligator clip to clamp the mouse's tail and apply continuous harmful mechanical stimulation to the mouse. During this period, record the latency of the mouse's first bite, the number of bites, and the total biting duration. The experiment lasts for 1 minute. After the experiment, remove the model snake, disinfect it with alcohol, and return the mouse to its original breeding cage. Repeat the experiment to conduct defensive attack behavior tests on all mice, and record their latency of the second bite, the number of bites, and the total biting duration. Within 4 days (including the day) after GLP-1 receptor agonist exposure, conduct a defensive attack behavior test once a day to detect the action time of GLP-1 receptor agonist exposure on the mouse's defensive attack behavior.

[0029] The experimental results are as Figure 6 、 Figure 7 shown: (1) Acute GLP-1RA exposure can significantly increase the latency of the mouse's first attack and reduce the number of attacks and the attack time of the mouse.

[0030] (2) Acute GLP-1RA exposure shows different effects in male and female mice. In male mice, within two days after acute GLP-1RA exposure, the latency of the first attack and the number of attacks both change significantly. However, the change in the defensive attack behavior of female mice caused by acute GLP-1RA exposure is only effective on the day of exposure. It is proved that the influence time of GLP-1RA on the mouse's defensive attack behavior is related to gender to a certain extent, and its influence effect is more significant for male mice.

[0031] Example 3: Effect of acute GLP-1RA exposure on the defensive attack behavior of socially isolated mice 1. Social isolation Select the male mice in Preparation Example 1 (n = 42), and randomly divide them into a control group (n = 14), a blank group (n = 14), and an experimental group (n = 14). Keep the mice in the blank group and the experimental group separately for 3 weeks to simulate social isolation. Keep the mice in the control group in groups according to the original conditions for 3 weeks.

[0032] 2. Open Field Test (OFT): On the second day after the end of social isolation, each mouse in the experimental group was intracerebroventricularly injected with a GLP-1 receptor agonist (Liraglutide, 0.5 μg / μL, ICV) at a dose of 2 μL / mouse; each mouse in the blank group and the control group was intracerebroventricularly injected with normal saline at a dose of 2 μL / mouse. Twenty minutes after the drug injection, a mouse was placed in the central area of a transparent cubic open field of 40 cm × 40 cm × 40 cm and allowed to freely explore for 5 minutes. The total distance moved by the mouse was recorded. After the test, the mouse was returned to the breeding cage. After cleaning the open field with 75% alcohol, the next mouse was tested. The experiment was repeated for all mice, and the total distance moved by each mouse in the open field and the time spent in the central area were recorded.

[0033] The experimental results are as Figure 9 shown: Compared with group-housed mice, social isolation and GLP-1RA exposure had no effect on the total distance moved by mice. However, the percentage of time that socially isolated mice spent in the central area was significantly reduced compared with group-housed mice, indicating that socially isolated mice exhibited anxiety-like behaviors.

[0034] Defensive Attack Test (DAT) On the second day after the open field test and 20 minutes before the start of the defensive attack behavior test, the mice used in step 2 were continued to be used. Each mouse in the experimental group was intracerebroventricularly injected with a GLP-1 receptor agonist (Liraglutide, 0.5 μg / μL, ICV) at a dose of 2 μL / mouse; each mouse in the blank group and the control group was intracerebroventricularly injected with normal saline at a dose of 2 μL / mouse. A plastic model snake equipped with a mechanical stimulation device and a mouse were placed in the same breeding cage. The alligator clip was used to clamp the mouse's tail, and continuous harmful mechanical stimulation was applied to the mouse. During the experiment, the latency to first bite, the number of bites, and the total bite duration of the mouse were recorded. The experiment lasted for 1 minute. After the test, the model snake was removed, disinfected with alcohol, and the mouse was returned to the original breeding cage. The experiment was repeated for all mice, and the latency to first bite, the number of bites, and the total bite duration of each mouse were recorded.

[0035] The experimental results are as Figure 10 shown: (1) Compared with the control group mice, the latency to first attack of the blank group mice was significantly shortened, and at the same time, the number of attacks and the attack time were significantly increased. This result indicates that social isolation significantly increased the aggressiveness of mice.

[0036] (2) Compared with the blank group mice, the latency to first attack of the experimental group mice was significantly prolonged, and the number of attacks and the attack time were shortened, indicating that GLP-1RA can alleviate the increased aggressiveness induced by social isolation and has potential value for psychological stress intervention and can be used for the treatment of diseases such as social anxiety and aggression disorder.

[0037] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention. The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Use of a GLP-1 receptor agonist in the preparation of a drug for reducing defensive aggressive behavior.

2. The use according to claim 1, characterized in that: The GLP-1 receptor agonist includes exenatide, benaglutide, lixisenatide, liraglutide, dulaglutide, semaglutide, polyethylene glycol loxenatide, albiglutide or tilportide.

3. The use according to claim 1, characterized in that: The defensive aggressive behavior includes defensive aggressive behavior associated with mental illness and / or organic brain damage.

4. The use according to claim 1, characterized in that: The defensive aggressive behavior includes defensive aggressive behavior associated with post-traumatic stress disorder, schizophrenia, autism spectrum disorder, social anxiety disorder, borderline personality disorder, bipolar disorder, Alzheimer's disease and / or organic brain injury.

5. The use according to claim 1, characterized in that: The GLP-1 receptor agonist is administered orally, subcutaneously, intravenously and / or nasally.

6. The use according to claim 1, characterized in that: The drug is prepared into one of the following dosage forms: tablets, capsules, granules, powders, suspensions, emulsions, powders, solutions, gels, syrups, pills, tinctures, wine preparations, decoctions, lozenges, mixtures, suppositories, injections, inhalants or sprays.

7. The use according to claim 1, characterized in that: The medicament further comprises at least one pharmaceutically acceptable carrier or excipient.

Images