CN115137720A

CN115137720A - Application of citrulline in preparation of anxiolytic drugs

Info

Publication number: CN115137720A
Application number: CN202210892680.7A
Authority: CN
Inventors: 刘兴国; 杨亮; 阮子峰
Original assignee: Guangzhou Institute of Biomedicine and Health of CAS
Current assignee: Guangzhou Institute of Biomedicine and Health of CAS
Priority date: 2022-07-27
Filing date: 2022-07-27
Publication date: 2022-10-04

Abstract

The invention provides application of citrulline or pharmaceutically acceptable salt thereof in preparing medicines and a pharmaceutical composition for preventing or treating anxiety disorder, and the citrulline or pharmaceutically acceptable salt thereof has important research value and clinical application value for preventing or treating the anxiety disorder.

Description

Application of citrulline in preparation of anxiolytic drugs

Technical Field

The present invention relates to the field of medicine. In particular, the invention relates to application of citrulline in preparing an anxiolytic drug.

Background

Anxiety disorders and anxiety symptoms associated with psychiatric disorders are a group of neuropsychiatric disorders in which paroxysmal or persistent anxiety and stress are the main features. The incidence of anxiety disorders has increased year by year in recent years, severely affecting the quality of life of patients. At present, drugs and psychological treatments are mainly used for treating anxiety, but drugs for treating anxiety still need to be researched.

Disclosure of Invention

The present invention aims to solve, at least to some extent, the technical problems of the prior art. Therefore, the invention provides the application of citrulline or a pharmaceutically acceptable salt thereof in preparing medicines and a pharmaceutical composition for preventing or treating anxiety neurosis, and the citrulline or the pharmaceutically acceptable salt thereof has important research value and clinical application value for preventing or treating the anxiety neurosis.

In one aspect of the invention, the invention proposes the use of citrulline, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament. According to an embodiment of the invention, the medicament is for the prevention or treatment of anxiety disorders.

In still another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating anxiety disorders. According to an embodiment of the invention, the pharmaceutical composition comprises citrulline or a pharmaceutically acceptable salt thereof.

Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.

Drawings

The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:

FIG. 1 shows open field experiments of wild type and mtDNA mutant mice according to example 1 of the present invention, and statistics of mean velocity and frequency measurements of mice entering the central region (n.gtoreq.3,. P.lt0.05, independent t-test);

FIG. 2 shows a small intestine metabolite detection map of wild type and mtDNA mutant mice according to example 1 of the present invention (n.gtoreq.4,. Gtoreq.p < 0.05, independent t-test);

FIG. 3 shows open field experiments after administration of citrulline to mtDNA mutant mice according to example 2 of the present invention, and statistics of mean velocity and frequency of entry into the central region of mtDNA mutant mice (n =2,. Sup.p < 0.05, independent t-test).

Detailed Description

The following describes embodiments of the present invention in detail. The following examples are illustrative only and are not to be construed as limiting the invention.

The present invention provides the use of citrulline or a pharmaceutically acceptable salt thereof for the preparation of a medicament and a pharmaceutical composition for the prevention or treatment of anxiety, which will be described in detail, respectively, below.

Application of citrulline or pharmaceutically acceptable salt thereof in preparation of medicines

In one aspect of the invention, the invention proposes the use of citrulline or a pharmaceutically acceptable salt thereof in the manufacture of a medicament. According to an embodiment of the invention, the medicament is for the prevention or treatment of anxiety disorders.

The POLG mutant mouse has a D257A mutation at the position of POLG exonuclease so that the exonuclease activity of the POLG mutant mouse is lost, correction defects in the mtDNA replication process are caused, mtDNA mutation is caused, and the POLG mutant mouse is a well-known mouse model for researching mtDNA mutation and can generate anxiety symptoms. Further, the inventors conducted metabolomics studies on the small intestine of mtDNA mutant mouse models, and found that the small intestine citrulline specificity was decreased in the mouse models. Further, by administering citrulline to mtDNA mutant mouse model, the symptoms of anxiety in mice can be effectively alleviated. Therefore, the citrulline or the pharmaceutically acceptable salt thereof can prevent or treat the anxiety disorder, and has important significance for the research and clinical application of the anxiety disorder.

The term "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastrointestinal upset, dizziness and the like, when administered to a human. Preferably, the term "pharmaceutically acceptable" as used herein refers to those approved by a federal regulatory agency or a state government or listed in the U.S. pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

According to an embodiment of the invention, the drug is targeted to the brain-gut axis. The brain-gut axis (brain-gut axis) refers to a two-way information communication network between the brain and the gut, and broadly includes neural interaction pathways, neuroendocrine and neuroimmune pathways, gut microbiota, and the like. The inventor carries out detection analysis on mouse metabolites with low immunity and finds that the citrulline in the small intestine is reduced, and the small intestine is directly related to the brain intestinal axis. After the citrulline is given to the mice, the low immunity of the mice is relieved, so that the action target point of the citrulline is shown to be the brain-intestine axis.

According to an embodiment of the present invention, the administration mode of the drug is oral or enema. As described above, citrulline can prevent or treat anxiety disorders by targeting the brain-intestine axis, and thus, it can act on the brain-intestine axis to exert the drug effect better by oral administration or enema administration.

The administration frequency and dose of the drug of the present invention can be determined by a number of relevant factors, including the type of disease to be treated, the administration route, the age, sex, body weight and severity of the disease of the patient, and the type of drug as an active ingredient. According to some embodiments of the invention, the daily dose may be divided into 1, 2 or more doses in a suitable form for administration 1, 2 or more times over the entire period, as long as a therapeutically effective amount is achieved.

Pharmaceutical composition for preventing or treating anxiety disorders

In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating anxiety disorders. According to an embodiment of the invention, the pharmaceutical composition comprises citrulline or a pharmaceutically acceptable salt thereof.

According to an embodiment of the invention, the pharmaceutical composition further comprises: pharmaceutically acceptable adjuvants.

The invention does not strictly limit the types of the auxiliary materials and can flexibly select the auxiliary materials according to the conditions. For injectable formulations, pharmaceutically acceptable carriers may include buffers, preservatives, analgesics, solubilizers, isotonic agents (isotonicagents) and stabilizers. For formulations for topical administration, pharmaceutically acceptable carriers may include bases, excipients, lubricants, and preservatives. The pharmaceutical composition of the present invention may be prepared in various dosage forms in combination with the above pharmaceutically acceptable carrier. For injectable preparations, the pharmaceutical compositions may be prepared in ampoules, e.g. in single dose dosage form, or in unit dosage forms, e.g. in multidose containers. The pharmaceutical compositions may also be formulated as solutions, suspensions, tablets, pills, capsules and depot preparations. Among the excipients and diluents suitable for pharmaceutical formulations according to some embodiments of the present invention may be, among others: lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. According to other embodiments of the present invention, the adjuvants of the present invention may further comprise fillers, anticoagulants, lubricants, moisturizers, fragrances, and preservatives.

The scheme of the invention will be explained with reference to the following examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

Example 1

1. Wild mice and mtDNA mutant mice of 8 months old are selected and transferred to a laboratory of an open field experiment, so that the mice adapt to a new environment. One day after acclimation, the mice were subjected to open field experiments. Mice were first placed in the center of a 50 × 50 × 50cm (length × width × height) chamber for 10 minutes, recorded with a camera positioned above the square box, and manipulated with the nouldsi ethovision 9.0 software. Between tests, the floor was cleaned with 75% alcohol. During the open field experiments, all behaviors such as average speed and frequency of entry into the central zone were recorded and analyzed to assess anxiety levels in 8-month old wild-type and mtDNA mutant mice.

As shown in fig. 1, the frequency and average velocity of entry of mtDNA mice into the central zone were less as a result of the open field experiment, indicating increased anxiety in mtDNA mutant mice. These results indicate that the function of the gut axis is affected during anxiety.

2. In order to search for a potential mechanism for regulating the brain-intestinal axis function and detect the small intestine metabolomics of mtDNA mutant mice, the method comprises the following specific steps:

(1) Taking small intestine, taking small intestine and middle intestine sections of about 3-5cm, washing chyme in the intestine sections by using cold PBS (phosphate buffer solution), paying attention to washing strength to avoid damaging small intestine structures, and quickly freezing by using liquid nitrogen. Taking out the sample from a refrigerator at the temperature of-80 ℃, unfreezing the sample on ice, weighing 50mg of the sample in a 2mL EP tube after unfreezing, and adding 500uL of 70% methanol water internal standard extracting solution precooled at the temperature of-20 ℃.

(2) One small steel ball was added and homogenized at 30Hz for 4 times, 30s each time.

(3) After homogenization, oscillating at 1500r/min for 5min, and standing on ice for 15min.

(4) Centrifugation was carried out at 10min,4 ℃ and 12000r/min.

(5) 200uL of the supernatant was taken and loaded into the corresponding vial liner for LC-MS/MS analysis.

The results are shown in fig. 2, the specificity of small intestine citrulline in mtDNA mutant mice is reduced, suggesting that citrulline plays an important role in the brain-intestine axis.

Example 2

200mg L-citrulline is weighed and added to 250mLSPF grade animal drinking water, mixed well and dissolved. The 8-month-old mtDNA mutant mice were weighed and divided equally into two groups by weight, one group was 250 mlsff-grade animal drinking water (control group) and the other group was 250 mlsff-grade animal drinking water containing 200mg L-type citrulline (experimental group). Water was supplied continuously for 14 days. After 14 days without intermittent water supply, the anxiety level of the mice was measured by open field experiments.

As shown in fig. 3, the frequency and average velocity of mtDNA mutant mice entering the central region increased after citrulline was added as a result of the open field experiment, indicating that there was less anxiety, indicating that citrulline alleviated the anxiety of mtDNA mutant mice.

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Moreover, various embodiments or examples and features of various embodiments or examples described in this specification can be combined and combined by one skilled in the art without being mutually inconsistent.

Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.

Claims

1. Use of citrulline or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of anxiety.

2. The use according to claim 1, wherein the drug is targeted to the brain-gut axis.

3. The use according to claim 1, wherein the medicament is administered orally or as an enema.

4. A pharmaceutical composition for preventing or treating anxiety disorders, comprising citrulline or a pharmaceutically acceptable salt thereof.

5. The pharmaceutical composition of claim 4, further comprising: pharmaceutically acceptable adjuvants.