CN110038017A - The new application of 5-aminosalicylic acid - Google Patents

Abstract

The invention discloses 5-aminosalicylic acids to treat the purposes in drug of the depression with anxiety state in preparation.5-aminosalicylic acid of the present invention can well alleviate depression with anxiety state, and reduce its hippocampus level of inflammation, help to improve anti depressant medication effect, reduce drug side-effect.

Description

New uses of 5-aminosalicylic acid

technical field

The present invention relates to a new use of 5-aminosalicylic acid, in particular to the use in the preparation of a medicament for treating depression accompanied by anxiety.

Background technique

Depression is a common mood disorder, and its core symptoms are low mood, slow thinking, decreased volitional activity, sleep disturbance, and repeated suicidal thoughts. According to WHO statistics, there are currently about 121 million depression patients in the world, and 850.000 deaths are related to depression every year.

Depression has many accompanying symptoms, and anxiety symptoms are one of the main accompanying symptoms of depression, and its incidence can be as high as 75%. The pathogenesis of anxiety state in depression is complex, and its mechanism is still unclear. SSRIs or SNRIs are commonly used in clinical depression treatment, but they are less effective for anxiety symptoms in patients with depression. Therefore, finding a drug that can effectively improve the anxiety symptoms of depression has practical significance for improving the therapeutic effect of depression and reducing the dosage to reduce its side effects.

5-aminosalicylic acid (mesalazine, 5-ASA), its structural formula is It is a drug used to treat ulcerative colitis and Crohn's disease. There are currently no reports of its efficacy in treating anxiety symptoms of depression.

SUMMARY OF THE INVENTION

The object of the present invention is to provide new uses of 5-aminosalicylic acid.

The invention provides the use of 5-aminosalicylic acid in preparing a medicine for treating anxiolytic symptoms.

Wherein, the drug is a drug for treating anxiety symptoms of depression.

Wherein, the drug is a drug for treating depression accompanied by anxiety state caused by intestinal microbial disturbance.

Wherein, the drug is a drug for inhibiting hippocampal inflammation.

The invention provides the use of 5-aminosalicylic acid in preparing a medicament for treating hippocampal inflammation.

The invention also provides a medicine for treating depression accompanied by anxiety state, which is a preparation prepared by using 5-aminosalicylic acid as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary components.

Wherein, the formulation is an oral formulation.

The experiment of the present invention confirms that oral administration of 5-aminosalicylic acid can anxiolytic, significantly improves the hippocampal inflammatory state and the anxiety state of mice accompanied by anxiety and depression, indicating that 5-aminosalicylic acid can be used for the treatment of depression accompanied by anxiety It provides a new option for clinical treatment of depression accompanied by anxiety state.

Obviously, according to the above-mentioned content of the present invention, according to the common technical knowledge and conventional means in the field, without departing from the above-mentioned basic technical idea of the present invention, other various forms of modification, replacement or change can also be made.

The above content of the present invention will be further described in detail below through the specific implementation in the form of examples. However, this should not be construed as limiting the scope of the above-mentioned subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention belong to the scope of the present invention.

Description of drawings

Figure 1 Behavioral performance of mice after fecal transplantation. Depressed patient fecal transplant mice showed higher levels of anxiety and depression than normal human fecal transplant mice.

Figure 2. Behavioral performance of fecal transplant mice from depressed patients after using 5-aminosalicylic acid. Fecal transplanted mice from depressed patients showed reduced anxiety levels after administration of 5-aminosalicylic acid.

Figure 3. The expression levels of inflammatory factors and their related receptors in the hippocampus of the mice in Figures 1 and 2 obtained by qRT-PCR. 5-aminosalicylic acid can completely reverse the expression of TLR4, an inflammation-related receptor in the hippocampus of mice induced by gut microbes in depressed patients, and restore the anti-inflammatory factor IL6 to normal levels.

Detailed ways

Example 1 Experiment of 5-aminosalicylic acid of the present invention in the treatment of depression accompanied by anxiety

1. Model preparation of depression accompanied by anxiety

Stool samples containing gut microbes for transplantation were randomly selected from clinical male patients with major depression and healthy male controls. Fecal treatment was carried out in a sterile environment, 0.1 g of each fecal specimen was suspended in 1.5 ml of sterile PBS, and 5 equal parts of the fecal suspension were mixed together. Adult male Kunming mice (15-30 mice) (6-8 weeks) were colonized by gavage of the suspension (0.2 ml, 1 gavage) via a gavage needle. The mixed fecal suspensions of severe depression or healthy controls were introduced into different isolators through sterile transfer windows, and all colonization operations were carried out in the isolators, which were strictly sterile.

Behavioral testing was performed the second week after fecal colonization. The fecal transplant mice of depressed patients and normal human fecal transplant mice were analyzed by tail suspension experiment, forced swimming experiment and field experiment to determine whether the model was successful:

Field experiment:

Field experiments are used to assess motor ability, as well as anxiety-related behaviors. The behavioral testing instrument is a square box made of black plexiglass without a lid, with dimensions of 45cm×45cm×30cm (length×width×height). Each mouse was placed individually in a fixed corner of the box, allowing the mice to freely explore the box for 6 minutes. The entire behavior of the mice was recorded by a video camera, and the last 5 minutes of the video were analyzed using the video trajectory analysis software (SMART, Spain/Panlab). The total distance exercised reflects the exercise capacity of mice. The percentage of time spent moving in the middle quarter of the box at the bottom of the box is called the central movement time. as a criterion for anxiety-like behavior.

Tail suspension experiment:

The experimental apparatus for the tail suspension experiment was a black Plexiglas box with a hook on the top. At the beginning of the experiment, the mice were wrapped with scotch tape 2 cm from the tip of the tail and hung upside down on the top of the box. The whole process of the experiment was recorded by a camera. The experimental process lasted for 6 minutes, and the immobility time was analyzed for the last 5 minutes. If the experimental mouse climbed on the hook, the mouse was excluded from the experiment. When the mouse body is still and passively suspended, it is regarded as immobility time.

Forced Swimming:

The device for forced swimming is a cylindrical, lidless container made of transparent plexiglass and has a size of 15 cm in diameter and 30 cm in height. Fill the container with warm water with a depth of about 20cm and a temperature of 23-25 degrees. The mice were put into the apparatus one by one, the camera was aligned horizontally in the container, and the behavior of the mice within 6 minutes of entering the water was recorded. The last 5 minutes of the video were analyzed using video trajectory analysis software (SMART, Spain/Panlab) and the resting time was recorded. Immobility time responds to depression-like behavior in mice.

The experimental results are shown in Figure 1. The immobility time of fecal transplanted mice from depression patients was significantly higher than that of normal human fecal transplanted mice, and the central movement distance was significantly lower than that of normal human fecal transplanted mice. Anxiety symptoms and depression state, indicating that the model of depression anxiety symptoms model was successfully established, which can be used to screen corresponding therapeutic drugs.

2. 5-aminosalicylic acid of the present invention relieves depression and anxiety symptoms

First, on the basis of the depression anxiety symptom model established above, 5-aminosalicylic acid was introduced into different sterile isolators through a sterile transfer window through a sterile centrifuge tube, and 5-aminosalicylic acid was transferred into the isolator. Sylic acid and sterilized drinking water were prepared at a ratio of 5 mg/ml, and the aforementioned depression and anxiety symptom model mice were allowed to freely ingest, and the dressing was changed every other day. All experiments were carried out in an isolator, strictly sterile, and the control group was given Sterilized drinking water.

Behavioral testing was performed the second week after dosing. The effect of 5-aminosalicylic acid was detected by the elevated plus maze and the new environment feeding inhibition experiment:

Novelty suppressed feeding test

The mice were fasted 24 hours before the start of the experiment, while ensuring that the mice had an adequate supply of drinking water. Thirty minutes before the experiment, mice were acclimated by transferring them to clean cages. The experimental equipment is a black field box with a size of 45cm × 45cm × 30cm (length × width × height). A filter paper with a diameter of 12 cm is placed in the middle of the bottom of the field box, and a short mouse feed is placed in the middle of the filter paper. . At the beginning of the experiment, the mice were put in from a fixed corner of the open field, and the latency time was recorded from the time the mice were put in to pick up the food with their forelimbs. The length of time that mice eat reflects the degree of anxiety and depression in mice. The mice were then returned to their cages, and the food intake of the mice within 5 minutes was recorded to exclude the effect of differences in appetite on the timing of feeding.

Elevated plus maze

The elevated plus maze consists of two opposing open arms (30 cm x 5 cm) and two opposing closed arms (30 cm x 5 cm) criss-cross. Mice were placed in the middle of the junction, facing the open arm, and the trajectories were recorded for 5 minutes after the mice were acclimated to the maze for 1 minute. The time spent exploring the open arm and the time spent exploring the closed arm were recorded as indicators to evaluate mouse anxiety. After each mouse was tested, the maze was scrubbed with 10% alcohol to avoid the effect of odor and mouse residue on subsequent tests.

Fluorescence quantitative PCR (qRT-PCR)

Fresh frozen hippocampal RNA was extracted by grinding on ice using Trizol (Invitrogen, CA, USA). RNA was reverse transcribed into cDNA using PrimeScript RT Reagent Kit (Takara, Dalian, China). qRT-PCR was performed using LightCycler 96system (Roche, Germany), and the reagents were FastStart EssentialDNA Green Master (Roche), and the operation was performed according to the manufacturer's recommended method. Primers were screened from the validated Primerbank (Harvard) database and synthesized by Sangon Biotech (Shanghai, China). The target mRNA expression was corrected with β-actin and normalized with the control group.

Compared with the control group, the food intake of the 5-aminosalicylic acid group was significantly increased, the time of arm exploration was significantly shortened, and both male and female individuals had significant effects (as shown in Figure 2). Good anxiolytic effect.

As shown in Figure 3, the animal model hippocampus after 5-aminosalicylic acid treatment found that 5-aminosalicylic acid could completely reverse the expression of inflammation-related receptor TLR4 in the hippocampus of mice induced by gut microbes in depressed patients, and make The anti-inflammatory factor IL6 returned to normal levels, indicating that 5-aminosalicylic acid could exert anti-inflammatory effects in the hippocampus.

The 5-aminosalicylic acid of the invention can well relieve the anxiety symptoms of depression, and reduce the level of inflammation in the hippocampus, thereby helping to improve the therapeutic effect of antidepressants, reduce the side effects of the drugs, and improve the remission rate after the treatment of depression.

Claims (7)

1. Use of 5-aminosalicylic acid in the preparation of a medicament for treating anxiolytic symptoms. 2. The use according to claim 1, wherein the medicine is a medicine for the treatment of anxiety symptoms of depression. 3. The use according to claim 1 or 2, wherein the drug is a drug for inhibiting hippocampal inflammation. 4. Use of 5-aminosalicylic acid in the preparation of a medicament for inhibiting hippocampal inflammation. 5. The use according to claim 4, wherein the medicine is a medicine for treating hippocampal inflammation of depression and anxiety symptoms. 6. A medicine for the treatment of depression accompanied by anxiety state, characterized in that: it is a preparation prepared by using 5-aminosalicylic acid as an active ingredient, and adding pharmaceutically acceptable adjuvants or auxiliary ingredients. 7. The medicine according to claim 6, wherein the preparation is an oral preparation.