WO2020088270A1 - Application of kynurenic acid or derivative thereof in preparing drug for improving pathological damages associated with chronic psychological stress - Google Patents

Abstract

An application of kynurenic acid or derivative thereof in preparing a drug for improving pathological damages associated with chronic psychological stress. Compared with the prior art, disclosed is an application of kynurenic acid or derivative thereof in preparing a drug for improving pathological damages associated with chronic psychological stress. Related researches prove that oral administration of kynurenic acid has a good effect in improving impaired social interaction, inflammatory disorders of the central nervous system, and splenic immune dysregulation associated with chronic psychological stress.

Description

Application of kynurenine quinolinic acid or its derivatives in preparation of drugs for improving chronic mental stress related pathological damage Technical field

The invention discloses the application of canine urinary quinolinic acid or its derivatives in the preparation of drugs for improving pathological damage related to chronic mental stress, and belongs to the technical field of new uses of compounds.

Background technique

Chronic psychological stress is common among people in modern society, and is found to be closely related to the increase or recurrence of various diseases (such as psychosis, coronary heart disease, and inflammatory bowel disease). Studies have shown that chronic mental and psychological stress can cause central and peripheral endocrine-inflammatory-immune reaction disorders, accompanied by systemic abnormal symptoms such as depressed mood, reduced learning and memory ability, and gastrointestinal dysfunction. With the further acceleration of life rhythm, the social and economic burden associated with chronic social psychological pressure will be further intensified, and finding effective intervention methods has become an urgent scientific and social problem to be solved.

More and more evidences show that the changes of endogenous small molecule metabolites under chronic psychological stress are involved in the occurrence and development of multiple organ dysfunction. By restoring the balance of metabolic pathways or its network, it can help to improve multiple pathological links such as abnormal mood, vascular endothelial damage, and intestinal inflammation, thereby effectively resisting the health damage caused by chronic mental stress. Therefore, the regulation strategy to correct endogenous small molecule metabolic imbalance has potential application value in improving the pathological process related to chronic mental stress.

Kynurenic acid is a metabolite of kynurenine under the action of kynurenine aminotransferase (KAT) in the body. It is reported in the literature that it can act on the N-methyl-D-aspartate receptor (N-methyl-D-aspartate receptor, NMDAR), α7 nicotinic cholinergic receptor (α7nicotine cholinergic subtype alpha 7receptor, α7nAChR) and G-protein coupled receptor (G-protein coupled receptor 35, GPR35), with Certain nerve activity and immune regulation. Current basic and clinical studies have shown that canine urinary quinolinic acid levels are abnormal in a variety of central disease states. An important in vivo process characteristic of kynurenic acid is its poor permeability to the physiological barrier, and kynurenic acid in the peripheral circulation cannot pass the blood-brain barrier. Based on the traditional understanding that drugs must be efficiently distributed in the brain to regulate the central pathological process, the current reports are focused on adjusting the level in the brain and regulating cognition by intracerebral injection or increasing the precursor kynurenine level. Obstacles and other diseases.

Summary of the invention

Object of the invention: The purpose of the present invention is to disclose the application of kynuria quinolinic acid or its derivatives in the preparation of drugs for improving chronic mental stress-related pathological damage. The role of chronic mental stress-related social disorders, central inflammation disorders, and spleen immune disorders.

Technical solution: In order to achieve the above-mentioned object of the invention, the present invention adopts the following technical solution:

The application of kynurenine quinolinic acid or its derivatives in the preparation of drugs for improving pathological injuries related to chronic mental stress.

The chemical name of the canine urinary quinoline acid is 4-hydroxyquinoline-2-carbo xylic acid, the molecular formula is C ₁₀ H ₇ NO ₃ , the molecular weight is 189.17g / mol, and its chemical structural formula as follows:

Preferred:

Application of the kynurenine quinolinic acid or its derivatives in the preparation of drugs for treating depression.

The medicament for treating depression can simultaneously improve depressive behavior and accompanying central-peripheral immune abnormalities.

Depressive behaviors such as chronic mental stress-induced social disorders and other depressive behaviors; the central-peripheral immune abnormalities are inflammatory cytokine levels and inflammatory cell infiltration disorders accompanying brain depression and spleen sites.

The improvement of chronic mental stress-related pathological damage refers to the improvement of chronic mental stress-induced social disorders and immune disorders in brain, spleen and other parts.

The medicament is a medicament made from quinolonic acid as a single component.

Alternatively, the medicine is prepared with kynurenine quinolinic acid or its derivatives as active ingredients, plus pharmaceutically acceptable auxiliary materials.

The dosage form of the medicine is ordinary tablet, ordinary capsule, oral solution, syrup, granule, dripping pill, oral disintegrating tablet, sustained release tablet, controlled release tablet, sustained release capsule, controlled release capsule, injection or infusion Agent.

The pharmaceutically acceptable auxiliary materials in the present invention refer to the addition of various conventional auxiliary materials required when preparing different dosage forms, such as diluents, binders, disintegrants, glidants, lubricants, flavoring agents, inclusions The materials, adsorption materials, etc. are prepared into any commonly used preparations by conventional preparation methods.

Technical effect: Compared with the prior art, the present invention discloses the application of canine urinary quinolinic acid or its derivatives in the preparation of drugs for improving pathological damage related to chronic mental stress. The present invention has confirmed through relevant studies that canine urinary quinolinic acid has It can effectively improve social disorders related to chronic mental stress, central inflammation disorders, and immune dysregulation of spleen.

BRIEF DESCRIPTION

Figure 1: The effect of canine urea quinolinic acid on the social avoidance behavior of chronic social stress mice. Control is a normal control group mouse, CSDS is a chronic social stress group mouse, and CSDS + KA is given during chronic social stress. For KA mice, CSDS was given only blank vehicle; the results showed that KA can significantly increase the residence time of mice in the central social area (Figure A, #p <0.05, compared with the CSDS group) and frequency (Figure B, #p <0.05 , Compared with the CSDS group).

Figure 2: The effect of canine urinary quinolinic acid on the infiltration of proinflammatory monocytes in the brain of chronic social stress mice. Control is a normal control group mouse, CSDS is a chronic social stress group mouse, and CSDS + KA is chronic KA mice were given social stress, and CSDS was given only blank vehicle; the results showed that KA can significantly reduce the proinflammatory monocyte infiltration in the hippocampus Ly6C ^hi (Figure A, CD11b ⁺ Ly6C ^hi cell grouping diagram; Figure B, Ly6C ^hi Statistical graph of the proportion of proinflammatory monocytes, * p <0.05).

Figure 3: The effect of kynuria quinolinic acid on chronic social stress-induced central inflammatory mediators in mice. Control is a normal control group, CSDS is a chronic social stress group, and CSDS + KA is a chronic social stress. KA mice were given in the middle, and CSDS was only given a blank vehicle; the results showed that KA can significantly reduce the inflammatory factor Ccl2 in the hippocampus (Figure A, #p <0.05, compared with the CSDS group) and Il6 (Figure B, #p <0.05, compared with the CSDS Group comparison) mRNA levels.

Figure 4: The effect of canine urinary quinolinic acid on chronic social stress-induced spleen immune cell infiltration in mice. Control is the normal control group, CSDS is the chronic social stress group, and CSDS + KA is the chronic social stress. KA mice were given in the middle, and CSDS was only given a blank vehicle; the results showed that KA had no significant effect on the proportion of CD4 ⁺ T cells (Figure A), but could significantly increase CD8 ⁺ T cells in the spleen of chronic social stress mice (Figure B, * p <0.05) and reduce the proportion of NK1.1 cells (Figure B, *** p <0.001).

detailed description

The present invention will be further described in detail below in conjunction with specific embodiments.

Example 1: Regulatory effect of canine urinary quinolinic acid on social social avoidance behavior induced by chronic social stress in mice

(1) Materials: C57BL / 6J mice (male, 8 weeks old), after acclimating to the environment for 1 week, were randomly divided into three groups, namely normal control group (Control group, n = 6), chronic social stress model group (CSDS group, n = 8) and canine urine quinolinic acid intervention group (CSDS + KA group, n = 8). Place the stressed mice on the side of the attacking mice at 16:00 pm every day, accept the 10-minute challenge, and then transfer them to the other side of the same cage for 24 hours; repeat the above operation once a day for 10 days; The normal control mice were fed in a single cage without receiving social stress. Mice in the intervention group were given quinolinic acid (20 mg / kg, adjusted to pH with 1M NaOH and dissolved in pure water) by intragastric administration 0.5 hours before daily stress. Social experiment evaluation was conducted on the 11th day.

(2) Social avoidance experiment: The experiment is divided into two stages. In the first stage, the empty perforated mouse cage is placed in the social area of the open field, and the mouse to be tested is placed to allow it to walk freely in the social open field. Record its trajectory within 150 seconds and the time it stays in the social area, and then remove the mouse and the perforated mouse cage, clean and wipe the perforated mouse cage with the attacking mouse back into the social area (same location) , Put the same C57 mouse into the social open field again, record its movement trajectory within 150 seconds and the time it stays in the social area, compare the difference between the movement trajectories of the mice within 150 seconds, and calculate the social area (interaction zone) ) Residence time and times.

(3) Data analysis method: The experimental data is expressed in Mean ± SD, and the results are analyzed and processed using GraphPadPrism6 (GraphPadSoftwareInc.) Software. The data is analyzed by One-way ANOVA, p <0.05 indicates that the data difference has statistics significance.

Example 2: Regulatory effect of canine urinary quinolinic acid on chronic social stress-induced central inflammation disorder in mice

(1) Materials: The specific method is the same as in Example 1.

(2) Isolation of brain immune cells and flow cytometry to examine the infiltration of CD45 ^hi CD11b ⁺ Ly6C ^hi monocytes in the mouse brain: after the mice were sacrificed, the blood was removed by perfusing 20 mL of ice PBS solution through the left ventricle until The liver is whitish, and the brain tissue is carefully separated, removed and placed in ice-free anti-RPMI1640 medium. Then transfer to a 5mL glass homogenizer, add 3mL ice PBS solution to each brain tissue for homogenization, make up the homogenate to 7mL and mix with 100% Percoll solution to make a 30% Percoll cell suspension (10mL) At a constant speed, slowly add it to 2mL of 70% Percoll separation solution, centrifuge at 500g for 30min at 18 ° C, and slowly and carefully remove the turbid layer at the interface between 30% and 70% Percoll with a pipette to obtain lymphocyte population. After washing twice with PBS solution, it was resuspended in staining buffer to 100 μL of cell suspension for flow antibody incubation. 1 μL of anti-CD16 / 32 antibody was added to 100 μL of cell suspension and blocked in the ice bath for 10 min in the dark. Add the corresponding specific flow cytometry antibodies (anti-CD45-PE, anti-CD11b-APC, anti-Ly6C-FITC) without washing, incubate for 45min in an ice bath protected from light, and then wash three times with ice PBS solution (at 4 ° C) Centrifuge at 400g for 5min), finally resuspend and fix with 4% paraformaldehyde, and test it on the next day. Select the appropriate channel for flow analysis, and the related flow data is analyzed by FlowJo software.

(3) PCR analysis of the inflammatory factor mRNA in the hippocampus: remove the unilateral hippocampus of the mouse into a centrifuge tube, add 1.0m L Trizol and then homogenize with ice bath ultrasound, add 250 μL of chloroform after standing, mix upside down for 20s, After fully emulsified, let stand for 10min; centrifuge at 12,000g for 15min at 4 ℃, transfer the colorless supernatant liquid (300μL) to a new centrifuge tube; add equal volume of isopropanol, continue to invert and mix for 20s, let stand 10min; centrifuge at 12,000g for 10min at 4 ° C; carefully pour out the supernatant, leaving the precipitate at the bottom of the centrifuge tube; slowly add 1mL of ice 75% ethanol (RNase-free water preparation), wash the bottom precipitate upside down; at 4 ° C Centrifuge at 12,000g for 5 minutes, pour out the supernatant, and leave it at room temperature for 10 minutes to wait for the 75% ethanol solution to evaporate. Dissolve the bottom precipitate with 10 μL RNase-free water. After completely dissolved, it can be stored at -80 ° C or measured for concentration. Reverse transcribe 500ng mRNA into cDNA according to the instructions of the reverse transcription kit, and then operate according to the requirements of the SYBR GREEN real-time quantitative PCR instruction manual. Using Gapdh as the reference gene, the 2- ^ΔΔCT method was used to semi-quantitatively analyze the relative difference in the expression of the target gene in different samples.

Example 3: Regulatory effect of canine urinary quinolinic acid on chronic social stress-induced spleen immune disorder in mice

(1) Materials: The specific method is the same as in Example 1.

(2) Isolation of splenic immune cells and flow cytometry: Place the mouse spleen on a 70μm cell screen, add 2mL of PBS solution, squeeze and grind the spleen into a cell suspension using a 5mL syringe piston, and slowly add to Lympholyte-M On the mononuclear cell separation solution (2mL, v / v 1: 1), centrifuge at 1200g for 20min at room temperature, and slowly remove the intermediate white turbid cells with a pipette to obtain a lymphocyte population. Wash twice with PBS solution. Red blood cell lysate can be used to remove residual red blood cells. Resuspend the flow suspension into 100μL cell suspension for flow antibody incubation. The method is the same as in Example 2.

Claims (8)

1. The application of kynurenine quinolinic acid or its derivatives in the preparation of drugs for improving pathological injuries related to chronic mental stress.
2. The use according to claim 1, characterized in that the kynurenine quinolinic acid or its derivative is used in the preparation of a medicament for treating depression.
3. The use according to claim 2, wherein the drug for treating depression can simultaneously improve depressive behavior and accompanying central-peripheral immune abnormalities.
4. The use according to claim 3, characterized in that the depressive behavior is depressive behavior such as chronic mental stress-induced social disorder; the central-peripheral immune abnormality is the level of inflammatory factors in the brain and spleen associated with the depressive behavior And inflammatory cell infiltration disorder.
5. The use according to claim 1, wherein the improvement of chronic mental stress-related pathological damage refers to improvement of chronic mental stress-induced social disorders and immune disorders in the brain and spleen.
6. The use according to claim 1, characterized in that the drug is made of kynurenine or its derivatives as a single component.
7. The use according to claim 1, characterized in that the medicine is prepared by using kynuria quinolinic acid or its derivatives as the active ingredient, plus pharmaceutically acceptable auxiliary materials.
8. The use according to claim 1, characterized in that the dosage form of the medicine is ordinary tablets, ordinary capsules, oral liquid, syrup, granules, dropping pills, oral disintegrating tablets, sustained release tablets, controlled release tablets, Slow-release capsules, controlled-release capsules, injections or infusions.

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