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Novel use of cardiocyte M3 receptor and IkM3 in screening medicament for treating arrhythmia

A technology for cardiac arrhythmia and cardiomyocytes, applied in the field of screening antiarrhythmic drugs, can solve problems such as side effects, proarrhythmia, and unknown functional uses of the heart

Active Publication Date: 2007-03-21
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some antiarrhythmic drugs have improved the patient's condition, there are still obvious shortcomings: ① The total effective rate of antiarrhythmic drugs is not high, only 30% to 60%; ② Ineffective for some stubborn arrhythmias; ③ Long-term The application has a poor effect on improving the survival rate of patients; ④ prone to problems such as arrhythmia-causing side effects
Further research found that M 3 The receptor has physiological functions such as hyperpolarizing the membrane potential, shortening the duration of the action potential, and slowing down the heart rate (Liu Yan, Wang Yue, Ma Manling, Zhang Yan, Li Houwei, Chen Qingwen, Yang Baofeng M3 receptor agonists on rat and rabbit hearts The influence of hemodynamics Acta Pharmaceutica Sinica, 2001; 36(2): 84-87), more importantly, the discovery of cardiac M 3 The receptor is closely related to arrhythmia and cardiomyocyte apoptosis (Liu Yan, Jing Yuhong, Sun Hongli, Li Hulun, Yang Baofeng, M 3 The relationship between the receptor and rat ischemic cardiomyocyte apoptosis (Journal of Pharmaceutical Sciences 2004, 39(5); 338-341), but so far, it is not known that cardiac M 3 Receptor and IK M3 functional use of

Method used

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Experimental program
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Effect test

specific Embodiment approach 1

[0005] Specific embodiment one: the cardiomyocyte M described in this embodiment 3 Receptor and IK M3 It has the application of screening drugs for treating arrhythmia. Add the drug to be screened in the extracellular fluid, and record the current intensity in the whole cell mode. If the current intensity is lower than that before the drug under the voltage of 50mv, it is a drug with curative effect for treating arrhythmia. The aforementioned screening method can be used to screen for inhibitory effect. medicine.

specific Embodiment approach 2

[0006] Specific implementation mode 2: This implementation mode utilizes cardiomyocyte M 3 Receptor and IK M3 The method for screening drugs for the treatment of arrhythmia comprises the following steps: a. isolating a single cardiomyocyte; b. establishing a whole-cell patch clamp detection mode; c. adding M to the extracellular fluid 3 Receptor agonist; d. Add other ion channel blockers to the extracellular fluid; e. Add drugs of different solubility to the extracellular fluid, record the current intensity in the whole cell mode, and the current intensity at 50mv voltage is lower than that before medication It is an effective drug for the treatment of arrhythmia.

[0007] The specific process is as follows:

[0008] a. Isolation of single cardiomyocytes: Isolation of single cardiomyocytes is obtained by the following methods:

[0009] After the experimental animal was anesthetized with 2% pentobarbital sodium, the chest was opened, the heart was quickly taken out, the aort...

specific Embodiment approach 3

[0021] Specific embodiment three: the screening process of this embodiment is as follows: the guinea pig is anesthetized with 2% pentobarbital sodium, and the chest is opened, and the heart is quickly taken out, and the aortic retrograde cannula is connected to the Langendorff perfusion device, and normal Tyrode's solution is used. About 8ml.min -1Continuous perfusion was performed for 5 minutes to wash away residual blood from the heart, and then perfused with calcium-free Tyrode's solution for about 10 minutes. After the heart stopped beating, the cardiac single cells were separated by perfusion with calcium-free solution containing 0.05% type II collagenase. When the heart became soft and the color became lighter, myocardial tissue was collected every 2 minutes. All the above operations were performed at a constant temperature of 37°C and filled with 95% O 2 +5%CO 2 The atrial muscle tissues removed at different times were placed in test tubes filled with KB solution, and...

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Abstract

The invention relates to a new use of cardiocyte M3 receptor and IKM3 for screening drugs to treat cardiac arrhythmias. The method of screening drugs using cardiocyte M3 receptor and IKM3 as the target has advantages of high accuracy, strong specificity, and simple operation. The drugs screened by means of cardiocyte M3 receptor and IKM3 has high specificity, and can obviously decrease the adverse reactions, promote the curative effect.

Description

Technical field: [0001] The present invention relates to a new application of cardiomyocytes, in particular to cardiomyocyte muscarinic acetylcholine receptor M3 subtype and IK M3 Use of currents as targets for screening antiarrhythmic drugs. Background technique: [0002] Arrhythmia seriously endangers human health and is one of the main causes of human death. Epidemiological studies have shown that 50% of heart failure patients die of arrhythmia. Although some antiarrhythmic drugs have improved the patient's condition, there are still obvious shortcomings: ① The total effective rate of antiarrhythmic drugs is not high, only 30% to 60%; ② Ineffective for some stubborn arrhythmias; ③ Long-term The application has a poor effect on improving the survival rate of patients; ④ prone to problems such as arrhythmia-induced side effects. Moreover, before there is no practical application, it is difficult for people to judge which drug has the effect of treating arrhythmia. When sc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
Inventor 杨宝峰王慧珍董德利
Owner HARBIN MEDICAL UNIVERSITY
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