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A kind of pharmaceutical composition targeting circRNA and its application

A composition and drug technology, applied in the field of biomedicine, to achieve the effects of environmental friendliness, simple preparation process, and obvious drug effect

Active Publication Date: 2018-06-22
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No research has revealed that circRNA plays a role in the process of cardiomyocyte apoptosis. Finding the circRNA specifically expressed in the heart and participating in cardiomyocyte apoptosis, and clarifying its mechanism of action, is helpful for the development of siRNA targeting circRNA in the diagnosis and treatment of heart diseases. The application in this aspect has great significance and broad prospects

Method used

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  • A kind of pharmaceutical composition targeting circRNA and its application
  • A kind of pharmaceutical composition targeting circRNA and its application
  • A kind of pharmaceutical composition targeting circRNA and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: circRNA expression during myocardial ischemia and cardiomyocyte apoptosis induced by hypoxia

[0021] The present embodiment adopts the experimental model of cardiomyocyte apoptosis, and cultures the primary cardiomyocytes of rat suckling rats by conventional methods, cultivates them for different times in a hypoxic incubator (oxygen concentration is lower than 1%), extracts RNA, and detects with real-time fluorescent quantitative PCR technology. The expression level of circRNA MFACR, such as figure 1A shows that circRNA MFACR is significantly up-regulated when cardiomyocytes undergo apoptosis within 4h-8h of hypoxia treatment, where Control is the A / R control group;

[0022] In this example, rat coronary artery ligation was used to establish a myocardial ischemia model. Myocardial tissues in ischemic and non-ischemic areas were collected at different times of ischemia, total RNA was extracted, and the expression level of circRNA MFACR was detected by real-ti...

Embodiment 2

[0023] Example 2: MFACR-siRNA inhibits cardiomyocyte apoptosis induced by hypoxia

[0024] In this example, primary cultured cardiomyocytes were used as a model, and the cultured primary cardiomyocytes were transfected with the MFACR-siRNA adenovirus vector. After 24 hours of transfection, the treated cells were cultured in a hypoxic incubator for 3 hours. Myocardial hypoxia induced cell apoptosis, and apoptosis detection kit (TUNEL method) was used to detect myocardial cell apoptosis. Comparing the four groups of cardiomyocytes with different treatments (such as figure 2 As shown in A or 2B, from left to right is the cardiomyocyte group without hypoxia treatment; the hypoxia treatment cardiomyocyte group, the transfection of MFACR-sc adenovirus vector and the hypoxia treatment cardiomyocyte group, namely the negative control group, the transfection Changes of mitochondrial fission and cell apoptosis in hypoxia-treated cardiomyocytes after transfection with MFACR-siRNA adeno...

Embodiment 3

[0025] Example 3: MFACR-siRNA inhibits cardiomyocyte apoptosis in myocardial ischemia injury

[0026] In this example, C57 / BL6 mice were used as the experimental objects, with 8 mice in each group. The measured results were statistically analyzed with one-way ANOVO statistical software, and 30 mg / kg MFACR-siRNA and its Negative control MFACR-sc, mice were weighed and anesthetized, fixed in supine position, sheared and the operation area was disinfected with iodine. Make a midline incision of the trachea in the neck and intubate, connect the animal ventilator for positive pressure ventilation, cut the skin longitudinally about 3 cm at the left edge of the sternum and the heartbeat, bluntly separate the subcutaneous tissue and muscles layer by layer, open the chest, and be careful Lift the pericardium and cut it open to fully expose the heart; dilute MFACR-siRNA and MFACR-sc in phosphate buffered saline respectively, with a final volume of 200 μL, enter the artery root from the ...

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Abstract

The invention discloses a pharmaceutical composition of targeting circRNA (circular Ribonucleic Acid). The pharmaceutical composition comprises MFACR-siRNA (small interfering Ribonucleic Acid) nucleotide, a pharmaceutically acceptable carrier or virus carrier and an auxiliary material; the carrier is one or more of chitosan, cholesterol, nanoparticles and liposome, preferably, the liposome; the auxiliary material is one or more of mannitol, a phosphate buffering solution and saline, preferably, the phosphate buffering solution; the content of the MFACR-siRNA is 0.8 to 1.5 micrograms; the mass ratio of the MFACR-siRNA to the liposome is 1 to 1.25; the mass ratio of the carrier to the auxiliary material is 1 to 200. The pharmaceutical composition is used for treating and preventing myocardial infarction, myocardial ischemial injury, myocardial hypertrophy and myocardial fibrosis, and is dosed in an oral administration or injection manner; the injection is intravenous injection, muscle injection, intracoronary injection or direct myocardial injection. Compared with the prior art, the selected raw materials are scientific and reasonable; a preparation process is simple and the drug action is obvious; the application range is wide, the utilization is safe and reliable and the application is environment-friendly.

Description

Technical field: [0001] The invention belongs to the technical field of biomedicine, and relates to a new nucleic acid drug targeting endogenous circRNA and its use, especially a pharmaceutical composition containing MFACR-siRNA nucleotides targeting circRNA MFACR and its role in the prevention or treatment of heart disease use in . Background technique: [0002] Cardiovascular disease is the number one killer of human health. More than 17 million people die of cardiovascular disease in the world every year, and its mortality rate is close to the sum of all cancer mortality rates. It is a major threat to human health and life. With the improvement of human living standards and the change of diet structure, the mortality rate of cardiovascular disease is on the rise, surpassing cancer as the number one cause of death. According to the latest statistical results released recently, the prevalence of hypertension among residents aged 18 and above is 18.8%, and it is estimated t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7088A61K47/36A61K47/28A61K47/10A61K47/02A61K47/46A61P9/10A61P9/00A61K9/19
CPCA61K9/0056A61K9/19A61K31/7088A61K47/02A61K47/10A61K47/28A61K47/36A61K47/46
Inventor 王昆李培峰周露玙刘翠云
Owner QINGDAO UNIV
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