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A new drug based on sphk1 inhibitor combined with aav9-sphk2 virus and its application

An inhibitor, aav9-sphk2 technology, is applied in the direction of drug combination, drug delivery, and active ingredients of heterocyclic compounds, which can solve the problems of heart failure, heart failure, and low regeneration ability of cardiomyocytes, so as to reduce cardiac fibrosis and promote Cardiac regenerative repair, enhancement of therapeutic and preventive effects

Active Publication Date: 2022-04-22
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At the same time, due to the extremely low regenerative capacity of adult cardiomyocytes, after myocardial infarction, the injured myocardium successively undergoes inflammatory reactions, extracellular matrix deposition, collagen formation, and finally myocardial fibrosis, forming fibrotic scar tissue. The existence of cardiac scar tissue will Causes loss of heart pumping ability and lack of circulatory function, leading to heart failure and death
[0004] At present, the drugs and methods for the treatment of heart disease, especially ischemic heart disease, are still very limited. Drugs, coronary artery catheterization and revascularization have significantly improved cardiac function after myocardial infarction, but also lead to more and more More and more surviving patients have permanent structural damage to the heart, which may lead to serious consequences such as heart failure, so it is necessary to develop new drugs for the treatment of myocardial injury

Method used

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  • A new drug based on sphk1 inhibitor combined with aav9-sphk2 virus and its application
  • A new drug based on sphk1 inhibitor combined with aav9-sphk2 virus and its application
  • A new drug based on sphk1 inhibitor combined with aav9-sphk2 virus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] In this example, an in vitro experiment was used to explore the activation effect of SphK1 on cardiac fibroblasts. The specific test method is as follows.

[0033] 1. Isolation of primary rat neonatal cardiac fibroblasts

[0034] After sterilizing the suckling rats (5 rats) with 75% alcohol, pinch the back skin of the suckling rats tightly with the left hand, cut the sternum with scissors from the left edge of the xiphoid process, expose the heart, take it out with tweezers and wash it in PBS. In another glass plate, remove the connective tissue and atrium as much as possible; cut the heart into pieces with scissors, add 5mL of digestion solution, transfer it to a graduated bottle with a straw, place it in a 37°C water bath, and adjust to an appropriate speed for digestion. Digest for 30 minutes for the first time, collect the digested supernatant, transfer it to a 20mL centrifuge tube containing DMEM containing 10% FBS, then add 5mL of digestion solution and gently blo...

Embodiment 2

[0047] In this example, in vitro experiments are used to explore the effect of SphK1 inhibitors on inhibiting the activation of cardiac fibroblasts, and the specific experiment process is as follows.

[0048] 1. Isolation of primary rat neonatal cardiac fibroblasts

[0049] Cardiac fibroblasts were isolated by referring to the method for isolating cardiac fibroblasts of primary neonatal rats in Example 1.

[0050] 2. Test method

[0051] a. Control group: the primary cultured neonatal mouse cardiac fibroblasts were washed twice with PBS, added to DMEM basal culture medium and cultured to 80% cell confluence, and then cultured with serum-free basal culture medium for 24 hours, replaced The DMEM basal culture solution containing 10% FBS was used as the control group (Ctrl), which was cultured synchronously with the experimental group.

[0052] b. Experimental group: the primary cultured neonatal rat heart fibroblasts were inoculated on a 96-well culture plate at a certain dens...

Embodiment 3

[0060] In this example, an in vitro experiment was conducted to investigate the effect of the Ad-Sphk2 virus expressing sphingosine kinase 2 on the proliferation of neonatal rat cardiomyocytes. The specific experiment process is as follows.

[0061] 1. Isolation of primary rat neonatal rat cardiomyocytes

[0062] After sterilizing the suckling rats (5 rats) with 75% alcohol, pinch the skin on the back of the suckling rats with the left hand, cut the sternum with scissors from the left edge of the xiphoid process, expose the heart, take it out with tweezers and wash it in PBS. In another glass plate, remove the connective tissue and atrium as much as possible; cut the heart with scissors, add 5mL of digestion solution, transfer it to a graduated bottle with a straw, place it in a 37°C water bath, and adjust to an appropriate speed for digestion. Digest for 30 minutes for the first time, collect the digested supernatant, transfer it to a centrifuge tube containing 20 mL of DMEM ...

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Abstract

The invention belongs to the field of biomedicine technology, relates to the field of heart disease drug development, and more specifically relates to a new drug based on SphK1 inhibitor combined with AAV9-SphK2 virus and its application. The new drug includes SphK1 inhibitor and AAV9-SphK2 virus. SphK1 inhibitor alleviates cardiac fibrosis, promotes endogenous cardiomyocyte proliferation through AAV9-SphK2 virus, combined with drug-gene therapy, can significantly improve cardiac function and fibrosis after myocardial infarction in adult mice, greatly It can be used as a new drug for the prevention and treatment of heart disease such as ischemic heart disease by promoting the regeneration and repair ability of the heart after myocardial infarction to a certain extent.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, relates to the field of drug development for heart disease, and more specifically relates to a new drug based on SphK1 inhibitor combined with AAV9-SphK2 virus and its application. Background technique [0002] Cardiovascular disease is a public health problem with the highest mortality rate in the world. Heart failure caused by ischemic heart disease is one of the important factors that endanger human health, and cardiomyocyte loss is the root cause of heart failure. Studies have found that adult mammalian cardiomyocytes have a certain proliferation ability, but it is far from enough to supplement the loss of cardiomyocytes caused by myocardial infarction and other myocardial injuries. In recent years, many studies have also confirmed the possibility of endogenous myocardial regeneration, and promoting the proliferation of cardiomyocytes is an important means to activate endogenous myocardia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/06A61K31/40A61K48/00A61P9/00A61P9/10A61P9/04
CPCA61K45/06A61K31/40A61K48/0008A61K48/005A61K9/0019A61P9/00A61P9/10A61P9/04A61K2300/00
Inventor 曹楠纪晓倩
Owner SUN YAT SEN UNIV
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