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Application of miRNA (micro-ribonucleic acid) miR-21 to glaucoma treatment

A technology of mirna-21 and .mirna-21 is applied in the application field of small molecule nucleic acid miR-21 in the treatment of glaucoma, which can solve the problem of common adverse reactions in the eyes of patients.

Active Publication Date: 2018-11-02
EYE & ENT HOSPITAL SHANGHAI MEDICAL SCHOOL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, judging from the preliminary clinical application effect, the effect of Ripasudi l on lowering intraocular pressure is only 15.5%, and the adverse reactions to the eyes of patients are more common

Method used

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  • Application of miRNA (micro-ribonucleic acid) miR-21 to glaucoma treatment
  • Application of miRNA (micro-ribonucleic acid) miR-21 to glaucoma treatment
  • Application of miRNA (micro-ribonucleic acid) miR-21 to glaucoma treatment

Examples

Experimental program
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Effect test

Embodiment 1

[0118] Effect of miR-21 on intraocular pressure

[0119] Combining miR-21mimics with transfection reagents in vivo, the drug was injected into the anterior chamber of normal mice by anterior chamber injection, and a negative control (Negative Control, NC) was given to the contralateral eye (the negative control was a nonsense random sequence ). Administration of mouse anterior chamber under microscope figure 1 shown.

[0120] Changes in intraocular pressure were measured with a tonometer before and after administration. When it was confirmed that the change in intraocular pressure was the most obvious, the mouse eyeball was completely removed, and the outflow rate of aqueous humor was measured using a mouse isolated eyeball perfusion model. The specific operation method is as follows:

[0121] Use a 33G needle to puncture the anterior chamber, and maintain constant pressure (12mmHg) perfusion for 30 minutes, and then measure the changes in the outflow and outflow rate of n...

Embodiment 2

[0125] Effect of miR-21 on cell permeability

[0126] Using porcine angular aqueous plexus (AAP) cells to simulate the Schlemm duct cells in the human trabecular meshwork pathway, using virus transfection to up-regulate the amount of intracellular miR-21, and measuring the transendothelial electric resistance (TEER) of the cells , according to the inverse ratio of transmembrane resistance to cell permeability, reflecting the resistance in the outflow pathway of aqueous humor.

[0127] The result is as Figure 5 Shown: Permeability increased after cells were transfected with viruses overexpressing miR-21mimics. 5A and 5B overexpressed miR-21mimics by virus transfection, and the transfection efficiency was 70%. Compared with transfection of empty virus (Negative Control group), miR-21mimics transmembrane cell resistance of 5C was significantly reduced. That is, up-regulation of miR-21mimics can increase the permeability of porcine Schlemm duct endothelial cells (p<0.01), and r...

Embodiment 3

[0129] miR-21 targets the trabecular pathway to achieve multi-target blood pressure reduction

[0130] Obtaining chamber angle tissue: The mice transfected for 24 hours were killed by cervical dislocation, and the eyeballs were taken out immediately. Use ophthalmic micro-scissors to cut along the equator of the eyeball, take out the crystal, gently pull out the iris with micro-tweezers, take out the root of the iris as much as possible, carefully cut off the cornea along the limbus, cut off the posterior structure about 0.5mm behind the limbus, and take the cornea The structure between the edge and the back is the angle tissue. Histologically, it contains trabecular meshwork, Schlemm's canal, and to a lesser extent the cornea, iris root, and external sclera.

[0131] Protein pathway verification: fresh chamber angle tissues transfected with miR-21mimics and NC were taken, lysed with RIPA lysate on ice for 1 hour, and ultrasonically shattered. After the protein concentration ...

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Abstract

The invention provides an application of miRNA (micro-ribonucleic acid) miR-21 to glaucoma treatment and specifically provides an application of an active component. The active component is selected from the following groups: (a) miRNA from miRNA-21; (b) precursor miRNA; (c) polynucleotide; (d) an expression vector; (e) an agonist of miRNA in (a); the active component is used for preparing pharmaceutical composition which is used for preventing and / or treating glaucoma and / or reducing IOP (intraocular pressure). About 20% of pressure reduction can be realized by miRNA miR-21, and the pressurereduction effect keeps for 48 h which is longer than that of most drugs clinically.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of small molecule nucleic acid miR-21 in treating glaucoma. Background technique [0002] Glaucoma is the world's first irreversible blinding eye disease. The disease is a type of irreversible blinding eye disease with characteristic optic nerve damage and visual field changes. Elevated intraocular pressure is the main risk factor for its occurrence and disease progression. The exact pathogenesis of elevated IOP in patients with the highest incidence of primary open angle glaucoma (POAG) remains unclear. The structure of the aqueous humor outflow channel in these patients is normal, and its pathogenesis may be due to the failure of the intraocular pressure (IOP) self-regulation mechanism, which leads to the increase of intraocular pressure and eventually damages the optic nerve. IOP-lowering therapy is the only proven effective treatment for glaucoma. Cu...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61P27/06A61P27/02
CPCA61P27/02A61P27/06A61K31/7105
Inventor 雷苑陈君毅孙兴怀檀宸
Owner EYE & ENT HOSPITAL SHANGHAI MEDICAL SCHOOL FUDAN UNIV
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