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Use of lysine-specific demethylase 1 (LSD1) inhibitor

A demethylase, specific technology, applied in the field of hearing protection for sensorineural deafness caused by aminoglycosides, and the hearing protection effect of LSD1 inhibitors for sensorineural deafness

Active Publication Date: 2014-09-17
EYE & ENT HOSPITAL SHANGHAI MEDICAL SCHOOL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no relevant report on the protective effect of lysine-specific demethylase 1 (LSD1) inhibitors on the hearing of sensorineural deafness

Method used

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  • Use of lysine-specific demethylase 1 (LSD1) inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Establishment of a zebrafish lateral line system hair cell injury model

[0074] The Brn3c:mGFP transgenic zebrafish (Eaton-Peabody Laboratory, Harvard University Eye and Ear Hospital, USA) was used in this study. Zebrafish were grown in a constant temperature environment of 28.5°C, with a photoperiod of 14 hours of light / 10 hours of darkness, and the breeding program was carried out according to the "Zebrafish Book" (http: / / www.zfin.org). The specific operation is as follows:

[0075] ① Expose 5 days post fertilization (5dpf) zebrafish to zebrafish feeding water containing 400 μM neomycin for 1 hour;

[0076] ② Rinse 3 times in fresh zebrafish rearing water;

[0077] ③ 0.02% MS-222 anesthetized for 5 minutes;

[0078] ④ Fix with 4% paraformaldehyde at room temperature for 2 hours;

[0079] ⑤ Rinse with 0.01M PBS for 5min×3;

[0080] ⑥1:800DAPI at room temperature for 20min;

[0081] ⑦Seal the slide and observe the hair cell damage in the lateral line sy...

Embodiment 2

[0082] Embodiment 2 utilizes zebrafish model to carry out drug screening

[0083] 1) S2101-neomycin treatment: zebrafish embryos developed to 5dpf before drug treatment: including control group (DMSO20μM) and S2101 experimental group (5μM, 10μM, 20μM). After drug treatment for 2 hours, neomycin was added to injure for 1 hour (400 μM), rinsed 3 times in fresh zebrafish feeding water, anesthetized with MS-222 for 5 minutes, fixed with 4% paraformaldehyde at room temperature for 2 hours; rinsed with 0.01M PBS for 5 minutes×3; 1:800 DAPI Room temperature for 20 minutes; cover the slides, observe and count under a fluorescence microscope, and observe the survival of lateral liner hair cells. Among them, n=50-70, statistical results were analyzed with SPSS11.5 statistical software ( figure 2 ).

[0084] 2) CBB1007-neomycin treatment: zebrafish embryos developed to 5dpf before drug treatment: including control group (DMSO100μM) and CBB1007 experimental group (20μM, 50μM, 100μM). ...

Embodiment 3

[0085] Example 3 Establishment of mouse cochlear hair cell injury model

[0086] 1) In vitro model

[0087] ① Isolation of auditory epithelium from neonatal mouse cochlea

[0088] ② In vitro tissue culture of neonatal mouse auditory epithelium

[0089] ③Drug treatment

[0090] The newborn mouse cochlear auditory epithelium was isolated and adhered to the wall for 2 hours. After the specimen adhered to the wall, it was added to serum-free DMEM / F12 medium containing S2101 or CBB1007 for co-culture for 24 hours, and then added to serum-free DMEM / F12 containing 1mM neomycin. The F12 medium was co-cultured for 4 hours. Rinse 3 times in serum-free DMEM / F12. Finally, serum-free medium was added for co-cultivation for 24 hours, and the hair cell injury model of the cochlear basement membrane showed a gradient change from the bottom circle to the top circle ( image 3 , the bottom circle is severe, the top circle is slight).

[0091] 2) In vivo model

[0092] The experiment adop...

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Abstract

The invention relates to a use of a lysine-specific demethylase 1 (LSD1) inhibitor. Zebra fish and mouse are used as model animals and used for building hair cell damage and protection research animal models. A research result proves that through utilization of the LSD1 inhibitor on the whole body and round window local part, a histone H3K4 dimethylation expression level is substantially improved and cochlear hair cell Cleaved Caspase-3 expression is substantially reduced so that hair cell damage is reduced and hearing protection is realized and thus it is shown that the LSD1 inhibitor can produce hearing protection in sensorineural deafness treatment. The use provides a novel approach for searching, preventing and treating sensorineural deafness and has a clinical application value.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to the new use of lysine-specific demethylase 1 (LSD1) inhibitors, in particular to the hearing protection effect of LSD1 inhibitors on sensorineural deafness, especially to amino The use of glycoside drugs in hearing protection for sensorineural deafness. Background technique [0002] Sensorineural deafness is a common and frequently-occurring disease in otolaryngology. In recent years, the incidence has been on the rise. It not only brings physical and psychological pain to patients and their families, but also has a serious impact on social and economic development. According to the second national sample survey of disabled people, there are 20.04 million hearing disabled people and 1.27 million speech disabled people in my country. This is a social group with a large number and prominent difficulties. Hearing impairment and deafness have become one of the important factors a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/496A61P27/16A61P25/02
Inventor 李华伟于慧前何英姿孙珊黎奥柴人杰
Owner EYE & ENT HOSPITAL SHANGHAI MEDICAL SCHOOL FUDAN UNIV
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