Use of a composition comprising 3,6,7-trimethyllumazine for preventing, ameliorating or treating mmp-9 associated conditions and inflammation

A technology of trimethyldioxytetrahydropteridine and MMP-9, applied in containing 3, can solve problems such as anti-inflammatory effects that are not discussed

Pending Publication Date: 2022-02-25
COMVITA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It does not discuss specific anti-inflammatory effects

Method used

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  • Use of a composition comprising 3,6,7-trimethyllumazine for preventing, ameliorating or treating mmp-9 associated conditions and inflammation
  • Use of a composition comprising 3,6,7-trimethyllumazine for preventing, ameliorating or treating mmp-9 associated conditions and inflammation
  • Use of a composition comprising 3,6,7-trimethyllumazine for preventing, ameliorating or treating mmp-9 associated conditions and inflammation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0322] Example 1 - Fluorescence Measurement

[0323] In this example, a fluorescent inhibitor screen provides a rapid, sensitive and high-throughput method to identify potential inhibitors of MMP-9.

[0324] Methods and materials

[0325] MMP-9 inhibitor screening assay (fluorometric assay) kit was purchased from Abcam (Melbourne, Australia). The fluorescent kit contains recombinant MMP-9 enzyme, MMP inhibitor NNGH (N-isobutyl-N-[4-methoxybenzenesulfonyl]glycylhydroxamic acid), MMP fluorescent substrate dissolved in DMSO reagents, fluorescence assay buffer, and 96-well transparent microplates.

[0326] Inhibitory activity against MMP-9 was assessed using a commercially available MMP-9 inhibitor screening assay kit. MMP-9 activity was expressed as a change in fluorescence intensity measured using a SpectraMax iD3 multimode microplate reader (Molecular Devices, San Jose, USA).

[0327] This assay uses a FRET-labeled (fluorescence resonance energy transfer) substrate that c...

Embodiment 2

[0334] Example 2 - Colorimetric Assay

[0335] In this example, the biological activity of 3,6,7-trimethyldioxotetrahydropterin was further studied using the MMP-9 Colorimetric Inhibitor Screening Kit.

[0336] Methods and materials

[0337] MMP-9 inhibitor screening assay (colorimetric) kit was purchased from Abcam (Melbourne, Australia). The kit contains recombinant MMP-9 enzyme, MMP inhibitor NNGH, MMP chromogenic substrate, colorimetric assay buffer and 96-well transparent microplate.

[0338] The colorimetric kit uses a thiopeptide as a chromogenic substrate (Ac-PLG-[2-Mercapto-4-methyl-pentanoyl]-LG-OC2H5), which can be hydrolyzed by MMPs to generate thiols. This intermediate is further reacted with DTNB [5,5'-dithiobis(2-nitrobenzoic acid), Ellman's reagent] to form 2-nitro-5-thiobenzoic acid, which can be detected by absorbance at 412 nm . Changes in absorbance were measured using a SpectraMax iD3 multimode microplate reader (Molecular Devices, San Jose, USA). A...

Embodiment 3

[0345] Example 3: Gelatin gel zymography

[0346] In order to confirm the inhibition of MMP-9 by 3,6,7-trimethyldioxotetrahydropteridine, the inventors performed gelatin gel zymography to detect the activity of MMP-9. Gelatin gel zymography is uniquely designed to detect active MMP-9 (gelatinase) due to its ability to digest gelatin.

[0347] Methods and materials

[0348] Novex TM A 10% zymogram plus (gelatin) protein gel (15 wells) was purchased from Thermo Fisher Scientific (Auckland, New Zealand). All chemicals required for zymography were also purchased from Thermo Fisher Scientific, these included Novex TMSharp pre-stained protein standard, Novex tris-glycine SDS sample buffer, Novex tris-glycine SDS electrophoresis buffer, Novex zymography refolding buffer and Novex zymography developing buffer. From Sartorius Purify double distilled water in the Pro (18.2MΩcm) purification system. Gelatin gel zymography was performed as an independent technique to demonstrate ...

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Abstract

The invention relates to compositions comprising 3,6,7-trimethyllumazine, methods and uses thereof in preventing, ameliorating or treating inflammation and / or preventing, ameliorating or treating conditions associated with inflammation. More particularly, though not solely, the invention relates to compositions comprising 3,6,7-trimethyllumazine and methods of use thereof in preventing, ameliorating or treating MMP-9 associated conditions, such as inflammation of the gastrointestinal tract and / or inflammatory conditions associated with the gastrointestinal tract.

Description

[0001] related application [0002] This application claims priority from New Zealand Patent Application No. 755138, which is hereby incorporated by reference. technical field [0003] The present invention relates to a composition comprising 3,6,7-trimethyldioxotetrahydropteridine, its method and use in preventing, ameliorating or treating inflammation and / or preventing, ameliorating or treating disorders related to inflammation. More specifically, but not exclusively, the present invention relates to compositions comprising 3,6,7-trimethyldioxotetrahydropteridine and methods of use thereof in preventing, improving or treating MMP-9-related disorders, such as Preventing, ameliorating or treating inflammation of the gastrointestinal tract and / or inflammatory conditions associated with the gastrointestinal tract. Background technique [0004] Inflammation associated with the immune system can be beneficial, but not always. Generally considered a negative reaction or a react...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/519A61K35/644A61K36/61A61P29/00A61P1/04
CPCA61K35/644A61P29/00A61P1/04A61K31/519A61K36/61
Inventor R·托塔M·布林布尔J·C·伊万斯J·M·C·斯蒂芬斯K·鲁迈斯B·林
Owner COMVITA
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