Metabolic labeling and molecular enhancement of biological materials using bioorthogonal reactions

A bioorthogonal, bioactive molecular technology for metabolic labeling and molecular enhancement of biomaterials using bioorthogonal reactions to address issues affecting short- and long-term graft outcomes

Active Publication Date: 2019-05-03
THE GENERAL HOSPITAL CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, ischemia / reperfusion injury is a common consequence after organ transplantation and affects short-term as well as long-term graft outcome

Method used

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  • Metabolic labeling and molecular enhancement of biological materials using bioorthogonal reactions
  • Metabolic labeling and molecular enhancement of biological materials using bioorthogonal reactions
  • Metabolic labeling and molecular enhancement of biological materials using bioorthogonal reactions

Examples

Experimental program
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preparation example Construction

[0357] The preparation of the compounds provided herein may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and selection of appropriate protecting groups can be readily determined by those skilled in the art. The chemical properties of protecting groups can be found, for example, in P.G.M. Wuts and T.W. Greene, Protective Groups in Organic Synthesis [Protective Groups in Organic Synthesis], 4th Edition, Wiley & Sons, Inc. [Wiley & Sons], New York (2006) turn up.

[0358] In some embodiments, growth factors such as basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) can be obtained by using alkyne-containing or alkyne-containing N-hydroxysuccinimide esters (NHS esters). Cycloalkyne reagents are conjugated to alkyne or cycloalkyne (eg, DBCO) functional groups. A PEG linker or an alkylene linker, or both, of various lengths can be introduced between the alkyne or cycloalkyne functional...

example

[0503] Materials and General Methods

[0504] In vivo metabolic engineering and organ / tissue decellularization

[0505] All animal experiments were approved by the Massachusetts General Hospital Institutional Animal Care and Use Committee and performed in accordance with the Animal Welfare Act. Male Sprague-Dawley rats (100-125 g, Charles River Laboratories) were given metabolic labeling reagents (Ac4GalNAz, Ac4GlcNAz or Ac4ManNAz) (30 mg / day, Click Chemistry Tools) daily via intraperitoneal injection for three days. One day after the last administration of metabolic labeling reagents, organs were harvested from animals and decellularized by perfusion using the following conditions: 0.1% SDS through the pulmonary artery (PA) for the lungs; 1% SDS retrograde coronary perfusion through the ascending aorta for the heart 1% SDS is delivered to the kidneys via the renal artery; and 1% SDS is delivered to the liver via the inferior vena cava (where the superior vena cava is ligated...

example 1- 3

[0520] Example 1 - Comparison of metabolic labeling efficiencies of three azide-labeled sugars (Ac4GalNAz, Ac4GlcNAz and Ac4ManNAz)

[0521] The first step in the presently described method and procedure is to generate ligands (azide tags) on decellularized organ / tissue scaffolds by metabolic labeling using azide-tagged sugars for chemoselective ligation (click reaction). ). In the method described, azide-labeled galactosamine (Ac4GalNAz) is used to metabolically label decellularized native organ / tissue scaffolds. Using decellularized lung scaffolds as a model, it was demonstrated that Ac4GalNAz exhibited higher labeling efficiency compared to other commercially available azide-labeled sugars such as Ac4GlcNAz and Ac4ManNAz.

[0522] exist figure 2 In A, images showing staining for azide labeling (purple) and the ECM component laminin (green) on decellularized rat lungs after 3 days of metabolic labeling in donor rats. Using biotin-alkyne (via click reaction) (Sigma-Aldri...

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Abstract

The present application provides methods of functionalizing an organ or tissue of a mammal by administering a nutrient (e.g., peracetylated N-azido galactosamine Ac4GalNAz) to the mammal or by culturing an organ or tissue in a bioreactor containing such nutrient. The present application also provides methods of selectively functionalizing extracellular matrix (ECM) of an organ or tissue of a mammal by administering a nutrient (e.g., peracetylated N-azido galactosamine Ac4GalNAz) to the mammal. In some aspects, the present application provides a decellularized scaffold of a mammalian organ or tissue comprising an extracellular matrix, wherein the extracellular matrix of the decellularized scaffold is functionalized with a chemical group that is reactive in a bioorthogonal chemical reaction,such as an azide chemical group. The present application also provides biological prosthetic mesh and mammalian organs and tissues for transplantation prepared according to the methods of the application.

Description

[0001] priority statement [0002] This application claims the benefit of U.S. Provisional Application No. 62 / 350,259, filed June 15, 2016. The entire contents of the above documents are hereby incorporated by reference. technical field [0003] The present invention relates to metabolic labeling and further functionalization of mammalian organs and tissues. Background technique [0004] Prosthetic mesh is a common solution to many clinical problems, the most common of which is hernia repair. While synthetic prosthetic meshes are strong and offer low recurrence rates, an increase in mesh-related complications, including infection, has been demonstrated. (Darehzereshki, A. et al. World J. Surg. 38, 40-50 (2014)) Synthetic meshes are contraindicated in cases of active Relatively contraindicated in patients or in procedures where there is a high risk of contamination. [0005] Bioprosthetics have been developed using a variety of tissue sources and processing techniques. M...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/14A61K31/715C12N5/07C12N5/02C12N5/00A61N1/30C07D233/88
CPCA61L27/56A61L27/54A61L27/3826A61L27/3813A61L27/383A61L27/3808A61L27/3633A61L27/3683A61K31/655A61K31/7008A61K31/7004A61P43/00C12N5/0602A61L2300/256A61L2300/406A61L2300/414A61L2300/42A61K35/22A61K35/34A61K35/36A61K35/407A61K35/42A61K35/44C07H13/04A61K9/0019C12N5/06
Inventor H·C·奥特任息J·P·布鲁姆T·K·拉加伯
Owner THE GENERAL HOSPITAL CORP
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