Imaging of enzymatic activity

Details Imaging of enzymatic activity Imaging of enzymatic activity

C - Chemistry, Metallurgy

12

Q

C12Q 1/34 (2006.01) C12Q 1/26 (2006.01) C12Q 1/37 (2006.01) C12Q 1/48 (2006.01) G01R 33/465 (2006.01)

Patent

CA 2425873

The invention features methods of detecting enzymatic activity (e.g., in a magnetic resonance image). The methods include: (I) providing a monomeric substrate (e.g., a substrate that is polymerizable in the presence of an enzyme or as a result of an enzyme-catalyzed reaction), having the generic structure X-Y-Z, where X includes a chelator moiety having a chelated paramagnetic or supermagnetic metal atom or iron, Y includes a linker moiety (e.g., to provide a covalent or non-covalent chemical bond or bonds between X and Z), and Z includes a polymerizing moiety; (2) contacting the substrate with a target tissue, wherein the substrate undergoes polymerization to form a paramagnetic or supermagnetic polymer, the polymerization being catalyzed by an enzyme in an extracellular matrix or bound to the surfaces of cells of the target tissue; and (3) detecting an increase in relativity for the polymer relative to an equivalent amount of unpolymerized substrate. The invention also features substrate compositions. Figure (1) is a synthetic pathway used for synthesis of an enzyme-responsive, paramagnetic monomeric substrate.

L'invention concerne des méthodes de détection de l'activité enzymatique (par ex. l'imagerie par résonance magnétique). De façon générale, ces méthodes consistent à: (I) fournir un substrat monomère (par ex. un substrat polymérisable en présence d'une enzyme ou suite à une réaction catalysée par une enzyme), présentant la structure générique X-Y-Z, dans laquelle X comprend une fraction de chélateur contenant un ion ou un atome métallique paramagnétique ou superparamagnétique chélaté, Y comprend une fraction de liant (par ex. pour assurer une liaison (ou des liaisons) chimique covalente ou non covalente entre X et Z), et Z comprend une fraction de polymérisation; (2) mettre en contact le substrat avec un tissu cible, le substrat subissant une polymérisation pour former un polymère paramagnétique ou superparamagnétique, la polymérisation étant catalysée par une enzyme dans une matrice extracellulaire ou liée aux surfaces des cellules du tissu cible; et enfin, (3) détecter l'augmentation de relaxivité du polymère correspondant à une quantité équivalente de substrat non polymérisé. Par ailleurs, l'invention concerne des compositions de substrat.

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