Method for detecting of a compound that mimics, potentiates...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2335557

A method for detection of a compound that mimics, potentiates or inhibits the physiological effect of leptin, which method comprises: (a) for a compound which mimics the physiological effect of the leptin, assessing the effect of the compound upon a leptin activated signal transducer and activator of transcription (STAT) DNA response element coupled to a reporter gene; or (b) for a compound which potentiates or inhibits the physiological effect of leptin, assessing the effect of the compound upon the response provided by leptin upon a leptin activated STAT DNA response element coupled to a reporter gene; wherein, the response element and the reporter are expressed in the human Caucasian colon epithelial cell line (CACO3-2) or the response element and the reporter are expressed in CACO-2 cells, which cell line (the engineered cell line) is also transfected with a polypeptide which is capable of mediating or enhancing the stimulation by leptin of a leptin activated STAT DNA response element and contains the appropriate STAT proteins.

L'invention concerne un procédé de détection d'un composé simulant, renforçant ou inhibant l'effet physiologique de la leptine, comprenant les étapes ci-après : (a) pour un composé simulant l'effet physiologique de la leptine, évaluation de l'effet du composé sur un élément de réponse à transducteur de signal et activateur de transcription (STAT)ADN, accouplé à un gène rapporteur, activé par la leptine, ou (b) pour un composé renforçant ou inhibant l'effet physiologique de la leptine, évaluation de l'effet du composé sur la réponse fournie par la leptine sur un élément de réponse activé par la leptine STAT ADN, accouplé à un gène rapporteur, caractérisé en ce que l'élément de réponse et le rapporteur sont exprimés dans la lignée cellulaire épithéliale du côlon caucasien humain (CACO-2), ou en ce que l'élément de réponse et le rapporteur sont exprimés dans des cellules CACO-2, laquelle lignée cellulaire (lignée cellulaire technique) est également transférée avec un polypeptide à mediation ou augmentant la stimulation par la leptine d'un élément de réponse STAT ADN activé par la leptine et renfermant les protéines STAT appropriées.

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