Screening drugs disrupting macrophage migration inhibitory factor-cJun activation domain binding protein1 interaction involves comparing MIF-Jab1 interaction, in presence and absence of drug in MIF and Jab1 system

摘要

Screening (M1) candidate drugs which disrupt an interaction between MIF (macrophage migration inhibitory factor) and Jab1 (c-Jun activation domain binding protein) and/or interfere with MIF-Jab1-dependent action, involves detecting and then comparing interaction between MIF and Jab1 and/or a MIF-Jab1-dependent action, in MIF and Jab1 system, in the presence and absence of the candidate drug. Independent claims are also included for the following: (1) detecting (M2) an interaction between MIF and Jab1 involves: (a) providing a cell containing a reporter gene which expresses a reporter protein when the reporter gene is activated by an amino acid sequence, including a transcriptional activation domain when the transcriptional activation domain is in sufficient proximity to the reporter gene; (b) providing a first chimeric gene that is capable of being expressed in host cells, comprising a DNA sequence that encodes a first hybrid protein, the first hybrid protein comprising a DNA-binding that recognizes a binding site of the reporter gene in the cell and the MIF protein or its part; (c) providing a second chimeric gene that is capable of being expressed in the host cell, comprising a DNA sequence that encodes a second hybrid protein, the second hybrid protein comprising the transcriptional activation domain and the Jab1 protein or its part, where interaction between the MIF protein and the Jab1 in the cell causes the transcriptional activation domain to activate transcription of the reporter gene; (d) introducing the first chimeric gene and the second chimeric gene into the cell; (e) subjecting the cell to conditions under which the first hybrid protein and the second hybrid protein are expressed in sufficient quantity for the reporter gene to be activated; and (f) determining whether the reporter gene has been expressed to a degree greater than expression in the absence of the interaction between the MIF protein and the Jab1 protein; (2) preparing (M3) MIF involves contacting a first source containing MIF with a second source containing Jab1 under conditions allowing for the binding of MIF and Jab1 and then separating MIF from Jab1; (3) preparing (M4) Jab1 involves contacting a first source containing Jab1 protein with a second source containing MIF under conditions allowing for the binding of MIF and Jab1 and then separating Jab1 from MIF; (4) a protein complex (I) comprising all or a part of MIF in association with all or part of Jab1, optionally in association with p27Kip1 and/or c-Jun and/or c-Jun-amino-terminal kinase and/or steroid receptor coactivator 1 and/or integrin LFA-1 and/or progesterone receptor and/or glucocorticoid receptor; (5) a fusion protein (II) comprising all or part of MIF in conjunction with all or part of Jab1, optionally in conjunction with p27Kip1 and/or c-Jun and/or c-Jun-amino-terminal kinase and/or steroid receptor coactivator 1 and/or integrin LFA-1 and/or progesterone receptor and/or glucocorticoid receptor; (6) a nucleic acid sequence (III) encoding (I) or (II), or its complementary strand; (7) a vector (IV) comprising (III); (8) a host cell (V) comprising (IV); (9) an antibody (VI) or its fragment recognizing (I) and/or (II); (10) a kit comprising (I)-(V), the DNA sequence encoding Jab1, the DNA sequence encoding MIF and/or (VI); (11) a pharmaceutical or diagnostic composition comprising (I)-(V), the DNA sequence encoding Jab1, the DNA sequence encoding MIF and/or (VI), optionally in conjunction with a carrier; (12) use of the Jab1/Jab1 coding nucleotide sequence, to modulate the activity of MIF for treating MIF-related diseases, for preparing a drug for the diagnosis and/or treatment of MIF-related diseases, and for the diagnosis and/or treatment of MIF-related diseases; (13) a drug comprising Jab1 or its part, or a Jab1 coding nucleotide sequence or an antibody specifically recognizing Jab1, optionally in conjunction with a carrier; (14) use of MIF/MIF coding nucleotide sequence to modulate the activity of Jab1, for preparing a drug for the diagnosis and/or treatment of Jab1-related diseases, and for the diagnosis and/or treatment of Jab-related diseases; (15) a drug comprising MIF or its part or a MIF coding nucleotide sequence or an antibody specifically recognizing MIF, optionally in conjunction with a carrier; (16) a non-human mammalian cell (VII) exhibiting a modified MIF and Jab1 expression; (17) a non-human mammal containing (VII); (18) a human cell (VIII) exhibiting a modified MIF and Jab1 expression; and (19) a non-human mammal containing (VIII). ACTIVITY : Antibacterial; antiarthritic; antiinflammatory; vulnerary; cytostatic; auditory; dermatalogical; antidiabetic; anorectic; antiparkinsonian; immunosuppressive. No biodata is provided. MECHANISM OF ACTION : MIF/Jab1 interaction modulator; MIF or Jab1 activity modulator; gene therapy.