Diacylhydrazine ligands for modulating the expression of exogenous genes in mammalian systems via an ecdysone receptor complex

摘要

The present invention relates to non-steroidal ligands for use in nuclear receptor-based inducible gene expression system, and a method to modulate exogenous gene expression in which an ecdysone receptor complex comprising: a DNA binding domain; a ligand binding domain; a transactivation domain; and a ligand is contacted with a DNA construct comprising: the exogenous gene and a response element; wherein the exogenous gene is under the control of the response element and binding of the DNA binding domain to the response element in the presence of the ligand results in activation or suppression of the gene.

主权项

1. A method of modulating the expression of a target gene in a host cell, wherein the host cell includes a first gene expression cassette comprising a first polynucleotide encoding a first polypeptide comprising:
(i) a transactivation domain; (ii) a DNA-binding domain; and (iii) a Group H nuclear receptor ligand binding domain; a second gene expression cassette comprising: (i) a response element capable of binding to said DNA binding domain; (ii) a promoter that is activated by the transactivation domain; and (iii) said target gene;the method comprising contacting said host cell with a compound of the formula: wherein X and X′ are independently O or S; Y is: (a) substituted or unsubstituted phenyl wherein the substituents are independently 1-5H, (C1-C4)alkyl, (C1-C4)alkoxy, (C2-C4)alkenyl, halo (F, Cl, Br, I), (C1-C4)haloalkyl, hydroxy, amino, cyano, or nitro; or (b) substituted or unsubstituted 2-pyridyl, 3-pyridyl, or 4-pyridyl, wherein the substituents are independently 1-4H, (C1-C4)alkyl, (C1-C4)alkoxy, (C2-C4)alkenyl, halo (F, Cl, Br, I), (C1-C4)haloalkyl, hydroxy, amino, cyano, or nitro; R1 and R2 are independently: H; cyano; cyano-substituted or unsubstituted (C1-C7) branched or straight-chain alkyl; cyano-substituted or unsubstituted (C2-C7) branched or straight-chain alkenyl; cyano-substituted or unsubstituted (C3-C7) branched or straight-chain alkenylalkyl; or together the valences of R1 and R2 form a (C1-C7) cyano-substituted or unsubstituted alkylidene group (RaRbC═) wherein the sum of non-substituent carbons in Ra and Rb is 0-6; R3 is H, methyl, ethyl, n-propyl, isopropyl, or cyano; R4, R7, and R8 are independently: H, (C1-C4)alkyl, (C1-C4)alkoxy, (C2-C4)alkenyl, halo (F, Cl, Br, I), (C1-C4)haloalkyl, hydroxy, amino, cyano, or nitro; and R5 and R6 are independently: H, (C1-C4)alkyl, (C2-C4)alkenyl, (C3-C4) alkenylalkyl, halo (F, Cl, Br, I), (C1-C4)haloalkyl, (C1-C4)alkoxy, hydroxy, amino, cyano, or nitro; wherein the number of carbon atoms, excluding those of cyano substitution, for either or both of groups R1 or R2 is greater than 4, and the number of carbon atoms, excluding those of cyano substitution, for the sum of groups R1, R2, and R3 is 10, 11, or 12.