Specific targeting of RNA expanded repeat sequences
The present invention provides small molecule compounds that can form covalent adducts with specific sequences of RNA, such as the hairpin loop r(CUG) exp sequence which is a cause of myotonic dystrophy type 1 (DM1), or the r(CGG) exp sequence which is a cause of fragile X-associated tremor/ataxia syndrome (FXTAS); to methods of making the small molecule compounds; and to methods of using the small molecular compounds in the treatment of DM1 or of FXTAS in patients afflicted therewith. The invention further provides a method for identifying an RNA target of a small molecule drug in vivo, using a small molecule drug conjugated to an RNA-reactive crosslinker group and a reporter group, contacting a cell or nucleic acid extract with the small molecule drug conjugate, then separating RNA targets crosslinked to the small molecule drug conjugate by interaction of the affinity group with a complementary affinity group.
1. A method of identifying an RNA target of a small molecule drug in vivo, comprising contacting a living cell with a small molecule drug conjugate comprising an RNA targeting group, a crosslinking group reactive with the RNA, and an affinity group, then, optionally, activating the crosslinking group, then, separating RNA targets crosslinked to the small molecule drug conjugate by an interaction of the affinity group with a complementary affinity group;
wherein the small molecule drug conjugate is a compound of formula (I)
wherein
each individually selected R or R′ is H or (C1-C6)alkyl;
n1 is 2, 3, 4, 5, or 6;
Z is a (C1-C3)alkylene group, optionally substituted with a reporter or affinity group;
L is a linker group comprising an optionally substituted (C1-C6)alkylene, wherein one or two carbon atoms is optionally replaced by O;
XL is an RNA-reactive crosslinking group;
each individually selected Ht is an RNA-targeting group;
or any tautomer thereof; or any salt thereof, or any stereoisomer thereof.
2. The method of claim 1 wherein for the compound of formula (I), the crosslinking group is an alkylating group.
3. The method of claim 2 wherein for the compound of formula (I) the alkylating group is a chlorambucil derivative.
4. The method of claim 1 wherein for the compound of formula (I) the crosslinking group is a triggered crosslinker using photochemical or shape-triggered catalysis.
5. The method of claim 1 wherein for the compound of formula (I), Z is an alkylene group substituted with an affinity group.
6. The method of claim 5 wherein the affinity group is a biotin-comprising moiety.
7. The method of claim 1 wherein each individually selected Ht is a group of formula
wherein
n2 is 0, 1, 2, or 3;
n3 is 0, 1, 2, or 3;
Y is O or CH 2 ; and,
a wavy line indicates a point of bonding.