- Number
- 20170152261
- Published
- 2017-06-01
- Filed
- 2017-01-23
- Assignee
- Disney; Matthew D.
- Inventors
- Disney; Matthew D. et al.
- CPC
- A61K31/475; A61K47/65; C07D519/00; C07D471/04
- Verdict
- Set aside small molecules targeting RNA sequences - false-positive name match, unrelated Matthew D. Disney
- Source
- Google Patents · FreePatentsOnline
Abstract
The invention provides a series of bioactive smallmolecules that target expanded r(CGG) repeats, termed r(CGG).sup.exp, that causes FragileX-associated Tremor Ataxia Syndrome (FXTAS). The compound was identified by using information on the chemotypes and RNA motifs that interact. Specifically. 9-hydroxy-5,11-dimethyl-2-(2-(piperidin-1-yl)ethyl)-6H-pyrido[4,3-b]carbazol-2-ium, binds the 5′CGG/3′GGC motifs in r(CGG).sup.exp and disrupts a toxic r(CGG).sup.exp-protein complex. Specifically, dimeric compounds incorporating two 9-hydroxyellipticine analog structures can even more potently bind the 5′CGG/3′GGC motifs in r(CGG).sup.exp and disrupts a toxic r(CGG).sup.exp-protein complex. Structure-activity relationships (SAR) studies determined that the alkylated pyridyl and phenolic side chains are important chemotypes that drive molecular recognition of r(CGG) repeats, such as r(CGG).sup.exp. Importantly, the compound is efficacious in FXTAS model cellular systems as evidenced by its ability to improve FXTAS-associated pre-mRNAsplicing defects and to reduce the size and number of r(CGG).sup.exp-protein aggregates.
Background
BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 shows a schematic of the pathogenic mechanism in FXTAS.
FIG. 2 shows a schematic strategy in treatment of FXTAS.
FIG. 3 shows a schematic of the protein displacement assay that was used to identify small molecule inhibitors of the r(CGG).sub.12-DGCR8Δ interaction and to determine their potencies. The r(CGG).sub.12 oligonucleotide is labeled with a 5′-biotin while DGCR8Δ (blue cloud) contains a histidine (His) tag. Left, in the absence of inhibitor, DGCR8Δ binds to r(CGG).sub.12. Binding is quantified by using two antibodies that form a FRET pair—an anti-His antibody labeled with Tb that binds to DGCR8Δ and streptavidin labeled with XL665 that binds to r(CGG).sub.12. The two fluorophores are within close enough proximity to form a FRET pair. Tb is excited at 345 nm; the resulting emission (˜545 nm) excites XL665, which emits at 665 nm. Right, in the presence of inhibitor, the r(CGG).sub.12-DGCR8Δ interaction is disrupted, andthe two fluorophores are not within close enough proximity to form a FRET pair. Therefore, emission is only observed at 545 nm (due to Tb). XL665 emission is not observed.
FIG. 4 shows the structures of the small molecules identified from an RNA-focused library that inhibit the r(CGG).sub.12-DGCR8Δ interaction and derivatives of the most potent monomeric compound (1a). 1b-1f were used to construct structure-activity relationships and define the active pharmacopbore. Inhibition is markedly dec
Claims
1. A compound of formula (I) ##STR00028## wherein R.sup.1 is H, (C1-C6)alkyl, or (C1-C6)alkanoyl; R.sup.3 and R.sup.4 are independently H, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)alkoxyalkyl (C1-C6)haloalkoxyalkyl, or (C6-C10)aryl; R.sup.3 is (C1-C6)alkyl, (C1-C6)alkoxy(C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, aryl(C1-C6)alkyl, heterocyclyl(C1-C6)alkyl, heteroaryl(C1-C6)alkyl, or (R.sup.6).sub.2N—(C1-C6)alkyl, wherein R.sup.6 is H or (C1-C6)alkyl; wherein any alkyl, alkanoyl, alkoxy, aryl, heterocyclyl, or heteroaryl group can be substituted with 0-3 J groups, wherein J is any of halo, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)haloakyl, hydroxy(C1-C6)alkyl, alkoxy(C1-C6)alkyl, (C1-C6)alkanoyl, (C1-C6)alkanoyloxy, cyano, nitro, azido, R.sub.2N, R.sub.2NC(O), R.sub.2NC(O)O, R.sub.2NC(O)NR, (C1-C6)alkenyl, (C1-C6)alkynyl, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aroyl, (C6-C10)aryl(C1-C6)alkyl, (C6-C10)aryl(C1-C6)alkoxy, (C6-C10)aryloxy(C1-C6)alkyl, (C6-C10)aryloxy(C1-C6)alkoxy, (3- to 9-membered)heterocyclyl, (3- to 9-membered)heterocyclyl(C1-C6)alkyl, (3- to 9-membered)heterocyclyl(C1-C6)alkoxy, (5- to 10-membered)heteroaryl, (5- to 10-membered)heteroaryl(C1-C6)alkyl, (5- to 10-membered)heteroaryl(C1-C6)alkoxy, or (5- to 10-membered)heteroaroyl; R is independently at each occurrence H, (C1-C6)alkyl, or (C6-C10)aryl, wherein any alkyl or aryl group is substituted with 0-3 J; or, a dimeric r(CGG) binding compound of formula (II) ##STR00029## wherein R.sup.1, R.sup.1, R.sup.3, and R.sup.4 are as defined for the monomeric compound of formula (I), and wherein L is a linker comprising a polypeptide backbone bonded by two respective nitrogen atoms thereof to a nitrogen atom of arespective 1,2,3-triazole group via a respective (C1-C6)alkylene group optionally furthercomprising a glycyl residue, each respective triazole group being bonded via a (C1-C6)alkylene group to the respective pyridinium nitrogen atom of each ellipticine scaffold; or a pharmaceutically acceptable salt thereof; provided the compound of formula (I) is not any of ##STR00030## or a pharmaceutically acceptable salt thereof.
7. A pharmaceutical composition comprising a compound of any one of claims 1-6 and a pharmaceutically acceptable excipient.
8. A method of inhibiting a messenger RNA molecule with an repeat r(CGG) sequence from binding to a protein with a binding affinity for a RNA hairpin loop comprising a non-Watson-Crick 0-0 nucleotide pair, comprising contacting the messenger RNA molecule having the repeat r(CGG) sequence and an effective amount or concentration of a compound of formula (I) ##STR00032## wherein R.sup.1 is H, (C1-C6)alkyl, or (C1-C6)alkanoyl; R.sup.2 is H, (C1-C6)alkyl, or (C1-C6)alkanoyl; R.sup.3 and R.sup.4 are independently H, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)alkoxyalkyl (C1-C6)haloalkoxyalkyl, or (C6-C10)aryl; R.sup.5 is (C1-C6)haloalkoxy, aryl(C1-C6)alkyl, heterocyclyl(C1-C6)alkyl, heteroaryl(C1-C6)alkyl, or (R.sup.6).sub.2N—(C1-C6)alkyl, wherein R.sup.6 is H or (C1-C6)alkyl; wherein any alkyl, alkanoyl, alkoxy, aryl, heterocyclyl, or heteroaryl group can be substituted with 0-3 J groups,wherein J is any of halo, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)haloalkyl, hydroxy(C1-C6)alkyl, alkoxy(C1-C6)alkyl, (C1-C6)alkanoyl, (C1-C6)alkanoyloxy, cyano, nitro, azido, R.sub.2N, R.sub.2NC(O), R.sub.2NC(O)O, R.sub.2NC(O)NR, (C1-C6)alkenyl, (C1-C6)alkynyl, (C6-C10)aryl, (C6-C10)aryloxy, (C6-CJ O)aroyl, (C6-C10)aryl(C1-C6)alkyl, (C6-C10)aryl(C1-C6)alkoxy, (C6-C10)aryloxy(C1-C6)alkyl, (C6-C10)aryloxy(C1-C6)alkoxy, (3- to 9-membered)heterocyclyl,(3- to 9-membered)heterocycyl(C1-C6)alkyl, (3- to 9-membered)heterocyclyl(C1-C6)alkoxy, (5- to 10-membered)heteroaryl, (5- to 10-membered)heteroaryl(C1-C6)alkyl, (5- to 1-membered)heteroaryl(C1-C6)alkoxy, or (5- to 10-membered)heteroaroyl; R is independently at each occurrence H, (C1-C6)alkyl, or (C6-C10)aryl, wherein any alkyl or aryl group is substituted with 0-3 J; or, an effective amount or concentration of a dimeric r(CGG) binding compound of formula (II) ##STR00033## wherein R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are as definedfor the monomeric compound of formula (I), and wherein L is a linker comprising a polypeptide backbone bonded by two respective nitrogen atoms thereof to a nitrogen atom of a respective 1,2,3-triazole group via a respective (C1-C6)alkylene group optionally further comprising a glycyl residue, each respective triazole group being bonded via a (C1-C6)alkylenegroup to the respective pyridinium nitrogen atom of each ellipticine scaffold; or a pharmaceutically acceptable salt thereof; or a pharmaceutically acceptable salt thereof.
19. A method of treatment of Fragile X-associated Tremor Ataxia Syndrome, comprising administering to a patient afflicted therewith a therapeutically effective dose of a compound of formula (I) ##STR00037## wherein R.sup.1 is H, (C1-C6)alkyl, or (C1-C6)alkanoyl; R.sup.2 is H,(C1-C6)alkyl, or (C1-C6)alkanoyl; R.sup.3 and R.sup.4 are independently H, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)alkoxyalkyl (C1-C6)haloalkoxyalkyl, or (C6-C10)aryl; R.sup.5 is (C1-C6)alkyl, (C1-C6)alkoxy(C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, aryl(C1-C6)alkyl, heterocyclyl(C1-C6)alkyl, heteroaryl(C1-C6)alkyl, or (R.sup.6).sub.2N—(C1-C6)alkyl, wherein R.sup.1 is H or (C1-C6)alkyl; wherein any alkyl, alkanoyl, alkoxy, aryl, heterocyclyl, or heteroaryl group can be substituted with 0-3 J groups, wherein J is any of halo, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)haloalkyl, hydroxy(C1-C6)alkyl, alkoxy(C1-C6)alkyl, (C1-C6)alkanoyl, (C1-C6)alkanoyloxy, cyano, nitro, azido, R.sub.2N, R.sub.2NC(O), R.sub.2NC(O)O, R.sub.2NC(O)NR, (C1-C6)alkenyl, (C1-C6)alkynyl, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aroyl, (C6-C10)aryl(C1-C6)alkyl, (C6-C10)aryl(C1-C6)alkoxy, (C6-C10)aryloxy(C1-C6)alkyl (C6-C10)aryloxy(C1-C6)alkoxy, (3- to 9-membered)heterocyclyl, (3- to 9-membered)heterocyclyl(C1-C6)alkyl, (3- to 9-membered)heterocyclyl(C1-C6)alkoxy, (5- to 10-membered)heteroaryl, (5- to 10-membered)heteroaryl(C1-C6)alkyl, (5- to 10-membered)heteroaryl(C1-C6)alkoxy, or (5- to 10-membered)heteroaryl; R is independently at each occurrence H, (C1-C6)alkyl, or (C6-C10)aryl, wherein any alkyl or aryl group is substituted with 0-3 J; or, an effective amount or concentration of a dimeric r(CGG) binding compound of formula (II) ##STR00038## wherein R.sup.1, R.sup.2, R.sup.1, and R′ are as defined for the monomeric compound of formula (I), and wherein L is a linker comprising polypeptide backbone bonded by two respective nitrogen atoms thereof to a nitrogen atom of a respective 1,2,3-triazole group via a respective (C1-C6)alkylene group optionally further comprising a glycyl residue, each respective triazole group being bonded via a (C1-C6)alkylene group to the respective pyridinium nitrogen atom of each ellipticine scaffold; or a pharmaceutically acceptable salt thereof; or a pharmaceutically acceptable salt thereof.
27. A method of inhibiting a messenger RNA molecule with a repeat r(CGG) sequence from binding to a protein with a binding affinity for a RNA hairpin loop comprising a non-Watson-Crick G-G nucleotide pair, comprising contacting the messenger RNA molecule having the repeat r(CGG) sequence and an effective amount or concentration of 9-hydroxyellipticine bearing an N-substituted pyridinium moiety, or ananalog thereof, or a dimeric derivative of 9-hydroxyellipticine wherein two 9-hydroxyellipticine scaffolds are linked via a linker group.
29. A method of treatment of Fragile X-associated Tremor Ataxia Syndrome, comprising administering to a patient afflicted therewitha therapeutically effective dose of 9-hydroxyellipticine comprising an N-substituted pyridinium moiety, or an analog thereof, or a dimeric derivative of 9-hydroxyellipticine wherein two 9-hydroxyellipticine scaffolds are linked via a linker group.