Nucleic acid binding compounds and methods of use
The present invention relates to homo- and hetero-dimer compounds formed by a disulfide, sulfinyl thio, or olefin bond between two monomers. A method of making a homo- or hetero-dimer compound is also disclosed. The present invention also relates to monomer compounds capable of forming homo- or hetero-dimer compounds, as well as oligomers formed via linkage of one or more dimers. Also disclosed are methods of inhibiting the activity of target RNA molecules, particularly those having a secondary structure that include a stem or stem-loop formation. Dimer compounds capable of inhibiting the activity of an HIV-I RNA frameshifting stem-loop and a (CUG)n expanded repeat stem-loop are disclosed, as are methods of treating diseases associated with these target RNA molecules. The dimer compounds can also be used for selectively detecting presence of the target RNA molecule in a sample.
1. A homo- or hetero-dimer compound of two monomers having a structure
wherein, for each monomer (I)
Q is NH 2 ;
n is independently an integer from 0 to about 5;
Z is independently a tripeptide or tetrapeptide comprising Phe-Pro-allyl-Gly, Val-His-allyl-Gly, Lys-allyl-Gly-Pro, or Lys-allyl-Gly-Asn, where one of the amino acids is linked to an amino acid in the other monomer by an olefin bond; and
A is selected from
wherein R 2 and R 3 are independently selected from the group consisting of hydrogen, methyl, ethyl, propyl, amino, methylamine, ethylamine, dimethylamine, diethylamine, methoxy, ethoxy, propoxy, hydroxyl, cyano, and thiocyanate and A is connected to Z via a carbonyl linkage.
2. The dimer compound according to claim 1 , wherein an olefin bond links together two monomers having a structure
wherein, for each monomer (II)
Q is NH 2 ;
n is independently an integer from 0 to about 5; and
Xaa is allyl-Gly.
3. The dimer compound according to claim 1 , wherein an olefin bond links together a first monomer having a structure
and a second monomer having a structure
wherein, for each of monomers (II) and (III),
Q is NH 2 ;
n is independently an integer from 0 to about 5; and
Xaa is allyl-Gly.
4. The dimer compound according to claim 1 , wherein an olefin bond links together a first and a second monomer, wherein the first and second monomers independently have the structure
where, for each of monomers (V) and (VI),
Q is NH 2 ;
n is independently an integer from 0 to about 5;
Xaa 1 is allyl-Gly; and
Xaa 2 is Pro or Asn.
5. A method of inhibiting HIV-1 proliferation, said method comprising:
providing a dimer compound according to claim 1 ; and
contacting an HIV-1 mRNA that encodes Pol polyprotein with the dimer compound under conditions effective to alter normal expression of the Pol polyprotein and thereby inhibit HIV-1 proliferation.
6. A method of treating HIV-1 in a human patient, said method comprising:
administering to a human patient a dimer compound according to claim 1 under conditions effective to alter normal expression of HIV-1 Pol polyprotein, thereby disrupting HIV-1 proliferation to treat the human patient for HIV-1.
7. A composition comprising a homo- or hetero-dimer compound according to claim 1 and a carrier.
8. The composition according to claim 7 , wherein the carrier is a pharmaceutically-acceptable carrier.
9. The composition according to claim 8 , wherein the carrier is selected from a polyethylene glycol conjugate, a liposome, oligoarginine, and a nanoparticle carrier.
10. A method of making a homo- or hetero-dimer compound, said method comprising:
providing a first and second monomer having a structure
wherein:
Q is NH 2 ;
n is independently an integer from 0 to about 5;
Z is independently a tripeptide or tetrapeptide comprising Phe-Pro-allyl-Gly, Val-His-allyl-Gly, Lys-allyl-Gly-Pro, or Lys-allyl-Gly-Asn; and
A is selected from
wherein R 2 and R 3 are independently selected from the group consisting of hydrogen, methyl, ethyl, propyl, amino, methylamine, ethylamine, dimethylamine, diethylamine, methoxy, ethoxy, propoxy, hydroxyl, cyano, and thiocyanate and A is connected to Z via a carbonyl linkage; and
reacting the first and second monomers under conditions effective to form an olefin bond, thereby forming the homo- or hetero-dimer compound according to claim 1 .
11. A method of altering the activity of a target RNA molecule comprising:
contacting the RNA molecule with a dimer compound according to claim 1 that selectively binds to the target RNA molecule, said contacting being effective to alter activity of the RNA molecule.
12. The method according to claim 11 wherein the target RNA molecule is an RNA molecule of a pathogen, and the activity of the RNA molecule is critical to survival and/or proliferation of the pathogen.
13. The method according to claim 11 wherein the target RNA molecule is an RNA molecule of a mammal and comprises an expanded repeat sequence, and the activity of the RNA molecule is implicated in a disease state of the mammal.
14. The method according to claim 11 wherein the target RNA molecule contains a frameshift site or an expanded nucleotide repeat.
15. The method according to claim 11 , wherein the frameshift site is −1 ribosomal frameshifting of SARS, HIV-1, Hepatitis C virus, Rous Sarcoma Virus, human T-Cell Leukemia Virus Type II, and Coronavirus.
16. A method of treating a subject for a disorder caused by an expanded RNA repeat sequence, said method comprising:
administering to the subject a dimer compound according to claim 1 under conditions effective to alter function of an expanded RNA repeat sequence, thereby disrupting interaction between the RNA repeat sequence and splicing proteins to treat the subject for the disorder.
17. The method according to claim 16 , wherein the expanded repeat sequence is selected from the group of (CUG) n , (GAA) n , (CAG) n , (CGG) n , (CCG) n , (CCTG) n , (ACAGGGGT(G/C)) n , (CCCCGCCCCGCG) n , and (ATTCT) n .
18. The method according to claim 16 , wherein the disorder is selected from the group consisting of inherited neuropathy, muscular dystrophy, Friedreich ataxia, lysosomal storage disease, mitochondrial disorder, Huntington's disease, spinocerebellar ataxia (Machado-Joseph disease), dentatorubral pallidoluysian atrophy, spinobulbar muscular atrophy (Kennedy's disease), fragile X syndrome, Jacobsen syndrome, diabetes mellitus, and myoclonus epilepsy.