Oligomers
Molecules are provided for inducing or facilitating exon skipping in forming spliced mRNA products from pre-mRNA molecules in cells. The molecules may be provided directly as oligonucleotides or expression products of vectors that are administered to a subject. High rates of skipping can be achieved. High rates of skipping reduce the severity of a disease like Duchene Muscular Dystrophy so that the disease is more like Becker Muscular Dystrophy. This is a severe reduction in symptom severity and mortality.
1. A vector for ameliorating Duchenne muscular dystrophy the vector encoding an oligomer comprising at least 25 contiguous bases of a base sequence selected from the group consisting of:
(SEQ ID NO: 7)
a) CXX XXA GXX GCX GCX CXX XXC CAG GXX CAA;
(SEQ ID NO: 8)
b) CXX XXC XXX XAG XXG CXG CXC XXX XCC AGG;
and
(SEQ ID NO: 9)
c) XXA GXX GCX GCX CXX XXC CAG GXX CAA GXG;
or the oligomer comprising at least 28 contiguous
bases of the base sequence:
(SEQ ID NO: 6)
d) XGC XGC XCX XXX CCA GGX XCA AGX GGG AXA;
wherein X=U or T, wherein the base sequence of the oligomer can vary from the above sequence at up to two base positions, and wherein the oligomer is expressed when the vector is introduced into a human cell.
2. A pharmaceutical composition for ameliorating Duchenne muscular dystrophy, the composition comprising the vector according to claim 1 or the oligonucleotide encoded by the vector according to claim 1 , and a pharmaceutically acceptable carrier, adjuvant or vehicle.
3. A pharmaceutical composition according to claim 2 comprising a plurality of vectors encoding oligomers including the vector according to claim 1 and/or a plurality of oligomers including the oligomer encoded by the vector according to claim 1 wherein each oligomer encoded by a vector in the plurality of vectors causes skipping of an exon, and each oligomer in the plurality of oligomers causes skipping of an exon.
4. A method of ameliorating Duchenne muscular dystrophy in a human patient, the method comprising: administering a therapeutically effective amount of the vector according to claim 1 or the oligonucleotide encoded by the vector according to claim 1 to the patient such that the Duchenne muscular dystrophy is ameliorated.
5. The method according to claim 4 comprising administering a therapeutically effective amount of a plurality of vectors encoding oligomers including the vector according to claim 1 and/or a plurality of oligomers including the oligomer encoded by the vector according to claim 1 to the patient, wherein each oligomer encoded by a vector in the plurality of vectors causes skipping of an exon, and each oligomer in the plurality of oligomers causes skipping of an exon.
6. The vector according to claim 1 , wherein the oligomer causes exon skipping of exon 46 of the dystrophin gene.
7. The vector according to claim 6 , wherein the oligomer causes exon skipping in at least 50% of the total pre-mRNA.
8. The vector according to claim 1 , wherein the oligomer is between 25 and 35 bases in length.
9. The vector according to claim 1 , wherein the oligomer is 30 bases in length.
10. The pharmaceutical composition according to claim 2 , wherein the oligomer is conjugated to or complexed with a targeting protein.
11. The pharmaceutical composition according to claim 2 , wherein the oligomer is conjugated to or complexed with a drug for treating Duchenne muscular dystrophy.
12. The pharmaceutical composition according to claim 2 , wherein the oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.
13. The pharmaceutical composition according to claim 3 , wherein each oligomer encoded by a vector in the plurality of vectors causes skipping of a different exon, and each oligomer in the plurality of oligomers causes skipping of a different exon.