Splice-region antisense composition and method
Antisense compositions targeted against an mRNA sequence coding for a selected protein, at a region having its 5′ end from 1 to about 25 base pairs downstream of a normal splice acceptor junction in the preprocessed mRNA, are disclosed. The antisense compound is RNase-inactive, and is preferably a phosphorodiamidate-linked morpholino oligonucleotide. Such targeting is effective to inhibit natural mRNA splice processing, produce splice variant mRNAs, and inhibit normal expression of the protein.
1. A pharmaceutical composition comprising an oligomer of 8 to 40 morpholino subunits connected by phosphorous-containing intersubunit linkages, wherein the 5′ terminal morpholino subunit comprises a group of the formula:
or a pharmaceutically acceptable salt thereof, wherein each R 1 is independently a C 1 -C 6 alkyl, and a pharmaceutically acceptable carrier.
2. The pharmaceutical composition of claim 1 , wherein the 5′ terminal morpholino subunit of the oligomer is of a formula:
wherein each P i is independently a purine or pyrimidine base-pairing moiety.
3. The pharmaceutical composition of claim 1 , wherein the oligomer is of a formula:
or a pharmaceutically acceptable salt thereof,
wherein:
each R 1 is independently a C 1 -C 6 alkyl;
each P i is independently a purine or pyrimidine base-pairing moiety;
X is an integer from 3 to 19; and
R 3 is H.
4. The pharmaceutical composition of claim 3 , wherein each R 1 is independently selected from methyl, ethyl, isopropyl, n-butyl, isobutyl, and t-butyl.
5. The pharmaceutical composition of claim 3 , wherein each R 1 is methyl.
6. The pharmaceutical composition of claim 3 , wherein each P i is independently selected from adenine, cytosine, guanine, uracil, and thymine.
7. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is formulated for intravenous administration.
8. The pharmaceutical composition of claim 1 , wherein the oligomer is of a formula:
or a pharmaceutically acceptable salt thereof,
wherein:
each R 1 is independently a C1-C6 alkyl;
each P i is independently a purine or pyrimidine base-pairing moiety;
X is an integer from 6 to 38; and
R 3 is H.
9. The pharmaceutical composition of claim 8 , wherein each R 1 is independently selected from methyl, ethyl, isopropyl, n-butyl, isobutyl, and t-butyl.
10. The pharmaceutical composition of claim 8 , wherein each R 1 is methyl.
11. The pharmaceutical composition of claim 8 , wherein each P i is independently selected from adenine, cytosine, guanine, uracil, and thymine.
12. A pharmaceutical composition comprising a compound of formula:
or a pharmaceutically acceptable salt thereof, wherein X is an integer from 3 to 19, and each P i is independently a purine or pyrimidine base-pairing moiety selected from adenine, cytosine, guanine, uracil, and thymine, and a pharmaceutically acceptable carrier.
13. A pharmaceutical composition comprising a compound of formula:
or a pharmaceutically acceptable salt thereof, wherein X is an integer from 6 to 38, and each P i is independently a purine or pyrimidine base-pairing moiety selected from adenine, cytosine, guanine, uracil, and thymine, and a pharmaceutically acceptable carrier.