Minor groove binder (MGB)-oligonucleotide miRNA antagonists
Inventors
Khvorova, Anastasia • Vermeulen, Annaleen • Kaiser, Rob • Karpilow, Jon • Vermeulen, Nicolaas M. J. • Mahoney, Walt
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-8980855-B2
Publication Date
2015-03-17
Expiration Date
Abstract
Compositions and methods for inhibiting the actions of non-coding RNAs such as miRNAs and piRNAs are provided. The compositions comprise single or double stranded oligonucleotides conjugated with Minor Groove Binders (“MGBs”). The oligonucleotides can vary in length, can contain nucleotides having one or more modifications, and have regions that are substantially complementary to one or more mature miRNAs or piRNAs.
Core Innovation
The invention provides an inhibitor composition for inhibiting miRNA activity in vitro or in vivo. The inhibitor composition comprises an oligonucleotide and a minor groove binder (MGB), wherein the MGB is conjugated to the 5′ end of the oligonucleotide. The oligonucleotide comprises nucleotides in which all nucleotides comprise ribofuranose having a structure in which Rz is —OCH3, and in which B is a normal base or a modified base.
The inhibitor composition is configured for treatment of a condition characterized by over-expression of miRNA by administering the inhibitor composition to a subject at a concentration sufficient to inhibit the action of the miRNA. The oligonucleotide nucleotides are linked by natural phosphodiester linkages. The design includes options for modified bases or universal bases, and the MGB can be constructed from selected MGB structures with defined substituent and linkage options.
The described embodiments include MGB conjugates in which a linker may connect the MGB to the oligonucleotide, and structural parameters for the inhibitor composition are parameterized in detail. In at least one example, dual-luciferase psiCHECK-2 reporter assays are used to evaluate inhibitor function, and the document reports that MGB placement and oligonucleotide chemistry substantially improve inhibitory potency versus 2′-O-methyl controls for let-7c and miR-21.
Claims Coverage
The independent claims cover methods of inhibiting miRNA activity in vitro or in vivo and methods of treating conditions characterized by miRNA over-expression, with an additional independent claim specifying single-stranded RNA oligonucleotide inhibitors conjugated to CDPI3 for let-7 or miR-21. Across the independent claims, the core coverage includes an inhibitor composition comprising an oligonucleotide with ribofuranose (Rz=—OCH3) linked by natural phosphodiester linkages and an MGB conjugated to the 5′ end.
Method of inhibiting miRNA activity using 5′-end MGB-conjugated ribofuranose oligonucleotide
A method of inhibiting miRNA activity in vitro or in vivo by introducing an inhibitor composition to a location where miRNA activity exists; the inhibitor composition comprises an oligonucleotide and an MGB, wherein all nucleotides comprise ribofuranose having Rz=—OCH3, B is a normal base or a modified base, nucleotides are linked by natural phosphodiester linkages, and the MGB is conjugated to the 5′ end of the oligonucleotide.
Method of treating miRNA over-expression using 5′-end MGB-conjugated ribofuranose oligonucleotide
A method of treating a condition characterized by over-expression of miRNA by administering an inhibitor composition to a subject at a concentration sufficient to inhibit the action of said miRNA; the inhibitor composition comprises an oligonucleotide and an MGB, wherein the oligonucleotide comprises nucleotides in which all nucleotides comprise ribofuranose having Rz=—OCH3, B is a normal base or a modified base, nucleotides are linked by natural phosphodiester linkages, and the MGB is conjugated to the 5′ end of the oligonucleotide.
Treatment of let-7 or miR-21 using CDPI3-conjugated single-stranded RNA inhibitor
A method of treating a condition characterized by over-expression of miRNA by administering an inhibitor composition to a subject at a concentration sufficient to inhibit the action of said miRNA; the inhibitor composition comprises a single-stranded oligonucleotide consisting of RNA and an MGB, wherein all nucleotides comprise ribofuranose having Rz=—OCH3 and B is a normal base or a modified base, nucleotides are linked by natural phosphodiester linkages, the MGB is CDPI3 conjugated to the 5′ end of the oligonucleotide, and the miRNA is let-7 or miR-21.
The independent claims collectively require an inhibitor composition containing an oligonucleotide with ribofuranose (Rz=—OCH3) linked by natural phosphodiester linkages and an MGB conjugated to the 5′ end; the method of use is either introducing the inhibitor composition where miRNA activity exists or administering it to a subject for a condition characterized by miRNA over-expression, with an additional independent claim specifying single-stranded RNA and an MGB specified as CDPI3 for let-7 or miR-21.
Stated Advantages
Substantially improved inhibitory potency versus 2′-O-methyl controls for let-7c and miR-21, as reported in the document example.
Documented Applications
In vitro or in vivo modulation of miRNA activity by introducing an inhibitor composition to a location where miRNA activity exists.
In vitro/in vivo modulation and therapeutic/prophylactic use for diseases/conditions characterized by miRNA over-expression.
Interested in licensing this patent?