Compositions and methods for capping RNAS
Inventors
Cheng, Christopher • RAJEEV, Kallanthottathil • Reiss, Caroline
Assignees
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Publication Number
US-12264175-B2
Publication Date
2025-04-01
Expiration Date
Abstract
Provided herein are compositions and methods for preparation of 5′ end region-modified mRNAs. In particular, the instant disclosure relates to novel mRNA 5′ end region motifs and sequence initiators therefore together with assays that are capable of measuring the aspects of the functionality of those motifs and sequence initiators. Further provided herein are compositions and methods of treating conditions related to coronary disease.
Core Innovation
The invention relates to an in vitro-transcribed (IVT) mRNA sequence initiator comprising a compound of formula (II), or a salt or solvate thereof, and to an mRNA having a 5′-end region motif (I′). The nucleobases B1 through Bn are independently a natural, a modified, or an unnatural nucleobase, and the motifs are defined by multiple position-specific substituent selections at Z′, Z″, Z1, Z2, Z3, Z4, Zn, Q1 through Q4, X1 through X5 or Xn, Y1 through Y5 or Yn, and A, A1, and A2, with p being 0, 1, 2, 3, 4, 5, or 6.
Specific motif embodiments are disclosed, including Motif (I′-d), in which B2 is adenine and B3 is guanine. In that embodiment, Q1 and Q4 are CH2O, Q2 and Q3 are O, X1, X2, and X3 are OH or O, Y1, Y2, and Y3 are O, and A1 and A2 are O.
The description additionally states that the 5′-end motif or initiator can increase protein expression and reduce immune stimulation, including reduced RIG-I activation. It further describes integration with in vitro-transcription (IVT) processes and RNA embodiments including mRNA, guide RNA, and nuclease mRNA.
Claims Coverage
The consolidated claim coverage includes three inventive features: an IVT mRNA sequence initiator of formula (II), an mRNA having a 5′-end region motif (I′), and a narrower Motif (I′-d) embodiment. These claims center on parameterized nucleobase and substituent selections with p from 0 to 6.
In vitro-transcribed mRNA sequence initiator of formula (II)
An in vitro-transcribed (IVT) mRNA sequence initiator comprising a compound of formula (II) or a salt or solvate thereof, wherein B1 through Bn are independently a natural, a modified, or an unnatural nucleobase; substituent positions Z′, Z″, Z1, Z2, Z3, Z4, and Zn are independently selected from specified groups; Q1 through Q4, X1 through X5, Y1 through Y5, and A, A1, and A2 are independently selected from specified groups; and p is 0, 1, 2, 3, 4, 5, or 6.
mRNA having a 5′-end region motif (I′)
An mRNA having a 5′-end region motif (I′) wherein B1 through Bn are independently a natural, a modified, or an unnatural nucleobase; Z1 and Z2, Z3, Z4 and Zn, Q1 and Q4, Q2 and Q3, X1 through Xn, Y1 through Yn, and A, A1, and A2 are independently selected from specified groups; and p is 0, 1, 2, 3, 4, 5, or 6.
Specific Motif (I′-d) instantiation
The mRNA having a 5′-end region motif (I′) narrowed to Motif (I′-d), where B2 is adenine and B3 is guanine, Q1 and Q4 are CH2O, Q2 and Q3 are O, X1, X2, and X3 are OH or O, Y1, Y2, and Y3 are O, and A1 and A2 are O.
Together, the claims cover an IVT mRNA sequence initiator defined by formula (II) and an mRNA containing a 5′-end region motif (I′), each defined by independently selectable nucleobases and substituent sets across multiple labeled positions, with a narrower Motif (I′-d) embodiment specifying selected base identities and substituent assignments.
Stated Advantages
Increase protein expression.
Reduce immune stimulation, including reduced RIG-I activation.
Documented Applications
RNA embodiments including mRNA, guide RNA, and nuclease mRNA.
Delivery formulations using lipid nanoparticles.
Gene editing.
Coronary disease risk reduction.
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