Complexes comprising an anti-transferrin receptor antibody linked to an oligonucleotide
Inventors
Subramanian, Romesh R. • Qatanani, Mohammed T. • Weeden, Timothy • Desjardins, Cody A.
Assignees
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Publication Number
US-12173078-B2
Publication Date
2024-12-24
Expiration Date
Abstract
Aspects of the disclosure relate to complexes comprising a muscle-targeting agent covalently linked to a molecular payload. In some embodiments, the muscle-targeting agent specifically binds to an internalizing cell surface receptor on muscle cells. In some embodiments, the molecular payload inhibits expression or activity of a DMPK allele comprising a disease-associated-repeat. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.
Core Innovation
Muscle-targeting complexes and molecular payloads are provided for delivering an oligonucleotide payload to muscle cells, in particular for treating myotonic dystrophy. The disclosed complexes include an anti-transferrin receptor antibody covalently linked to at least one oligonucleotide, and the disclosure also relates to muscle-targeting agents including ENT2 substrates, OCTN2 substrates, and soluble proteins such as hemojuvelin that are linked to molecular payloads.
The oligonucleotide comprises one or more modifications and includes a region of complementarity of at least 15 nucleotides in length to a nucleotide sequence as set forth in SEQ ID NO: 15, with the oligonucleotide being in the range of 15-20 nucleotides in length. The oligonucleotide further includes a gap region flanked at both the 5′ and 3′ by flanking regions, where each flanking region independently comprises one or more 2′-modified nucleosides, the nucleosides of the gap region are 2′-deoxyribonucleosides, and the oligonucleotide comprises one or more phosphorothioate internucleoside linkages.
The disclosure also includes DMPK-targeting oligonucleotide payloads and their formats, including gapmer, siRNA, and other formats, with sequence identifiers for human and mouse DMPK regions and related complementary sequences. Linker chemistry connects targeting agents to payloads through cleavable and non-cleavable linkers, including Val-Cit protease-sensitive linkers, and example constructs include transferrin receptor antibody–oligonucleotide conjugates that use a Val-Cit linker.
A corresponding composition is disclosed that includes complexes comprising the same antibody and oligonucleotide architecture, and the disclosure further addresses antibody–oligonucleotide conjugate design considerations such as drug-to-antibody ratio. Experimental results are described for DMPK knockdown in cell models, mouse tissue models, and non-human primate contexts, together with formulations and in vivo treatment/testing for myotonic dystrophy (DM1).
Claims Coverage
The independent claims are directed to a muscle-targeting complex comprising a specified anti-TfR1 antibody covalently linked to a defined, modified oligonucleotide with complementarity to SEQ ID NO: 15, and to a composition comprising such complexes. Across the independent claims, the inventive coverage is organized around the antibody binding region constraint, the oligonucleotide length/complementarity constraint to SEQ ID NO: 15, and the specific oligonucleotide gap/flanking-region and chemistry constraints.
Anti-TfR1 antibody binds a specified residue range but not the transferrin binding site
An anti-transferrin receptor antibody that binds in the range of C89 to F760 of human transferrin receptor protein 1 (TfR1) having an amino acid sequence as set forth in SEQ ID NO: 1, wherein the anti-transferrin receptor antibody does not specifically bind to the transferrin binding site of TfR1.
Oligonucleotide with ≥15-nt complementarity to SEQ ID NO: 15 and 15-20-nt length
An oligonucleotide that comprises one or more modifications and comprises a region of complementarity of at least 15 nucleotides in length to the nucleotide sequence as set forth in SEQ ID NO: 15, wherein the oligonucleotide is in the range of 15-20 nucleotides in length.
Gap/flanking architecture with 2′-modified flanks and 2′-deoxyribonucleoside gap plus phosphorothioate linkages
An oligonucleotide comprising a gap region flanked both 5′ and 3′ by flanking regions, wherein each flanking region independently comprises one or more 2′-modified nucleosides selected from the group consisting of a 2′-O-methoxyethyl nucleoside, a 2′,4′-bridged nucleoside, and combinations thereof, and wherein each nucleoside of the gap region is a 2′-deoxyribonucleoside, and wherein the oligonucleotide comprises one or more phosphorothioate internucleoside linkages.
Composition comprising the antibody-oligonucleotide complexes
A composition comprising complexes that comprise an anti-transferrin receptor antibody covalently linked to at least one oligonucleotide, wherein the antibody binds in the range of C89 to F760 of human TfR1 having an amino acid sequence as set forth in SEQ ID NO: 1 and does not specifically bind to the transferrin binding site of TfR1, and wherein the oligonucleotide and its gap/flanking architecture and phosphorothioate internucleoside linkages follow the same constraints as recited for the complex.
Across the independent claims, protection centers on a covalent antibody–oligonucleotide complex and compositions of such complexes in which the anti-TfR1 antibody binds a defined TfR1 residue range without specifically binding the transferrin binding site, and the oligonucleotide includes one or more modifications with a complementarity region to SEQ ID NO: 15 and a 15-20 nucleotide length, together with a defined gap and flanking-region nucleoside pattern and phosphorothioate internucleoside linkages.
Stated Advantages
Muscle-targeted DMPK knockdown is described across muscle-related tissues with minimal effect in non-muscle tissues.
DMPK reduction is described as persistent up to 28 days.
The document describes dose dependence of DMPK reduction.
Safety/tolerability readouts in cynomolgus monkeys are described.
Muscle targeting selectivity.
DMPK allele expression/activity inhibition in muscle cells.
Durability of the effect.
Tolerability/safety in mice and cynomolgus monkeys.
DMPK knockdown is reported in cell models, mouse tissue models, and non-human primates.
The disclosure provides multiple muscle-targeting agent types and multiple payload options.
Cleavable Val-Cit protease-sensitive linkers are described as an option for linking targeting agents to payloads.
Documented Applications
Use in muscle-related tissues for DMPK mRNA reduction, including application contexts related to myotonic dystrophy type 1 (DM1) and the DMPK 3′ non-coding region CTG trinucleotide repeat.
In vivo reduction of DMPK in mouse tissues including gastrocnemius, heart, esophagus, tibialis anterior, and soleus as described in the document.
In vivo and in vitro evaluation of DMPK knockdown driven by the antibody–oligonucleotide conjugate in stated cell and animal contexts, with safety/tolerability readouts in cynomolgus monkeys.
Treating myotonic dystrophy using muscle-targeting complexes that deliver oligonucleotide payloads to muscle cells for inhibition of DMPK allele expression/activity.
Use in muscle tissue targeting and assessment of tissue selectivity, including durability and tolerability/safety evaluation in mice and cynomolgus monkey muscle tissue.
Treatment/testing in vivo for myotonic dystrophy (DM1) using the described targeting agent/oligonucleotide constructs.
DMPK knockdown in cell and mouse tissue models.
DMPK knockdown in non-human primate contexts.
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