Nanoparticle type oligonucleotide structure having high efficiency and method for preparing same
Inventors
Park, Han Oh • Chae, Jeiwook • Yoon, Pyoung Oh • Han, Boram • Choi, Gi-Eun • Ko, Youngho • Kwon, Taewoo • Lee, Jae Don • Kim, Sun Gi
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Assignees
Member
BioneerBioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Bioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Publication Number
US-10030243-B2
Publication Date
2018-07-24
Expiration Date
Abstract
The present invention relates to an oligonucleotide structure and a method for preparing the same and, more particularly, to an oligonucleotide structure in which a polymer compound is linked to an oligonucleotide via a covalent bond to improve in vivo stability of the oligonucleotide and cellular delivery efficiency of the oligonucleotide; and to a method for preparing the same. The oligonucleotide structure is improved into a homogenous material, thereby solving the problem in material verification due to polydispersion characteristics occurring when a hydrophilic material linked to the oligonucleotide is a synthetic polymer; the oligonucleotide structure is easy to synthesize compared with the existing process; and the size of a double-stranded oligo RNA structure can be accurately adjusted through the control of the repetition number of a hydrophilic material block, and thus, the gene expression regulation function of the oligonucleotide does not deteriorate through the synthesis of the optimized oligonucleotide structure, and the oligonucleotide can be delivered into cells at even a relatively low-concentration dosage. Therefore, the oligonucleotide structure of the present invention can be useful as a novel type oligonucleotide delivery system as well as a tool for treating cancers, infectious diseases, and the like.
Core Innovation
The invention relates to an oligonucleotide structure represented by Structural Formula (1) or Structural Formula (2). The structure includes a hydrophilic material monomer (A) and a hydrophobic material (B), with a linker (J) connecting m hydrophilic material monomers, and with X and Y representing simple covalent bonds or linker-mediated covalent bonds. A region (R) is provided as a single-stranded or double-stranded oligonucleotide, and m is an integer of 1 to 15 while n is an integer of 1 to 10, thereby defining a modular architecture combining hydrophilic blocks, hydrophobic material, and an oligonucleotide region.
The oligonucleotide structure further defines Q as either (LiZj) or P-J1-J2, where L is a ligand specifically bonded to a receptor that promotes target cell internalization through receptor-mediated endocytosis (RME). Z is a linker that mediates a simple covalent bond or a bond between the hydrophilic material block monomer and the ligand, and i and j define integer parameters for the presence and bonding arrangement. P is an amine group or a polyhistidine group, and J1 and J2 are independently linkers that mediate a simple covalent bond or connect the amine or polyhistidine group with the hydrophilic material.
The patent describes embodiments in which these modular amphipathic oligonucleotide structures are used to synthesize single-stranded and double-stranded SAMiRNA on solid support, including structures that include a hydrophilic monomer block and a hydrophobic disulfide-containing material. In examples directed to Survivin-targeting dsRNA, ligand-functionalized nanoparticles are described as thermodynamically stable, with hydrophilic blocks oriented outward and a hydrophobic core inward. Nanoparticles are characterized by critical micelle concentration (CMC) and physical measurements such as size and polydispersity index (PDI), and the resulting formulations are described as showing gene-expression inhibition in HeLa cells.
Claims Coverage
The partial content provides one independent claim (clm-00001). This claim defines the core modular oligonucleotide architecture with a ligand specifically enabling receptor-mediated endocytosis (RME) and includes the option to incorporate an amine group or polyhistidine group into the connector framework.
Modular amphipathic oligonucleotide structure with hydrophilic and hydrophobic blocks
An oligonucleotide structure having a structure represented by Structural Formula (1) or Structural Formula (2), wherein A is a hydrophilic material monomer, B is a hydrophobic material, J connects m hydrophilic material monomers, X and Y are simple covalent bonds or linker-mediated covalent bonds, R is single-stranded or double-stranded oligonucleotide, m is an integer of 1 to 15, and n is an integer of 1 to 10.
Receptor-mediated endocytosis ligand incorporated via Q, L, and Z
The structure wherein Q is (LiZj) or P-J1-J2, L is a ligand specifically bonded to a receptor that promotes target cell internalization through receptor-mediated endocytosis (RME), Z is a linker that mediates a simple covalent bond or a bond between the hydrophilic material block monomer and the ligand, and i and j define integer parameters of 0 to 5 and 0 or 1 with the condition that when i is 0, j is necessarily 0.
Amine or polyhistidine functionality integrated via P, J1, and J2
The structure wherein P means an amine group or a polyhistidine group, and J1 and J2 are independently linkers that mediate a simple covalent bond, or a bond between the amine group or the polyhistidine group with the hydrophilic material.
Overall, independent claim clm-00001 covers a modular amphipathic oligonucleotide architecture (Structural Formula (1) or (2)) that combines hydrophilic material monomer blocks and a hydrophobic material region with an oligonucleotide region, and that incorporates a receptor-mediated endocytosis (RME) ligand via Q, L, and Z. The claim also provides optional incorporation of amine or polyhistidine functionality through P, J1, and J2.
Stated Advantages
Improved in vivo stability and cellular delivery.
Reduced polydispersity versus PEG-based conjugates.
Easier material verification and size control through uniform hydrophilic block and controlled repeat numbers.
Improved reproducibility.
Improved targeting through ligand, including passive and active targeting described via EPR/passive/active targeting.
Reduced polydispersity-related quality-control issues.
Simplified synthesis/purification.
Easier material analysis.
Tunable nanoparticle size.
Improved delivery via RME ligands.
Enhanced endosomal escape/lysosomal resistance.
Documented Applications
Forming self-assembled nanoparticles (SAMiRNA) from the described oligonucleotide–polymer–lipid amphipathic architecture.
Promoting target cell internalization through receptor-mediated endocytosis (RME) using a ligand specifically bonded to a receptor.
Use for gene-expression inhibition, as described through survivin (Survivin) constructs and inhibition trends.
The document describes subsequent nanoparticle property assessment including CMC, size/PDI, and inhibition trends.
Survivin-targeting dsRNA SAMiRNA formulations and corresponding ligand-functionalized nanoparticles, including embodiments using ligand N-acetylgalactosamine (NAG), described as inhibiting gene expression in HeLa cells.
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