Aptamer-mRNA conjugates for targeted protein or peptide expression and methods for their use

• US 10,272,165 B2
• Filed: 07/11/2016
• Issued: 04/30/2019
• Est. Priority Date: 06/12/2011
• Status: Active Grant

• Current Assignee: City of Hope
• Original Assignee: City of Hope
• Inventors: Rossi, John J., Bobbin, Maggie
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US15/207,280
• Publication Number:
  US 20170043040A1
• Time in Patent Office: 1,023 Days
• CPC Class Codes: A61K 38/465 acting on ester bonds (3.1), e.g. lipases, ribonucleases
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: A61K 49/12 Macromolecular compounds
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/1132 against retroviridae, e.g. HIV
• CPC Class Codes: C12N 15/115 Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/16 Aptamers
• CPC Class Codes: C12N 2310/3519 Fusion with another nucleic acid
• CPC Class Codes: C12N 2320/32 : Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12Y 301/21004 : Type II site-specific deoxyribonuclease (3.1.21.4)

Abstract

An aptamer-mRNA conjugate is provided. The aptamer-mRNA conjugate may include an aptamer component that binds a membrane associated protein on a target cell and an mRNA component that is expressed by the target cell.

Claims

1. An imaging method comprising:

administering an effective dose of one or more aptamer-mRNA conjugates to a subject, wherein each of the one or more aptamer-mRNA conjugates comprises;

an aptamer component that binds a membrane associated protein on a target cell; and

an mRNA molecule that encodes a diagnostic agent that is expressed by the target cell, wherein the aptamer component is conjugated to the mRNA molecule; and

exposing the subject to an imaging method.

2. The imaging method of claim 1, wherein the diagnostic agent that is expressed by the target cell is a fluorescent protein or peptide.

3. The imaging method of claim 2, wherein the fluorescent protein or peptide is green fluorescent protein (GFP), enhanced GFP (eGFP), red fluorescent protein, blue fluorescent protein, yellow fluorescent protein, cyan fluorescent protein, sapphire fluorescent protein, reef coral fluorescent protein, or a derivative thereof.

4. The imaging method of claim 1, wherein the diagnostic agent that is expressed by the target cell is a luminescent protein or peptide.

5. The imaging method of claim 4, wherein the luminescent protein or peptide is luciferase, aequorin, or a derivative thereof.

6. The imaging method of claim 1, wherein the aptamer binds a target molecule on the surface of a cell and the mRNA molecule is expressed in the cell and visualized by the imaging method.

7. The imaging method of claim 1, wherein the aptamer binds a target molecule on a pathogen and visualized by the imaging method.

8. The imaging method of claim 7, wherein the pathogen is human immunodeficiency virus (HIV).

9. The imaging method of claim 1, wherein the imaging method is positron emission tomography (PET), computer assisted tomography (CAT), single photon emission computerized tomography, x-ray, fluoroscopy, or magnetic resonance imaging (MRI).

10. The imaging method of claim 1, wherein the aptamer-mRNA conjugates are delivered via dendrimers.

11. The imaging method of claim 1, wherein the aptamer component and the mRNA molecule are conjugated covalently.

12. The imaging method of claim 1, wherein the aptamer component and the mRNA molecule are conjugated non-covalently.