DNA APTAMERS BINDING TO MOLECULAR TARGETED AGENTS AND DETECTION METHOD OF MOLECULAR TARGETED MEDICINE USING THE SAME

• US 20190284560A1
• Filed: 11/29/2016
• Published: 09/19/2019
• Est. Priority Date: 11/30/2015
• Status: Active Grant

• Current Assignee: Nissan Chemical Industries Limited (Shin-Etsu Chemical Company Limited)
• Original Assignee: Nissan Chemical Industries Limited (Shin-Etsu Chemical Company Limited)
• Inventors: FURUSHO, Hitoshi
• Granted Patent: US 10,640,772 B2
• Time in Patent Office: Days
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 25/00 Nanomagnetism, e.g. magnetoimpedance, anisotropic magnetoresistance, giant magnetoresistance or tunneling magnetoresistance
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• 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 2310/16 Aptamers
• CPC Class Codes: C12N 2310/3517 Marker; Tag
• CPC Class Codes: G01N 33/50 Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/577 involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens; G01N33/53 - G01N33/576 take precedence

Abstract

The objective of the present invention is to provide a DNA aptamer that can specifically binding to a molecular targeted medicine, a composition comprising the DNA aptamer, and a method for detecting a molecular targeted medicine using the DNA aptamer.

A DNA aptamer comprising specifically binding to a molecular targeted medicine, a composition for detecting a molecular targeted medicine comprising the DNA aptamer, a kid for detecting a molecular targeted medicine, and a method for detecting a molecular targeted medicine comprising using the DNA aptamer.

Claims

1. A DNA aptamer comprising specifically binding to a molecular targeted medicine.

2. A DNA aptamer comprising specifically binding to a molecular targeted medicine, wherein the nucleotide sequence of the DNA aptamer comprises having a sequence comprising 1 to 3 nucleotide substitutions, deletions, or additions.

3. A DNA aptamer specifically binding to a molecular targeted medicine having a nucleotide sequence of:

5′

-P₁-X-P₂-3′

;

wherein X is a nucleotide sequence specifically binding to a molecular targeted medicine or is a sequence comprising 1 to 3 nucleotide substitutions, deletions, or additions, in a nucleotide sequence selected from the sequence, and P₁ and P₂ are a first and a second primer recognition sequence introduced for PCR amplification.

4. The DNA aptamer according to claim 3, wherein P₁ is a first primer recognition sequence shown in SEQ ID NO:

2 and P₂ is a second primer recognition sequence shown in SEQ ID NO;

3.

5. The DNA aptamer according to claim 4 comprising at least one chemical modification selected from the group consisting of a chemical substitution at a sugar chain moiety, chemical substitution at a phosphate ester moiety, and chemical substitution at a nucleic acid base moiety.

6. The DNA aptamer according to claim 5 having a fluorescent labelling at the 5′

end or 3′

end.

7. The DNA aptamer according to claim 6, wherein the fluorescent labelling is one or more kinds selected from green fluorescent protein (GFP), fluorescent protein, fluorescein isothiocyanate (FITC), 6-carboxyfluorescein-aminohexyl, fluorescein derivative, Alexa Fluor 350, Alexa Fluor 405, Alexa Fluor 488, Alexa Fluor 647, Alexa Fluor 750, rhodamine, 6-carboxytetramethylrhodamine, TAMRA (Registered Mark), Phycoerythrin (PE), phycocyanin (PC), PC5, PC7, Cy dyes, Cy3, Cy3.5, Cy5, Cy5.5, Cy7, TexasRed, allophycocyanin (APC), aminomethyl coumarin acetate (AMCA), Marina Blue, Cascade Blue, Cascade Yellow, Pacific Blue, SPRD, Tetramethylrhodamine isothiocyanate (TRITC), R110, mC1B, CellTracker dyes, CFSE, JC-1, PKH, DCFH-DA, DHR, FDA, Calcein AM, nitrobenzoxadiazole (NBD) group, dimethylamino sulphonyl benzooxadiazole group, acridine (Acd), dansyl (Dns), 7-dimethylaminocoumarin-4-acetic acid (DMACA), 5-((2-Aminoethyl)amino)naphthalene-1-sulfonic acid (EDANS), naphthalene, anthracene, and protoporphyrin 9.

8. The DNA aptamer according to claim 6, wherein the fluorescent labelling is a quantum dot.

9. The DNA aptamer according to claim 8 having a fluorescent label at the 5′

or 3′

end, which is adsorbed onto a metal nanoparticle surface.

10. The DNA aptamer according to claim 9, wherein the metal used in the metal nanoparticle is gold, silver, copper, iron, or silicon.

11. The aptamer according to claim 10, wherein the molecular targeted medicine is Bevacizumab and the sequence specifically binding to the molecular targeted medicine is shown in SEQ ID NO:

1.

12. A composition for detecting a molecular targeted medicine comprising the DNA aptamer according to claim 11.

13. A kit for detecting a molecular targeted medicine comprising the DNA aptamer according to claim 11.

14. A method for detecting a molecular targeted medicine comprising using the DNA aptamer according to claim 11.

15. The method for detecting according to claim 14, comprising contacting the DNA aptamer with a sample obtained from a living body selected from a group consisting of blood, blood serum, blood plasma, saliva, and sputum, and detecting the presence of the molecular targeted medicine by observing the response caused by the binding of the sample and the DNA aptamer.

16. The method for detecting according to claim 15, wherein the response is a fluorescent response.

17. The method for detecting according to claim 15, wherein the response is a Raman scattering response.