Method of treating hyperproliferation of cells using a nucleolin specific aptamer

20140213636

14/035003

September 24, 2013

Improved G-rich oligonucleotide (GRO) aptamers specific to nucleolin, a method of preparing the aptamers, and a use of the aptamers for diagnosing and/or treating a nucleolin-associated disease, are provided.

POSTECH ACADEMY-INDUSTRY FOUNDATION (POHANG-SI, KR)

1. A method of treating a cancer, comprising administering a nucleolin-specific aptamer a subject in need of treating a cancer, wherein the nucleolin-specific aptamer has the nucleotide sequence of SEQ ID NO: 3, and one or more thymidines(T) present in the nucleotide sequence are independently substituted with a modified pyrimidine nucleoside, and wherein the modified pyrimidine nucleoside is deoxyuridine(dU), deoxycytidine(dC), uridine(U), or cytidine(C) having a hydrophobic group at 5-position: [table included] wherein each N is absent or 1 to 20 nucleosides, which is independently selected from the group consisting of adenosine(A), thymidine(T)/uridine(U), cytidine(C), and guanosine(G).

2. The method according to claim 1, the nucleolin-specific aptamer has the nucleotide sequence of SEQ ID NO: 1 or 2, and one or more thymidines(T) present in the nucleotide sequence are independently substituted with a modified pyrimidine nucleoside, and wherein the modified pyrimidine nucleoside is deoxyuridine(dU), deoxycytidine(dC), uridine(U), or cytidine(C) having a hydrophobic group at 5-position.

3. The method according to claim 1, wherein the hydrophobic group is selected from the group consisting of a benzyl group, a naphthyl group, and a pyrrolebenzyl group.

4. The method according to claim 1, wherein the modified pyrimidine nucleoside is selected from the group consisting of 5-(N-benzylcarboxyamide)-2′-deoxyuridine (called BzdU), 5-(N-naphthylcarboxyamide)-2′-deoxyuridine (called NapdU), 5-(N-4-pyrrolebenzylcarboxyamide)-2′-deoxyuridine (called 4-PBdU), 5-(N-benzylcarboxyamide)-2′-deoxycytidine (called BzdC), 5-(N-naphthylcarboxyamide)-2′-deoxycytidine (called NapdC), 5-(N-4-pyrrolebenzylcarboxyamide)-2′-deoxycytidine (called 4-PBdC), 5-(N-benzylcarboxyamide)-2′-uridine (called BzU), 5-(N-naphthylcarboxyamide)-2′-uridine (called NapU), 5-(N-4-pyrrolebenzylcarboxyamide)-2′-uridine (called 4-PBU), 5-(N-benzylcarboxyamide)-2′-cytidine (called BzC), 5-(N-naphthylcarboxyamide)-2′-cytidine (called NapC), and 5-(N-4-pyrrolebenzylcarboxyamide)-2′-cytidine (called 4-PBC).

5. The method according to claim 1, wherein the nucleolin-specific aptamer has SEQ ID NO: 3, and at least two thymidines present in 9th to 18th positions of SEQ ID NO: 3 are substituted with the modified pyrimidine nucleoside, and the positions are counted starting from ‘G’ after ‘N’ at 5′-end.

6. The method according to claim 1, wherein the nucleolin-specific aptamer has SEQ ID NO: 1 or SEQ ID NO: 2, and at least two thymidines present in 12th to 18th positions of SEQ ID NO: 1 or 9th to 18th positions of SEQ ID NO: 2 are substituted with the modified pyrimidine nucleoside.

7. The method according to claim 1, wherein the nucleolin-associated cancer is selected from the group consisting of leukemias, lymphomas, myeloproliferative disorders, carcinomas of solid tissue, sarcomas, melanomas, adenomas, hypoxic tumors, squamous cell carcinomas of the mouth, throat, larynx, or lung, genitourinary cancers, hematopoietic cancers, head and neck cancers, and nervous system cancers, and benign lesions.

8. A method of inhibiting nucleolin, comprising administering a nucleolin-specific aptamer to a subject in need of inhibiting nucleolin or a sample comprising nucleolin-expressing cells, wherein the nucleolin-specific aptamer has the nucleotide sequence of SEQ ID NO: 3, and one or more thymidines(T) present in the nucleotide sequence are independently substituted with a modified pyrimidine nucleoside, and wherein the modified pyrimidine nucleoside is deoxyuridine(dU), deoxycytidine(dC), uridine(U), or cytidine(C) having a hydrophobic group at 5-position: [table included] wherein each N is absent or 1 to 20 nucleosides, which is independently selected from the group consisting of adenosine(A), thymidine(T)/uridine(U), cytidine(C), and guanosine(G).

9. A method of inhibiting hyperproliferation of cell cased by nucleolin, comprising administering a nucleolin-specific aptamer to a subject in need of inhibiting hyperproliferation of cell cased by nucleolin or a sample comprising nucleolin-expressing cells, wherein the nucleolin-specific aptamer has the nucleotide sequence of SEQ ID NO: 3, and one or more thymidines(T) present in the nucleotide sequence are independently substituted with a modified pyrimidine nucleoside, and wherein the modified pyrimidine nucleoside is deoxyuridine(dU), deoxycytidine(dC), uridine(U), or cytidine(C) having a hydrophobic group at 5-position: [table included] wherein each N is absent or 1 to 20 nucleosides, which is independently selected from the group consisting of adenosine(A), thymidine(T)/uridine(U), cytidine(C), and guanosine(G).

514/44/R

12

edspap.20140213636