miRNA for treatment of breast cancer

Patent metadata

Field	Value
Publication number	US-11085044-B2
Application number	US-201615555822-A
Country	US
Kind code	B2
Filing date	Mar 9, 2016
Priority date	Mar 9, 2015
Publication date	Aug 10, 2021
Grant date	Aug 10, 2021

Abstract

The presently-disclosed subject matter relates to RNA-based composition and method to treat breast cancer in a subject. More particularly, the presently disclosed subject matter relates to a RNA nanostructure and composition containing a multiple branched RNA nanoparticle, a breast cancer targeting module, and an effective amount of a breast cancer therapeutic agent. Further, the presently disclosed subject matter relates to a method of using the RNA nanoparticle composition to treat breast cancer in a subject having or at risk of having breast cancer.

First claim

What is claimed is:

1. An artificial RNA nanostructure molecule, comprising: a multiple branched RNA junction motif comprising at least one RNA oligonucleotides, and a breast cancer targeting module coupled to the RNA junction motif, 5′-UUG CCA UGU GUA UGU GGG AUC CCG CGG CCA UGG CGG CCG GGA G-3′ (SEQ ID NO: 5).

2. The molecule of claim 1 , further comprising at least one bioactive agent coupled to the RNA junction motif.

3. The molecule of claim 1 , wherein the bioactive agent is a therapeutic agent.

4. The molecule of claim 1 , wherein the RNA oligonucleotide comprises at least one chemical modification at the 2′ position.

5. The molecule of claim 1 , wherein the modification comprises 2′Fluoro, 2′Amine, or 2′O-Methyl.

6. The molecule of claim 1 , wherein the multiple branched RNA junction motif is a three-branched RNA junction motif.

7. The molecule of claim 1 , wherein the multiple branched RNA further comprises sequence 5′-CCC ACA UAC UUU GUU GAU CCG CCU UAG UAA CGU GCU UUG AUG UCG AUU CGA CAG GAG GC-3′(SEQ ID NO: 6); 5′-GATAAGCT CTC CCG GCC GCC ATG GCC GCG GGA T-3′ (SEQ ID NO: 7); 5′-CTC CCG GCC GCC ATG GCC GCG GGA T-3′ (SEQ ID NO: 8); 5′-AUC CCG CGG CCA UGG CGG CCG GGA G-3′ (SEQ ID NO: 9); or a combination thereof.

8. The molecule of claim 1 , wherein the diameter of the molecule is at least about 40 nm or less.

9. The molecule of claim 1 , wherein the molecule has zeta potential ranging from about −100 mV to about 100 mV.

10. The molecule of claim 6 , wherein a branch of the three-branched RNA junction motif comprises an a 3WJ RNA module (SEQ ID NO: 1); a b3WJ RNA module (SEQ ID NO: 2); or a c3WJ RNA module (SEQ ID NO: 3).

11. The molecule of claim 6 , wherein the three-branched RNA junction motif comprises an a 3WJ RNA module (SEQ ID NO: 1); a b3WJ RNA module (SEQ ID NO: 2); and a c3WJ RNA module (SEQ ID NO: 3).

12. The molecule of claim 1 , wherein the breast cancer targeting module comprises a ligand that binds to at least one breast cancer cell surface marker.

13. The molecule of claim 12 , wherein the ligand binds to a folate receptor, an epidermal growth factor receptor 2 (ErbB-2/HER2), an epidermal growth factor receptor (EGFR), a HER2, or a combination thereof.

14. The molecule of claim 12 , wherein the ligand is an aptamer.

15. The molecule of claim 14 , wherein the aptamer binds to EGFR, PDGFR, folate receptor, or a combination thereof.

16. The molecule of claim 14 , wherein the ligand is a EGFR targeting aptamer.

17. The molecule of claim 1 , wherein the ligand has sequence 5′-G CCU UAG UAA CGU GCU UUG AUG UCG AUU CGA CAG GAG GC-3′(SEQ ID NO: 10).

18. The molecule of claim 1 , wherein the targeting module is a folate.

19. The molecule of claim 18 , wherein the folate is folic acid, 5-methyltetrahydro folate, 5-formyltetrahydrofolate, dihydrofolate, tetrahydrofolate, or a combination thereof.

20. The molecule of claim 2 , wherein the bioactive agent is a drug, a fluorescent dye, or a chemical, or a combination thereof.

21. The molecule of claim 2 , wherein the bioactive agent is a siRNA, a miRNA, an anti-miRNA, a ribozyme RNAs, or an antisense RNA.

22. The molecule of claim 2 , wherein the bioactive agent is directed to a breast cancer marker.

23. The molecule of claim 21 , wherein the bioactive agent is a siRNA sequence or a microRNA sequence.

24. The molecule of claim 21 , wherein the bioactive agent is a miRNA molecule for a miRNA comprising miR-9, miR-10b, miR-21, miR-17, miR-26, let-7a, miR-25, miR-34a, miR-124, miR-145, or miR-181b.

25. The molecule of claim 21 , wherein the microRNA sequence is an anti-miR-21 sequence.

26. The molecule of claim 25 , wherein the anti-miR-21 sequence comprises 5′-GATAAGCT-3′ (SEQ ID NO: 11).

27. The molecule of claim 25 , wherein the anti-miRNA comprises an anti-miRNA locked nucleic acid (LNA) molecule.

28. The molecule of claim 21 , wherein the anti-miRNA LNA molecule comprises sequence 5′-GATAAGCT-3′, 5′-AGCACTTT-3′, or 5′-ATTTGCAC-3′.

29. The molecule of claim 23 , wherein the siRNa binds to a mRNA molecule that encodes RAS, cMET, HER2, MDM2, PIK3CA, AKT, CDK4, or a combination thereof.

30. The molecule of claim 1 , wherein the RNA nanostructure inhibit breast cancer cells proliferation.

31. A nucleic acid composition, comprising a therapeutically effective amount of the RNA nanostructure of claim 1 and a pharmaceutically acceptable carrier.

32. A nanoparticle delivery system, comprising a RNA nanostructure of claim 1 and a pharmaceutically acceptable carrier.

33. A method of treating a breast tumor in a subject having or at risk of developing a breast cancer, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a molecule of claim 1 and a pharmaceutically acceptable carrier.

34. The method of claim 33 , wherein the breast cancer is triple negative breast cancer.

Assignees

• Univ Kentucky Res Found

Inventors

• Guo Peixuan
• Shu Dan
• Shu Yi
• Li Hui
• Haque Farzin

Classifications

• C07H21/02 Primary

  with ribosyl as saccharide radical · CPC title

• C12N2310/52

  branched · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• C12N2320/30

  Special therapeutic applications · CPC title