Targeting of human glucocorticoid receptor beta in cancer

• US 10,457,947 B2
• Filed: 03/07/2017
• Issued: 10/29/2019
• Est. Priority Date: 03/10/2016
• Status: Active Grant

• Current Assignee: University of Toledo
• Original Assignee: The University of Toldeo
• Inventors: Hinds, Jr., Terry D., McBeth, Lucien
• Primary Examiner(s): Vivlemore, Tracy
• Application Number:
  US16/082,188
• Publication Number:
  US 20190085337A1
• Time in Patent Office: 966 Days
• Field of Search: None
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/162 from virus
• CPC Class Codes: A61K 47/6455 Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/163 Regulatory proteins, e.g. tat, nef, rev, vif, vpu, vpr, vpt, vpx
• CPC Class Codes: C07K 14/721 Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor
• CPC Class Codes: C12N 15/113 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 Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/113 targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/3181 Peptide nucleic acid, PNA
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12Q 1/68 involving nucleic acids

Abstract

A peptide nucleic acid (PNA) referred to herein as Sweet-P, compositions comprising the methods of making the same, mid methods of using the same, are described.

Claims

1. A composition comprising a peptide nucleic acid (PNA) consisting of the sequence TGCCATACACAGTAT [SEQ ID NO:

1].

2. The composition of claim 1, wherein the PNA is attached to a solubility enhancing molecule.

3. The composition of claim 2, wherein the solubility enhancing molecule comprises an O-linker.

4. The composition of claim 1, wherein the PNA is conjugated to a cell-penetrating peptide.

5. The composition of claim 4, wherein the cell-penetrating peptide comprises a modified tat protein.

6. The composition of claim 5, wherein the modified tat protein consists of the amino acid sequence VQRKRQKLMP [SEQ ID NO:

2].

7. The composition of claim 1, wherein the PNA is attached to an 0-linker and to a modified tat protein consisting of the amino acid sequence VQRKRQKLMP [SEQ ID NO:

2].

9. A pharmaceutical composition comprising:

an effective amount of the composition of claim 1; and

a pharmaceutically acceptable carrier, diluent, or adjuvant.

13. A method of treating a GRβ

-related disease, the method comprising;

administering an effective amount of an agent comprising the composition of claim 1 to a subject in need thereof, wherein the effective amount of the agent binds to the glucorticoid receptor gene and blocks miR144 binding to GRβ

in the subject, and treating a GRβ

-related disease in the subject.

14. The method of claim 13, wherein the agent is conjugated to a modified tat protein.

15. The method of claim 13, wherein the GRβ

-related disease is selected from the group consisting of;

bladder cancer, prostate cancer, lung cancer, liver cancer, fatty liver disease, glioblastoma, leukemia, lupus, and asthma.

16. A method of hindering migration of a GRβ

-related disease, the method comprising administering an effective amount of a composition of claim 1 to a subject in need thereof and hindering migration of a GRβ

-related disease.

17. The method of claim 16, wherein the GRβ

-related disease is selected from the group consisting of bladder cancer, prostate cancer, lung cancer, liver cancer, fatty liver disease, glioblastoma, and leukemia.

18. The method of claim 16, wherein the subject is a human subject.

19. A method for regulating GRβ

expression in cells, the method comprising administering an effective amount of a composition of claim 1 to cells and regulating expression of GRβ

in the cells.

20. The method of claim 19, wherein GRβ

is suppressed in the cells.

21. The method of claim 19, wherein the cells are cancer cells or liver cells.

22. The method of claim 21, wherein the cancer cells are selected from the group consisting of bladder cancer, prostate cancer, lung cancer, liver cancer, glioblastoma, and leukemia.

23. The method of claim 19, wherein the cells are in a human subject.

8. A peptide nucleic acid (PNA) consisting of the sequence TGCCATACACAGTAT [SEQ ID NO:

1].

10. A method of modulating GRβ

expression in cells, the method comprising;

administering to cells an effective amount of an agent which binds to the glucocorticoid receptor gene and is capable of blocking miR144 binding to GRβ

, and modulating GRβ

expression in the cells;

wherein the agent comprises a PNA consisting of the sequence TGCCATACACAGTAT [SEQ ID NO;

1].

11. The method of claim 10, wherein the cells are cancer cells.

12. The method of claim 10, wherein the cells are in a human subject.

24. A kit for preparing a pharmaceutical composition, the kit comprising:

a first container housing a peptide nucleic acid (PNA) consisting of the sequence TGCCATACACAGTAT [SEQ ID NO;

1]; and

a second container housing an O-linker or a modified tat protein consisting of the amino acid sequence VQRKRQKLMP [SEQ ID NO;

2].