Cancer therapies and diagnostics

Patent metadata

Field	Value
Publication number	US-10822374-B2
Application number	US-201113884709-A
Country	US
Kind code	B2
Filing date	Nov 14, 2011
Priority date	Nov 12, 2010
Publication date	Nov 3, 2020
Grant date	Nov 3, 2020

Abstract

Compositions and methods for increasing p53-dependent transcriptional activity in a cell.

First claim

What is claimed is:

1. A method for treating a cancer expressing both HDM2 and HDMX in a human subject in need thereof, the method comprising:
administering to the human subject a HDMX modulating agent and a HDM2 modulating agent,
wherein the HDMX modulating agent comprises an internally cross-linked polypeptide comprising at least nine contiguous amino acids of the amino acid sequence Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Phe 6 -Xaa 7 -Xaa 8 -Xaa 9 -Trp 10 -Xaa 11 -Xaa 12 -Leu 13 -Xaa 14 -Xaa 15 -Xaa 16 (SEQ ID NO: 22), wherein the internally cross-linked polypeptide includes Phe 6 , Trp 10 , and Leu 13 , and wherein:
Xaa 1 is Leu or Gln, or is missing;
Xaa 2 is Ser or is missing;
Xaa 3 is Gln or is missing;
Xaa 4 is Glu, Gln, or a hydrophobic amino acid;
Xaa 5 is Thr or Ala;
Phe 6 is Phe;
Xaa 7 is Ser or a stapling amino acid;
Xaa 8 is Asp or Asn;
Xaa 9 is Leu or a hydrophobic amino acid;
Trp 10 is Trp;
Xaa 11 is Lys, a positively charged amino acid, or a hydrophobic amino acid;
Xaa 12 is Leu, Lys, or Ala;
Leu 13 is Leu;
Xaa 14 is Pro or a stapling amino acid;
Xaa 15 is Glu, Gln, or Ala;
Xaa 16 is Asn or is missing;
wherein the side chains of at least two amino acids of the amino acid sequence separated by two, three, or six amino acids are replaced by an internal cross-link, and
wherein the internally cross-linked polypeptide binds HDMX, is alpha helical, is neutral to positively charged, is cell permeable, and is not ubiquitinylated; and
wherein the HDM2 modulating agent is Nutlin, a spiro-oxindole, a benzodiazepinedione, a terphenyl-based helical mimetic, a miniature protein, or a HDM2 targeting microRNA.

2. The method of claim 1 , wherein the HDMX modulating agent is an internally cross-linked polypeptide comprising at least nine contiguous amino acids of the amino acid sequence Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Phe 6 -Xaa 7 -Xaa 8 -Xaa 9 -Trp 10 -Xaa 11 -Xaa 12 -Leu 13 -Xaa 14 -Xaa 15 -Xaa 16 (SEQ ID NO: 22), wherein the internally cross-linked polypeptide includes Phe 6 , Trp 10 , and Leu 13 , and wherein:
Xaa 1 is Gln;
Xaa 2 is Ser;
Xaa 3 is Gln;
Xaa 4 is Gln;
Xaa 5 is Thr;
Phe 6 is Phe;
Xaa 7 is a stapling amino acid;
Xaa 8 is Asn;
Xaa 9 is Leu;
Trp 10 is Trp;
Xaa 11 is Arg;
Xaa 12 is Leu;
Leu 13 is Leu;
Xaa 14 is a stapling amino acid;
Xaa 15 is Gln; and
Xaa 16 is Asn.

3. The method of claim 1 , wherein the HDMX modulating agent comprises the amino acid sequence of SEQ ID NO: 10, except having 1, 2, or 3 amino acid substitutions.

4. The method of claim 1 , further comprising administering the HDMX modulating agent and the HDM2 modulating agent with a pharmaceutically acceptable carrier, adjuvant, or vehicle.

5. The method of claim 1 , further comprising administering the HDMX modulating agent and the HDM2 modulating agent with an additional therapeutic agent, in amounts effective for achieving a modulation of disease or disease symptoms.

6. The method of claim 1 , wherein the HDMX modulating agent comprises the amino acid sequence of SEQ ID NO: 10.

7. The method of claim 1 , wherein the HDMX modulating agent consists of the amino acid sequence of SEQ ID NO: 10.

8. A method for increasing p53 activity in a cell expressing both HDM2 and HDMX and in which p53 activity is not substantially increased upon treatment with a HDM2 modulating agent, the method comprising:
contacting the cell with a HDMX modulating agent and a HDM2 modulating agent, thereby increasing p53 activity in the cell,
wherein the HDMX modulating agent is an internally cross-linked polypeptide comprising the amino acid sequence of SEQ ID NO: 10, except having 0-13 amino acid substitutions, wherein positions 6, 10, and 13 of SEQ ID NO: 10 are not substituted and positions 7 and 14 of SEQ ID NO: 10 are stapling amino acids, and wherein the internally cross-linked polypeptide binds HDMX, is alpha helical, is neutral to positively charged, is cell permeable, and is not ubiquitinylated, and
wherein the HDM2 modulating agent is Nutlin, a spiro-oxindole, a benzodiazepinedione, a terphenyl-based helical mimetic, a miniature protein, or a HDM2 targeting microRNA.

9. A method of treating a cancer expressing functional p53 and also expressing both HDM2 and HDMX in a human subject in need thereof, the method comprising administering to the human subject a HDMX modulating agent and a HDM2 modulating agent, wherein the HDMX modulating agent is an internally cross-linked polypeptide comprising the amino acid sequence of SEQ ID NO: 10, except having 0-13 amino acid substitutions, wherein positions 6, 10, and 13 of SEQ ID NO: 10 are not substituted and positions 7 and 14 of SEQ ID NO: 10 are stapling amino acids, and wherein the internally cross-linked polypeptide binds HDMX, is alpha helical, is neutral to positively charged, is cell permeable, and is not ubiquitinylated, and
wherein the HDM2 modulating agent is Nutlin, a spiro-oxindole, a benzodiazepinedione, a terphenyl-based helical mimetic, a miniature protein, or a HDM2 targeting microRNA.

10. The method of claim 9 , further comprising administering to the human subject an additional therapeutic agent, in amounts effective for achieving a modulation of disease or disease symptoms.

11. The method of claim 1 , wherein the HDM2 modulating agent is Nutlin 3.

12. The method of claim 1 , wherein the HDM2 modulating agent is a spiro-oxindole, benzodiazepinedione, or a terphenyl-based helical mimetic.

13. The method of claim 11 , wherein the Nutlin-3 is Nutlin-3a.

14. The method of claim 1 , wherein the miniature protein is p53AD 15-31 .

15. The method of claim 1 , wherein the HDM2 modulating agent is a HDM2 targeting microRNA.

16. The method of claim 15 , wherein the HDM2 targeting microRNA is miRNA miR-192.

17. The method of claim 8 , wherein the HDM2 modulating agent is Nutlin 3.

18. The method of claim 9 , wherein the internally cross-linked polypeptide comprises the amino acid sequence of SEQ ID NO: 10 except having 0-10 amino acid substitutions.

19. The method of claim 9 , wherein the internally cross-linked polypeptide comprises the amino acid sequence of SEQ ID NO: 10 except having 0-6 amino acid substitutions.

20. The method of claim 9 , wherein the internally cross-linked polypeptide comprises the amino acid sequence of SEQ ID NO: 10 except having 0-3 amino acid substitutions.

21. The method of claim 9 , wherein the cancer is a fibrosarcoma, myosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, gastric cancer, esophageal cancer, rectal cancer, pancreatic cancer, ovarian cancer, prostate cancer, uterine cancer, cancer of the head and neck, skin cancer, brain cancer, squamous cell carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, testicular cancer, small cell lung carcinoma, non-small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, leukemia, lymphoma, or Kaposi sarcoma.

22. The met

Assignees

• Walensky Loren D
• Bernal Federico
• Wahl Geoffrey
• Wade Mark
• Dana Farber Cancer Inst Inc
• Salk Inst For Biological Studi

Inventors

• Walensky Loren D
• Bernal Federico
• Wahl Geoffrey
• Wade Mark

Classifications

• A61K38/1709 Primary

  from mammals · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• C07K4/12 Primary

  from animals; from humans · CPC title