Biosynthetic oncolytic molecules and uses therefor

• US 20050048614A1
• Filed: 07/28/2004
• Published: 03/03/2005
• Est. Priority Date: 06/13/2000
• Status: Abandoned Application

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation
• Inventors: Ashkar, Samy, Hikita, Sherry, Dehni, Ghassan
• Application Number:
  US10/900,512
• Publication Number:
  US 20050048614A1
• Time in Patent Office: Days
• US Class Current: 435/69.100
• 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: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus

Abstract

Biosynthetic oncolytic molecules including an apoptotic domain derived from osteopontin have been developed which are capable of promoting cellular apoptosis.

Claims

1. A biosynthetic oncolytic molecule comprising an apoptotic component derived from osteopontin and at least one biomodular component, forming a molecule which promotes apoptosis, comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:

8, 10, 12, 15, 16, 17, 18, and conservative substitutions therein.

2. The oncolytic molecule of claim 3, wherein the apoptotic component comprises an amino acid sequence up to 50 amino acid residues.

3. The oncolytic molecule of claim 1, wherein the biomodular component is selected from the group consisting of a signal peptide, a linker domain, a golgi processing domain, a heparin binding domain, and a collagen binding domain.

4. The oncolytic molecule of claim 1, comprising an apoptotic component, a first biomodular component and a second biomodular component.

5. The oncolytic molecule of claim 4, wherein the first and second biomodular components are selected from the group consisting of a signal peptide, a linker domain and a golgi processing domain.

6. The oncolytic molecule of claim 4, further comprising a third biomodular component.

7. The oncolytic molecule of claim 6, wherein the third biomodular component is a heparin binding domain or a collagen binding domain.

8. The oncolytic molecule of claim 1, wherein the molecule modulates an apoptotic response selected from the group consisting of modulation of chromatin structure, cell viability, and cell lysis.

9. The oncolytic molecule of claim 1, wherein the molecule enhances an apoptotic response.

10. The oncolytic molecule of claim 9, wherein the apoptotic response is cell lysis.

11. The oncolytic molecule of claim 1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:

8, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, and SEQ ID NO;

18.

14. An expression vector comprising the nucleic acid molecule of claim 11.

17. A method of producing an oncolytic molecule, comprising culturing the host cell of claim 14 under conditions such that the oncolytic molecule is produced.

18. The method of claim 17, further comprising isolating the oncolytic molecule from the medium or the host cell.

12. An isolated nucleic acid molecule encoding a biosynthetic oncolytic molecule comprising an apoptotic component derived from osteopontin and at least one biomodular component, forming a molecule which promotes apoptosis, comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:

8, 10, 12, 15, 16, 17, 18, and conservative substitutions therein.

13. The molecule of claim 12 comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOS:

7, 9, 11, 13 and 17.

15. An expression vector comprising the nucleic acid molecule selected from the group consisting of SEQ ID NO:

7, SEQ ID NO;

9, SEQ ID NO;

11, SEQ ID NO;

13, and SEQ ID NO;

17.

16. A host cell comprising the vector of claim 15.

19. A method of modulating an apoptotic response in a cell comprising contacting the cell with a biosynthetic oncolytic molecule comprising an apoptotic component derived from osteopontin and at least one biomodular component, forming a molecule which promotes apoptosis, comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:

8, 10, 12, 15, 16, 17, 18, and conservative substitutions therein in an amount and for a period of time to induce an apoptotic response.