P16 expression constructs and their application in cancer therapy

• US 6,251,871 B1
• Filed: 08/13/1997
• Issued: 06/26/2001
• Est. Priority Date: 07/17/1995
• Status: Expired due to Fees

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Jin, Xiaomei, Roth, Jack A.
• Primary Examiner(s): Clark, Deborah J.
• Application Number:
  US08/910,722
• Time in Patent Office: 1,413 Days
• Field of Search: 536/23.1, 536/23.5, 435/320.1, 514/44, 424/93.1, 424/93.21
• US Class Current: 514/44.00R
• 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 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C12N 2799/022 where the vector is derived from an adenovirus

Abstract

A variety of genetic constructs are disclosed that will find both in vitro and in vivo use in the area of tumor biology and cancer therapy. In particular, expression constructs are provided that contain a p16 encoding region and other regulatory elements necessary for the expression of a p16 transcript. One version of the expression construct is a replication-deficient adenoviral vector. Also provided are methods for the transformation of cell lines and the inhibition of cancer cell proliferation.

Claims

1. A method of inhibiting tumor growth in a mammal comprising:

(i) providing a pharmaceutical composition comprising (a) an adenoviral vector comprising a promoter functional in eukaryotic cells and a nucleic acid encoding a p16, said nucleic acid being under the control of said promoter and positioned in a sense orientation to said promoter, and (b) a pharmaceutically acceptable buffer, solvent or diluent; and

(ii) administering said pharmaceutical composition to said mammal by direct intratumoral administration.

2. The method according to claim 1, wherein said mammal is a human.

3. The method according to claim 1, wherein said tumor is a lung cancer tumor, a bladder cancer tumor, an esophageal cancer tumor, a pancreatic cancer tumor, a head and neck cancer tumor, a glioma, or a melanoma.

4. The method according to claim 1, wherein said adenoviral vector further comprises a polyadenylation signal 3′

to said p16 encoding nucleic acid.

5. The method according to claim 1, wherein said adenoviral vector is a replication deficient adenovirus.

6. The method according to claim 5, wherein said adenoviral vector lacks at least a portion of the E1 region.

7. The method according to claim 6, wherein said adenoviral vector lacks the entire E1 region.

8. The method according to claim 1, wherein said direct intratumoral administration comprises three doses of adenoviral vector totalling 10¹⁰ PFU.

9. The method according to claim 1, wherein said promoter is selected from the group consisting of CMV IE, SV40 early, RSV LTR, tyrosinase, alpha-fetoprotein, albumin, PSA and PAI-1.

10. The method according to claim 1, wherein said promoter is CMV IE.

11. The method according to claim 1, wherein said nucleic acid encoding p16 is a cDNA.

12. The method according to claim 1, wherein said pharmaceutical composition comprises adenoviral vector at 10⁹ to 10¹¹ PFU/ml.

13. The method according to claim 1, wherein said administration comprises injection.