Robert Allen Malone

US Patent:

20040009947, Jan 15, 2004

Filed:

May 27, 2003

Appl. No.:

10/445034

Inventors:

Robert Malone - Rockville MD, US
Jill Malone - Rockville MD, US

International Classification:

A61K048/00
C12P021/02
C12N015/88
C12N015/85
C12N005/08

US Classification:

514/044000, 435/455000, 435/366000, 435/458000

Abstract:

The discovery of simple, nontoxic, and pharmaceutically defined methods for genetic modification of cells and tissues would enable development of a variety of molecular medicines. “Free”, ‘direct’, or ‘naked’ polynucleotide administration is a simple, apparently safe, and pharmaceutically defined polynucleotide delivery method. Murine, macaque, and clinical human experiments have demonstrated transfection of various tissues, such as respiratory tissues, after direct application of ‘free’ polynucleotide. However, direct DNA transfection is relatively inefficient in comparison to many transduction systems. The invention herein is directed to transfection enhancing agents which augment the transfection activity of ‘free’ polynucleotide, thereby facilitating the development of simple and safe alternatives to tissue transfection, more particularly respiratory tissue transfection. The experiments described herein indicate that nucleases, both extra- and intra-cellular, present in many biological fluids, such as respiratory fluid, accelerate clearance of biologically active plasmid from the tissue, and that co-administration of a nuclease inhibitor together with free polynucleotide results in marked enhancement of expression of the polynucleotide of interest. These findings support the disclosed invention of an improved polynucleotide delivery system, a ‘free’ plasmid-based transfection technology.

US Patent:

61108987, Aug 29, 2000

Filed:

May 23, 1997

Appl. No.:

8/862632

Inventors:

Robert W. Malone - Baltimore MD
Jill G. Malone - Baltimore MD

Assignee:

University of Maryland, Baltimore - Baltimore MD

International Classification:

A61K 4800
C17N 1500
C12Q 170

US Classification:

514 44

Abstract:

The invention consists of a method for inducing production of a mucosal immune response in a host by administration of an antigen-encoding polynucleotide preparation, comprising DNA or RNA encoding an antigenic epitope to a mucosal inductor site in the mucosal tissue of the host. Naked DNA may be administered directly to mucosa, for instance in saline drops, or in a recombinant gene expression vector. Preferably, the recombinant gene expression vectors are not capable of replication or dessimination. The invention also includes the use of live viral vaccines wherein the viruses include immunostimulatory polynucleotides of the invention. According to a preferred method of the invention, a target protein antigen is administered through its expression by a recombinant gene expression vector.