Southern Research Institute

The Uab Research Foundation

University of Alabama

Other

Cornell Research Foundation Inc.

Cornell Center for Technology, Enterprise and Commercialization

Southern Research Inst.

William B Parker

Eric J Sorscher

Leonard L Bennett, Jr

John A Secrist, Iii

Steven Ealick

Vijayakrishna K Gadi

William Waud

John A Montgomery

David I Scheer

L Lee Bennett

Y Fumer Shealy

The use of a purine nucleoside phosphorylase or nucleoside hydrolase or a vector encoding expression of one of these enzymes is detailed along with the use of a prodrug cleaved by the purine nucleoside phosphorylase or nucleoside hydrolase for the preparation of a direct injection inhibition of replicating or non-replicating targeted cells. The targeted cells do not normally express the introduced purine nucleoside phosphorylase or nucleoside hydrolase. The enzyme and prodrug are amenable to intermixing and injection as a single dose or as separate injection or administration to the targeted cells. The substance and prodrug efficacy are enhanced through exposure of the targeted cells to X-ray radiation. Administration of a prodrug regardless of administration route to the targeted cells is effective in combination with X-ray radiation therapy to kill or inhibit function of the targeted cells.

Enhanced therapeutic usage of a purine nucleoside phosphorylase or nuceloside hydrolase prodrug

Enhanced therapeutic usage of a purine nucleoside phosphorylase or nucleoside hydrolase prodrug

A process for inhibiting a mammalian cancerous cell or virally infected cell includes providing apurine nucleoside phosphorylase enzyme or a tail mutant purine nucleoside phosphorylase enzyme in proximity to the mammalian cancerous cell or the virally infected cell and exposing the enzyme to a purine nucleoside phosphorylase enzyme cleavable substrate to yield a cytotoxic purine analog. The process includes introducing to the cell a vector containing the phosphorylase enzyme, or a DNA sequence coding for the same and delivering to the cell an effective amount of the substrate such as 9-(&#x3b2;-D-arabinofuranosyl)-2-fluoroadenine (F-araA).

Purine nucleoside phosphorylase as enzymatic activator of nucleoside prodrugs

A host cell stably transformed or transfected by a vector including a DNA sequence encoding for mutant purine nucleoside cleavage enzymes is provided. The transformed or transfected host cell can be used in combination with a purine substrate to treat tumor cells and/or virally infected cells. A nucleotide sequence encoding mutantderived purine nucleoside phosphorylase proteins which can be used in conjunction with an appropriate substrate to produce toxins which impair abnormal cell growth is also provided. A method is detailed for the delivery of toxin by generation within target cells or by administration and delivery to the cells from without. Novel purine nucleosides are detailed that yield a cytotoxic purine upon enzymatic cleavage. A synthetic process for nucleosides is also detailed.

Mutant purine nucleoside phosphorylase proteins and cellular delivery thereof

The present invention provides a procaryotic host cell stably transformed or transfected by a vector including a DNA sequence encoding for mutant purine nucleoside phosphorylase or hydrolase. The transformed or transfected procaryotic host cell can be used in combination with a purine substrate to treat tumor cells and/or virally infected cells. The present invention provides nucleotide sequences encoding mutantderived purine nucleoside phosphorylase proteins which can be used in conjunction with an appropriate substrate to produce toxins which impair abnormal cell growth. The invention provides for delivery of the toxin by generation within target cells or by administration and delivery to the cells from without.

The present invention provides a procaryotic host cell stably transformed or transfected by a vector including a DNA sequence encoding for purine nucleoside phosphorylase or hydrolase. The transformed or transfected procaryotic host cell can be used in combination with a purine substrate to treat tumor cells and/or virally infected cells.

Recombinant bacterial cells for delivery of PNP to tumor cells

The present invention provides a method of killing replicating or non-replicating, transfected or transduced mammalian cells and bystander cells, comprising: (a) transfecting or transducing mammalian cells with a nucleic acid encoding a non-human purine cleavage enzyme; and (b) contacting the transfected or transduced cells with an effective amount of a substrate for the purine cleavage enzyme, wherein the substrate is non-toxic to mammalian cells and is cleaved by the enzyme to yield a purine toxic to the targeted mammalian cells and bystander cells, to kill the mammalian cells expressing the enzyme and the bystander cells. Further provided is a vector comprising a DNA sequence coding for a non-human purine nucleoside phosphorylase protein and the vector is capable of replication and/or expression in a host which comprises, in operable linkage: a) optionally, an origin of replication; b) a promoter; and c) a DNA sequence coding for said protein.

Purine nucleoside phosphorylase gene therapy for human malignancy

Method for prophylaxis and treatment of a viral infection characterized by the administration of a composition comprising a substantial molar excess of the D-stereoisomer of 2'CdG over the L-stereoisomer.

Methods of treatment of viral infections using carbocyclic

The invention provides a method of killing replicating or non-replicating, transfected or transduced mammalian cells and bystander cells, comprising the following steps: (a) transfecting or transducing mammalian cells with a nucleic acid encoding a non-human purine nucleoside phosphorylase (PNP); and (b) contacting the transfected or transduced cells with an amount of a substrate for the purine nucleoside phosphorylase sufficient to produce a toxic purine base-analog thereby killing the transfected or transduced cells and bystander cells. In the present method of killing cells, the non-human purine nucleoside phosphorylase can be an E. coli purine nucleoside phosphorylase. The method of the invention described above can utilize a substrate that is a purine nucleoside analog. For instance, in the method provided in the Examples, the substrate is 9-(.beta.-D-2-deoxyerythropentofuranosyl)-6-methylpurine (MeP-dR).

Growing community of inventors

Birmingham, AL, United States of America

William B Parker