Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Carbohydrate doai
Patent
1997-01-31
2000-02-15
Campell, Bruce R.
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Carbohydrate doai
435 691, 435 697, 435 698, 4353201, 435325, 435455, 536 232, 536 234, 536 237, 536 241, 424 941, 424 9463, A01N 4304
Patent
active
060253401
DESCRIPTION:
BRIEF SUMMARY
INTRODUCTION AND BACKGROUND TO THE INVENTION
The present invention relates to gene directed enzyme prodrug therapy (GDEPT) and its use in the treatment of disease, including tumours.
A therapeutic approach termed "virus-directed enzyme prodrug therapy" (VDEPT) has been proposed as a method for treating tumour cells in patients using prodrugs. Tumour cells are targeted with a viral vector carrying a gene encoding an enzyme capable of activating a prodrug. The gene may be transcriptionally regulated by tissue specific promoter or enhancer sequences. The viral vector enters tumour cells and expresses the enzyme, in order that a prodrug is converted to an active drug within the tumour cells (Huber et al, Proc. Natl. Acad. Sci. USA (1991) 88, 8039). Alternatively, non-viral methods for the delivery of genes have been used. Such methods include calcium phosphate co-precipitation, microinjection, liposomes, direct DNA uptake, and receptor-mediated DNA transfer. These are reviewed in Morgan & French Anderson, Annu. Rev. Biochem., 1993,62;191. The term "GDEPT" (gene-directed enzyme prodrug therapy) is used to include both viral and non-viral delivery systems.
The success of a GDEPT system relies upon two limiting factors. The system requires cells which need to be targeted by the vector to be infected. If a cell does not become infected then no active drug will be produced within it so that in order to be killed active drug will need to enter it by a bystander effect, having been produced in another infected cell. Not all active drugs when produced inside a cell will be capable of leaving that cell in order to do this. Further, it is a requirement that the prodrug enters the cell. Some prodrugs may not be capable of crossing the membrane of a cell.
DISCLOSURE OF THE INVENTION
In order to overcome these problems, the present invention provides a two component system for use in association with one another comprising:
The vector may be a RNA or DNA vector. It may be derived from a viral vector, including any suitable vector available in the art for targeting tumour cells.
The invention also provides the system of the invention for use in a method of treatment of a patient, and a method of treating a tumour in a patient in need of treatment which comprises administering to said patient an effective amount of a vector encoding an enzyme capable of being expressed on the surface of a cell and a prodrug capable of being converted by said enzyme to an active drug.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1. Expression of CPG2(surface) in NIH3T3 cells NIH3T3 cells were transfected with MLV.beta.plink (lane 1) or with MLVCPG2(surface) (lane 2). The cells were extracted into buffer A and samples of each were electrophoresed in 10% w/v polyacrylamide gels, transferred to nitrocellulose and probed with a CPG2 specific polyclonal antibody. The position of migration of CPG2(surface) is shown and the positions of migration of standard molecular mass markers (in kDa) are shown to the left of the figure
FIG. 2. Effect of tunicamycin on the expression of CPG2(surface) in NIH3T3 cells FIG. 2A shows (A) Immunoblot analysis. NIH3T3 cells were transfected with either MLVCPG2(surfaceF/S) (lanes 1, 3) or MLVCPG2(surface) (lanes 2, 4). The cells were incubated in the absence of tunicamycin (lanes 1, 2) or in the presence of tunicamycin (lanes 3, 4). Extracts were prepared and analysed as described in FIG. 1. The position of migration of CPG2(surface) is indicated, as is the position of migration of standard molecular weight markers (in kDa), to the left of the figure. FIG. 2B shows (B) CPG2 enzyme activity assay. The enzyme extracts from the cells in part (A) were analysed for CPG2 enzyme activity. Sample numbers correspond to lane numbers for part (A); sample 5 contains extraction buffer instead of cell extract as a negative control. The activity is expressed as arbitrary values relative to buffer controls.
FIG. 3. Effect of glycosylation mutations on expressed CPG2(surface) FIG. 3A shows (A) immunoblot analysis. NIH3T3 cells were tr
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Marais Richard
Springer Caroline Joy
Campell Bruce R.
Cancer Research Campaign - Technology Limited
Nguyen Dave Trong
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