Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1996-08-13
1999-09-07
Degen, Nancy
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
514 44, C07H 2104, A61K 4800
Patent
active
059489014
DESCRIPTION:
BRIEF SUMMARY
I. FIELD OF THE INVENTION
The invention relates to antisense oligonucleotides which are suitable for inhibiting the expression of aromatase, a process for the production thereof as well as their use.
II. BACKGROUND OF THE INVENTION
Aromatase belongs to the cytochrome p450 enzyme family. Aromatase is the key enzyme in the estrogen biosynthesis. It converts the male sex hormones (androgens) into the female ones (estrogens). The latter are growth factors for a plurality of tumors, particularly those of ovaries, endometrium and breast.
For treating the above tumors, it is tried to inhibit the estrogen biosynthesis. Aromatase inhibitors are often used for this purpose. However, they have not shown satisfactory results by now, particularly they are lacking specificity.
Therefore, it is the object of the present invention to provide a preparation by which aromatase can be inhibited specifically.
According to the invention this is achieved by providing an antisense oligonucleotide which prevents the expression of aromatase by attachment to aromatase DNA and/or mRNA. The expression "antisense" is generally known and refers to a complementarily of the oligonucleotide sequence to the region of aromatase DNA and/or mRNA.
III. SUMMARY OF THE INVENTION
The present invention is directed to an antisense oligonucleotide suitable for inhibiting the expression of aromatase, the antisense oligonucleotide being obtainable by the following steps: coding and regulatory regions of an aromatase DNA and/or transcripts thereof, the antisense oligonucleotides overlapping; antisense oligonucleotides of (a); and well as identification of the antisense oligonucleotide(s) responsible for this.
The present invention also directed to a process for preparing such an antisense oligonucleotides as well to methods for its use.
IV. BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1A depicts a Northern Blot reflecting the effect of an antisense oligonucleotide according to the invention on the expression of aromatase mRNA.
FIG. 1B depicts a densiometric quantification of the Northern Blot shown in FIG. 1A.
FIG. 2A depicts the effect of an antisense oligonucleotide according to the invention on the aromatase activity.
FIG. 2B depicts the effect of an antisense oligonucleotide according to the invention on the aromatase protein.
V. DESCRIPTION OF THE INVENTION
An antisense oligonucleotide according to the invention can be prepared as usual. A process proves to be favorable which comprises the following steps: coding and regulatory regions of aromatase DNA and/or transcripts thereof, the antisense oligonucleotides overlapping; antisense oligonucleotides of (a); and well as identification of the antisense oligonucleotide(s) responsible for this.
An antisense oligonucleotide according to the invention may have differing lengths. Lengths of 20 to 30 nucleotides are preferred. Furthermore, the antisense oligonucleotide may have variations in its sugar and phosphate components each. The sugar component conceivable is deoxyribose, ribose or a chemical variant thereof, for example. The phosphate component may be, e.g., ortho-phosphoric acid diester or a chemical variant thereof. A preferred antisense oligonucleotide contains deoxyribose as sugar component and ortho-phosphoric acid diester as phosphate component.
In the above step (a), antisense oligonucleotides are constructed along the entire length of coding and regulatory regions of an aromatase DNA and/or transcripts thereof. An aromatase DNA and transcripts thereof are known from Mahendroo et al., 1993, J. Biol. Chem..sub.13 :19463-19470, for example. Such a DNA comprises 9 coding exons (exons II-X). Furthermore, several exons I (exons I.1.I.4) exist which are non-coding, transcribed in tissue-specific manner and thus have regulatory functions. The various aromatase transcripts (mRNA) are equal in the translated region (exons II-X) but differ in a tissue-specific manner in the 5' untranslated region (exon I).
Antisense oligonucleotides are constructed as usual. It is favorable to construct them in
REFERENCES:
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Ackermann Karin
Fauss Jurgen
Pyerin Walter
Degen Nancy
Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Recht
Epps Janet
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