Antisense oligonucleotides for inhibiting helicobacter pylori ac

Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives

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536 231, 536 241, 536 243, 536 2431, C07H 2102, C07H 2104

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059773405

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BRIEF SUMMARY
The present invention concerns antisense oligonucleotides which selectively hybridize with one or more genes necessary for the action of Helicobacter pylori (H. pylori), pharmaceutical compounds comprising them and their use as Helicobacter pylori inhibitors.
Helicobacter pylori (H. pylori) is a microaerophilic bacterium, gram negative, colonizing the intercellular interstices and junctions of the human gastric mucous membrane and establishing a chronic infection with numerous different clinical manifestations such as atrophic gastritis, peptic and duodenal ulcer, gastric atrophy, and gastric carcinoma. The numerous clinical isolated substances have permitted classification of H. pylori into two groups based on the presence or the absence of vacuolizing cytotoxin. In vitro experiments have shown that the virulent nature of the bacterium may be connected to its mobility and to the presence of vacuolizing cytotoxin. Likewise, there is a direct relationship between the expression of cytotoxin and the presence of an immuno-dominant CagA antigen exposed on the surface. Thus, inhibition of the mobility of the bacterium and/or expression of the cytotoxic factor may prevent the manifestation of clinical symptoms. One of the ways to inhibit these factors comprises using antisense oligonucleotides to block the expression of the coding of the genes for the immuno-dominant antigen associated with the CagA cytotoxicity and/or flagellin (flaA and flaB) and/or the vacuolizing cytotoxin (vacA).
The antisense strategy is a therapeutic approach whose purpose is the selective modulation of the expression of the genes by a highly selective association of a chain of nucleotides (oligonucleotides) with its supplementary sequence on RNA or DNA and consequently the inhibition of the synthesis of the corresponding protein.
The oligonucleotides complementary to the products of transcription are called "antisense" oligonucleotides. Nucleotides having the same sequence as the products of transcription are called "sense" oligonucleotides. Initially, these compounds were logically intended to inhibit the formation of a product of the gene by suppression of the corresponding messenger RNA via the hydrolysis mechanism catalyzed by RNAse H. It soon turned out that the mechanism of action of these antisense oligonucleotides was not so simple. These oligonucleotides may interact with a certain number of cell targets not containing nucleic acid. These oligonucleotides may interact with the gene to form triple helix structures and inhibit the formation of products of transcription. Oligonucleotides may interact with the intron-exon junctions of pre-messenger RNA, thus interfering with the correct splicing of the product of transcription. Oligonucleotides may hybridize with messenger RNA in the cytoplasm forming an RNA-DNA complex which is quickly degraded by the RNAas H enzyme or by impeding the ribosome complex from splicing onto the messenger RNA and thus blocking the translation. Oligonucleotides, and more especially modified oligonucleotides, may interact with a number of cellular products such as proteins. These interactions may be sequence specific (for example: transcription factors) or non-sequence specific (for example: growth factors).
Oligonucleotides are very often used as a probe, for example for the detection or isolation of a polynucleotide and/or as a primer for the transcription and/or replication of target sequences (N. A. Innis et al., PCR Protocols: A Guide to Methods and Applications, 1990, Academic Press, Inc., San Diego, Calif.). Thus it is that oligonucleotides have been used during studies on Helicobacter pylori (WO-A-93/18150) and more especially on vacuolizing cytotoxin vacA (S. Phadnis et al, Infection and Immunity, vol. 62, No. 5, 1994, pp. 1557-1565) or the CagA antigen (M. Tummuru et al., Infection and Immunity, vol. 61, Nos. 5, 3, Washington, DC, pp. 1799-1809, and A. Covacci et al., Proceedings of the National Academy of Sciences of the USA, vol. 90, No. 12, 1993, Washington, DC, pp. 5791-5795) via the PCR (po

REFERENCES:
patent: 5527678 (1996-06-01), Blaser et al.
Uhlmann et al. "Antisense Oligonucleotides: A New Therapeutic Principle" Chemical Reviews. vol. 90(4): 543-584, Jun. 1990.
Agrawal, S. "Antisense Oligonucleotides: Towards Clinical Trials" TIBTECH vol. 14:376-387, Oct. 1996.
Branch, D. "A Good Antisense is Hard to Find" TIBS vol. 23:45-50, Feb. 1998.
Telford et al, "Unravelling . . and Vaccines", Trends in Biotechnology vol. 12, No. 10, Oct. 1, 1994, pp. 420-456.
G. Zon, "Oligonucleotide . . . Agents", Pharmaceutical Reseach, vol. 5 (1988) , pp. 539-549.
Phadnis et al, "pathological . . . Helicobacter pylori", Infection and Immunity, vol. 62, No. 5, 1994 pp. 1557-1565.
Tummuru et al, "Infection and Immunity", vol. 61, No. 5, (1993), pp. 1799-1809.

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