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Novel therapeutic protein and target e2-epf5

A technology of E2-EPF5 and ubiquitin, which is applied in the field of new therapeutic proteins and targets E2-EPF5, can solve the problems that the understanding of ubiquitin system relationship is in its infancy

Inactive Publication Date: 2007-09-19
NOVARTIS AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the availability of human genome sequences has made it possible to complete the identification of members of the ubiquitin family, including E2, E3, and deubiquitinating proteases, only a few ubiquitinating substrates have been identified, and the ubiquitin system and its association with The understanding of human disease relationships is still in its infancy

Method used

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Embodiment

[0058] 1. Antisense (ASO) and siRNA oligonucleotides

[0059] siRNA is available from a number of commercial suppliers, eg Qiagen, Dharmacon, Proligo.

[0060] Oligoribonucleotides were synthesized using TOM-phosphoramidite chemistry and purified by RP-HPLC as described by the manufacturer (Xeragon). Purity was assessed by capillary gel electrophoresis. Quantification was performed by UV based on an extinction coefficient of 260 nM. Annealing of double-stranded RNA (dsRNA) was performed as described elsewhere (Elbashir et al., 2001).

[0061] Modified antisense oligodeoxynucleotides were synthesized using phosphoramidite chemistry and purified by HPLC, analyzed and characterized by electrospray mass spectrometry and capillary gel electrophoresis. Quantification was performed by UV based on an extinction coefficient of 260 nM.

[0062] All oligonucleotides were designed against the human E2-EPF5 sequence (GenBank accession number M91670, GI number: GI: 181915, SwissProt ent...

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Abstract

The present invention relates to novel uses for ubiquitin conjugating enzyme E2-EPF5. In particular, inhibition of E2-EPF5 activity is shown to reduce the production VEGF, as well as other proteins regulated the transcription factor HIF-1, in response to hypoxia. Based on these findings the present invention provides therapeutic methods and pharmaceutical compositions useful for the treatment of diseases related VEGF induced angiogenesis. In addition, E2-EPF5 is provided as target for the development of therapeutics. Accordingly, the invention provides screening methods for identifying candidate compounds that inhibit E2-EPF5 activity and may therefore be used to treat VEGF induced angiogenesis such as tumor related angiogenesis. Finally, the invention also provides methods for the inhibition of VEGF and other HIF-1 regulated proteins.

Description

[0001] screening method technical field [0002] The present invention relates to therapeutic methods and pharmaceutical compositions for inhibiting VEGF-dependent angiogenesis, particularly tumor-associated angiogenesis. Background technique [0003] Vascular endothelial growth factor (VEGF) encodes a protein that responds to a variety of vascular responses observed in human tumors, including stimulation of neovascular protrusion and its permeabilization. VEGF is essential for the generation of angiogenesis in most solid tumors, and constitutive upregulation of VEGF expression is considered a major contributor to tumor angiogenesis. [0004] The expression of VEGF and several other angiogenic growth factors is tightly regulated by hypoxia, involving the transcriptional induction of the VEGF gene by the transcription factor HIF-1 (hypoxia-inducible factor-1). Hypoxia is a reduction in the normal level of tissue oxygen tension homeostasis and occurs in acute and vascular dis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395G01N33/50
CPCC12N2310/346C12Y603/02019C12N2310/321C12N2310/14C12N2310/341C07K16/40C12N2310/315C12N2310/11C12N2310/3341C12N15/1137A61P1/16A61P17/06A61P19/02A61P27/02A61P35/00A61P43/00C12N2310/3525A61K31/7105A61K39/395G01N33/50
Inventor F·A·阿塞尔贝格J·哈尔D·许斯肯M·A·拉博C·S·米克宁P·施密特J·魏勒L·怀德尔
Owner NOVARTIS AG
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