VEGI, an inhibitor of angiogenesis and tumor growth

An angiogenesis and preparation technology, applied in the fields shown in , can solve the problems of undisclosed angiogenesis inhibition and the like

Inactive Publication Date: 2001-02-28
HUMAN GENOME SCI INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although the protein described in the present application was first described as TNF-γ in EP95903521

Method used

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  • VEGI, an inhibitor of angiogenesis and tumor growth
  • VEGI, an inhibitor of angiogenesis and tumor growth
  • VEGI, an inhibitor of angiogenesis and tumor growth

Examples

Experimental program
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Embodiment approach

[0230] In another embodiment, the present invention relates to a method for detecting the presence or absence of VEGI polypeptide in a sample, so as to diagnose or prognose diseases related to angiogenesis, by using standard methods known in the art, on the surface (i.e. a solid support ) coated with antibodies specific to VEGI polypeptides, such as microtiter plates or membranes (such as nitrocellulose membranes), and the coated surface with serum, tissue or other biological or other biological or Chemical sample exposure allows for diagnostic analysis. The presence or absence of the resulting complex formed between a VEGI polypeptide and an antibody specific for it in a sample can be detected by any general method known in the art, such as fluorescent antibody spectroscopy or colorimetry. The detection method can be used, for example, in diagnosing or prognosing cancer.

[0231] In another embodiment, the present invention relates to a diagnostic kit comprising an antibody ...

Embodiment 1

[0315] Example 1 Bacterial expression and purification of VEGI polypeptide

[0316]An expression vector was constructed, which contained a polynucleotide encoding 38 amino acids of the N-terminus of the putative full-length VEGI-α polypeptide deleted. The DNA sequence encoding the N-terminally deleted VEGI-alpha polynucleotide was amplified with PCR oligonucleotide primers specific for the VEGI-alpha coding sequence. Additional nucleotides that create restriction sites to facilitate cloning are added to the 3' and 5' sequences, respectively. The 5' oligonucleotide primer has the sequence 5'-GCGCCA TGG CTC TGCACT GGG AAC AT-3' (SEQ ID NO: 3) containing an Nco I restriction site followed by 18 nucleotides of the coding sequence. The 3' primer has the sequence 5'-CGC AAG CTTCTA TAG TAA GAA GGC TCC-3' (SEQ ID NO: 4) containing a HindIII restriction site followed by 18 complementary to the last 15 nucleotides of the coding sequence and a stop codon sub-nucleotides. After digesti...

Embodiment 2

[0319] Embodiment 2: clone and express VEGI polypeptide with baculovirus expression system

[0320] The DNA sequence encoding the full-length VEGI-α polypeptide, ATCC NO75927, was amplified using PCR oligonucleotide primers corresponding to the 5' and 3' sequences of the gene. The 5' primer sequence is 5'-GCG CGG ATC CAC CAT GAG ACG CTT TTT AAGCAA AGT C-3' (SEQ ID NO: 15) and contains a Barn HI restriction enzyme site (in bold) followed by VEGI-α 24 nucleotides of the coding sequence (the translation initiation codon "ATG is underlined). The 3' primer sequence is 5'-CGC GTC TAG ACT ATA GTAAGA AGG CTC CAA AGA AGG-3' (SEQ ID NO: 16), and Contain the cleavage site of restriction endonuclease XbaI and be complementary to 22 nucleotides of the 3' untranslated sequence of VEGI-α coding sequence. Use commercially available kit ("Geneclean," BIO 101 Inc., La Jolla, Ca.) The amplified sequence was separated from a 1% agarose gel. This fragment was then digested with endonucleases BamH...

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Abstract

The present invention relates to a vascular endothelial cell inhibitor, VEGI, capable of inhibiting angiogenesis in cellular models and tumourigenesis in animal models, and methods of use.

Description

Background of the invention: [0001] In very limited circumstances, a human or animal under normal physiological conditions undergoes angiogenesis, the creation of new blood vessels in a tissue or organ. For example, angiogenesis is commonly seen in wound healing, embryonic development and formation of the corpus luteum, endometrium and placenta. The term "endothelium" refers to a thin layer of squamous epithelial cells lining the serous cavities, lymphatics, and blood vessels. The term "anti-angiogenic" or "angiogenesis inhibitory activity" refers to the ability of a molecule to inhibit angiogenesis in general. [0002] Controlled and uncontrolled angiogenesis are thought to proceed in a similar manner. Endothelial and mesenchymal cells surrounded by a basement membrane form capillaries. Angiogenesis begins with the erosion of the basement membrane by enzymes released by endothelial cells and leukocytes. Endothelial cells located within the lumen of the vessel then protrud...

Claims

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Application Information

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IPC IPC(8): C12N15/09A61K31/711A61K38/00A61K48/00A61P9/10A61P17/02A61P17/06A61P27/02A61P27/06A61P35/00A61P37/06A61P43/00C07K14/47C07K14/705C12Q1/68
CPCC12N2799/026A61K38/00C07K14/70575A61P1/00A61P17/00A61P17/02A61P17/06A61P27/02A61P27/06A61P35/00A61P37/00A61P37/06A61P43/00A61P9/00A61P9/10A61P9/14Y02A50/30C12N15/62C07K14/515A61K31/7088
Inventor 李鲁远翟一帆余国良倪健马克·E·李普曼克雷格·A·罗森
Owner HUMAN GENOME SCI INC
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