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Pharmaceutical compositions and methods useful for modulating angiogenesis and inhibiting metastasis and tumor fibrosis

a technology of angiogenesis and compositions, applied in the direction of drug compositions, immunoglobulins against animals/humans, cardiovascular disorders, etc., can solve the problems of inability to use unmodified oligonucleotides, interference with correct splicing, and limited solid tumor growth

Inactive Publication Date: 2003-06-19
TECHNION RES & DEV FOUND LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The growth of solid tumors is limited by the availability of nutrients and oxygen.
Alternatively, such hybrid formation may lead to interference with correct splicing.
Unmodified oligonucleotides are typically impractical for use as antisense sequences since they have short in vivo half-lives, during which they are degraded rapidly by nucleases.
Furthermore, they are difficult to prepare in more than milligram quantities.
In addition, such oligonucleotides are poor cell membrane penetrants.
In addition, in order to improve half-life as well as membrane penetration, a large number of variations in polynucleotide backbones have been done, nevertheless with little success.
Such molecules bind to the targeted gene molecules in RNA of tumor cells, thereby inhibiting the translation of the genes and resulting in dysfunctional growth of these cells.
However, due to their low stability RNA oligonucleotides are typically expressed inside the cells using vectors designed for this purpose.
It is possible that some of these diseases result from reduced or abolished lysyl oxidase activity which leads to the synthesis of a fragile extracellular matrix, and consequently, fragile blood vessels.

Method used

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  • Pharmaceutical compositions and methods useful for modulating angiogenesis and inhibiting metastasis and tumor fibrosis
  • Pharmaceutical compositions and methods useful for modulating angiogenesis and inhibiting metastasis and tumor fibrosis
  • Pharmaceutical compositions and methods useful for modulating angiogenesis and inhibiting metastasis and tumor fibrosis

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example 1

The role of LOR1 in Angiogenesis

[0173] A study was conducted in efforts to further substantiate and characterize the role of LOR-1 in angiogenes is.

[0174] Materials and Methods

[0175] Human recombinant platelet factor-4 (PF4, GenBank Accession number M20901) which was produced in bacteria and subsequently refolded was supplied by Dr. Maione of Repligen Corp. (Boston, USA). Estrogen slow release pellets were obtained from Innovative Research of America, Sarasota, Fla., USA.

[0176] Construction of LOR-1 expression vector, transfection into MCF-7 cells, and expression: The LOR-1 cDNA (SEQ ID NO: 1) was cloned into a pCDNA3.1-hygro expression vector (Invitrogen Inc., USA) under the control of a CMV promoter. Clones of cells expressing LOR-1 were selected using hygromycine and assayed for LOR-1 expression using the polyclonal antisera described below.

[0177] Construction of platelet factor-4 affinity columns and purification of LOR-1 on such columns: PF4 was coupled to sepharose using a mod...

example 2

MCF-7 Breast Cancer Cells Expressing Recombinant Lysyl Oxidase Related Protein-1 (LOR-1) form Invasive Tumors Characterized by Extensive Fibrosis

[0200] A study was conducted in efforts to further substantiate and characterize the role of LOR-1 in inhibiting metastasis and tumor fibrosis.

[0201] Materials and Methods

[0202] Estrogen pellets (17.beta.-estradiol, 0.72 mg / pellet, 60-days release) were from Innovative Research of America, FL, USA. Masson Trichrome stain kit was purchased from Bio-Optica (Milano, Italy), Reverse Transcriptase, G418 were from GIBCO BRL (U.K.), Hygromycin B, Tetracycline hydrochloride, Reticulum stain kit fast green and Sirius red (direct red 80) were from Sigma (USA) , Restriction enzymes, T4 ligase were from New England Biolabs (USA). The bacterial expression vector pQE-30 and the nickel affinity column were obtained from Qiagen (Germany) , .sup.32p-dATP was purchased from NEN (USA). Monoclonal anti cytokeratin-7 (CAM 5.2) coupled to FITC was acquired from ...

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Abstract

Methods and compositions suitable for modulating angiogenesis in a mammalian tissue are provided. Further provided are methods suitable for inhibiting metastasis and fibrosis in a mammalian tissue.

Description

[0001] This is a Continuation-In-Part of PCT / IL01 / 00728, filed Aug. 7, 2001, which claims the benefit of priority from U.S. Provisional Patent Application No. 60 / 223,739, filed Aug. 8, 2000.FIELD AND BACKGROUND OF THE INVENTION[0002] The present invention relates to pharmaceutical compositions and methods useful for modulating angiogenesis and for inhibiting metastasis and fibrosis in a mammalian tissue.[0003] Angiogenesis[0004] In an adult, formation of new blood vessels in normal or diseased tissues is regulated by two processes, recapitulated vasculogenesis (the transformation of pre-existing arterioles into small muscular arteries) and angiogenesis, the sprouting of existing blood vessels (which occurs both in the embryo and in the adult).[0005] The process of angiogenesis is regulated by biomechanical and biochemical stimuli. Angiogenic factors such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are released by vascular cells, macrophages...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61KA61K6/00A61K38/44A61K39/395A61K48/00A61P35/00C07K16/30C07K16/40C12N9/06C12N15/53
CPCA61K2039/505C07K16/30C07K16/40C12N9/0022C12Q1/26G01N33/57419G01N33/574G01N2500/00A61K38/00C12Q1/6886C12Y104/03013A61P35/00A61P35/04A61P9/00
Inventor NEUFELD, GERAAKIRI, GALVADASZ, ZAHAVAGENGRINOVITCH, STELA
Owner TECHNION RES & DEV FOUND LTD
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