Applications of VEGF receptor fusion protein in preparation of medicament for curing diseases about angiogenesis

A fusion protein, angiogenesis technology, applied in sexual diseases, metabolic diseases, skin diseases, etc., can solve the problems of no therapeutic effect, large side effects, and reduced soluble VEGFR2 expression.

Active Publication Date: 2008-10-08
CHENGDU KANGHONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These drugs have no effect on the immunopathological mechanism of RA, have no therapeutic effect on joint destruction, and have no effect on the progress and prognosis of the disease. The side effects are mainly gastrointestinal symptoms
The second class of slow-acting antirheumatic drugs (SAARD) includes reliever drugs (DMARDS) and cytotoxic drugs. This class of drugs has a longer onset time. In addition to improving symptoms, it can also suppress abnormal immune function, but most drugs have major side effects
The third category is glucocorticoids (SAIIDS), which is currently the strongest anti-inflammatory drug, but it cannot block the progression of RA and joint destruction, and has obvious side effects and drug dependence
[0028] The concentration of VEGF and soluble VEGFR1 in the serum of patients with systemic lupus erythematosus is higher than that of normal people, but the expression of soluble VEGFR2 in patients with active systemic lupus erythematosus is reduced (Robak E et al., (2003) MediatorsInflamm, 12(5 ):293-8)

Method used

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  • Applications of VEGF receptor fusion protein in preparation of medicament for curing diseases about angiogenesis
  • Applications of VEGF receptor fusion protein in preparation of medicament for curing diseases about angiogenesis
  • Applications of VEGF receptor fusion protein in preparation of medicament for curing diseases about angiogenesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] The construction of embodiment 1FP7

[0080] The fusion protein FP7 was synthesized by primers Flt-1D2(F) 5'-cctttcgtagagatgtacagtga-3', Flt-1D2(R) 5'-tatgattgtattggtttgtccat-3', KDR D3(F) 5'-gatgtggttctgagtccgtctca-3' and KDR D3 -4(R)5'-cggtgggacatacacaaccaga-3' is recombined from the Flt-1 and KDR gene fragments obtained from the cDNA synthesized from the mRNA extracted from HUVEC cells as a template. The specific conditions are: denaturation at 95°C for 30 minutes, annealing at 56°C for 45 seconds, extension at 72°C for 2 minutes, and PCR amplification for 30 cycles to obtain Flt-1 and KDR IgG-like domains of PCR products. Using TA cloning kit, clone the PCR product into PCR2.1 plasmid, and transfect E.coli, select white colonies, add LB medium, and culture overnight. After the plasmid was extracted with Qiangen plasmid extraction kit, it was digested and sequenced for identification. The splicing PCR (Sewing PCR) method is used to connect the Flt-1 fragment, the ...

Embodiment 2

[0081] The construction of embodiment 2 FP8

[0082] The fusion protein FP8 is directly amplified by PCR using FP7 as a template, and the primer used in PCR is flt-1D 2(F) 5'-cctttcgtagagatgtacagtga-3' and KDR D3-hing (R). The nucleotide sequence of the latter is: 5'-aggtgctgggcacagtgggcatgtgtgagttttgtctttttcatggaccctgacaaatg-3'. It includes a sequence complementary to the third immunoglobulin-like region of KDR and a partial nucleotide sequence of the human IgG Fc hinge region. The method of PCR amplification and gene cloning is identical with example 1. Finally, the PcDNA3.1 plasmid inserted with FP8 was transfected into CHO cells, and stable cell beads were obtained for protein expression.

Embodiment 3

[0083] The effect of embodiment 3 fusion protein FP3, FP7 in mouse rheumatoid arthritis

[0084] The CIA mouse model is the most widely used animal model of rheumatoid arthritis, and the model is established by immunizing mice with bovine collagen (CII). After mixing 2 g / L of CII and complete Freund's adjuvant at a ratio of 1:1, inject intradermally at the root of the tail of DBA / 1J mice (8-10 weeks), 200 μL each. After 21 days, the same concentration of CII and complete Freund's adjuvant were injected at the same site for a second immunization, 200 μL each. The establishment of the CIA model was evaluated according to the degree of erythema, swelling and joint deformation in the ankle and toe joints. The degree of joint swelling was evaluated by measuring the thickness of the foot with a thickness measuring ruler: 0 points: normal joint; 1 point: mild joint swelling with or without erythema, no joint deformation; 2 points: obvious joint swelling, but no joint deformation; 3 ...

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Abstract

The invention relates to an application of the fusion proteins FP1-FP8 of VEGF receptors on the treatment of the diseases relating to angiogenesis. The diseases relating to the angiogenesis includes autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, vasculitis, systemic sclerosis and other diseases caused by the angiogenesis such as bronchial asthma, obesity, nasal polyps, rhinitis, liver cirrhosis, endometriosis, uterine bleeding, warts, scars and ovarian cysts, etc.

Description

technical field [0001] The present invention relates to a series of proteins that can effectively inhibit angiogenesis (angiogenesis), specifically, the application of VEGF receptor fusion protein FP1-FP8 in the preparation and treatment of diseases related to angiogenesis, and the diseases related to angiogenesis include Autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, vasculitis, systemic sclerosis and other diseases caused by angiogenesis such as bronchial asthma caused by angiogenesis, obesity, nasal polyps, rhinitis, liver cirrhosis, uterine Endometriosis, uterine bleeding, warts, scars, ovarian cysts, etc. Background technique [0002] Angiogenesis is the process of growing new blood vessels from pre-existing blood vessels. Most of the blood vessels in the adult body are in a quiescent state, and angiogenesis is only seen in a few pathological or physiological states, such as tumors, diabetic retinal disease, arthritis, anemic organs, and endometr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61K45/06A61P37/02A61P29/00A61P17/00A61P15/00A61P13/12A61P11/06A61P11/02A61P5/18A61P5/14A61P3/10A61P3/04A61P1/16A61P1/04A61P1/00
Inventor 俞德超陈川
Owner CHENGDU KANGHONG BIOTECH
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