Conjugate of VEGF-GRAB protein and drug, and use thereof

A technology of conjugates and drugs, applied in 3 fields, can solve the problems of not having the function of targeting tumor cells, not showing the anti-cancer effect of cancer cells, harmful to normal blood vessels, etc.

Pending Publication Date: 2020-04-17
KOREA ADVANCED INST OF SCI & TECH
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, conventional angiogenesis inhibitors are useful in the treatment of cancer because they inhibit angiogenesis required for the proliferation of cancer cells, but such inhibitors do not have the function of targeting tumor cells, and thus cannot exhibit cancer cell-specific anti-tumor activity. Carcinogenic and deleterious effects on normal blood vessels

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Conjugate of VEGF-GRAB protein and drug, and use thereof
  • Conjugate of VEGF-GRAB protein and drug, and use thereof
  • Conjugate of VEGF-GRAB protein and drug, and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] Example 1: Cell lines and cell culture

[0110] Freestyle 293F cells (R790-07, ), A431 cells (human cervical epidermoid carcinoma, #21555, Korean Cell Line Bank), SKBR3 cells (human breast cancer, #30030, Korean Cell Line Bank), SKOV3 cells (human ovarian adenocarcinoma, #30077, ATCC) and Human umbilical vein endothelial cells (HUVEC, CC-2519, Lonza) were certified according to ATCC guidelines and used within 6 months of receipt. at 37°C and 8% CO 2 Next, Freestyle 293F cells (R790-07, ) maintained in Freestyle293F medium (12338018, ) in suspension culture with 125rpm stirring. A431 cells were cultured in DMEM (LM001-05, Welgene) supplemented with 10% heat-inactivated FBS (S001-01, Welgene) and 100 μg / ml penicillin / streptomycin, and SKBR3 cells and SKOV3 cells were supplemented with 10% heat-inactivated FBS (S001-01, Welgene) and 100 μg / ml of penicillin / streptomycin RPMI1640 (LM011-05, Welgene) were cultured, and HUVECs were incubated in gelatin (G9391, Sigma-...

Embodiment 2

[0111] Example 2: Antibodies

[0112] Antibodies useful in the present invention are shown in Table 1 below.

[0113] [Table 1]

[0114]

Embodiment 3

[0115] Embodiment 3: Expression and purification of recombinant protein

[0116] The gene encoding the cetuximab or trastuzumab single-chain variable fragment (scFv) (where the variable regions of the cetuximab or trastuzumab heavy and light chains pass through the (G4S)3 linker Connection (Ahmad ZA, Clin Dev Immunol.2012, 2012:980250.)) and VEGF-Grab's N-terminal connection (Lee JE, Mol Cancer Ther., 2015, 14:470-9) (attended figure 1 A). Polyvinylamine (765090, Sigma-Aldrich) containing VEGF-Grab, scFv-cetuximab-VEGF-Grab (Cet-Grab) and scFv-trastuzumab-VEGF-Grab (Tras-Grab) The vector was transfected into Freestyle293F cells. Transfected cells were cultured with 5 mM sodium butyrate (303410, Sigma-Aldrich) for 3 days, and then centrifuged using a centrifuge to separate only the supernatant. Supernatants containing VEGF-Grab, Cet-Grab, or Tras-Grab were purified using Protein A Sepharose (GE Healthcare LifeSciences). VEGF-Grab, Cet-Grab, or Tras-Grab were eluted with 2...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention relates to a VEGF-Grab protein-drug conjugate and a use thereof and, more particularly, to a conjugate of a fusion protein in which a VEGFR1 domain 2, a VEGFR1 domain 3, and an antibody fragment are connected and a drug, a pharmaceutical composition for prevention or treatment of cancer or angiogenesis-related disease, comprising the conjugate, and a method for prevention ortreatment of cancer or angiogenesis-related disease. Serving as a multi-paratopic VEGF decoy receptor, the conjugate including a VEGF-Grab protein and a drug of the present invention can be used as amulti-purpose platform for treatment of cancer or angiogenesis-related disease.

Description

technical field [0001] The present invention relates to a conjugate of a VEGF-Grab protein and a drug and its use, more particularly to a conjugate of a fusion protein and a drug in which VEGFR1 domain 2, VEGFR1 domain 3, and antibody fragments are connected to each other. A pharmaceutical composition for preventing or treating cancer or angiogenesis-related diseases comprising the conjugate, and a method for preventing or treating cancer or angiogenesis-related diseases. Background technique [0002] For the proliferation and growth of cancer cells, new blood vessels that supply oxygen and nutrients are required, and it is known that vascular endothelial growth factor (VEGF) plays a key role in the formation of new blood vessels. VEGF is an approximately 46 kDa dimer composed of two subunits, and five types of VEGF (VEGF-A, VEGF-B, VEGF-C, VEGF-D, and PlGF) are currently known in mammals. VEGF binds to three receptor tyrosine kinases (RTKs) called VEGF receptors (VEGFR)-1,...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/64C07K14/71C07K16/00A61K39/395A23L33/10A23K20/10
CPCA23K20/10A23L33/10A61K39/395C07K14/71C07K16/00A61K47/6425A61K47/68A61K47/64A61K39/3955C07K2319/00C07K2317/52A23V2002/00A23V2200/308A61K45/06C07K14/475C07K16/22C07K16/28
Inventor 金昊民李大熙金德起
Owner KOREA ADVANCED INST OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap