Dual Targeting Drug Carriers
A target and drug technology, applied in the field of novel targeted drug carriers, can solve the problems of tumor cell instability, poor effect, slow survival of cancer patients, etc.
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[0031] The present invention provides a dual target drug carrier, comprising a first target molecule and a second target molecule.
[0032] The target molecule of the present invention is a peptide, an antibody or a protein. The target molecules of the present invention can be chemically modified to increase stability. Chemically modified peptides or peptide analogs include any chemical functional equivalent that increases stability and / or efficacy in vivo (in vivo) or in vitro (in vitro). A peptide analog refers to an amino acid derivative of any peptide. Peptide analogs include, but are not limited to, modification of branched chains at the time of synthesis, incorporation of unsaturated amino acids and / or derivatives thereof, cross-linking, and other methods that increase peptide image constraints. For example, branched chain modifications include modifications of amine groups.
[0033] The target molecule can be bound in vivo and in vitro to lesion tissues, especially t...
Embodiment 1
[0043] 1. Preparation of fusion protein bodies
[0044] The EGF, RGD-EGF, RGD4C-EGF, VEGF, RGD-VEGF and RGD4C-VEGF genes were constructed in the expression vector pET28a(+) with Nco I and Xho I restriction enzymes. The C-terminus (ie, carbon-terminus) of RGD and RGD4C is connected to the N-terminus (ie, nitrogen-terminus) of EGF or VEGF, and the C-terminus of EGF or VEGF has His 6 logo. His 6 The markers are helpful for subsequent purification and analysis, wherein the gene sequences of RGD-EGF, RGD4C-EGF, RGD-VEGF and RGD4C-VEGF contain the linker GG. The constructed pET28a(+)EGF, RGD-EGF, RGD4C-EGF, VEGF, RGD-VEGF and RGD4C-VEGF expression vectors were transfected into E.coli.DH5α to produce and preserve vector DNA. The sequence was confirmed to be correct by DNA sequencing.
[0045] 2. Expression and purification of target fusion protein
[0046] The vector of Example 1 was transferred into E. coli BL21 (DE3) by heat shock, and was screened and confirmed with kanamycin...
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