Peptide with permeable cell membrane or body tissue barrier functions and application of peptide
A cell membrane and barrier technology, applied in the field of biopharmaceuticals, can solve the problems of high cytotoxicity and low transfer efficiency
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[0078] As an embodiment of the present invention, the functional molecule is a preparation with the function of molecular packaging and loading, including but not limited to liposomes, polymers, dendritic molecules, nano-packaging preparations and the like.
[0079] As an embodiment of the present invention, the functional molecule is a viral vector capable of carrying genetic material, including but not limited to a retroviral vector, a lentiviral vector, or an adenoviral vector.
[0080] The connection method between the membrane-penetrating peptide and the functional molecule of the present invention can be covalent connection or non-covalent connection. It should be understood that as long as the functions of membrane-penetrating peptides and functional molecules can be retained, and the effect of penetrating cell membranes and body tissue barriers can be preserved, any connection method can be included in the present invention. Covalent linking usually links two molecules...
Embodiment 1
[0110] Example 1. In vivo directed evolution screening of eye guide peptides
[0111] The present inventors used phage display technology to enrich specific recombinant phages from various variants (peptide library, peptide library) of recombinant phages purposefully by using phage display technology in vivo directed evolution method, and through sequencing analysis, combining the enrichment effect and the Based on the affinity of retinal target tissue, the dominant recombinant phage peptides with the potential to penetrate the eye barrier were screened out.
[0112] In this embodiment, Ph.D. TM The system (M13KE), which is based on the M13 single-chain phage vector (M13mp19), introduces a recombination site at the 5' end of its gene gIII, thereby fusing and recombining the N-terminal of a polypeptide consisting of 12 amino acid residues into the phage minor capsid protein pIII. Each recombinant phage displayed 5 single-cloned polypeptides, and recombinant phages displaying ...
Embodiment 2
[0143] Example 2, Determination of eye barrier permeability of eye guide peptide CC12 and its connecting peptide
[0144] The absorption of foreign substances by the eye has strict selective permeability, and this selection process is mainly realized through the biological barrier of the eye. The blood-retinal barrier is a key biological barrier in the eye, which can selectively regulate the entry of foreign substances into the intraocular tissue, thereby limiting the process of drug delivery from the systemic blood circulation to the posterior segment of the eye. The blood-retinal barrier is composed of an inner barrier and an outer barrier, and its characteristics are similar to those of the blood-brain barrier. Retinal capillary endothelial cells and their connections constitute the inner blood-retinal barrier (iBRB), and retinal pigment epithelium (RPE) and their connections constitute the outer BRB (oBRB).
[0145] Transscleral diffusion delivers drugs to the outer retin...
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