Ligand polypeptide specifically combined with MDSCs (Myeloid-Derived Suppressor Cells) and drug delivery system

A delivery system and specific technology, applied in the field of protein and polypeptide, can solve problems such as unsatisfactory therapeutic effect and complexity of immune regulation network, and achieve good gene silencing effect, good binding effect, and good binding effect

Active Publication Date: 2014-03-12
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, due to the heterogeneity of MDSCs itself and the complexity of the immune regulatory network, the above methods have not achieved satisfactory therapeutic effects, so an in-depth understanding of the occurrence and development of MDSCs is very necessary to completely reverse the immunosuppressive activity of MDSCs

Method used

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  • Ligand polypeptide specifically combined with MDSCs (Myeloid-Derived Suppressor Cells) and drug delivery system
  • Ligand polypeptide specifically combined with MDSCs (Myeloid-Derived Suppressor Cells) and drug delivery system
  • Ligand polypeptide specifically combined with MDSCs (Myeloid-Derived Suppressor Cells) and drug delivery system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. In vivo phage screening technology to screen candidate polypeptides for targeting ligands of MDSCs

[0047] The MDSCs-targeting polypeptide DPPWLSW (F7 for short, as shown in SEQ ID NO. 1) was obtained by three rounds of screening using in vivo phage screening technology. The specific method is as follows:

[0048] (1) In vivo phage technology screening

[0049] First round of screening:

[0050] 1. Take 1 tumor-bearing mouse and inject 100 μl Ph.D.-7 into the tail vein TM Phage 7 peptide library (4*10 11 pfu). After 1 h, 4% μl of chloral hydrate and 200 μl of chloral hydrate were anesthetized, and 40 ml of PBS was perfused in the myocardium, and then the tumor tissue was collected.

[0051] 2. Lyse MDSCs cells: add the sorted MDSCs cells to 100 μl of 1% NP-40 cell lysis solution, mix well, and lyse on ice for 5 min.

[0052] 3. A single colony of ER2738 was inoculated into LB-Tet (LB-tetracycline) medium for overnight culture, and the culture was dilu...

Embodiment 2

[0083] Example 2. Synthetic materials with MDSCs targeting

[0084] This embodiment relates to a synthetic material with MDSCs targeting properties, and the synthetic material is obtained by linking a derivative polypeptide of the MDSC molecule-specific targeting polypeptide with Mal-PEG2000-DSPE. When the sequence of the specific targeting polypeptide of the MDSCs molecule is synthesized, GGGC is added to the C-terminus to obtain a derivative polypeptide whose sequence is DPPWLSWGGGGC (as shown in SEQ ID NO. 3); the cysteine ​​of the derivative polypeptide is free The sulfhydryl group reacts with the maleimide group and is connected to DSPE-2000-Maleimide (distearoyl glycerol phospholipid ethanolamine polyethylene glycol 2000 maleimide, and polyethylene glycol can also choose PEG5000) Above, a synthetic material with MDSCs targeting is obtained; further, the MDSCs-specific targeting polypeptide can also be directly connected with DSPE-2000-Maleimide; another synthetic mater...

Embodiment 3

[0090] Example 3. Derivative polypeptides of MDSCs targeting polypeptides

[0091] 1. Binding experiment of MDSCs, Jurkat and polypeptide DPPWLSW-FITC (suspended cells)

[0092] 1) MDSCs, Jurkat cell count, 1640 medium (10% fetal bovine serum) to adjust the cell concentration to 2 × 10 6 pcs / ml. 24-well plate, 200 μl of cell suspension per well.

[0093] 2) Under the dark condition, 20 μl of polypeptide DPPWLSW-FITC solution with different concentrations was added to each well, and the concentrations were 1 mM, 500 μM, 50 μM, 5 μM and 0.5 μM respectively. After mixing, 37℃, 5%CO 2 Incubate in the dark for 1 h.

[0094] 3) After the incubation, discard the cell culture medium containing fluorescent peptides, add 400 μl / well of PBS+2% FBS buffer, 300 g, 10 min, centrifuge and wash 3 times, and thoroughly wash to remove free or non-specifically bound fluorescent peptides.

[0095] 4) Using a flow cytometer, count 10,000 cells under the excitation light channel of 488 nm, a...

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Abstract

The invention discloses a ligand polypeptide specifically combined with MDSCs (Myeloid-Derived Suppressor Cells) and a drug delivery system. The ligand polypeptide specifically combined with MDSCs comprises an amino acid sequence as shown in SEQ ID NO.1. The drug delivery system disclosed by the invention comprises the ligand polypeptide, a drug carrying system and at least one active or developing substance, wherein the drug carrying system is specifically lipidosome; the at least one active substance is any substance which is needed to be delivered to a special in-vivo position. The ligand polypeptide disclosed by the invention has good MDSCs-targeting effect, and can be combined with lipidosome to be used as a targeting drug carrier, so that a targeting drug delivery system for tumors is developed.

Description

technical field [0001] The present invention relates to the technical field of protein polypeptides, in particular to a ligand polypeptide that specifically binds to MDSCs and a drug delivery system. Background technique [0002] Tumor is one of the main reasons that affect human health. Theoretically, as the non-self of the body, the immune system can recognize and inhibit the occurrence and development of tumors, but in fact, tumor hosts usually have low immunity and cannot produce effective anti-tumor immunity. In recent years, the tumor immunoediting hypothesis about the interaction between tumor development and the immune system has attracted people's attention. This hypothesis believes that tumor cells can form a stable tumor immune tolerance pattern by reducing their own immunogenicity, down-regulating the expression of co-stimulatory molecules on the cell surface, releasing a large number of immunosuppressive factors, and recruiting a variety of immunosuppressive ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K47/42A61K48/00A61K41/00A61K49/22A61K51/08A61K9/127A61P35/00
Inventor 徐宇虹司晓菲吴烈宜张金平陈晓龙
Owner SHANGHAI JIAO TONG UNIV
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