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Polypeptide, polypeptide nano drug-loading carrier and preparation method of polypeptide and polypeptide nano drug-loading carrier

A nano-drug loading and carrier technology, applied in the field of bioengineering, can solve problems such as unsatisfactory splitting conditions, and achieve the effects of avoiding potential biological toxicity and immunogenicity, low synthesis cost and good stability.

Inactive Publication Date: 2019-11-08
WEIFANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The drug-loaded carriers in the prior art have a certain degree of potential harm after decomposition, while the polypeptide does not have any potential reaction after decomposition, and the splitting conditions of the drug-loaded carriers in the prior art cannot be satisfied in the tumor area. Therefore, it is possible to combine the characteristics of amphiphilic peptides to prepare a drug-loaded carrier targeting the tumor area to meet people's needs.

Method used

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  • Polypeptide, polypeptide nano drug-loading carrier and preparation method of polypeptide and polypeptide nano drug-loading carrier
  • Polypeptide, polypeptide nano drug-loading carrier and preparation method of polypeptide and polypeptide nano drug-loading carrier
  • Polypeptide, polypeptide nano drug-loading carrier and preparation method of polypeptide and polypeptide nano drug-loading carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1: Preparation of Ac-LLLLLLKKKK-NH2 polypeptide

[0060] Step 1. Distill N,N-dimethylformamide (DMF) and piperidine (Piperidine) solvents

[0061] Distill the purchased DMF solution under reduced pressure at 60°C to obtain pure DMF solvent; add a small amount of CaH2 to the purchased piperidine and heat to reflux for 1-2 hours, and receive the fraction with boiling point temperature (106°C) to obtain pure piperidine pyridine solvent.

[0062] Step 2, preparation of amino acid, resin, activator, capping agent, deprotecting agent

[0063] The amount of amino acids and other reagents required to prepare 0.25mM NH2-AAAAAAKK-NH2 is calculated on the polypeptide solid-phase synthesizer:

[0064] Leu (leucine): 1.97g dissolved in 21mL DMF;

[0065] Lys (lysine): 2.27g dissolved in 32mL DMF;

[0066] Resin (loaded at 0.6mmol / g): 0.417g;

[0067] Washing: TFA: 142mL;

[0068] Deprotection solution: piperidine: 48mL; DMF: 192mL (20% piperidine / 80% DMF);

[0069]...

Embodiment 2

[0075] Example 2: Preparation of Ac-LLLLLLKKKK-NH2 drug-loaded carrier

[0076] Weigh 1.25mg (1mmol / L) polypeptide Ac-LLLLLLKKKK-NH2, add 1mL of Hepes buffer solution, sonicate for 5min, and let it stand at room temperature for 1h or 6 months. SEM and TEM observation results show that stable nanoparticles are formed at this time. Assembly ( figure 1 , Figure 4 ).

Embodiment 3

[0077] Example 3: Detection of self-assembled morphology of Ac-LLLLLLKKKK-NH2 drug-loaded carrier in Hepes buffer

[0078] The specific detection method is as follows:

[0079] Examination of self-assembled morphologies (TEM, SEM) in Hepes buffer.

[0080] SEM: The prepared peptide solution was lyophilized into powder using a vacuum freeze dryer (EYELAFDU-1100, Japan) to obtain sample powder. On a scanning electron microscope (SEM, HITACHI, SU8010, Japan), the sample was observed at a voltage of 3 kV after spraying gold. The results showed that the peptide sample Ac-LLLLLLKKKK-NH2 observed by scanning transmission electron microscopy self-assembled into a uniform nanosphere structure in Hepes buffer, as Figure 5 shown.

[0081] TEM: Transmission electron microscopy (TEM) measurements were performed on a HITACHI HT7700 instrument (Japan) at an accelerating voltage of 80 kV. Take the peptide solution obtained in Example 2 and let it stand for a specific time, or adjust a sp...

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Abstract

The invention discloses a polypeptide, a polypeptide nano drug-loading carrier and a preparation method of the polypeptide and the polypeptide nano drug-loading carrier. The polypeptide has a structural formula of Ac-LLLLLLKKKK-NH2. The preparation method of the polypeptide comprises the steps that various raw materials are added to a polypeptide solid phase synthesizer, and Ac-IIIIKK-NH2 is synthesized from a C-terminal to an N-terminal; and Ac-IIIIKK-NH2 is further prepared to obtain the requried polypeptide. The polypeptide nano drug-loading carrier is of a nanosphere structure capable of being ruptured in an acidic environment, and the nanosphere structure is used for embedding a fat-soluble drug. The preparation method of the polypeptide nano drug-loading carrier comprises the steps that the polypeptide is added to a Hepes solution, and the polypeptide is self-assembled to form the polypeptide nano drug-loading carrier embedding the fat-soluble drug.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a polypeptide, a polypeptide nanometer drug-carrying carrier and a preparation method for the two. Background technique [0002] Peptide self-assembly is a spontaneous thermodynamic process that requires the cooperative participation of multiple intermolecular non-covalent interactions, ultimately determining the self-assembly morphology in a thermodynamically stable minimum energy state. These non-covalent interactions include hydrogen bonding interactions that form stable structures between peptides and proteins, π-π interactions that induce directional growth of molecules, electrostatic interactions after ion pair formation, and play an important role in interfacial systems. Hydrophobic interactions and van der Waals forces. Meanwhile, the self-assembly process can be precisely and rationally regulated by various kinetic parameters, such as pH, temperature, ions, concentrations,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/04C07K1/14A61K9/51A61K47/42
CPCA61K9/5169C07K7/06
Inventor 白靖琨龚中英
Owner WEIFANG MEDICAL UNIV
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