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Tumor microenvironment responsive surface charge reversible nano-drug delivery carrier

A tumor microenvironment and nanocarrier technology, which is applied in the field of surface charge reversible nano-drug delivery carriers, can solve the problems of non-invasive, reproducible, and high-accuracy detection of tumors, and achieve efficient tumor diagnosis and treatment. Effect, wide application prospect, low economic cost effect

Active Publication Date: 2020-08-07
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, it is still difficult to detect tumors non-invasively, reproducibly, and with high accuracy during or after clinical treatment of tumors, so specific imaging detection technologies are urgently needed to guide tumor treatment

Method used

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  • Tumor microenvironment responsive surface charge reversible nano-drug delivery carrier
  • Tumor microenvironment responsive surface charge reversible nano-drug delivery carrier
  • Tumor microenvironment responsive surface charge reversible nano-drug delivery carrier

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0031] Experimental Example 1 Synthesis of the amphiphilic molecule OMPE involved in the present invention

[0032] (1) Experimental materials

[0033] N-methylmorpholine (NMM), piperidine, trifluoroacetic acid (TFA), dichloromethane (DCM), ninhydrin, vitamin C, phenol, benzotriazole-N,N,N',N '-tetramethyluronium hexafluorophosphate (HBTU), diisopropylethylamine (DIPEA), triisopropylsilane (TIS), N,N-dimethylformamide (DMF), anhydrous ether, Wang-resin, methanol, various Fmoc-protected amino acids, oleic acid, peptide synthesis tubes, shaker, vacuum water pump, rotary evaporator, the above reagents and materials were obtained from commercial sources.

[0034] (2) Solvent preparation

[0035] Reaction solution (ACT) - N-methylmorpholine: N, N dimethylformamide (1: 24)

[0036] Deprotection solvent - piperidine: N, N dimethylformamide (1:4)

[0037] Lysis solution—trifluoroacetic acid (95%), triisopropylsilane (2.5%), ultrapure water (2.5%).

[0038] Ninhydrin detection sol...

experiment example 2

[0050] Experimental example 2 Construction of functional nanocarrier O-NP

[0051] (1) Experimental materials

[0052] (2,3-Dioleoyl-Propyl)-Trimethylamine (DOTAP), Dioleoyl Lecithin (DOPC), Soybean Lecithin (SPC), Cholesterol (CHO), Distearoyl Phosphatidylethanolamine-Polyethylene Diol 2000 (DSPE-PEG 2000 ), methanol, chloroform, phosphate buffered saline (PBS).

[0053] (2) Preparation of O-NPs

[0054] Nano-liposome O-NP was prepared by film dispersion method, and the specific steps were as follows:

[0055] Accurately weigh 1mg DOTAP, 2mg DOPC, 1mg SPC, 0.5mg DSPE-PEG 2000 , 0.5mg CHO and 0.5mgOMPE were dissolved in 5mL of solvent (chloroform / methanol, volume ratio 2:1), and after they were completely dissolved, they were placed in a round-bottomed flask. The temperature is 39°C, and the rotary evaporator rotates slowly and evenly. After the organic solvent volatilizes, a uniform film is formed and attached to the wall of the flask. Inert gas removed residual organic...

experiment example 3

[0058] Experimental Example 3 The uptake of O-NP by cells

[0059] (1) Preparation of imaging preparation (O-NP-Cy2): According to the preparation method of O-NP, accurately weigh 1mg DOTAP, 2mgDOPC, 1mg SPC, 0.5mg DSPE-PEG 2000 , 0.5mg CHO and 0.5mg OMPE, 0.5mg Cy2, dissolved in 5mL of solvent (chloroform / methanol, volume ratio 2:1), after completely dissolved, placed in a round bottom flask. The temperature is 39°C, and the rotary evaporator rotates slowly and evenly. After the organic solvent volatilizes, a uniform film is formed and attached to the wall of the flask. Inert gas removed residual organic solvent in the flask. Add 5 mL of PBS, heat and hydrate in a water bath at 60°C for 15 minutes, and then sonicate in a water bath for 15 minutes to obtain a clear suspension. The prepared sample was passed through a membrane (0.22 μm) three times to obtain a sample O-NP (1 mg / mL), which was stored at 4° C. for later use.

[0060] (2) Cell culture: Human colorectal cancer c...

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Abstract

The invention discloses a tumor microenvironment responsive surface charge reversible nano-drug delivery carrier which can be applied to specific detection and efficient treatment of MMP9 high-expression tumors. The functional amphiphilic molecular structure consists of three parts: oleic acid is a hydrophobic region of the molecule, the middle connecting part is MMP9 responsive broken polypeptide, and the other part is a polypeptide fragment rich in glutamic acid, so that a negatively charged amphiphilic molecule is formed. According to the nano delivery carrier, a basic framework with positive charges is composed of (2,3-Dioleoyloxy-propyl)-trimethylammonium-chloride, 1, 2-dioleoyl-sn-glycero-3-phosphocholine, soybean lecithin, DSPE-PEG2000; and after the basic framework is modified by functional amphiphilic molecules OMPE, surface charges are reversed into negative charges. Cell and animal experiments show that the nano delivery carrier can effectively deliver a detection probe anda tumor drug, and has very strong specificity detection and efficient treatment effects on MMP9 high-expression tumors.

Description

technical field [0001] The invention belongs to the field of nano-medicine, and in particular relates to a nano-medicine delivery carrier with reversible surface charge responsive to tumor microenvironment. Background technique [0002] Nanotechnology has attracted increasing interest and attention in cancer diagnosis and treatment, largely due to its unique and excellent properties, such as improving drug delivery efficiency, achieving long-term circulation in the body, and reducing toxicity. A large number of nano-drug carriers, such as liposomes, polymer micelles, dendrimers, and other nanotechnology modalities, have been used in clinical research and even approved for cancer treatment. Due to the leaky characteristics of tumor blood vessels, nanoparticles of suitable size can be preferentially enriched in tumor lesions, thereby effectively delivering chemotherapy drugs or imaging probes. However, there are multiple complex biological steps involved in the effective deli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/62A61K47/54A61K47/69A61K49/00C07K7/08B82Y5/00
CPCA61K49/0021A61K49/0052A61K49/0056A61K49/0084A61K47/542A61K47/62A61K47/6911B82Y5/00C07K7/08
Inventor 李鲁远张强哲韩秋菊
Owner NANKAI UNIV
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