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Human polypeptide for constructing tumor pH response micro-robot and application thereof

A micro-robot, human-derived technology, applied in the field of human-derived polypeptides, can solve serious problems, cancer cell toxicity, off-target effects, etc., achieve high biocompatibility, good drug side effects, and reduce drug side effects.

Active Publication Date: 2020-01-07
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it still remains a great challenge, for example, the development of new anti-angiogenic drugs, severe off-target effects, parallel energy supply of cancer cells and any toxicity and side effects

Method used

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  • Human polypeptide for constructing tumor pH response micro-robot and application thereof
  • Human polypeptide for constructing tumor pH response micro-robot and application thereof
  • Human polypeptide for constructing tumor pH response micro-robot and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Human-derived polypeptides are loaded with pirarubicin and indocyanine green through self-assembly function to form micro-robots, and the micro-robots are characterized:

[0033] Synthetic method: Firstly mix 1mg polypeptide freeze-dried powder and 1mg powdered indocyanine green thoroughly under the vortex shaker, and quickly centrifuge to collect the green mixture. Second, 5 μL of water was slowly added to the green mixture with constant stirring. After thorough mixing, 1 mL of water was added to dissolve the unbound polypeptide and indocyanine green, and the insoluble matter was collected by centrifugation at 5 000 rpm for 10 minutes. Finally, 1 mg of pirarubicin and 1 mL of water were added to the insoluble matter, shaken and mixed, and placed in an ice bath for 20 minutes of sonication.

[0034] Characterization: Using transmission electron microscopy and dynamic light scattering particle size analyzer (DLS) to study the morphology and particle size of t...

Embodiment 2

[0035] Example 2: Tumor acidic pH response of microrobots

[0036] Validation of release environment: acidic pH is the main feature of tumor tissue, so the acidic pH of tumors is often designed as a targeted trigger for drug release. Fluorescence spectroscopy detects changes in the emission intensity spectra of microrobots in the pH=7.4 and pH=6.5 environments. see Figure 4 , as the pH reached 6.5, we found that the solution containing the microrobots became turbid and green precipitates appeared; the fluorescence emission intensity of the solution was detected by fluorescence spectroscopy and found that the fluorescence intensity reversed, and the fluorescence intensity of indocyanine green decreased hundreds of times , the fluorescence intensity of pirarubicin increased dozens of times, which indicated that pirarubicin was released under acidic conditions, and the aggregation of indocyanine green caused fluorescence quenching.

Embodiment 3

[0037] Example 3: Anti-tumor growth of microrobots in an acidic pH environment

[0038] CCK-8 was used to detect the killing effect of microrobots on breast cancer cell MDA-231 in the environment of pH=7.4 and pH=6.5. 1×10 5Cells were seeded in 96-well plates, and after overnight growth, they were incubated with different concentrations of pirarubicin and micromachines for 2 hours in media with pH=7.4 and pH=6.5, respectively. After 2 hours, replace with fresh medium and continue to culture for another 24 hours, CCK-8 detects the viability of tumor cells. see Figure 6 : H, I, we found that microrobots and free indocyanine green have no inhibitory effect on the growth of cancer cells, indicating that they have strong biocompatibility; but with the increase of free pirarubicin, more and more of cancer cells were killed. When H+ triggered the microrobots to release pirarubicin, the cytotoxic effect of the microrobots was significantly enhanced, exceeding that of free pirarub...

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Abstract

The invention relates to a human polypeptide for constructing tumor pH response micro-robot and an application thereof. The amino acid sequence is as shown in SEQ ID NO.1. The micro-robot loads pirarubicin and indocyanine green through self-assembled human peptide nanoparticles, and combines chemotherapy, photothermal therapy and hunger therapy as well as real-time diagnosis of tumors. The micro-robot is triggered in the microenvironment of tumor acidic pH, releases pirarubicin and insoluble deposits and reverses their fluorescence emission intensity. The released pirarubicin selectively enters tumor nuclei and kills them, while insoluble deposits can accumulate in tumor blood capillaries, block oxygen and nutrition supply and realize cancer hunger treatment. Under the irradiation of nearinfrared laser, indocyanine green effectively accumulated in tumor can induce local hyperthermia. The reverse reversal of ph response of fluorescence emission intensity of micro-robot can be used forreal-time diagnosis of tumors in vivo.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a human-derived polypeptide for constructing a tumor pH-responsive microrobot and an application thereof. Background technique [0002] According to the latest statistics from the World Health Organization, an estimated 9.6 million people died of cancer in 2018. Early diagnosis and treatment of tumors is the most effective way to reduce cancer mortality. Due to its unique characteristics, nanotechnology has been widely concerned in the early diagnosis and treatment of cancer; and some nanomedicine materials have been approved for clinical cancer treatment. However, due to the suboptimal efficacy of nanotechnology in clinical trials, many promising nanotechnologies for cancer patients have not yet entered the clinic. Combination therapies, such as chemotherapy / immunotherapy, chemotherapy / photothermal therapy, chemotherapy / photodynamic therapy, and photodynamic / ...

Claims

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Application Information

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IPC IPC(8): C07K14/00A61K9/51A61K47/42A61K41/00A61K31/7048A61P35/00A61K49/00
CPCC07K14/001A61K9/5169A61K41/0052A61K31/7048A61P35/00A61K49/0093A61K2300/00
Inventor 吴国球蒋兴禄
Owner SOUTHEAST UNIV
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