CN110269942A - The targeting drug delivery system and synthetic method of a kind of photo-thermal in conjunction with chemotherapy - Google Patents
The targeting drug delivery system and synthetic method of a kind of photo-thermal in conjunction with chemotherapy Download PDFInfo
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Abstract
The invention belongs to field of medical technology, more particularly to a kind of targeting drug delivery system of photo-thermal in conjunction with chemotherapy, including target function molecule, chemotherapeutics and nano-carrier, the target function molecule is the antibody of active targeting, the chemotherapeutics is adriamycin, and the nano-carrier is the nano gold spherical of the Pegylation with photo-thermal effect；The chemotherapeutics is connected on the nano gold spherical of Pegylation by chemical bond and antibody.The utility model has the advantages that this drug-loading system has targeting, hypotoxicity, it is good by cellular uptake, with the property that photo-thermal effect can be reached to high efficiency anti-tumor in conjunction with chemotherapy, increase the aggregation at the H/N tumors tissue of oral cavity, damage of the chemotherapeutics to other normal tissues is reduced, the therapeutic effect of tumour is significantly improved.
Description
Technical field
A kind of targeting drug delivery system that the invention belongs to field of medical technology more particularly to photo-thermal in conjunction with chemotherapy and
Synthetic method.
Background technique
Traditional photo-thermal therapy is applied to deeper tumor tissues, because tumor tissues are deeper, offer limited effectiveness, and oral cavity neck is swollen
Tumor can have better effects because tumor tissues position table is shallow, but most delivery systems are poor to head and neck scale carcinoma targeting, not by light
Heat cure is combined together with chemotherapy.
Summary of the invention
The present invention is directed to above-mentioned problem, provides the targeting drug delivery system and synthesis of a kind of photo-thermal in conjunction with chemotherapy
Method.
According to an aspect of the invention, there is provided a kind of targeting drug delivery system of photo-thermal in conjunction with chemotherapy, including
Target function molecule, chemotherapeutics and nano-carrier, the target function molecule are the antibody of active targeting, the chemotherapeutic
Object is adriamycin, and the nano-carrier is the nano gold spherical of the Pegylation with photo-thermal effect；The chemotherapeutics passes through
Chemical bond and antibody are connected on the nano gold spherical of Pegylation.
Preferably, the antibody is podoplanin.
Preferably, used chemical bond is amido bond.
Preferably, the polyethylene glycol can be mono methoxy polyethylene glycol, or the poly- second containing other active groups
Glycol.
According to another aspect of the present invention, a kind of synthesis side of the targeting drug delivery of photo-thermal in conjunction with chemotherapy is provided
Method, comprising the following steps:
1) adriamycin is combined:
PEGylated AuNPs 1mL (2.46mg/mL) is taken, 15000rpm, 10min are centrifuged, removes supernatant, EDC/ is added
NHS solution rocks 2h, and after cleaning, the PBS solution of adriamycin (0.1mg/mL) is added, and rocks overnight, the carboxyl of the end PEG and Ah
The amino of mycin reacts to form DOX-AuNPs compound；
2) Podoplanin antibody is combined:
The addition of DOX-AuNPs compound is rocked into 2h for 37 DEG C of EDC/NHS solution, after cleaning, 40 μ L Podoplanin are added
Antibody, ultrasonic vibration 10s rock 7h, PBS cleaning, and the PBS solution that 5%BSA+0.05%Tween is added rocks synthesis overnight
Podoplanin-AuNPs-DOX compound.
Preferably, the EDC/NHS solution and the PBS solution are 37 DEG C.
Preferably, the concentration of the EDC/NHS solution is 80/20mg ratio, the capacity of addition is 2mL.
Compared with prior art, the advantages and positive effects of the present invention are,
1, overall structure of the present invention is simple, this drug-loading system has targeting, and hypotoxicity is good by cellular uptake,
With the property that photo-thermal effect can be reached to high efficiency anti-tumor in conjunction with chemotherapy.
2, the drug-loading system, which is applied to oral cavity H/N tumors, can have better effects because tumor tissues position table is shallow.
It 3, can be by active targeting (antibody) and passive on the basis of being combined together photo-thermal therapy and chemotherapy
Targeting (EPR (enhancing infiltration and delay) effect) increases the aggregation at the H/N tumors tissue of oral cavity, reduces chemotherapeutics
Damage to other normal tissues significantly improves the therapeutic effect of tumour.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, required use in being described below to embodiment
Attached drawing be briefly described, it should be apparent that, drawings in the following description are some embodiments of the invention, for ability
For the those of ordinary skill of domain, without any creative labor, it can also be obtained according to these attached drawings others
Attached drawing.
Fig. 1 is the synthetic method block diagram of targeting drug delivery system of the photo-thermal that provides of embodiment 1 in conjunction with chemotherapy；
Fig. 2 is the schematic diagram for the DOX burst size that embodiment 1 provides；
Fig. 3 is the schematic diagram that embodiment 2 provides；
Fig. 4 is the schematic diagram that embodiment 3 provides；
Fig. 5 is the schematic diagram that the DOX that embodiment 4 provides absorbs ability；
Fig. 6 is the schematic diagram that the Au that embodiment 4 provides absorbs ability；
Fig. 7 is the schematic diagram one that embodiment 5 provides；
Fig. 8 is the schematic diagram two that embodiment 5 provides；
Fig. 9 is the schematic diagram three that embodiment 5 provides；
Specific embodiment
To better understand the objects, features and advantages of the present invention, with reference to the accompanying drawings and examples
The present invention will be further described.It should be noted that in the absence of conflict, in embodiments herein and embodiment
Feature can be combined with each other.
In the following description, numerous specific details are set forth in order to facilitate a full understanding of the present invention, still, the present invention may be used also
To be implemented using other modes described herein are different from, therefore, the present invention is not limited to the specific of specification is described below
The limitation of embodiment.
According to an embodiment of the invention, providing the targeting drug delivery system and synthesis side of a kind of photo-thermal in conjunction with chemotherapy
Method, including target function molecule, chemotherapeutics and nano-carrier, the target function molecule are the antibody of active targeting, institute
Stating chemotherapeutics is adriamycin, and the nano-carrier is the nano gold spherical of the Pegylation with photo-thermal effect；The chemotherapy
Drug is connected on the nano gold spherical of Pegylation by chemical bond and antibody.
Further, the antibody is podoplanin.
Further, used chemical bond is amido bond.
Further, the polyethylene glycol can be mono methoxy polyethylene glycol, or the poly- second containing other active groups
Glycol.
Further, the targeted nano delivery system partial size is 100-130nm.
As shown in Figure 1, according to an embodiment of the invention, additionally providing a kind of targeting drug delivery of photo-thermal in conjunction with chemotherapy
Synthetic method, comprising the following steps:
S101, in conjunction with adriamycin:
PEGylated AuNPs 1mL (2.46mg/mL) is taken, 15000rpm, 10min are centrifuged, removes supernatant, EDC/ is added
NHS solution rocks 2h, and after cleaning, the PBS solution of adriamycin (0.1mg/mL) is added, and rocks overnight, the carboxyl of the end PEG and Ah
The amino of mycin reacts to form DOX-AuNPs compound；
S103, in conjunction with Podoplanin antibody:
The addition of DOX-AuNPs compound is rocked into 2h for 37 DEG C of EDC/NHS solution, after cleaning, 40 μ LPodoplanin are added
Antibody, ultrasonic vibration 10s rock 7h, PBS cleaning, and the PBS solution that 5%BSA+0.05%Tween is added rocks synthesis overnight
Podoplanin-AuNPs-DOX compound.
Further, the EDC/NHS solution and the PBS solution are 37 DEG C.
Further, the concentration of the EDC/NHS solution is 80/20mg ratio, and the capacity of addition is 2mL.
Specific embodiment: embodiment one: drug release detection
It chooses synthetic material Podoplanin-AuNPs-DOX to be dissolved in 1mLPBS in enclosed bag filter, bag filter is placed in
In 10mL medium, 37 DEG C, 120rpm rocks, and different time records the burst size of DOX (adriamycin).
The detection of two: 293T cytotoxicity of embodiment
The toxicity for selecting the nanogold (hereinafter referred to as AuNPs) of non-tumor cell 293T detection Pegylation, by 293T
Cell inoculation is added 5X103 in 96 orifice plates, every hole version.With the training of+1% mycillin culture medium of DMEM+10% fetal calf serum
Support for 24 hours, then suck culture medium, PBS is cleaned 3 times, be added containing various concentration (0,0.001,0.002,0.01,0.02,0.1,
0.2mg/mL, 200 holes μ L/) AuNPs culture medium culture 48h, then with MTT kit test cell activity (cell Proliferation with
And citotoxicity detection kit).AuNPs is obtained after handling cell 48h, cell viability is still very high, it was demonstrated that AuNPs has low
Toxicity.
Embodiment three: antitumous effect detection
Tumour cell Cal27 is selected to detect Free DOX, AuNPs+PTT, Podplanin-AuNPs-DOX,
The antitumous effect of Podoplanin-AuNPs-DOX+PTT.Cal27 cell inoculation is added in 96 orifice plates, every orifice plate
5X103 cell.For 24 hours with+1% mycillin culture medium culture of DMEM+10% fetal calf serum, culture medium is then sucked, PBS is clear
It washes 3 times, (0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8 μM) Free DOX (experimental group 1) containing various concentration is added,
AuNPs (experimental group 2), Podoplanin-AuNPs-DOX (experimental group 3), Podoplanin-AuNPs-DOX's (experimental group 4)
Culture medium culture 48h is (experimental group 2 and experimental group 4 when the culture medium culture containing material is added for 24 hours, with the 532nm of 1W/cm2
Laser irradiates 15min respectively), then with MTT kit test cell activity.Know that Podoplanin-AuNPs-DOX system can
With by chemotherapy with photo-thermal therapy is good combines, antitumous effect with higher.
Example IV: cellular uptake ability detection
(1) DOX absorbs ability
Tumour cell Cal27 is selected to detect intake ability.1.5X is added in 24 orifice plates, every hole in Cal27 cell inoculation
105 cells.When cell reaches 70%, the culture of Podoplanin-AuNPs-DOX (the 2 μ g/mL of DOX of equivalent) is added
Base sucks culture medium after cultivating 0.5h, and PBS is washed 3 times, and fluorescence inverted microscope observation is ingested situation.
(2) Au absorbs ability
Tumour cell Cal27 is selected to detect intake ability.2X will be added in 12 orifice plates, every hole in Cal27 cell inoculation
105 cell pellet overnights are added and contain AuNPs (40 μ g/mL) respectively, Podoplanin-AuNPs-DOX's (AuNPs of equivalent)
Culture medium cultivates 0.5h, 1h, 2h, 6h respectively, sucks culture medium, and PBS is cleaned three times, and pancreatin is added and is digested, is added and contains blood
Clear DMEM terminates digestion, is centrifuged 3min, 1000rpm, counts, and ICP-MS detects intracellular Au content.
Embodiment five: internal antitumous effect detection
4 week old nude mice 30 is chosen, Cal27 tumour cell (2X105,100 μ L PBS) are subcutaneously injected in left hind, growth 3
Zhou Hou is arbitrarily divided into 5 groups (n=6), is grouped as follows (A) physiological saline (B) Free DOX (2mg/kg) (C) AuNPs+ illumination (D)
Podoplanin-AuNPs-DOX (the DOX 2mg/kg of equivalent) (E) Podoplanin-AuNPs-DOX illumination (DOX of equivalent
2mg/kg), an every group of mouse co-injection of tail vein injection 3 times every three days, after light group per injection for 24 hours, 532nm laser
15min, 48h after last time is injected are irradiated, mouse is put to death, and takes tumour, and the heart, liver, spleen, lung, kidney, weighing, light group mouse is swollen
Tumor sends to the content for being Au in ICP-MS test tumour.AuNPs+ light group and Podoplanin-AuNPs-DOX+ light group tool
Have a good ability for converting light energy into thermal energy, temperature can rise to 42 DEG C at 6 minutes or so, and physiological saline group and
DOX group does not change substantially.Confirm that Podoplanin-AuNPs-DOX system can be good by chemotherapy and photo-thermal therapy in vivo
Be combined together, have preferable antitumous effect.Podoplanin-AuNPs-DOX group antitumous effect is 70%,
Podoplanin-AuNPs-DOX+ light group antitumous effect can reach 82.6%.
The above described is only a preferred embodiment of the present invention, being not that the invention has other forms of limitations, appoint
What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc.
It imitates embodiment and is applied to other fields, but without departing from the technical solutions of the present invention, according to the technical essence of the invention
Any simple modification, equivalent variations and remodeling to the above embodiments, still fall within the protection scope of technical solution of the present invention.
Claims (8)
1. a kind of targeting drug delivery system of photo-thermal in conjunction with chemotherapy, which is characterized in that including target function molecule, chemotherapeutic
Object and nano-carrier, the target function molecule are the antibody of active targeting, and the chemotherapeutics is adriamycin, the nanometer
Carrier is the nano gold spherical of the Pegylation with photo-thermal effect；The chemotherapeutics is connected to poly- by chemical bond and antibody
On the nano gold spherical of glycation.
2. a kind of targeting drug delivery system of the photo-thermal according to claim 1 in conjunction with chemotherapy, which is characterized in that described
Antibody is podoplanin.
3. a kind of targeting drug delivery system of the photo-thermal according to claim 1 in conjunction with chemotherapy, which is characterized in that adopted
Chemical bond is amido bond.
4. a kind of targeting drug delivery system of the photo-thermal according to claim 1 in conjunction with chemotherapy, which is characterized in that described
Polyethylene glycol can be mono methoxy polyethylene glycol, or the polyethylene glycol containing other active groups.
5. a kind of targeting drug delivery system of the photo-thermal according to claim 1 in conjunction with chemotherapy, which is characterized in that use
Targeted nano delivery system partial size be 100-130nm.
6. a kind of synthetic method of targeting drug delivery system of photo-thermal in conjunction with chemotherapy, which is characterized in that be used for claim 1
The preparation of targeting drug delivery of the photo-thermal in conjunction with chemotherapy, comprising the following steps:
1) adriamycin is combined:
PEGylated AuNPs 1mL (2.46mg/mL) is taken, centrifugation 15000rpm, 10min are carried out, removes supernatant, and be added
EDC/NHS solution rocks 2h, and after cleaning, the PBS solution of adriamycin (0.1mg/mL) is added, and rocks overnight, the carboxyl of the end PEG
It reacts to form DOX-AuNPs compound with the amino of adriamycin；
2) Podoplanin antibody is combined:
The addition of DOX-AuNPs compound is rocked into 2h for 37 DEG C of EDC/NHS solution, after cleaning, it is anti-that 40 μ L Podoplanin are added
Body, ultrasonic vibration 10s, and 7h is rocked, after being cleaned by PBS, the PBS solution that 5%BSA+0.05%Tween is added was rocked
Night synthesizes Podoplanin-AuNPs-DOX compound.
7. a kind of synthetic method of targeting drug delivery system of the photo-thermal according to claim 6 in conjunction with chemotherapy, special
Sign is that the EDC/NHS solution and the PBS solution are 37 DEG C.
8. a kind of synthetic method of targeting drug delivery system of the photo-thermal according to claim 6 in conjunction with chemotherapy, special
Sign is that the concentration of the EDC/NHS solution is 80/20mg ratio, and the capacity of addition is 2mL.
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|---|---|---|---|---|
| CN113058036A (en) * | 2021-04-19 | 2021-07-02 | 上海交通大学医学院附属第九人民医院 | SCGB3A2-PDPN-RhoA signal channel as drug target for inhibiting lung inflammation factor storm |
| CN113058036B (en) * | 2021-04-19 | 2022-09-13 | 上海交通大学医学院附属第九人民医院 | SCGB3A2-PDPN-RhoA signal channel as drug target for inhibiting lung inflammatory factor storm |
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