CN110292634A - A kind of photo-thermal chemotherapy combined therapeutic reagent and its preparation method and application - Google Patents
A kind of photo-thermal chemotherapy combined therapeutic reagent and its preparation method and application Download PDFInfo
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- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
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- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6923—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
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Abstract
The invention discloses a kind of photo-thermal chemotherapy combined therapeutic reagent and its preparation method and application, the preparation method is comprising steps of (1) prepares black phosphorus nanometer sheet；(2) graphene quantum dot of 2nd area of near-infrared response is prepared；And the graphene quantum dot that black phosphorus nanometer sheet and 2nd area of near-infrared respond is placed in phosphate buffered saline solution at room temperature and is vigorously stirred for 24 hours by (3), the surface of black phosphorus nanometer sheet is deposited on the graphene quantum dot for responding infrared 2nd area, through centrifugal treating to get photo-thermal chemotherapy combined therapeutic reagent.The preparation method of photo-thermal chemotherapy combined therapeutic reagent provided by the invention overcomes black phosphorus nanometer sheet disadvantage easy to oxidize in air by the way that functionalization graphene quantum dot is coated on black phosphorus surface；And the photo-thermal chemotherapy combined therapeutic reagent prepared by it has high photothermal conversion efficiency, excellent tumour cell targeting and biocompatibility.
Description
Technical field
The present invention relates to Nano medication technical field more particularly to a kind of photo-thermal chemotherapy combined therapeutic reagent and its systems
Preparation Method and application.
Background technique
Cancer in the past few decades in be still threaten human health major disease.Global annual about 14,000,000
New cancer patient has 8,000,000 people to die of cancer-related diseases every year at the same time.Photo-thermal chemotherapy combined therapy is as conventional cancer
The promising substituted or supplemented therapy of one kind for the treatment of technology (operation, radiation and chemotherapy), it is unspent same in systemic chemotherapy
When, drug quick release caused by the synergistic effect and thermotherapy of photo-thermal therapy method itself, to the inhibition of Local tumor growth have compared with
High curative effect, and can be avoided the damage of normal tissue.As a kind of effective noninvasive laser therapy method, photo-thermal chemotherapy
Conjoint therapy can treat a plurality of types of cancers.
Ideal photo-thermal chemotherapy combined therapeutic reagent should have high photothermal conversion efficiency, and have in tumour good
Accumulation.Wherein graphene quantum dot can be with active targeting swollen as a kind of photo-thermal chemotherapy combined therapeutic reagent carrier medicament
It accumulates, and a variety of imaging modes and treatment function can be integrated on a platform in tumor.Graphene quantum dot is a kind of
The emerging zero dimension material with biocompatibility, the sulfonic acid functional of its marginal position can be such that it is dispersed stably in water,
Even if still keeping high fluorescence activity after being annealed to 250 DEG C, it does not have cytotoxicity and has environment friendly, to graphene amount
Sub- point edge, which is chemically modified, can effectively facilitate healthy cell proliferation and differentiation.
The two-dimensional layers nano material such as black phosphorus, transition metal carbide or carbonitride, since it is imitated with excellent photo-thermal
It answers, makes it that there is potential application prospect in photo-thermal chemotherapy combined therapeutic reagent and drug delivery.Black phosphorus is as a kind of novel
Two-dimensional material, specific surface area with higher, excellent electrochemical properties and good surface property, this is examined for it in Raman
The application in many fields such as survey, biomedicine, chemical sensitisation has established solid foundation compared to other two dimensions such as graphene
Material, metastable physicochemical properties of black phosphorus and to the highly selective of moieties allow it to be provided with very in terms of sense
Big advantage is however, black phosphorus nanometer sheet is easy to oxidize in air, and generation is adsorbed on the insulating oxide phosphorus on black phosphorus surface, seriously
Hinder application of the black phosphorus in drug.
Currently, functionalization graphene quantum dot is as light thermit powder it has been reported that still functionalization graphene quantum dot coats
Black phosphorus, as photo-thermal chemotherapy combined therapeutic reagent, there is not been reported.
Summary of the invention
The present invention is to solve the above problem in the prior art, propose a kind of photo-thermal chemotherapy combined therapeutic reagent and its
Preparation method and application.
To achieve the above object, the invention adopts the following technical scheme:
The first aspect of the invention is to provide a kind of preparation method of photo-thermal chemotherapy combined therapeutic reagent, including such as
Lower step:
(1) black phosphorus nanometer sheet is prepared
S11, appropriate black phosphorus powder is added in n-methyl-2-pyrrolidone solution, is carried out fully dispersed；
S12, dispersion liquid is subjected to centrifugal treating, removes remaining unstripped particle, and remove extra N- methyl-
2-Pyrrolidone, the precipitating after separation are black phosphorus nanometer sheet；
S13, the black phosphorus nanometer sheet after separation is dissolved in ethyl alcohol, it is spare；
(2) graphene quantum dot of 2nd area of near-infrared response is prepared
S21, suitable 1,3,6- trinitrotoluene, polyethyleneimine are separately added into water, are uniformly mixed；
S22, mixed suspension is heated at a temperature of 220-250 DEG C 5min, be cooled to room temperature；
S23, the insoluble sludge in mixed liquor is removed using filtering with microporous membrane；
S24, the bag filter for being 3500Da with molecular weight by filtrate dialyse two days, obtain the graphite of 2nd area of near-infrared response
Alkene quantum dot, it is spare；
(3) graphene quantum dot/black phosphorus nanometer reagent of 2nd area of near-infrared response is prepared
S31, the graphene amount for responding 2nd area of near-infrared of the black phosphorus nanometer sheet of step (1) preparation and step (2) preparation
Son point, which is placed at room temperature in phosphate buffered saline solution (PBS, pH=8.0), to be vigorously stirred for 24 hours, the stone that infrared 2nd area is responded
Black alkene quantum dot is deposited on the surface of black phosphorus nanometer sheet；
S32, mixed liquor is subjected to centrifugal treating, removes unbonded graphene quantum dot to get graphene quantum dot/black
Phosphorus nanometer reagent.
Further, in step S11, the molar ratio of the black phosphorus powder and the n-methyl-2-pyrrolidone solution is
3.5:1。
Further, in step S11, the dispersing technology are as follows: by the mixing of black phosphorus powder and n-methyl-2-pyrrolidone
Liquid is placed in the Ultrasound Instrument that ultrasonic power is 300 watts, and the sonic oscillation 10h under condition of ice bath.
Further, it in step S21, is first centrifuged 8 minutes with 4000 revs/min, to remove remaining unstripped particle；Again
Supernatant is centrifuged 5 minutes with 4000 revs/min, to remove extra n-methyl-2-pyrrolidone.
Further, in step 21, described 1, the molar ratio of 3,6- trinitrotoluenes and the polyethyleneimine is 5:3；
The dosage of the water is 10mL.
Further, in step 22, the suspension is transferred in microwave reactor, heats 5 at a temperature of 230 DEG C
Minute, then naturally cool to room temperature.
Further, in step 23, the aperture of the miillpore filter is 0.15-0.3mm.
Further, in step 31, the graphene quantum dot of the infrared 2nd areas response and the matter of the black phosphorus nanometer sheet
Amount is than being 5:1.
Further, in step 31, the graphene quantum dot of the infrared 2nd areas response is in the phosphate buffered saline solution
In concentration be 500 μ g/mL；Concentration of the black phosphorus nanometer sheet in the phosphate buffered saline solution is 100 μ g/mL.
The second aspect of the invention is to provide a kind of photo-thermal chemotherapy combined treatment of method preparation as described above
Reagent.
The third aspect of the invention is to provide one kind photo-thermal chemotherapy combined therapeutic reagent described above and activity
Biomolecule is mutually compounded in the application prepared in anticancer drug.
The present invention by adopting the above technical scheme, compared with prior art, has the following technical effect that
(1) preparation method of photo-thermal chemotherapy combined therapeutic reagent provided by, by by functionalization graphene quantum
Point is coated on black phosphorus surface, overcomes black phosphorus nanometer sheet disadvantage easy to oxidize in air；
(2) graphene quantum dot prepared by it/black phosphorus reagent has high photothermal conversion efficiency, excellent tumour cell
Targeting and biocompatibility；
(3) use nanometer biotechnology by the bioactive molecules such as anti-tumor drug (API) and photo-thermal chemotherapy combined
Therapeutic reagent is mutually compound, and then changes the property of pharmacokinetics, drug effect and pharmacology of contained API etc. using nano effect
Matter, and the Nanoscale assemblies of significant clinical advantage are obtained, and effectively treat a plurality of types of cancers in conjunction with photo-thermal chemotherapy combined therapy
Disease.
Detailed description of the invention
A) TEM of black phosphorus nanometer sheet schemes in Fig. 1；B) the TEM figure of single black phosphorus nanometer sheet；C) HRTEM of black phosphorus nanometer sheet
Figure；D) graphene quantum dot/black phosphorus nanometer reagent TEM schemes；E) graphene quantum dot/black phosphorus nanometer reagent HRTEM schemes；f)
Graphene quantum dot/black phosphorus nanometer reagent AFM figure；
A) the X ray diffracting spectrum of black phosphorus nanometer sheet in Fig. 2；B) graphene quantum dot/black phosphorus nanometer reagent X-ray is spread out
Penetrate map；
Fig. 3 is black phosphorus nanometer sheet and graphene quantum dot/black phosphorus nanometer reagent infrared spectroscopy；
Fig. 4 is black phosphorus nanometer sheet and graphene quantum dot/black phosphorus nanometer reagent Raman spectrum；
A) the XPS score of black phosphorus nanometer sheet in Fig. 5；B) graphene quantum dot/black phosphorus nanometer reagent XPS score；C) stone
Black alkene quantum dot/black phosphorus nanometer reagent high-resolution N 1s spectrogram；
A) the fluorescence spectrum of graphene quantum dot in Fig. 6；B) graphene quantum dot and graphene quantum dot/black phosphorus nanometer examination
The fluorescence emission spectrum of agent；
Fig. 7 be under same concentration graphene quantum dot, black phosphorus nanometer sheet, graphene quantum dot/black phosphorus nanometer reagent can
The comparison that light-exposed and near infrared region absorbs；
Fig. 8 is the graphene quantum dot/black phosphorus nanometer examination under conditions of different 1064nm laser powers is irradiated 5 minutes
The infrared thermal imaging figure of agent；
Fig. 9 is graphene quantum dot/black phosphorus nanometer reagent in 0.6W cm-2To Hela cell survival under the conditions of illumination 5 minutes
The influence of rate；
Figure 10 is injected intravenously physiological saline, free adriamycin and graphene quantum dot/black respectively during being oncotherapy
Under conditions of phosphorus/adriamycin composite nanometer particle, the change curve of corresponding gross tumor volume.
Specific embodiment
The present invention is described in more detail below by specific embodiment, for a better understanding of the present invention,
But following embodiments are not intended to limit the scope of the invention.
Embodiment 1
A kind of preparation method of photo-thermal chemotherapy combined therapeutic reagent, specifically comprises the following steps:
(1) black phosphorus nanometer sheet is prepared
S11,25-50mg black phosphorus powder is added in the beaker of the n-methyl-2-pyrrolidone solution equipped with 25-50mL,
Ice water is placed in the Ultrasound Instrument that ultrasonic power is 300 watts, sonic oscillation 8-10h, progress are abundant under condition of ice bath beaker
Dispersion；
S12, dispersion liquid is subjected to centrifugal treating, is centrifuged 8 minutes with 4000 revs/min to remove remaining unstripped
Grain；Supernatant is centrifuged 5 minutes with 4000 revs/min, to remove n-methyl-2-pyrrolidone, the precipitating after separation is black phosphorus
Nanometer sheet；
S13, the black phosphorus nanometer sheet after separation is dissolved in ethyl alcohol, is saved in ethanol to prevent black phosphorus nanometer sheet from aoxidizing,
It is spare；
(2) graphene quantum dot of 2nd area of near-infrared response is prepared
S21, the polyethyleneimine of the 1,3,6-trinitrotoluenes of 0.025mg, 0.4mg are separately added into the water containing 10mL
Beaker in, by sonic oscillation, be uniformly mixed；
S22, mixed suspension is transferred in microwave reactor, 230 DEG C at a temperature of heat 5 minutes, then
Naturally cool to room temperature；
S23, using the aperture 0.22mm filtering with microporous membrane removal mixed liquor in insoluble sludge；
S24, the bag filter for being 3500Da with molecular weight by filtrate dialyse two days, obtain the graphite of 2nd area of near-infrared response
Alkene quantum dot, it is spare；
(3) graphene quantum dot/black phosphorus nanometer reagent of 2nd area of near-infrared response is prepared
S31, the graphene amount for responding 2nd area of near-infrared of the black phosphorus nanometer sheet of step (1) preparation and step (2) preparation
Son point, which is placed at room temperature in phosphate buffered saline solution (PBS, pH=8.0), to be vigorously stirred for 24 hours, the stone that infrared 2nd area is responded
Black alkene quantum dot (500 μ g mL-1) it is deposited on black phosphorus nanometer sheet (100 μ gmL-1) surface；
S32, mixed liquor is centrifuged 5 minutes by 10000 revs/min, removes unbonded graphene quantum dot to get graphite
Alkene quantum dot/black phosphorus nanometer reagent.
As shown in Figure 1, wherein a) TEM of black phosphorus nanometer sheet schemes；B) the TEM figure of single black phosphorus nanometer sheet；C) black phosphorus nanometer
The HRTEM of piece schemes；D) graphene quantum dot/black phosphorus nanometer reagent TEM schemes；E) graphene quantum dot/black phosphorus nanometer reagent
HRTEM figure；F) graphene quantum dot/black phosphorus nanometer reagent AFM schemes；Analysis is it is found that the black phosphorus prepared using the method for the present invention
The average grain diameter of nanometer sheet is 100~200nm, good dispersion.Black squama can be observed from the TEM figure of single black phosphorus nanometer sheet to receive
The individual layer laminated structure of rice piece, high power TEM show that it has apparent lattice fringe, and spacing of lattice is 0.29nm.Average thickness
For 2.0nm.After graphene quantum dot load, the spacing of lattice of graphene quantum dot can be clearly seen in black phosphorus nanometer sheet
For 0.21nm.
As shown in Fig. 2, a) the X ray diffracting spectrum of black phosphorus nanometer sheet；B) graphene quantum dot/black phosphorus nanometer reagent X
X ray diffraction map；Analysis is it is found that X ray diffracting spectrum shows that graphene quantum dot is successfully supported in black phosphorus nanometer sheet.
As shown in figure 3, being black phosphorus nanometer sheet and graphene quantum dot/black phosphorus nanometer reagent infrared spectroscopy；Analysis it is found that
Infrared spectroscopy shows black phosphorus nanometer sheet in 1000,1200 and 1620cm-1, and there are P-O, P-P-O and P=O stretching vibration in place, load
After complete quantum dot, there are 1362 (C-H), 1435 (C-N) and 1662cm-1(N-H), it was demonstrated that graphene quantum dot successfully loads
In black phosphorus nanometer sheet.
As shown in figure 4, being black phosphorus nanometer sheet and graphene quantum dot/black phosphorus nanometer reagent Raman spectrum；Analysis it is found that
The Raman figure of black phosphorus nanometer sheet shows A1g, B2g, the peak Ag2, A1g in graphene quantum dot/black phosphorus reagent, B2g, Ag2 peak value point
Other red shift 2.1,3.2,3.1cm-1。
As shown in figure 5, wherein a) the XPS score of black phosphorus nanometer sheet；B) graphene quantum dot/black phosphorus nanometer reagent XPS
Score；C) graphene quantum dot/black phosphorus nanometer reagent high-resolution N 1s spectrogram；It is found that graphene quantum dot/black phosphorus nanometer examination
Agent has strong C 1s, N 1s, O 1s and P 2p signal, it was demonstrated that black phosphorus nanometer reagent is loaded successfully into black phosphorus nanometer sheet.
As shown in fig. 6, the wherein a) fluorescence spectrum of graphene quantum dot；B) graphene quantum dot and graphene quantum dot/
The fluorescence emission spectrum of black phosphorus nanometer reagent；Analysis is best to send out it is found that a length of 415nm of the optimum excitation wave of graphene quantum dot
The a length of 475nm of ejected wave.
And it as shown in fig. 7, is received for graphene quantum dot, black phosphorus nanometer sheet, graphene quantum dot/black phosphorus under same concentration
The comparison that rice reagent is absorbed in visible light and near infrared region；Analysis is it is found that graphene quantum dot/black phosphorus nanometer examination under same concentrations
Agent near infrared absorption is most strong, and simple graphene quantum dot takes second place, and black phosphorus nanometer sheet is most weak.
Embodiment 2
The external Photothermal characterisation of graphene quantum dot/black phosphorus nanometer reagent of infrared 2nd areas response is investigated
By graphene quantum dot/black phosphorus nanometer reagent dilutions at different concentration (0-600 μ g mL-1), it is then transferred to
The centrifuge tube of 1.5mL, with the laser of 1064nm with 0.6W cm-2Power density irradiate 5 minutes, every 20s with it is infrared it is hot at
As the variation of instrument record temperature.Change power (0.4-0.8W cm-2) use 1064nm 600 μ g mL of laser illumination-1Graphene
Quantum dot/black phosphorus nanometer reagent is to measure influence of the different capacity to heating.It is 0.6W cm by power density-21064nm
Graphene quantum dot/black phosphorus nanometer reagent photostability is probed into the variation of temperature under laser reirradiation 5min.
As shown in figure 8, under conditions of different 1064nm laser powers is irradiated 5 minutes, graphene quantum dot/black phosphorus
The infrared thermal imaging figure of nanometer reagent.It analyzes it is found that graphene quantum dot/black phosphorus nanometer reagent solution is in 0.6W cm-2Power
Under irradiation, 24.5 DEG C are risen in 5 minutes.
Embodiment 3
The external photo-thermal therapy of graphene quantum dot/black phosphorus nanometer reagent:
The present invention provides on by mtt assay to by graphene quantum dot/black phosphorus nanometer reagent treated cell into
The detection of row cell viability.By human cervical carcinoma cell (Hela) kind into 96 orifice plates, the density in every hole is 5000, and culture 24 is small
When, graphene quantum dot/black phosphorus nanometer reagent of various concentration is then added, after culture 4 hours, with 0.6W cm-21064nm
It laser illumination 5 minutes, cell is then continued culture reaches within 24 or 48 hours that the MTT of 20 μ L is added after the time to be measured, then
Continuation continues to be incubated for 4 hours at 37 DEG C, and finally culture solution is sucked out, and the dimethyl sulfoxide that 150 μ L are added in every hole is dissolved,
The absorption at 490nm is measured with microplate reader.
As shown in figure 9, being graphene quantum dot/black phosphorus nanometer reagent in 0.6W cm-2To Hela under the conditions of illumination 5 minutes
The influence of cell survival rate；Analysis is it is found that graphene quantum dot/black phosphorus/adriamycin composite nanometer particle under laser irradiation condition
Show apparent cytotoxicity.
Embodiment 4
The external combination therapy Performance Evaluation of graphene quantum dot/black phosphorus/adriamycin composite nanometer particle:
Cell is carried out to through graphene quantum dot/black phosphorus/adriamycin composite nanometer particle treated cell by mtt assay
The detection of activity, by Hela cell kind into 96 orifice plates, the density in every hole is 5000, after culture 4 hours, with 0.4W cm-2's
1064nm laser illumination 5 minutes, then, cell is incubated for 24 hours or 48 hours again, it is living to measure cell to carry out MTT measurement
Power.
Embodiment 5
Combination therapy Performance Evaluation in graphene quantum dot/black phosphorus/adriamycin composite nanometer particle body:
By the 4T1 cell subcutaneous inoculation of 100 μ L (1,000,000) at 3-5 weeks Female nude mice armpit, band gross tumor volume is grown to
When 100-200mm3, nude mice is divided into 7 groups (every group 5): (1) the free adriamycin in physiological saline, (2), (3) black phosphorus nanometer
Piece, (4) physiological saline+illumination (0.6W cm-2, 1064nm), (5) graphene quantum dot/black phosphorus+illumination (0.2W cm-2、
808nm), (6) graphene quantum dot/black phosphorus+illumination (0.6W cm-2, 1064nm), (7) graphene quantum dot/black phosphorus/adriamycin
(0.4W cm-2,1064nm).By the way that in intravenous injection (200 μ L) to tumor-bearing mice body, the volume for every other day measuring tumour is big
The weight of small daily record nude mice.
As shown in Figure 10, be the variation of gross tumor volume during oncotherapy, by intravenous injection physiological saline, free Ah
Mycin and graphene quantum dot/black phosphorus/adriamycin composite nanometer particle are 0.4Wcm in power density-2Under conditions of use
1064nm laser irradiation 5min.Gross tumor volume every other day measures once.Analysis is it is found that intravenous injection graphene quantum dot/black
After phosphorus/adriamycin nano reagent, in 0.4W cm-2It is irradiated 5 minutes under power, gross tumor volume reduces until disappearing.
Specific embodiments of the present invention are described in detail above, but it is merely an example, the present invention is simultaneously unlimited
It is formed on particular embodiments described above.To those skilled in the art, any couple of present invention carries out equivalent modifications and
Substitution is also all among scope of the invention.Therefore, without departing from the spirit and scope of the invention made by equal transformation and
Modification, all should be contained within the scope of the invention.
Claims (10)
1. a kind of preparation method of photo-thermal chemotherapy combined therapeutic reagent, which comprises the steps of:
(1) black phosphorus nanometer sheet is prepared
S11, appropriate black phosphorus powder is added in n-methyl-2-pyrrolidone solution, is carried out fully dispersed；
S12, dispersion liquid is subjected to centrifugal treating, removes remaining unstripped particle, and remove extra N- methyl -2- pyrrole
Pyrrolidone, the precipitating after separation are black phosphorus nanometer sheet；
S13, the black phosphorus nanometer sheet after separation is dissolved in ethyl alcohol, it is spare；
(2) graphene quantum dot of 2nd area of near-infrared response is prepared
S21, suitable 1,3,6- trinitrotoluene, polyethyleneimine are separately added into water, are uniformly mixed；
S22, mixed suspension is heated at a temperature of 220-250 DEG C 5min, be cooled to room temperature；
S23, the insoluble sludge in mixed liquor is removed using filtering with microporous membrane；
S24, the bag filter for being 3500 Da by filtrate molecular weight, dialyse two days, obtain the graphene amount of 2nd area of near-infrared response
It is sub-, it is spare；
(3) graphene quantum dot/black phosphorus nanometer reagent of 2nd area of near-infrared response is prepared
S31, the graphene quantum dot for responding 2nd area of near-infrared of the black phosphorus nanometer sheet of step (1) preparation and step (2) preparation
It is placed in phosphate buffered saline solution (PBS, pH=8.0) and is vigorously stirred for 24 hours at room temperature, the graphene that infrared 2nd area is responded
Quantum dot is deposited on the surface of black phosphorus nanometer sheet；
S32, mixed liquor is subjected to centrifugal treating, removes unbonded graphene quantum dot and is received to get graphene quantum dot/black phosphorus
Rice reagent.
2. the preparation method of photo-thermal chemotherapy combined therapeutic reagent according to claim 1, which is characterized in that step
The molar ratio of black phosphorus powder described in S11 and the n-methyl-2-pyrrolidone solution is 3.5:1.
3. the preparation method of photo-thermal chemotherapy combined therapeutic reagent according to claim 1, which is characterized in that step
In S11, the dispersing technology are as follows: it is 300 watts that the mixed liquor of black phosphorus powder and n-methyl-2-pyrrolidone, which is placed in ultrasonic power,
Ultrasound Instrument in, and the sonic oscillation 10h under condition of ice bath.
4. the preparation method of photo-thermal chemotherapy combined therapeutic reagent according to claim 1, which is characterized in that step
It is first centrifuged 8 minutes with 4000 revs/min in S21, to remove remaining unstripped particle；Again by supernatant with 4000 revs/min from
The heart 5 minutes, to remove extra n-methyl-2-pyrrolidone.
5. the preparation method of photo-thermal chemotherapy combined therapeutic reagent according to claim 1, which is characterized in that step 21
Described in the molar ratio of 1,3,6- trinitrotoluene and the polyethyleneimine be 5:3.
6. the preparation method of photo-thermal chemotherapy combined therapeutic reagent according to claim 1, which is characterized in that step 23
Described in miillpore filter aperture be 0.15-0.3mm.
7. the preparation method of photo-thermal chemotherapy combined therapeutic reagent according to claim 1, which is characterized in that step 31
Described in the graphene quantum dot of infrared 2nd area response and the mass ratio of the black phosphorus nanometer sheet be 5:1.
8. the preparation method of photo-thermal chemotherapy combined therapeutic reagent according to claim 1, which is characterized in that step 31
Described in concentration of the graphene quantum dot in the phosphate buffered saline solution of infrared 2nd area response be 500 μ g/mL；It is described black
Concentration of the phosphorus nanometer sheet in the phosphate buffered saline solution is 100 μ g/mL.
9. a kind of photo-thermal chemotherapy combined therapeutic reagent prepared such as any one of claim 1-8 the method.
10. a kind of photo-thermal chemotherapy combined therapeutic reagent as claimed in claim 9 is mutually compounded in system with active biomolecule
Application in standby anticancer drug.
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Citations (5)
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| CN108772079A (en) * | 2018-04-26 | 2018-11-09 | 昆明理工大学 | A kind of preparation method of nanometer of black phosphorus/graphene composite material |
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