CN110279858A - A kind of boracic preparation and its preparation method and application having cell nucleus targeting - Google Patents
A kind of boracic preparation and its preparation method and application having cell nucleus targeting Download PDFInfo
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Abstract
The invention discloses a kind of boracic preparation and its preparation method and application for having cell nucleus targeting, the boracic preparation is made of 1- bromomethyl neighbour carborane and the cell nucleus targeting drug containing active amino；Alternatively, being made of 1- bromomethyl neighbour carborane, 2 generation dendroid polyamide and hyaluronic acid；Alternatively, being made of 4- boron-L-phenylalanine, di-tert-butyl dicarbonate and 2 generation dendroid polyamide；Alternatively, being made of adjacent carborane and dendroid polyethyleneimine.The boracic preparation for having cell nucleus targeting of the invention, while showing preferable tumour cell intake, moreover it is possible to which further targeting is positioned at neoplastic cell nuclei, is the Novel boron trapping agent for having cell nucleus targeting.
Description
Technical field
The present invention relates to biomedicine technical fields, and in particular to a kind of boracic preparation for having cell nucleus targeting and its
Preparation method and application.
Background technique
Tumour especially malignant tumour is to seriously threaten the principal disease of human health, all to patient, family and society
Bring heavy burden.In recent years, tumor incidence is in rising trend, the prediction of international cancer tumor center, the World Health Organization
The year two thousand twenty whole world is annual newly-increased to suffer from cancer occurrence numbers and is up to 15,000,000.At present the essential therapeutic arsenals of tumour have operation, radiotherapy,
Chemotherapy, immunization therapy etc., but quite a few tumor patient (such as high-level tumour, recurrent tumor, metastatic tumo(u)r) is not
Can therefrom it benefit, the new tool for exploring oncotherapy is of far-reaching significance.
Boron neutron capture therapy (boron neutron capture therapy, BNCT) be it is a kind of based on nuclear capture and
The novel radiotherapy mode of fission reaction, it destroys cancer cell by the way that the nuclear reaction in tumour cell occurs.When
Stable isotope10Boron (10B after) targeting is transported to tumour cell, using the low energy neutron (heat to tissue cell insult very little
Neutron, epithermal neutron) irradiation excitation10Fission reaction occurs for B:10B+n→4He(1.47MeV)+7Li(0.84MeV)+γ
(0.48MeV)。4He and7Li is High Linear energy transmission ray, has stronger biology than the X-ray that conventional tumor radiotherapy uses
Learn effect.4He and7The range of Li in the tissue is only 5~9 μm, and hazard boundary is confined to high intake10The tumor tissue cell of B,
And to not absorbing10The normal tissue of B is almost without injury.Therefore, BNCT alternative killing tumor cell in cell dimensions,
It reduces the toxic side effect to patient as far as possible simultaneously, has good cancer therapeutic applications prospect.
Successful BNCT needs the boron capture agent to tumour high-affinity, mainly has two aspects to require: (1) tumor tissues phase
To height intake, (every gram of tumor tissues at least absorb 20 μ g10B is equivalent to each cell and at least contains 109It is a10B)；(2) normal tissue
Relatively low intake (tumor tissues103 times higher than surrounding tissue and blood of B concentration or more).It goes through in BNCT clinical test at present
There are two types of the boron agent only used, i.e. polyhedral boranes (BSH) and 4- boron-L-phenylalanine (BPA), but the cancer target of the two
Effect is undesirable, about boron capture agent cancer target delivering there are still larger rooms for improvement.
It is generated in BNCT mechanism4He and7Li range is very short, thus10The inner cellular localization of B directly affects curative effect.It is existing
There is research to think, positions in nucleus10The curative effect that B is generated is equal volume in cytoplasm1010 times of B.Apart from tumour cell
Core is closer10B receive to fission after the neutron ray irradiation High Linear energy transmission ray of generation hits the probability of nuclear dna and gets over
It is high, it is easier to kill tumour cell.
Traditional boron capture agent cell nucleus targeting ability is poor, and exploitation has the boron capture agent of cell nucleus targeting, it will into one
Step promotes BNCT oncotherapy effect, has important scientific research meaning and application prospect.
Summary of the invention
The characteristics of being greatly facilitated BNCT curative effect for shortcoming existing for this field and the positioning distribution of nucleus boron, this
Invention provides a kind of boracic preparation for having cell nucleus targeting, while showing preferable tumour cell intake, also
It can further target and be positioned at neoplastic cell nuclei, be the Novel boron trapping agent for having cell nucleus targeting.
A kind of boracic preparation having cell nucleus targeting, by 1- bromomethyl neighbour carborane (CB-Br) and contains active ammonia
The cell nucleus targeting drug of base forms；
Alternatively, being made of 1- bromomethyl neighbour carborane, 2 generation dendroid polyamide (PAMAM) and hyaluronic acid (HA)；
Alternatively, by 4- boron-L-phenylalanine (BPA), di-tert-butyl dicarbonate [(BOC)2O] and 2 generation dendroid polyamide
Composition；
Alternatively, being made of adjacent carborane (CB) and dendroid polyethyleneimine (PEI).
1- bromomethyl neighbour's carborane is a kind of polyhedral borine, and boron content is high, and safety is good, is had applied to BNCT's
Good prospect.
The boracic preparation for having cell nucleus targeting in the present invention overcomes traditional weak targeting of boracic preparation organelle
Boron element can be delivered in nucleus by difficulty, greatly promote boron neutron capture therapy effect.
Invention further provides the preparation method of the boracic preparation for having cell nucleus targeting described in one kind, the tools
The boracic preparation of detailed karyon targeting is by 1- bromomethyl neighbour carborane and the cell nucleus targeting medicine group containing active amino
At, comprising steps of
(A-1) the cell nucleus targeting drug containing active amino is dissolved in dimethyl sulfoxide (DMSO), three second is added
Amine stirs 2~4h, and 1- bromomethyl neighbour's carborane is added, continues 48~60h of stirring, adds ether and generates sediment；
(A-2) sediment is collected into bag filter, the use of dimethyl sulfoxide is dialyzate 24~30h of dialysis, is then used
Distilled water is dialyzate 48~60h of dialysis, and decompression is freeze-dried up to the boracic preparation for having cell nucleus targeting.
Preferably, in step (A-1), the cell nucleus targeting drug, triethylamine and 1- bromomethyl neighbour's carborane
Molar ratio be 1:1~3:1~3；
In step (A-2), the molecular cut off of the bag filter is 300~500.
Preferably, the cell nucleus targeting drug containing active amino is Doxorubicin (DOX).Doxorubicin
It is a kind of cytotoxicity series antineoplastic medicament, can be targeted very well after penetrating cell and be positioned at nucleus, base occurs with DNA and inserts
Enter effect and form compound, the processes such as interference DNA synthesis reach tumor-killing effect.Doxorubicin and 1- bromomethyl neighbour's carborane
Boracic Doxorubicin preparation (DOXCB) is formed by interaction, enters preparation targeting nucleus after born of the same parents, it is former to improve boron in nucleus
Sub- content enhances BNCT curative effect in cell nucleus targeting level, while Doxorubicin structure has chemotherapy of tumors effect.How soft boracic is
It is the boron capture agent for having both cell nucleus targeting and Chemotherapy, in percentage by weight, boron atom accounting 4% than star preparation
~8%.
Boracic Doxorubicin preparation can be modified further or is wrapped in pharmaceutical preparation, further increased it and applied in vivo
Targeting, biocompatibility and stability.
Invention further provides the preparation method of the boracic preparation for having cell nucleus targeting described in one kind, the tools
The boracic preparation of detailed karyon targeting is made of 1- bromomethyl neighbour carborane, 2 generation dendroid polyamide and hyaluronic acid, including
Step:
(B-1) 2 generation dendroid polyamide, 1- bromomethyl neighbour carborane and triethylamine are added in dimethyl sulfoxide, stirring
48~60h is added ether and generates sediment；
(B-2) sediment is collected into bag filter, the use of distilled water is dialyzate 72~75h of dialysis, then ultrasound, mistake
Filter obtains dendrimer, dendritic polymer boron capture agent after filtrate decompression freeze-drying；
(B-3) dendrimer, dendritic polymer boron capture agent is soluble in water, be added hyaluronic acid, stand 0.5~1h after ultrasound 0.5~
1h obtains clear aqueous solution, as the boracic preparation for having cell nucleus targeting.
By the modification of 1- bromomethyl neighbour's carborane in dendrimer, dendritic polymer 2 generation dendroid polyamide outer surface, PAMAM-CB is formed
(i.e. the dendrimer, dendritic polymer boron capture agent).The outer surface PAMAM-CB and hyaluronic acid (HA) pass through positive and negative charge sucking action
It is formed nanometer boron capture preparation HA/PAMAM-CB (i.e. final product boracic preparation), enhances vivo applications stability and targeting.
The PAMAM-CB boron capture agent partial size is small, surface lotus positive electricity, easily penetrates into nucleus.
The HA/PAMAM-CB relies on the high-affinity of HA CD44 albumen highly expressed to Partial tumors cell surface,
Further increase the tumor-targeting that PAMAM-CB is applied in vivo；With part PAMAM-CB positive surface charge in HA, HA/ is improved
PAMAM-CB is in sanguimotor stability and compatibility.
Preferably, the molar ratio of the 2 generation dendroid polyamide, 1- bromomethyl neighbour carborane and triethylamine is 1:6
~12:6~12；
In step (B-2), the molecular cut off of the bag filter is 500~2000, and the filtering uses 0.22 μm of filter membrane；
In step (B-3), the mass ratio of the dendrimer, dendritic polymer boron capture agent and hyaluronic acid is 0.5~2.
The molecular weight of the 2 generation dendroid polyamide is 3256, and there are 16 amino in end.
In a preferred example, the PAMAM-CB molecular end modifies 2~4 CB.
Preferably, the average molecular weight of the HA is 8000~10000.
Invention further provides the preparation method of the boracic preparation for having cell nucleus targeting described in one kind, the tools
The boracic preparation of detailed karyon targeting is by 4- boron-L-phenylalanine, di-tert-butyl dicarbonate and 2 generation dendroid polyamide groups
At, comprising steps of
(C-1) di-tert-butyl dicarbonate and 4- boron-L-phenylalanine are dissolved in the mixed solvent of tetrahydrofuran and distilled water
In, stirring 6~for 24 hours, rotary evaporation removes tetrahydrofuran, obtains the aqueous solution containing product, and sodium bicarbonate is added and adjusts pH to 8.5
~9.5, it is extracted with ethyl acetate, rotary evaporation removes ethyl acetate up to product I；
(C-2) product I is dissolved in distilled water, with salt acid for adjusting pH to 4.5~5.5,1- (3- dimethylamino third is added
Base) -3- ethyl carbodiimide and n-hydroxysuccinimide, it stirs and sodium hydroxide is added after 2~4h adjusts pH to 7.5~8.5,
2 generation dendroid polyamide are added, continue 36~48h of stirring, the use of distilled water is then dialyzate 48~72h of dialysis, depressurizes cold
Freeze and is drying to obtain the boracic preparation for having cell nucleus targeting.
4- boron-L-phenylalanine molecule active amino structure is protected using di-tert-butyl dicarbonate, obtains BPABOC (i.e.
Product I).By BPABOC modification in 2 generation dendroid polyamide outer surfaces, the boracic preparation for having cell nucleus targeting is formed
PAMAM-BPABOC。
The PAMAM-BPABOC boron capture agent partial size is small, relies on PAMAM positive surface charge penetrating cell and enters cell
Core.
PAMAM-BPABOC can be modified further or is wrapped in pharmaceutical preparation, further increase its target applied in vivo
Tropism, biocompatibility and stability.
Preferably, in step (C-1), the volume ratio of the in the mixed solvent, tetrahydrofuran and distilled water is 1~2:
1。
Preferably, the di-tert-butyl dicarbonate and 4- boron-L-phenylalanine molar ratio are 1 in step (C-1)
~2:1.
Preferably, in step (C-2), described product I, 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide, N-
The molar ratio of HOSu NHS and 2 generation dendroid polyamide is 6~12:12~24:12~24:1.
Preferably, the dialysis bag retention molecular weight used of dialysing is 1000~3000 in step (C-2).
In a preferred example, the PAMAM-BPABOC molecular end modifies 2~4 BPABOC.
Invention further provides the preparation methods of the boracic preparation for having cell nucleus targeting described in one kind, i.e., ultrasonic
Method, the boracic preparation for having cell nucleus targeting are made of adjacent carborane and dendroid polyethyleneimine, comprising steps of
Dendroid polyethyleneimine is dissolved in ultrapure water, adjacent carborane is added, stirs 12~16h after 8~10h of ultrasound,
Centrifuging and taking supernatant is dialyzate 48~60h of dialysis using distilled water, finally crosses 0.45 μm of water system filter membrane, gained filtrate is
The boracic preparation for having cell nucleus targeting.
Invention further provides the preparation methods of the boracic preparation for having cell nucleus targeting described in one kind, that is, dialyse
Method, the boracic preparation for having cell nucleus targeting are made of adjacent carborane and dendroid polyethyleneimine, comprising steps of
Dendroid polyethyleneimine (PEI) and adjacent carborane (CB) are dissolved in n,N-Dimethylformamide (DMF), stirred
Ultrapure water is added after mixing 8~10h, uses distilled water for dialyzate 48~60h of dialysis after continuing 12~16h of stirring, finally mistake
0.45 μm of water system filter membrane, gained filtrate are the boracic preparation for having cell nucleus targeting.
Invention further provides the preparation methods of the boracic preparation for having cell nucleus targeting described in one kind, i.e. revolving water
Change method, the boracic preparation for having cell nucleus targeting is made of adjacent carborane and dendroid polyethyleneimine, including step
It is rapid:
Dendroid polyethyleneimine and adjacent carborane are dissolved in chloroform, after rotary evaporation eliminates chloroform, are added ultrapure
Water hydratable using distilled water is that dialyzate is dialysed 8~60h after 2~3h of ultrasound, finally crosses 0.45 μm of water system filter membrane, gained filtrate
The as boracic preparation for having cell nucleus targeting.
Ultrasonic method, dialysis and the revolving aquation method is all that adjacent carborane and polyethyleneimine are passed through non-covalent bond
It is formed boron capture agent PEICB (i.e. final product boracic preparation).
In the ultrasonic method, dialysis and revolving aquation method, preferably, point of the dendroid polyethyleneimine
Son amount is 10000~30000；
The molar ratio of the dendroid polyethyleneimine and adjacent carborane is 1:25~100；
The dialysis bag retention molecular weight used of dialysing is 8000~14000.
The present invention also provides the boracic preparations for having cell nucleus targeting described in one kind to control preparing boron neutron absorption
The application in drug is treated, the boracic preparation for having cell nucleus targeting contains10B。
Compared with prior art, the present invention major advantage includes:
(1) the boracic preparation prepared is distributed into born of the same parents in addition to absorbing, can also travel further into nucleus；
(2) the boracic preparation prepared also has chemotherapeutic treatment effect in addition to BNCT curative effect；
(3) the boracic preparation prepared can play bigger BNCT curative effect in same boron content intracellular；
(4) the boracic preparation prepared, biocompatibility is good, and further modification can enhance vivo applications stability and target group
Knit taxis.
Detailed description of the invention
Fig. 1 is DOX, CB-Br and DOXCB's of embodiment 11H-NMR figure；
Fig. 2 is DOX, CB-Br and DOXCB's of embodiment 111B-NMR figure；
Fig. 3 is PAMAM-CB the and HA/PAMAM-CB particle size results figure of embodiment 2；
Fig. 4 is PAMAM-CB the and HA/PAMAM-CB current potential result figure of embodiment 2；
Fig. 5 is CB-Br, PAMAM and PAMAM-CB's of embodiment 21H-NMR figure；
Fig. 6 is BPA, (BOC) of embodiment 32O, PAMAM and PAMAM-BPABOC1H-NMR figure；
Fig. 7 is the particle size results figure of PAMAM (G2) BPABOC, PAMAM (G2) and PAMAM-CB (G4) of embodiment 3；
Fig. 8 be embodiment 3 PAMAM (G2) BPABOC, PAMAM (G2) and PAMAM-CB (G4) pH be 4 and 7.4 two kind
Under the conditions of current potential result figure；
Fig. 9 is the transmission electron microscope picture of the PEICB4 and PEICB5 of embodiment 4；
Figure 10 is the cytotoxicity figure of DOX, CB-Br and DOXCB to GL261 cell of application examples 1；
Figure 11 be application examples 2 CB-Br in GL261 cell boron content figure；
Figure 12 be application examples 2 DOXCB in GL261 cell caryoplasm boron deregulation figure；
Figure 13 be application examples 3 DOXCB in C6 cell caryoplasm boron deregulation figure；
Figure 14 is the DOX and DOXCB of application examples 4 in GL261 cell caryoplasm fluorescence distribution figure；
Figure 15 is the DOX and DOXCB of application examples 4 in C6 cell caryoplasm fluorescence distribution figure；
Figure 16 is cytotoxicity figure of the PAMAM to GL261 cell of application examples 5；
Figure 17 is cytotoxicity figure of the HA to GL261 cell of application examples 5；
Figure 18 is PAMAM (the G2)-FITC of application examples 6 in C6 and U87 cell caryoplasm fluorescence distribution figure；
Figure 19 is Coumarin 6, PAMAMBPABOC/Coumarin 6, the PAMAM (G2)/Coumarin 6 of application examples 7
With PAMAM (G4)-FITC in U87 cell caryoplasm fluorescence distribution figure；
Figure 20 is the PEICB1-FITC of application examples 8 in U87 cell caryoplasm fluorescence distribution figure；
Figure 21 is the PEICB5-FITC of application examples 8 in U87 cell caryoplasm fluorescence distribution figure；
Figure 22 is the PEI-FITC of application examples 8 in U87 cell caryoplasm fluorescence distribution figure.
Specific embodiment
With reference to the accompanying drawing and specific embodiment, the present invention is further explained.It should be understood that these embodiments are merely to illustrate
The present invention rather than limit the scope of the invention.The operating method of actual conditions is not specified in the following example, usually according to
Normal condition, or according to the normal condition proposed by manufacturer.
1 boracic Doxorubicin preparation (DOXCB) of embodiment
(A-1) Doxorubicin (i.e. doxorubicin hydrochloride) is dissolved in anhydrous dimethyl sulphoxide, is added under the conditions of nitrogen protection
Triethylamine stirs 2h, and 1- bromomethyl neighbour's carborane is added, and magneton stirs 48h at room temperature, adds excess diethyl ether, makes to have precipitated
Entirely；The molar ratio of Doxorubicin, triethylamine and 1- bromomethyl neighbour's carborane is 1:2:1；
(A-2) it collects in the bag filter that sediment is 500 to molecular cut off, the use of dimethyl sulfoxide is dialyzate dialysis
For 24 hours, then using distilled water is dialyzate dialysis 48h, and decompression freeze-drying obtains red powder, as boracic Doxorubicin system
Agent (DOXCB).
The DOXCB of the present embodiment is obtained after being cleared up with concentrated nitric acid by inductivity coupled plasma mass spectrometry (ICP-MS) detection
Boron content is 5.15wt%.
DOX and DOXCB makees solvent with deuterated DMSO, and CB-Br makees solvent dissolution with deuterated chloroform and carries out1H-NMR analysis.
As shown in Figure 1,1H-NMR the result shows that: the characteristic peak of DOX is substantially identical as DOXCB signal peak, and the feature B-H of CB-Br
Signal peak (1.5-3.0ppm) also occurs in DOXCB signal peak, it was demonstrated that the successful synthesis of DOXCB.
DOX and DOXCB makees solvent with deuterated DMSO, and CB-Br makees solvent dissolution with deuterated chloroform and carries out11B-NMR analysis.
As shown in Fig. 2,11B-NMR the result shows that: the basic no signal peak DOX, DOXCB have apparent boron signal peak, with CB-Br feature boron
Signal peak shift is almost the same, it was demonstrated that the successful synthesis of DOXCB.
2 nanometers of boron capture preparations (HA/PAMAM-CB) of embodiment
(B-1) by 2 generation dendroid polyamide, 1- bromomethyl neighbour carborane and triethylamine, diformazan is added in 1:1:12 in molar ratio
In base sulfoxide, after room temperature magneton stirs 48h, it is complete to precipitating that excess diethyl ether is added；
(B-2) it collects in the bag filter that sediment is 1000 to molecular cut off, the use of distilled water is dialyzate dialysis
Then 72h crosses 0.22 μm of filter membrane, obtains dendrimer, dendritic polymer after filtrate decompression is freeze-dried after 200W power probe ultrasound 3min
Boron capture agent PAMAM-CB；
(B-3) dendrimer, dendritic polymer boron capture agent is soluble in water, the hyaluronic acid of the quality such as addition stands water-bath after 0.5h
Ultrasonic 0.5h, then 200W power probe ultrasound 2min obtains clear aqueous solution, obtains clarification HA/PAMAM-CB aqueous solution.
The PAMAM-CB of the present embodiment detects to obtain boron content to be 9.33wt% after being cleared up with concentrated nitric acid by ICP-MS.
PAMAM-CB the and HA/PAMAM-CB ultra-pure water solution of the present embodiment uses Malvern Zetasizer Nano
ZS90 series laser particle size analyzer measures particle diameter distribution situation, as shown in figure 3, PAMAM-CB about 664.7nm, PDI is about
0.856, dispersibility is bad；HA/PAMAM-CB partial size about 293.0nm, PDI about 0.141, dispersibility is preferably.HA wraps up lower boron prisoner
Obtain that agent size is more regular, and partial size is smaller.
PAMAM-CB the and HA/PAMAM-CB ultra-pure water solution of the present embodiment uses Malvern Zetasizer Nano
ZS90 series of electrical potentials analysis-e/or determining Potential distribution situation, as shown in figure 4, the surface PAMAM-CB about 3.427mV, lotus positive potential；
The surface HA/PAMAM-CB about -0.029mV, bear current potential.HA wraps up lower boron capture agent current potential and is changed into negative potential, enhances body
Interior blood circulation stability.
CB-Br deuterated chloroform, PAMAM and PAMAM-CB use deuterated water as solvent, carry out1H-NMR analysis.Such as Fig. 5
It is shown,1H-NMR the result shows that: there is the characteristic signal peak of CB-Br and PAMAM simultaneously in PAMAM-CB signal peak, it was demonstrated that
The successful synthesis of PAMAM-CB.
3 dendrimer, dendritic polymer boron capture agent (PAMAM-BPABOC) of embodiment
(C-1) by di-tert-butyl dicarbonate and 4- boron-L-phenylalanine, 1:1 is dissolved in tetrahydrofuran and distilled water in molar ratio
Volume ratio is the in the mixed solvent of 1:1, and for 24 hours, rotary evaporation removes tetrahydrofuran for stirring, obtains the aqueous solution containing product, is added
0.1M sodium bicarbonate aqueous solution adjusts pH to 9, is extracted with ethyl acetate, and rotary evaporation removes ethyl acetate up to product I；
(C-2) product I is dissolved in distilled water, adjusts pH to 5 with 1M aqueous hydrochloric acid solution, 1- (3- dimethylamino third is added
Base) -3- ethyl carbodiimide and n-hydroxysuccinimide, stir and 1M sodium hydrate aqueous solution is added after 4h adjusts pH to 8, add
Enter 2 generation dendroid polyamide, continues to stir 48h；Described product I, 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide,
N-hydroxysuccinimide and 2 generation dendroid polyamide feed intake in molar ratio for 6:10:10:1；
(C-3) acquired solution is fitted into the bag filter that molecular cut off is 1000, the use of distilled water is dialyzate dialysis
48h, decompression are freeze-dried up to dendrimer, dendritic polymer boron capture agent (PAMAM-BPABOC).
The PAMAM-BPABOC of the present embodiment cleared up with concentrated nitric acid after by ICP-MS detect boron content, boron content is about
1.0wt%, i.e. 1 PAMAM molecule graft about 3 BPABOC molecules.
BPA、(BOC)2O, it is carried out after PAMAM and PAMAM-BPABOC is dissolved with deuterated solvent1H-NMR analysis.Such as Fig. 6 institute
Show,1H-NMR the result shows that DOXCB successful synthesis.
PAMAM (G2)-is measured using Malvern Zetasizer Nano ZS90 series laser particle size analyzer
The particle diameter distribution of BPABOC, PAMAM (G2) (2 generation dendroid polyamide) and PAMAM (G4) (4 generation dendroid polyamide) aqueous solution
Situation.As shown in fig. 7, PAMAM (G2) partial size is minimum, about 43.9nm；Since BPABOC, PAMAM (G2)-are grafted in outer surface
BPABOC partial size ratio PAMAM (G2) is slightly larger, about 57.0nm；And PAMAM (G4) molecular weight greatly and easily gather due to partial size most
Greatly, about 140.4nm.
Respectively under conditions of pH is 4.0 and 7.4, using Malvern Zetasizer Nano ZS90 series laser electricity
The surface potential situation of position analysis-e/or determining PAMAM (G2)-BPABOC, PAMAM (G2) and PAMAM (G4) aqueous solution.Such as Fig. 8 institute
Show, under the conditions of pH is 7.4, PAMAM (G2)-BPABOC, PAMAM (G2) and PAMAM (G4) surface potential are essentially electroneutral；
And under the conditions of pH is 4.0, PAMAM (G2)-BPABOC, PAMAM (G2) and PAMAM (G4) surface potential fundamental transformation be it is electric just
Property, this prompt preparation can play proton pump effect escape lysosome in the lysosome of strong acid in the cell.
4 dendroid polyethyleneimine boron capture agent (PEICB) of embodiment
Has nucleus rich in electropositive material PEI and carborane preparation using non-covalent bond (hydrogen bond, two hydrogen bonds) connection
Three kinds of methods are mainly applied in targeting boron agent.
(1) ultrasonic method: 69.32mg PEI (molecular weight 25000) water bath sonicator is taken to be dissolved in 20mL ultrapure water.It adds
20mg neighbour carborane CB, magneton stirs 12 hours after water bath sonicator 8 hours, centrifuging and taking supernatant, is packed into molecular cut off and is
In 8000~14000 bag filter, the use of distilled water is that dialyzate is dialysed 2 days, finally crosses 0.45 μm of moisture film, obtain solution, remember
For PEICB1.
It is separately 40mg by same procedure but increase CB amount, obtains solution and be denoted as PEICB2.
The clarification of PEICB1 and PEICB2 solution, there is slight Tyndall effect.
(2) dialysis: 1 equivalent PEI (69.32mg) and 50 equivalent CB (20mg) is taken to be dissolved in 15mL DMF, room temperature magnetic
Son stirring 8 hours after be added dropwise in 10mL ultrapure water, stirring 12 hours after be packed into molecular cut off be 8000~14000 it is saturating
It analyses in bag, the use of distilled water is that dialyzate is dialysed 2 days, finally crosses 0.45 μm of moisture film, obtain solution, be denoted as PEICB3.
Separately by same procedure but increases CB amount and be denoted as PEICB4 to obtain solution after 40mg.
The clarification of PEICB3 and PEICB4 solution, there is slightly obvious Tyndall effect.
(3) it rotates aquation method: 1 equivalent PEI (34mg) and 50 equivalent CB (10mg) being taken to be dissolved in 50mL chloroform, room temperature rotation
2 hours removing chloroforms are steamed, the ultrapure water hydratable of 25mL, after water bath sonicator 2 hours, Probe Ultrasonic Searching 5 minutes (power 300W, works is added
Make 3 seconds, rest 2 seconds), it is fitted into the bag filter that molecular cut off is 8000~14000 after obtaining turbid solution, uses distilled water
For dialyzate dialysis 8 hours, 0.45 μm of moisture film is finally crossed, solution is obtained, is denoted as PEICB5, still some are muddy, Tyndall effect
Obviously.Not crossing 0.45 μm of moisture film solution takes 2mL to keep on file, is denoted as PEICB6.Weighing PEI, to be made into 10mg/mL PEI deionization water-soluble
Liquid is denoted as PEI.
The preparation of the present embodiment by ICP-MS detects boron content after being cleared up with concentrated nitric acid, and 1 PEI molecule is calculated and connects
Meet about 6 CB.
The preparation of the present embodiment measures grain with Malvern Zetasizer Nano ZS90 series laser particle size analyzer
Diameter, the results show that PEICB1 partial size about 92.19nm, PEICB2 partial size about 111.53nm, PEICB3 partial size about 120.8nm,
PEICB4 partial size about 80.14nm, PEICB5 partial size about 123.60nm, PEICB6 partial size about 185.50nm, PEI partial size is about
6.99nm illustrates that PEI preparation modification CB partial size becomes larger, and about 100nm is suitble to the administration of tumour nano target.
The preparation of the present embodiment measures surface with Malvern Zetasizer Nano ZS90 series laser potentiometric analyzer
Current potential, as the result is shown: PEICB1 current potential about 0.43mV, PEICB2 current potential about 0.79mV, PEICB3 current potential about 4.72mV, PEICB4
Current potential about 5.66mV, PEICB5 current potential about 27.03mV, PEICB6 current potential about 43.67mV, PEI current potential about 3.82mV illustrate PEI
Current potential is still positive after preparation modification CB, and there are still enter cell nuclear capability.
The PEICB4 and PEICB5 of the present embodiment shoot pattern with transmission electron microscope, as shown in figure 9, figure A is PEICB4 sample
Figure, figure B are PEICB5 sample drawing, and two sample regular appearances are slightly rounded, and partial size about 100nm and particle instrument measurement result are basic
Unanimously.
Cytotoxicity experiment of the 1 boracic Doxorubicin preparation (DOXCB) of application examples to GL261 cell
For the GL261 cell inoculation of logarithmic growth phase in 96 orifice plates, every hole cell quantity is 3 × 103, at 37 DEG C, 5%
CO2It is cultivated in cell incubator and withdraws from culture solution afterwards for 24 hours, 200 μ L drug containing culture solution (DOX, CB-Br and DOX- are added in every hole
CB).DOX and DOX-CB is respectively provided with 8 equal specific concentrations gradients, and CB-Br group is arranged 6 and waits specific concentrations gradient, and each concentration 6 multiple
Hole, DOX group maximum concentration are that 20 μ g/mL, CB-Br group maximum concentrations are 200 μM, and DOX-CB group maximum concentration is 80 μ g/mL, are set
Continue to withdraw from culture solution after being incubated for 48h in cell incubator, every hole is added 100 culture solutions of the μ L containing 10%CCK8, continues to train
2min is vibrated on the oscillator after supporting 4h, measures OD value under 450nm wavelength using microplate reader, and cells survival is calculated using formula
Rate evaluates cytotoxicity:
As shown in Figure 10, CB-Br is substantially to GL261 cytotoxic, and DOX and DOXCB is incubated for GL261 cell 48h's
IC50 value is respectively 0.00080178 μ g/mL and 19.23 μ g/mL, prompts GL261 cell very sensitive to DOX, due to DOX and CB-
It is combined between Br and affects drug toxicity, so that DOXCB much smaller to GL261 toxicity ratio DOX, but there are still certain tumour cells
Fragmentation effect.DOXCB is the boron capture agent for having chemotherapy of tumors effect.
2 boracic Doxorubicin preparation (DOXCB) of application examples is in GL261 cell caryoplasm distribution situation
It is every with the assessment of ICP-MS method respectively after 16 μ g/mL are incubated for GL261 cell 6 hours that DOXCB, which is arranged, to be incubated for concentration
Million intracellular boron intakes (n=3), while being commented after application cell core extracts kit acquisition nucleus with ICP-MS method
Estimate boron intake (n=3) in every million nucleus.Simultaneously with CB-Br (0.8 μ g/mL) incubated cell of identical boron mole
As a control group.
As shown in figure 11, CB-Br base following figure result can be seen that GL261 cell absorbs control formulation CB-Br essentially boron-free,
And have DOXCB preparation greater than 20ng B/106Cells intake (Figure 12), wherein the boron element of more than half is located at nucleus
Interior, preparation effect meets desired design, is suitble to further be wrapped in nanometer formulation and realizes in-vivo tumour targeted delivery.
3 boracic Doxorubicin preparation (DOXCB) of application examples is in C6 cell caryoplasm distribution situation
It is every with the assessment of ICP-MS method respectively after 16 μ g/mL are incubated for GL261 cell 6 hours that DOXCB, which is arranged, to be incubated for concentration
Million intracellular boron intakes (n=3), while being commented after application cell core extracts kit acquisition nucleus with ICP-MS method
Estimate boron intake (n=3) in every million nucleus.
As shown in figure 13, C6 cell has greater than 20ng B/10 DOXCB preparation6Cells intake, wherein more than half
Boron be located in nucleus, preparation effect meets desired design, is suitble to further be wrapped in nanometer formulation and realizes in-vivo tumour
Targeted delivery.
4 boracic Doxorubicin preparation (DOXCB) of application examples is in GL261 and C6 cell caryoplasm fluorescence distribution situation
It is after 16 μ g/mL are incubated for GL261 and C6 cell 6h respectively, to be rinsed cell 3 times with PBS that setting DOXCB, which is incubated for concentration,
4% paraformaldehyde room temperature fixes 0.5h, and DAPI dye marker nucleus, shooting record DOXCB exists under Laser Scanning Confocal Microscope
GL261 and C6 cell caryoplasm fluorescence distribution figure.DOX control group is set simultaneously.
As shown in figure 14, DOX red fluorescence is substantially distributed in GL261 nucleus, and DOXCB is then largely positioned at carefully
In karyon, small part is in cytoplasm.Figure 15 illustrates that DOX red fluorescence is substantially distributed in C6 nucleus, and the then big portion DOXCB
Divide and be positioned in nucleus, small part is in cytoplasm.DOXCB shows consistent result in both the above tumor cell line:
It can enter in nucleus, and then deliver boron and enter nucleus, enhance BNCT effect.
Cytotoxicity experiment of the 5 nanometers of boron capture preparations (HA/PAMAM-CB) of application examples to GL261 cell
For the GL261 cell inoculation of logarithmic growth phase in 96 orifice plates, every hole cell quantity is 3 × 103, at 37 DEG C, 5%
CO2It is cultivated in cell incubator and withdraws from culture solution afterwards for 24 hours, 200 μ L drug containing culture solutions (PAMAM and HA) are added in every hole, are respectively provided with 8
A equal specific concentrations gradient, 6 multiple holes of each concentration, maximum concentration are 800 μM, are placed in cell incubator after continuing to be incubated for 48h
Culture solution is removed, 100 culture solutions of the μ L containing 10%CCK8 are added in every hole, continue to vibrate 2min on the oscillator after cultivating 4h, make
OD value is measured under 450nm wavelength with microplate reader, and cells survival rate is calculated using formula and evaluates cytotoxicity:
As shown in figure 16, PAMAM is substantially non-toxic to GL261 cell under low concentration, and due to PAMAM positive electricity under high concentration
Lotus causes bigger toxicity to cell, and IC50 value is 351.48 μM.As shown in figure 17, HA is substantially non-toxic to GL261 cell, with
This reduces its periphery toxicity using HA package PAMAM-CB, and it is thin to CD44 high expression tumour also to can increase boron capture agent targeted delivery
Born of the same parents (such as U87, C6, GL261, A549).
6 nanometers of boron capture preparations (HA/PAMAM-CB) of application examples are in C6 and U87 cell caryoplasm fluorescence distribution situation
PAMAM is dissolved in PBS (pH=7.4), investment FITC dialyses 12h after reaction overnight, obtains FITC label
PAMAM (G2) is denoted as PAMAM (G2)-FITC, chooses U87 and C6 cell respectively and does model cell.FITC control group concentration is 1 μ
G/mL, PAMAM (G2)-FITC concentration is 20 μ g/mL, incubation time have 6h, 12h and for 24 hours, cell it is fixed after DAPI dye marker
Nucleus assesses PAMAM in cell caryoplasm distribution situation.
As shown in figure 18, PAMAM (G2)-FITC enters born of the same parents in 6h to enter core obvious, but 12h and under fluorescent brightness is obvious for 24 hours
Drop, may be susceptible to by cell outlet.Show that partial size PAMAM less than normal (G2) is suitable as cell nucleus targeting preparations carrier, still
It should also be noted that preparation enters nucleus, there are window times in application process.
7 dendrimer, dendritic polymer boron capture agent (PAMAM-BPABOC) of application examples is in U87 cell caryoplasm fluorescence distribution situation
PAMAM-BPABOC, PAMAM (G2) and PAMAM (G4) physics contain fluorescent molecule coumarin 6, respectively obtain
PAMAMBPABOC/Coumarin 6, PAMAM (G2)/Coumarin6 and PAMAM (G4)/Coumarin 6.It is incubated for U87 respectively
It is rinsed 3 times after cell 2h with PBS, DAPI dye marker nucleus after the fixed cell of 4% paraformaldehyde, is protected under Laser Scanning Confocal Microscope
Picture is deposited, assesses each preparation in cell caryoplasm distribution situation.
As shown in figure 19, partial size preparation PAMAMBPABOC/Coumarin 6 less than normal and PAMAM (G2)/Coumarin 6
Fluorescence is stronger in nucleus, be prompted into nuclear capability ratio Coumarin 6 dissociate fluorescer control and PAMAM (G2)/Coumarin 6
Preparation is strong.
8 dendroid polyethyleneimine boron capture agent (PEICB) of application examples is in U87 cell caryoplasm fluorescence distribution situation
PEICB1, PEICB5 and PEI in embodiment 4 are dissolved separately in PBS (pH=7.4), and investment FITC reaction is overnight
After dialyse 12h, obtain the preparation of FITC label, be denoted as PEICB1-FITC, PEICB5-FITC and PEI-FITC, it is thin to choose U87
Born of the same parents do model cell.FITC control group concentration is that 1 μ g/ml, PEICB1-FITC and PEICB5-FITC is 20 μ g/ml, is incubated for
Time have 6h, 12h and for 24 hours, cell it is fixed after DAPI dye marker nucleus, assess preparation in cell caryoplasm fluorescence distribution feelings
Condition.
As shown in Figure 20~22, PEICB1-FITC (Figure 20), PEICB5-FITC (Figure 21) and PEI-FITC (Figure 22),
Have in fluorescence distribution nucleus, this illustrates that PEI can be used as cell nucleus targeting boron capture agent carrier.
In addition, it should also be understood that, those skilled in the art can be to this hair after having read foregoing description content of the invention
Bright to make various changes or modifications, these equivalent forms also fall within the scope of the appended claims of the present application.
Claims (10)
1. a kind of boracic preparation for having cell nucleus targeting, which is characterized in that by 1- bromomethyl neighbour carborane and containing active
The cell nucleus targeting drug of amino forms；
Alternatively, being made of 1- bromomethyl neighbour carborane, 2 generation dendroid polyamide and hyaluronic acid；
Alternatively, being made of 4- boron-L-phenylalanine, di-tert-butyl dicarbonate and 2 generation dendroid polyamide；
Alternatively, being made of adjacent carborane and dendroid polyethyleneimine.
2. a kind of preparation method of the boracic preparation according to claim 1 for having cell nucleus targeting, which is characterized in that
The boracic preparation for having cell nucleus targeting is by 1- bromomethyl neighbour carborane and containing the cell nucleus targeting of active amino
Property drug composition, comprising steps of
(A-1) the cell nucleus targeting drug containing active amino is dissolved in dimethyl sulfoxide, addition triethylamine, stirring 2~
4h is added 1- bromomethyl neighbour's carborane, continues 48~60h of stirring, adds ether and generates sediment；
(A-2) sediment is collected into bag filter, the use of dimethyl sulfoxide is dialyzate 24~30h of dialysis, then using distillation
Water is dialyzate 48~60h of dialysis, and decompression is freeze-dried up to the boracic preparation for having cell nucleus targeting.
3. the preparation method of the boracic preparation according to claim 2 for having cell nucleus targeting, which is characterized in that step
(A-1) in, the molar ratio of the cell nucleus targeting drug, triethylamine and 1- bromomethyl neighbour's carborane is 1:1~3:1~3；
In step (A-2), the molecular cut off of the bag filter is 300~500.
4. a kind of preparation method of the boracic preparation according to claim 1 for having cell nucleus targeting, which is characterized in that
The boracic preparation for having cell nucleus targeting is by 1- bromomethyl neighbour carborane, 2 generation dendroid polyamide and hyaluronic acid
Composition, comprising steps of
(B-1) 2 generation dendroid polyamide, 1- bromomethyl neighbour carborane and triethylamine are added in dimethyl sulfoxide, stirring 48~
60h is added ether and generates sediment；
(B-2) sediment is collected into bag filter, the use of distilled water is dialyzate 72~75h of dialysis, then ultrasound, filtering, filter
Dendrimer, dendritic polymer boron capture agent is obtained after liquid decompression freeze-drying；
(B-3) dendrimer, dendritic polymer boron capture agent is soluble in water, hyaluronic acid is added, stands 0.5~1h of ultrasound after 0.5~1h,
Obtain clear aqueous solution, as the boracic preparation for having cell nucleus targeting.
5. the preparation method of the boracic preparation according to claim 4 for having cell nucleus targeting, which is characterized in that described
2 generation dendroid polyamide, 1- bromomethyl neighbour carborane and triethylamine molar ratio be 1:6~12:6~12；
In step (B-2), the molecular cut off of the bag filter is 500~2000, and the filtering uses 0.22 μm of filter membrane；
In step (B-3), the mass ratio of the dendrimer, dendritic polymer boron capture agent and hyaluronic acid is 0.5~2.
6. a kind of preparation method of the boracic preparation according to claim 1 for having cell nucleus targeting, which is characterized in that
The boracic preparation for having cell nucleus targeting is gathered by 4- boron-L-phenylalanine, di-tert-butyl dicarbonate and 2 generation dendroids
Amide composition, comprising steps of
(C-1) di-tert-butyl dicarbonate and 4- boron-L-phenylalanine are dissolved in the in the mixed solvent of tetrahydrofuran and distilled water, stirred
Mix 6~for 24 hours, rotary evaporation removes tetrahydrofuran, obtains the aqueous solution containing product, be added sodium bicarbonate adjust pH to 8.5~
9.5, it is extracted with ethyl acetate, rotary evaporation removes ethyl acetate up to product I；
(C-2) product I is dissolved in distilled water, with salt acid for adjusting pH to 4.5~5.5,1- (3- dimethylamino-propyl) -3- is added
Ethyl carbodiimide and n-hydroxysuccinimide are added sodium hydroxide adjusting pH to 7.5~8.5 after stirring 2~4h, are added 2
For dendroid polyamide, continue 36~48h of stirring, the use of distilled water is then dialyzate 48~72h of dialysis, decompression freeze-drying
Up to the boracic preparation for having cell nucleus targeting.
7. a kind of preparation method of the boracic preparation according to claim 1 for having cell nucleus targeting, which is characterized in that
The boracic preparation for having cell nucleus targeting is made of adjacent carborane and dendroid polyethyleneimine, comprising steps of
Dendroid polyethyleneimine is dissolved in ultrapure water, adjacent carborane is added, 12~16h, centrifugation are stirred after 8~10h of ultrasound
Supernatant is taken, the use of distilled water is dialyzate 48~60h of dialysis, finally crosses 0.45 μm of water system filter membrane, gained filtrate is described
The boracic preparation for having cell nucleus targeting.
8. a kind of preparation method of the boracic preparation according to claim 1 for having cell nucleus targeting, which is characterized in that
The boracic preparation for having cell nucleus targeting is made of adjacent carborane and dendroid polyethyleneimine, comprising steps of
Dendroid polyethyleneimine and adjacent carborane are dissolved in n,N-Dimethylformamide, are added after 8~10h of stirring ultrapure
Water uses distilled water for dialyzate 48~60h of dialysis, finally crosses 0.45 μm of water system filter membrane, gained filter after continuing 12~16h of stirring
Liquid is the boracic preparation for having cell nucleus targeting.
9. a kind of preparation method of the boracic preparation according to claim 1 for having cell nucleus targeting, which is characterized in that
The boracic preparation for having cell nucleus targeting is made of adjacent carborane and dendroid polyethyleneimine, comprising steps of
Dendroid polyethyleneimine and adjacent carborane are dissolved in chloroform, after rotary evaporation eliminates chloroform, ultrapure water water is added
Change, it is dialyzate 8~60h of dialysis that distilled water is used after 2~3h of ultrasound, finally crosses 0.45 μm of water system filter membrane, gained filtrate is
The boracic preparation for having cell nucleus targeting.
10. a kind of boracic preparation according to claim 1 for having cell nucleus targeting is preparing boron neutron capture therapy
Application in drug, which is characterized in that the boracic preparation for having cell nucleus targeting contains10B。
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| US5630786A (en) * | 1994-06-27 | 1997-05-20 | Ionix Corporation | Boron neutron capture enhancement of fast neutron therapy |
| US6696038B1 (en) * | 2000-09-14 | 2004-02-24 | Expression Genetics, Inc. | Cationic lipopolymer as biocompatible gene delivery agent |
| GB0112844D0 (en) * | 2001-05-25 | 2001-07-18 | Psimei Pharmaceuticals Plc | Neutron capture therapy |
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| CN100417421C (en) * | 2006-09-26 | 2008-09-10 | 东南大学 | Organic metal carborane target formulation and its preparation method |
| CN102429870A (en) * | 2011-12-21 | 2012-05-02 | 中国药科大学 | Novel tumor-targeting arboraceous polymer nano carrier of camptothecin drug |
| CN104399094B (en) * | 2014-11-03 | 2017-02-15 | 苏州大学附属第一医院 | Targeted boron preparation and preparation method |
| CN104623685B (en) * | 2015-02-11 | 2018-03-09 | 中国药科大学 | A kind of mesenchyma stroma of tumors pH responsive types targeting dendrimers and preparation method thereof |
| US9795624B2 (en) * | 2015-05-04 | 2017-10-24 | Research Foundation Of The City University Of New York | Cationic polymers as co-drugs for chemotherapeutic agents |
| CN107902642A (en) * | 2017-10-12 | 2018-04-13 | 兰州大学 | A kind of preparation and application study of the graphene quantum dot being imaged for nuclei images and mitochondria |
-
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Non-Patent Citations (2)
| Title |
|---|
| DEEPTHI A ET AL.: "Targeted Drug Delivery to the Nucleus and its Potential Role in Cancer Chemotherapy", 《J. PHARM. SCI. & RES.》 * |
| HEJIAN XIONG ET AL.: "Doxorubicin-Loaded Carborane-Conjugated Polymeric Nanoparticles as Delivery System for Combination Cancer Therapy", 《BIOMACROMOLECULES》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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