CN110179992A - A kind of pair of neogenesis cryptococcus has drug-polymer conjugate of high-affinity and its preparation method and application - Google Patents
A kind of pair of neogenesis cryptococcus has drug-polymer conjugate of high-affinity and its preparation method and application Download PDFInfo
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Abstract
The present invention relates to the drug-polymer conjugates and its preparation method and application that a kind of pair of neogenesis cryptococcus has high-affinity, belong to the polymer arts of pharmaceutical preparation.Drug-polymer conjugate in the present invention is to modify amphotericin B in poly(lactic-co-glycolic acid)-polyethylene glycol-N, and HOSu NHS surface, structure is as shown in general formula I.Drug-polymer conjugate of the invention has high compatibility to neogenesis cryptococcus, has good biocompatibility and biodegradability.After the drug-polymer conjugate is mixed with poly(lactic-co-glycolic acid)-polyethylene glycol, package amphotericin B forms nanoparticle, has good therapeutic effect for infection by Cryptococcus neoformans in fungal infection, especially fungi.
Description
Technical field
The invention belongs to the polymer arts of pharmaceutical preparation, and in particular to a kind of pair of neogenesis cryptococcus has the medicine of high-affinity
Object-polymer conjugates and its preparation method and application.
Background technique
Polymer has many advantages such as can modify, drugloading rate is high, delays drug release and has good stability.Polylactic acid-
Co-glycolic acid-polyethylene glycol [(Poly (lactic-co-glycolic acid)-polyethylene glycol,
PLGA-PEG] be used as representative polymer, have good biological degradability, biocompatibility, it is nontoxic, without teratogenesis, drop
The characteristics such as solution rate and rate of releasing drug are controllable, are always the research hotspot in drug delivery field.
Amphotericin B (Amphotericin B, AMB) is a kind of heptaene, and AMB molecular formula is C47H73NO17, point
Son amount is 924.09, and yellow or orange-yellow powder are almost odorless, have draw it is moist, it is light sensitive.It is hydrophobic that AMB contains a polyenoid
The hydrophilic side chain of side chain and a polyhydroxy, polyenoid side chain can be with the main constituents ergosterol phases on fungal cell membrane
Interaction forms sterol-polyenoid compound, forms many hydrophilic micropores on cell membrane, increase permeability of cell membranes
Add, intracellular small-molecule substance and electrolyte leakage, causes fungal cell dead, be antifungal drug " goldstandard ".It is clinical
On be mainly used for the invalid deep fungal infection of other medicines treatment, anti-leishmaniasis.The structural formula of AMB is as follows:
Therefore the amphotericin B (AMB) that will be specifically bound with the ergosterol on neogenesis cryptococcus cell membrane is as suitable
The ligand modified water-wet side to PLGA-PEG prepares functional nano grain in this, as drug-polymer conjugate, with single material
Material prepares nanoparticle and compares, and while the drug being loaded can be made to recycle in blood, can also increase and be loaded drug in spy
Determine the aggregate amount at position, improves lesion drug concentration, reach better therapeutic effect.
Summary of the invention
In view of this, one of the objects of the present invention is to provide a kind of pair of neogenesis cryptococcus to have the drug-of high-affinity poly-
Close object conjugate；The second object of the present invention is that providing a kind of pair of neogenesis cryptococcus has the drug-polymer of high-affinity even
Join the preparation method of object；The third object of the present invention is to provide a kind of drug-polymer conjugate for being loaded with amphotericin B and receives
The grain of rice；The fourth object of the present invention is to provide a kind of preparation of drug-polymer conjugate nanoparticle for being loaded with amphotericin B
Method；The fifth object of the present invention is to provide a kind of drug-polymer conjugate nanoparticle for being loaded with amphotericin B and is preparing
Application in the pharmaceutical preparation of antimycotic and/or anti-leishmaniasis.
In order to achieve the above objectives, the invention provides the following technical scheme:
1, a kind of pair of neogenesis cryptococcus has the drug-polymer conjugate of high-affinity, the drug-polymer conjugate
General structure it is as follows:
Wherein, the integer that m is 50~80, the integer that n is 65~95.
Preferably, the drug-polymer conjugate includes poly(lactic-co-glycolic acid)-polyethylene glycol-N, hydroxyl
Succinimide and amphotericin B, amphotericin B are modified in the poly(lactic-co-glycolic acid)-polyethylene glycol-N, hydroxyl
Base succinimide surface.
2, said medicine-polymer conjugates preparation method, it is described the preparation method is as follows: by amphotericin B and poly- cream
Acid-co-glycolic acid-polyethylene glycol-N, HOSu NHS (PLGA-PEG-NHS) are added to containing N, and N- bis- is different
In the anhydrous organic solvent of propylethylamine, coupling reaction is carried out, precipitating reagent is added after reaction, solid reaction is obtained by filtration
Object, 20~30 DEG C at a temperature of carry out vacuum drying the drug-polymer for having high-affinity to neogenesis cryptococcus can be obtained
Conjugate.
Preferably, the molar ratio of the amphotericin B and PLGA-PEG-NHS are 1:1~2:1；The polylactic acid-glycolic base
Acetate multipolymer-polyethylene glycol-N, the degree of polymerization of poly lactide-glycolide acid is 50~80 in HOSu NHS,
The degree of polymerization of polyethylene glycol is 65~95.
Preferably, the concentration containing n,N-diisopropylethylamine is 5.0~10.0mmol/L in the anhydrous organic solvent.
Preferably, the organic solvent is any one in DMF, DMSO, methylene chloride or chloroform.
Preferably, the temperature of the coupling reaction is 20~30 DEG C, and the reaction time is 12~35h.
Preferably, the precipitating reagent is any one in ether, isopropyl ether or methanol.
3, a kind of drug-polymer conjugate nanoparticle for being loaded with amphotericin B, the nanoparticle include said medicine-
Polymer conjugates and amphotericin B, the amphotericin B are wrapped up by the drug-polymer conjugate.
4, the preparation method of above-mentioned a kind of polymer conjugates nanoparticle for being loaded with amphotericin B, the preparation method packet
Include following steps:
(1) by poly(lactic-co-glycolic acid)-polyethylene glycol (PLGA-PEG) and described there is high parent to neogenesis cryptococcus
Be mixed to form mixture according to 1~5:1 with the drug-polymer conjugate (PLGA-PEG-AMB) of property, by the mixture with
Amphotericin B is mixed according to the mass ratio of 5~15:1, and DMSO is added and dissolves to form mixed solution；
(2) mixed solution is slowly injected into polyvinyl alcohol (PVA) aqueous solution, and be ultrasonically treated to obtain oil-in-water cream
Liquid；
(3) oil-in-water emulsion is centrifuged, upper layer aaerosol solution is taken to be centrifuged by Superfreezing, collect nano particle,
And with milli-Q water, sediment is obtained；
(4) obtained sediment is suspended in 3~6% aqueous sucrose solutions, excessively 0.22 μm of filter membrane, -60 DEG C~-100 DEG C
Under be freeze-dried to obtain the drug-polymer conjugate nanoparticle for being loaded with amphotericin B.
Preferably, the concentration in polyvinyl alcohol (PVA) aqueous solution described in step (2) is 0.5~1.5%, the concentration
Unit be w/v.
5, application of the above-mentioned nanoparticle in the pharmaceutical preparation for preparing antimycotic and/or anti-leishmaniasis.
Preferably, the pharmaceutical preparation includes external medicine preparation, Sucked medicine preparation or intravenous (IV) drug preparation
In any one.
The beneficial effects of the present invention are:
1, the invention discloses drug-polymer conjugate and its preparation sides that a kind of pair of neogenesis cryptococcus has high-affinity
Method modifies amphotericin B in poly(lactic-co-glycolic acid)-polyethylene glycol-N, and the surface of HOSu NHS is right
Neogenesis cryptococcus has high affinity, good biocompatibility and biodegradability, has very high researching value and answers
With value, have a good application prospect in terms of the pharmaceutical preparation for preparing antimycotic and/or anti-leishmaniasis.
2, the invention also discloses a kind of drug-polymer conjugate nanoparticle for being loaded with amphotericin B and its preparation sides
Method, use it is prepared by the present invention have the drug-polymer conjugate of high-affinity for carrier neogenesis cryptococcus, package both sexes it is mould
Plain B, to antimycotic and/or anti-leishmaniasis have good therapeutic effect, and preparation method it is easy to operate, without dirt
Dye.
Other advantages, target and feature of the invention will be illustrated in the following description to a certain extent, and
And to a certain extent, based on will be apparent to those skilled in the art to investigating hereafter, Huo Zheke
To be instructed from the practice of the present invention.Target of the invention and other advantages can be realized by following specification and
It obtains.
Detailed description of the invention
To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention is made below in conjunction with attached drawing excellent
The detailed description of choosing, in which:
Fig. 1 is the drug-polymer conjugate (PLGA-PEG- for having high-affinity to neogenesis cryptococcus prepared by example 1
AMB)1HNMR map；
Fig. 2 is the drug-polymer conjugate (PLGA-PEG- for having high-affinity to neogenesis cryptococcus prepared by example 1
AMB infared spectrum)；
Fig. 3 is the transmission electron microscope figure spectrum (TEM) for being loaded with the drug-polymer conjugate nanoparticle of amphotericin B；
Fig. 4 is the particle diameter distribution for the drug-polymer conjugate nanoparticle that laser particle analyzer detection is loaded with amphotericin B
Figure；
Fig. 5 is the nanoparticle and the external compatibility of neogenesis cryptococcus of Flow Cytometry Assay different surfaces ligand ratio,
Middle a, b, c are respectively the compatibility being incubated for after 0h, 1h and 2h.
Specific embodiment
Illustrate embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this specification
Other advantages and efficacy of the present invention can be easily understood for disclosed content.The present invention can also pass through in addition different specific realities
The mode of applying is embodied or practiced, the various details in this specification can also based on different viewpoints and application, without departing from
Various modifications or alterations are carried out under spirit of the invention.It should be noted that diagram provided in following embodiment is only to show
Meaning mode illustrates basic conception of the invention, and in the absence of conflict, the feature in following embodiment and embodiment can phase
Mutually combination.
Embodiment 1
Preparation has the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity to neogenesis cryptococcus, and method is such as
Under:
(1) 150.00mg (0.01mmol) poly(lactic-co-glycolic acid)-polyethylene glycol-N, hydroxysuccinimidyl acyl are weighed
Imines (is abbreviated as PLGA-PEG-NHS, wherein the degree of polymerization of poly lactide-glycolide acid is 50~80, polyethylene glycol
The degree of polymerization be 65~95), 9.25mg (0.01mmol) amphotericin B, be put into reaction flask, be added 4mL anhydrous DMF, then to
3.80mg (0.03mmol) N, N diisopropylethylamine (DIEA) is wherein added；
(2) reaction flask is placed on magnetic stirring apparatus, the temperature that magnetic stirring apparatus is arranged is 25 DEG C, is stirred to react for 24 hours；
(3) mixture after reaction is placed in the ice ether of 40mL, after placing 2h under the conditions of 4 DEG C, the speed of 4000rpm
It is centrifuged 5min, collects precipitated product, is washed repeatedly 3 times with ether, is dried in vacuo at 25 DEG C, obtaining light yellow solid is pair
Neogenesis cryptococcus has the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity.
Fig. 1 is the PLGA-PEG-AMB's prepared in example 11HNMR map.δ (ppm)=2.5 is deuterated DMSO signal
Peak, δ (ppm)=1.57 are that PLGA connects the methyl hydrogen (- CH in section3) signal peak, the peak of δ (ppm)=3.2 to 3.6 is sub- on PEG
The peak of methyl, δ (ppm)=4.81 are the signal peak (- CH of hydrogen in PLGA segment methylene2), δ (ppm)=5.12 is PLGA
In segment on tertiary base carbon hydrogen signal peak (- CH-), the signal peak of δ (ppm)=6.3 is H on polyenoid segment on amphotericin B20-
H30Signal peak shows that amphotericin B is connected on PLGA-PEG.
Fig. 2 is the infrared spectrogram of the PLGA-PEG-AMB prepared in example 1.3400cm-1Left and right characteristic absorption peak be
The stretching vibration peak of-OH, 2995cm on PLGA terminal hydroxyl and AMB-1And 2953cm-1Neighbouring absorption peak is saturation C-H bond
Stretching absorbance peak, 2881cm-1Neighbouring absorption peak is the stretching absorbance peak of methylene on the PEG of polymer one end, 848cm-1
It is-CH- in-plane bending deformation the absorption peak on PEG chain, 1759cm-1Peak be carbonyl in segment in PLGA stretching vibration absorb
Peak, 1099cm-1Wave band is the stretching absorbance peak of methylene on PEG, 1680cm- in PLGA-PEG-AMB1It is the flexible of-CO-NH-
Vibration peak shows that AMB is to connect in the form of covalent linkage with PLGA-PEG, rather than Electrostatic Absorption or other non-covalent
Key-shaped formula.
A kind of drug-polymer conjugate nanoparticle being loaded with amphotericin B, the preparation method is as follows:
(1) it will prepare in 15mg poly(lactic-co-glycolic acid)-polyethylene glycol (PLGA-PEG) and 5mg embodiment 1
Drug-polymer conjugate (PLGA-PEG-AMB) mixing, then 2mg amphotericin B is weighed, it is added in 1mL DMSO and stirs
Dissolution forms mixed solution；
(2) mixed solution is slowly injected into polyvinyl alcohol (0.5%, w/v) aqueous solution of (syringe needle 26G) 10mL, and surpassed
Sonication is to obtain oil-in-water (O/W) lotion；
(3) oil-in-water (O/W) lotion is subjected to 5000rpm and is centrifuged 5min, the biggish nanoparticle of particle separation diameter then will be upper
Layer aaerosol solution freezes (4 DEG C) centrifugation 30min by hypervelocity (14000rpm), collection nano particle, and with the ultrapure washing of 10mL
It washs and obtains sediment afterwards twice；
(4) obtained sediment is suspended in 5% aqueous sucrose solution, crosses 0.22 μm of filter membrane and removes the both sexes that do not wrap up
Mycin B (AMB) is freeze-dried to obtain the drug-polymer conjugate nanoparticle for being loaded with amphotericin B at -80 DEG C.
The drug-polymer conjugate nanoparticle transmission electron microscope figure spectrum for being loaded with amphotericin B is shown in Fig. 3, nanoparticle
Sublist face is smooth, spherical in shape, and partial size is about in 75nm or so；As shown in figure 4, laser particle analyzer measures nanoparticle partial size respectively about
Between 30~170nm, PDI is less than 0.3；PLGA-PEG-AMB should be mixed in a certain proportion with PLGA-PEG, be obtained to new
Raw cryptococcus has the nanoparticle of optimal affinity.
The nanoparticle of available different ratio when changing the additional proportion of PLGA-PEG and PLGA-PEG-AMB, will
PLGA-PEG and PLGA-PEG-AMB is respectively according to 1:0.1 (S1), 1:0.25 (S2), 1:0.5 (S3), 1:1.0 (S4), 1:2.0
(S5) what ratio was reacted is loaded with the drug-polymer conjugate nanoparticle of amphotericin B, contains respectively as material
FITC fluorescence probe is incubated for altogether after mixing the nanoparticle of the different ratio of preparation with fungi suspension respectively, takes in 0h, 1h and 2h
It uses one-stage water wash 3 times out, flow cytometer in 488nm measures the compatibility of nanoparticle and fungi as shown in figure 5, wherein a, b, c
Respectively it is incubated for the compatibility after 0h, 1h and 2h.The result shows that with the drug-polymer conjugate (PLGA-PEG- of addition
AMB ratio) increases (S1 to S5 ratio is incremented by), and the ligand density being prepared improves, and nanoparticle is affine with neogenesis cryptococcus
Property is also stronger.
Embodiment 2
Preparation has the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity to neogenesis cryptococcus, and method is such as
Under:
(1) 300.00mg (0.02mmol) PLGA-PEG-NHS (wherein polymerization of poly lactide-glycolide acid is weighed
Degree is 50~80, and the degree of polymerization of polyethylene glycol is 65~95), 9.25mg (0.01mmol) amphotericin B, be put into reaction flask
In, the anhydrous DMSO of 4mL is added, then 2.59mg (0.02mmol) N, N diisopropylethylamine (DIEA) is added thereto；
(2) reaction flask is placed on magnetic stirring apparatus, the temperature that magnetic stirring apparatus is arranged is 20 DEG C, is stirred to react 35h；
(3) mixture after reaction is placed in the ice ether of 40mL, after placing 2h under the conditions of 4 DEG C, the speed of 4000rpm
It is centrifuged 5min, collects precipitated product, is washed repeatedly 3 times with ether, is dried in vacuo at 25 DEG C, obtaining light yellow solid is pair
Neogenesis cryptococcus has the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity.
A kind of drug-polymer conjugate nanoparticle being loaded with amphotericin B, the preparation method is as follows:
(1) it will be prepared in 15mg poly(lactic-co-glycolic acid)-polyethylene glycol (PLGA-PEG) and 15mg embodiment 1
Drug-polymer conjugate (PLGA-PEG-AMB) mixing, then weigh 6mg amphotericin B, be added in 2mL DMSO and stir
It mixes dissolution and forms mixed solution；
(2) mixed solution is slowly injected into polyvinyl alcohol (1%, w/v) aqueous solution of (syringe needle 26G) 10mL, and ultrasound
Processing is to obtain oil-in-water (O/W) lotion；
(3) oil-in-water (O/W) lotion is subjected to 5000rpm and is centrifuged 5min, the biggish nanoparticle of particle separation diameter then will be upper
Layer aaerosol solution freezes (4 DEG C) centrifugation 30min by hypervelocity (14000rpm), collection nano particle, and with the ultrapure washing of 10mL
It washs and obtains sediment afterwards twice；
(4) obtained sediment is suspended in 3% aqueous sucrose solution, crosses 0.22 μm of filter membrane and removes the both sexes that do not wrap up
Mycin B (AMB) is freeze-dried to obtain the drug-polymer conjugate nanoparticle for being loaded with amphotericin B at -60 DEG C.
Embodiment 3
Preparation has the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity to neogenesis cryptococcus, and method is such as
Under:
(1) weighing 200.00mg (0.013mmol) PLGA-PEG-NHS, (wherein poly lactide-glycolide acid is poly-
Right is 50~80, and the degree of polymerization of polyethylene glycol is 65~95), 9.25mg (0.01mmol) amphotericin B, be put into reaction flask
In, 4mL anhydrous methylene chloride is added, then 5.18mg (0.04mmol) N, N diisopropylethylamine (DIEA) is added thereto；
(2) reaction flask is placed on magnetic stirring apparatus, the temperature that magnetic stirring apparatus is arranged is 30 DEG C, is stirred to react 12h；
(3) mixture after reaction is placed in the methanol of 40mL, after placing 2h under the conditions of 4 DEG C, the speed of 4000rpm from
Heart 5min collects precipitated product, is washed repeatedly 3 times with methanol, is dried in vacuo at 25 DEG C, and obtaining light yellow solid is to new
Raw cryptococcus has the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity.
A kind of drug-polymer conjugate nanoparticle being loaded with amphotericin B, the preparation method is as follows:
(1) it will prepare in 15mg poly(lactic-co-glycolic acid)-polyethylene glycol (PLGA-PEG) and 3mg embodiment 1
Drug-polymer conjugate (PLGA-PEG-AMB) mixing, then 1.2mg amphotericin B is weighed, it is added in 1mL DMSO and stirs
It mixes dissolution and forms mixed solution；
(2) mixed solution is slowly injected into polyvinyl alcohol (1.5%, w/v) aqueous solution of (syringe needle 26G) 10mL, and surpassed
Sonication is to obtain oil-in-water (O/W) lotion；
(3) oil-in-water (O/W) lotion is subjected to 5000rpm and is centrifuged 5min, the biggish nanoparticle of particle separation diameter then will be upper
Layer aaerosol solution freezes (4 DEG C) centrifugation 30min by hypervelocity (14000rpm), collection nano particle, and with the ultrapure washing of 10mL
It washs and obtains sediment afterwards twice；
(4) obtained sediment is suspended in 6% aqueous sucrose solution, crosses 0.22 μm of filter membrane and removes the both sexes that do not wrap up
Mycin B (AMB) is freeze-dried to obtain the drug-polymer conjugate nanoparticle for being loaded with amphotericin B at -100 DEG C.
Embodiment 4
Preparation has the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity to neogenesis cryptococcus, and method is such as
Under:
(1) weighing 200.00mg (0.013mmol) PLGA-PEG-NHS, (wherein poly lactide-glycolide acid is poly-
Right is 50~80, and the degree of polymerization of polyethylene glycol is 65~95), 9.25mg (0.01mmol) amphotericin B, be put into reaction flask
In, the anhydrous chloroform of 4mL is added, then 3.80mg (0.03mmol) N, N diisopropylethylamine (DIEA) is added thereto；
(2) reaction flask is placed on magnetic stirring apparatus, the temperature that magnetic stirring apparatus is arranged is 30 DEG C, is stirred to react 12h；
(3) mixture after reaction is placed in the isopropyl ether of 40mL, after placing 2h under the conditions of 4 DEG C, the speed of 4000rpm
It is centrifuged 5min, collects precipitated product, is washed repeatedly 3 times, is dried in vacuo at 25 DEG C, obtaining light yellow solid is with isopropyl ether
There is the drug-polymer conjugate (PLGA-PEG-AMB) of high-affinity to neogenesis cryptococcus.
In conclusion drug-polymer conjugate (PLGA-PEG-AMB) that the present invention is prepared and as carrier
The nanoparticle containing more amphotericin B of preparation all shows there is high compatibility to neogenesis cryptococcus, and it is mould to be loaded with both sexes
The polymer conjugates nanoparticle of plain B has good therapeutic effect to fungi and/or anti-leishmaniasis, and reason is:
A small amount of amphotericin B modification is copolymerized in poly lactic-co-glycolic acid in drug-polymer conjugate (PLGA-PEG-AMB)
Object-polyethylene glycol-N, the surface HOSu NHS (PLGA-PEG-NHS) have good compatibility to neogenesis cryptococcus；
And it uses drug-polymer conjugates (PLGA-PEG-AMB) to wrap up more amphotericin B as carrier and is formed after nanoparticle then
There can be good therapeutic effect to neogenesis cryptococcus, therefore can be even by the drug-polymer for being loaded with amphotericin B of preparation
Connection object nanoparticle is used to prepare the pharmaceutical preparation of antimycotic and/or anti-leishmaniasis, either as external medicine preparation,
Sucked medicine preparation or intravenous (IV) drug preparation can obtain good prevention or therapeutic effect.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to compared with
Good embodiment describes the invention in detail, those skilled in the art should understand that, it can be to skill of the invention
Art scheme is modified or replaced equivalently, and without departing from the objective and range of the technical program, should all be covered in the present invention
Scope of the claims in.
Claims (10)
1. the drug-polymer conjugate that a kind of pair of neogenesis cryptococcus has high-affinity, which is characterized in that the drug-polymerization
The general structure of object conjugate is as follows:
Wherein, the integer that m is 50~80, the integer that n is 65~95.
2. the preparation method of drug-polymer conjugate described in claim 1, which is characterized in that described the preparation method is as follows: will
Amphotericin B and poly(lactic-co-glycolic acid)-polyethylene glycol-N, HOSu NHS are added to containing N, and N- bis- is different
In the anhydrous organic solvent of propylethylamine, coupling reaction is carried out, precipitating reagent is added after reaction, solid reaction is obtained by filtration
Object, 20~30 DEG C at a temperature of carry out vacuum drying the drug-polymer for having high-affinity to neogenesis cryptococcus can be obtained
Conjugate.
3. preparation method according to claim 2, which is characterized in that the amphotericin B and poly lactic-co-glycolic acid are total
Polymers-polyethylene glycol-N, the molar ratio of HOSu NHS are 1:1~2:1, and the poly lactide-glycolide acid-is poly-
Ethylene glycol-N, the degree of polymerization of poly lactide-glycolide acid is 50~80 in HOSu NHS, the polymerization of polyethylene glycol
Degree is 65~95.
4. preparation method according to claim 2, which is characterized in that the organic solvent is DMF, DMSO, methylene chloride
Or any one in chloroform, in the anhydrous organic solvent concentration containing n,N-diisopropylethylamine be 5.0~
10.0mmol/L。
5. preparation method according to claim 2, which is characterized in that the temperature of the coupling reaction is 20~30 DEG C, instead
It is 12~35h between seasonable.
6. preparation method according to claim 2, which is characterized in that the precipitating reagent is ether, isopropyl ether or methanol
In any one.
7. a kind of drug-polymer conjugate nanoparticle for being loaded with amphotericin B, which is characterized in that the nanoparticle includes power
Benefit requires the 1 drug-polymer conjugate and amphotericin B, and the amphotericin B is by the drug-polymer conjugate
Package.
8. a kind of preparation method for the drug-polymer conjugate nanoparticle for being loaded with amphotericin B described in claim 7, special
Sign is that the preparation method includes the following steps:
(1) by poly(lactic-co-glycolic acid)-polyethylene glycol and described there is the drug-of high-affinity poly- neogenesis cryptococcus
It closes object conjugate and according to 1~5:1 is mixed to form mixture, by the mixture and amphotericin B according to the mass ratio of 5~15:1
Mixing is added DMSO and dissolves to form mixed solution；
(2) mixed solution is slowly injected into polyvinyl alcohol water solution, and be ultrasonically treated to obtain oil-in-water emulsion；
(3) oil-in-water emulsion is centrifuged, upper layer aaerosol solution is taken to be centrifuged by Superfreezing, collected nano particle, be used in combination
Milli-Q water obtains sediment；
(4) obtained sediment is suspended in 3~6% aqueous sucrose solutions, excessively 0.22 μm of filter membrane, it is cold at -60 DEG C~-100 DEG C
The dry drug-polymer conjugate nanoparticle that can be obtained and be loaded with amphotericin B is lyophilized.
9. application of the nanoparticle described in claim 7 in the pharmaceutical preparation for preparing antimycotic and/or anti-leishmaniasis.
10. application according to claim 9, which is characterized in that the pharmaceutical preparation includes external medicine preparation, sucking medicine
Any one in object preparation or intravenous (IV) drug preparation.
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