CN107141323B - Reduction/pH dual responsiveness adriamycin prodrug and the preparation method and application thereof - Google Patents
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Abstract
The invention discloses a kind of reduction/pH dual responsiveness adriamycin prodrugs and the preparation method and application thereof.Specific preparation method is 3,3 '-two sulphur diine butyl dipropionate (B- of small molecule compound blocked using alkynylss- B) and azido sealing end two azidoethyl of compound, two acetal radical polyethylene glycol (N3‑a‑PEG‑a‑N3) chemically reacted, prepare alkynyl sealing end has reduction/pH dual responsiveness polymer poly (SS-alt‑A)n；Then doxorubicin derivative is connected to the both ends of polymer segment, obtain reduction/pH dual responsiveness adriamycin prodrug DOX-hyd‑poly(SS‑alt‑A)n‑hyd-DOX.This kind of characteristic with the water-soluble adriamycin prodrug of reduction/pH dual responsiveness with good biocompatibility and controllable in drug release, thus may be used as stimulation sensibility anti-tumor predrug.
Description
Technical field
The invention belongs to field of biomedical polymer materials, and in particular to a kind of reduction/pH based on polyethylene glycol is bis-
Weight responsiveness adriamycin prodrug, preparation method and its application.
Background technique
Cancer is body in many factors, multiple stages and is repeatedly mutated and leads to normal cell canceration as a result, curing
Malignant tumour is also referred to as on.The lethality of cancer is big, easily shifts, by destroying the various functions of tissue and organ, finally
People are caused to die because of dying of exhaustion for organ dysfunction.According to statistics, with the growth of population, the disease incidence of cancer is also constantly increasing
It is long, the health of the mankind is seriously endangered, therefore, attention of the cancer treatment method by domestic and international medical investigators.
In in the past few decades, with the progress of medical technology, the treatment method of various cancers is rapidly developed,
Including operative treatment, chemotherapy, radiotherapy, immunization therapy, hormone therapy, targeted therapy etc..But make in chemotherapy process
Small-molecule drug (such as: taxol, adriamycin, camptothecine etc.), to normal cell while killing tumour cell effect
Also there is very strong damaging action, therefore, the side effects such as serious vomiting, dizzy, the decline of body function can be caused.Meanwhile small point
Sub- drug is easily identified and is discharged by body during blood circulation, in addition environment special near tumor tissues, such as crisscross multiple
Miscellaneous blood vessel network structure, fine and close interstitial structure and higher interstitial fluid pressure etc., so as to be hard to reach tumour thin for drug
It is intracellular.The curative effect of anti-tumor drug clinically is affected just because of these restrictive conditions.
By prodrug, the above problem can be efficiently solved.Prodrug (prodrug), also referred to as prodrug, premedicant
Object refers to the compound after organism conversion with pharmacological action.Polymeric prodrugs refer to active drug and rise
The polymer support of translocation makes drug not have activity by Covalent bonding together.But when prodrug enter body circulation and
In metabolic process, active medicine can be released by hydrolysis removal carrier, play pharmacological action.This mode can have
Effect ground improves the utilization rate of drug, enhances targeting, reduces the toxic side effect of drug.
Most important for the selection of polymeric base material in polymeric prodrugs structure, biocompatibility is material
Most basic requirement for pharmaceutical carrier.Polyethylene glycol (PEG) has non-toxic, good biocompatibility and biodegradable
Property, ratified to use by U.S. Food and Drug Administration (FDA).By to PEG carry out end-functionalization modification, then with dredge
Aqueous pharmaceutical bonding, can prepare polymeric prodrugs, assign some advantageous properties of drug molecule.Although polymeric prodrugs are in cancer
Have how more drug is still transmitted in tumour cell by very big advantage in treating, is still to have been a concern
Problem.Drug reaches lesion site and needs to overcome various barriers, is considered as how keeping transporting carrier during blood circulation
Stability, extend circulation time；How efficiently to enter into the cell, more accumulates at specific position；How to drug
Controlled release etc..If do not solved these problems, drug will be greatly reduced to the therapeutic effect of cancer.
In the prior art, have some about the sensitive report with sensitivity to acid prodrug of reduction.But as before antitumor
Medicine should have good biocompatibility and biodegradability, also, as anti-tumor predrug, should also have following
Feature: stable polymer micelle can be formed in aqueous solution, hydrophily shell plays stable micella, raising micella blood follows
The effect of ring time；When prodrug micelle recycles in vivo, the performance with anticoagulation and anti-protein adsorption；When carrier micelle reaches
When tumour or pathological tissues, can utilize the low pH condition of local organization and high glutathione concentrations the characteristics of, destroy micella,
Quick release goes out anticancer drug.Therefore, it is necessary to seek more drug effects it is obvious and have in tumour cell microenvironment reduction and
Sensitivity to acid anti-tumor predrug.
Summary of the invention
The reduction based on polyethylene glycol that the object of the present invention is to provide a kind of/pH dual responsiveness adriamycin prodrug and its system
Preparation Method；The adriamycin prodrug has good biocompatibility and inhibits the ability of tumor cell proliferation.
The specific technical solution of the present invention are as follows: a kind of reduction/pH dual responsiveness adriamycin prodrug, by the following chemical structure
Formula expression:
M is that 3~113, n is 3~15.
In above-mentioned technical proposal, adriamycin prodrug has the group of sensitivity to acid and the group of reduction-sensitive；The tool
The hydrophobicity adriamycin part for having the adriamycin prodrug of dual responsiveness to contain in structure, is used to form the kernel of micella, PEG
Hydrophilic parts are wrapped in the shell that micella is formed outside kernel, have good effect to the stability of micella.Reduction and
Under the conditions of pH, acid sensitive group and reduction sensitive group are broken, and micella is destroyed, and is gathered in rapidly release
Hydrophobic anticancer drug inside micella.
In preferred technical solution, reduction/pH dual responsiveness adriamycin prodrug number-average molecular weight is 4000~80000
g·mol-1。
The present invention is using the alternate copolymer poly (SS- in structure containing acid sensitive group and reduction sensitive groupalt- A) it is used as base stock, it is reacted after mantoquita and ligand catalysis chemical reaction activation with anticancer drugs, doxorubicin, prepares base
In reduction/pH dual responsiveness adriamycin prodrug DOX- of polyethylene glycolhyd-poly(SS-alt-A)-hyd-DOX。
Above-mentioned reduction/pH dual responsiveness adriamycin prodrug preparation method, comprising the following steps:
(1) using 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride (DOX × HCl) as raw material, anhydrous methanol is solvent, ice
Acetic acid is catalyst, obtains doxorubicin derivative by chemical reaction；
Wherein, the molar ratio of 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride is 1:(2~6)；
The chemical structural formula of the 6- nitrine hexanoyl hydrazine are as follows:
The chemical structural formula of the doxorubicin derivative are as follows:
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and ligand, with 3,3 '-two sulphur diine butyl
Dipropionate and two azidoethyls, two acetal radical polyethylene glycol are that raw material passes through using anhydrous n,N-Dimethylformamide as solvent
Reaction prepares the polyethylene glycol alternate copolymer of two end alkynyl radicals sealing end；
Wherein, two azidoethyls, two acetal radical polyethylene glycol, 3,3 '-two sulphur diine butyl dipropionates and copper salt catalyst
Molar ratio be 1: (1.1~1.5): (0.5~1.5)；Copper salt catalyst and the molar ratio of ligand are 1: (1~2)；
The chemical structural formula of 3,3 '-two sulphur diine butyl dipropionate are as follows:
The structural formula of two azidoethyls, the two acetal radical polyethylene glycol are as follows:
Above-mentioned m is 3~113；
The structural formula of the polyethylene glycol alternate copolymer of the two end alkynyl radicals sealing end are as follows:
Above-mentioned m is that 3~113, n is 3~15.
(3) using anhydrous n,N-Dimethylformamide as solvent, in the presence of copper salt catalyst and ligand, step is used
(2) the polyethylene glycol alternate copolymer of the two end alkynyl radicals sealing end obtained is reacted with the doxorubicin derivative that step (1) obtains, and is made
It is standby to obtain the reduction/pH dual responsiveness adriamycin prodrug；
The polyethylene glycol alternate copolymer of the two end alkynyl radicals sealing end, the molar ratio of doxorubicin derivative are 1:(2~5).
In above-mentioned technical proposal, doxorubicin derivative (N is prepared3-hyd- DOX × HCl), it is main to pass through three step preparations
It forms:
The first step prepares 6- nitrine methyl caproate.With 6- bromocaproic acid methyl esters and sodium azide (NaN3) reaction, it is with DMF
Solvent carries out azido reaction, obtains 6- nitrine methyl caproate.Wherein, 6- bromocaproic acid methyl esters, sodium azide molar ratio be 1:
(1.5~5)；
Second step prepares 6- nitrine hexanoyl hydrazine.It is 85% with the 6- nitrine methyl caproate and mass fraction of first step synthesis
It is hydrated hydrazine reaction, using tetrahydrofuran (THF) as solvent, by amidation process, obtains 6- nitrine hexanoyl hydrazine.Wherein, 6- is folded
The molar ratio of nitrogen methyl caproate and hydrazine hydrate is 1:(10~30)；
Third step prepares doxorubicin derivative (N3-hyd- DOX × HCl).The 6- nitrine hexanoyl hydrazine synthesized using second step
It is used as raw material with doxorubicin hydrochloride (DOX × HCl), using anhydrous methanol as solvent, glacial acetic acid is catalyst, obtains adriamycin and spreads out
Biology.Wherein, the molar ratio of 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride is 1:(2~6)；
The polyethylene glycol alternate copolymer for preparing two end alkynyl radicals sealing end includes the following steps:
Prepare 3,3 '-two sulphur diine butyl dipropionate (B- of small molecule compound alkynyl sealing end and containing disulfide bondss- B).It is raw material using 3,3 '-dithiodipropionic acids and 3- butyne-1-ol, with 1- ethyl-(3- dimethylaminopropyl) carbon
Diimmonium salt hydrochlorate is activator, and 4-dimethylaminopyridine is catalyst, and methylene chloride is solvent, by esterification, is obtained
3,3 '-two sulphur diine butyl dipropionates.Wherein, 3, the molar ratio of 3 '-dithiodipropionic acids and 3- butyne-1-ol is 1:(1.5
~3)；
Prepare two azidoethyls, the two acetal radical polyethylene glycol (N of azido sealing end3-a-PEG-a-N3), pass through two steppings
Row:
The first step synthesizes two acetal radical polyethylene glycol (Cl- of Dichloroethyla-PEG-a- Cl).Using polyethylene glycol (HO-
It PEG-OH is) raw material with 2- chloroethyl vinyl ether (CEVE), in the presence of acid catalyst, reaction obtains the poly- second that end is chloro
Diol product Cl-a-PEG-a-Cl.Wherein, the molecular weight of polyethylene glycol are as follows: 400 ~ 5000 gmol-1, polyethylene glycol and 2- chlorine
The molar ratio of ethyl vinyl ether is 1:(2~6)；Acid catalyst is p-methyl benzenesulfonic acid, p-methyl benzenesulfonic acid pyridine, trifluoro second
One of acid；
Second step synthesizes two azidoethyls, two acetal radical polyethylene glycol (N3-a-PEG-a-N3).The Cl- that the first step is obtaineda-PEG-a- Cl and reaction of sodium azide obtain two azidoethyls, two acetal radical polyethylene glycol (N using methylene chloride as solvent3-a-PEG-a-N3).Wherein, polyethylene glycol (Cl- of the end containing chloroa-PEG-a- Cl) with the molar ratio of sodium azide be
1:(2~6)；
Prepare the polymer poly (SS- of alkynyl sealing endalt-A).Under nitrogen protection, in copper salt catalyst and ligand
In the presence of, utilize the 3 of above-mentioned synthesis, 3 '-two sulphur diine butyl dipropionate (B-ss- B) it is poly- with two azidoethyls, two acetal radical
Ethylene glycol (N3-a-PEG-a-N3) chemically react, prepare the polymer poly (SS- of alkynyl sealing endalt-A).Wherein, two is folded
Two acetal radical polyethylene glycol (N of nitrogen ethyl3-a-PEG-a-N3) and 3,3 '-two sulphur diine butyl dipropionate (B-ss- B) mole
Than for 1:(1.1~1.5)；
Prepare the adriamycin prodrug DOX- of double-responsehyd-poly(SS-alt-A)-hyd- DOX includes the following steps:
Utilize the polyethylene glycol alternating copolymerization for the two end alkynyl radicals sealing end that the doxorubicin derivative of desalting processing is synthesized with upper step
Object poly (SS-alt- A) it chemically reacts, obtain reduction/pH dual responsiveness adriamycin prodrug DOX-hyd-poly(SS-alt-A)-hyd-DOX.Wherein, the polyethylene glycol alternate copolymer of two end alkynyl radicals sealing end and the molar ratio of doxorubicin derivative are
1:(2~5).
In above-mentioned technical proposal, in step (1), reaction temperature is 60 DEG C~80 DEG C, and the reaction time is the h of 12 h~24；Step
Suddenly in (2), inert gas is nitrogen；Reaction temperature is 25 DEG C~50 DEG C, and the reaction time is the h of 24 h~72；In step (3), instead
Answering temperature is 25 DEG C~50 DEG C, and the reaction time is the h of 24 h~48.
In above-mentioned technical proposal, in step (2) and step (3), copper salt catalyst is selected from Salzburg vitriol, stannous chloride
Or cuprous bromide；Ligand is selected from: sodium ascorbate, bipyridyl, five methyl diethylentriamine, tetramethylethylenediamine or hexamethyl
One of trien.
The present invention in the presence of copper salt catalyst and ligand, by the restriction of raw material and parameter, is prepared for the first time
Polyethylene glycol alternate copolymer containing acid sensitive group and reduction sensitive group, solving existing reaction system can not prepare
Out simultaneously containing acid sensitive group and the problem of restore the alternate copolymer of sensitive group, be it is a kind of it is novel, simple, efficient,
Quick synthetic method.
Further technical solution carries out purification processes, the purifying to product respectively after the completion of step (1)~(3)
Process the following steps are included:
(i) doxorubicin derivative N3-hyd- DOX × HCl purifying: after reaction, reaction solution is filtered to remove with cotton
Sediment precipitates three times in ice ether after removing solvent；Sediment is obtained using centrifugal process, places it in vacuum oven
In be dried to obtain the product N of red powder3-hyd-DOX×HCl；
(ii) the polyethylene glycol alternate copolymer poly (SS- of two end alkynyl radicals sealing endalt- A) purifying: after reaction,
After 48 h of DMF dialysis, it is placed in 60 DEG C of oil baths, DMF is removed under reduced pressure；Then with 150 mL CH2Cl2It re-dissolves, with saturation
NaCl aqueous solution is extracted, and mantoquita is removed；Organic layer anhydrous Na2SO4After dry 4 h, rotary evaporation removing solvent, then
It is precipitated 3 times in diethyl ether solution, sediment is collected and is dried in vacuo, finally obtain sticky product poly (SS-alt-A)；
(III) reduction/pH dual responsiveness adriamycin prodrug DOX-hyd-poly(SS-alt-A)-hydThe purifying of-DOX:
After reaction, using 72 h of deionized water dialysis, red, transparent liquid freezing obtained in bag filter is dry, it obtains deep
Red solid product DOX-hyd-poly(SS-alt-A)-hyd-DOX。
In above-mentioned technical proposal, in step (i), solvent is removed using the method for rotary evaporation；In step (ii) when dialysis
Use molecular cut off for the bag filter of 1000~14000 Da；In step (III), when dialysis, uses molecular cut off for 3500
The bag filter of~14000 Da.
Specific purification the following steps are included:
(i) doxorubicin derivative (N is prepared3-hyd- DOX × HCl) purifying:
The first step prepares the purifying of 6- nitrine methyl caproate: after reaction, being filtered with the short column of neutral aluminum oxide
Reaction solution, and solvent DMF is removed under reduced pressure with oil pump.Then, crude product is dissolved with a large amount of methylene chloride, it is secondary with 30 mL
Water extracts three times, and organic phase is collected and dries 4 h with anhydrous sodium sulfate.Finally, being removed with cotton filtering, rotary evaporation molten
Agent, place the product in 24 h dry in vacuum drying oven, obtain colourless product liquid 6- nitrine methyl caproate；
Second step prepares the purifying of 6- nitrine hexanoyl hydrazine: after reaction, rotary evaporation uses CH instead after removing THF2Cl2It is molten
Solution.In separatory funnel, crude product is extracted using saturated sodium-chloride water solution.By organic phase anhydrous Na2SO4Dry 4 h, are removed
It is dried in vacuo after removing solvent, is finally collected into product 6- nitrine hexanoyl hydrazine；
Third step prepares the purifying of doxorubicin derivative: after reaction, being filtered to remove in reaction solution with cotton
Na2SO4, after filtrate rotary evaporation is removed solvent, precipitated three times in ice ether.Finally, following sink is obtained using centrifugal process
Starch places it in dry 24 h in vacuum oven, obtains red powder product, as N3-hyd-DOX×HCl；
(ii) 3,3 '-two sulphur diine butyl dipropionate (B- of alkynyl sealing end are preparedss- B) purifying: reaction terminate
Afterwards, it is filtered with filter paper, except the salt generated in dereaction, then with the CH of 150 mL2Cl2Dissolution, with 1 M HCl solution and saturation chlorine
The mixed solution for changing sodium (NaCl) aqueous solution extracts 2 times, and each water layer continues the CH with 30 mL2Cl2Extraction 3 times.By organic layer
After collection, anhydrous Na is used2SO4Dry 4 h, the crude product after removing solvent continue to carry out separating-purifying using column chromatography.It collects
Final product is dried in vacuo 24 h to get to 3,3 '-two sulphur diine butyl dipropionate (B-ss- B)；
(III) prepares two azidoethyls, the two acetal radical polyethylene glycol (N of azido sealing end3-a-PEG-a-N3) purifying:
The first step, preparation synthesis two acetal radical polyethylene glycol (Cl- of Dichloroethyla-PEG-a- Cl) purifying: reaction terminate
Afterwards, CH is first used2Cl2Dilute reaction solution, with the phosphate buffer solution (PBS, pH 10.0) of 10 mL and mixing for saturation NaCl aqueous solution
Solution extraction is closed, obtained water layer continues to use CH2Cl2Extraction is three times.Organic layer anhydrous Na2SO4After dry 4 h, rotary evaporation
A large amount of solvents are removed, when remaining about 3 mL crude product in solution, it are precipitated in n-hexane three times, sediment is collected and vacuum is dry
It is dry, obtain product Cl-a-PEG-a-Cl；
Second step, preparation synthesis two azidoethyls, two acetal radical polyethylene glycol (N3-a-PEG-a-N3) purifying: reaction knot
Shu Hou, with neutral Al2O3Short column be filtered to remove unreacted NaN3, solvent DMF is removed under 60 °C of oil bath reduced pressures.With
Afterwards, CH is used2Cl2Crude product is dissolved, with the PB buffer solution purification by liquid extraction product of pH 10.0, each water layer continues with 20 mL's
CH2Cl2Extraction, organic phase is finally collected and use anhydrous Na2SO4Dry, rotary evaporation, which removes solvent and is dried in vacuo, obtains palm fibre
The product N of color3-a-PEG-a-N3；
The polyethylene glycol alternate copolymer poly (SS- of (IV) two end alkynyl radical sealing endalt- A) purifying: after reaction,
After 48 h of DMF dialysis, it is placed in 60 DEG C of oil baths, DMF is removed under reduced pressure.Then with 150 mL CH2Cl2It re-dissolves, with saturation
NaCl aqueous solution is extracted, and mantoquita is removed.Organic layer anhydrous Na2SO4After dry 4 h, rotary evaporation removing solvent, then
It is precipitated 3 times in ether, sediment is collected and is dried in vacuo, finally obtain sticky product poly (SS-alt-A)；
(V) prepares the adriamycin prodrug DOX- of double-responsehyd-poly(SS-alt-A)-hydThe purifying of-DOX: anti-
It is using 72 h of deionized water dialysis, red, transparent liquid freezing obtained in bag filter is dry after answering, obtain red
Sticky solid material DOX-hyd-poly(SS-alt-A)-hyd-DOX。
Reduction disclosed by the invention/pH dual responsiveness adriamycin prodrug DOX-hyd-poly(SS-alt-A)-hyd-DOX
It can be self-assembly of prodrug micelle in aqueous solution, hydrophobic drug adriamycin forms the core of micella；Hydrophily PEG chain section shape
At the shell of micella, play the role of stable micella.And contain reduction response between hydrophobic cores and hydrophily PEG chain section
The acetal and acylhydrazone key group of disulfide bond and acid-sensitive, under the conditions of reduction and pH, sensitive group is broken, and micella is broken
It is bad, to rapidly release the hydrophobic anticancer drug being gathered in inside micella.Therefore, the present invention is claimed above-mentioned simultaneously
Reduction/pH dual responsiveness adriamycin prodrug is preparing the application in stimulating responsive anticancer nano drug.
Due to the implementation of above scheme, compared with prior art, the present invention having the advantage that
1, the present invention is using the good polyethylene glycol of biocompatibility and hydrophobic anticancer drug adriamycin respectively as parent
Aqueous segment and hydrophobic end, to be connected under the reducing conditions with the group that can be hydrolyzed under acid condition as sensibility
Group is prepared for the reduction with good biocompatibility/pH dual responsiveness adriamycin prodrug；Compared with raw medicine adriamycin,
Prodrug has good water-soluble and storage stability, easily decomposes in tumour cell environment, accelerates drug release, increases
Its application value is added.
2, reduction/pH dual responsiveness adriamycin prodrug that the present invention obtains can be self-assembly of stable polymerization in water
Object prodrug micelle.Hydrophobic adriamycin moiety aggregation forms micelle inner core, and hydrophily PEG chain section plays surely as micella shell
The effect determined micella, improve micella blood circulation time.Hydrophilic segment and hydrophobic part pass through cleavable in acid condition
Key is learned to be connected, at the same inside hydrophilic PEG chain segment containing under the reducing conditions with the group that can be hydrolyzed under acid condition；When
When prodrug micelle reaches tumour or pathological tissues, since the pH value of these tissue microenvironments reduces, acid sensitive group can be made
Fracture, while in the environment of high glutathione, reduction sensitive group is also broken, and cause micellar structure to be destroyed, from
And quick release goes out anticancer drug, increases the utilization rate and targeting of drug, in terms of the treatment of cancer, has and potentially answers
With value.
3, reduction provided by the invention/pH dual responsiveness adriamycin pro-drugs are clear, and synthesis condition is not harsh, has
Following distinguishing feature: (1) raw materials used and reagent is easy to get；(2) reaction condition is simple；(3) yield is high；(4) three-dimensional
Selectivity is good；(5) product separation is easy；(6) product stability is good；Preparation is easy, and purification is convenient, is suitble to industrialized production.
Detailed description of the invention
Fig. 1 is doxorubicin derivative (N in embodiment one3-hyd- DOX × HCl) hydrogen nuclear magnetic resonance spectrogram, solvent is deuterium
For dimethyl sulfoxide；
Fig. 2 is the hydrogen nuclear magnetic resonance spectrogram of 3,3 '-two sulphur diine butyl dipropionates in embodiment two, and solvent is deuterated chlorine
It is imitative；
Fig. 3 is Cl- in embodiment threea-PEG21-a- Cl and N3-a-PEG21-a-N3Hydrogen nuclear magnetic resonance spectrogram, solvent is
Deuterated chloroform, wherein 21 representative-CH of subscript2CH2The number of repeat unit of O-；
Fig. 4 is polymer poly (SS- in example IValt-A)3.6Hydrogen nuclear magnetic resonance spectrogram, solvent is deuterated chloroform,
Wherein, subscript 3.6 represents the degree of polymerization of alternate copolymer；
Fig. 5 is DOX- in embodiment fivehyd-poly(SS-alt-A)3.6-hydThe hydrogen nuclear magnetic resonance spectrogram of-DOX, solvent
For deuterated chloroform；
Fig. 6 is HO-PEG in embodiment one~five21-OH, Cl-a-PEG21-a-Cl, N3-a-PEG21-a-N3, poly
(SS-alt-A)3.6, N3-hyd- DOX × HCl and DOX-hyd-poly(SS-alt-A)3.6-hydThe infrared spectrogram of-DOX；
Fig. 7 is DOX- in embodiment fivehyd-poly(SS-alt-A)3.6-hyd- DOX in 7.4 buffer solution of pH from
Assemble the dynamic light scattering curve and transmission electron microscope photo figure of the micella formed；
Fig. 8 is DOX- in embodiment fivehyd-poly(SS-alt-A)3.6-hyd- DOX formed polymeric prodrugs micella with
Drug release patterns figure of the naked medicine adriamycin in the buffer solution of different pH value；
Fig. 9 is DOX-hyd-poly(SS-alt-A)3.6-hydThe inhibition tumour cell of-DOX prodrug and naked medicine adriamycin increases
Grow performance test figure.
Specific embodiment
Below with reference to embodiment and attached drawing, the invention will be further described:
Embodiment one: doxorubicin derivative (N3-hyd- DOX × HCl) synthesis
Doxorubicin derivative (N3-hyd- DOX × HCl) mainly it is prepared by three steps.The first step, preparation 6- are folded
Nitrogen methyl caproate；Second step prepares 6- nitrine hexanoyl hydrazine；Third step modify to doxorubicin hydrochloride (DOX × HCl)
To doxorubicin derivative (N3-hyd- DOX × HCl), specific synthetic method is as follows:
Synthesized micromolecule compound 6- nitrine methyl caproate.The device for being placed in 120 °C of baking ovens in advance is taken out, is put into
It in desiccator, is cooled to room temperature to it, takes out and use.6- bromocaproic acid methyl esters (3.54 g, 16.93 mmol) is weighed respectively and is folded
Sodium nitride (NaN3) (2.75 g, 42.33 mmol), and be dissolved in the round-bottomed flask of 50 mL with the DMF of 20 mL, at 60 °C
Terminate after reacting 12 h under the conditions of condensate return.With neutral aluminum oxide (Al2O3) short column filtering reacting liquid, remove not
The NaN of reaction3, and solvent DMF is removed under reduced pressure with oil pump.Then, with a large amount of methylene chloride (CH2Cl2) dissolution crude product, it uses
The secondary water extraction of 30 mL three times, organic phase is collected and uses anhydrous sodium sulfate (Na2SO4) 4 h of drying.Finally, with cotton mistake
Filter, rotary evaporation remove solvent, and place the product in 24 h dry in vacuum drying oven, obtain colourless product liquid 6- nitrine caproic acid
Methyl esters (2.47 g, yield: 53.2%)；
Obtained 6- nitrine methyl caproate (0.76 g, 4.44 mmol) is added in round-bottomed flask and mass fraction is
85% hydrazine hydrate (5.60 g, 111.1 mmol) is used as solvent using tetrahydrofuran (THF).Equally, it in 80 °C of oil baths and fills
Have in the device of serpentine condenser, 12 h of back flow reaction.After reaction, rotary evaporation removes THF, uses CH2Cl2Dissolution.Dividing
In liquid funnel, crude product is extracted using saturated sodium-chloride water solution.By organic phase anhydrous Na2SO4Dry 4 h, after removing solvent
Vacuum drying, is finally collected into product 6- nitrine hexanoyl hydrazine (0.55 g, yield: 56.2%)；
6- nitrine hexanoyl hydrazine (102.0 mg, 0.60 mmol), DOX × HCl(115.9 mg, 0.20 mmol are weighed respectively)
And anhydrous Na2SO4(103 mg, 0.73 mmol) is added in the round-bottomed flask with condensation reflux unit, with 30 mL without water beetle
Alcohol dissolution.3 drop glacial acetic acid are then added, after stirring, are protected from light 24 h in 65 °C of oil baths.After reaction,
It is filtered with cotton, removes the Na in reaction solution2SO4, precipitated three times in ice ether after removing solvent.Finally, it is obtained using centrifugal process
To following sediment, dry 24 h in vacuum oven are placed it in, the product of red powder, as N are obtained3-hyd- DOX × HCl(93.5 mg, yield: 62.4%).
Embodiment two: 3,3 '-two sulphur diine butyl dipropionate (B- of alkynyl sealing endss- B) synthesis
Firstly, 3 '-dithiodipropionic acids and 3- butyne-1-ol are raw material, with 3 with 1- ethyl-(3- dimethylamino third
Base) carbodiimide hydrochloride is activator, 4-dimethylaminopyridine is catalyst, and methylene chloride is solvent, by esterification,
Obtain 3,3 '-two sulphur diine butyl dipropionates.Specific synthetic method is as follows:
Glass apparatus used is placed in 120 °C of baking ovens and dries 4 h, taking-up, which is placed in desiccator, to be cooled to room temperature for use.
Firstly, sequentially adding 3,3 '-dithiodipropionic acids (3.66 g, 17.40 mmol), 1- ethyl-(3- diformazan into three-neck flask
Base aminopropyl) carbodiimide hydrochloride (EDC × HCl) (8.67 g, 45.23 mmol) and 4-dimethylaminopyridine (DMAP)
(3.19 g, 2.61 mmol), with the CH of 100 mL2Cl2Dissolution；Secondly, by 3- butyne-1-ol (3.17 g, 45.23 mmol)
With the CH of 15 mL2Cl2It is added in constant pressure funnel.2 h are added dropwise in ice-water bath, after completion of dropwise addition, device is moved on to 25 °C
Oil bath in, the reaction was continued 48 h；
After reaction, rotary evaporation removes a small amount of solvent, and filter paper is filtered to remove the salt that reaction generates, then with 150 mL's
CH2Cl2Dissolution is extracted 2 times, the water layer of collection with the mixed solution of 1 M HCl solution and saturated sodium-chloride (NaCl) aqueous solution
With the CH of 30 mL2Cl2Extraction 3 times.After organic layer is collected, anhydrous Na is used2SO4Dry 4 h, crude product after removing solvent after
Continuous to carry out separating-purifying using column chromatography, eluent is the mixed solution (ethyl acetate: petroleum ether of ethyl acetate and petroleum ether
=1:6).Final product is collected, and obtains purer 3,3 '-two sulphur diine butyl dipropionate B- after being dried in vacuo 24 hss-
B(3.63 g, yield: 59.4%).
Embodiment three: two azidoethyls, the two acetal radical polyethylene glycol (N of azido sealing end3-a-PEG21-a-N3) synthesis
Two azidoethyl of compound, the two acetal radical polyethylene glycol (N of azido sealing end3-a-PEG21-a-N3), pass through two steps
Reaction obtains: firstly, synthesis two acetal radical polyethylene glycol (Cl- of Dichloroethyla-PEG21-a- Cl)；Secondly, two nitrine second of synthesis
Two acetal radical polyethylene glycol (N of base3-a-PEG21-a-N3).Specific synthetic method is as follows:
The first step synthesizes two acetal radical polyethylene glycol (Cl- of Dichloroethyla-PEG21-a- Cl).Taking molecular weight is 1000
Polyethylene glycol (HO-PEG22- OH) and (6.01 g, 6.01 mmol) and p-methyl benzenesulfonic acid pyridine (PPTS) (0.30 g, 1.20
Mmol it) is placed in the side-neck flask of 100 mL, and the toluene of 50 mL is added, removed using the method for two sub-atmospheric pressure toluene azeotropic
Water impurity.After its cooling, under nitrogen protection, the dry CH of 30 mL is added2Cl2Dissolution, and add in constant pressure funnel
Enter the CH of 10 mL2Cl2With 2- chloroethyl vinyl ether (CEVE) (3.20 g, 30.05 mmol), after ice-water bath drips, room temperature
Lower the reaction was continued 1 h, is added the sodium carbonate (Na that the mass fraction of 10 mL is 5%2CO3) aqueous solution termination reaction；
The method that the purification of product is combined using extraction and precipitated phase.First use CH2Cl2Dilute reaction solution, with 10 mL phosphoric acid
Buffer solution (PBS, pH 10.0) and the mixed solution extraction for being saturated NaCl aqueous solution, obtained water layer continue to use CH2Cl2Extraction
Three times.Organic layer anhydrous Na2SO4After dry 4 h, rotary evaporation removes CH2Cl2Solvent.Then, it is sunk in n-hexane
It forms sediment three times, collect sediment and is dried in vacuo, obtain product Cl-a-PEG21-a- Cl(5.75g, yield: 78.9%)；
Second step synthesizes two azidoethyls, two acetal radical polyethylene glycol (N3-a-PEG21-a-N3).The Cl- that upper step is obtaineda-PEG-a- Cl(2.37 g, 1.95 mmol) and NaN3(0.76 g, 11.70 mmol) are dissolved in the DMF of 25 mL, 60 °C
48 h are reacted under oil bath.After reaction, with neutral Al2O3Short column be filtered to remove unreacted NaN3, it is placed in 60 DEG C of oil baths
In, DMF is removed under reduced pressure.Then, CH is used2Cl2Crude product is dissolved, with the PB buffer solution purification by liquid extraction product of pH 10.0, is collected
20 mL of water layer CH2Cl2Extraction.Finally organic phase is collected, and uses anhydrous Na2SO4Dry, rotary evaporation removes molten
Agent, vacuum drying obtain the product N of brown3-a-PEG21-a-N3(1.68 g, yield: 63.9%).
Example IV: the polymer poly (SS- of alkynyl sealing endalt-A)nSynthesis
Under nitrogen protection, by 3,3 '-two sulphur diine butyl dipropionate (B-ss- B) and two azidoethyls, two acetal radical
Polyethylene glycol (N3-a-PEG21-a-N3) chemically react, prepare the polymer poly (SS- of alkynyl sealing endalt-A)n.Specifically
Synthetic method is as follows:
Under the protection of nitrogen, addition CuBr(0.085 g in the side-neck flask of the DMF of 10 mL of Xiang Hanyou, 0.59
Mmol) and pentamethyl-diethylenetriamine (PMDETA) (0.102 g, 0.59 mmol), stirring both makes to be complexed, then according to
It is secondary that the B- synthesized is addedss- B(0.25 g, 0.79 mmol) and N3-a-PEG21-a-N3(0.97 g, 0.79 mmol).Pumping is filled
Gas three times after, device, which is sealed to be placed in 50 °C of oil baths under nitrogen protection, reacts 48 h, then, adds the B- of 10 mol%ss- B the reaction was continued 24 h, to ensure both ends as butynyl.Crude product is dialysed with DMF, and (dialysis bag retention molecular weight is
3500 Da), the dialyzate renewed at regular intervals.After 48 h, DMF is removed under reduced pressure in 60 °C of oil pumps.Then with 150 mL's
CH2Cl2It re-dissolves, saturation NaCl aqueous solution is added and removes mantoquita.Isolated organic layer anhydrous Na2SO4Dry 4 h
Afterwards, rotary evaporation removes solvent, then precipitates 3 times in diethyl ether solution, and sediment is collected and is dried in vacuo, viscosity is finally obtained
Product poly (SS-alt-A)n(0.74 g, yield: 60.7%).
Embodiment five: the adriamycin prodrug DOX- of double-responsehyd-poly(SS-alt-A)n-hydThe synthesis of-DOX
Utilize the doxorubicin derivative and polymer poly (SS- of desalting processingalt-A)nIt chemically reacts, is gone back
Original/pH dual responsiveness adriamycin prodrug DOX-hyd-poly(SS-alt-A)n-hyd-DOX.Specific synthetic method is as follows:
Firstly, to doxorubicin derivative (N3-hyd- DOX × HCl) carry out de- HCl treatment.In round-bottomed flask, it is added
N3-hyd- DOX × HCl(0.095 g, 0.13 mmol), it is dissolved with 3 mL DMSO, and the stirring of 10 mL triethylamines (TEA) is added
2 h siphon away supernatant after standing and add new TEA solution and continue to stir, are so repeated 2 times, finally leave and take DMSO layers of sample
Product are stand-by.In side-neck flask, lead in the state of nitrogen, CuBr(0.015 g, 0.104 mmol be first added) and PMDETA
(0.018 g, 0.104 mmol), is dissolved with 5 mL DMF solvents and stirs 5 min, then poly (SS- is successively added into bottlealt-A)n(0.23 g, 0.052 mmol) and the above-mentioned N for sloughing hydrochloride3-hydThe solution of-DOX will after taking out inflation three times
Bottle seal, which is placed in 50 °C of oil baths, reacts 48 h.After reaction, reaction solution is moved on to molecular cut off is 5000 Da's
It in bag filter, is dialysed using deionized water, a deionized water is changed after 4 h.It dialyses after 72 h, it will be molten in bag filter
Liquid takes out, and freezing, which is placed in freeze dryer, in refrigerator is lyophilized, and finally obtains Red viscous solid product DOX-hyd-poly
(SS-alt-A)n-hyd- DOX(0.26 g, yield: 84.2%).
Be utilized respectively nuclear magnetic resonance spectroscopy (1H NMR) and it is infrared spectroscopy (FT-IR) verifying gained small molecule compound, poly-
Close object and reduction/pH dual responsiveness adriamycin prodrug structure based on polyethylene glycol.Attached drawing 1 is doxorubicin derivative
(N3-hyd- DOX × HCl) hydrogen nuclear magnetic resonance spectrogram, attached drawing 2 be 3,3 '-two sulphur diine butyl dipropionate of polymer core
Magnetic resonance hydrogen spectrogram, attached drawing 3 are Cl-a-PEG21-a- Cl and N3-a-PEG21-a-N3Hydrogen nuclear magnetic resonance spectrogram, attached drawing 4 be poly-
Close object poly (SS-alt-A)3.6Hydrogen nuclear magnetic resonance spectrogram, attached drawing 5 be DOX-hyd-poly(SS-alt-A)3.6-hyd-DOX
Hydrogen nuclear magnetic resonance spectrogram, attached drawing 6 be HO-PEG21-OH, Cl-a-PEG21-a-Cl, N3-a-PEG21-a-N3, poly(SS-alt-A)3.6, DOX-hyd-N3And DOX-hyd-poly(SS-alt-A)3.6-hydThe infrared spectrogram of-DOX.In conjunction with attached drawing 1,
2,3,4,5,6 can meet experimental design with proved response product.
Embodiment six: polymeric prodrugs micella is prepared using dialysis
Take polymeric prodrugs (DOX-hyd-poly(SS-alt-A)3.6-hyd- DOX) 25 mg are dissolved in the DMF of 2 mL,
After stirring 2 h, with micro syringe with 2 mL h-1Speed the deionized water of 20 mL is added dropwise into solution, after completion of dropwise addition,
Continue to stir 2 h.Then, solution is fitted into the bag filter that molecular cut off is 5000 Da, with 24 h of deionized water dialysis
Afterwards, solution is taken out to and is settled to 25 mL, finally obtaining concentration is 1 mg mL-1Micellar solution.Attached drawing 7 is DOX-hyd-
poly(SS-alt-A)3.6-hydThe transmission electron microscope photo (A) and dynamic light scattering for the micella that-DOX is self-assembly of in water are bent
Line (B)；Such as: attached drawing 7(A) it show the pattern that polymeric prodrugs are self-assembly of spherical micelle in aqueous solution；Attached drawing 7(B)
It show the dynamic light scattering test curve figure of corresponding micellar particle size, it can be seen that polymeric prodrugs micella average grain diameter is 48
nm。
Embodiment seven: the vitro drug release test of polymeric prodrugs
(micellar concentration is 1 mg mL to the micellar solution for taking 4 mL well prepared in advance-1) molecular cut off is placed in as 7000 Da
Bag filter in, both ends, which are sealed, to be placed in the big centrifuge tube of 50 mL, then, 30 mL is separately added into corresponding centrifuge tube not
Same buffer solution.Buffer solution is divided into four kinds: (1) phosphate buffer (7.4,10 mM of pH)；(2) GSH's containing 10 mM
Phosphate buffer (7.4,10 mM of pH)；(3) acetate buffer solution (5.0,10 mM of pH)；(4) acetic acid of the GSH containing 10 mM
Buffer (5.0,10 mM of pH).Then, big centrifuge tube is placed in 37.5 °C of constant temperature oscillation instrument, with 160 r/min speed
It is vibrated.At the time point of setting, successively takes out the release liquid of 5 mL and add corresponding buffer solution.Every group of experiment carries out
3 balance tests, are finally averaged.The release liquid of taking-up is measured DOX concentration with sepectrophotofluorometer.Be loaded with Ah
Release profiles of the polymer micelle of mycin under the conditions of different reduction and pH are as shown in Fig. 8, in pH 5.0 or containing
In the phosphate buffer for having the GSH of 10 mM, drug release rate is significantly faster than that under normal physiological conditions；Meanwhile containing 10
It is maximum to add up release rate for drug in 5.0 buffer solution of pH of the GSH of mM.Drug release illustrates that the polymeric prodrugs micella has
Apparent reduction and pH dual responsiveness, can achieve the purpose of control release.
Embodiment eight: polymeric prodrugs micella inhibits tumor cell proliferation performance test
Human cervical carcinoma cell (HeLa cells) and human liver cancer cell (HepG2 cells) culture are being supplemented with respectively
In the DMEM culture medium of 10% fetal calf serum (FBS), 37 DEG C are placed in, 5% CO2It is cultivated in the incubator of (relative humidity 90%),
Regularly replace culture solution.Selection is in the cell inoculation of active growth phase and contains 96 holes of 100 μ L DMEM culture mediums in every hole
In plate, 24 h are cultivated.Polymeric prodrugs DOX- is configured with dialysishyd-poly(SS-alt-A)3.6-hydThe micella of-DOX is female
(doxorubicin concentration is 94 mg L to liquid-1), a series of micellar solution of various concentrations is added in 96 orifice plates, continues to train respectively
Support 48 h and 72 h.It is subsequently added into the MTT reagent of 25 μ L, after further cultivating 4 h, with microplate reader (Bio-Rad model
680) corresponding absorbance is measured at 570 nm.The calculation method of cell survival rate are as follows: cell survival rate (Cell
viability) (%) = [A]test/[A]control× 100%, wherein [A]testFor DOX-hyd-poly(SS-alt-
A)3.6-hydThe absorbance measured in the presence of-DOX polymeric prodrugs micella, and [A]controlThe case where for without polymeric prodrugs
Under the absorbance that measures.Each sample test three times, takes its average value.As shown in Fig. 9, (A) and (B) is respectively thin with HeLa
Born of the same parents cultivate the survival rate of cell after 48 h and 72 h；(C) and (D) is respectively and cell after HepG2 cell culture 48 h and 72 h
Survival rate.As the result is shown: compared with anticancer drugs, doxorubicin, synthesized polymeric prodrugs have good inhibition HeLa thin
The ability of born of the same parents and HepG2 cell Proliferation.
The method comprises the steps of firstly, preparing doxorubicin derivative (N3-hyd- DOX × HCl)；Prepare small point of alkynyl sealing end respectively again
Sub- 3,3 '-two sulphur diine butyl dipropionate (B- of compoundss- B) and azido sealing end two azidoethyl of compound, two acetal
Base polyethylene glycol (N3-a-PEG-a-N3)；Then, the polymer poly containing acetal bonds and disulfide bond is prepared by polymerization reaction
(SS-alt-A)；Finally, adriamycin is connected to polymer both ends by reaction again, obtaining has reduction and pH double-response
Adriamycin prodrug DOX-hyd-poly(SS-alt-A)3.6-hyd- DOX, other catalyst and ligand that the replacement present invention limits
Same available product, the material rate that the molecular weight and the present invention for replacing polyethylene glycol limit, available difference m value
The product of n value；What the present invention limited there is stimulating responsive polymer prodrug can be assembled into prodrug micelle simultaneously in aqueous solution
It is stabilized, and under acid either reducing condition, sensitive groups or segment can occur fracture and be destroyed micella, thus
The drug that can be will accumulate in inside micella rapidly releases, and for treating, achieves the technology effect recorded such as embodiment
Fruit.
Claims (10)
1. a kind of reduction/pH dual responsiveness adriamycin prodrug, which is characterized in that expressed by the following chemical structure formula:
In formula, m is that 3~113, n is 3~15.
2. reduction/pH dual responsiveness adriamycin prodrug according to claim 1, which is characterized in that the reduction/pH is dual
The preparation method of responsiveness adriamycin prodrug the following steps are included:
(1) using 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride as raw material, glacial acetic acid is catalyst, obtains Ah mould by reaction
Plain derivative；
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and ligand, with 3,3 '-two sulphur diine butyl dipropyls
Acid esters and two azidoethyls, two acetal radical polyethylene glycol are raw material, are replaced by the polyethylene glycol of reaction preparation two end alkynyl radicals sealing end
Copolymer；
(3) in the presence of copper salt catalyst and ligand, the polyethylene glycol of the two end alkynyl radicals sealing end obtained with step (2) replaces
Copolymer is reacted with the doxorubicin derivative that step (1) obtains, before the reduction/pH dual responsiveness adriamycin is prepared
Medicine.
3. reduction/pH dual responsiveness adriamycin prodrug according to claim 2, which is characterized in that in step (1), 6- bromine
Methyl caproate and reaction of sodium azide obtain 6- nitrine methyl caproate, and then 6- nitrine methyl caproate and hydration hydrazine reaction, obtain
6- nitrine hexanoyl hydrazine；It is raw material, 1- ethyl-(3- bis- using 3,3 '-dithiodipropionic acids and 3- butyne-1-ol in step (2)
Dimethylaminopropyl) carbodiimide hydrochloride is activator, 4-dimethylaminopyridine is catalyst, passes through esterification, it obtains
3,3 '-two sulphur diine butyl dipropionates；Polyethylene glycol and 2- chloroethyl vinyl ether is used to exist for raw material, acid catalyst
Under, reaction obtains the polyethylene glycol product that end is chloro, and then end is anti-for the polyethylene glycol product and sodium azide of chloro
It should obtain two azidoethyls, two acetal radical polyethylene glycol；In step (3), it will be used to react after doxorubicin derivative desalting processing.
4. reduction/pH dual responsiveness adriamycin prodrug according to claim 2, it is characterised in that: in step (1), reaction
Temperature be 60 DEG C~80 DEG C, time of reaction is the h of 12 h~24；In step (2), inert gas is nitrogen；The temperature of reaction
It is 25 DEG C~50 DEG C, the time of reaction is the h of 24 h~72；In step (3), the temperature of reaction is 25 DEG C~50 DEG C, reaction when
Between be the h of 24 h~48.
5. reduction/pH dual responsiveness adriamycin prodrug according to claim 2, it is characterised in that: copper salt catalyst is selected from
Salzburg vitriol, stannous chloride or cuprous bromide；Ligand be selected from sodium ascorbate, bipyridyl, five methyl diethylentriamine,
One of tetramethylethylenediamine or hexamethyl trien.
6. reduction/pH dual responsiveness adriamycin prodrug according to claim 2, it is characterised in that: in step (1), 6- is folded
The molar ratio of nitrogen hexanoyl hydrazine and doxorubicin hydrochloride is 1:(2~6)；In step (2), the poly- second two of two azidoethyls, two acetal radical
The molar ratio of alcohol, 3,3 '-two sulphur diine butyl dipropionates and copper salt catalyst is 1: (1.1~1.5): (0.5~1.5)；Copper
The molar ratio of salt catalyst and ligand is 1: (1~2)；In step (3), the polyethylene glycol alternating copolymerization of two end alkynyl radicals sealing end
Object, doxorubicin derivative molar ratio be 1:(2~5).
7. reduction/pH dual responsiveness adriamycin prodrug according to claim 3, it is characterised in that: every step after reaction,
All pass through purification processes.
8. a kind of preparation method of prodrug micelle, which comprises the following steps:
(1) using 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride as raw material, glacial acetic acid is catalyst, obtains Ah mould by reaction
Plain derivative；
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and ligand, with 3,3 '-two sulphur diine butyl dipropyls
Acid esters and two azidoethyls, two acetal radical polyethylene glycol are raw material, and the polyethylene glycol of two end alkynyl radicals sealing end is prepared by reacting
Alternate copolymer；
(3) in the presence of copper salt catalyst and ligand, the polyethylene glycol of the two end alkynyl radicals sealing end obtained with step (2) is handed over
It is reacted for copolymer with the doxorubicin derivative that step (1) obtains, reduction/pH dual responsiveness adriamycin prodrug is prepared；
(4) reduction/pH dual responsiveness adriamycin prodrug is self-assembly of prodrug micelle in aqueous solution；
The reduction/pH dual responsiveness adriamycin prodrug is expressed by the following chemical structure formula:
In formula, m is that 3~113, n is 3~15.
9. a kind of preparation method of stimulating responsive anticancer nano drug system, which comprises the following steps:
(1) using 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride as raw material, glacial acetic acid is catalyst, obtains Ah mould by reaction
Plain derivative；
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and ligand, with 3,3 '-two sulphur diine butyl dipropyls
Acid esters and two azidoethyls, two acetal radical polyethylene glycol are raw material, and the polyethylene glycol of two end alkynyl radicals sealing end is prepared by reacting
Alternate copolymer；
(3) in the presence of copper salt catalyst and ligand, the polyethylene glycol of the two end alkynyl radicals sealing end obtained with step (2) is handed over
It is reacted for copolymer with the doxorubicin derivative that step (1) obtains, reduction/pH dual responsiveness adriamycin prodrug is prepared；
(4) reduction/pH dual responsiveness adriamycin prodrug is self-assembly of prodrug micelle in aqueous solution；
(5) prodrug micelle and buffer are mixed to get stimulating responsive anticancer nano drug system；
The reduction/pH dual responsiveness adriamycin prodrug is expressed by the following chemical structure formula:
In formula, m is that 3~113, n is 3~15.
10. reduction described in claim 1/pH dual responsiveness adriamycin prodrug is in preparing stimulating responsive anticancer nano drug
Application.
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| CN110591078A (en) | 2019-12-20 |
| CN107141323A (en) | 2017-09-08 |
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