CN104027813A

CN104027813A - pH/reduction dual sensitive hydrophilic copolymer drug carrier as well as synthesis method and application thereof

Info

Publication number: CN104027813A
Application number: CN201410267878.1A
Authority: CN
Inventors: 徐明辉; 钱军民; 胥伟军; 柳雪峰; 王红洁
Original assignee: Xian Jiaotong University
Current assignee: Xian Jiaotong University
Priority date: 2014-06-16
Filing date: 2014-06-16
Publication date: 2014-09-10
Anticipated expiration: 2034-06-16
Also published as: CN104027813B

Abstract

The invention relates to a pH/reduction dual sensitive hydrophilic copolymer drug carrier as well as a synthesis method and application thereof. S,S'-bis(alpha, alpha'-dimethyl-alpha''-acetic acid) trithiocarbonate is taken as a chain transfer agent, RAFT (reversible additive fragment transfer) polymerization is carried out on N-t-butyloxycarboryl propylene hydrazine, N-(3-dimethylamine propyl) methacrylamide and N-hydroxymethyl acrylamide in two steps, reaction is carried out on hydroxyl on N-hydroxymethyl acrylamide and methoxyl polyethylene glycol at an isocyanic acid esterification end, polyethylene glycol is connected by virtue of a disulfide bond, and the target product, namely pH/reduction dual sensitive hydrophilic copolymer drug carrier, is obtained after a protecting group is removed. The pH/reduction dual sensitive hydrophilic copolymer drug carrier can load adriamycin and a gene drug at the same time; meanwhile, a hydrophilic polyethylene glycol shell can protect gene substance, long circulation is realized, a disulfide bond connected with polyethylene glycol is beneficial to escape of endosome of drug-carrier, the copolymer after being loaded with a drug can be self assembled into nanomicelle in water, bioavailability of the drug can be improved, and toxic and side effects can be reduced. The copolymer synthesis method provided by the invention has the advantages that molecular weight of each segment and quantity of polyethylene glycol segments can be flexibly adjusted and reaction conditions are mild.

Description

A kind of hydrophilic copolymers pharmaceutical carrier and synthetic method and application of pH/ reduction Dual Sensitive

Technical field

The invention belongs to biological medicament Material Field, be specifically related to a kind of hydrophilic copolymers pharmaceutical carrier and synthetic method and application of pH/ reduction Dual Sensitive.

Background technology

Malignant tumor has become the commonly encountered diseases frequently-occurring disease of serious harm human health, and the primary treatment means that adopt are at present surgical operation, three kinds of strategies of radiation and chemotherapy, and wherein chemotherapy is the inevitable choice when operation is invalid.Chemotherapy refers to utilizes cell toxicity medicament to treat the method for tumor.Yet chemotherapy of tumors curative effect has the problems such as strong dosage correlation, serious toxic and side effects and tumor multidrug-resistance, become the successful major obstacle of restriction chemotherapy of tumors.Oncotherapy is the challenging global problem of tool in contemporary medical science science.In recent years research shows, chemotherapeutics is loaded in the carrier of certain forms, can effectively improve oncotherapy effect, reduce toxic and side effects.Its reason is, the carrier of drug loading can the extension body internal recycle time, increase drug half-life, through tumor tissues special ' infiltration of enhancing and retention effect ', be enriched in tumor tissues, so both the ability of killing tumor cell can be improved, and the toxic and side effects of normal tissue and cell can be reduced again.Therefore, find the emphasis that desirable pharmaceutical carrier is tumor chemotherapeutic drug development always.

Amphipathic nature block polymer based on Polyethylene Glycol forms the hot issue that nano-micelle is current chemotherapeutics research.Because Polyethylene Glycol is a kind of hydrophilic polymer, there is biocompatibility excellence, non-immunogenic and the blood constituent advantage such as little that interacts, be that pharmaceutical carrier is modified indispensable important component part.Polyethyleneglycol modified nano-carrier can be avoided immune opsonic action, improves long circulation time in body, and can passive target in tumor tissues.Amphipathic nature block polymer normally be take Polyethylene Glycol as hydrophilic section, polyester as polylactide, polycaprolactone, poly (glycolide-lactide) and polyurethane, Merlon and hydrophobicity polyamino acid derivative be that hydrophobic section forms.When the concentration of these amphipathic copolymers in water is greater than critical micelle concentration, can be self-assembled into through hydrophobic interaction, electrostatic interaction, hydrogen bond and Van der Waals force etc. the nano-micelle of nucleocapsid structure, wherein hydrophobic section forms kernel, for hydrophobicity chemotherapeutic parcel, improve dissolubility in water, hydrophilic section forms shell, plays and stablizes micelle and long circulating action, improves drug bioavailability.The mode that polymer micelle loads hydrophobic drug mainly contains physically trapping and chemical bonding two classes.Wherein, physically trapping method utilizes similar compatibility principle between hydrophobic section and dewatering medicament to realize, and technical process is comparatively simple, but exists while sending in body the seepage may; And chemical bonding method drug loading can be avoided the leakage problems in delivery process, but its rate of release in target cell is often not fully up to expectations.For this reason, by utilizing, the chemical bond of tumor cell microenvironment sensitivity is embedded to carrier, become study hotspot.The Chinese patent that is 201210003860.1 as application number discloses a kind of nano-micelle prodrug that utilizes disulfide bond to connect Polyethylene Glycol section and year camptothecine Polyaspartimides section.Although this carrier can dissociate in tumor cell, medicine still, on polymer, is difficult to give full play to curative effect.

Gene therapy, as a kind of new technique means of capturing malignant tumor, is sent to great expectations.Gene therapy is by therapeutic genes is imported in target cell, with correcting defect gene, inhibition or strengthen certain genes/proteins matter and express, reaches a class technology for the treatment of disease object, mainly comprises DNA, siRNA and miRNA etc.Yet exposed genetic stew bear electricity, be difficult to cross over after birth structure, and Half-life in vivo is very short, very easily cannot be arrived diseased region by regulation of immune system, seriously restricts the clinical practice of gene therapy.Therefore, develop the key that suitable delivery vector has become gene therapy.With respect to traditional virus gene carrier, the non-viral vector based on cation high molecular has advantage at aspects such as safety, immunogenicity, preparation complexities, is expected to replace viral vector.Yet cationic polymer carrier exists the positively related transfection efficiency of molecular weight and cytotoxicity, therefore extremely important to the control of its molecular weight.

Summary of the invention

The object of the present invention is to provide a kind of hydrophilic copolymers pharmaceutical carrier and synthetic method and application that can improve curative effect of medication and reduce the pH/ reduction Dual Sensitive of toxic and side effects, the hydrophilic copolymers pharmaceutical carrier of the pH/ reduction Dual Sensitive that the method is synthetic can load amycin and genetic stew simultaneously, and after loading amycin, from hydrophilic, become amphipathicly, can self assembly form nanoparticle.

In order to achieve the above object, the synthetic method of the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive of the present invention, comprises the steps:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

Tertbutyloxycarbonyl hydrazine is dissolved in the water of 0 ℃～4 ℃, then under agitation drip the organic solution of acryloyl chloride, regulate pH value to 8～9 simultaneously, again room temperature reaction 4～10 hours, finally by the product obtaining through dichloromethane extraction, dry, recrystallization process, obtain N-tertbutyloxycarbonyl propylene hydrazides; Wherein, the mol ratio of the acryloyl chloride in tertbutyloxycarbonyl hydrazine and acryloyl chloride organic solution is 1:1.1;

2) poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is synthetic:

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and initiator be dissolved in anhydrous and oxygen-free organic solvent; in 60～65 ℃ of reactions 24～72 hours, obtain reaction system, reaction system is concentrated; then to add dichloromethane to dissolve; add wherein cold diethyl ether to make to separate out precipitation, collecting precipitation, is gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents again; Wherein, S, the mol ratio of S'-two (α, α '-dimethyl-α " acetic acid) trithiocarbonate and N-tertbutyloxycarbonyl propylene hydrazides is 5:(40～300);

3) poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is synthetic:

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free organic solvent, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and initiator, in 60～65 ℃ of reactions 24～72 hours, obtain reaction system, reaction system is concentrated, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, is gathered (N-tertbutyloxycarbonyl propylene hydrazides)-is gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer; Wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:(0.3～0.8): (0.01～0.05);

4) poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is synthetic:

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is dissolved in to anhydrous organic solvent, then add wherein 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, in 60 ℃, react 36～72 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis, lyophilizing again, obtain poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, wherein, in poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that the end methoxy poly (ethylene glycol) that 2,2'-dithio diethyl is isocyanate-modified and process dewater-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, the mol ratio of pendant hydroxyl group is (2～5): 1,

5) the tertbutyloxycarbonyl protecting group of gathering in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is removed, obtain the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive.

Described step 1) solvent in the organic solution of acryloyl chloride is chloroform, dichloromethane or dioxane, step 2) and step 3) in anhydrous and oxygen-free organic solvent be anhydrous and oxygen-free dioxane, anhydrous and oxygen-free chloroform or anhydrous and oxygen-free oxolane; Step 4) anhydrous organic solvent in is anhydrous dimethyl formamide, anhydrous dioxane, anhydrous chloroform, dry toluene or anhydrous tetrahydro furan.

Described step 2) and step 3) in initiator be azodiisobutyronitrile or 4,4'-azo two (cyanopentanoic acid).

Described step 2) S in, two (the α of S'-, α '-dimethyl-α " acetic acid) mol ratio of trithiocarbonate and initiator is 5:1, step 3) and in the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents and initiator be 10:1.

Poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that described process dewaters-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is adopted dehydration with the following method and is obtained: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under nitrogen atmosphere, reflux and dewater 2～6 hours.

Described 2, the isocyanate-modified end methoxy poly (ethylene glycol) of 2'-dithio diethyl is adopted with the following method and is obtained: the end methoxy poly (ethylene glycol) that is 800～4000Da by molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with anhydrous n-hexane, be settled out solid, collect solid drying, obtain the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:(0.02～0.08): (3～8).

Described step 5) adopt and remove with the following method tertbutyloxycarbonyl protecting group: will gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer and be dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 4～10 hours, reaction finishes the rear concentrated reaction system obtaining, then with sodium bicarbonate solution, neutralize, then in the ammonia that utilizes dialyzer that molecular cut off is 3500Da to be 0.25% in mass concentration, dialyse, last lyophilizing, obtain the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive, wherein, the trifluoroacetic acid in mixed solution is (15～30) with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer: 1.

The hydrophilic copolymers pharmaceutical carrier of the pH/ reduction Dual Sensitive that the synthetic method described in a kind of employing makes, its chemical name is polyacrylic hydrazide-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, and chemical structural formula is:

The application of a kind of hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive in loading amycin and genomic medicine.

Described genomic medicine is siRNA.

Described step 2) in, in every 100 milliliters of anhydrous and oxygen-free organic solvents, dissolve the N-tertbutyloxycarbonyl propylene hydrazides of 10～40g.

Described step 3) in, in every 100 milliliters of anhydrous and oxygen-free organic solvents, dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents of 6～20g.

Described step 4) in, in every 100 milliliters of anhydrous organic solvents, dissolve 5～15g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer through dewatering.

In every 100 milliliters of dry toluenes, dissolve the end methoxy poly (ethylene glycol) of 5～30g.

Described step 5) in, in every 100 milliliters of mixed solvents, dissolve 5～25g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer.

Described backflow temperature for removing water is 110 ℃～120 ℃.

Compared with prior art, beneficial effect of the present invention is:

(1) the hydrophilic copolymers pharmaceutical carrier molecular structure of the synthetic pH/ of the present invention reduction Dual Sensitive is block copolymer and/or comb-shaped copolymer, by three kinds of hydrophilic component of polymer, formed, be respectively the Polyethylene Glycol that can realize recessive role in body, can load the polyacrylic hydrazide of amycin and can gather (N-(3-dimethylamine propyl) Methacrylamide) through the cation of electrostatic interaction load genetic stew by chemical bonding, therefore, the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive prepared by the present invention loads when can realize amycin and genomic medicine, realize synergistic antitumor treatment.

(2) the hydrophilic copolymers pharmaceutical carrier of this pH/ reduction Dual Sensitive is water miscible originally, by chemical bonding, after polyacrylic hydrazide section is loaded amycin, present amphipathic, it can self assembly form nano-micelle in aqueous solution, have through tumor tissues ' infiltration of enhancing and retention effect ' passive target function, be enriched in tumor tissues for oral administration, promote tumor cell picked-up, improve bioavailability.

(3) cation of the present invention poly-(N-(3-dimethylamine propyl) Methacrylamide) section can form complex by electrostatic interaction and bear electricity genetic stew, the compression of realization to genetic stew, be covered in hydrophobic carrying outside amycin kernel, compare and there is higher stability with simple cationic polymer carrier, effectively protect genetic stew; The quantity scalable of outermost layer Polyethylene Glycol, can more effectively regulate stability and body-internal-circulation time in blood; Such structural system will have lower critical micelle concentration, and in the time of can avoiding entering blood circulation, dilution effect causes the risk of micelle unstability.

(4) Polyethylene Glycol of the present invention is connected on copolymer by disulfide bond, in carrier enters tumor cell, can respond the acid environment of its reproducibility and dissociate and slough, the cationic layer exposing can promote carrier to escape and enter Cytoplasm from endosome, in addition, because amycin is by being bonded on carrier the hydrazone key of cell inner acidic environment sensitive on carrier, the hydrophilic copolymers pharmaceutical carrier that is loaded with the pH/ reduction Dual Sensitive of amycin can respond tumor cell sour environment, make amycin discharge fast at short notice the concentration that improves medicine, therefore, the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive prepared by the present invention can improve the drug effect of amycin, and reduction toxic and side effects.

(5) polyacrylic hydrazide and poly-(N-(3-dimethylamine propyl) Methacrylamide) are nondegradable medicine carrying section polymer; by RAFT polymerization, synthesized; its molecular size range can accurately be controlled and narrow molecular weight distribution; both can guarantee efficiency gene transfection; can guarantee again to excrete by Excretory system, avoid cylinder accumulation or cause negative consequence.

(6) advantages such as reaction condition of the present invention is gentle, raw material is easy to get, medicine stowage is simple, and applicable genetic stew kind is many.

Accompanying drawing explanation

Fig. 1 is the hydrophilic copolymers of pH/ reduction Dual Sensitive of the present invention and the route schematic diagram that medicine loads.

Fig. 2 is poly-(N-(3-dimethylamine propyl) Methacrylamide)-polyacrylic hydrazide-ethylene glycol copolymer of hydrophilic of the synthetic pH/ reduction Dual Sensitive of embodiment 1 ¹h-NMR spectrogram;

Fig. 3 is the particle size distribution figure that the hydrophilic copolymers pharmaceutical carrier of the synthetic pH/ of embodiment 1 reduction Dual Sensitive loads self-assembled nanometer grain after amycin;

Fig. 4 is that the hydrophilic copolymers pharmaceutical carrier loading amycin of the synthetic pH/ reduction Dual Sensitive of embodiment 1 and the nanoparticle of FAM labelling siRNA enter the laser co-focusing fluorescence photo in breast cancer cell MCF-7; Wherein, a is the nuclear fluorescence photo of DAPI dyeing, and b is the amycin fluorescence photo at nucleus place, and c is that the siRNA of FAM labelling enters the fluorescence photo after cell, the constitutional diagram that d is a-c.

The specific embodiment

Below by embodiment, the invention will be further described, but the present invention is not limited to this.

Embodiment 1:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

Tertbutyloxycarbonyl hydrazine is dissolved in to the solution that forms 1mol/L in the water of 0 ℃, then under vigorous stirring, in 30 minutes, drip the chloroformic solution that concentration is the acryloyl chloride of 5mol/L, utilize the sodium hydrate aqueous solution of 2mol/L to regulate pH value to 8～9 simultaneously, again room temperature reaction 10 hours, finally by the product obtaining through dichloromethane extraction, dry and recrystallization, obtain N-tertbutyloxycarbonyl propylene hydrazides; Wherein, the mol ratio of the acryloyl chloride in the chloroformic solution of tertbutyloxycarbonyl hydrazine and acryloyl chloride is 1:1.1; And recrystallization adopts dichloromethane recrystallization;

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile be dissolved in anhydrous and oxygen-free dioxane; and in 60 ℃ of reactions 48 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate; then add dichloromethane to dissolve; add wherein cold diethyl ether to make to separate out precipitation, collecting precipitation, is gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents again; Wherein, S, the mol ratio of S'-two (α, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile is 5:100:1; Every 100 milliliters of anhydrous and oxygen-free dioxane dissolve the N-tertbutyloxycarbonyl propylene hydrazides of 15g;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free dioxane, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and azodiisobutyronitrile, in 60 ℃ of reactions 48 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.8:0.05, and azodiisobutyronitrile is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, in every 100 milliliters of anhydrous and oxygen-free dioxane, dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents of 16g,

4) 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified synthetic:

The end methoxy poly (ethylene glycol) that is 2000Da by dry molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with anhydrous n-hexane, precipitate 3 times, after the solid drying obtaining, be the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:0.02:3, dissolves the end methoxy poly (ethylene glycol) of 30g in every 100 milliliters of dry toluenes;

5) poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is synthetic:

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is dissolved in anhydrous chloroform, then add wherein 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 60 ℃ are reacted 72 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtain poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, wherein, in poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that the end methoxy poly (ethylene glycol) that 2,2'-dithio diethyl is isocyanate-modified and process dewater-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, the mol ratio of pendant hydroxyl group is 2:1, in every 100 milliliters of anhydrous chloroforms, dissolve 10g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer through dewatering, through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, adopt dehydration with the following method and obtain: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, to reflux dewater obtains for 2 hours, and reflux temperature is 120 ℃,

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 10 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then in the ammonia that utilizes dialyzer that molecular cut off is 3500Da to be 0.25% in mass concentration, dialyse two days, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, trifluoroacetic acid in mixed solvent is 15:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, the volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, dissolves poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 25g in every 100 milliliters of mixed solvents.

The chemical name of the hydrophilic copolymers pharmaceutical carrier of the pH/ reduction Dual Sensitive that the present embodiment is synthetic is poly-(N-(3-dimethylamine propyl) Methacrylamide)-polyacrylic hydrazide-ethylene glycol copolymer ¹h-NMR spectrogram is shown in Fig. 2.As can be seen from Figure 2, the chemical shift of each copolymer component proton has all obtained good evaluation, shows that the synthetic of this copolymer is successful.The structural formula of this copolymer is:

Utilize the hydrophilic copolymers pharmaceutical carrier of the synthetic pH/ reduction Dual Sensitive of the present embodiment to load after amycin, present amphipathic character, can be self-assembled into as nanoparticle in water, the particle size distribution figure that dynamic light scattering method records is shown in Fig. 3.As seen from Figure 3, centralized particle diameter is in 100-200 nanometer, and particle size distribution range is narrow, more satisfactory.

The hydrophilic copolymers pharmaceutical carrier of the pH/ reduction Dual Sensitive that the present embodiment is synthetic can load amycin and siRNA simultaneously, its stowage is that the hydrophilic copolymers pharmaceutical carrier of the pH/ reduction Dual Sensitive after genomic medicine and loading amycin is mixed, through vortex agitator, be mixed to form complex, realize the loading of genomic medicine.

Take and load amycin and bcl-2siRNA is example simultaneously, the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive is carried to the laser co-focusing fluorescence photo that the nanoparticle that forms after the bcl-2siRNA of amycin and green fluorescence agent FAM labelling enters in breast cancer cell MCF-7 altogether and analyze, Fig. 4 is shown in the distribution in breast cancer cell MCF-7.B in Fig. 4 a is the nuclear fluorescence sending of DAPI dyeing, R in Fig. 4 b is the fluorescence that the amycin at nucleus place sends, G in Fig. 4 c is that the siRNA of FAM labelling enters the fluorescence sending after cell, to after Fig. 4 a-4c combination, obtain Fig. 4 d, by Fig. 4 d, can be found out, the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive carries altogether the nanoparticle forming after the bcl-2siRNA of amycin and green fluorescence agent FAM labelling and can enter in breast cancer cell MCF-7 through thin endocytosis, and amycin and siRNA all can effectively enter in breast cancer cell MCF-7, be distributed in around the nucleus of breast cancer cell MCF-7, visible synthetic carrier can be used for the common loading of amycin and siRNA and sends.

Embodiment 2:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

Tertbutyloxycarbonyl hydrazine is dissolved in to the solution that forms 1mol/L in the water of 2 ℃, then under vigorous stirring, in 30 minutes, drip the dichloromethane solution that concentration is the acryloyl chloride of 5mol/L, utilize the sodium hydrate aqueous solution of 2mol/L to regulate pH value to 8～9 simultaneously, again room temperature reaction 8 hours, finally will obtain product through dichloromethane extraction, dry and recrystallization process, obtain N-tertbutyloxycarbonyl propylene hydrazides; Wherein, the mol ratio of the acryloyl chloride in the dichloromethane solution of tertbutyloxycarbonyl hydrazine and acryloyl chloride is 1:1.1; And recrystallization adopts dichloromethane recrystallization;

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile be dissolved in anhydrous and oxygen-free dioxane; and in 60 ℃ of reactions 24 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate; then add dichloromethane to dissolve; add wherein cold diethyl ether to make to separate out precipitation, collecting precipitation, is gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents again; Wherein, S, the mol ratio 5:300:1 of S'-two (α, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile; Every 100 milliliters of anhydrous and oxygen-free dioxane dissolve 20g N-tertbutyloxycarbonyl propylene hydrazides;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free dioxane, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and azodiisobutyronitrile, in 60 ℃ of reactions 72 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.3:0.01, and azodiisobutyronitrile is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, in every 100 milliliters of anhydrous and oxygen-free dioxane, dissolve N-(3-dimethylamine propyl) Methacrylamide of 20g,

The end methoxy poly (ethylene glycol) that is 800Da by dry molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with precipitating in anhydrous n-hexane 3 times, after the solid drying obtaining, be the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:0.08:8; In every 100 milliliters of dry toluenes, dissolve the end methoxy poly (ethylene glycol) of 5g;

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is dissolved in to anhydrous dimethyl formamide, then add wherein 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 60 ℃ are reacted 36 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtain poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, wherein, 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified and be 5:1 through the mol ratio of pendant hydroxyl group in poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, dissolves 15g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer through dewatering in every 100 milliliters of anhydrous dimethyl formamides, through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, adopt dehydration with the following method and obtain: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, refluxing dewaters obtains for 2 hours, reflux temperature is that reflux temperature is 120 ℃,

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 6 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then in the ammonia that utilizes dialyzer that molecular cut off is 3500Da to be 0.25% in mass concentration, dialyse two days, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, trifluoroacetic acid in mixed solvent is 30:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, the volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, every 100 milliliters of mixed solvents dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 15g.

Embodiment 3:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

Tertbutyloxycarbonyl hydrazine is dissolved in to the solution that forms 1mol/L in the water of 4 ℃, then under vigorous stirring, in 30 minutes, drip the dioxane solution that concentration is the acryloyl chloride of 5mol/L, utilize the sodium hydrate aqueous solution of 2mol/L to regulate pH value to 8～9 simultaneously, again room temperature reaction 4 hours, finally by the product obtaining through dichloromethane extraction, dry and recrystallization process, obtain N-tertbutyloxycarbonyl propylene hydrazides; Wherein, the mol ratio of the acryloyl chloride in the dioxane solution of tertbutyloxycarbonyl hydrazine and acryloyl chloride is 1:1.1; And recrystallization adopts dichloromethane recrystallization;

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4,4'-azo two (cyanopentanoic acid) be dissolved in anhydrous and oxygen-free dioxane, in 60 ℃ of reactions 60 hours; obtain reaction system; reaction system distilling under reduced pressure is made to concentrate, then adds dichloromethane to dissolve, then add wherein cold diethyl ether to make to separate out precipitation; collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents; Wherein, S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4; mol ratio of 4'-azo two (cyanopentanoic acid) is 5:200:1, every 100 milliliters of anhydrous and oxygen-free dioxane dissolve 40g N-tertbutyloxycarbonyl propylene hydrazides;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free dioxane, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and 4, 4'-azo two (cyanopentanoic acid), in 60 ℃ of reactions 72 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.4:0.02,4,4'-azo two (cyanopentanoic acid) is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, dissolves poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents of 8g in every 100 milliliters of anhydrous and oxygen-free dioxane,

The end methoxy poly (ethylene glycol) that is 800Da by dry molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with precipitating in anhydrous n-hexane 3 times, after the solid drying obtaining, obtain the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:0.06:7, dissolves the end methoxy poly (ethylene glycol) of 12g in every 100 milliliters of dry toluenes;

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer is dissolved in dry toluene, then add wherein 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 60 ℃ are reacted 36 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtain poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, wherein, 2, in poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified and process dewater-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, the mol ratio of pendant hydroxyl group is 4:1, in every 100 milliliters of dry toluenes, dissolve 5g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer through dewatering, through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, adopt dehydration with the following method and obtain: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, to reflux dewater obtains for 4 hours, and reflux temperature is 120 ℃,

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 8 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then in the ammonia that utilizes dialyzer that molecular cut off is 3500Da to be 0.25% in mass concentration, dialyse two days, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, trifluoroacetic acid in mixed solvent is 18:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, the volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, every 100 milliliters of mixed solvents dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 20g.

Embodiment 4:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

Tertbutyloxycarbonyl hydrazine is dissolved in to the solution that forms 1mol/L in the water of 0 ℃, then under vigorous stirring, in 30 minutes, drip the dichloromethane solution that concentration is the acryloyl chloride of 5mol/L, utilize 2mol/L sodium hydrate aqueous solution to regulate pH value to 8～9 simultaneously, again room temperature reaction 9 hours, finally by the product obtaining through dichloromethane extraction, dry and recrystallization process, obtain N-tertbutyloxycarbonyl propylene hydrazides; Wherein, the mol ratio of the acryloyl chloride in the dichloromethane solution of tertbutyloxycarbonyl hydrazine and acryloyl chloride is 1:1.1; And recrystallization adopts dichloromethane recrystallization;

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4,4'-azo two (cyanopentanoic acid) be dissolved in anhydrous and oxygen-free oxolane, in 60 ℃ of reactions 55 hours; obtain reaction system; reaction system distilling under reduced pressure is made to concentrate, then adds dichloromethane to dissolve, then add wherein cold diethyl ether to make to separate out precipitation; collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents; Wherein, S, S'-two (α, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4; mol ratio of 4'-azo two (cyanopentanoic acid) is 5:150:1, and every 100 milliliters of anhydrous and oxygen-free oxolanes dissolve the N-tertbutyloxycarbonyl propylene hydrazides of 35g;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free oxolane, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and 4, 4'-azo two (cyanopentanoic acid), in 60 ℃ of reactions 72 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.7:0.03,4,4'-azo two (cyanopentanoic acid) is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, in every 100 milliliters of anhydrous and oxygen-free oxolanes, dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents of 9g,

The end methoxy poly (ethylene glycol) that is 4000Da by dry molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with precipitating in anhydrous n-hexane 3 times, after the solid drying obtaining, obtain the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:0.05:5, dissolves the end methoxy poly (ethylene glycol) of 5g in every 100 milliliters of dry toluenes;

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer is dissolved in anhydrous chloroform, then add 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 60 ℃ are reacted 50 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtains poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer; Wherein, 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified and be 5:1 through the mol ratio of pendant hydroxyl group in poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, dissolves 14g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer through dewatering in every 100 milliliters of anhydrous chloroforms; And adopt dehydration with the following method and obtain through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, refluxing dewaters obtains for 5 hours; And reflux temperature is 115 ℃;

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 6 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then in the ammonia that utilizes dialyzer that molecular cut off is 3500Da to be 0.25% in mass concentration, dialyse two days, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, the trifluoroacetic acid in mixed solvent is 25:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer; The volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, every 100 milliliters of mixed solvents dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 7g.

Embodiment 5:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

Tertbutyloxycarbonyl hydrazine is dissolved in to the solution that forms 1mol/L in the water of 4 ℃, then under vigorous stirring, in 30 minutes, drip the dioxane solution that concentration is the acryloyl chloride of 5mol/L, utilize the sodium hydrate aqueous solution of 2mol/L to regulate pH value to 8～9 simultaneously, again room temperature reaction 4.5 hours, finally by the product obtaining through dichloromethane extraction, dry and recrystallization process, obtain N-tertbutyloxycarbonyl propylene hydrazides; Wherein, the mol ratio of the acryloyl chloride in the dioxane solution of tertbutyloxycarbonyl hydrazine and acryloyl chloride is 1:1.1; And recrystallization adopts dichloromethane recrystallization;

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile be dissolved in anhydrous and oxygen-free chloroform; and in 60 ℃ of reactions 50 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate; then add dichloromethane to dissolve; add wherein cold diethyl ether to make to separate out precipitation, collecting precipitation, is gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents again; Wherein, S, the mol ratio of S'-two (α, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile is 5:260:1, every 100 milliliters of anhydrous and oxygen-free chloroforms dissolve 35g N-tertbutyloxycarbonyl propylene hydrazides;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free chloroform, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and azodiisobutyronitrile, in 60 ℃ of reactions 52 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.4:0.02, azodiisobutyronitrile is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, dissolves poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents of 9g in every 100 milliliters of anhydrous and oxygen-free chloroforms,

The end methoxy poly (ethylene glycol) that is 2000Da by dry molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with precipitating in anhydrous n-hexane 3 times, after the solid drying obtaining, obtain the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:0.06:8; In every 100 milliliters of dry toluenes, dissolve the end methoxy poly (ethylene glycol) of 5g;

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer is dissolved in anhydrous tetrahydro furan, then add wherein 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 60 ℃ are reacted 72 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtain poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, wherein, the mol ratio of the middle pendant hydroxyl group of poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that the end methoxy poly (ethylene glycol) that 2,2'-dithio diethyl is isocyanate-modified and process dewater-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is 3:1, every 100 milliliters of anhydrous tetrahydro furans dissolve 15g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer through dewatering, and through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, adopt dehydration with the following method and obtain: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, to reflux dewater obtains for 6 hours, and reflux temperature is 110 ℃,

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 8 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then utilizing the dialyzer that molecular cut off is 3500Da is in 0.25% ammonia, to dialyse two days in mass concentration, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, trifluoroacetic acid in mixed solvent is 21:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, the volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, every 100 milliliters of mixed solvents dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 20g.

Embodiment 6:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

Tertbutyloxycarbonyl hydrazine is dissolved in to the solution that forms 1mol/L in the water of 1 ℃, then under vigorous stirring, in 30 minutes, drip the chloroformic solution that concentration is the acryloyl chloride of 5mol/L, utilize 2mol/L sodium hydrate aqueous solution to regulate pH value to 8～9 simultaneously, again room temperature reaction 6 hours, finally by the product obtaining through dichloromethane extraction, dry and recrystallization process, obtain N-tertbutyloxycarbonyl propylene hydrazides; Wherein, the mol ratio of the acryloyl chloride in the chloroformic solution of tertbutyloxycarbonyl hydrazine and acryloyl chloride is 1:1.1; And recrystallization adopts dichloromethane recrystallization;

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4,4'-azo two (cyanopentanoic acid) be dissolved in anhydrous and oxygen-free dioxane, in 60 ℃ of reactions 55 hours; obtain reaction system; reaction system distilling under reduced pressure is made to concentrate, then adds dichloromethane to dissolve, then add wherein cold diethyl ether to make to separate out precipitation; collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents; Wherein, S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4; mol ratio of 4'-azo two (cyanopentanoic acid) is 5:120:1, the N-tertbutyloxycarbonyl propylene hydrazides of the 30g that every 100 milliliters of anhydrous and oxygen-free dioxane dissolve;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free dioxane, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and 4, 4'-azo two (cyanopentanoic acid), in 60 ℃ of reactions 24 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.7:0.04,4,4'-azo two (cyanopentanoic acid) is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, dissolves N-(3-dimethylamine propyl) Methacrylamide of 13g in every 100 milliliters of anhydrous and oxygen-free dioxane,

The end methoxy poly (ethylene glycol) that is 1200Da by dry molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with precipitating in anhydrous n-hexane 3 times, after the solid drying obtaining, be the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:0.04:6; In every 100 milliliters of dry toluenes, dissolve the end methoxy poly (ethylene glycol) of 30g;

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is dissolved in anhydrous dioxane, then add wherein 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 60 ℃ are reacted 68 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtain poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, wherein, the mol ratio of the pendant hydroxyl group in poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that the end methoxy poly (ethylene glycol) that 2,2'-dithio diethyl is isocyanate-modified and process dewater-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is 3.6:1, in every 100 milliliters of anhydrous dioxane, dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer that the process of 13g dewaters, through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, adopt dehydration with the following method and obtain: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, refluxing dewaters obtains for 4 hours, reflux temperature is that reflux temperature is 115 ℃,

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 5 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then utilizing the dialyzer that molecular cut off is 3500Da is in 0.25% ammonia, to dialyse two days in mass concentration, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, the trifluoroacetic acid in mixed solvent is 23:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer; The volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, every 100 milliliters of mixed solvents dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 11g.

Embodiment 7:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4,4'-azo two (cyanopentanoic acid) be dissolved in anhydrous and oxygen-free dioxane, in 65 ℃ of reactions 72 hours; obtain reaction system; reaction system distilling under reduced pressure is made to concentrate, then adds dichloromethane to dissolve, then add wherein cold diethyl ether to make to separate out precipitation; collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents; Wherein, S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4; mol ratio of 4'-azo two (cyanopentanoic acid) is 5:40:1, the N-tertbutyloxycarbonyl propylene hydrazides of the 22g that every 100 milliliters of anhydrous and oxygen-free dioxane dissolve;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free dioxane, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and 4, 4'-azo two (cyanopentanoic acid), in 65 ℃ of reactions 24 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.7:0.04,4,4'-azo two (cyanopentanoic acid) is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, dissolves N-(3-dimethylamine propyl) Methacrylamide of 6g in every 100 milliliters of anhydrous and oxygen-free dioxane,

The end methoxy poly (ethylene glycol) that is 2500Da by dry molecular weight, dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with precipitating in anhydrous n-hexane 3 times, after the solid drying obtaining, be the isocyanate-modified end methoxy poly (ethylene glycol) of 2,2'-dithio diethyl; Wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:0.04:6; In every 100 milliliters of dry toluenes, dissolve the end methoxy poly (ethylene glycol) of 30g;

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 4 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then utilizing the dialyzer that molecular cut off is 3500Da is in 0.25% ammonia, to dialyse two days in mass concentration, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, the trifluoroacetic acid in mixed solvent is 23:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer; The volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, every 100 milliliters of mixed solvents dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 5g.

Embodiment 8:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile be dissolved in anhydrous and oxygen-free chloroform; and in 60 ℃ of reactions 50 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate; then add dichloromethane to dissolve; add wherein cold diethyl ether to make to separate out precipitation, collecting precipitation, is gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents again; Wherein, S, the mol ratio of S'-two (α, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and azodiisobutyronitrile is 5:260:1, and every 100 milliliters of anhydrous and oxygen-free chloroforms dissolve the N-tertbutyloxycarbonyl propylene hydrazides of 30g;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free chloroform, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and azodiisobutyronitrile, in 63 ℃ of reactions 52 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.4:0.02, azodiisobutyronitrile is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, dissolves poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents of 9g in every 100 milliliters of anhydrous and oxygen-free chloroforms,

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer is dissolved in anhydrous tetrahydro furan, then add wherein 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 63 ℃ are reacted 72 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtain poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, wherein, the mol ratio of the middle pendant hydroxyl group of poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that the end methoxy poly (ethylene glycol) that 2,2'-dithio diethyl is isocyanate-modified and process dewater-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is 3:1, every 100 milliliters of anhydrous tetrahydro furans dissolve 15g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer through dewatering, and through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, adopt dehydration with the following method and obtain: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, to reflux dewater obtains for 6 hours, and reflux temperature is 110 ℃,

6) remove blocking group tertbutyloxycarbonyl in copolymer:

To gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer is dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 8 hours, reaction finishes the rear concentrated reaction system obtaining, then the sodium bicarbonate solution neutralization that is 1% by mass concentration, then utilizing dialyzer that molecular cut off is 3500Da is that 0.25% ammonia is analysed two days in molten in mass concentration, last lyophilizing, obtains the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive; Wherein, trifluoroacetic acid in mixed solvent is 21:1 with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, the volume ratio 1:1 of trifluoroacetic acid and dichloromethane in mixed solvent, every 100 milliliters of mixed solvents dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer of 20g.

Embodiment 9:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

By S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4,4'-azo two (cyanopentanoic acid) be dissolved in anhydrous and oxygen-free oxolane, in 60 ℃ of reactions 55 hours; obtain reaction system; reaction system distilling under reduced pressure is made to concentrate, then adds dichloromethane to dissolve, then add wherein cold diethyl ether to make to separate out precipitation; collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents; Wherein, S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate, N-tertbutyloxycarbonyl propylene hydrazides and 4; mol ratio of 4'-azo two (cyanopentanoic acid) is 5:150:1, every 100 milliliters of anhydrous and oxygen-free oxolanes dissolve 10g N-tertbutyloxycarbonyl propylene hydrazides;

To gather (N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents is dissolved in anhydrous and oxygen-free oxolane, then under nitrogen atmosphere, add N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide and 4, 4'-azo two (cyanopentanoic acid), in 65 ℃ of reactions 72 hours, obtain reaction system, reaction system distilling under reduced pressure is made to concentrate, then add dichloromethane to dissolve, add wherein again cold diethyl ether to make to separate out precipitation, collecting precipitation, gathered (N-tertbutyloxycarbonyl propylene hydrazides)-gathered (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, wherein, the mol ratio of the N-tertbutyloxycarbonyl propylene hydrazides construction unit in poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, N-(3-dimethylamine propyl) Methacrylamide, N hydroxymethyl acrylamide is 1:0.7:0.03,4,4'-azo two (cyanopentanoic acid) is 1:10 with the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents, in every 100 milliliters of anhydrous and oxygen-free oxolanes, dissolve poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents of 9g,

Poly-(N-tertbutyloxycarbonyl propylene hydrazides) through dewatering-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer is dissolved in anhydrous chloroform, then add 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, under nitrogen atmosphere, 65 ℃ are reacted 50 hours, in the reaction system obtaining, add cold diethyl ether to make precipitation, then to distill water dialysis 48 hours, lyophilizing again, obtains poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer; Wherein, 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified and be 5:1 through the mol ratio of pendant hydroxyl group in poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering, dissolves 14g poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) allylamine/N hydroxymethyl acrylamide) copolymer through dewatering in every 100 milliliters of anhydrous chloroforms; And adopt dehydration with the following method and obtain through poly-(N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer dewatering: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under blanket of nitrogen, refluxing dewaters obtains for 5 hours; And reflux temperature is 115 ℃;

6) remove blocking group tertbutyloxycarbonyl in copolymer:

Cold diethyl ether temperature in above-described embodiment 1-9 is subzero-20 ℃, and vacuum drying temperature is at 30 ℃, and freeze temperature is-40 ℃, when reaction system concentrates, is concentrated to 1/3rd to 1/2nd of original volume.In the present invention 2, the isocyanate-modified end methoxy poly (ethylene glycol) of 2'-dithio diethyl is that application number is 201310229284.7 Chinese patent embodiment 4 steps 3) product 2, the isocyanate-modified Polyethylene Glycol of 2'-dithio diethyl.

Referring to Fig. 1, the present invention is by S, two (the α of S'-, α '-dimethyl-α " acetic acid) trithiocarbonate is as reversible addition-fracture chain transfer polymerization (RAFT) chain-transferring agent, continuous two steps are implemented N-tertbutyloxycarbonyl propylene hydrazides, the RAFT polymerization of N-(3-dimethylamine propyl) Methacrylamide and N hydroxymethyl acrylamide, then utilize in block copolymer hydroxyl on N hydroxymethyl acrylamide to react with isocyanation esterification end methoxy poly (ethylene glycol), realization connects Polyethylene Glycol through disulfide bond, after deprotection base, obtain required hydrophilic copolymers, gather (N-(3-dimethylamine propyl) Methacrylamide)-polyacrylic hydrazide-Polyethylene Glycol block or comb-shaped copolymer.Amycin is a kind of broad-spectrum anti-cancer drug, and it suppresses nucleic acid synthetic by intercalation of DNA double-spiral structure, realizes antitumor action.Itself presents hydrophobicity amycin, the water miscible hydrochloride form of many employings is with the medication of intravenous drip mode clinically, amycin can be distributed in rapidly whole body like this, toxic and side effects is larger, as there will be bone marrow transplantation, alopecia, the problem such as keep in dark place, and amycin Half-life in vivo is short, and bioavailability is low, and curative effect is not ideal enough.For improving curative effect, the present invention has prepared the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive, this carrier is become amphipathic after loading amycin from hydrophilic, can self assembly form nanoparticle, then through blood circulation or topical mode, can effectively improve drug effect and reduce toxic and side effects.

With respect to traditional free radical polymerisation process, the RAFT polymerization that the present invention adopts can effectively be controlled molecular weight, both can guarantee that synthetic cationic polymer carrier had high-efficiency transfection effect, again can restriction molecule amount in certain limit, guarantee that these not biodegradable acrylamide/acrylic polymers can excrete by body Excretory system, not accumulation in vivo.

Claims

1. a synthetic method for the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive, is characterized in that comprising the steps:

1) N-tertbutyloxycarbonyl propylene hydrazides is synthetic:

2. hydrophilic copolymers pharmaceutical carrier and the synthetic method of pH/ according to claim 1 reduction Dual Sensitive, it is characterized in that: described step 1) solvent in the organic solution of acryloyl chloride is chloroform, dichloromethane or dioxane, step 2) and step 3) in anhydrous and oxygen-free organic solvent be anhydrous and oxygen-free dioxane, anhydrous and oxygen-free chloroform or anhydrous and oxygen-free oxolane; Step 4) anhydrous organic solvent in is anhydrous dimethyl formamide, anhydrous dioxane, anhydrous chloroform, dry toluene or anhydrous tetrahydro furan.

3. the synthetic method of the hydrophilic copolymers pharmaceutical carrier of pH/ according to claim 1 reduction Dual Sensitive, is characterized in that: described step 2) and step 3) in initiator be azodiisobutyronitrile or 4,4'-azo pair (cyanopentanoic acid).

4. the synthetic method of the hydrophilic copolymers pharmaceutical carrier of pH/ according to claim 1 reduction Dual Sensitive, it is characterized in that: S described step 2), two (the α of S'-, α '-dimethyl-α " acetic acid) mol ratio of trithiocarbonate and initiator is 5:1, step 3) and in the mol ratio of poly-(N-tertbutyloxycarbonyl propylene hydrazides) macromole RAFT polymerizable chain transfer agents and initiator be 10:1.

5. the synthetic method of the hydrophilic copolymers pharmaceutical carrier of pH/ according to claim 1 reduction Dual Sensitive, it is characterized in that, poly-(the N-tertbutyloxycarbonyl propylene hydrazides) that described process dewaters-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer is adopted dehydration with the following method and is obtained: in poly-to removing (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide) copolymer, add toluene, under nitrogen atmosphere, reflux and dewater 2～6 hours.

6. the synthetic method of the hydrophilic copolymers pharmaceutical carrier of pH/ according to claim 1 reduction Dual Sensitive, it is characterized in that, described 2, the isocyanate-modified end methoxy poly (ethylene glycol) of 2'-dithio diethyl is adopted with the following method and is obtained: the end methoxy poly (ethylene glycol) that is 800～4000Da by molecular weight, dibutyl tin laurate and 2, 2'-dithio diethyl isocyanates is dissolved in dry toluene, under 85 ℃ of blanket of nitrogen, react 48 hours, then with anhydrous n-hexane, be settled out solid, collect solid drying, obtain 2, the end methoxy poly (ethylene glycol) that 2'-dithio diethyl is isocyanate-modified, wherein, the mol ratio of end methoxy poly (ethylene glycol), dibutyl tin laurate and 2,2'-dithio diethyl isocyanates is 1:(0.02～0.08): (3～8).

7. the synthetic method of the hydrophilic copolymers pharmaceutical carrier of pH/ according to claim 6 reduction Dual Sensitive, it is characterized in that: described step 5) adopt and remove with the following method tertbutyloxycarbonyl protecting group: will gather (N-tertbutyloxycarbonyl propylene hydrazides)-gather (N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer and be dissolved in the mixed solvent that the trifluoroacetic acid of 0 ℃ and dichloromethane form, again in room temperature reaction 4～10 hours, reaction finishes the rear concentrated reaction system obtaining, then with sodium bicarbonate solution, neutralize, then in the ammonia that utilizes dialyzer that molecular cut off is 3500Da to be 0.25% in mass concentration, dialyse, last lyophilizing, obtain the hydrophilic copolymers pharmaceutical carrier of pH/ reduction Dual Sensitive, wherein, the trifluoroacetic acid in mixed solution is (15～30) with the mol ratio of gathering the tertbutyloxycarbonyl in (N-tertbutyloxycarbonyl propylene hydrazides)-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer: 1.

8. a hydrophilic copolymers pharmaceutical carrier that adopts pH/ that in claim 1～7, the synthetic method described in any one claim makes reduction Dual Sensitive, it is characterized in that: its chemical name is polyacrylic hydrazide-poly-(N-(3-dimethylamine propyl) acrylamide/N hydroxymethyl acrylamide)-ethylene glycol copolymer, and chemical structural formula is:

9. the application of the hydrophilic copolymers pharmaceutical carrier of a pH/ reduction Dual Sensitive as claimed in claim 8 in loading amycin and genomic medicine.

10. application according to claim 9, is characterized in that: described genomic medicine is siRNA.