CN115068448B - Single-hole hollow nano-drug carrier with controllable morphology and preparation method and application thereof - Google Patents

Single-hole hollow nano-drug carrier with controllable morphology and preparation method and application thereof Download PDF

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CN115068448B
CN115068448B CN202210702307.0A CN202210702307A CN115068448B CN 115068448 B CN115068448 B CN 115068448B CN 202210702307 A CN202210702307 A CN 202210702307A CN 115068448 B CN115068448 B CN 115068448B
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张明翠
杨倩倩
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Anhui Normal University
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Abstract

The invention provides a morphology-controllable single-hole hollow nano-drug carrier, a preparation method and application thereof, and a high polymer PSI oAm‑NAPI Dissolving in good solvent to obtain organic phase; sodium hydroxide solution is used as the aqueous phase, and the two-phase solution is injected into the reaction chamber at a high speed at the same time, so that the two-phase solution is rapidly and microscopically mixed, turbulence is generated at the interface of the two phases, and the hydrophilic organic solvent is caused to spontaneously diffuse from the oil phase to the aqueous phase. Meanwhile, due to high-speed injection, the pressure gradient generated by the two-phase interface forces the hydrophilic nanoshell to generate a single hole, and the thinner one-side membrane is broken, so that the single hole hollow nano-drug carrier is formed. The single-hole hollow nano-drug carrier is hydrophobic inside and hydrophilic outside, and a definite opening is arranged on the hydrophilic nano-shell. The single-hole hollow nano-drug carrier provided by the invention can be used as an intelligent nano-drug carrier, is convenient for drug encapsulation and delivery, realizes controllable release of anticancer drugs, and has great application potential in biomedical aspect.

Description

Single-hole hollow nano-drug carrier with controllable morphology and preparation method and application thereof
Technical Field
The invention belongs to a material preparation method, and relates to preparation of a novel single-hole hollow nano-drug carrier. In particular to a single-hole hollow nano-drug carrier with controllable morphology, a preparation method and application thereof.
Background
Cancer is the leading cause of death in people, with more than 1000 tens of thousands of people diagnosed each year. At present, early identification and treatment of cancer remains a technical bottleneck, and despite many advances made in conventional therapies such as chemotherapy and radiotherapy, cancer treatment is still far from optimal because of its drawbacks. Such as nonspecific systemic distribution of antitumor drugs, insufficient drug concentration at the tumor site, intolerable cytotoxicity, limited ability to monitor therapeutic response, and development of multidrug resistance. Although several ligand-targeted therapeutic strategies, including immunotoxins, radioimmunotherapy, and drug immunoconjugates, can overcome the problems associated with conventional chemotherapeutic drugs, the limitations of their administration remain a major problem.
Nano-medicine is being explored in depth as an emerging strategy. Nanodrug carriers designed by nanotechnology have many advantages that traditional drug carriers do not possess. The nano-drug carrier can selectively transfer the reporter molecule, the anticancer drug or the gene to tumor tissues through Enhanced Permeability and Retention (EPR) effect, and has the advantages of early effective diagnosis and improved therapeutic effect. The nano-drug carrier can avoid macrophage recognition, prolong blood circulation, and realize excellent treatment effect while reducing side effects. Currently, various types of nano-drug carriers such as polymer micelles, liposomes, microcapsules and the like have been applied to clinical trials, but research literature has found that some problems remain to be solved, such as instability of nanoparticles in blood circulation, low clearance rate of organisms to the nanoparticles, uncontrollable drug release and the like. The unique and diverse properties of its morphology as a nano-drug carrier are advantageous in terms of biocompatibility, pharmacokinetics, targeting and controlled release. Thus, the smart design of a nano-drug carrier system may be more advantageous for drug delivery, and development of nano-drug carriers with unique composition, morphology and surface properties is critical.
In recent years, single-pore nano materials with special structures have attracted wide attention due to the importance of surface opening, and have certain advantages in the aspect of hydrophobic anticancer drug transportation and delivery. However, the morphology of such nano-drug carriers reported in the literature is completely closed shell, and the report of the hollow shape of a single hole is almost blank.
Disclosure of Invention
The invention aims to provide a single-hole hollow nano-drug carrier with controllable morphology and a preparation method thereof, wherein the single-hole hollow nano-drug carrier prepared by a rapid nano-precipitation method has amphipathy, the outside of the single-hole hollow nano-drug carrier is composed of carboxyl with hydrophilicity and N- (3-aminopropyl) imidazole, and the inside of the single-hole hollow nano-drug carrier is formed into a hydrophobic cavity by an oleylamine chain. The particle size of the single-pore nano-drug carrier is 100-500 nanometers, and the pore diameter is 50-100 nanometers. The synthesis method is simple, quick and low in cost, and is suitable for modern chemical preparation.
The invention provides an application of a shape-controllable single-hole hollow nano-drug carrier, which is used for carrying drugs.
The specific technical scheme of the invention is as follows:
the preparation method of the morphology-controllable single-hole hollow nano-drug carrier specifically comprises the following steps:
the high molecular polymer PSI OAm-NAPI Dissolving in good solvent to obtain organic phase, adding aqueous solution of sodium hydroxide or mixed solution of aqueous solution of sodium hydroxide and absolute ethanol to obtain aqueous phase, injecting the two-phase solution into reaction chamber of double-flow limited impact jet mixer, collecting nano precipitation suspension at outlet end, standing, centrifuging, and washing with water to obtain the final product.
The high molecular polymer PSI OAm-NAPI Is prepared by grafting oleylamine and N- (3-aminopropyl) imidazole to polysuccinimide; the preparation method comprises the following steps: heating and reacting Polysuccinimide (PSI) with molecular weight of 6000, oleylamine (OAm) and N- (3-aminopropyl) imidazole (NAPI) in N, N-Dimethylformamide (DMF) solvent, removing impurity DMF by rotary evaporation with absolute ethanol, precipitating with methanol, centrifuging, and drying to obtain polymer grafted polymer PSI OAm-NAPI . The polysuccinimide with the molecular weight of 6000 is adopted in the preparation process, so that the polysuccinimide has low viscosity, is favorable for degradation of the polymer, and has good crystallization rate.
The high molecular polymer PSI OAm-NAPI In the preparation method of (2), the mass ratio of polysuccinimide to the volume ratio of N, N-dimethylformamide is as follows: 0.05:1g/ml; the dosage ratio of the mass of the polysuccinimide to the volume of the oleylamine is 1.6:1.63g/ml; the mass ratio of the polysuccinimide to the oleylamine N- (3-aminopropyl) is as follows: 1:0.0775; PSI used in the preparation of the single pore material of the present invention OAm-NAPI When the NAPI dosage meets the conditions, the single-hole hollow nano-drug carrier with controllable morphology can be prepared only, otherwise, the morphology material of the invention can not be obtained.
The high molecular polymer PSI OAm-NAPI The heating reaction in the preparation method refers to: reacting for 3-7 hours at 60-100 ℃;
in the preparation method of the morphology-controllable single-pore hollow nano-drug carrier, the good solvent is any one of analysis-grade tetrahydrofuran (AR-THF), chromatographic-grade tetrahydrofuran (HPLC-THF), dimethyl sulfoxide (DMSO) or N, N-Dimethylformamide (DMF); analytical grade tetrahydrofuran (AR-THF) is preferred.
The high molecular polymer PSI OAm-NAPI Dissolving the nano particles into a good solvent, wherein the concentration is 6-9mg/mL, and if the concentration is too low, the yield of the obtained nano particles is low, and the morphology of the particles is poor; if the concentration is too high, the size of the obtained nanoparticle is large. The nanometer particles prepared in the proper concentration range of 6-9mg/mL have good appearance and uniform particle size.
The concentration of the sodium hydroxide aqueous solution is 2 multiplied by 10 -5 g/mL。
When the water phase is a mixed solution of sodium hydroxide aqueous solution and absolute ethyl alcohol, the volume of the absolute ethyl alcohol is less than or equal to 1/5 of the volume of the sodium hydroxide aqueous solution.
The volume ratio of the organic phase to the aqueous phase is as follows: 1:1.
in the preparation method of the morphology-controllable single-hole hollow nano-drug carrier, an organic phase and a water phase are injected into a reaction chamber, the mixing time of the two-phase solution is 2-3 seconds, and the Reynolds number is 5000-10000, so that the Re is rapidly and microscopically mixed; in the preparation method of the morphology-controllable single-hole hollow nano-drug carrier, standing is carried out for 10-30min; the centrifugation is carried out at 8000-10000rpm for 8-10min;
the dual-flow restricted impingement jet mixer is referred to as a CIJ-D dual port mixer with two fluid injection ports and one outlet port. The two fluid inlets are an organic phase inlet and an aqueous phase inlet, respectively.
Preferably, the reynolds number of the fluid flow is re=9437. The Reynolds number can judge the fluid flow state, re <2000, is generally laminar flow; re >4000, is generally turbulent. The higher the reynolds number, the more significant the fluid flow state is affected by inertial forces, i.e. the nano-drug carrier is prepared at high speed.
The invention adopts the mixer to prepare nano particles, two phases are mixed with equal momentum, otherwise, the mixing effect is affected, and therefore, the volume ratio of the water phase to the organic phase is 1:1.
in the preparation method, in the rapid mixing process, an amide bond of polysuccinimide is hydrolyzed to form carboxyl under alkaline condition to become hydrophilic group; the grafted oleylamine has hydrophobicity. Because the whole reaction system is carried out in a weakly alkaline aqueous solution (the mixer is used for preparing the nano particles and is provided with two fluid injection ports, wherein one fluid is a 0.5mmoL/L sodium hydroxide aqueous solution, namely, the weakly alkaline aqueous solution is provided for the reaction system), hydrophobic oleylamine chains are far away from the aqueous solution and are close to each other to form a core through hydrophobic interaction, and hydrophilic parts are distributed in the aqueous solution, so that a layer of oil-in-water spherical structure is formed, namely, the inside is a hydrophobic core, and the outside is a hydrophilic shell. In the high-speed injection process, the external pressure forces the hydrophilic nanometer shell to generate a hole, the thinner side film is broken, and the nanospheres are in an equilibrium state, so that the single-hole hollow nanometer drug carrier PSI with a special structure is formed OAm-NAPI 。
The single-hole hollow nano-drug carrier with controllable morphology is prepared by the method, the particle size of the single-hole hollow nano-drug carrier with controllable morphology is distributed at 100-500nm, and the pore size is distributed at 50-100nm. By varying the aqueous phase of absolute ethanol with aqueous sodium hydroxide (2X 10) -5 g/mL) can realize the size regulation of the aperture. In the mixing process, amide bonds of polysuccinimide are hydrolyzed to form carboxyl groups, an oleylamine chain is hydrophobic, the whole reaction system is carried out in a hydrophilic solution, the oleylamine chains are close to each other through hydrophobic interaction to avoid boiled water, so that a hydrophobic core is formed, namely, an inner hydrophobic cavity is formed, and an outer hydrophilic shell is formed. The single-hole hollow structure nano-drug carrier is characterized in that the inside of the nano-drug carrier is a hydrophobic cavity formed by an oleylamine (OAm) chain, the outside of the nano-drug carrier is a hydrophilic shell formed by an amide bond hydrolyzed carboxylate and an N- (3-aminopropyl) imidazole (NAPI) grafted polysuccinimide main chain, and the nano-drug carrier has a definite single hole on the hydrophilic shell of the nano-sphere.
The invention provides an application of a shape-controllable single-hole hollow nano-drug carrier, which is used for carrying drugs, in particular hydrophobic drugs. Can be used for loading and controllable releasing of hydrophobic anticancer drugs. Compared with a nano-drug carrier with a complete closed shell, the single-hole nano-drug carrier is beneficial to the controllable release of hydrophobic anticancer drugs; compared with a porous nano-drug carrier, a single-pore nano-drug carrier can avoid a great deal of leakage of drugs during transportation.
The specific application method comprises the following steps: hydrophobic drugs and high molecular polymers PSI oAm-NAPI Dissolved in a good solvent to serve as an organic phase; aqueous sodium hydroxide solution was used as the aqueous phase, and the two-phase solution was injected simultaneously into the reaction chamber of a dual-flow restricted impact jet mixer, and the nano-drug suspension was collected at the outlet end and dialyzed to remove the free drug. The hydrophobic drug is entrapped in the hydrophobic core by hydrophobic interactions. The dialyzed nano-drug particles were then dispersed in aqueous BSA solution and gently stirred for 12h, and BSA was coated on the nano-particles by electrostatic adsorption. And finally, centrifugally collecting.
In the specific application, the preparation conditions and parameter control are the same as those in the preparation method of the single-hole hollow nano-drug carrier with controllable morphology; the dosage ratio of the hydrophobic drug to the high molecular graft polymer is 1:13.5; the specification of the dialysis bag is MWCO 3500D, and the diameter is 22mm; the dialysis time used was 48 hours; the dialysate used was deionized water. The concentration of the aqueous BSA solution used was 1mg/mL; the speed of the gentle stirring is in the range of 100rpm-300rpm; the centrifugation speed and time were 10000rpm,10min.
The nano-drug carrier prepared by the invention can release drugs in the environment with the pH value of 4.5-5.5, and has the best release performance under the condition with the pH value of=5. (pH of cancer cells was 4.5)<pH<5.5). In addition, the spherical wall of the nano-drug carrier is provided with a definite single hole, so that the nano-drug carrier can be used for packaging hydrophobic anticancer drugs, and the BSA nano-valve is used for packaging the nano-drug carrier, so that drug leakage and other protein adsorption during transportation can be prevented. After the nano-drug is endocytosed by cancer cells, the BSA nano-valve is phosphorylated to hydrolyze, and finally the drug is released continuously. The invention prepares the novel single-hole hollow nano-drug carrier (PSI) of oleylamine and N- (3-aminopropyl) imidazole grafted polysuccinimide by a rapid nano-precipitation method OAm-NAPI ) The method is simple, quick and low in cost. Oleylamine (OAm) and N- (3-aminopropyl)) Imidazole (NAPI) grafted Polysuccinimide (PSI) high molecular polymer PSI oAm-NAPI Dissolved in a good solvent to serve as an organic phase; an aqueous sodium hydroxide solution of low concentration was used as the aqueous phase. The two-phase solution is injected into the reaction chamber at a high speed at the same time, so that the two-phase solution is rapidly and microscopically mixed, turbulence is generated at the interface of the two phases, and the hydrophilic organic solvent is spontaneously diffused from the oil phase to the water phase. Meanwhile, due to high-speed injection, the pressure gradient generated by the two-phase interface forces the hydrophilic nanoshell to generate a single hole, and the thinner one-side membrane is broken, so that the single hole hollow nano-drug carrier is formed. The single-hole hollow nano-drug carrier provided by the invention can be used as an intelligent nano-drug carrier, is convenient for drug encapsulation and delivery, realizes controllable release of anticancer drugs, and has great application potential in biomedical aspect. And the polysuccinimide is used as one of the most promising nano drug carrier materials, has excellent biocompatibility, degradability, high drug loading capacity and high stability, has excellent reactivity with various monomers, and can be introduced by grafting. Polymer nano-drug carrier (PSI) of oleylamine and N- (3-aminopropyl) imidazole grafted polysuccinimide oAm-NAPI ) Is an intelligent drug delivery system responding to pH, and the hydroxyl group of hydrophobic anticancer drugs can be combined with PSI oAm-NAPI Hydrogen bonds are formed between the hydrophilic backbones (PSI) and at lower pH, the hydrogen bonds are broken down and the drug released. The polymer nano-drug carrier has the capabilities of improving the solubility of hydrophobic drugs, prolonging the half-life period of the drugs, reducing side effects and improving the therapeutic effect. In addition, compared with the nano-drug carrier with a complete closed shell, the single-hole hollow nano-drug carrier is beneficial to the encapsulation and release of effective load; compared with a nano-drug carrier with a porous structure, the single-pore nano-drug carrier can reduce a great amount of leakage of drugs in the process of transportation; in addition, the single-hole nano-drug carrier has important significance in the aspect of drug controlled release kinetics, can provide sustained and controllable release of drugs, and is beneficial to reducing drug toxicity and improving drug treatment effect. In summary, single pore nanoparticles have high efficacy as drug carriers based on structural and textural characteristics.
Drawings
FIG. 1 is a schematic diagram of a conventional gas turbineThe single-hole hollow nano-drug carrier PSI of the material D prepared by the method of the invention OAm-NAPI 20 x SEM images of (a);
FIG. 2 is a single-pore hollow nano-drug carrier PSI of material D prepared by the method of the invention OAm-NAPI 40 x SEM images of (a);
FIG. 3 is a single-pore hollow nano-drug carrier PSI of material D prepared by the method of the present invention OAm-NAPI 80 x SEM images of (a);
FIG. 4 is a single-pore hollow nano-drug carrier PSI of material D prepared by the method of the present invention OAm-NAPI A TEM image of (a);
FIG. 5 is a schematic illustration of the preparation of the present invention;
FIG. 6 is an SEM image of material A;
FIG. 7 is an SEM image of material B;
FIG. 8 is an SEM image of material C;
fig. 9 shows that the volume ratio of sodium hydroxide solution to absolute ethanol in the aqueous phase is 11: 1;
fig. 10 shows a volume ratio of sodium hydroxide solution to absolute ethanol in the aqueous phase of 5: 1;
fig. 11 shows a volume ratio of sodium hydroxide solution to absolute ethanol in the aqueous phase of 2: 1;
fig. 12 shows that the volume ratio of sodium hydroxide solution to absolute ethanol in the aqueous phase is 1:1;
FIG. 13 is an SEM image of the post-drug-loaded particles;
FIG. 14 is a fluorescence spectrum demonstrating successful drug loading.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions in the embodiments of the present invention will be clearly and completely described in the following in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The test materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The Polysuccinimide (PSI) used in the invention is purchased from Desai chemical industry Co., ltd, oleylamine, N- (3-aminopropyl) imidazole (NAPI) from Aba Ding Shiji (Shanghai) Co., ltd, N-Dimethylformamide (DMF) from Shanghai Biotechnology Co., ltd, analytical grade tetrahydrofuran (AR-THF), absolute ethyl alcohol, absolute methyl alcohol, sodium hydroxide from national pharmaceutical Congress chemical reagent Co., ltd, bovine Serum Albumin (BSA) from Homeow Biotechnology Co., ltd, and Camptothecine (CPT) from Shanghai Michael Biotechnology Co., ltd.
Those of skill in the art, without any particular mention of the techniques or conditions, may follow the techniques or conditions described in the literature in this field or follow the product specifications.
Example 1
The preparation method of the morphology-controllable single-hole hollow nano-drug carrier comprises the following steps:
1) Polymer graft polymer PSI OAm-NAPI Is prepared from the following steps:
1-1) 1.6g of PSI was dissolved in 32mL of N, N-dimethylformamide in a round bottom flask as a reaction vessel under constant temperature magnetic stirring at 60℃and after the solution had been made to a reddish wine, 1.63mL of oleylamine was added thereto, at which point the temperature was set to 90℃and after 10 minutes of reaction, 0.871g,0.622g,0.373g and 0.124g of N- (3-aminopropyl) imidazole (NAPI) were added, respectively, followed by reaction at 100℃for 5 hours. After the reaction was completed, the mixture was cooled to room temperature.
1-2) to a round bottom flask was added 2/3 of the volume of absolute ethanol solution, and the impurity DMF was removed by rotary evaporation at 80℃and the procedure was repeated 4 times.
1-3) the solution was added dropwise to 280mL of anhydrous methanol solution, allowed to precipitate out, and centrifuged at 10000rpm/10 min. Collecting precipitate, drying in oven at 50deg.C to obtain polymer PSI OAm-NAPI 。
The obtained polymer PSI OAm-NAPI Respectively designated material a, material B, material C and material D. Wherein material A is prepared by adding 0.871g NAPIMaterial B was obtained by adding 0.622g of NAPI, material C was obtained by adding 0.373g of NAPI, and material D was obtained by adding 0.124g of NAPI, and stored at room temperature.
2) Dissolving 27mg of material D in 3ml of analysis-grade tetrahydrofuran by using an ultrasonic instrument with the power of 70W until the material D is completely dissolved for 3min; obtaining clear and transparent solution as an organic phase; another 3mL of 2X 10 -5 As an aqueous phase, g/mL (i.e., 0.5 mmol/L) of an aqueous sodium hydroxide solution was used. The mixer base is stabilized by the left hand, the two-phase solution is injected into the reaction chamber of the double-flow limited impact jet mixer at high speed by the palm center of the right hand, the Reynolds number is Re=9437, the mixing time of the two-phase solution is 2-3 seconds, the two-phase solution is subjected to rapid micromixing, the nano precipitation suspension is collected at the outlet end of the device, after standing for 20min, the nano precipitation suspension is centrifuged at 10000rpm for 10min, washing and sample dripping are performed, characterization is performed, and the nano particles prepared by the material D have good morphology and single Kong Qingxi on the spherical wall are visible. The SEM image and TEM image of the obtained single-hole hollow nano-drug carrier are shown in fig. 1, fig. 2, fig. 3 and fig. 4, the particle size of the single-hole nano-drug carrier is distributed at 100-500 nanometers, and the pore size is distributed at 50-100 nanometers. Due to the angle problem, some holes are on the back side.
Comparative example 1
Taking 27mg of the prepared material A in 3mL of analysis-grade tetrahydrofuran, and performing ultrasonic dissolution by using an ultrasonic instrument with the power of 70W until the material A is completely dissolved, so as to obtain clear and transparent solution as an organic phase; another 3mL of 2X 10 -5 As an aqueous phase, g/mL of an aqueous sodium hydroxide solution was used. Product characterization was obtained in the same manner as in example 1, see fig. 6.
Comparative example 2
Taking 27mg of the prepared material B in 3mL of analysis-grade tetrahydrofuran, and performing ultrasonic dissolution by using an ultrasonic instrument with the power of 70W until the material B is completely dissolved to obtain a clear and transparent solution as an organic phase; another 3mL of 2X 10 -5 As an aqueous phase, g/mL of an aqueous sodium hydroxide solution was used. Product characterization was obtained in the same manner as in example 1, see fig. 7.
Comparative example 3
27mg of material C was taken in 3mL analytical grade tetrahydroIn furan, using an ultrasonic instrument with the power of 70W to carry out ultrasonic dissolution until the material C is completely dissolved, so as to obtain clear and transparent solution as an organic phase; another 3mL of 2X 10 -5 As an aqueous phase, g/mL of an aqueous sodium hydroxide solution was used. Product characterization was obtained in the same manner as in example 1, see fig. 8.
By comparing the example 1 with the comparative examples 1-3, it is found that the materials A, B and C cannot prepare the single-hole hollow nano-drug carrier with morphology by a rapid nano-precipitation method, and the material D can prepare the single-hole hollow nano-drug carrier with good morphology and distinct single holes. Because of the poor solubility of materials A, B, C in tetrahydrofuran, long-time ultrasound is required to obtain clear and transparent solutions. The material A has the worst solubility, the material B has the inferior solubility, the material C has better solubility, the ultrasound lasts for about 40 minutes, and the ultrasound lasts for about 20 minutes, and in the process, the polymer PSI is grafted by the polymer oAm-NAPI And the particles with morphology can be damaged and are difficult to prepare through a rapid nano precipitation method. The material D with NAPI content of 0.124g has better solubility in tetrahydrofuran, can be dissolved only by ultrasonic treatment for 3 minutes, can prepare nano particles with good morphology by a rapid nano precipitation method, and has a single-hole hollow structure.
Example 2
The preparation method of the morphology-controllable single-hole hollow nano-drug carrier comprises the following steps:
taking 27mg of the prepared material D in 3ml of analysis-grade tetrahydrofuran, and performing ultrasonic dissolution by using an ultrasonic instrument with the power of 70W until the material D is completely dissolved, so as to obtain a clear and transparent solution as an organic phase; the water phase was changed to 2X 10 -5 The volume ratio of the mixed solution of the sodium hydroxide aqueous solution and the absolute ethyl alcohol of g/mL is changed to 11:1,5:1,2:1,1:1; the total volume of the aqueous phase was still 3mL. According to the characterization, the single pore diameter of the nano particles is reduced or even eliminated with the increase of the absolute ethanol content, and the SEM images are shown in figures 9-12.
As can be seen from a combination of examples 1 and 2, the aqueous phase is pure sodium hydroxide solution or the aqueous phase is a volume ratio of sodium hydroxide solution to absolute ethanol of 11:1,5:1, a nano-drug carrier with a single-hole hollow structure can be realized, but according to a scanning diagram, the prepared nano-particle single-hole hollow structure has better morphology under the condition of no absolute ethyl alcohol.
Example 3
The application of the morphology-controllable single-hole hollow nano-drug carrier for carrying drugs specifically comprises the following steps:
dissolving 27mg of the prepared material D and 2mg of Camptothecine (CPT) together in 3ml of analysis-grade tetrahydrofuran, and performing ultrasonic dissolution by using an ultrasonic instrument with the power of 70W to obtain a clear and transparent solution as an organic phase; another 3mL of 2X 10 -5 A g/mL sodium hydroxide solution was used as the aqueous phase. The two-phase solution is injected simultaneously into the reaction chamber of a dual-flow restricted impingement jet mixer at high speed to allow rapid micromixing and collecting the nano-precipitation suspension at the outlet end of the device. Dialysis was performed for 48 hours using a dialysis bag with a molecular weight cut-off of 3500 daltons and a diameter of 22mm to remove free drug, during which time the dialysate was replaced 8 times with deionized water. Finally dispersing the dialyzed nano-drug particles in 1mg/mL BSA aqueous solution, stirring for 12 hours at a rotating speed of 100rpm, wrapping the BSA on the nano-drug particles in an electrostatic adsorption mode, and centrifuging at 10000rpm for 10 minutes to obtain the nano-drug. The nanoparticle morphology remained stable after dialysis, see SEM fig. 13, upper right inset is TEM image after drug loading. Since camptothecin has a fluorescence emission peak at a wavelength of 430nm, for this purpose, it was characterized using a fluorescence spectrometer, see fig. 14. (CPT is purchased; PSI in the figure) OAm-NAPI Is a single-hole hollow nanoparticle which is prepared by a mixer and is not loaded with medicine; PSI (program specific information) OAm-NAPI CPT-BSA is the fluorescence absorbance of the drug-loaded nanoparticle as measured without destruction; PSI (program specific information) OAm-NAPI CPT-BSA (destroyed) is the fluorescence absorbance measured after drug-loaded nanoparticle destruction. The destruction refers to dispersing the obtained carrier in ethanol solution, and ultrasonic destruction for 3 hours) according to the fluorescence emission peak change before and after the destruction of the drug-loaded nano particles, the drug loading can be proved to be successful. The nano-drug can continuously release about 80% of the drug within 230h under the physiological environment of pH=5.0.

Claims (6)

1. The preparation method of the morphology-controllable single-hole hollow nano-drug carrier is characterized by comprising the following steps of:
the high molecular polymer PSI OAm-NAPI Dissolving in a good solvent to serve as an organic phase, taking a sodium hydroxide solution as a water phase, simultaneously injecting the two-phase solution into a reaction chamber of a double-flow limited impact jet mixer, collecting nano precipitation suspension at an outlet end, standing, centrifuging, and washing with water to obtain a single-hole hollow nano drug carrier with controllable morphology;
the high molecular polymer PSI OAm-NAPI The preparation method of (2) comprises the following steps: heating and reacting Polysuccinimide (PSI) with molecular weight of 6000, oleylamine (OAm) and N- (3-aminopropyl) imidazole (NAPI) in N, N-Dimethylformamide (DMF) solvent, removing impurity DMF by rotary evaporation with absolute ethanol, precipitating with methanol, centrifuging, and drying to obtain polymer grafted polymer PSI OAm-NAPI ；
The mass ratio of the polysuccinimide to the N- (3-aminopropyl) imidazole is as follows: 1:0.0775;
the good solvent is analytical grade tetrahydrofuran;
the reynolds number of the fluid flow is re=9437;
the volume ratio of the organic phase to the aqueous phase is as follows: 1:1, a step of;
the particle size of the single-hole hollow nano-drug carrier with controllable morphology is 100-500 nanometers, and the pore diameter is 50-100 nanometers.
2. The method of claim 1, wherein the high molecular polymer PSI OAm-NAPI Dissolving in good solvent at a concentration of 6-9mg/mL.
3. The method according to claim 1, wherein the concentration of the sodium hydroxide solution is 2X 10 -5 g/mL。
4. The method of claim 1, wherein the organic phase and the aqueous phase are injected into the reaction chamber and the mixing time of the two-phase solution is 2 to 3 seconds.
5. The morphology-controllable single-hole hollow nano-drug carrier prepared by the preparation method of any one of claims 1-4, which is prepared by the method, wherein the particle size of the morphology-controllable single-hole hollow nano-drug carrier is 100-500 nanometers, and the pore size is 50-100 nanometers.
6. Use of a single-pore hollow nano-drug carrier with controllable morphology prepared by the preparation method of any one of claims 1-4 for drug delivery.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008222810A (en) * 2007-03-12 2008-09-25 Sekisui Plastics Co Ltd Single-hole hollow particle and its production method
CN107091826A (en) * 2017-06-24 2017-08-25 安徽师范大学 A kind of method that PSI OAm NAPI amphiphilic polymer/nanometer pharmaceutical carriers are quantitatively detected based on trace fluorescence immunoassay
CN110478496A (en) * 2019-08-28 2019-11-22 安徽师范大学 A kind of preparation method of the controllable polymeric nano medicine carrier oleyl amine scion grafting polysuccinimide of uniform particle sizes, size

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008222810A (en) * 2007-03-12 2008-09-25 Sekisui Plastics Co Ltd Single-hole hollow particle and its production method
CN107091826A (en) * 2017-06-24 2017-08-25 安徽师范大学 A kind of method that PSI OAm NAPI amphiphilic polymer/nanometer pharmaceutical carriers are quantitatively detected based on trace fluorescence immunoassay
CN110478496A (en) * 2019-08-28 2019-11-22 安徽师范大学 A kind of preparation method of the controllable polymeric nano medicine carrier oleyl amine scion grafting polysuccinimide of uniform particle sizes, size

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