CN104771373A - Drug carrying polyalkylcyanoacrylate nanocarrier and preparation method thereof - Google Patents

Abstract

The invention belongs to the field of nano preparations and relates to a drug-loaded polyalkylcyanoacrylate nanocarrier and a preparation method thereof. The drug-loaded polycyanoacrylate nanocarrier includes the following components in terms of mass percentage: 1-10% of Drug, 25-75% stabilizer, 9-50% alkyl cyanoacrylate monomer and 9-30% freeze-drying protectant; the preparation method of the drug-loaded polyalkylcyanoacrylate nanocarrier is firstly adding the drug It is polymerized together with the alkyl cyanoacrylate monomer in a concentrated alcohol system, and then the obtained polymer system is slowly transferred to the water phase and stirred for a certain period of time, and then processed by rotary evaporation, ultrasonication, filtration and freeze-drying. Available as a powder. In the present invention, the polymerization reaction of the alkyl cyanoacrylate monomer is basically carried out in alcohol, which can shorten the contact time between the medicine and the acidic medium and prevent the medicine from degrading.

Description

A drug-loaded polyalkylcyanoacrylate nanocarrier and its preparation method

technical field

The invention relates to the field of nano preparations, in particular to a drug-loaded polyalkylcyanoacrylate nano carrier and a preparation method thereof.

Background technique

The development of drug delivery systems has attracted widespread attention in the search for more effective ways to treat life-threatening diseases in humans and animals. The attractiveness of designing a drug delivery system as a modern treatment method lies in: (1) Adsorption or encapsulation of therapeutic drugs on carriers or excipients can precisely deliver drugs to specific organs or parts, or achieve disease-specific targets; (2) It can control the release of drugs and prolong the half-life; (3) Provide new or more convenient routes of administration, so that drugs can reach parts of the body that are difficult to transport; (4) It can better improve drug-related parameters, such as Pharmacokinetics, pharmacodynamics, non-specific toxicity, immunogenicity, etc., to improve therapeutic efficacy (Vanderwoot, J., Ludwig, A. Ocular drug delivery: nanomedicines application [J]. Nanomedicine, 2007, 2: 11-21. ). Most drug delivery systems are based on nanoparticles, including nanoparticles, liposomes, emulsions, resinous polymers, carbon nanotubes, etc.

Many synthetic or natural polymers have been used in drug transport studies. Although natural polymers such as collagen and cellulose can be used to deliver various protein drugs, these materials tend to degrade slowly, even though the performance can be improved by adding various functional groups, such as cross-linking with glutaraldehyde. polymers are degradable, but the degradation products of these modified natural polymers are not desirable in vivo. While poly(alkylcyanoacrylate) polymers undergo enzymatic ester hydrolysis to obtain primary alcohols, butanol and water-soluble poly(2-cyanoacrylic acid) acids. At the same time, during the degradation process of poly(alkyl cyanoacrylate) polymers, the polymer chain remains intact and gradually becomes more hydrophilic until it becomes water-soluble (C.O'Sullivan, C.Birkinshaw.mydrolysis of poly(n-butylcyanoacrylate) nanoparticles using esterase [J]. Polymer Degradation and Stability, 2002, 78:7-15).

Common preparation methods of polyalkylcyanoacrylate nanoparticles are emulsion polymerization and interfacial polycondensation. Emulsion polymerization is suitable for drugs with high solubility and stability in acidic media. In the acidic aqueous solution of the stabilizer, the alkyl cyanoacrylate monomer polymerizes inside the emulsion droplets or micelles formed by the emulsifier (Joshi SA, Chachan SS, Sawant KK. Rivastigmine-loaded PLGA and PBCA nanoparticles: preparation, optimization, characterization, in vitro and pharmacodynamic studies[J].Eur J PharmBiopharm,2010,76:189-99); while the interfacial polycondensation method is suitable for fat-soluble drugs, which is to dissolve alkyl cyanoacrylate monomers and drugs in organic solvents The obtained oil phase is slowly added dropwise to a large amount of water phase containing a stabilizer, and the alkyl cyanoacrylate monomer is polymerized on the two-phase diffusion interface (Fei Ren, Ruda Chen, Ying Wang, et al.Paclitaxel-LoadedPoly (n-butylcyanoacrylate) Nanoparticle Delivery System to Overcome multidrug Resistance in Ovarian Cancer[J].Pharm Res,2011,28:897-906). In these two methods, in order to prevent the rapid polymerization of monomers caused by too much OH- in water, it is generally necessary to adjust the water phase to acidity. In order to ensure that the monomers are fully polymerized, the reaction time is generally not less than 4 hours, which may affect the acidic conditions. Stability of easily hydrolyzed drugs.

Contents of the invention

The purpose of the present invention is to provide a drug-loaded polyalkylcyanoacrylate nano-carrier and its preparation method. The drug and alkylcyanoacrylate monomers are placed in a concentrated alcohol system for polymerization in advance, and then the obtained preliminary The polymer system is slowly transferred dropwise to the water phase and stirred for a certain period of time, and then undergoes operations such as rotary evaporation, ultrasonication, filtration, and freeze-drying. The polymerization reaction of the alkyl cyanoacrylate monomer in the present invention is basically carried out in alcohol, which can shorten The contact time between the drug and the acidic medium reduces the degradation of the drug and enhances the stability of the drug.

The technical solution of the present invention is: a drug-loaded polyalkylcyanoacrylate nanoparticle carrier, which includes the following components in terms of mass percentage: 1-10% of drug, 25-75% of stabilizer, 9-50% of Alkyl cyanoacrylate monomer and 9-30% lyoprotectant;

In the above scheme, the drugs are: bufalin, mitoxantrone, curcumin, paclitaxel, etoposide, paclitaxel, gefitinib, hydroxycamptothecin, doxorubicin, fluorouracil, cyclophosphamide , irinotecan or cisplatin.

The stabilizer is: Poloxamer 188, Poloxamer 407, Pluronic F127, Dextran 70, Dextran 40, Dextran 20, polyvinyl alcohol series, Tween-80, any one of Span-80 or any mixture of several.

The alkyl cyanoacrylate is: methyl cyanoacrylate (MCA), ethyl ester (ECA), butyl ester (BCA), isobutyl ester (IBCA), hexyl ester (HCA) or isohexyl ester (IHCA) any kind.

The lyoprotectant is any one or a mixture of any of the following:

(1) Sugars/polyols: sucrose, trehalose, mannitol, lactose, glucose, maltose;

(2) Polymers: PVP, PEG, dextran, albumin;

(3) Amino acids: L-serine, sodium glutamate, alanine, glycine, sarcosine;

(4) Salts: phosphate, acetate, citrate.

In the above solution, the drug-loaded polyalkylcyanoacrylate nanocarrier has an average particle diameter of 20-1000 nm, preferably 50-200 nm.

The preparation method of the drug-loaded polyalkylcyanoacrylate nanocarrier comprises the following steps:

(1) The drug is polymerized together with the alkyl cyanoacrylate monomer in a concentrated alcohol system:

Calculated by mass percentage, weigh 1-10% of the drug, dissolve it in 5 mL of concentrated alcohol system containing 4.5-45% stabilizer or no stabilizer, that is, ethanol-water system (wherein the ethanol concentration is 89-99%, V /V), controlling the rotation speed to 100-1000 rpm, slowly adding 9-50% alkyl cyanoacrylate monomer dropwise into the concentrated alcohol system, stirring at room temperature for 5 hours, to obtain a preliminary polymer system in alcohol;

(2) Preparation of the aqueous phase:

Calculated by mass percentage, weigh 4-50% of the stabilizer and dissolve it in distilled water, or without adding the stabilizer, adjust to a specific pH2-pH7.4, this is the water phase;

(3) Slowly transfer the preliminary polymer system dropwise to the aqueous phase:

The rotational speed of the water phase is controlled at 100-1000 rpm, the preliminary polymer system is added dropwise into the water phase, and the system is uniformly dispersed by using ultrasonic water bath, wherein the ultrasonic temperature of the water bath is 0-50°C, and the power of the ultrasonic water bath is 10~200W;

(4) Rotary evaporation:

After continuing to stir the system obtained in the step (3) for 0.5-2 hours, adjust the pH to neutral, remove ethanol by vacuum rotary evaporation, and the temperature of the rotary evaporation is 0-60°C;

(5) Probe sonication:

Using probe ultrasound to shear the nanoparticles in the system obtained in step (4) to obtain small nanoparticles, wherein the power of the probe ultrasound is 10 to 1000W, and the number of probe ultrasound is 10 to 600 times ;

(6) Filter:

Filtrating through a filter membrane with a pore size of 0.8 μm to obtain a drug-loaded polyalkylcyanoacrylate nanoparticle suspension;

(7) Freeze drying:

Add 9-30% freeze-drying protectant to the drug-loaded polyalkylcyanoacrylate nanoparticle suspension, and freeze-dry for 24 hours to obtain a powder.

In the above solution, the ultrasonic temperature of the water bath is preferably 10-30°C, and the ultrasonic power of the water bath is preferably 80-100W.

In the above solution, the rotary evaporation temperature is preferably 10-30°C.

In the above solution, the ultrasonic power of the probe is preferably 100-400 W, and the number of ultrasonic probes is preferably 50-400.

Beneficial effects: compared with the prior art, the present invention pre-polymerizes the drug and alkyl cyanoacrylate monomer in a concentrated alcohol system, and then transfers to the water phase for fixation. The drug-loaded polycyanoacrylate prepared by the present invention Alkyl acrylate nanocarriers are spherical, with good physical and chemical properties, which can effectively encapsulate hydrophobic drugs, shorten the contact time between drugs and acidic media, and enhance the stability of drugs; The ultrasonic power is preferably 80-100W, the temperature of the rotary evaporation is preferably 10-30°C, the ultrasonic power of the probe is preferably 100-400W, and the number of ultrasonic probes is preferably 50-400. These optimal conditions can maintain the stability of the drug and avoid The high temperature causes the nanoparticles to deteriorate; in order to make them easier to store, a lyoprotectant is added for freeze-drying to form a powder.

Description of drawings

FIG. 1 is a transmission electron micrograph of the polyn-butyl cyanoacrylate nano-carrier loaded with bufalin prepared according to Example 1.

Fig. 2 is an atlas of the self-control group of rabbit ear vein slices prepared according to Example 1 loaded with bufolin poly-n-butyl cyanoacrylate nanocarrier.

Fig. 3 is an atlas of raw materials of rabbit ear vein slices prepared according to Example 1 loaded with bufalin poly-n-butyl cyanoacrylate nanocarrier.

Fig. 4 is an atlas of rabbit ear vein slice preparation group prepared according to Example 1 loaded with bufalin poly-n-butyl cyanoacrylate nanocarrier.

Detailed ways

The present invention will be further described below in conjunction with the embodiments, but not limited to the following embodiments, those skilled in the art may make various adjustments to the present invention, and these adjustments also belong to the scope of the present invention defined by the appended claims.

A preparation method of a drug-loaded polyalkylcyanoacrylate nanocarrier provided by the invention comprises the following steps:

Calculated by mass percentage, weigh 1 to 10% of the hydrophobic drug, dissolve it in 5 mL of concentrated alcohol system containing 4.5 to 45% stabilizer (wherein the concentration of ethanol is 89 to 99%, V/V); the control speed is 100 At ~1000rpm, slowly add 9-50% alkyl cyanoacrylate monomer dropwise into the above concentrated alcohol system, and stir at room temperature for 5 hours to obtain a preliminary polymer system in alcohol. Calculated by mass percentage, weigh 4-50% stabilizer, dissolve it with distilled water, adjust to a specific pH (2-7.4), this is the water phase;

Or calculate by mass percentage, weigh 1-10% hydrophobic drug, dissolve it in 5mL concentrated alcohol system (wherein the ethanol concentration is 89-99%, V/V), control the rotating speed to be 100-1000rpm, slowly turn 9- 50% alkyl cyanoacrylate monomer was added dropwise to the above-mentioned concentrated alcohol system, and stirred at room temperature for 5 hours to obtain a preliminary polymer system in alcohol. Calculated by mass percentage, weigh 4 to 50% of the stabilizer, dissolve it in distilled water, and adjust to a specific pH (2 to 7.4), which is the water phase;

Or calculate by mass percentage, weigh 1-10% hydrophobic drug, dissolve in 5mL of concentrated alcohol system containing 4.5-45% stabilizer (wherein the ethanol concentration is 89-99%, V/V), control the speed of 100-1000 rpm, slowly add 9-50% alkyl cyanoacrylate monomer dropwise into the above-mentioned concentrated alcohol system, and stir at room temperature for 5 hours to obtain a preliminary polymer system in alcohol. Calculated by mass percentage, adjusted to a specific pH (2-7.4), this is the water phase;

The rotational speed of the water phase is controlled at 100-1000 rpm, the preliminary polymer system is added dropwise into the water phase, and the system is uniformly dispersed by using ultrasonic water bath, wherein the ultrasonic temperature of the water bath is 0-50°C, and the power of the ultrasonic water bath is 10-200W; continue stirring for 0.5-2h, adjust pH to neutral, rotary evaporation temperature is 0-60℃, vacuum rotary evaporation to remove ethanol; 10-1000W probe ultrasonic 10-600 times and pass through 0.8μm filter membrane, Add 9-30% freeze-drying protectant and freeze-dry.

Calculated by mass percentage, the total amount of the concentrated alcohol system and the stabilizer in the water phase is 25-75%.

The ultrasonic temperature of the water bath is preferably 10-30°C.

The ultrasonic power of the water bath is preferably 80-100W.

The rotary evaporation temperature is preferably 10-30°C.

The ultrasonic power of the probe is preferably 100-400W.

The number of ultrasounds of the probe is preferably 50-400.

The average particle size of the drug-loaded polyalkylcyanoacrylate nanocarrier is 20-1000 nm, preferably 50-200 nm.

The water phase is adjusted to a specific pH (2-7.4) because: although the alkyl cyanoacrylate monomer in the present invention has basically completed the polymerization in the ethanol system, it may Some monomers still polymerized and formed nanoparticles in the aqueous phase system. For acid-resistant drugs, the pH can be properly adjusted to acidity, which is conducive to the formation of smaller nanoparticles from unpolymerized monomers. For acid-labile drugs, it is not necessary to adjust the pH of the aqueous phase to maintain neutrality.

Example 1: Preparation of bufolin-loaded poly(n-butyl cyanoacrylate) nanocarrier

Precisely weigh 5 mg of bufalin, dissolve it in a concentrated alcohol system (98% ethanol concentration) containing 20 mg of poloxamer 188, accurately absorb 55 mg of n-butyl cyanoacrylate (α-BCA) monomer, and slowly add it dropwise In the concentrated alcohol system, stir while adding dropwise. After stirring for 5 hours, add the preliminary polymer system in the alcohol dropwise to 5 mL of the aqueous phase containing 48 mg of poloxamer 188 (pH 4.0), 20 ° C, 100 W water bath ultrasonic The system is uniformly dispersed; after continuing to stir for 0.5h, adjust the pH to neutral with sodium hydroxide, and remove ethanol by vacuum rotary evaporation at 30°C; 200W probe is ultrasonicated 2×200 times, passed through a 0.8μm filter membrane, and the prepared nanoparticles The solution was milky white.

Add 25 mg dextran 70 and 30 mg trehalose to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, and the particle size is almost unchanged from that before freeze-drying.

Example 2: Preparation of mitoxantrone-loaded polyisobutyl cyanoacrylate nanocarrier

Precisely weigh 15 mg of mitoxantrone, dissolve it in a concentrated alcohol system (89% ethanol concentration) containing 90 mg of Pluronic F127, accurately absorb 20 mg of isobutyl cyanoacrylate (IBCA) monomer, and slowly add the concentrated alcohol drop by drop In the system, stir while adding dropwise. After stirring for 5 hours, add the preliminary polymer system in alcohol dropwise to 5 mL of water phase (pH 2.0) containing 70 mg of poloxamer 188, 0 ° C, 80 W water bath ultrasonic to disperse the system Uniform; continue to stir for 1 hour, adjust the pH to neutral with sodium hydroxide, and remove ethanol by vacuum rotary evaporation at 20°C; 300W probe ultrasonic 2×10 times, pass through a 0.8μm filter membrane, the prepared nanoparticle solution appears blue color opalescence.

Add 20 mg sucrose to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, and the particle size is almost unchanged compared with that before freeze-drying.

Example 3: Preparation of polymethyl cyanoacrylate nanocarriers loaded with paclitaxel

Precisely weigh 10mg of doclitaxel, dissolve it in a concentrated alcohol system (95% ethanol concentration) containing 25mg of poloxamer 407, accurately absorb 80mg of methyl cyanoacrylate (MCA) monomer, and slowly add it to the concentrated alcohol system drop by drop Add it dropwise while stirring, after stirring for 5 hours, add the preliminary polymer system in alcohol dropwise to 5mL water phase containing 100mg dextran 70 (pH 7.4), 50°C, 10W water bath ultrasonic to disperse the system evenly; continue stirring After 0.5 h, ethanol was removed by vacuum rotary evaporation at 60° C.; 100 W probe was sonicated 2×50 times, and passed through a 0.8 μm filter membrane. The prepared nanoparticle solution was milky white.

Add 20mg sucrose, 20mg PEG 400, 15mg mannitol and 15mg glycine to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, the particle size is compared with that before freeze-drying, Almost unchanged.

Example 4: Preparation of etoposide-loaded polyethyl cyanoacrylate nanocarrier

Accurately weigh 24.5 mg of etoposide, dissolve it in a concentrated alcohol system (93% ethanol concentration) containing 110 mg PEG 400, accurately absorb 40 mg of ethyl cyanoacrylate ECA monomer, slowly add it dropwise to the concentrated alcohol system, and drop Stir while adding, and after stirring for 5 hours, add the preliminary polymer in the alcohol dropwise into 5 mL of the water phase containing 20 mg Tween 80 (pH 6.8), at 10 ° C, 200 W water bath ultrasonic to disperse the system evenly; after continuing to stir for 1 hour, Ethanol was removed by vacuum rotary evaporation at 25°C; a 400W probe was sonicated 2×100 times, passed through a 0.8 μm filter membrane, and the prepared nanoparticle solution was milky white.

Add 25 mg of lactose and 25 mg of mannitol to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, and the particle size has almost no change compared with that before freeze-drying.

Example 5: Preparation of doxorubicin-loaded poly(n-butyl cyanoacrylate) nanocarrier

Precisely weigh 10 mg of doxorubicin, dissolve it in a concentrated alcohol system (96% ethanol concentration) containing 80 mg of Tween 80, accurately absorb 60 mg of n-butyl cyanoacrylate (BCA) monomer, and slowly add it dropwise to the concentrated alcohol system , stirring while adding dropwise, after stirring for 5 hours, add the preliminary polymer system in alcohol dropwise to 5mL water phase containing 10mg polyvinyl alcohol 200 (pH 7.0), 18°C, 50W water bath ultrasonic to make the system uniformly dispersed; continue After stirring for 1 h, the ethanol was removed by vacuum rotary evaporation at 0°C; the 10W probe was sonicated 2×600 times, and passed through a 0.8 μm filter membrane, and the prepared nanoparticle solution showed purple opalescence.

Add 25mg PEG400, 10mg trisodium citrate and 15mg trisodium glycinate to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, the particle size is compared with that before freeze-drying, Almost unchanged.

Embodiment 6: Preparation of loaded curcumin isohexyl cyanoacrylate IHCA nanocarrier

Accurately weigh 5 mg of curcumin, dissolve it in a concentrated alcohol system (98% ethanol concentration) containing 50.5 mg of Span 80, accurately absorb 96 mg of isohexyl cyanoacrylate IHCA monomer, and slowly add it dropwise to the concentrated alcohol system, dripping Stir while adding, and after stirring for 5 hours, add the preliminary polymer system in alcohol dropwise to 5 mL of water phase without stabilizer (pH 3.0), at 30°C, 150W water bath ultrasonic to disperse the system evenly; after continuing to stir for 0.5 hours, Adjust to neutrality with sodium hydroxide, and remove ethanol by vacuum rotary evaporation at 50°C; sonicate 2×10 times with a 1000W probe, pass through a 0.8 μm filter membrane, and the prepared nanoparticle solution exhibits orange-yellow opalescence.

Add 25 mg mannitol and 25 mg trehalose to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to re-disperse to form a nanoparticle solution, and the particle size is almost unchanged from that before freeze-drying.

Example 7: Preparation of gefitinib-loaded poly(n-butyl cyanoacrylate) nanocarrier

Precisely weigh 2.5 mg of gefitinib, dissolve it in a concentrated alcohol system (99% ethanol concentration) without stabilizers, accurately absorb 75 mg of n-butyl cyanoacrylate (BCA) monomer, and slowly add it dropwise to the concentrated alcohol system During the process, stir while adding dropwise, after stirring for 5 hours, add the preliminary polymer system in alcohol dropwise to 5 mL of the water phase containing 80 mg of Pluronic F127 (pH 5.0), at 28 ° C, 95 W water bath ultrasonic to disperse the system evenly; After continuing to stir for 0.5h, adjust to neutrality with sodium hydroxide, and remove ethanol by vacuum rotary evaporation at 30°C; 800W probe ultrasonic 2×600 times, pass through a 0.8μm filter membrane, the prepared nanoparticle solution exhibits brown opalescence .

Add 40 mg trehalose to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, and the particle size has almost no change compared with that before freeze-drying.

Example 8: Preparation of hydroxycamptothecin polyisobutyl cyanoacrylate nanocarrier

Accurately weigh 2.3 mg of hydroxycamptothecin, dissolve it in a concentrated alcohol system (99% ethanol concentration) containing 10 mg of dextran 70, accurately absorb 40 mg of isobutyl cyanoacrylate (IBCA) monomer, and slowly add it dropwise to the concentrated alcohol system In the solution, stir while adding dropwise, after stirring for 5 hours, add the preliminary polymer system in alcohol dropwise to 5mL water phase containing 113mg poloxamer 407 (pH4.0), 30℃, 100W water bath ultrasonic to disperse the system Uniform; continue to stir for 0.5h, adjust to neutral with sodium hydroxide, and remove ethanol by vacuum rotary evaporation at 25°C; 600W probe ultrasonic 2×300 times, pass through a 0.8μm filter membrane, the prepared nanoparticle solution is yellow Opalescence.

Add 25mg mannitol, 20mg PVP K30 and 15.5mg alanine to the filtrate, freeze-dry for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, the particle size is compared with that before freeze-drying, Almost unchanged.

Example 9: Preparation of fluorouracil-loaded poly(n-butyl cyanoacrylate) nanocarrier

Precisely weigh 20 mg of fluorouracil, dissolve it in a concentrated alcohol system (97% ethanol concentration) containing 45 mg of Pluronic F127, accurately absorb 130 mg of n-butyl cyanoacrylate (BCA) monomer, and slowly add it dropwise into the concentrated alcohol system, Stir while adding dropwise, and after stirring for 5 hours, add the preliminary polymer system in alcohol dropwise to 5 mL of water phase containing 40 mg of poloxamer 188 (pH6.8), at 28 ° C, 110 W water bath ultrasonic to disperse the system evenly; After continuing to stir for 0.5 h, ethanol was removed by vacuum rotary evaporation at 20° C.; 500 W probe was sonicated 2×500 times, and passed through a 0.8 μm filter membrane. The prepared nanoparticle solution was milky white.

Add 20mg of glucose and 10mg of PVP K30 to the filtrate and lyophilize for 24 hours to obtain a powder, add sterile water or saline to redisperse to form a nanoparticle solution, and the particle size is almost unchanged from that before lyophilization.

Experimental Example 1: Particle Size and Distribution

The particle size distribution of the nano-carriers prepared in Example 1 was measured by using a Malvern laser particle size analyzer (Zetasizer Nano ZS90 type laser particle size analyzer, Malvern, UK), and the results showed that the bufalin polycyanoacrylic acid prepared by the present invention is positive The average particle size of the butyl ester nanocarrier is 172.15±0.13nm, and the polydispersity coefficient is 0.078±0.021.

Experimental example 2: Morphological distribution

Take an appropriate amount of nano-carriers, dilute with distilled water, put a drop on the copper grid covered with carbon support film, after standing for a while, continue to add a drop of 2% phosphotungstic acid for negative staining, and use a transmission electron microscope (Tecnai 12, Philips company , Holland) observed the particle morphology, and the observation results of implementation example 1 are shown in Fig. 1. It can be seen from Figure 1 that the distribution of particles is relatively uniform and has a regular spherical shape.

Experimental Example 3: Stability Investigation

The nanoparticle solution prepared in Example 1 was placed in a refrigerator at 4°C for 72 hours. Visual observation found that the solution had almost no change, no nanoparticle sedimentation, drug precipitation, etc., and the particle size was detected by a Malvern laser particle size analyzer. After the particle solution was left for 72 hours, the particle size did not change significantly.

Experimental Example 4: Hemolytic Experiment

Investigate the hemolytic properties of bufalin raw material drug and bufalin poly-n-butyl cyanoacrylate nanocarrier prepared in Example 1. Take 12 clean test tubes, label them respectively, and add 2.5mL 2% erythrocyte suspension to each of them, of which No. 1 to No. 5 is the raw material drug group (the concentration of bufalin is 4, 8, 12, 16, 20 μg/mL in turn) ; Nos. 6 to 10 are the bufalin PBCA nanoparticle group (the concentration of bufalin contained in order is 4, 8, 12, 16, 20 μg/mL); No. 11 and No. 12 are added with normal saline and distilled water respectively, as a negative control group and the positive control group. The samples of each group were kept warm in a 37°C water bath, and the hemolysis was observed at 0.3, 1, 2, 3, 4, and 6 hours respectively. Operate 3 times in parallel. The experimental results showed that at 0.3 hours, slight hemolysis was observed in tube No. 5 of the raw material drug group and tube No. 10 of the preparation group, hemolysis occurred in the positive control group (tube No. 12), and no hemolysis was found in the other tubes. With the prolongation of time, moderate hemolysis occurred in No. 5 tube of the API group at 1 hour; slight hemolysis occurred in No. 4 tube of the API group at 2 hours; severe hemolysis occurred at 4 hours. In the preparation group, only tube No. 10 had slight hemolysis, and the other tubes had no hemolysis. It can be seen that high-dose bufalin has a certain hemolytic property, but after bufalin is loaded in PBCA nanoparticles, only slight hemolysis occurs in the high-dose group, and the safety is greatly enhanced.

Experimental Example 5: Vascular Stimulation Experiment

Get 6 New Zealand rabbits (male, 2kg), and divide them into two groups, A and B, wherein group A is the raw material drug group, and group B is the bufalin preparation group prepared in Example 1. The dose of 0.25 mL/kg was injected into the right ear margin vein, and at the same time, each rabbit was injected with an equal volume of normal saline into the left ear margin vein as its own negative control, administered once a day for 3 consecutive days. 24 hours after the last administration, the rabbit hair at the administration site and surrounding tissues was removed, and after alcohol disinfection, the blood vessels and surrounding tissues were cut off, rinsed with normal saline, fixed in formalin for 24 hours, and then examined for pathological sections. The atlases of the rabbit's ear veins in the control group, raw material drug group and preparation group are shown in Figure 2, Figure 3 and Figure 4, respectively. It can be seen from Figure 2 that the vascular endothelial cells are degenerated, necrotic, and shedding, the blood vessels are clear, there is no thrombus in the tube, no inflammation, no bleeding and other lesions in the perivascular tissue; Figure 4 shows that there is a large amount of inflammatory cell infiltration in the perivascular tissue, but there is no large amount of cellulose exudation. It can be seen that the vascular irritation of bufalin raw material drug is relatively serious, while the vascular irritation of the preparation group is relatively weak, indicating that the n-butyl cyanoacrylate nanocarrier can enhance the safety of medication, and this conclusion is also consistent with that obtained in Experimental Example 3. The conclusion is the same.

The described embodiment is a preferred implementation of the present invention, but the present invention is not limited to the above-mentioned implementation, without departing from the essence of the present invention, any obvious improvement, replacement, replacement, etc. that those skilled in the art can make Modification and transformation all belong to the protection scope of the present invention.

Claims (10)

1. a medicine carrying polyalkylcyanoacrylate nano-carrier, is characterized in that, by mass percentage calculate comprise following component: the medicine of 1 ~ 10%, the stabilizing agent of 25 ~ 75%, 9 ~ 50% alpha-cyanoacrylate alkane ester monomer and 9 ~ 30% freeze drying protectants.

2. a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 1, is characterized in that,

Described medicine is: in Toadpoison Medicine, mitoxantrone, curcumin, paclitaxel, etoposide, paclitaxel of dwelling, gefitinib, hydroxy camptothecin, amycin, fluorouracil, cyclophosphamide, irinotecan or cisplatin more any one.

3. a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 1, is characterized in that,

Described stabilizing agent is: PLURONICS F87, poloxamer188, pluronic F127, macrodex, Dextran 40, Dextran-20, polyvinyl alcohol series, tween 80, any one or mixture several arbitrarily in Arlacel-80.

4. a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 1, is characterized in that,

Described alpha-cyanoacrylate alkane ester is: any one in methyl 2-cyanoacrylate, ethyl ester, butyl ester, isobutyl ester, own ester or dissident's ester.

5. a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 1, is characterized in that, the mean diameter of described medicine carrying polyalkylcyanoacrylate nano-carrier is 20 ~ 1000nm.

6. a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 5, is characterized in that, the mean diameter of described medicine carrying polyalkylcyanoacrylate nano-carrier is 50 ~ 200nm.

7. the preparation method of a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 1, is characterized in that, comprise the following steps:

(1) medicine is placed in dense alcohol system together with alpha-cyanoacrylate alkane ester monomer to be polymerized:

Calculate by mass percentage, take 1 ~ 10% medicine, be dissolved in 5mL to contain 4.5 ~ 45% stabilizing agents or do not contain in the dense alcohol system of stabilizing agent, i.e. ethanol-water system, wherein concentration of alcohol is 89 ~ 99%, and control rotating speed is 100 ~ 1000rpm, slowly joins in described dense alcohol system by 9 ~ 50% alpha-cyanoacrylate alkane ester monomers dropwise, stirring at room temperature 5h, obtains preliminary polymerization objects system in alcohol;

(2) preparation of aqueous phase:

Calculate by mass percentage, take 4 ~ 50% stabilizing agent distilled water and dissolve, or do not add stabilizing agent, be adjusted to specific pH2 ~ pH7.4, this is aqueous phase;

(3) described preliminary polymerization objects system is slowly dropwise transferred in described aqueous phase:

Controlled to be 100 ~ 1000rpm by the rotating speed of described aqueous phase, dropwise joined in aqueous phase by described preliminary polymerization objects system, adopt water bath sonicator that system is uniformly dispersed, wherein water bath sonicator temperature is 0 ~ 50 DEG C, and the power of water bath sonicator is 10 ~ 200W;

(4) rotary evaporation:

After described step (3) gained system is continued stirring 0.5 ~ 2h, regulate pH to neutral, rotary evaporation in vacuo removing ethanol, the temperature of rotary evaporation is 0 ~ 60 DEG C;

(5) probe sonication:

Adopt probe sonication, shear the nanoparticle in described step (4) gained system, to obtain the little nanoparticle of size, wherein the power of probe sonication is 10 ~ 1000W, and probe sonication number of times is 10 ~ 600 times;

(6) filter:

Be 0.8 μm of membrane filtration by aperture, obtain medicine carrying polyalkylcyanoacrylate nanoparticle suspension;

(7) lyophilization:

In described medicine carrying polyalkylcyanoacrylate nanoparticle suspension, add 9 ~ 30% freeze drying protectants, lyophilizing obtains powder in 24 hours.

8. the preparation method of a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 7, it is characterized in that, described water bath sonicator temperature is 10 ~ 30 DEG C, and described water bath sonicator power is 80 ~ 100W.

9. the preparation method of a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 7, it is characterized in that, described rotating evaporation temperature is 10 ~ 30 DEG C.

10. the preparation method of a kind of medicine carrying polyalkylcyanoacrylate nano-carrier according to claim 7, it is characterized in that, described probe sonication power is 100 ~ 400W, and described probe sonication number of times is 50 ~ 400.