WO2018188136A1

WO2018188136A1 - Method for preparing ultra-high drug-loaded nanoparticles by means of sequence precipitation-complex coacervation

Info

Publication number: WO2018188136A1
Application number: PCT/CN2017/083210
Authority: WO
Inventors: 刘东飞; 桑托斯•海尔德•A.; 凡进; 殷国勇
Original assignee: 刘东飞; 桑托斯•海尔德•A.; 凡进; 殷国勇
Priority date: 2017-04-12
Filing date: 2017-05-05
Publication date: 2018-10-18
Also published as: CN108685872B; CN108685872A

Abstract

A method for preparing ultra-high drug-loaded nanoparticles by means of sequence precipitation-complex coacervation. The method comprises: first, mix a first reactant and a second reactant in a first reaction vessel (1), and then precipitate active pharmaceutical ingredients in the first reaction vessel (1) to form cores of drug nanoparticles; the reaction product in the first reaction vessel (1) quickly flows into a second reaction vessel (2) to mix with a third reactant; and after undergoing a complex coacervation reaction, shell materials are deposited on the surfaces of the cores of drug nanoparticles so as to form ultra-high drug-loaded nanoparticles having a core-shell structure.

Description

Method for preparing ultrahigh drug-loaded nanoparticles by sequence precipitation complexation coacervation method

Technical field

The invention relates to a method for preparing ultra-high drug-loaded nanoparticles by a sequence precipitation complexation coacervation method, and belongs to the technical field of pharmaceutical preparations.

Background technique

The development of nanotechnology has greatly accelerated the research of medical science. The application of nanotechnology to the prevention, diagnosis and treatment of diseases is called nanomedicine. The global market for nanomedicine will increase from $5.5 billion in 2011 to $12.7 billion in 2016. Nano drug delivery systems, such as nanoparticle carriers, consist primarily of lipid materials and/or polymers, and the therapeutic agents they encase ^[1] , which can improve the efficacy of traditional drugs. Although some nano drug delivery systems have been successfully applied to the diagnosis and treatment of clinical diseases, such as cancer, pain and infection, there are still many aspects to be developed in the development of nano drug delivery systems. The clinical transformation of nano-delivery systems still faces many extremely difficult problems. The most important of these is the precise control of the physicochemical properties of the prepared nano-delivery system, the reproducibility between batches and batches, and the feasibility of industrial scale-up. Another major challenge is that most nano drug delivery systems consist primarily of non-therapeutic carrier materials, so the delivery of clinically effective therapeutic doses of drugs requires large amounts of nanocarriers. At the same time, the carrier material itself is at risk of causing side effects and greatly increases the cost of disease treatment. Therefore, there is an urgent need to develop an advanced method for precisely controlling the preparation process of nanoparticles, making the prepared nanoparticles have a very narrow particle size distribution, an ultra-high drug loading amount, a controlled drug release, and an ultra-high yield.

Due to the small pipe diameter and the large surface area to volume ratio brought about by it, the microfluidic device can achieve rapid and uniform mass transfer, which can precisely control the physicochemical properties of the prepared nanoparticles. For the precise control of the preparation process and the synthesis of continuous nanoparticles, the microfluidic technology can prepare nanoparticles with narrow particle size distribution, the difference between batches of nanoparticles is low, and the preparation method is convenient for industrial amplification. Although the preparation of nanoparticles by microfluidic technology has the above advantages, the drug loading of the prepared nanoparticles is still at a relatively low level. Taking polylactic acid-glycolic acid copolymer (PLGA) nanoparticles prepared by microfluidic technology as an example, the drug loading of paclitaxel is only about 5% ^[2] , and the drug loading of docetaxel is from ~1% to ~7%. Not waiting ^[3] . Therefore, the development of microfluidic nanoparticle preparation methods with ultra-high drug loading is very important for nanomedicine.

The theoretical drug loading of drug nanocrystals (the size of drug crystals at the nanometer level) is 100% ^[4] . Due to the very small particle size and the large specific surface area, the drug nanocrystalline particles have the characteristics of rapid dissolution and drug release. Although drug nanocrystalline particles have been widely studied and applied to the treatment of clinical diseases for more than 30 years, the controlled release of nanocrystalline drugs has not been achieved ^[5] . In addition, the drug nanocrystalline particles are highly dispersed multiphase systems with a large specific surface area, and a stabilizer is required to maintain the colloidal stability of the drug nanocrystalline particles. The stabilizer adsorbs on the surface of the drug nanocrystal particles, and utilizes steric hindrance to avoid aggregation of the drug nanocrystal particles. Stabilizers for the preparation of nanomedicine crystal particles mainly include surfactants and hydrophilic polymer materials. Common non-particle stabilizers are mainly poloxamer, Tween, Span, hydroxypropylmethylcellulose (HPMC). ), hydroxypropyl cellulose (HPC), polyvinyl alcohol (PVA), etc.; ionic stabilizers are sodium dodecyl sulfate (SDS), phospholipids and cholic acid derivatives ^[4] . The combination of two or more stabilizers can produce synergistic effects, but some specific stabilizers will cause aggregation of drug nanocrystalline particles due to interaction ^[6] . The choice of stabilizers and their combinations and concentrations has led to a large number of prescription optimization efforts. More importantly, for each particular drug nanocrystalline particle, the choice of stabilizer is specific, that is, optimization of the stabilizer is required separately.

Summary of the invention

The technical problem to be solved by the invention is to provide a method for preparing ultra-high drug-loaded nanoparticles by ultra-fast sequence precipitation complex coacervation method, which can well solve some problems faced by traditional nano drug delivery systems.

In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

The first sequence of precipitation complexation coacervation method for preparing ultrahigh drug-loaded nanoparticles, first mixing the first reactant and the second reactant in the first reactor, so that the active pharmaceutical ingredient is rapidly in the first reactor Precipitating to form a drug nanoparticle core; the reaction product of the first reactor rapidly flows into the second reactor, is mixed with the third reactant, and the shell material is complexed and agglomerated and deposited on the surface of the core of the drug nanoparticle to form Ultra-high drug-loaded nanoparticles with core-shell structure;

among them,

The first reactant is solvent I, and the solvent I is a poor solvent of the active pharmaceutical component and is a benign solvent of the first outer shell material;

The second reactant is a solution formed by the active pharmaceutical ingredient, the first outer shell material and the solvent II, that is, the solvent II is an active drug. a benign solvent of the composition and the first outer shell material;

The third reactant is a solution formed by the second outer shell material and the solvent III, and the solvent III is a poor solvent of the active pharmaceutical component and is a benign solvent of the first outer shell material and the second outer shell material;

The solvents I, II and III are mutually soluble;

The first outer shell material and the second outer shell material can undergo a complex condensation reaction;

In the first reactor, the inlet flow rate of the first reactant is greater than the inlet flow rate of the second reactant to ensure that the volume ratio of the solvent I to the solvent II is greater than 1:1, thereby ensuring the first reactant and During the mixing of the second reactant, the active drug can be completely precipitated to form the active drug nanoparticle core.

Wherein the ultra-high drug-loaded nanoparticle comprises a nano-core containing an active pharmaceutical ingredient and an outer shell for controlling drug release; wherein the mass of the active pharmaceutical ingredient accounts for 30-85%, preferably 35-75, of the mass of the entire nanoparticle. %, most preferably 45-70%; said ultrahigh drug-loaded nanoparticles have a particle size of from 50 to 2000 nm, preferably from 80 to 800 nm, most preferably from 80 to 400 nm.

Wherein, the active pharmaceutical ingredient is a poorly water-soluble drug. The poorly water-soluble drugs include, but are not limited to, paclitaxel, docetaxel, doxorubicin, vincristine, camptothecin, hydroxycamptothecin, etoposide, curcumin, retinoic acid, fluorouracil, A Aminopterin, teniposide, daunorubicin, aclarithromycin, sorafenib, methylprednisone, minocycline, dexamethasone, cisplatin, atorvastatin, simvastatin , lovastatin, amiodarone, carbamazepine, carvedilol, chlorpromazine, cisapride, dapsone, azithromycin, neomycin, amphotericin B, griseofulvin, celecoxib , raloxifene, flurbiprofen, indomethacin, ibuprofen, tamoxifen, diclofenac, naproxen, piroxicam, ralivamide, efavirenz, nelfinavir, ar Zanavir, ritonavir, sirolimus, spironolactone, tacrolimus, talirolol, terfenadine, estradiol, vitamin A, vitamin D, vitamin E, vitamin K, Any one or a mixture of cyclosporine or insulin. Preferred paclitaxel, docetaxel, vincristine, camptothecin, hydroxycamptothecin, curcumin, retinoic acid, sorafenib, methylprednisone, minocycline, atorvastatin, simvastatin Statins, lovastatin, amphotericin B, griseofulvin, celecoxib, indomethacin, ibuprofen, diclofenac, naproxen, piroxicam, vitamin A, vitamin D, vitamin E, vitamin K , cyclosporine or insulin; most preferred are paclitaxel, curcumin, sorafenib, celecoxib or cyclosporine.

Wherein, the outer shell for controlling drug release includes, but not limited to, a polymer or a protein or a mixture of any one or more of deoxyribonucleic acid or ribonucleic acid. Wherein, the polymer includes, but is not limited to, chitosan and its derivatives, hypromellose acetate succinate, methylcellulose, hyaluronic acid, heparin, poly-2-acrylamide-2-methyl Propane, kondagogu gum, pectin, xanthan gum, poly D-glutamic acid, dextran sulfate, carrageenan, carboxymethyl cellulose, sodium carboxymethyl cellulose, polyethyleneimine and its derivatives, fine Amines and derivatives thereof, polylysine and its derivatives, polyaminos and derivatives thereof, cationic cyclodextrin and its derivatives, cationic phospholipids, cationic dextran and its derivatives, polystyrene sulfonate, poly Diallyldimethyl, N-methylated polyvinylpyridine, polyvinyl alcohol sulfuric acid, polydiallyldimethylammonium chloride, Eudragit E, carrageenan, polymethacrylic acid, poly N Any of -ethyl-4-vinylpyridine bromide, alginate, pectate, dextran, sulfated cellulose, guar gum, glutamic acid dextran, gum arabic or propylene glycol alginate a mixture of one or several. Preferred are chitosan and its derivatives, hyaluronic acid, poly-D-glutamic acid, polyethyleneimine and its derivatives, cationic cyclodextrin and its derivatives, cationic phospholipids, cationic dextran and its derivatives or alginic acid Salt; most preferred are chitosan and its derivatives, polyethyleneimine and its derivatives or cationic dextran and its derivatives. Wherein, the protein includes, but is not limited to, any one or a few of albumin, collagen, gelatin, elastin, gliadin, legumin, zein, soy protein, milk protein, whey protein Kind of mixture. Albumin, collagen, gelatin, milk protein and whey protein are preferred; albumin is most preferred. Wherein, the deoxyribonucleic acid or ribonucleic acid includes, but is not limited to, small interfering RNA, small hairpin RNA or plasmid DNA. Most preferred is plasmid DNA.

The preferred way is:

The first outer shell material is chitosan, and the second outer shell material is sodium alginate, hyaluronic acid, heparin, poly 2-acrylamide-2-methylpropane, kondagogu gum, pectin, yellow Raw gum, poly-D-glutamic acid, dextran sulfate, carrageenan, carboxymethyl cellulose or sodium carboxymethyl cellulose;

Alternatively, the first outer shell material is polyethyleneimine and its derivatives, spermine and its derivatives, polylysine and its derivatives, polyurethane and its derivatives, cationic cyclodextrin and its derivatives. Or a cationic phospholipid, or a cationic dextran and a derivative thereof, wherein the second outer shell material is deoxyribonucleic acid or ribonucleic acid;

Alternatively, the first outer shell material is polystyrene sulfonate, and the second outer shell material is polydiallyl dimethyl;

Or the first outer shell material is carboxymethyl cellulose, and the second outer shell material is N-methylated polyvinyl pyridine;

Alternatively, the first outer casing material is polyvinyl alcohol sulfuric acid, and the second outer casing material is polydiallyldimethylammonium chloride;

Alternatively, the first outer casing material is Eudragit E, and the second outer casing material is carrageenan;

Or the first outer shell material is polymethacrylic acid, and the second outer shell material is poly N-ethyl-4-vinylpyridine bromide;

Alternatively, the first outer shell material is albumin, and the second outer shell material is dextran sulfate, sodium carboxymethyl cellulose, alginate, pectate, pectin, cationic polyglutamine. Acid (PGlu) or dextran;

Alternatively, the first outer shell material is β-lactoglobulin, and the second outer shell material is sodium carboxymethyl cellulose, sulfated cellulose, carrageenan, guar gum, glutamic acid Sugar, gum arabic, dextran sulfate or propylene glycol alginate.

The solvent I is water, or a buffer solution, or an aqueous solution containing an organic solvent; the solvent II is an organic solvent; and the solvent III is water or a buffer.

Wherein, the buffer includes, but not limited to, a hydrochloride buffer, a borate buffer, a nitrate buffer, a sulfate buffer, a phosphate buffer, a citrate buffer, a carbonate buffer, Acetate buffer, barbiturate buffer, Tris (tris) buffer, 2-(N-morpholine) ethanesulfonic acid buffer, hydroxyethylpiperazine ethanesulfonic acid buffer , ammonium chloride buffer, ethylenediamine buffer or triethylamine buffer. Preferred is hydrochloride buffer, phosphate buffer, citrate buffer, carbonate buffer, acetate buffer, MES buffer, HEPES buffer or triethylamine buffer, most preferably hydrochloride buffer Liquid, phosphate buffer, acetate buffer or triethylamine buffer.

Wherein, the organic solvent includes, but not limited to, methanol, ethanol, ethylene glycol, diethylene glycol, isopropanol, 1-propanol, 1,2-propanediol, 1,3-propanediol, butanol, 1, 2-butanediol, 1,3-butanediol, 1,4-butanediol, 1,5-pentanediol, 2-butoxyethanol, glycerin, methyldiethanolamine, diethanolamine, acetone, acetonitrile , diethylenetriamine, dimethoxyethane, ethylamine, dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, acetaldehyde, pyridine, triethylene glycol, ethyl acetate, dimethyl carbonate, dichloro a mixture of any one or more of methane, cyclohexane, n-octanol or chloroform. Preference is given to methanol, ethanol, ethylene glycol, isopropanol, 1,2-propanediol, acetone, acetonitrile, dimethyl sulfoxide or tetrahydrofuran, most preferably ethanol, acetonitrile, acetone or dimethyl sulfoxide.

The concentration of the organic solvent in the aqueous solution containing the organic solvent is in the range of 10 to 40% (v/v). Preference is given to 10% methanol solution, 20% methanol solution, 10% ethanol solution, 20% ethanol solution, 10% acetone solution or 20% acetone solution, most preferably 20% ethanol solution or 20% acetone solution.

Wherein, the first reactor has a residence time of 0.01 s to 500 s, preferably 0.05 to 100 s, and most preferably 0.1 to 10 s. The residence time of the second reactor is not required by the present invention.

Among them, the Reynolds number of the entire reaction system is from 1 to 2,000, preferably from 10 to 1,500, and most preferably from 10 to 1300. Reynolds number (Re) A dimensionless number that can be used to characterize fluid flow. Re=ρvL/μ, ρ, μ are fluid density (kg/m ³ ) and kinematic viscosity coefficient (N·s/m ² ), respectively, v and L are the characteristic velocity (m/s) and characteristic length of the flow field, respectively. (m). The Reynolds number physically represents the ratio of inertial force to viscous force level. For the mixing of the two fluids, the average flow velocity in the channel is taken as the characteristic velocity of the flow field.

The three reactants may flow into / s range of 1.0E-04m ³ regulate the 1.0E-09m ³ / s. High flow rates result in different mixing modes, from microvortex to turbulent jets. Especially after the fluid reaches the turbulent jet, the different fluids will reach complete mixing in a very short time. Rapid, thorough mixing allows the fluid to be mixed for a shorter period of time than the active drug and the shell material. Thus, precipitation of the active drug as well as the outer shell material can be carried out in a homogeneous liquid mixture to form an active drug nanoparticle core having a relatively uniform particle size distribution, as well as core-shell nanoparticles. At low flow rates, the core-shell nanoparticles formed have larger particle sizes and wider particle size distribution. Under high flow conditions, the core-shell structure has smaller particle size and particle size. The distribution is narrow.

The second sequence precipitation complexation coacervation method for preparing ultra-high drug-loaded nanoparticles, first mixing the first reactant and the second reactant in the first reactor, so that the active pharmaceutical ingredient is rapidly in the first reactor The precipitate forms a core of the drug nanoparticle; the reaction product of the first reactor rapidly flows into the second reactor, is mixed with the third reactant, and the shell material is subjected to a sequence precipitation complex coacervation reaction and deposited on the surface of the drug nanoparticle core. Forming ultra-high drug-loaded nanoparticles having a core-shell structure;

among them,

The first reactant is a solution formed by both the first outer shell material and the solvent IV, and the solvent IV is a poor solvent of the active pharmaceutical ingredient and is a benign solvent of the first outer shell material;

The second reactant is a solution formed by both the active pharmaceutical ingredient and the solvent V;

The third reactant is a solution formed by the second outer shell material and the solvent VI, wherein the solvent VI is a poor solvent of the active pharmaceutical component and is a benign solvent of the first outer shell material and the second outer shell material;

The solvents IV, V and VI are mutually soluble;

In the first reactor, the inlet flow rate of the first reactant is greater than the inlet flow rate of the second reactant to ensure that the volume ratio of the solvent IV to the solvent V is greater than 1:1, thereby ensuring the first reactant and During the mixing of the second reactant, the active drug can be completely precipitated to form the active drug nanoparticle core.

Preferably,

Wherein, the solvent IV is water, or a buffer solution, or an aqueous solution containing an organic solvent; the solvent V is an organic solvent; and the solvent VI is water or a buffer.

Among them, the Reynolds number of the entire reaction system is from 1 to 2,000, preferably from 10 to 1,500, and most preferably from 10 to 1300.

In the second preparation method, the definition and usage rules of the Reynolds number are the same as the first preparation method.

The three reactants may flow into / s range of 1.0E-04m ³ regulate the 1.0E-09m ³ / s. The high flow rate brings different mixing modes, from laminar flow to turbulent jets. Especially after the fluid reaches the turbulent jet, the different fluids will reach complete mixing in a very short time. Rapid, thorough mixing allows the fluid to be mixed for a shorter period of time than the active drug and the shell material. Thus, precipitation of the active drug as well as the outer shell material can be carried out in a homogeneous liquid mixture to form an active drug nanoparticle core having a relatively uniform particle size distribution, as well as core-shell nanoparticles. At low flow rates, the core-shell nanoparticles formed have larger particle sizes and wider particle distribution. Under high flow conditions, the core-shell structure has smaller particle size and particle distribution. Narrower.

In the preparation method of the present invention, the first and second outer shell materials are water-soluble substances, which avoids the use of an organic solvent, so that the poorly soluble active pharmaceutical ingredient can be more completely precipitated in the first reactor, and also makes it difficult. The choice of soluble active pharmaceutical ingredients is more extensive.

In the preparation method of the present invention, when the outer shell material is deoxyribonucleic acid or ribonucleic acid, the combined administration of small molecule chemical drugs and nucleic acid drugs can be realized, and the synergistic therapeutic effect of the disease can be achieved, and the dosage and the drug are reduced. Toxic side effects for a more effective treatment.

The invention also discloses an ultra-high drug-loading nanoparticle preparation device, comprising a first reactor and a second reactor; the first reactor comprises a first inlet, a second inlet and a first outlet, and the second reactor comprises a first reactor Three imports and fourth imports;

The first inlet is for adding a first reactant or a second reactant;

The second inlet is for adding a second reactant or a first reactant;

The first reactor is configured to accommodate a reaction between the first reactant and the second reactant;

a first outlet of the first reactor is in communication with a fourth inlet of the second reactor for conveying a reaction product of the first reactant and the second reactant into the second reactor;

The third inlet is for adding a third reactant;

The second reactor is for containing a third reactant to react with a reaction product of the first reactant and the second reactant.

Wherein the first inlet of the first reactor communicates with the pre-receiving chamber for accommodating the first reactant.

Wherein, the first reactor and/or the second reactor are internally provided with a herringbone for acceleration.

Wherein the first reactor and/or the second reactor are arranged in a zigzag bent structure.

Wherein the first reactor and/or the second reactor are arranged in a hairpin-like curved structure.

The poorly water-soluble drug according to the present invention refers to a drug in which 1 g of a drug needs to be completely dissolved in water of 1000 ml or more.

The benign solvent of the present invention means that 1 g of the solute can be completely dissolved in a solvent of 100 ml or less.

The poor solvent according to the present invention means that 1 g of the solute is required to be completely dissolved in a solvent of 1000 ml or more.

Advantageous Effects: The present invention has the following advantages over the prior art:

1. The phenomenon of agglomeration and sedimentation occurs in the drug nanoparticles prepared by the prior art. It is necessary to disperse the nanometer-sized drug particles in water by the stabilizing action of the surfactant or the polymer material, thereby forming a relatively stable colloidal dispersion system. The selection of the type and concentration of surfactants in the prior art preparation process has led to a large number of prescription optimization work. The invention has the advantages that after the pharmaceutically active substance nanoparticles are formed in the first reactor, they quickly enter the second reactor, and form an outer layer of the polymer shell on the surface of the drug nanoparticles in a very short time to stabilize the drug. Nanoparticles. The invention makes the preparation of the drug nanometers without using a stabilizer, which greatly reduces the workload of prescription optimization.

2. Preparation of core-shell nanoparticles usually requires two steps: preparing drug nanoparticles and encapsulating the drug nanoparticles in the outer core shell. In the prior art, these two steps usually require a lot of intermediate processes, such as centrifugation, concentration, sonication, vortexing, and long-term incubation of the prepared drug nanoparticles, which will give the final prepared core. The shell-structured nanoparticles bring a lot of uncertainty. Since each step makes the formed nanoparticles have a certain particle size distribution, the particle size distribution of the core-shell structured nanoparticles prepared by the conventional method is greatly increased as compared with the rapid continuous sequence precipitation method of the present invention. The rapid continuous sequence precipitation method of the invention can produce nano-shells of core-shell structure in one time, and greatly enhances the control of the physical and chemical properties of the finally prepared nanoparticles.

3. The drug-loaded nanoparticles prepared by the existing microfluidic method or the emulsified volatilization method can release the drug well. However, since the drug and the polymer are simultaneously precipitated, the space in which the pharmaceutically active substance can be encapsulated in the polymer matrix is limited, and a high drug loading amount cannot be obtained. The invention enables the drug nanoparticles to be formed first, and then forms a polymer shell on the surface thereof, so that the drug loading amount of the drug is greatly improved, and at the same time, the drug release effect is controlled.

4. The first and second outer shell materials in the preparation method of the present invention are all water-soluble substances, avoiding the use of an organic solvent, so that the poorly soluble active pharmaceutical ingredient can be more completely precipitated in the first reactor, and at the same time It also makes the selection of poorly soluble active pharmaceutical ingredients more extensive.

5. In the preparation method of the present invention, when the outer shell material is deoxyribonucleic acid or ribonucleic acid, the combined administration of small molecule chemical drugs and nucleic acid drugs can be achieved, and the synergistic therapeutic effect of the disease can be achieved, and the dosage and the dosage are reduced. The toxic side effects of the drug achieve a more effective therapeutic effect.

DRAWINGS

1 is a schematic view of an apparatus for preparing an ultra-high drug-loaded nanoparticle of Example 1. The reactor of Example 1 was made of glass, polycarbonate or polytetrafluoroethylene.

2 is a schematic view of an apparatus for preparing an ultra-high drug-loaded nanoparticle of Example 2. The reactor material of Example 2 was a polymer material such as polydimethylsiloxane (PDMS) or polycarbonate.

3 is a schematic view of an apparatus for preparing an ultra-high drug-loaded nanoparticle of Example 3. The reactor of Example 3 was made of a polymer material such as PDMS or polycarbonate.

4 is a schematic view of an apparatus for preparing an ultra-high drug-loaded nanoparticle of Example 4. The reactor material of Example 4 was made of a polymer material such as PDMS or polycarbonate.

5 is a schematic view of an apparatus for preparing an ultra-high drug-loaded nanoparticle of Example 5. The reactor of Example 5 was made of a polymer material such as PDMS or polycarbonate.

6 is a schematic view of an apparatus for preparing an ultra-high drug-loaded nanoparticle of Example 6. The reactor of Example 6 was made of a polymer material such as PDMS or polycarbonate.

7 is a schematic view of an apparatus for preparing an ultra-high drug-loaded nanoparticle of Example 7. The reactor material of Example 7 was made of a polymer material such as PDMS or polycarbonate.

Figure 8. Cationic phospholipids and plasmid DNA complexes (PPDNA) encapsulated paclitaxel (PTX) nanocrystalline nanoparticles (PTX@PPDNA) Drug release profile, temperature is 37 ° C, (n = 3)

Figure 9. Drug release profile of curcumin (CUR) nanocrystal nanoparticles (CUR@PAD-DNA) encapsulated with cationic dextran (PAD) and plasmid DNA complex at 37 ° C (n = 3).

Figure 10. Drug release profile of polyethylenimine (PEI) and DNA plasmid complexes coated with sorafenib (SFN) nanocrystal nanoparticles (SFN@PEI-DNA) at 37 ° C (n = 3).

Figure 11 shows the drug release profile of celecoxib (CEL) nanocrystalline nanoparticles (CEL@BSAPGlu) condensed with albumin (BSA) and cationic polyglutamic acid (PGlu) at a temperature of 37 ° C, (n = 3).

Figure 12 shows the drug release profile of cationic dextran (PAD) and plasmid DNA complexes coated with cyclosporin A (CSA) nanocrystal nanoparticles (CSA@PAD-DNA) at 37 ° C, (n = 3).

detailed description

The invention can be better understood in light of the following examples. However, those skilled in the art will understand that the description of the embodiments is only intended to illustrate the invention and should not be construed as limiting the invention as described in the claims.

Example 1

As shown in FIG. 1 , the embodiment discloses an ultra-high drug-loading nanoparticle preparation device, which comprises a first reactor 1 and a second reactor 2; the first reactor includes a first inlet 1a, a second inlet 1b, and The first outlet 1c, the second reactor includes a third inlet 2a and a fourth inlet 2b.

Option 1 case: the first inlet 1a is used to add a first reactant, the first reactant enters the first reactor 1 through the first inlet 1a; the second inlet 1b is used to add a second reactant, The second reactant enters the first reactor 1 in the direction of the arrow next to the second inlet 1b in Fig. 1; the first reactor 1 is used to accommodate the reaction of the first reactant and the second reactant. Option 2: the first inlet 1a is for adding a second reactant, the second reactant is introduced into the first reactor 1 through the first inlet 1a; the second inlet 1b is for adding a first reactant, A reactant enters the first reactor 1 in the direction of the arrow next to the second inlet 1b in Fig. 1; the first reactor 1 is adapted to accommodate the reaction of the first reactant and the second reactant.

The first outlet 1c of the first reactor 1 is in communication with the fourth inlet 2b of the second reactor for conveying the reaction product of the first reactant and the second reactant into the second reactor 2, FIG. The first outlet 1c coincides with the position of the fourth inlet 2b; the third inlet 2a is used to add a third reactant, and the third reactant enters the second reactor 2 in the direction of the arrow next to the second inlet 2a in FIG. The second reactor 2 is configured to contain a third reactant to react with a reaction product of the first reactant and the second reactant. This example is directed to a reactor prepared using glass, polycarbonate or polytetrafluoroethylene.

The first inlet of the first reactor communicates with the pre-receiving chamber 3 for containing the first reactant.

Example 2

As shown in FIG. 2, the difference between this embodiment and the embodiment 1 is that the front accommodating chamber 3 is not used, and the first inlet and the first reactor are disposed on the same straight line, and the first reactor and the second reactor are disposed at On the same line. The second inlet is two, perpendicular to the straight line formed by the first inlet and the first reactor. The third inlet is two, perpendicular to the line formed by the first reactor and the second reactor, respectively. The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet, respectively, or the first reactant and the second reactant enter the first reactor through the second inlet and the first inlet, respectively. The exchange of the tubes in which the first reactant and the second reactant enter the first reactor does not affect the preparation of the nanoparticles. This embodiment is directed to a reactor prepared using a polymer material such as PDMS. The middle dotted line indicates that the length of the reactor can be adjusted.

Example 3

As shown in FIG. 3, the difference between this embodiment and the second embodiment is that the second inlet is one, and the first inlet and the second inlet are respectively obliquely connected to the first reactor, forming an incident with the first reactor. The angle is between 0 and 180 degrees, and the rest is the same. The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet, respectively, or the first reactant and the second reactant enter the first reactor through the second inlet and the first inlet, respectively. The exchange of the tubes in which the first reactant and the second reactant enter the first reactor does not affect the preparation of the nanoparticles. This embodiment is directed to a reactor prepared using a polymer material such as PDMS. The middle dotted line indicates that the length of the reactor can be adjusted.

Example 4

As shown in Fig. 4, this embodiment differs from the embodiment 3 in that the first reactor and/or the second reactor are internally provided with a herringbone for acceleration, and the remaining portions are identical. The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet, respectively, or the first reactant and the second reactant enter the first reactor through the second inlet and the first inlet, respectively. Passing the first reactant and the second reactant Pipeline exchange into the first reactor does not affect the preparation of the nanoparticles. This embodiment is directed to a reactor prepared using a polymer material such as PDMS. The role of the herringbone is to accelerate the mixing speed of the reactants, especially for the preparation of nanoparticles under the condition that the Reynolds number is less than 200, and the nanoparticles with relatively uniform particle distribution can be obtained. When the Reynolds number is higher than 500, even if there is no herringbone, the speed of mixing is very fast. Of course, the amount of herringbone in this device can be more.

Example 5

As shown in FIG. 5, the difference between this embodiment and Embodiment 2 is that both the second inlet and the third inlet are one, and the first inlet and the second inlet are obliquely connected to the first reactor, respectively, and react with the first reaction. The devices all form an incident angle (0 to 180 degrees). The first reactor and/or the second reactor are arranged in a zigzag bent structure. The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet, respectively, or the first reactant and the second reactant enter the first reactor through the second inlet and the first inlet, respectively. The exchange of the tubes in which the first reactant and the second reactant enter the first reactor does not affect the preparation of the nanoparticles. This embodiment is directed to a reactor prepared using a polymer material such as PDMS. The role of the zigzag pipeline is to accelerate the mixing speed of the reactants, especially for the preparation of nanoparticles under the condition that the Reynolds number is less than 200, and the nanoparticles with relatively uniform particle distribution can be obtained. When the Reynolds number is greater than 500, the mixing speed is very fast even without the zigzag pipe. Similarly, the length of the zigzag pipe in this device can be more.

Example 6

As shown in FIG. 6, this embodiment differs from Embodiment 1 in that the front housing chamber 3 is not employed. The second inlet is two, perpendicular to the straight line formed by the first inlet and the first reactor. The third inlet is two, perpendicular to the line formed by the first reactor and the second reactor, respectively. The first reactor and/or the second reactor are arranged in a hairpin-like curved structure. The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet, respectively, or the first reactant and the second reactant enter the first reactor through the second inlet and the first inlet, respectively. The exchange of the tubes in which the first reactant and the second reactant enter the first reactor does not affect the preparation of the nanoparticles. This embodiment is directed to a reactor prepared using a polymer material such as PDMS. The dotted line in the middle indicates that the number of repetitions of the card issuance structure can be more. The effect of the hairpin structure is the same as that of the herringbone groove, accelerating the mixing between the reactants.

Example 7

As shown in FIG. 7, the difference between the embodiment and the embodiment 6 is that the second inlet is one, and the first inlet and the second inlet are obliquely connected to the first reactor, respectively, and form an incident with the first reactor. Angle (0 to 180 degrees). The third inlet is one and is obliquely connected to the second reactor. The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet, respectively, or the first reactant and the second reactant enter the first reactor through the second inlet and the first inlet, respectively. The exchange of the tubes in which the first reactant and the second reactant enter the first reactor does not affect the preparation of the nanoparticles. This design is primarily for reactors prepared using polymeric materials such as PDMS. The dotted line in the middle indicates that the number of repetitions of the card issuance structure can be more. The effect of the hairpin structure is the same as that of the herringbone groove, accelerating the mixing between the reactants.

Example 8

The present invention can be used to prepare ultra-high drug-loaded core-shell nanoparticles by using any of the devices of Examples 1-7. The device may be made of germanium, silicon wafer, glass, quartz, PDMS, plexiglass (polymethyl methacrylate PMMA) or vinyl polymer, polycarbonate (PC), polytetrafluoroethylene, metal, ceramics, etc. production. The vinyl polymer may be selected from polystyrene PS, polyethylene PE, polyvinyl chloride PVC, polydichloroethylene PVDC, and the like. The conduit in the device can be composed of any material suitable for the fluid to flow therein. Typically, the pipe material is resistant to the solvent I-VI.

Example 9 (the first preparation method described in the present invention)

This example discloses the preparation of an ultra-high drug-loaded and controlled release drug nanoparticle, a cationic phospholipid and a plasmid DNA complex (PPDNA) coated with paclitaxel (PTX) nanocrystal nanoparticles (PTX@PPDNA).

PTX (5 mg/mL) and cationic phospholipid (1 mg/mL) in ethanol as the second reactant, aqueous solution (pH 7.0) and aqueous plasmid DNA (pH 7.0; nitrogen-phosphorus ratio, N/P=10) as the first And a third reactant. The flow ratio of the first, second and third reactants was 5:1:30 under a Reynolds number of 100. At this time, the average particle diameter of the nanoparticles was about 159.4 nm, and the drug loading was about 45.8%.

At the same time, the core-shell nanoparticles we prepared can control the drug release of the encapsulated drug nanoparticles. As shown in Fig. 8, the PTX nanocrystal itself has a rapid and complete release of the drug due to its large specific surface area; when the PTX nanocrystal is encapsulated by PPDNA, the release of the drug is greatly slowed down.

Example 10 (the first preparation method described in the present invention)

This example discloses the preparation of curcumin (CUR) nanocrystalline nanoparticles (CUR@PAD-DNA) coated with ultra-high drug-loaded and controlled release drug nanoparticles, cationic dextran (PAD) and plasmid DNA complexes. .

A solution of CUR (5 mg/mL) and PAD (1 mg/mL) in ethanol was used as the second reactant, and an aqueous solution of pH 7.0 and an aqueous solution of DNA (pH 7.0, N/P = 10) were used as the first and third reactants, respectively. The flow ratio of the first, second and third reactants was 5:1:30 under a Reynolds number of 100. At this time, the average particle diameter of the nanoparticles was about 169.2 nm, and the drug loading was 41.0%.

At the same time, the core-shell nanoparticles we prepared can control the drug release of the encapsulated drug nanoparticles. The shell polymer PAD we use is a pH sensitive polymer material. Under neutral conditions (pH 7.4), the outer shell structure remains intact and CUR is not released; and when the pH is lowered to acidic, CUR achieves a rapid release due to degradation of the outer shell PAD (Figure 9). Example 11 (the second preparation method described in the present invention)

This example discloses an ultra-high drug-loaded and controlled release drug nanoparticle, polyethyleneimine (PEI) and DNA plasmid complex coated with sorafenib (SFN) nanocrystalline nanoparticles (SFN@PEI-DNA) Preparation of).

A SFN (5 mg/mL) acetone solution was used as the second reactant, and a pH 7.0 buffer of PEI (1 mg/mL) and an aqueous plasmid DNA solution (pH 7.0; N/P = 10) was used as the first and third reactants, respectively. The flow ratio of the first, second and third reactants was 5:1:30 under a Reynolds number of 100. At this time, the average particle diameter of the nanoparticles was about 135.9 nm, and the drug loading was 53.9%.

At the same time, the core-shell nanoparticles we prepared can control the drug release of the encapsulated drug nanoparticles. As shown in FIG. 10, the SFN nanocrystal itself has a rapid complete release of the drug due to its large specific surface area; when the SFN nanocrystal is encapsulated by the PEI-DNA complex, the release of the drug is greatly slowed down.

Example 12 (the second preparation method described in the present invention)

This embodiment discloses an ultra-high drug-loaded nanoparticle, an aggregate of albumin (BSA) and cationic polyglutamic acid (PGlu), which encapsulates celecoxib (CEL) nanocrystalline nanoparticles (CEL@BSAPGlu). preparation.

A solution of CEL (5 mg/mL) in ethanol was used as the second reactant, and a BSA (1.0 mg/mL) solution (pH 7.0) and a PGlu solution (0.79 mg/mL, pH 7.0) were used as the first and third reactants, respectively. The flow ratio of the first, second and third reactants was 5:1:30 under a Reynolds number of 100. At this time, the average particle diameter of the nanoparticles was about 649.7 nm, and the drug loading was 38.2%.

At the same time, the core-shell nanoparticles we prepared can control the drug release of the encapsulated drug nanoparticles. As shown in Fig. 11, the SFN nanocrystal itself, due to its large specific surface area, rapidly released the drug; when the SFN nanocrystals were encapsulated by the PEI-DNA complex, the release of the drug was greatly slowed (Fig. 11).

Example 13 (the first preparation method described in the present invention)

This example discloses an ultra-high drug-loaded and controlled release drug nanoparticle, cationic dextran (PAD) and plasmid DNA complex coated with cyclosporin A (CSA) nanocrystalline nanoparticles (CSA@PAD-DNA) preparation.

A solution of CSA (4 mg/mL) and PAD (1 mg/mL) in ethanol was used as the second reactant, and an aqueous solution of pH 7.0 and an aqueous solution of DNA (pH 7.0, N/P = 10) were used as the first and third reactants, respectively. The flow ratio of the first, second and third reactants was 5:1:30 under a Reynolds number of 100. At this time, the average particle diameter of the nanoparticles was about 132.7 nm, and the drug loading was 44.7%.

At the same time, the core-shell nanoparticles we prepared can control the drug release of the encapsulated drug nanoparticles. The shell polymer PAD we use is a pH sensitive polymer material. Under neutral conditions (pH 7.4), the outer shell structure remained intact and CSA was not released; and when we lowered the pH to acidity, CSA reached a rapid release due to degradation of the outer shell PAD (Figure 12).

references

[1] T.M. Allen, P.R. Cullis, Science 2004, 303, 1818.

[2] D.F. Liu, S. Cito, Y. Z. Zhang, C. F. Wang, T. M. Sikanen, H. A. Santos, Adv. Mater. 2015, 27, 2298.

[3] a) N. Kolishetti, S. Dhar, PM Valencia, LQ Lin, R. Karnik, SJ Lippard, R. Langer, OCFarokhzad, Proc. Natl. Acad. Sci. USA 2010, 107, 17939; b) R. Karnik, F. Gu, P. Basto, C. Cannizzaro, L. Dean, W. Kyei-Manu, R. Langer, OCFarokhzad, Nano Lett. 2008, 8, 2906; c) JMLim, A. Swami , LMGilson, S. Chopra, S. Choi, J. Wu, R. Langer, R. Karnik, OCFarokhzad, ACS Nano 2014, 8, 6056.

[4] L. Gao, D.R. Zhang, M.H. Chen, J. Nanopart. Res. 2008, 10, 845.

[5] a) K. Fuhrmann, MAGauthier, JCLeroux, Mol. Pharmaceutics 2014, 11, 1762; b) L. Gao, GYLiu, JLMa, XQ Wang, L. Zhou, X. Li, J. Control .Release 2012,160,418.

[6] B. Van Eerdenbrugh, G. Van den Mooter, P. Augustijns, Int. J. Pharm. 2008, 364, 64.

Claims

A method for preparing ultrahigh drug-loaded nanoparticles by a sequence precipitation complexation coacervation method, characterized in that firstly, the first reactant and the second reactant are mixed in the first reactor, so that the active drug component is in the first reaction Precipitating to form a drug nanoparticle core; the reaction product of the first reactor rapidly flows into the second reactor, is mixed with the third reactant, and the shell material is complexed and agglomerated and deposited on the surface of the core of the drug nanoparticle. Forming ultra-high drug-loaded nanoparticles having a core-shell structure;

among them,

The first reactant is solvent I, and the solvent I is a poor solvent of the active pharmaceutical component and is a benign solvent of the first outer shell material;

The second reactant is a solution formed by the active pharmaceutical ingredient, the first outer shell material and the solvent II;

The third reactant is a solution formed by the second outer shell material and the solvent III, and the solvent III is a poor solvent of the active pharmaceutical component and is a benign solvent of the first outer shell material and the second outer shell material;

The solvents I, II and III are mutually soluble;

The first outer shell material and the second outer shell material can undergo a complex condensation reaction;

In the first reactor, the incoming flow rate of the first reactant is greater than the incoming flow rate of the second reactant.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 1, wherein the ultrahigh drug-loading nanoparticles comprise a nano-core containing an active pharmaceutical ingredient and control drug release The outer shell; wherein the mass of the active pharmaceutical ingredient accounts for 30-85% of the mass of the whole nanoparticle; and the ultrahigh drug-loaded nanoparticle has a particle diameter of 50-2000 nm.
The method according to claim 1, wherein the active pharmaceutical ingredient is a poorly water-soluble drug.
The method according to claim 3, wherein the poorly water-soluble drug comprises, but is not limited to, paclitaxel, docetaxel, doxorubicin, and the like. , vincristine, camptothecin, hydroxycamptothecin, etoposide, curcumin, retinoic acid, fluorouracil, methotrexate, teniposide, daunorubicin, aclarithromycin, sorafenib , methylprednisone, minocycline, dexamethasone, cisplatin, atorvastatin, simvastatin, lovastatin, amiodarone, carbamazepine, carvedilol, chlorpromazine, sisha Bili, dapsone, azithromycin, neomycin, amphotericin B, griseofulvin, celecoxib, raloxifene, flurbiprofen, indomethacin, ibuprofen, tamoxifen Fen, diclofenac, naproxen, piroxicam, ralivide, efavirenz, nelfinavir, atazanavir, ritonavir, sirolimus, spironolactone, tacrolimus , helinolol, terfenadine, estradiol, vitamin A, vitamin D, vitamin E, vitamin K, cyclosporine or insulin One or a mixture of several.
The method for preparing ultra-high drug-loaded nanoparticles according to the sequence precipitation complex coacervation method according to claim 1, wherein the first outer shell material is chitosan, and the second outer shell material is alginic acid. Sodium, hyaluronic acid, heparin, poly-2-acrylamide-2-methylpropane, kondagogu gum, pectin, xanthan gum, poly-D-glutamic acid, dextran sulfate, carrageenan, carboxymethyl cellulose Or sodium carboxymethyl cellulose;

Alternatively, the first outer shell material is polyethyleneimine and its derivatives, spermine and its derivatives, polylysine and its derivatives, polyurethane and its derivatives, cationic cyclodextrin and its derivatives. Or a cationic phospholipid, or a cationic dextran and a derivative thereof, wherein the second outer shell material is deoxyribonucleic acid or ribonucleic acid;

Alternatively, the first outer shell material is polystyrene sulfonate, and the second outer shell material is polydiallyl dimethyl;

Or the first outer shell material is carboxymethyl cellulose, and the second outer shell material is N-methylated polyvinyl pyridine;

Alternatively, the first outer casing material is polyvinyl alcohol sulfuric acid, and the second outer casing material is polydiallyldimethylammonium chloride;

Alternatively, the first outer casing material is Eudragit E, and the second outer casing material is carrageenan;

Or the first outer shell material is polymethacrylic acid, and the second outer shell material is poly N-ethyl-4-vinylpyridine bromide;

Alternatively, the first outer shell material is albumin, and the second outer shell material is dextran sulfate, sodium carboxymethyl cellulose, alginate, pectate, pectin, cationic polyglutamine. Acid or dextran;

Alternatively, the first outer shell material is β-lactoglobulin, and the second outer shell material is sodium carboxymethyl cellulose, sulfated cellulose, carrageenan, guar gum, glutamic acid Sugar, gum arabic, dextran sulfate or propylene glycol alginate.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 1, wherein the solvent I is water, or a buffer solution, or an aqueous solution containing an organic solvent; II is an organic solvent; the solvent III is water or a buffer.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 6, wherein the buffer solution comprises, but not limited to, a hydrochloride buffer solution, a borate buffer solution, and a nitrate salt. Buffer, sulfate buffer, phosphate buffer, citrate buffer, carbonate buffer, acetate buffer, barbiturate buffer, Tris (tris) buffer , 2-(N-morpholine) ethanesulfonic acid buffer, hydroxyethylpiperazine ethanesulfuric acid buffer, ammonium chloride buffer, ethylenediamine buffer or triethylamine buffer.
The method for preparing ultra-high drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 6, wherein the organic solvent comprises, but not limited to, methanol, ethanol, ethylene glycol, diethylene glycol, and different Propanol, 1-propanol, 1,2-propanediol, 1,3-propanediol, butanol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 1,5 - pentanediol, 2-butoxyethanol, glycerol, methyldiethanolamine, diethanolamine, acetone, acetonitrile, diethylenetriamine, dimethoxyethane, ethylamine, dimethyl sulfoxide, dimethyl A mixture of any one or more of formamide, tetrahydrofuran, acetaldehyde, pyridine, triethylene glycol, ethyl acetate, dimethyl carbonate, dichloromethane, cyclohexane, n-octanol or chloroform.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 6, wherein the organic solvent concentration in the aqueous solution containing the organic solvent is in the range of 10 to 40% (v/ v).
The method according to claim 1, wherein the first reactor has a residence time of 0.01 s to 500 s.
The method according to claim 1, wherein the Reynolds number of the entire reaction system is from 1 to 2,000.
A method for preparing ultrahigh drug-loaded nanoparticles by a sequence precipitation complexation coacervation method, characterized in that firstly, the first reactant and the second reactant are mixed in the first reactor, so that the active drug component is in the first reaction The drug nanoparticle core is rapidly precipitated in the device; the reaction product of the first reactor rapidly flows into the second reactor, is mixed with the third reactant, and the shell material is complexed and agglomerated to deposit on the surface of the drug nanoparticle core. Forming ultra-high drug-loaded nanoparticles having a core-shell structure;

among them,

The first reactant is a solution formed by both the first outer shell material and the solvent IV, and the solvent IV is a poor solvent of the active pharmaceutical ingredient and is a benign solvent of the first outer shell material;

The second reactant is a solution formed by both the active pharmaceutical ingredient and the solvent V;

The third reactant is a solution formed by the second outer shell material and the solvent VI, wherein the solvent VI is a poor solvent of the active pharmaceutical component and is a benign solvent of the first outer shell material and the second outer shell material;

The solvents IV, V and VI are mutually soluble;

The first outer shell material and the second outer shell material can undergo a complex condensation reaction;

In the first reactor, the incoming flow rate of the first reactant is greater than the incoming flow rate of the second reactant.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 12, characterized in that the ultrahigh drug-loaded nanoparticles comprise a nano-core containing an active pharmaceutical ingredient and control drug release The outer shell; wherein the mass of the active pharmaceutical ingredient accounts for 30-85% of the mass of the whole nanoparticle; and the ultrahigh drug-loaded nanoparticle has a particle diameter of 50-2000 nm.
The method according to claim 12, wherein the active pharmaceutical ingredient is a poorly water-soluble drug.
The method according to claim 14, wherein the poorly water-soluble drug comprises, but is not limited to, paclitaxel, docetaxel, doxorubicin , vincristine, camptothecin, hydroxycamptothecin, etoposide, curcumin, retinoic acid, fluorouracil, methotrexate, teniposide, daunorubicin, aclarithromycin, sorafenib , methylprednisone, minocycline, dexamethasone, cisplatin, atorvastatin, simvastatin, lovastatin, amiodarone, carbamazepine, carvedilol, chlorpromazine, sisha Bili, dapsone, azithromycin, neomycin, amphotericin B, griseofulvin, celecoxib, raloxifene, flurbiprofen, guanidine Mexin, ibuprofen, tamoxifen, diclofenac, naproxen, piroxicam, ralivide, efavirenz, nelfinavir, atazanavir, ritonavir, sirolimus Any one or more of spironolactone, tacrolimus, lincolol, terfenadine, estradiol, vitamin A, vitamin D, vitamin E, vitamin K, cyclosporine or insulin mixture.
The method for preparing ultra-high drug-loaded nanoparticles according to the sequence precipitation complexation coagulation method according to claim 12, wherein the first outer shell material is chitosan, and the second outer shell material is alginic acid. Sodium, hyaluronic acid, heparin, poly-2-acrylamide-2-methylpropane, kondagogu gum, pectin, xanthan gum, poly-D-glutamic acid, dextran sulfate, carrageenan, carboxymethyl cellulose Or sodium carboxymethyl cellulose;

Alternatively, the first outer shell material is polyethyleneimine and its derivatives, spermine and its derivatives, polylysine and its derivatives, polyurethane and its derivatives, cationic cyclodextrin and its derivatives. Or a cationic phospholipid, or a cationic dextran and a derivative thereof, wherein the second outer shell material is deoxyribonucleic acid or ribonucleic acid;

Alternatively, the first outer shell material is polystyrene sulfonate, and the second outer shell material is polydiallyl dimethyl;

Or the first outer shell material is carboxymethyl cellulose, and the second outer shell material is N-methylated polyvinyl pyridine;

Alternatively, the first outer casing material is polyvinyl alcohol sulfuric acid, and the second outer casing material is polydiallyldimethylammonium chloride;

Alternatively, the first outer casing material is Eudragit E, and the second outer casing material is carrageenan;

Or the first outer shell material is polymethacrylic acid, and the second outer shell material is poly N-ethyl-4-vinylpyridine bromide;

Alternatively, the first outer shell material is albumin, and the second outer shell material is dextran sulfate, sodium carboxymethyl cellulose, alginate, pectate, pectin, cationic polyglutamine. Acid or dextran;

Alternatively, the first outer shell material is β-lactoglobulin, and the second outer shell material is sodium carboxymethyl cellulose, sulfated cellulose, carrageenan, guar gum, glutamic acid Sugar, gum arabic, dextran sulfate or propylene glycol alginate.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 12, wherein the solvent IV is water, or a buffer solution, or an aqueous solution containing an organic solvent; V is an organic solvent; the solvent VI is water or a buffer.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 17, wherein the buffer solution comprises, but not limited to, a hydrochloride buffer solution, a borate buffer solution, and a nitrate salt. Buffer, sulfate buffer, phosphate buffer, citrate buffer, carbonate buffer, acetate buffer, barbiturate buffer, Tris (tris) buffer , 2-(N-morpholine) ethanesulfonic acid buffer, hydroxyethylpiperazine ethanesulfuric acid buffer, ammonium chloride buffer, ethylenediamine buffer or triethylamine buffer.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 17, wherein the organic solvent comprises, but not limited to, methanol, ethanol, ethylene glycol, diethylene glycol, and different Propanol, 1-propanol, 1,2-propanediol, 1,3-propanediol, butanol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 1,5 - pentanediol, 2-butoxyethanol, glycerol, methyldiethanolamine, diethanolamine, acetone, acetonitrile, diethylenetriamine, dimethoxyethane, ethylamine, dimethyl sulfoxide, dimethyl A mixture of any one or more of formamide, tetrahydrofuran, acetaldehyde, pyridine, triethylene glycol, ethyl acetate, dimethyl carbonate, dichloromethane, cyclohexane, n-octanol or chloroform.
The method for preparing ultrahigh drug-loaded nanoparticles according to the sequence precipitation complexation coacervation method according to claim 17, wherein the organic solvent concentration in the aqueous solution containing the organic solvent is in the range of 10 to 40% (v/ v).
The method according to claim 12, wherein the first reactor has a residence time of 0.01 s to 500 s.
The method for preparing ultrahigh drug-loaded nanoparticles by the sequence precipitation complexation coacervation method according to claim 12, wherein the Reynolds number of the entire reaction system is from 1 to 2,000.