CN114146647A - Continuous flow preparation method of high drug-loading microspheres - Google Patents
Continuous flow preparation method of high drug-loading microspheres Download PDFInfo
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Abstract
The invention belongs to the technical field of pharmaceutical preparations, and discloses a continuous flow preparation method of high drug-loading microspheres. The method comprises the steps of precipitating an active drug solution by using a poor solvent dissolved with microsphere matrix materials to form nanoparticles, directly using the mixture as an oil phase, mixing the oil phase and a water phase to prepare oil-in-water emulsion droplets, and solidifying to obtain the high drug-loading microspheres. The continuous flow preparation directly wraps the nanoparticles in one step, simplifies the preparation process of the microspheres and has higher application value. The prepared high drug-loading microspheres comprise nanoparticles consisting of active drug ingredients and microsphere matrix materials for controlling drug release, and the particle size is 1-2000 μm; the active medicine component accounts for 5-80% of the total microsphere, and the active medicine has an encapsulation rate of 5-100%. The microsphere prepared by the continuous flow method has high encapsulation efficiency and drug-loading rate, has no burst release phenomenon, can effectively improve the treatment efficiency, and reduces toxic and side effects and adverse reactions.
Description
Technical Field
The invention relates to a continuous flow preparation method of high-loading polypeptide and protein microspheres, belonging to the technical field of pharmaceutical preparations.
Background
The polypeptide and protein are one of the important components of organisms, and have the advantages of high activity, high specificity and low toxicity. Since 1982, recombinant human insulin is approved by the U.S. food and drug administration to come into the market, and the development of polypeptide and protein drugs enters the motorway and is favored by researchers at home and abroad. At present, about 80 kinds of polypeptide and protein medicines are used for clinical treatment of diseases globally, more than 150 kinds of polypeptide and protein medicines are in clinical development stage, and more than 600 kinds of polypeptide and protein medicines are in preclinical research. Compared with small molecule drugs, polypeptide and protein drugs occupy a unique position, and the sale amount of the polypeptide and protein drugs in 2019 in the world exceeds 500 hundred million dollars, which accounts for 5 percent of the global drug market.
Compared with small molecular drugs, the polypeptide and protein drugs have complex structures and are sensitive to in vivo protease, physiological temperature and pH value changes, so that the in vivo and in vitro stability is poor, the half-life period is short, the bioavailability is low, and the clinical application of the drugs is limited. The clinically common dosage form of polypeptide and protein medicines is an injection solution or a freeze-dried powder injection, and in order to maintain effective blood concentration, patients need to frequently administer the medicines, so that the compliance is poor. The polypeptide and protein drugs are encapsulated in the microsphere matrix, so that the polypeptide and the protein can be protected from the influence of the external environment, and the stability of the polypeptide and the protein is improved. Meanwhile, the microsphere matrix can control the release rate of the drug and maintain the effective blood concentration for a longer time, thereby reducing the administration frequency and improving the compliance of patients.
The preparation of the polypeptide and protein drugs into injectable microsphere preparations is an ideal administration route. However, the preparation of microsphere preparations still has many places which need to be perfected, and the existing microsphere preparation process generally has the problems of low encapsulation efficiency, limited drug-loading rate of the prepared microspheres, uncontrollable release behavior and the like. The medicine carrying amount is improved, the use of auxiliary materials can be greatly reduced, the production cost is reduced, toxic and side effects are avoided, the curative effect is enhanced, and the compliance of patients is increased. The problem of sudden drug release is often caused by increasing the drug loading rate, so that the blood concentration is increased in a short time, and serious side effects are possibly caused. Therefore, the development of the microsphere with high drug loading capacity and controllable release has very important significance for a microsphere drug delivery system, and the preparation method has important guiding significance for the future development of the field of pharmaceutical preparations.
In addition, the batch preparation method of the traditional microspheres has the defects that the particle size distribution range of the microspheres in the same batch is wide, and the average particle size, the encapsulation efficiency, the drug loading rate and the drug release difference of the microspheres in different batches are large. Continuous flow preparation can not only overcome the difference between batches, but also simplify the production process, shorten the production period, improve the production efficiency and reduce the production cost and risk. The continuous production has incomparable advantages compared with the traditional batch production and represents the development direction of modernization of preparation production. Currently, formulation production is undergoing a revolutionary revolution from batch to continuous production.
Disclosure of Invention
The purpose of the invention is as follows: provides a preparation method of microspheres with high drug loading and controllable drug release.
The technical scheme is as follows: the invention provides a preparation method of high drug-loading microspheres, which comprises the steps of preparing, emulsifying and curing drug nanoparticles by a continuous flow device in one step. The prepared high drug-loading microspheres comprise nanoparticles consisting of active drug ingredients and microsphere matrix materials for controlling drug release; wherein, the mass of the active medicine component accounts for 5 to 80 percent of the mass of the whole microsphere; the encapsulation rate of the active medicine component is 5% -100%; the particle size of the high drug-loaded microsphere is 1-2000 μm.
Further, the continuous flow preparation method of the high drug-loaded microsphere comprises the following steps: mixing a first reactant and a second reactant in a first reactor to obtain a mixed system consisting of active drug nanoparticles and microsphere matrix material solution, wherein the system flows to a second reactor without any treatment and is mixed with a third reactant to prepare an oil-in-water emulsion; and finally solidifying the oil-in-water emulsion.
The first reactant is a solution formed by a solvent I and a microsphere matrix material, and the solvent I is a poor solvent of the active pharmaceutical ingredient and a good solvent of the microsphere matrix material; the second reactant is a solution formed by polypeptide, protein medicine and solvent II; the third reactant is a solution formed by water, an emulsifier and salts;
the solvent I and the solvent II are mutually soluble, and a mixture formed by the first reactant and the second reactant is mutually insoluble with the third reactant.
In the first reactor, the flow rate of the first reactant is larger than that of the second reactant, so as to ensure that the volume ratio of the solvent I to the solvent II is larger than 1:1, so that polypeptide and protein medicines can be completely precipitated to form nanoparticles in the process of mixing the first reactant and the second reactant;
in the second reactor, the third reactant is admitted at a flow rate greater than that of the reaction output of the first reactor to form an oil-in-water emulsion.
Further, the continuous flow preparation method of the high drug-loaded microsphere is characterized in that the solvent I is an organic solvent, and comprises any one or a mixture of several of methanol, ethanol, ethylene glycol, diethylene glycol, isopropanol, 1, 2-propylene glycol, 1, 3-propylene glycol, tert-butyl alcohol, 1, 2-butylene glycol, 1, 3-butylene glycol, 1, 4-butylene glycol, 1, 5-pentanediol, 2-butoxyethanol, methyldiethanolamine, acetone, diethanolamine, acetonitrile, ethylamine, diethylenetriamine, dimethyl sulfoxide, tetrahydrofuran, dimethylformamide, dimethoxyethane, pyridine, acetic acid, acetaldehyde or dioxane.
Further, the continuous flow preparation method of the high drug-loaded microsphere is characterized in that the solvent II is water, an acid-containing aqueous solution, an alkali-containing aqueous solution or an aqueous solution containing an organic solvent.
Further, the continuous flow preparation method of the high drug-loaded microsphere is characterized in that: the acid-containing aqueous solution comprises any one or a mixture of more of aqueous solutions of hydrochloric acid, acetic acid, glacial acetic acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid, oxalic acid, boric acid, perchloric acid, silicic acid, formic acid, propionic acid, malonic acid, succinic acid, oxalic acid, sebacic acid, fumaric acid, bromovaleric acid, trifluoroacetic acid and hydrogen bromide acetic acid; the alkaline aqueous solution comprises any one or a mixture of more of aqueous solutions of sodium hydroxide, potassium hydroxide, ammonia water, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate and ethylenediamine; the organic solvent comprises one or a mixture of more of dimethylformamide and dimethyl sulfoxide.
Further, the continuous flow preparation method of the high drug-loaded microsphere is characterized in that: the emulsifier comprises one or a mixture of more of lauric acid soap, rosin oil soap, oleic acid soap, stearic acid soap, alkyl sulfonate, alkyl naphthyl sulfonate, alkyl sulfate, alkylbenzene sulfonate, lignosulfonate, phosphate ester salt, alkyl ammonium salt, polyvinylpyrrolidone, lecithin, sulfate ester salt, quaternary ammonium salt, fatty glyceride, sucrose fatty acid ester, polyoxyethylene fatty alcohol ether, sorbitan fatty acid, polysorbate, crown ether type surfactant, polyoxyethylene-polyoxypropylene block copolymer, silicon-containing surfactant, fluorocarbon surfactant, biosurfactant, gelatin, apricot gum, Arabic gum, tragacanth gum and yolk or solid particle emulsifier.
Further, the continuous flow preparation method of the high drug-loaded microsphere is characterized in that: the salts comprise sodium chloride, sodium nitrate, sodium sulfate, potassium chloride, potassium nitrate, potassium sulfate, calcium chloride, calcium nitrate, calcium sulfate, ammonium chloride, ammonium nitrate, ammonium sulfate, magnesium chloride, magnesium nitrate, magnesium sulfate, aluminum chloride, aluminum nitrate, aluminum sulfate, zinc sulfate, sodium carbonate, sodium bicarbonate and potassium carbonate; the salt content ranges from 20% to 75% (w/w).
The invention also discloses a continuous flow preparation device of the high drug-loaded microspheres, which is characterized in that: comprises 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 third inlet, a fourth inlet and a second outlet;
the first inlet is used for introducing a first reactant or a second reactant;
the second inlet is used for introducing a second reactant or a first reactant;
the first reactor is used for mixing a first reactant and a second reactant to form active drug nanoparticles;
the first outlet of the first reactor is connected with the third inlet of the second reactor and is used for conveying the mixed product of the first reactant and the second reactant to the second reactor;
the fourth inlet is used for introducing a third reactant;
the second reactor is used for mixing a mixed product of the first reactant and the second reactant with the third reactant to form the oil-in-water emulsion.
The conduit of the second outlet of the second reactor is used for solidification of the oil-in-water emulsion.
Further, the preparation method of the high drug-loaded microsphere is characterized in that: the high drug-loading microspheres comprise nanoparticles consisting of active drug ingredients and microsphere matrix materials for controlling drug release; wherein, the mass of the active medicine component accounts for 5 to 80 percent of the mass of the whole microsphere; the encapsulation rate of the active medicine component is 5% -100%; the particle size of the high drug-loaded microsphere is 1-2000 μm.
Further, the high drug-loading microsphere is characterized in that the active drug ingredients comprise water-soluble polypeptide and protein and a mixture thereof. The microsphere matrix material capable of controlling drug release comprises any one of polymer and lipid material and a mixture thereof.
Further, the high drug-loaded microsphere is characterized in that: the water-soluble polypeptides and proteins include, but are not limited to, insulin, whey protein, bovine serum albumin, lactoglobulin, lactalbumin, histone, protamine, mucin, casein, conalbumin, mucin, ferritin, lactoferrin, N-dimethylcasein, calcitonin, salmon calcitonin, tyrosinapine, Sifuweilin, exenatide, beraprot, liraglutide, Antiargitide, teriparatide, loxapide, dolautide, linatide, nesiritide, eptide, eptifibatide, isomabide, tharpeptide, thaumareuptatide, lixilade, leuprorelin, protarelin, gonadorelin, thymoerelin, buserelin, semarerelin, nafarerelin, lanreotide, octreotide, somatostatin, terlipressin, tosiban, glycopeptides, ginseng, nafarelin, hispids, sarelins, etc, Bone peptide, sarcosin, glutathione, mannatide, enfuvirin, bradykinin, thymopentin, enkephalin, nosiheptide, coleptide, hirudin, glatiramer acetate, bee venom, frog venom, aprotinin, alanrelin, pancreatin peptidase, polymyxin, triptorelin, temorelin, serrapeptase, glucagon, vasopressin, histamine, adenohypophysin, thyroid hormone releasing hormone, parathyroid hormone, papain, trypsin, rennin, chymosin, anaplase, fibrinolysin, pancreatic deoxyribonuclease, thrombin, penicillinase, bacitracin, glatiramer, gratin, vancomycin, norvancomycin, teicoplanin, amylase, cellulase, pepsin, pancreatin.
Further, the high drug-loaded microsphere is characterized in that: the polymer comprises any one or a mixture of more of hydrophobic chitosan and derivatives thereof, acetalized dextran and derivatives thereof, polylactic acid and derivatives thereof, polyorthoester and derivatives thereof, polylactic acid-glycolic acid copolymer and derivatives thereof, polyethylene glycol-polylactic acid-glycolic acid block copolymer and derivatives thereof, polyethylene glycol-polyorthoester and derivatives thereof, polycaprolactone and derivatives thereof, hydroxypropyl methylcellulose phthalate and derivatives thereof, hydroxypropyl methylcellulose acetate succinate and derivatives thereof, poly (N-isopropyl acrylamide) and analogues and derivatives thereof; the lipid material comprises one or more of glyceride and its derivatives, fatty acid and its derivatives, steroid materials and its derivatives, waxy materials and its derivatives, and phospholipid materials and its derivatives.
Advantageous effects
The invention avoids the leakage of polypeptide and protein medicines in the preparation process, realizes the high-efficiency encapsulation of the polypeptide and protein medicines, greatly improves the drug-loading rate of the polypeptide and protein medicines in the microspheres, reduces the dosage of carrier materials, can effectively reduce the production cost, avoids toxic and side effects and enhances the treatment effect. Meanwhile, the phenomenon of burst release of the high drug-loaded microspheres is effectively avoided, the controlled release of the drug is better realized, and the adverse reaction caused by overhigh blood drug concentration can be effectively avoided. The preparation process of the invention only relates to one organic solvent, which can reduce the potential toxicity risk caused by using various organic solvents, and meanwhile, the continuous flow preparation can avoid the purification and concentration of polypeptide and protein drug nanoparticles, directly realize the high-efficiency packaging of the polypeptide and protein nanoparticles, improve the preparation efficiency, compress the production period, reduce the production cost and risk and realize large-batch industrial production. In addition, the continuous flow preparation method can also avoid property difference between microsphere batches so as to ensure the controllability of microsphere quality.
Drawings
Fig. 1, schematic diagram of continuous flow preparation device of ultra-high drug-loaded microspheres of example 1.
Fig. 2, schematic diagram of continuous flow preparation device of ultra-high drug-loaded microspheres of example 2.
Fig. 3, schematic diagram of continuous flow preparation device of ultra-high drug-loaded microspheres of example 3.
Fig. 4, schematic diagram of continuous flow preparation device of ultra-high drug-loaded microspheres of example 4.
Fig. 5, schematic diagram of continuous flow preparation device of ultra-high drug-loaded microspheres of example 5.
FIG. 6 is an optical microscope photograph of ADS-encapsulated Insulin microspheres (Insulin @ ADS) prepared under the condition that the mass ratio of Insulin (Insulin) to spermine-modified Acetalized Dextran (ADS) is 1: 1;
FIG. 7 is an optical microscope photograph of BSA (BSA @ ADS) microspheres coated with spermine modified Acetalized Dextran (ADS) prepared under the condition that the mass ratio of Bovine Serum Albumin (BSA) to spermine modified Acetalized Dextran (ADS) is 1: 1;
FIG. 8 is an optical microscope photograph of β -lactoglobulin (β -LG) and spermine-modified Acetalized Dextran (ADS) microspheres coated with the ADS (β -LG @ ADS) prepared under the condition that the mass ratio of β -lactoglobulin (β -LG) to spermine is 1: 1;
FIG. 9 is a scanning electron micrograph of spermine-modified acetalized dextran-coated insulin microspheres (insulin @ ADS), spermine-modified acetalized dextran-coated bovine serum albumin microspheres (BSA @ ADS), and spermine-modified acetalized dextran-coated beta-lactoglobulin microspheres (beta-LG @ ADS);
FIG. 10 shows encapsulation efficiencies (n 10) of spermine-modified acetalized dextran-coated insulin microspheres (insulin @ ADS), spermine-modified acetalized dextran-coated bovine serum albumin microspheres (BSA @ ADS), and spermine-modified acetalized dextran-coated beta-lactoglobulin microspheres (beta-LG @ ADS).
Fig. 11 shows drug loading (n ═ 10) of spermine-modified acetalized dextran-coated insulin microspheres (insulin @ ADS), spermine-modified acetalized dextran-coated bovine serum albumin microspheres (BSA @ ADS), and spermine-modified acetalized dextran-coated beta-lactoglobulin microspheres (beta-LG @ ADS).
FIG. 12 shows the particle size (n ═ 10) of spermine-modified acetalized dextran-coated insulin microspheres (insulin @ ADS), spermine-modified acetalized dextran-coated bovine serum albumin microspheres (BSA @ ADS), and spermine-modified acetalized dextran-coated beta-lactoglobulin microspheres (beta-LG @ ADS).
FIG. 13 shows the particle size polydispersity (n ═ 10) of spermine-modified acetalized dextran-coated insulin microspheres (insulin @ ADS), spermine-modified acetalized dextran-coated bovine serum albumin microspheres (BSA @ ADS), and spermine-modified acetalized dextran-coated beta-lactoglobulin microspheres (beta-LG @ ADS).
FIG. 14, in vitro release profile of spermine modified acetalized dextran coated insulin microspheres (Insulin @ ADS).
Detailed Description
The invention will be better understood from the following examples. The description of the embodiments is intended to be illustrative, and not to limit the invention described in detail in the claims.
Example 1
As shown in fig. 1, this embodiment discloses a continuous flow preparation device of high drug-loaded microspheres, which includes a first reactor and a second reactor, wherein the first reactor includes a first inlet, a second inlet and a first outlet, and the second reactor includes a third inlet and a fourth inlet.
Example 2
As shown in fig. 2, this example discloses a continuous flow preparation device of high drug-loaded microspheres. The difference between the present embodiment and embodiment 1 is that the second inlets are two, and are respectively perpendicular to the straight line formed by the first reactor and the first inlet; and two fourth inlets are arranged and are respectively vertical to a straight line formed by the second reactor and the third inlet.
The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet respectively, the obtained mixture of the first reactant and the second reactant enters the second reactor through the third inlet under the action of the pump, meanwhile, the third reactant enters the second reactor through the fourth inlet, and the obtained oil-in-water emulsion is solidified to obtain the high drug-loaded microspheres. The exchange of the first reactant and the second reactant into the inlet of the first reactor does not affect the preparation of the high drug-loaded microspheres. The middle dashed part indicates that the length of the reactor can be adjusted.
Example 3
As shown in fig. 3, this example discloses a continuous flow preparation device of high drug-loaded microspheres. The difference between this embodiment and embodiment 2 is that there is one second inlet, and the first inlet and the second inlet both form an included angle (0-180 degrees) with the first reactor, 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, the obtained mixture of the first reactant and the second reactant enters the second reactor through the third inlet under the action of the pump, meanwhile, the third reactant enters the second reactor through the fourth inlet, and the obtained oil-in-water emulsion is solidified to obtain the high drug-loaded microspheres. The exchange of the first reactant and the second reactant into the inlet of the first reactor does not affect the preparation of the high drug-loaded microspheres. The middle dashed part indicates that the length of the reactor can be adjusted.
Example 4
As shown in fig. 4, this example discloses a continuous flow preparation device of high drug-loaded microspheres. The present embodiment is different from embodiment 3 in that there is one fourth inlet, the third inlet and the fourth inlet both form an included angle (0-180 degrees) with the second reactor, 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, the obtained mixture of the first reactant and the second reactant enters the second reactor through the third inlet under the action of the pump, meanwhile, the third reactant enters the second reactor through the fourth inlet, and the obtained oil-in-water emulsion is solidified to obtain the high drug-loaded microspheres. The exchange of the first reactant and the second reactant into the inlet of the first reactor does not affect the preparation of the high drug-loaded microspheres. The middle dashed part indicates that the length of the reactor can be adjusted.
Example 5
As shown in fig. 5, this example discloses a continuous flow preparation device of high drug-loaded microspheres. This example differs from examples 1-4 in that the first reactant and the second reactant are mixed by agitation in the first reactor to form a mixture, which is then mixed with the third reactant by agitation after the mixture enters the second reactor.
The first reactant and the second reactant enter the first reactor through the first inlet and the second inlet respectively, the obtained mixture of the first reactant and the second reactant enters the second reactor through the third inlet under the action of the pump, meanwhile, the third reactant enters the second reactor through the fourth inlet, and the obtained oil-in-water emulsion is solidified to obtain the high drug-loaded microspheres. The exchange of the first reactant and the second reactant into the inlet of the first reactor does not affect the preparation of the high drug-loaded microspheres. The middle dashed part indicates that the length of the reactor can be adjusted.
Example 6
The embodiment discloses preparation of a high-polypeptide and protein drug-loaded microsphere and an Insulin (Insulin) microsphere wrapped by spermine modified Acetalized Dextran (ADS).
Acetone solution of ADS as a first reactant, hydrochloric acid solution of Insulin as a second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium chloride hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres. The encapsulation efficiency of the resulting microspheres was 91.1% (fig. 10), the drug loading was 45.6% (fig. 11), the average particle size was 24.5 μm (fig. 12), and the polydispersity was 4.9% (fig. 13).
Example 7
The embodiment discloses preparation of high-polypeptide and protein drug-loaded microspheres and spermine-modified Acetalized Dextran (ADS) -coated Bovine Serum Albumin (BSA) microspheres.
An acetonitrile solution of ADS as a first reactant, an aqueous solution of BSA as a second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium chloride hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres. The encapsulation efficiency of the resulting microspheres was 97.8% (fig. 10), the drug loading was 48.9% (fig. 11), the average particle size was 34.0 μm (fig. 12), and the polydispersity was 4.2% (fig. 13).
Example 8
The embodiment discloses a preparation method of beta-lactoglobulin (beta-LG) microspheres wrapped by high-polypeptide and protein drug-loaded microspheres and spermine-modified Acetalized Dextran (ADS).
A tetrahydrofuran solution of ADS as a first reactant, an aqueous solution of β -LG as a second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium chloride hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres. The encapsulation efficiency of the resulting microspheres was 96.0% (fig. 10), the drug loading was 48.0% (fig. 11), the average particle size was 28.8 μm (fig. 12), and the polydispersity was 3.9% (fig. 13).
Example 9
The embodiment discloses preparation of a high-polypeptide and protein drug-loaded microsphere, namely an Exenatide (EXT) microsphere wrapped by spermine modified Acetalized Dextran (ADS).
An acetonitrile solution of ADS as a first reactant, an aqueous solution of EXT as a second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium chloride hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres.
Example 10
This example discloses a preparation method of high drug-loading microsphere, polylactic acid-glycolic acid copolymer (PLGA) coated Calcitonin (CT) microsphere.
An acetone solution of PLGA was used as the first reactant, an aqueous solution of calcitonin was used as the second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium chloride hexahydrate (w/w) was used as the third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres.
Example 11
This example discloses the preparation of a high drug loading microsphere, a parathyroid hormone (PTH) microsphere coated with poly (lactic-co-glycolic acid) (PLGA).
An ethanol solution of PLGA as a first reactant, an aqueous solution of PTH as a second reactant, and a mixed solution of 1% pluronic F127 and 60% magnesium chloride hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres.
Example 12
The embodiment discloses a preparation method of a high drug-loading microsphere, namely a Vancomycin (VCM) microsphere coated by polylactic-co-glycolic acid (PLGA).
A methanol solution of PLGA was used as a first reactant, an aqueous solution of VCM was used as a second reactant, and a mixed solution of 2% pluronic F127 and 60% magnesium sulfate hexahydrate (w/w) was used as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres.
Example 13
This example discloses the preparation of a high drug-loaded microsphere, hydroxypropyl methylcellulose acetate succinate (HPMCAS) -coated Buserelin (BA) microspheres.
An acetonitrile solution of HPMCAS as a first reactant, an aqueous solution of BA as a second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium sulfate hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres.
Example 14
This example discloses the preparation of a high drug-loaded microsphere, hydroxypropyl methylcellulose acetate succinate (HPMCAS) -encapsulated Thymopentin (TPP) microspheres.
A tetrahydrofuran solution of HPMCAS as a first reactant, an aqueous solution of TPP as a second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium chloride hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres.
Example 15
This example discloses the preparation of high drug loaded microspheres polylactic acid (PLA) -coated Nafarelin (NAF) microspheres.
An acetonitrile solution of PLA as a first reactant, an aqueous solution of NAF as a second reactant, and a mixed solution of 2% polyvinyl alcohol (PVA) and 60% magnesium chloride hexahydrate (w/w) as a third reactant. In a micro-fluidic device, a first reactant and a second reactant are mixed, a third reactant is added and mixed to obtain an emulsion-in-water emulsion, and finally emulsion droplets are solidified through solvent diffusion to obtain the microspheres.
Examples 16 to 35
Other experimental methods and parameters were the same as in example 1, and the specific raw material selection is shown in table 1.
TABLE 1
Claims (10)
1. A continuous flow preparation device of high medicine carrying microsphere is characterized in that: comprises 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 third inlet, a fourth inlet and a second outlet;
the first inlet is used for introducing a first reactant or a second reactant;
the second inlet is used for introducing a second reactant or a first reactant;
the first reactor is used for mixing a first reactant and a second reactant to form active drug nanoparticles;
the first outlet of the first reactor is connected with the third inlet of the second reactor and is used for conveying the mixed product of the first reactant and the second reactant to the second reactor;
the fourth inlet is used for introducing a third reactant;
the second reactor is used for mixing a mixed product of the first reactant and the second reactant with the third reactant to form an oil-in-water emulsion;
the conduit of the second outlet of the second reactor is used for solidification of the oil-in-water emulsion.
2. A continuous flow preparation method of high drug-loaded microspheres comprises the following steps: the device of claim 1, wherein the first reactant and the second reactant are mixed in the first reactor to obtain a mixed system consisting of active drug nanoparticles and microsphere matrix material solution, and the mixed system flows to the second reactor without any treatment and is mixed with the third reactant to prepare an oil-in-water emulsion; finally, solidifying the oil-in-water emulsion;
the first reactant is a solution formed by a solvent I and a microsphere matrix material, and the solvent I is a poor solvent of the active pharmaceutical ingredient and a good solvent of the microsphere matrix material; the second reactant is a solution formed by polypeptide, protein medicine and solvent II; the third reactant is a solution formed by water, an emulsifier and salts;
the solvent I and the solvent II are mutually soluble, and a mixture formed by the first reactant and the second reactant is mutually insoluble with the third reactant;
in the first reactor, the flow rate of the first reactant is larger than that of the second reactant, so as to ensure that the volume ratio of the solvent I to the solvent II is larger than 1:1, so that polypeptide and protein medicines can be completely precipitated to form nanoparticles in the process of mixing the first reactant and the second reactant;
in the second reactor, the third reactant is admitted at a flow rate greater than that of the reaction output of the first reactor to form an oil-in-water emulsion.
3. The method for preparing microspheres loaded with drugs according to claim 2, wherein the solvent I is an organic solvent, and comprises one or more of methanol, ethanol, ethylene glycol, diethylene glycol, isopropanol, 1, 2-propanediol, 1, 3-propanediol, tert-butanol, 1, 2-butanediol, 1, 3-butanediol, 1, 4-butanediol, 1, 5-pentanediol, 2-butoxyethanol, methyldiethanolamine, acetone, diethanolamine, acetonitrile, ethylamine, diethylenetriamine, dimethyl sulfoxide, tetrahydrofuran, dimethylformamide, dimethoxyethane, pyridine, acetic acid, acetaldehyde or dioxane.
4. The preparation method of the high drug-loaded microsphere according to claim 2, wherein the solvent II is water, an acid-containing aqueous solution, an alkali-containing aqueous solution or an organic solvent-containing aqueous solution.
5. The preparation method of the high drug-loaded microsphere according to claim 4, wherein the preparation method comprises the following steps: the acid-containing aqueous solution comprises any one or a mixture of more of aqueous solutions of hydrochloric acid, acetic acid, glacial acetic acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid, oxalic acid, boric acid, perchloric acid, silicic acid, formic acid, propionic acid, malonic acid, succinic acid, oxalic acid, sebacic acid, fumaric acid, bromovaleric acid, trifluoroacetic acid and hydrogen bromide acetic acid; the alkaline aqueous solution comprises any one or a mixture of more of aqueous solutions of sodium hydroxide, potassium hydroxide, ammonia water, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate and ethylenediamine; the organic solvent comprises one or a mixture of more of dimethylformamide and dimethyl sulfoxide.
6. The preparation method of the high drug-loaded microsphere according to claim 2, wherein the preparation method comprises the following steps: the emulsifier comprises one or a mixture of more of lauric acid soap, rosin oil soap, oleic acid soap, stearic acid soap, alkyl sulfonate, alkyl naphthyl sulfonate, alkyl sulfate, alkylbenzene sulfonate, lignosulfonate, phosphate ester salt, alkyl ammonium salt, polyvinylpyrrolidone, lecithin, sulfate ester salt, quaternary ammonium salt, fatty glyceride, sucrose fatty acid ester, polyoxyethylene fatty alcohol ether, sorbitan fatty acid, polysorbate, crown ether type surfactant, polyoxyethylene-polyoxypropylene block copolymer, silicon-containing surfactant, fluorocarbon surfactant, biosurfactant, gelatin, apricot gum, Arabic gum, tragacanth gum and yolk or solid particle emulsifier.
7. The preparation method of the high drug-loaded microsphere according to claim 2, wherein the preparation method comprises the following steps: the salts comprise sodium chloride, sodium nitrate, sodium sulfate, potassium chloride, potassium nitrate, potassium sulfate, calcium chloride, calcium nitrate, calcium sulfate, ammonium chloride, ammonium nitrate, ammonium sulfate, magnesium chloride, magnesium nitrate, magnesium sulfate, aluminum chloride, aluminum nitrate, aluminum sulfate, zinc sulfate, sodium carbonate, sodium bicarbonate and potassium carbonate; the salt content ranges from 20% to 75% w/w.
8. The preparation method of the high drug-loaded microsphere according to claim 2, wherein the preparation method comprises the following steps: the high drug-loading microspheres comprise nanoparticles consisting of active drug ingredients and microsphere matrix materials for controlling drug release; wherein, the mass of the active medicine component accounts for 5 to 80 percent of the mass of the whole microsphere; the encapsulation rate of the active medicine component is 5% -100%; the particle size of the high drug-loaded microsphere is 1-2000 μm.
9. The preparation method of the high drug-loaded microsphere according to claim 8, wherein the active pharmaceutical ingredient comprises water-soluble polypeptide and protein and a mixture thereof; the microsphere matrix material capable of controlling drug release comprises any one of polymer and lipid material and a mixture thereof.
10. The high drug-loaded microsphere of claim 8, wherein: the water-soluble polypeptides and proteins include, but are not limited to, insulin, whey protein, bovine serum albumin, lactoglobulin, lactalbumin, histone, protamine, mucin, casein, conalbumin, mucin, ferritin, lactoferrin, N-dimethylcasein, calcitonin, salmon calcitonin, tyrosinapine, Sifuweilin, exenatide, beraprot, liraglutide, Antiargitide, teriparatide, loxapide, dolautide, linatide, nesiritide, eptide, eptifibatide, isomabide, tharpeptide, thaumareuptatide, lixilade, leuprorelin, protarelin, gonadorelin, thymoerelin, buserelin, semarerelin, nafarerelin, lanreotide, octreotide, somatostatin, terlipressin, tosiban, glycopeptides, ginseng, nafarelin, hispids, sarelins, etc, Bone peptide, sarcosin, glutathione, mannatide, enfuvirin, bradykinin, thymopentin, enkephalin, nosiheptide, colestipol, hirudin, glatiramer acetate, bee venom, frog venom, aprotinin, alanrelin, pancreatin peptidase, polymyxin, triptorelin, temorelin, serrapeptase, glucagon, vasopressin, histamine, adenohypophyseal hormone, thyroid hormone releasing hormone, parathyroid hormone, papain, trypsin, rennin, chymosin, anaplase, fibrinolysin, pancreatic deoxyribonuclease, thrombin, penicillinase, bacitracin, glatiramer, gratin, vancomycin, norvancomycin, teicoplanin, amylase, cellulase, pepsin, pancreatin;
the polymer comprises any one or a mixture of more of hydrophobic chitosan and derivatives thereof, acetalized dextran and derivatives thereof, polylactic acid, polyorthoester, polylactic acid-glycolic acid copolymer, polyethylene glycol-polylactic acid-glycolic acid block copolymer, polyethylene glycol-polyorthoester, polycaprolactone, hypromellose phthalate, hypromellose acetate succinate, poly (N-isopropylacrylamide) and derivatives; the lipid material comprises one or more of glyceride and its derivatives, fatty acid, steroid material, waxy material and phospholipid material.
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