CN113171356B - Preparation method of sitagliptin drug nano-preparation with stable release performance - Google Patents
Preparation method of sitagliptin drug nano-preparation with stable release performance Download PDFInfo
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Abstract
The invention discloses a preparation method of a sitagliptin drug nano-preparation with stable release performance, which uses dopamine hydrochloride as a packaging material and pi bond accumulation for self-assembly to construct nano-preparation particles, wherein ellagic acid is innovatively introduced as a stabilizer in the preparation process of the preparation particles. Firstly, adding a mixed solution of the pluronic and the sitagliptin into a Tris-HCl solution which is stirred at a high speed, and carrying out ultrasonic treatment; and respectively adding the dopamine hydrochloride solution and the ellagic acid solution, carrying out ultrasonic treatment, carrying out light-proof rotary reaction on the solution at room temperature for 48-72 hours, removing impurities through centrifugation and dialysis, and finally, fixing the volume by using normal saline to obtain the sitagliptin nano preparation solution with the required concentration. The nano preparation prepared by the invention has uniform and stable particles, has very strong in vivo long circulation and slow release effects, has more controllable preparation process due to the introduction of ellagic acid, has stable and uniform product quality, and can fully assist the clinical application of the sitagliptin medicines.
Description
Technical Field
The invention belongs to the technical field of nano preparation, and particularly relates to a preparation method of a sitagliptin drug nano preparation with stable release performance.
Background
Various types of sitagliptin drugs play an important role in the clinical treatment of antidepressants at present, such as fluoxetine (fluxetine), paroxetine (paroxetine), duloxetine (duloxetine), vortioxetine (vortioxetine), reboxetine (reboxetine) and sonchixetine (nisoxepine); atomoxetine (atomoxetine) plays an effective therapeutic role in addition to depression in adolescents or in the accepted attention deficit hyperactivity disorder. The above mentioned related to the single isomers of the sitagliptin molecules all having chiral carbon atoms. Most of the different corresponding isomers of the sitagliptin medicines containing chiral molecules have different pharmacological activities, metabolic processes and toxicological effects after entering human bodies. Compared with other medicines, the sitagliptin medicines have smaller toxic and side effects, but still exist. Common side effects during antidepressant treatment include constipation, dry mouth, nausea, emesis, dizziness, fatigue, stomach discomfort, appetite decrease, mood swings, sexual side effects, difficulty in urination, etc. Especially at the beginning of the treatment and at varying doses, some patients have suicidal consciousness, liver injury and serious cardiovascular risks. Patients who use the sitagliptin medicines generally need longer treatment courses, and if the patients receive less times of administration and maintain more stable blood concentration for a long time, the patients have better effect and compliance on the use of the medicines. Therefore, the long-acting slow release technology has important significance for the application of the sitagliptin medicines.
The nano-drug carrier technology which is developed at high speed in recent years brings new development opportunities for the improvement of clinical medication. By loading the nano preparation, the stability, biocompatibility, metabolizing property and targeting property of the drug molecules in vivo are greatly improved, so that the bioavailability of the drug molecules is improved, and the toxic and side effects are greatly reduced. Through decades of development, the technology of nano-drug carrier has been successful, so tens of nano-drug preparations are currently in clinical application worldwide, and a large number of nano-drugs and carrier systems are in preclinical and clinical trial stages.
Dopamine (dopamine) can be accumulated through pi bonds and oxidized to generate Polydopamine (PDA) through self-polymerization reaction under alkaline condition, and the PDA also has extremely strong adhesiveness due to the polymerization mode, so that the PDA can be deposited on the surfaces of various organic or inorganic materials, thereby becoming an effective surface bionic coating material, and the application of the PDA is extremely wide, such as patent application Nos. 201410756470.0 and 201410355973.7. The preparation of PDA nano particles with different sizes and specifications can be realized through proper process control, and the PDA nano particles have physicochemical properties consistent with PDA coatings, and the stability, biocompatibility and degradability are reliably ensured. The pi bond stacking effect can also enable PDA to act with molecules with benzene rings, so that co-deposition is realized. This effect allows the PDA to be effectively loaded with a variety of therapeutic molecules, thereby forming drug-loaded nanoparticles, such as 201510777717.1 and 201611154834.3, etc.
However, due to the difference between the dopamine molecules and the drug molecules, the PDA drug-loaded nano-particles with uniform particles, regular morphology and stable properties are often difficult to obtain by direct deposition, and further production and application of the PDA drug-loaded nano-particles are greatly restricted. It is therefore necessary to achieve nanoparticle stability by a specific process. Because the synthesis of PDA drug-loaded nano-particles depends on pi bond accumulation, corresponding stable molecules can be added to realize efficient pi bond accumulation. Through multipartite experiments, the ellagic acid can be used as a stabilizer for synthesizing PDA nano-carriers, is a natural polyphenol substance widely existing in various plant fruits, has good oxidation resistance, anticancer and antibacterial functions, can improve human immunity and reduce occurrence of various diseases, and is widely used in the fields of medicines, foods and the like. The molecular structure of the polyphenol dilactone of ellagic acid can play a very strong stabilizing effect in pi bond accumulation, and can realize stable accumulation of PDA and drug molecules. At present, the invention report of using ellagic acid as a stabilizer for synthesizing PDA drug-carrying nano particles is not seen.
Disclosure of Invention
The invention aims to provide a preparation method of a sitagliptin drug nano-preparation with stable release performance, and the prepared nano-preparation has uniform and stable particles, has very strong in vivo long circulation and slow release effects, and can fully assist clinical application of the sitagliptin drug.
The technical scheme adopted by the invention is that the preparation method of the sitagliptin drug nano-preparation with stable release performance is implemented according to the following steps:
step 5, taking out the mixed solution obtained in the step 4, performing ultrasonic treatment, and finally performing light-shielding rotary reaction on the solution at room temperature for 48-72 hours;
The present invention is also characterized in that,
the step 1 is specifically as follows:
Tris-HCl mother liquor: the components are Tris-HCl:3% w/v of pure water as solvent;
the mother liquor of the sitagliptin medicine: the components are sitagliptin medicines, 2% w/v, and the solvent is dimethyl sulfoxide;
pluronic mother liquor: the component is pluronic, 2% w/v, and the solvent is dimethyl sulfoxide;
dopamine hydrochloride mother liquor: the components are dopamine hydrochloride, 2% w/v, and the solvent is pure water;
ellagic acid mother liquor: the component is ellagic acid, 1% w/v, and the solvent is dimethyl sulfoxide;
all mother solutions are treated by ultrasonic treatment to fully dissolve the solvent, and then filtered to remove impurities.
The pH of the mother liquor with the components of Tris-HCl in the step 1 is 8.5-8.8.
In the step 1, when the component is a sitagliptin drug, the methyl sulfoxide in the mother liquor is any one of fluoxetine, tomoxetine, duloxetine, vortioxetine, reboxetine, paroxetine and sony sitagliptin;
pronike is any one of F68, F108 and F127.
In the step 2, the stirring speed of the Tris-HCl mother solution is 1000-1800 revolutions per minute, the dripping speed of the mixed solution of the pluronic solution and the statin drug solution in the Tris-HCl mother solution is 60-120 drops per minute, the stirring speed is reduced to 600-800 revolutions per minute after the dripping is finished, the stirring time is maintained for 10 minutes, the ultrasonic treatment is carried out by using a probe type ultrasonic breaker, the ultrasonic treatment power is 300-600W, the ultrasonic treatment time is 4-6 seconds per cycle, the interval time is 2-3 seconds, and the total cycle number is 20-40 times.
In the step 3, the stirring speed is below 120 r/min, and stirring is maintained for 8-12 hours.
In the step 4, the stirring rotation speed is firstly increased to 400-800 rpm, the dripping speed of the ellagic acid solution is 60-120 drops per minute, the stirring speed is reduced to 120 rpm after dripping, and the stirring time is maintained for 4-8 hours.
And 5, carrying out ultrasonic treatment on the taken mixed solution obtained in the step 4 by using an ultrasonic cleaning machine, wherein the power is 200-250W, the working frequency is 40KHz, the time is 20-30 minutes, and the solution is subjected to rotary reaction at room temperature and in a dark place for 48-72 hours during centrifugal treatment.
In the step 6, the rotational speed of centrifugal impurity removal is 14000 r/min, the centrifugal time is 15 min, the solution is placed in a dialysis bag with the molecular weight cut-off of 10kDa during dialysis, the Tris-HCl solution containing 0.9% NaCl is taken as external solution, and the external solution is dialyzed for 24-48 hours, and the external solution is replaced for 3-5 times during dialysis.
The preparation method of the sitagliptin drug nano-preparation with stable release performance has the beneficial effects that ellagic acid is introduced into the preparation of the PDA drug-carrying nano-preparation as a preparation stabilizer for the first time, so that the problems of uniformity and stability difference of the PDA drug-carrying nano-particles are solved, and the repeatability of quality is ensured. The obtained nano preparation has the particle size smaller than 200 nanometers, has stronger stability, and has no obvious decomposition or agglomeration phenomenon in serum within one month. The entrapment performance is outstanding, no obvious release is seen in one week under normal conditions, and the loaded drug molecules can be continuously and stably released under an acidic environment. The toxicity aspect is also obvious, obvious damage to red blood cells is not shown, and the toxic and side effects of the loaded medicine can be obviously reduced. The core for supporting the series of excellent performances is a PDA drug molecule heterozygous process with ellagic acid as a stabilizer. The technical steps are simple and feasible, and the obtained product can be subjected to accurate quality control and has high stability and releasability.
Drawings
Fig. 1 is a transmission electron micrograph of the fluoxetine nano-formulation prepared in example 1.
Fig. 2 shows the results of three months stability test of the fluoxetine nano-preparation prepared in example 1 in phosphate buffer at 28 ℃.
FIG. 3 shows the results of stability test of the fluoxetine nano-preparation prepared in example 1 in fetal bovine serum at 4deg.C for five weeks.
Fig. 4 shows the results of drug release test of the fluoxetine nano-formulation prepared in example 1 under normal conditions within one week.
Fig. 5 shows the results of drug release test of the fluoxetine nano-formulation prepared in example 1 for 72 hours at different pH conditions.
Fig. 6 shows the results of drug release test of the fluoxetine nano-formulation prepared in example 1 over 72 hours at different temperature conditions.
FIG. 7 hemolysis test result of fluoxetine nanometer preparation prepared in example 1 (pure water is used as positive control, and previous components are respectively used as control)
FIG. 8 shows the results of in vivo acute toxicity tests of fluoxetine nano-formulations prepared in example 1 (note: BALB/c mice were used, weighing 18-22g, each group of male and female halves).
Detailed Description
The invention will be described in detail below with reference to the drawings and the detailed description.
The nanometer preparation of the sitagliptin medicine with stable release performance is prepared by using dopamine hydrochloride as a packing material and an ellagic acid stabilizer. The preparation method comprises the steps of firstly, co-dissolving drug molecules and a small amount of surfactant, namely, pluronic, in dimethyl sulfoxide, dispersing in a weak alkaline Tris-HCl solution, forming drug-loaded nano-micelles in advance, adding dopamine hydrochloride into the solution to enable the dopamine hydrochloride to start to deposit on the surfaces of the micelles, adding ellagic acid after a period of reaction, enabling nano particles to perform stable reaction, and finally removing impurities to obtain the statin drug nano preparation solution with the required concentration. The method is simple and efficient, the prepared nano preparation particles are uniform and stable, have very strong in-vivo long-circulation and slow-release effects, can fully assist clinical application of the sitagliptin medicine, and have very good development prospect.
The invention relates to a preparation method of a sitagliptin drug nano-preparation with stable release performance, which is implemented according to the following steps:
the step 1 is specifically as follows:
Tris-HCl mother liquor: the components are Tris-HCl:3% w/v of pure water as solvent;
the mother liquor of the sitagliptin medicine: the components are sitagliptin medicines, 2% w/v, and the solvent is dimethyl sulfoxide;
pluronic mother liquor: the component is pluronic, 2% w/v, and the solvent is dimethyl sulfoxide;
dopamine hydrochloride mother liquor: the components are dopamine hydrochloride, 2% w/v, and the solvent is pure water;
ellagic acid mother liquor: the component is ellagic acid, 1% w/v, and the solvent is dimethyl sulfoxide;
all mother solutions are treated by ultrasonic treatment to fully dissolve the solvent, and then filtered to remove impurities.
The pH of the mother liquor with the components of Tris-HCl in the step 1 is 8.5-8.8.
In the step 1, when the component is a sitagliptin drug, the methyl sulfoxide in the mother liquor is any one of fluoxetine, tomoxetine, duloxetine, vortioxetine, reboxetine, paroxetine and sony sitagliptin;
pronike is any one of F68, F108 and F127.
in the step 2, the stirring speed of the Tris-HCl mother solution is 1000-1800 revolutions per minute, the dripping speed of the mixed solution of the pluronic solution and the statin drug solution in the Tris-HCl mother solution is 60-120 drops per minute, the stirring speed is reduced to 600-800 revolutions per minute after the dripping is finished, the stirring time is maintained for 10 minutes, the ultrasonic treatment is carried out by using a probe type ultrasonic breaker, the ultrasonic treatment power is 300-600W, the ultrasonic treatment time is 4-6 seconds per cycle, the interval time is 2-3 seconds, and the total cycle number is 20-40 times.
in the step 3, the stirring speed is below 120 r/min, and stirring is maintained for 8-12 hours.
in the step 4, the stirring rotation speed is firstly increased to 400-800 rpm, the dripping speed of the ellagic acid solution is 60-120 drops per minute, the stirring speed is reduced to 120 rpm after dripping, and the stirring time is maintained for 4-8 hours.
Step 5, taking out the mixed solution obtained in the step 4, performing ultrasonic treatment, and finally performing light-shielding rotary reaction on the solution at room temperature for 48-72 hours;
and 5, carrying out ultrasonic treatment on the taken mixed solution obtained in the step 4 by using an ultrasonic cleaning machine, wherein the power is 200-250W, the working frequency is 40KHz, the time is 20-30 minutes, and the solution is subjected to rotary reaction at room temperature and in a dark place for 48-72 hours during centrifugal treatment.
In the step 6, the rotational speed of centrifugal impurity removal is 14000 r/min, the centrifugal time is 15 min, the solution is placed in a dialysis bag with the molecular weight cut-off of 10kDa during dialysis, the Tris-HCl solution containing 0.9% NaCl is taken as external solution, and the external solution is dialyzed for 24-48 hours, and the external solution is replaced for 3-5 times during dialysis.
Example 1
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.5);
fluoxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F127:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 200W and the working frequency of 40KHz for 20 minutes, transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 72 hours;
Example 2
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.8);
tomoxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F68:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 250W and the working frequency of 40KHz for 30 minutes, transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 72 hours;
Example 3
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.6);
duloxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F108:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 220W, the working frequency of 40KHz and the time of 25 minutes, and finally transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 48 hours;
Example 4
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.7);
vortioxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F127:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 240W and the working frequency of 40KHz for 25 minutes, transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 72 hours;
Example 5
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.8);
reboxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F68:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 250W, the working frequency of 40KHz and the time of 0 min, and finally transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 72 hours;
Example 6
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.5);
paroxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F108:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 200W and the working frequency of 40KHz for 20 minutes, transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 48 hours;
Example 7
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.7);
sony sitagliptin: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F127:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 230W and the working frequency of 40KHz for 25 minutes, and finally transferring the solution into a centrifuge tube to perform rotary reaction at room temperature and in a dark place for 72 hours;
Example 8
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.5-8.8);
fluoxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F68:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 200W and the working frequency of 40KHz for 30 minutes, transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 72 hours;
Example 9
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.5-8.8);
reboxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F127:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 220W and the working frequency of 40KHz for 20 minutes, transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 60 hours;
Example 10
The embodiment comprises the following steps:
Tris-HCl:3% w/v, (solvent: pure water, pH adjusted to 8.5-8.8);
paroxetine: 2% w/v, (solvent: dimethyl sulfoxide);
pluronic F108:2% w/v, (solvent: dimethyl sulfoxide);
dopamine hydrochloride: 2% w/v, (solvent: pure water);
ellagic acid: 1% w/v, (solvent: dimethyl sulfoxide).
Fully dissolving all mother liquor by ultrasonic treatment, and filtering to remove impurities by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers according to different solvents;
step 5, taking out the solution from the round bottom flask, performing ultrasonic treatment by using an ultrasonic cleaner with the power of 210W and the working frequency of 40KHz for 25 minutes, transferring the solution into a centrifuge tube, and performing rotary reaction at room temperature and in a dark place for 66 hours;
The morphology, particle size, stability, release and in vivo toxicity of the fluoxetine nanopreparation prepared in example 1 were measured. The obtained results showed that: the nanometer preparation has uniform particles, good monodispersity, strong stability, uniform morphological characteristics, good monodispersity (shown in figure 1), and high stability (shown in figures 2 and 3). The release experiment results show that the nano preparation has good slow release effect, and can ensure the sustained release of the medicine in the systemic circulation (see fig. 4, 5 and 6). The toxicity of the fluoxetine nano-preparation is measured by taking fluoxetine as a control, and the result shows that the hemolysis of the fluoxetine nano-preparation to erythrocytes is very small (see figure 7), and the nano-preparation effectively reduces the dosage form toxicity of the fluoxetine from the aspect of in vivo survival rate change (figure 8).
The most important process core in the invention is that ellagic acid is introduced into the preparation of PDA nano drug-carrying particles as a stabilizer, and the addition of ellagic acid greatly improves the monodispersity and the particle uniformity of nano carriers, and greatly improves the completeness and the repeatability of the preparation process and the accuracy of product quality control. Although a plurality of related PDA drug carrying preparation processes are disclosed, a series of experiments show that the effect of directly carrying the drug by the simple PDA nano particles is not remarkable, and certain drugs need to be directly carried in a targeted manner, or the drugs need to be treated in advance and then the PDA is deposited on the drug to realize the encapsulation. The invention fully overcomes the series of problems after introducing ellagic acid, and obtains a simple, effective and reliable process.
Claims (1)
1. The preparation method of the sitagliptin drug nano-preparation with stable release performance is characterized by comprising the following steps:
step 1, preparing mother liquor of each component, the components are Tris-HCl, sitagliptin, pluronic, dopamine hydrochloride and ellagic acid;
the step 1 specifically comprises the following steps:
Tris-HCl mother liquor: the components are Tris-HCl:3% w/v of pure water as solvent;
the mother liquor of the sitagliptin medicine: the components are sitagliptin medicines, 2% w/v, and the solvent is dimethyl sulfoxide;
pluronic mother liquor: the component is pluronic, 2% w/v, and the solvent is dimethyl sulfoxide;
dopamine hydrochloride mother liquor: the components are dopamine hydrochloride, 2% w/v, and the solvent is pure water;
ellagic acid mother liquor: the component is ellagic acid, 1% w/v, and the solvent is dimethyl sulfoxide;
the mother solution is treated by ultrasonic treatment to fully dissolve the solvent, and then the impurities are removed by filtration;
the pH value of the mother solution with the components of Tris-HCl in the step 1 is 8.5-8.8;
in the step 1, when the component is a sitagliptin drug, the methyl sulfoxide in the mother liquor is any one of fluoxetine, tomoxetine, duloxetine, vortioxetine, reboxetine, paroxetine and sony sitagliptin; the pluronic is any one of F68, F108 and F127;
the step 1 is carried out by using a microporous filter membrane with fat solubility or water solubility of 0.22 micrometers;
step 2, stirring a Tris-HCl mother solution, fully mixing a pluronic solution and a siteritoneum drug solution, dripping the mixed solution of the pluronic solution and the siteritoneum drug solution into the Tris-HCl mother solution, wherein the mass ratio of the pluronic to the siteritoneum drug is 1:2, the volume ratio of the mixed solution of the pluronic to the siteritoneum drug to the Tris-HCl solution is 10:3-50:3, reducing the stirring speed after dripping, and then carrying out ultrasonic treatment on the whole liquid system, wherein an ice bath is kept all the time in the treatment process;
in the step 2, the stirring speed of the Tris-HCl mother solution is 1000-1800 rpm, the dripping speed of the mixed solution of the pluronic solution and the statin solution in the Tris-HCl mother solution is 60-120 drops per minute, the stirring speed is reduced to 600-800 rpm after the dripping is finished, the stirring time is maintained for 10 minutes, the ultrasonic treatment is carried out by using a probe type ultrasonic breaker, the ultrasonic treatment power is 300-600W, the ultrasonic treatment time is 4-6 seconds per cycle, the interval time is 2-3 seconds, and the total cycle number is 20-40 times;
step 3, adding a dopamine hydrochloride solution into the solution obtained in the step 2 according to the proportion of 4:13-4:38, and stirring;
in the step 3, the stirring speed is below 120 r/min, and stirring is maintained for 8-12 hours;
step 4, increasing the stirring rotation speed, dripping the ellagic acid solution into the mixed solution obtained in the step 3 according to the proportion of 1:11-1:25, and reducing the stirring speed after uniform dispersion;
in the step 4, the stirring rotation speed is firstly increased to 400-800 rpm, the dripping speed of the ellagic acid solution is 60-120 drops per minute, the stirring speed is reduced to 120 rpm after dripping, and the stirring time is maintained for 4-8 hours;
step 5, taking out the mixed solution obtained in the step 4, performing ultrasonic treatment, and finally performing light-shielding rotary reaction on the solution at room temperature for 48-72 hours;
performing ultrasonic treatment on the taken mixed solution obtained in the step 4 by using an ultrasonic cleaning machine, wherein the power is 200-250W, the working frequency is 40KHz, the time is 20-30 minutes, and the solution is subjected to centrifugal treatment and is subjected to rotary reaction at room temperature in a dark place for 48-72 hours;
step 6, removing impurities from the solution obtained in the step 5 through centrifugation, and then dialyzing the solution to remove dimethyl sulfoxide solvent and other soluble impurities in the dispersion liquid; the normal saline is used for fixing the volume, and the sitagliptin nanometer preparation solution with the required concentration is obtained;
the rotational speed of centrifugal impurity removal in the step 6 is 14000 r/min, the centrifugal time is 15 min, the solution is placed in a dialysis bag with the cut-off molecular weight of 10kDa during dialysis, the Tris-HCl solution containing 0.9% NaCl is used as external liquid, the external liquid is dialyzed for 24-48 hours, and the external liquid is replaced for 3-5 times during dialysis.
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