CN111991348B - Doramectin slow-release microcapsule injection - Google Patents
Doramectin slow-release microcapsule injection Download PDFInfo
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Abstract
The invention belongs to the field of antiparasitic pharmaceutical preparations, and in particular relates to a doramectin slow-release microcapsule injection which comprises doramectin microcapsule particles, a wetting agent, a suspending agent, a deflocculant and water for injection; wherein, the doramectin microcapsule particles take silica aerogel particles loaded with doramectin as capsule cores and polylactic acid as capsule membranes. The polylactic acid coated silica aerogel-doramectin microcapsule particles prepared by the method have a slow release effect, are stable to light, are prepared into injection, and can achieve long-acting effect after being administrated in animal experiments.
Description
Technical Field
The invention belongs to the field of antiparasitic pharmaceutical preparations, and in particular relates to a doramectin slow-release microcapsule injection.
Background
Animal parasitic diseases are common diseases of animal husbandry in China, are various and widely distributed, and cause great harm to the animal husbandry. The doramectin is an avermectin antiparasitic drug which is prepared by taking cyclohexanecarboxylic acid as a precursor and fermenting a new strain of streptomyces avermitilis through gene recombination, is a 3 rd generation derivative of avermectin, and is considered as one of the most excellent antiparasitic drugs in the current avermectin family. Doramectin has high fat solubility, is easily dissolved in organic solvents such as methanol, ethanol, acetone, propylene glycol, dimethyl sulfoxide, ethyl acetate, isopropyl acetate and the like, has low solubility in water, and is sensitive to acid and light. Because doramectin has poor water solubility, the conventional dosage form is an oily injection, the phenomenon of peak valley appears in drug release in livestock and poultry, the poisoning of livestock and poultry is easy to cause, the half life period is short, the stability is poor, a large number of repeated dosing is needed, the labor cost is increased, and the repeated dosing of some dosage forms brings obvious stimulating reaction to the livestock and poultry, so that the healthy growth of the livestock and poultry is influenced.
Disclosure of Invention
The invention aims to overcome the defects and the shortcomings in the prior art and provide a doramectin slow-release microcapsule injection.
The technical scheme adopted by the invention is as follows: a doramectin sustained release microcapsule injection comprises doramectin microcapsule particles, a wetting agent, a suspending agent, a deflocculating agent and water for injection;
wherein, the doramectin microcapsule particles take silica aerogel particles loaded with doramectin as capsule cores and polylactic acid as capsule membranes.
The preparation process of the doramectin-loaded silica aerogel particles comprises the following steps:
(1-1) dissolving doramectin in a first organic solvent to prepare a saturated doramectin solution;
(1-2) dispersing silica aerogel powder in a saturated doramectin solution, stirring under a heating reflux, and removing a solvent to obtain silica aerogel-loaded doramectin drug-carrying particles;
the first organic solvent is one or more of chloroform, ethyl acetate, methanol, acetone and ethanol.
The preparation process of the silica aerogel powder comprises the following steps:
(a) Adding ethyl orthosilicate into deionized water, and slowly adding ethanol until the mixed solution becomes clear;
(b) Stirring, regulating pH to 3.0-4.0, standing at room temperature for 12-24 hr, and adding pore-enlarging agent;
(c) Mixing an emulsifying system by taking normal hexane as an oil phase, uniformly stirring, adding the solution added with the pore-enlarging agent in the step (b) into the oil phase under stirring, regulating the pH to 7.0-8.0 under stirring, continuously stirring, standing, forming wet gel, adding a protective solution, and putting into a closed container for aging;
(d) Adding n-hexane into the wet gel after aging, soaking, performing solvent exchange, and replacing n-hexane for multiple times;
(e) Removing the solvent to obtain silica aerogel powder;
(f) Roasting to remove the pore-expanding agent.
The process of forming the polylactic acid capsule outside the doramectin-loaded silica aerogel particles comprises the following steps:
(2-1) respectively dissolving poly-L-lactic acid and poly-D-lactic acid in a second organic solvent to obtain a poly-L-lactic acid solution and a poly-D-lactic acid solution, and respectively adding doramectin to saturation;
(2-2) adding the silica aerogel particles loaded with doramectin into the poly L-lactic acid solution or the poly D-lactic acid solution, uniformly dispersing the silica aerogel particles, vibrating, centrifuging and washing to obtain a layer of polylactic acid coated microcapsule particles;
(2-3) adding the once-coated microcapsule particles loaded into the poly-D-lactic acid solution or the poly-L-lactic acid solution, uniformly dispersing the once-coated microcapsule particles, vibrating, centrifuging and washing to obtain two layers of polylactic acid-coated microcapsule particles;
the polylactic acid in the step (2-2) and the polylactic acid in the step (2-3) are mutually isomeric.
The second organic solvent is one of chloroform, dichloromethane and acetone.
The wetting agent is one or more of ethanol, propanol, propylene glycol, glycerol, isopropanol, glycerol, tween-20, tween-40, tween-60, tween-80, tween-85 and polyethylene glycol.
The suspending agent is sodium carboxymethyl cellulose or methyl cellulose.
The deflocculating agent is sodium citrate.
The beneficial effects of the invention are as follows: the polylactic acid coated silica aerogel-doramectin microcapsule particles prepared by the method have a slow release effect, are stable to light, are prepared into injection, and can achieve long-acting effect after being administrated in animal experiments.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions of the prior art, the drawings which are required in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that it is within the scope of the invention to one skilled in the art to obtain other drawings from these drawings without inventive faculty.
FIG. 1 is a slow release profile of polylactic acid-coated silica aerogel-doramectin microcapsule particles and drug-loaded particles prepared newly in examples 1-3;
FIG. 2 is a graph showing the time course of the blood concentration in sheep for examples 4-6.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings, for the purpose of making the objects, technical solutions and advantages of the present invention more apparent.
1. Preparation of silica aerogel particles:
(a) Adding ethyl orthosilicate into deionized water, and slowly adding ethanol until the mixed solution becomes clear;
(b) Adding dilute hydrochloric acid under stirring to adjust the pH to 3.0-4.0, standing at room temperature for 12-24h, adding a pore-enlarging agent, and preparing a water phase system;
(c) Mixing and uniformly stirring an emulsion system by taking normal hexane as an oil phase to prepare an oil phase system, adding the water phase system in the step (b) into the oil phase system under stirring, adding ammonia water under stirring to adjust the pH to 7.0-8.0, continuously stirring for 15min, standing for forming wet gel, adding a protection liquid, and putting into a closed container for aging;
(d) Adding n-hexane into the wet gel after aging, soaking, performing solvent exchange, and replacing n-hexane for multiple times;
(e) Removing the solvent to obtain silica aerogel powder;
(f) Roasting to remove the pore-expanding agent.
Wherein, the emulsification system adopts as follows:
emulsification system 1: ethylene glycol fatty acid, tween 80=8:2;
emulsifying system 2: ethylene glycol fatty acid:tween 80=10:1;
emulsifying system 3: ethylene glycol fatty acid:tween 80, glycerol=8:2:0.1;
emulsifying system 4: glycol fatty acid, tween 80, glycerol=10:1:0.1.
Table 2 shows silica aerogel samples obtained under different preparation conditions.
2. Preparation of silica aerogel drug-loaded powder
The determination of the actual drug loading amount of the drug loading particles is carried out by a thermogravimetric analysis method, and the specific process is as follows: the temperature is heated from room temperature to 900 ℃ under the protection of nitrogen by a comprehensive thermal analysis instrument DTG60, and the heating rate is 10 ℃/min. According to the weight loss difference of the sample and the blank sample (namely the used silica aerogel sample) after drug loading, and subtracting the quantity of the blank sample proportionally, the actual drug loading quantity of the drug loading particles can be calculated.
1. Influence of the selection of different solvents on the actual drug loading:
considering that the solvent needs to be removed in the subsequent treatment process, a conventional low-boiling point solvent is adopted for the test, and the specific test process is as follows:
50g of sample 9 is divided into 5 groups, and is respectively added into 100mL of doramectin-chloroform saturated solution, 100mL of doramectin-ethyl acetate saturated solution, 100mL of doramectin-methanol saturated solution, 100mL of doramectin-propylene glycol saturated solution, 100mL of doramectin-acetone saturated solution and 100mL of doramectin-ethanol saturated solution, the sample 9 is uniformly distributed in the solution by ultrasonic oscillation, stirring is carried out for 2h under heating reflux, cooling is carried out to room temperature, filtering is carried out, filter cakes are washed by ethanol for multiple times, and then the filter cakes are dried to constant weight, thus obtaining the drug carrying particles 1-5. The actual drug loading of the drug-loaded particles 1-5 is 21.58%, 25.15%, 45.26%, 32.15% and 40.15% respectively.
The selection of different solvents can be seen to have a greater impact on the actual drug loading, wherein the alcoholic solvents facilitate drug adsorption into the silica aerogel.
2. Effect of silica aerogel pore size on actual drug loading
10g of sample 12, sample 13, sample 14 and sample 16 are taken and respectively added into 100mL of doramectin-methanol saturated solution, the samples are uniformly distributed in the solution by ultrasonic oscillation, stirring is carried out for 2 hours under heating reflux, cooling is carried out to room temperature, filtering is carried out, filter cakes are washed by ethanol for multiple times, and then the filter cakes are dried to constant weight, thus obtaining the drug-loaded particles 6-9. The actual drug loading of the drug-loaded particles 6-9 was measured to be 34.26%, 52.26%, 47.64% and 56.21% respectively.
Comparing the drug-loaded particles 3 prepared under the same condition, the larger the pore diameter of the silica aerogel is, the larger the actual drug-loaded amount is.
3. Influence of particle size of silica aerogel on actual drug loading
Taking 10g of sample 17 and sample 18, respectively adding into 100mL of doramectin-methanol saturated solution, vibrating with ultrasonic waves to uniformly distribute the sample in the solution, stirring for 2 hours under heating and refluxing, cooling to room temperature, filtering, washing a filter cake with ethanol for multiple times, and then drying to constant weight to obtain the drug-loaded particles 10-11. The actual drug loading of the drug-loaded particles 10-11 is 57.14% and 56.98% respectively.
Compared with the drug-loaded particles 16 prepared under the same conditions, the particle size of the silica aerogel has less influence on the actual drug loading.
The particle size of the silica aerogel does not change much before and after drug loading.
3. Preparation of polylactic acid coated silica aerogel-doramectin microcapsule particles
Example 1:
(1) Respectively dissolving poly L-lactic acid and poly D-lactic acid with molecular weight of 1-10W in chloroform to obtain poly L-lactic acid solution with the molecular weight of 10g/L and poly D-lactic acid solution with the molecular weight of 10g/L, respectively adding excessive doramectin, stirring, and filtering to obtain poly L-lactic acid solution and poly D-lactic acid solution containing saturated doramectin.
(2) Adding the drug-loaded particles into a 16 centrifuge tube, adding the drug-loaded particles into a poly L-lactic acid solution, vibrating to disperse the particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(3) Adding a poly-D-lactic acid solution into a centrifuge tube, vibrating to disperse particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(4) And freeze-drying to constant weight to obtain the polylactic acid coated silica aerogel-doramectin microcapsule particles.
Compared with the drug-loaded particles 16, the polylactic acid coated silica aerogel-doramectin microcapsule particles prepared in the embodiment have the weight gain of 14.27%, the theoretical drug-loaded rate of 49.19% and the actual drug-loaded rate of 50.62%. The average particle size was 23.24nm and the particle size distribution was as follows: 0-10nm,5%;10-20nm,27%;20-30nm,38%; 30-40nm,21%;40-50nm,7%;50-60,2%.
Example 2:
(1) Respectively dissolving poly L-lactic acid and poly D-lactic acid with molecular weight of 1-10W in chloroform to obtain poly L-lactic acid solution with the molecular weight of 10g/L and poly D-lactic acid solution with the molecular weight of 10g/L, respectively adding excessive doramectin, stirring, and filtering to obtain poly L-lactic acid solution and poly D-lactic acid solution containing saturated doramectin.
(2) Adding the drug-loaded particles 17 into a centrifuge tube, adding the drug-loaded particles into a poly L-lactic acid solution, vibrating to disperse the particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(3) Adding a poly-D-lactic acid solution into a centrifuge tube, vibrating to disperse particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(4) And freeze-drying to constant weight to obtain the polylactic acid coated silica aerogel-doramectin microcapsule particles.
Compared with the drug-loaded particles 17, the polylactic acid coated silica aerogel-doramectin microcapsule particles prepared in the embodiment have the weight gain of 10.75%, the theoretical drug-loaded rate of 49.19% and the actual drug-loaded rate of 50.62%. The average particle size was 55.39nm and the particle size distribution was as follows: 20-30nm,6%;30-40nm,8%;40-50nm, 22%;50-60, 31%;60-70nm,18%;70-80nm,9%;80-90nm,5%.
Example 3:
(1) Respectively dissolving poly-L-lactic acid and poly-D-lactic acid with molecular weight of 1-10W in chloroform to obtain poly-L-lactic acid solution with the concentration of 10g/L and poly-D-lactic acid solution with the concentration of 10g/L, respectively adding excessive doramectin, stirring, and filtering to obtain poly-L-lactic acid solution and poly-D-lactic acid solution containing saturated doramectin.
(2) Adding the drug-loaded particles 18 into a centrifuge tube, adding the drug-loaded particles into a poly L-lactic acid solution, vibrating to disperse the particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(3) Adding a poly-D-lactic acid solution into a centrifuge tube, vibrating to disperse particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(4) And freeze-drying to constant weight to obtain the polylactic acid coated silica aerogel-doramectin microcapsule particles.
Compared with the drug-loaded particles 18, the polylactic acid coated silica aerogel-doramectin microcapsule particles prepared in the embodiment have the weight gain of 8.18%, the theoretical drug-loaded rate is 52.67%, and the actual drug-loaded rate is 51.62%. The average particle size was 83.6nm and the particle size distribution was as follows: 50-60,1%;60-70nm,7%;70-80nm,26%; 80-90nm,35%;90-100nm,19%;100-110nm,10%;110-120nm,2%.
Comparative example 1:
(1) Poly (L-lactic acid) and poly (D-lactic acid) having molecular weights of 1 to 10W were dissolved in chloroform, respectively, to obtain a poly (L-lactic acid) solution of 10g/L and a poly (D-lactic acid) solution of 10 g/L.
(2) Adding the drug-loaded particles into a 16 centrifuge tube, adding the drug-loaded particles into a poly L-lactic acid solution, vibrating to disperse the particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(3) Adding a poly-D-lactic acid solution into a centrifuge tube, vibrating to disperse particles, vibrating for 1h at 40 ℃, and centrifugally washing for 3 times by using chloroform, wherein the centrifugal parameter is 1000r/min and 1min;
(4) And freeze-drying to constant weight to obtain the polylactic acid coated silica aerogel-doramectin microcapsule particles, wherein the weight of the polylactic acid coated silica aerogel-doramectin microcapsule particles is reduced by 23.89 percent compared with the drug-loaded particles 16.
The supernatants centrifuged in step (2) and step (3) of comparative example 1 were examined to detect doramectin, and it was found that doramectin was eluted from silica aerogel particles loaded with doramectin during the formation of the polylactic acid capsule film, and the total amount of elution was calculated to be 46.94% of the total amount of actual drug loading of the drug-loaded particles 16.
1. Establishment of doramectin standard curve
Accurately weighing a proper amount of doramectin standard substance, adding 50% ethanol solution, diluting to prepare 0.5mg/mL solution, and scanning the doramectin solution by an ultraviolet-visible absorption spectrometer to find the maximum absorption wavelength of doramectin.
0.005, 0.01, 0.015, 0.02, 0.03 and 0.05mg/mL of doramectin solutions with 5 different concentrations are prepared by using 50% ethanol solution, the absorbance at the maximum absorption wavelength is measured, the absorbance is measured by taking ethyl acetate as a blank control and respectively measuring the absorbance at the maximum absorption wavelength, and the linear regression analysis is carried out on the absorbance and the mass concentration to obtain a doramectin standard curve.
2. Theoretical drug loading and actual drug loading
The theoretical drug loading is calculated by assuming that the amount of doramectin in the drug loading particles is unchanged and the weight increasing part is polylactic acid.
The detection method of the actual drug loading is as follows: sample 0.1g was taken and placed in a test tube, digested with protease solution for 24 hours, after complete digestion, centrifuged and washed, and the liquid fractions were pooled. Adding 3mL of ethyl acetate into the liquid part, carrying out vortex oscillation for 15min, standing and layering, taking the upper ethyl acetate layer, repeating for 3 times, combining ethyl acetate extracting solutions, filtering through a 0.45 mu m filter membrane, adding into a 15mL volumetric flask, and fixing the volume by using ethyl acetate. And (3) under the detection wavelength, measuring an absorbance value by taking ethyl acetate as a reference liquid, and calculating the doramectin content in the liquid part. And detecting the doramectin content in the filter residue by adopting a thermogravimetric analysis method in the solid part. The sum of the doramectin contents of the two parts is the actual drug loading of doramectin in 0.1g of the sample.
3. Establishment of doramectin slow release curve
Accurately weighing a certain amount of doramectin (control group), drug-loaded particles 16 and a sample, respectively placing the doramectin, the drug-loaded particles 16 and the sample in a cylindrical dialysis bag, clamping two ends by dialysis clamps, suspending the doramectin, the drug-loaded particles and the sample in a three-hole beaker containing a certain amount of buffer solution (50% ethanol solution), sealing the doramectin, placing the doramectin and the sample in a bath, performing constant-temperature water bath vibration (the temperature is 37.5 ℃) in a dark state, taking a certain amount of buffer solution outside the dialysis bag at different time points, and simultaneously supplementing the same volume of fresh buffer solution. The absorbance values were measured to obtain doramectin concentrations in the different time solutions and the cumulative release fractions were calculated.
FIG. 1 is a graph showing the slow release profile of polylactic acid-coated silica aerogel-doramectin microcapsule particles and drug-loaded particles 16-18 prepared in examples 1-3, wherein the release of doramectin was slower after the polylactic acid coating.
The silica aerogel-doramectin microcapsule particles and drug-loaded particles 16 prepared in example 1 were each enclosed in an ampoule bottle, and subjected to a sustained release experiment by (4500±500) LX intense light irradiation for 10 d. The cumulative release amounts of the buffer solution taken in the slow release 10d were 29.86% and 45.69%, respectively, and the peak areas of the isomerized compound and the non-isomerized compound in the reaction solution were measured by HPLC, and the isomerized compound contents were calculated to be 1.05% and 25.65%, respectively.
The silica aerogel-doramectin microcapsule particles prepared in example 2 were sealed in ampoule bottles, and subjected to a sustained release experiment under (4500±500) LX intense light irradiation for 10 d. The cumulative release amounts of the buffer solution taken in the slow release 10d were 27.98% and 48.25%, respectively, and the peak areas of the isomerized compound and the non-isomerized compound in the reaction solution were measured by HPLC, and the isomerized compound contents were calculated to be 0.96% and 29.25%, respectively.
The silica aerogel-doramectin microcapsule particles prepared in example 3 were sealed in ampoule bottles, and subjected to a sustained release experiment under (4500.+ -. 500) LX intense light irradiation for 10 d. The cumulative release amounts of the buffer solution taken in the slow release 10d were 32.86% and 49.26%, respectively, and the peak areas of the isomerized compound and the non-isomerized compound in the reaction solution were measured by HPLC, and the isomerized compound contents were calculated to be 1.41% and 36.15%, respectively.
4. Preparation of doramectin slow-release microcapsule injection and slow-release research of doramectin slow-release microcapsule injection in animal bodies
The examples 1-3 were tested with different wetting agents, suspending agents, deflocculants and amounts, respectively, to obtain a relatively stable preferred example of suspension system, as follows:
example 4:
a doramectin slow-release microcapsule injection is prepared by the following steps:
(1) Raw material preparation: 5.0% of the silica aerogel-doramectin microcapsule particles prepared in the example 1, 2.6% of a wetting agent polyethylene glycol-200, 1.5% of a suspending agent sodium hydroxymethyl cellulose, 1.3% of a deflocculant sodium citrate and the balance of water for injection.
(2) Adding the wetting agent into 2/3 of water for injection, and stirring to uniformly mix;
(3) Dissolving a suspending agent and a deflocculating agent in the rest water for injection;
(4) And (3) shearing and dispersing the drug-carrying particles into the solution obtained in the step (2), shearing for 20 minutes, adding the solution obtained in the step (3), and shearing for 20 minutes to obtain the doramectin slow-release microsphere injection.
Example 5:
a doramectin slow-release microcapsule injection is prepared by the following steps:
(1) Raw material preparation: 5.0% of the silica aerogel-doramectin microcapsule particles prepared in the example 2, 3.1% of a wetting agent polyethylene glycol-200, 1.5% of a suspending agent sodium hydroxymethyl cellulose, 1.5% of a deflocculant sodium citrate and the balance of water for injection.
(2) Adding the wetting agent into 2/3 of water for injection, and stirring to uniformly mix;
(3) Dissolving a suspending agent and a deflocculating agent in the rest water for injection;
(4) And (3) shearing and dispersing the drug-carrying particles into the solution obtained in the step (2), shearing for 20 minutes, adding the solution obtained in the step (3), and shearing for 20 minutes to obtain the doramectin slow-release microsphere injection.
Example 6:
a doramectin slow-release microcapsule injection is prepared by the following steps:
(1) Raw material preparation: 5.0% of the silica aerogel-doramectin microcapsule particles prepared in the example 3, 3.5% of a wetting agent polyethylene glycol-400, 1.5% of a suspending agent sodium hydroxymethyl cellulose, 1.5% of a deflocculant sodium citrate and the balance of water for injection.
(2) Adding the wetting agent into 2/3 of water for injection, and stirring to uniformly mix;
(3) Dissolving a suspending agent and a deflocculating agent in the rest water for injection;
(4) And (3) shearing and dispersing the drug-carrying particles into the solution obtained in the step (2), shearing for 20 minutes, adding the solution obtained in the step (3), and shearing for 20 minutes to obtain the doramectin slow-release microsphere injection.
Wherein, the 3 hour sedimentation volume ratio of the embodiment 4 to the embodiment 6 is 96.5 percent, 96 percent and 94.5 percent respectively, which meets the requirement that the 90 percent sedimentation volume of the suspension is 3 hours specified by pharmacopoeia, and the embodiment 18 to the embodiment 20 are evenly mixed again and need to be turned upside down for 4 times (easy dispersion), 6 times (easy dispersion) and 6 times (easy dispersion) respectively.
Pharmacokinetic assay method: 5 sheep are injected with a doramectin original drug suspension (comparison group) and a doramectin slow-release microcapsule injection by subcutaneous injection at the neck of 20mg/kg, 10mL of blood is taken by vein after administration of 10, 20, 30, 60, 120 and 180d respectively, heparin is anticoagulated, centrifugation is carried out at 4000r/min for 20min, 2.0mL of plasma sample is precisely sucked, 20.0 mu L of internal standard solution is added, vortex mixing is carried out for 2min, 2.0mL of ethyl acetate is added, vortex mixing is carried out for 2min, centrifugation is carried out for 10min, the rotating speed is 12000r/min, the upper organic phase is taken, the same operation is carried out for 3 times (the ethyl acetate dosage is 2.0,1.5 and 1.5mL respectively), the upper liquid is combined, water bath at 50 ℃ is carried out, nitrogen blow-drying is carried out, 100.0 mu L of methanol is added, vortex is carried out for 2min, centrifugation is carried out for 10min at 12000r/min, supernatant liquid is taken and passes through a microporous filter membrane of 0.22 mu m, the doramectin content is detected by high performance liquid chromatography, and the average blood mass concentration is calculated.
As shown in fig. 2, the embodiment of the present invention can achieve slow release and maintain long-acting in sheep.
The foregoing disclosure is illustrative of the present invention and is not to be construed as limiting the scope of the invention, which is defined by the appended claims.
Claims (5)
1. A doramectin slow-release microcapsule injection is characterized in that: comprises doramectin microcapsule particles, a wetting agent, a suspending agent, a deflocculating agent and water for injection;
wherein, the doramectin microcapsule particles take silica aerogel particles loaded with doramectin as capsule cores and polylactic acid as capsule membranes;
the preparation process of the silica aerogel powder comprises the following steps:
(a) Adding ethyl orthosilicate into deionized water, and slowly adding ethanol until the mixed solution becomes clear;
(b) Stirring, regulating pH to 3.0-4.0, standing at room temperature for 12-24 hr, and adding pore-enlarging agent;
(c) Mixing an emulsifying system by taking normal hexane as an oil phase, uniformly stirring, adding the solution added with the pore-enlarging agent in the step (b) into the oil phase under stirring, regulating the pH to 7.0-8.0 under stirring, continuously stirring, standing, forming wet gel, adding a protective solution, and putting into a closed container for aging;
(d) Adding n-hexane into the wet gel after aging, soaking, performing solvent exchange, and replacing n-hexane for multiple times;
(e) Removing the solvent to obtain silica aerogel powder;
(f) Roasting to remove the pore-expanding agent;
wherein the volume ratio of the water to the oil is 1:3 or 2:1, and the emulsifying system is as follows: ethylene glycol fatty acid:tween 80:glycerin=8:2:0.1, the adding amount of an emulsifying system is 0.5 or 0.9g/mL, the pore-expanding agent is urotropine, and the adding amount of the pore-expanding agent is 0.07 or 0.09g/mL;
the preparation process of the doramectin-loaded silica aerogel particles comprises the following steps:
(1-1) dissolving doramectin in a first organic solvent to prepare a saturated doramectin solution;
(1-2) dispersing silica aerogel powder in a saturated doramectin solution, stirring under a heating reflux, and removing a solvent to obtain silica aerogel-loaded doramectin drug-carrying particles;
the first organic solvent is one or more of chloroform, ethyl acetate, methanol, acetone and ethanol;
the process of forming the polylactic acid capsule outside the doramectin-loaded silica aerogel particles comprises the following steps:
(2-1) respectively dissolving poly-L-lactic acid and poly-D-lactic acid in a second organic solvent to obtain a poly-L-lactic acid solution and a poly-D-lactic acid solution, and respectively adding doramectin to saturation;
(2-2) adding the silica aerogel particles loaded with doramectin into the poly L-lactic acid solution or the poly D-lactic acid solution, uniformly dispersing the silica aerogel particles, vibrating, centrifuging and washing to obtain a layer of polylactic acid coated microcapsule particles;
(2-3) adding the once-coated microcapsule particles loaded into the poly-D-lactic acid solution or the poly-L-lactic acid solution, uniformly dispersing the once-coated microcapsule particles, vibrating, centrifuging and washing to obtain two layers of polylactic acid-coated microcapsule particles;
the polylactic acid in the step (2-2) and the polylactic acid in the step (2-3) are mutually isomeric.
2. The doramectin sustained-release microcapsule injection according to claim 1, characterized in that: the second organic solvent is one of chloroform, dichloromethane and acetone.
3. The doramectin sustained-release microcapsule injection according to claim 1, characterized in that: the wetting agent is one or more of ethanol, propanol, propylene glycol, glycerol, isopropanol, glycerol, tween-20, tween-40, tween-60, tween-80, tween-85 and polyethylene glycol.
4. The doramectin sustained-release microcapsule injection according to claim 1, characterized in that: the suspending agent is sodium carboxymethyl cellulose or methyl cellulose.
5. The doramectin sustained-release microcapsule injection according to claim 1, characterized in that: the deflocculating agent is sodium citrate.
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| WO2014090173A1 (en) * | 2012-12-13 | 2014-06-19 | 清华大学深圳研究生院 | Nanoscale doxorubicin hydrochloride and preparation method therefor |
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