CN111643451B - Honokiol self-emulsifying microemulsion preparation for injection and preparation method thereof - Google Patents
Honokiol self-emulsifying microemulsion preparation for injection and preparation method thereof Download PDFInfo
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Abstract
The invention provides a honokiol self-emulsifying nano microemulsion preparation for injection and a preparation method thereof, belonging to the technical field of medicine preparation, wherein the honokiol self-emulsifying microemulsion preparation comprises the following raw materials in parts by weight: 40-60 parts of honokiol, 200-250 parts of oil phase, 400-440 parts of emulsifier, 290-330 parts of co-emulsifier and 4-8 parts of antioxidant; the oil phase comprises an injectable grade medium chain triglyceride; the emulsifier comprises polyethylene glycol 12 hydroxystearic acid 15 and egg yolk lecithin; the coemulsifier comprises absolute ethyl alcohol for injection; the antioxidant comprises acetylcysteine; the honokiol self-emulsifying microemulsion preparation for injection has good self-emulsifying effect, good particle size and PDI of the formed nano emulsion and good stability; the injection honokiol self-emulsifying microemulsion preparation has the effects of prolonging the blood coagulation time of mice and increasing the whole cerebral ischemia of the mice.
Description
Technical Field
The invention belongs to the technical field of medicine preparation, and particularly relates to a honokiol self-emulsifying microemulsion preparation for injection and a preparation method thereof.
Background
The honokiol is magnolol isomer, and is effective component of cortex Magnolia officinalis. Is a dimer polymerized by the side chain of monopropyrin and the benzene nucleus of another phenylpropanoid, and has the following structural formula:
the honokiol is crystalline powder, has molecular weight of 266.33, melting point of 87.5 deg.C, and dissociation constant pKa1＝9.64±0.30，pKa2＝10.71±0.21。
In the pharmacodynamic research process, the honokiol not only has the antibacterial, anti-inflammatory and anti-ulcer effects related to the traditional function and main treatment effect, but also has the effects of resisting oxidation, myocardial ischemia, arrhythmia and the like; and the honokiol can penetrate the blood brain barrier and has high content in brain tissue, which indicates that the honokiol can play a role in the treatment of ischemic cerebral apoplexy.
However, honokiol is poorly absorbed orally, the bioavailability of oral absorption is less than 10%, and a large amount of drugs are retained in intestinal tracts, which seriously damage intestinal flora and easily cause intestinal flora imbalance. In view of the poor water solubility of honokiol, the saturated molarity of honokiol is only 27. mu. mol.L when dissolved in HBSS buffer solution (pH 7.4)-1I.e., 7.19. mu.g/mL, is difficult to prepare into an aqueous injection solution.
The microemulsion is a novel drug carrier system, and is a transparent, semitransparent, low-viscosity, isotropic and thermodynamically stable oil-water mixing system spontaneously formed by a water phase, an oil phase, a surfactant and a cosurfactant in a proper proportion. The particle size of microemulsion droplets is usually below hundreds of nanometers, the microemulsion droplets can obviously improve the solubility of insoluble drugs when being used as a drug carrier, have better stability compared with the common emulsion, can increase the in vivo bioavailability of active substances due to high dispersion and stronger tissue affinity, and have wide application prospect when being used as a drug carrier.
Self-emulsifying microemulsion (Self-emulsifying microemulsion) is a new preparation technology developed on the basis of the excellent performance of microemulsion, and the Self-emulsifying microemulsion is an oily mixture composed of medicine, oil, emulsifier, cosolvent and the like, and can spontaneously form a thermodynamically stable, uniform, transparent or semitransparent isotropic solution under the slight in vitro oscillation or the in vivo gastrointestinal tract peristalsis. The oil solution of the self-emulsifying microemulsion preparation is prepared by simply mixing a proper emulsifier, a co-emulsifier, an oil phase and raw material medicines, is easy to store, and is stable and uniform. When in use, the oil solution and the diluent are mixed, and the microemulsion injection with clear and transparent property and slight blue opalescence can be formed after slight shaking, thereby greatly improving the stability of the raw material medicaments in the preparation and storage processes, obviously increasing the solubility and bioavailability of the medicaments and improving the pharmacological effect of the medicaments.
At present, no self-emulsifying microemulsion preparation of honokiol exists in the market.
Disclosure of Invention
In view of the above, the present invention aims to provide a honokiol self-emulsifying microemulsion preparation for injection and a preparation method thereof; the honokiol self-emulsifying microemulsion preparation has good self-emulsifying effect, good particle size and PDI of the formed nano emulsion and good stability; the honokiol self-emulsifying microemulsion preparation has the effects of prolonging the blood coagulation time of mice and increasing the whole cerebral ischemia of the mice.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a honokiol self-emulsifying microemulsion preparation for injection, which comprises the following raw materials in parts by weight: 40-60 parts of honokiol, 200-250 parts of oil phase, 400-440 parts of emulsifier, 290-330 parts of co-emulsifier and 4-8 parts of antioxidant;
preferably, the feed comprises the following raw materials in parts by weight: 45-55 parts of honokiol, 210-230 parts of oil phase, 410-430 parts of emulsifier, 300-310 parts of co-emulsifier and 5-7 parts of antioxidant.
Preferably, the emulsifier comprises one or a mixture of more of injection-grade polyethylene glycol 12 hydroxystearic acid 15, tween 80, span 80, polyoxyethylene ether 40 hydrogenated castor oil, polyoxyethylene ether 35 hydrogenated castor oil, pluronic F68, poloxamer 188, lecithin and soybean lecithin;
preferably one or a mixture of more of polyethylene glycol 12 hydroxystearic acid 15, tween 80, polyoxyethylene ether 40 hydrogenated castor oil, pluronic F68 and lecithin;
preferably one or more of polyethylene glycol 12 hydroxystearic acid 15, polyoxyethylene ether 40 hydrogenated castor oil, polyoxyethylene ether 35 hydrogenated castor oil, pluronic F68 and lecithin;
polyethylene glycol 12 hydroxystearic acid 15 and egg yolk lecithin are more preferable.
The oil phase comprises one or more of injection-grade medium-chain triglyceride, ethyl acrylate, isopropyl palmitate and isopropyl myristate;
preferably medium chain triglycerides or isopropyl myristate;
still more preferred are medium chain triglycerides.
The auxiliary emulsifier comprises one or a mixture of more of anhydrous ethanol for injection, polyethylene glycol 400, polyethylene glycol 200, glycerol and 1, 2-propylene glycol.
Preferably one or a mixture of several of absolute ethyl alcohol, polyethylene glycol 400 and 1, 2-propylene glycol;
anhydrous ethanol is more preferred.
The antioxidant comprises one or more of sodium sulfite, sodium pyrosulfite, dibutyl phenol, sodium bisulfite, sodium thiosulfate, tert-butyl p-hydroxyanisole, thiourea, vitamin C, propyl gallate, alpha-tocopherol, ascorbyl palmitate, and acetylcysteine;
preferably one or a mixture of more of sodium metabisulfite, dibutyl phenol, vitamin C, alpha-tocopherol, ascorbyl palmitate and acetylcysteine;
more preferably acetylcysteine.
Preferably, the weight part of the egg yolk lecithin is 5-55 parts, and the weight part of the polyethylene glycol 12 hydroxystearic acid 15 is 385-395 parts.
The invention also provides a preparation method of the honokiol self-emulsifying microemulsion preparation for injection, which comprises the following steps:
1) mixing egg yolk lecithin, an antioxidant and an auxiliary emulsifier, and then mixing the mixture with polyethylene glycol 12 hydroxystearic acid 15 and an oil phase to obtain a solvent solution;
2) mixing honokiol with the solvent solution to obtain the honokiol self-emulsifying microemulsion preparation.
9. The method of claim 8, wherein the mixing of the honokiol and the solvent solution in step 2) further comprises filtration with a filter membrane; the pore size of the filter membrane is 0.22 μm.
The invention also provides an injection preparation comprising the self-emulsifying microemulsion preparation for injection, and also comprises 5% glucose injection or 0.9% NaCl injection.
The invention has the beneficial effects that: the honokiol self-emulsifying microemulsion preparation provided by the invention comprises the following raw materials in parts by weight: 40-60 parts of honokiol, 200-250 parts of oil phase, 400-440 parts of emulsifier, 290-330 parts of co-emulsifier and 4-8 parts of antioxidant; the emulsifier comprises polyethylene glycol 12 hydroxystearic acid 15 and egg yolk lecithin. In the invention, the concentration of honokiol in the self-emulsifying microemulsion preparation can reach 50mg/g, and the solubility of the active ingredient honokiol is greatly improved; has better stability, and can increase the in vivo bioavailability of active substance honokiol and avoid the influence of the medicine on normal flora of intestinal tract due to high dispersion and strong tissue affinity.
The honokiol self-emulsifying microemulsion preparation provided by the invention is used for treating mice with the dosage of 70 mu g/kg, and has the effects of prolonging the blood coagulation time of the mice and increasing the whole cerebral ischemia of the mice. The blood coagulation time shows that the pharmacodynamic result of the honokiol self-emulsifying microemulsion preparation provided by the invention is slightly superior to that of butylphthalide and edaravone. The whole brain ischemia test shows that the four patients have no obvious difference, but the honokiol self-emulsifying microemulsion preparation provided by the invention has smaller dosage which is about 1 percent of butylphthalide and edaravone.
In clinical use, the honokiol self-emulsifying microemulsion preparation provided by the invention can adjust different administration concentrations according to the administration requirements of patients, and stable and completely clear nano emulsion can be rapidly formed after different dilution multiples of 5% glucose injection and 0.9% NaCl injection are adopted for dilution; the emulsion forming effect, the appearance of the emulsion and the size and the distribution of the grain diameter are not obviously changed. The nanoemulsion diluent is placed at room temperature for 1 month, and no obvious abnormality is found, which indicates that the microemulsion injection and the microemulsion prepared by diluting the microemulsion injection into an emulsion have good stability.
Detailed Description
The invention provides a honokiol self-emulsifying microemulsion preparation for injection, which comprises the following raw materials in parts by weight: 40-60 parts of honokiol, 200-250 parts of oil phase, 400-440 parts of emulsifier, 290-330 parts of co-emulsifier and 4-8 parts of antioxidant.
In the invention, the honokiol self-emulsifying microemulsion preparation comprises 40-60 parts of honokiol, preferably 45-55 parts of honokiol, and more preferably 50 parts of honokiol; the concentration of the honokiol carried in the self-emulsifying microemulsion preparation is preferably more than or equal to 50 mg/g.
In the invention, the honokiol is commercially or self-made honokiol; preferably, self-made honokiol with the purity of more than 99 percent is adopted; the preparation method comprises the following steps: 1) mixing cortex Magnolia officinalis with ethanol, extracting, and concentrating to obtain first cortex Magnolia officinalis extract; 2) mixing the first magnolia extract with calcium oxide or calcium hydroxide, standing, drying and crushing to obtain first magnolia extract powder; 3) mixing the first magnolia officinalis extract powder with ethanol for second extraction to obtain a second magnolia officinalis extract; 4) mixing the second magnolia officinalis extract with dichloromethane, and extracting with a sodium hydroxide solution to obtain an alkaline extract; 6) adjusting the alkaline extraction liquid to be neutral to obtain a precipitate; 7) dissolving the precipitate with petroleum ether, and crystallizing to obtain crude crystals of honokiol; 8) and dissolving the crude honokiol crystal with petroleum ether, and recrystallizing to obtain a pure honokiol product.
In the invention, magnolia officinalis is mixed with ethanol for first extraction and concentration to obtain a first magnolia officinalis extract. In the present invention, it is preferable that the pulverized magnolia officinalis is mixed with ethanol for the first extraction; the volume concentration of the ethanol is preferably 85-95%, and more preferably 90%; the volume of the ethanol is preferably 5-9 times of that of the magnolia officinalis. In the invention, the number of times of the first extraction is preferably 2-5 times, and more preferably 3 times; in the present invention, when the number of times of the first extraction is preferably 3 times; the volume ratio of the ethanol is preferably 8 times, 6 times and 6 times of the volume of the magnolia officinalis respectively; in the invention, the time for each extraction is preferably 1-3 h, and more preferably 1.5-2.5 h. After several times of extraction, combining extracting solutions; concentrating the extracting solution to obtain a first magnolia officinalis extract; the method for concentrating is not particularly limited, and the conventional concentrating method in the field is adopted; in the practice of the present invention, the concentration is preferably a concentration under reduced pressure.
After the first magnolia officinalis extract is obtained, the first magnolia officinalis extract is mixed with calcium oxide, and the mixture is stood, dried and crushed to obtain first magnolia officinalis powder. In the present invention, the mass of the calcium oxide is preferably 8% to 32%, preferably 9% to 30%, and more preferably 10% of the mass of the first magnolia bark extract. In the present invention, the calcium oxide is preferably finely powdered calcium oxide; before the first magnolia extract is mixed with calcium oxide, the first magnolia extract is preferably mixed with absolute ethyl alcohol, and the mass ratio of the volume of the absolute ethyl alcohol to the first magnolia extract is preferably (0.5-1) mL:1 g; in the invention, the anhydrous ethanol is used for dissolving the first magnolia extract, so that the first magnolia extract can be better mixed and reacted with calcium oxide. In the invention, the first magnolia extract and the calcium oxide are preferably stirred concomitantly in the mixing process, and the stirring speed of the invention has no special requirement, so that the first magnolia extract and the calcium oxide can be uniformly mixed. In the invention, the first magnolia extract is mixed with calcium oxide and then stands; the standing time is preferably 20-28 h, and more preferably 22-26 h. After the standing, the invention is dried and crushed; the method for drying and pulverizing is not particularly limited in the present invention, and a drying and pulverizing method which is conventional in the art may be employed. In the invention, the calcium oxide has the function of effectively removing impurities in the first magnolia bark extract; the calcium oxide is combined with the resin components and the oily substances in the first magnolia extract and is saponified to form insoluble substances, so that the impurity content is effectively reduced; meanwhile, as magnolol and honokiol are both acidic, the honokiol and calcium oxide can perform neutralization reaction; the calcium oxide firstly reacts with magnolol, and the quality of the calcium oxide is controlled within the range, so that the magnolol in the first magnolia officinalis extract can be effectively removed, and the honokiol is reserved.
In the invention, the first magnolia bark powder is mixed with ethanol for secondary extraction to obtain a second magnolia bark extract. In the present invention, the volume concentration of ethanol is preferably 85% to 95%, more preferably 90%; the volume of the ethanol is preferably 3-8 times, and more preferably 4-6 times of that of the first magnolia bark powder. In the invention, the number of times of the second extraction is preferably 2-5 times, and more preferably 3 times; in the invention, the time for each extraction is preferably 2-5 h, and more preferably 2.5-4 h. In the present invention, the ethanol extraction is preferably carried out in an extraction tank; after several times of extraction, combining extracting solutions; concentrating the extracting solution to obtain a second extract; the method for concentrating is not particularly limited, and the conventional concentrating method in the field is adopted; in the practice of the present invention, the concentration is preferably a concentration under reduced pressure.
After the second magnolia officinalis extract is obtained, the second magnolia officinalis extract is mixed with dichloromethane and then extracted by a sodium hydroxide solution to obtain an alkaline extraction liquid. In the invention, the mass ratio of the second magnolia extract to dichloromethane is preferably 1 (4-7), and more preferably 1 (4.5-6). In the invention, the second magnolia extract and dichloromethane are mixed and then preferably filtered, and in the invention, the filtering is preferably gauze filtering or filter paper filtering; after the filtrate is obtained, the filtrate is extracted by sodium hydroxide solution to obtain alkaline extract. In the invention, the number of extraction times of the sodium hydroxide solution is preferably 2-4, and more preferably 3; in the present invention, the mass concentration of the sodium hydroxide solution is preferably 0.8% to 5%, more preferably 1.0% to 3.0%; in the present invention, when the number of extractions is preferably 3; the volume ratio of the sodium hydroxide solution to the filtrate is preferably 1:2, 1:1 or 1: 1; the extraction is preferably carried out in a separatory funnel; after the extraction is finished, the alkaline extraction liquid is collected.
After the alkaline extraction liquid is obtained, the alkaline extraction liquid is adjusted to be neutral to obtain a precipitate. In the invention, the adjustment is preferably carried out by using brine, and the neutral preferable pH value is 7.0; the hydrochloric acid is not particularly required, and the hydrochloric acid solution which is conventional in the field is adopted. According to the invention, after the alkaline extraction liquid is adjusted to be neutral, a large amount of precipitate can appear.
After the precipitate is obtained, petroleum ether is used for dissolving the precipitate and then crystallizing to obtain crude crystals of honokiol. In the invention, the precipitate and petroleum ether are mixed and heated and refluxed until being dissolved; in the present invention, the temperature and time of the heating reflux are not particularly limited, and it is preferable to precipitate the whole solvent. In the invention, the boiling range of the petroleum ether is preferably 60-90 ℃; the mass ratio of the volume of the petroleum ether to the precipitate is preferably (20-30) mL:1 g; more preferably (22 to 27) mL:1 g. After the precipitate is completely dissolved, cooling to obtain crude crystals of honokiol; the cooling method is not particularly limited, and natural cooling is preferably performed; in the present invention, after the cooling, a vacuum drying step is preferably further included, and the parameters of the vacuum drying are not particularly limited in the present invention, and may be vacuum drying parameters conventional in the art.
After the crude honokiol crystal is obtained, the crude honokiol crystal is dissolved by petroleum ether and then recrystallized to obtain a pure honokiol product. In the invention, the boiling range of the petroleum ether is preferably 60-90 ℃; the mass ratio of the volume of the petroleum ether to the precipitate is preferably (25-40) mL:1 g; more preferably (28 to 35) mL:1 g. In the invention, the crude crystals of honokiol and petroleum ether are mixed, heated and refluxed until being dissolved; the temperature and time for heating and refluxing are not particularly limited, and all solvents for the magnolol group crystals are preferably used; when the crude honokiol crystals are completely dissolved, a reddish brown oily body appears at the lower layer; filtering to remove the red brown oil, cooling and recrystallizing to obtain a pure honokiol product; after the cooling recrystallization, the invention preferably further comprises a vacuum drying step, and the parameters of the vacuum drying are not particularly limited, and the vacuum drying parameters conventional in the field can be adopted. In the invention, the pure honokiol product is needle-shaped crystal.
In the invention, the honokiol self-emulsifying microemulsion preparation comprises 200-250 parts of oil phase, preferably 210-230 parts, and more preferably 218 parts; in the present invention, the oil phase is preferably a medium chain triglyceride; the source of the medium-chain triglycerides is not particularly limited in the present invention, and any commercially available product that is conventional in the art may be used.
In the invention, the honokiol self-emulsifying microemulsion preparation comprises 400-440 parts of an emulsifier; the emulsifier preferably comprises polyethylene glycol 12 hydroxystearic acid 15 and egg yolk lecithin; the polyethylene glycol 12 hydroxystearic acid 15 is preferably polyethylene glycol
HS 15; the above-mentioned
The weight part of the HS 15 is preferably 385-395 parts, more preferably 388-392 parts, and most preferably 390.5 parts; the weight part of the egg yolk lecithin is preferably 5-55 parts, more preferably 25-35 parts, and most preferably 29.5 parts. The source of the egg yolk lecithin is not particularly limited in the present invention, and any commercially available product that is conventional in the art may be used. In the present invention, the egg yolk lecithin functions as an emulsifier.
In the invention, the honokiol self-emulsifying microemulsion preparation comprises 290-330 parts of co-emulsifier, preferably 300-310 parts, and more preferably 306 parts; the coemulsifier is preferably absolute ethyl alcohol. The source of the absolute ethyl alcohol is not particularly limited in the invention, and a commercially available product which is conventional in the art can be used.
In the invention, the honokiol self-emulsifying microemulsion preparation comprises 4-8 parts of antioxidant, preferably 5-7 parts, and more preferably 6 parts. In the present invention, the antioxidant is preferably acetylcysteine. The source of acetylcysteine in the present invention is not particularly limited, and any commercially available product that is conventional in the art may be used.
In the present invention, the above-mentioned raw materials are all injection grade raw materials.
The invention provides a preparation method of the honokiol self-emulsifying microemulsion preparation, which comprises the following steps: 1) mixing egg yolk lecithin, an antioxidant and an auxiliary emulsifier, and mixing with polyethylene glycol 12 hydroxystearic acid 15 and an oil phase to obtain a solvent solution; 2) mixing honokiol with the solvent solution to obtain the honokiol self-emulsifying microemulsion preparation.
In the present invention, the preparation of the honokiol self-emulsifying microemulsion preparation is preferably carried out under a ten thousand grade aseptic condition. Firstly, mixing egg yolk lecithin, an antioxidant and an auxiliary emulsifier; in the present invention, the stirring is preferably accompanied during the mixing, and the rotation speed of the stirring is not particularly limited, and it is preferable to achieve sufficient dissolution. Mixing the mixed solution, polyethylene glycol 12 hydroxystearic acid 15 and an oil phase to obtain a solvent solution; in the present invention, stirring is preferably carried out during the process of obtaining the solvent solution, and the rotation speed of the stirring is not particularly limited in the present invention, and it is preferable to achieve sufficient dissolution.
After the solvent solution is obtained, mixing honokiol with the solvent solution to obtain a honokiol self-emulsifying microemulsion preparation; in the invention, the process of obtaining the honokiol self-emulsifying microemulsion preparation is preferably accompanied by stirring, and the rotating speed of the stirring is not particularly limited in the invention, so that the sufficient dissolution is preferably realized.
In the present invention, the mixing of the honokiol and the solvent solution preferably further comprises filtration with a filter membrane; the pore size of the filter membrane is preferably 0.22 μm; the filter membrane is preferably an organic filter membrane. The invention preferably also comprises the step of filling nitrogen and encapsulating after the filtration of the filter membrane; the nitrogen filling and encapsulating steps are not particularly limited, and the conventional nitrogen filling and encapsulating steps in the field can be adopted.
The invention also provides an injection preparation comprising the self-emulsifying microemulsion preparation, and the injection preparation further comprises 5% glucose injection or 0.9% NaCl injection. In the invention, the 5% glucose injection or the 0.9% NaCl injection is preferably a commercially available injection-grade product, and the content of the 5% glucose injection or the 0.9% NaCl injection in the injection preparation is not particularly limited and is determined according to the specific use concentration requirement of honokiol.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Honokiol-containing self-emulsifying microemulsion preparation18H18O2) Is 95.0-105.0% of the labeled amount.
The specification of the honokiol self-emulsifying microemulsion preparation in the embodiment is 1.0 g; the self-microemulsion injection contains honokiol (C)18H18O2)50mg。
TABLE 1 Unit dose of the formulation composition of honokiol self-microemulsion injection
The preparation method comprises the following steps:
weighing low-endotoxin lecithin and acetylcysteine according to the prescription amount under the ten-thousand-level sterile condition, adding absolute ethyl alcohol, and stirring for dissolving; adding low endotoxin into lecithin anhydrous ethanol solution
HS 15 and medium-chain triglyceride (MCT) are stirred and mixed evenly; finally adding honokiol, stirring and completely dissolving;
detecting the content of honokiol (which is 95.0-105.0% of the marked amount), and filtering with a microporous organic filter membrane with the pore diameter of 0.22 mu m under the condition of ten thousand grades of aseptic local parts after the honokiol is qualified;
and filling nitrogen and encapsulating.
Evaluation of emulsification Effect and study of compatibility of preparation
In clinical use, the self-emulsifying microemulsion preparation provided by the invention can adjust different administration concentrations according to the administration requirements of patients, so that different dilution times are designed to verify the emulsifying effect. The emulsion-forming effect, the appearance of the emulsion and the size and distribution of the particle size were observed by diluting with 5% glucose injection and 0.9% NaCl injection at oil/water ratios of 1:100 and 1: 250. As shown in tables 2 and 3, stable and completely clear nano-emulsion was rapidly formed by diluting with different amounts of 5% glucose injection or 0.9% NaCl injection, the particle size and PDI were both good, and after 8h emulsification, the particle size and PDI were still uniform and were not significantly changed.
When the diluent of the self-emulsifying micro-emulsion preparation is placed at room temperature for 1 month, no obvious abnormality is found, which indicates that the self-emulsifying micro-emulsion preparation or the diluted injection has good stability.
TABLE 25 evaluation of the emulsifying Effect of different dilutions of dextrose injection
TABLE 30.9% NaCl injection dilution different times of the evaluation of the emulsification Effect
Example 2
Pilot scale production of honokiol self-emulsifying microemulsion preparation and drug stability detection
3 pilot production tests were carried out by a pharmaceutical factory entrusted with the qualification of GMP production. The results, evaluated for the quality of three pilot production runs (runs: 20110601, 20110602, 20110603), show that: the HP15 microemulsion injection prepared by the prescription process has qualified quality, stable indexes and good repeatability.
The specific number is as follows:
scale of sample preparation: 20110601, 20110602 and 20110603 were pilot scale batches, including 739 at 20110601, 705 at 20110602 and 697 at 201100601.
And (4) investigating items: appearance (before and after emulsification), visible foreign matters before emulsification, acidity, self-emulsifying microemulsion particle size, emulsification stability for 8h, main drug content and related substances.
And (5) diluting with glucose.
First, influence factor test
(1) Test by intense light irradiation
Lofting date: 2011.8.20
Sampling and measuring date: 2011.8.20 (day 0), 2011.8.24 (day 5), 2005.8.29 (day 10)
The placing conditions are as follows: intense light irradiation (4500Lx)
Irradiating a production sample by strong light, and observing and measuring parameters such as appearance color, particle size, content and the like; the results are as follows:
TABLE 4 summary of influence factors (strong light) of Honokiol microemulsion injection
(2) High temperature test
Sample batch number: 20110601, 20110602, 20110603
Lofting date: 2011.8.30
Sampling and measuring date: 2011.8.20 (day 0), 2011.9.4 (day 5), 2011.9.9 (day 10) standing conditions: high temperature (40 ℃ C.)
TABLE 5 summary of the influence factors (high temperature) of the Honokiol microemulsion injection
(3) Accelerated test
Sample batch number: 20110601, 20110602, 20110603
Lofting date: 2011.9.25
Sampling and measuring date:
2011.9.26 (month 0), 2011.10.30 (month 1), 2011.11.26 (month 2), 2011.12.28 (month 3), 2011.3.21 (month 6)
The placing conditions are as follows:
because the influence factor test shows that the product is unstable at high temperature, the condition of 25 +/-2 ℃ is adopted to carry out accelerated experimental investigation.
TABLE 6 summary of the 25 deg.C accelerated test results for Honokiol microemulsion injections
(4) Long term test
Sample batch number: 20110601, 20110602, 20110603
Lofting date: 2011.9.25
Sampling and measuring date:
2011.9.26 (month 0), 2011.12.28 (month 3), 2012.3.22 (month 6)
The placing conditions are as follows: 4 deg.C
Packaging the three batches of samples according to the market packaging condition, placing the samples in a refrigerating chamber at 4 ℃, and measuring related indexes at 0, 3, 6, 9, 12, 18, 24 and 36 months until 6 months are detected. The results are shown in Table 7.
TABLE 7 summary of the 4 deg.C long-term sample retention test results for Honokiol microemulsion injections
The honokiol can be slightly precipitated from the microemulsion injection (before emulsification) at low temperature, and can be re-dissolved into yellow clear viscous liquid at room temperature without affecting the self-emulsifying property.
From the above tests, it can be seen that: the self-emulsifying microemulsion injection preparation prepared by the invention has no obvious change in the aspects of appearance color (before and after emulsification), pH value, related substances, stability of emulsification for 8h, main drug content and the like under the conditions of illumination and high temperature of 40 ℃, and has stable property compared with 0 day; the temperature of 40 ℃ is selected as a high-temperature condition, and the oil solution is easy to oxidize and deteriorate at high temperature, so that the high temperature is avoided in the storage process; the results of the accelerated test at 25 ℃ for 6 months show that: compared with the self-emulsifying microemulsion injection preparation in 0 month, the self-emulsifying microemulsion injection preparation prepared by the invention has no obvious changes in aspects of appearance color (before and after emulsification), pH value, related substances, emulsification stability for 8h, main drug content and the like, and has stable property; the long-term test result shows that: in a detected sample batch (6 months), the properties, the content, the related substances, the stability after emulsification and the pH value of the self-emulsifying microemulsion injection preparation prepared by the invention are not obviously changed, and the self-emulsifying microemulsion injection preparation prepared by the invention has stable property at 4 ℃.
Example 3
Safety research of self-emulsifying microemulsion preparation for intravenous injection of honokiol
According to the requirements of national new drug approval, the safety of the preparation is systematically evaluated, and the results are as follows:
1. acute toxicity test: (1) ICR mice tail vein injection and single administration toxicity test of magnolol self-emulsifying microemulsion injection: after ICR mice are injected with honokiol self-emulsifying microemulsion injection through tail veins, each animal of administration groups has toxic reaction which is mainly characterized by struggling during administration, disappearance of turnover and positive reflection after administration, weakness of limbs, prostration and the like; each administration group has animal death, and dying animal has symptoms of abdominal respiration, hind limb stiffness, convulsion, urinary incontinence, etc., 100 mg/kg-1All animals died about 15 seconds after administration, 41 mg.kg-1The group surviving animals returned to normal substantially 20 minutes or so after dosing; the weight and the food intake of the animals which survive the administration are measured, and no obvious trend change is seen; carrying out gross anatomical examination on the dead mice after administration, and finding out gray patches with different degrees of visibility on the surfaces of part of animals 'livers, gray patches with different degrees of visibility on the surfaces of part of animals' kidneys and bleeding of the lungs in different degrees; histopathological examination shows that the medicine can cause the degeneration of the liver cells of the animals to different degrees and the change of the permeability of the capillary vessels of the walls of the glomerulus and the alveolar wall to a certain degree.
At the end of the observation period, all surviving animals were examined by gross dissection, and no abnormalities of organs such as heart, liver, spleen, lung, kidney, and stomach and intestine were observed. Calculated by the regular Bliss method: the LD50 of the Honokiol self-emulsifying microemulsion injection given by ICR mouse through single tail vein injection is 50.5mg kg-1The 95% confidence interval is 45.1-56.5 mg/kg-1(ii) a LD5 is 34.7mg kg-1LD90 is 67.6 mg.kg-1。
And (4) conclusion: the LD50 of the honokiol self-emulsifying microemulsion injection given by ICR mouse single tail vein injection is 50.5mg kg-1. After administration to animals, the main toxic reactions are disappearance of righting reflex, weakness of limbs, prostration and the like.
(2) Beagle dog intravenous injection and single dose toxicity test of magnolol self-emulsifying microemulsion injection: the results show that: 100, 66.7, 44.4 and 19.8 mg/kg of honokiol self-emulsifying microemulsion injection for Beagle dog intravenous injection-1The animals in the group showed toxic reaction during the administration process, which was manifested by salivation, redness of skin in mouth and claw, wrinkle and swelling of skin in eye socket and mouth, vomiting, and paralysis of limbs, etc., at 100 mg/kg-1The animals in the group died at the time of administration, 8.8, 3.9 mg/kg-1No abnormality was observed in the group animals after administration. After administration, the ALT in the animal serum is increased in an effective relationship, which indicates that the medicine may damage the liver of the animal; 66.7, 44.4mg kg-1The serum BUN and Cr of the animals are reversibly slightly increased, which indicates that the drugs may move relativelyThe substance kidney has certain influence; TG increased in each group of animals after administration compared to before administration and the index substantially recovered 15 days after administration, which may be due to the higher concentration of lipid in the drug. After administration, WBC reversibly increased in each group of animals, and no abnormality was observed in the electrocardiographic examination. The body temperature of the animals has no obvious abnormal change after the administration. 100 mg/kg-1Group animals died, 66.7mg kg-1The animals survive, so the approximate lethal dose range of the honokiol self-emulsifying microemulsion injection given by the Beagle dog by intravenous injection under the experimental condition is 66.7-100 mg kg-1. And (4) conclusion: the approximate lethal dose range of the Beagle dog intravenous injection honokiol self-emulsifying microemulsion injection is 66.7-100 mg.kg-1. The major toxic effects in animals at high doses were salivation, redness of the skin in the mouth and claw, swelling of the skin folds in the eye socket and mouth, vomiting, flaccidity of the extremities, etc.
2. Chronic toxicity test: (1) repeated administration toxicity test of SD rat tail vein injection and magnolol self-emulsifying microemulsion injection for 30 days: the appearance, behavior and the like of animals in each administration group are not abnormal during the test period; the weight and the food intake of the animals after the administration have no significant difference compared with the control group; administration for 30 days, 2.5 mg/kg-1RBC, HGB, HCT of group female mice are reduced compared with the control group, K+The change disappears at the end of the recovery period when the level is higher than that in the control group; AST, CK and LDH of female mice of the administration group are reduced at the end of the recovery period, BUN of male mice of the administration group is reduced, but the reduction of the indexes has no obvious toxicological significance; blood coagulation index and urine index detection shows no statistical change related to administration. Administration for 30 days, 2.5 mg/kg-1The spleen weight of the group female animals is increased, the liver weight and organ coefficient of the administration group male animals are increased at the end of the convalescent period, and the adrenal weight and organ coefficient are decreased. Histopathological examination is carried out at the end of 30-day and 14-day recovery periods of administration, part of injection sites (tail veins) of animals in each administration group generate degeneration and/or necrosis of vascular endothelial cells and change of vascular wall structures, the main lesion is vascular endothelial cell swelling, and similar lesions of other organ vessels of the animals are not seen, which indicates that the medicament has a certain stimulation effect on the blood vessels of the injection sites. And (4) conclusion: under the experimental conditions, SD rats are continuously treatedThe basic nontoxic reaction dose of the self-emulsifying microemulsion injection for tail vein injection and honokiol is 500 mug.kg for 30 days-1This corresponds to 62.5 times the clinically planned dose. The medicine may have certain irritation to blood vessel of injection site, and should be paid attention during clinical medication.
(2) Beagle dog intravenous injection and repeated administration toxicity test of magnolol microemulsion injection for 30 days: the appearance, behavior and the like of animals in each administration group are not abnormal during the test period; the weight, the body temperature and the food intake of the animals after the administration have no significant difference compared with the control group; electrocardiographic examination was carried out at 15 days, 30 days and the end of the recovery period, and no abnormal change associated with drug toxicity was observed in each animal of the administration group as compared with the control group. The administration is 30 days, 1250 mug/kg-1Group animals had elevated CK compared to control group (375 + -37 × vs278 + -59, U/l, p < 0.01); administration is carried out for 15 days and 30 days, 1250 mug. kg-1The APTT of the group animals is prolonged compared with the control group (10.7 +/-1.31, vs9.7 +/-0.9, s, p is 0.141; 11.0 +/-1.3, vs9.8 +/-1.6, s, p is 0.166), but the APTT of the group animals has no statistical difference. In addition to the above changes, the individual hematology, blood biochemistry and blood coagulation indexes of the animals in each administration group have statistical differences compared with the control group, but no dose response relationship exists, and the results are caused by the individual differences of the animals and are not related to the administration. The blood electrolyte and urine tests at 15 days and 30 days after administration and at the end of the recovery period show that the animals in each administration group have no abnormal change compared with the control group. Gross anatomy examination is carried out at the end of 30 days of administration and recovery period, and no obvious abnormality is seen in animals of each administration group and control group; the weight and organ coefficient of each detected organ of each administration group animal are not obviously different from those of the control group; histopathological examination was performed at the end of 30-day and 14-day recovery periods, and lesions of liver, lung, injection site, kidney, spleen, bladder and other organs were detected, and the incidence of each administration group was approximately the same as that of the control group, and it was judged that the administration was not related.
And (4) conclusion: under the experimental condition, the Beagle dog is intravenously injected with honokiol self-emulsifying microemulsion injection for 30 days, and the non-toxic reaction dose is 250 mug.kg-1. The results suggest that Beagle dogs are 1250. mu.g.kg-1Continuous dosing may result in reversible CK elevation and slight prolongation of APTT in animals。
3. The hemolytic test of the honokiol self-emulsifying microemulsion injection comprises the following steps: the honokiol self-emulsifying microemulsion injection comprises various dosage tubes and a negative control tube (0.9% sodium chloride injection), wherein the temperature is kept at 37 ℃ for 4 hours, no hemolysis and coagulation reaction occurs, and the positive control tube (water for injection) has hemolysis within 15 minutes. And (4) conclusion: under the laboratory condition, the honokiol self-emulsifying microemulsion injection does not have hemolysis and coagulation, and meets the requirement of safety inspection of intravenous injection drugs.
4. The blood vessel irritation test of the honokiol self-emulsifying microemulsion injection comprises the following steps: after the test solution is injected into the ear vein of the rabbit, the injection parts of the control group and the test group are visually checked to have the phenomenon of macroscopic mild hyperemia and gradually recover in the observation period. The pathological tissue examination result shows that no obvious abnormality is found in the test sample group.
And (4) conclusion: under the condition of the laboratory, the honokiol self-emulsifying microemulsion injection is injected with the concentration of 5.0 mu g/ml and the administration volume of 5.0ml/kg, and does not generate irritation reaction on blood vessels.
5. And (3) carrying out a whole body active anaphylaxis test on the honokiol self-emulsifying microemulsion injection. When the honokiol self-emulsifying microemulsion injection is administrated by intravenous injection, all 6 animals in the positive control group generate extremely strong positive reactions, which shows that the whole body active anaphylaxis test is established under the experimental condition, and all the animals in the 250 mu g/kg dosage group and the 50 mu g/kg dosage group do not generate anaphylactic reactions. And (4) conclusion: under the condition of the laboratory, the honokiol self-emulsifying microemulsion injection does not cause the general active allergic reaction of the guinea pigs within the dosage range of 250 mug/kg.
6. Passive skin allergy test of honokiol self-emulsifying microemulsion injection: after being stimulated by tail vein administration, the skin is anaesthetized and taken for observation and measurement after half an hour, no blue spot is seen in the skin of 4 animals in the negative control group, and the negative control group is judged to be negative. And blue spots appear in 4 animals in the positive control group at a ratio of 1:2 in the skin, the diameters of the blue spots are larger than 0.5 mm, 2 animals at a ratio of 1: 8 in the skin have the blue spots, the diameter of 1 animal is 5.82 mm, the diameter of the other animal is 4.12 mm, and the positive rate of the positive control group is comprehensively determined to be 100%, which indicates that the test system is established. The animals in the high-dose group and the low-dose group of the test sample do not have blue spots at all points, and the animals are judged to be negative. And (4) conclusion: under the condition of the laboratory, the honokiol self-emulsifying microemulsion injection is used under the condition of the administration dosage of 250.0 mu g/kg and 50.0 mu g/kg, and passive skin anaphylactic reaction is not seen.
Study of the effect of single intravenous injection of Beagle dog and self-emulsifying microemulsion injection of magnolol on cardiovascular system and respiratory function: the Beagle dog single intravenous injection honokiol self-emulsifying microemulsion injection has no obvious difference in the indexes of animal respiration, blood pressure and electrocardio within 60 minutes after administration compared with the indexes before administration in the 25 mug.kg-1 dose group. The animals respiratory intensity, respiratory frequency, P wave time course, T wave time course, QRS time course, PR interval, QT interval and systolic pressure of the 100 mu g/kg-1 dosage group have no significant difference compared with the animals before administration within 60 minutes after administration. The diastolic blood pressure at 2 and 20 minutes of administration was statistically different from the 10 minutes prior to administration (P was 0.049, 0.045, respectively), the mean arterial blood pressure at 20 minutes of administration was statistically different from the 10 minutes prior to administration (P ═ 0.045), the heart rate at 20 minutes of administration was statistically different from the 10 minutes prior to administration (P ═ 0.046), and the diastolic blood pressure, mean arterial blood pressure, and heart rate were not significantly different from the self-control at the remaining time points. The blood pressure and heart rate of the administration group and the control group are reduced after administration, but the effects on the blood pressure and heart rate reduction are basically consistent with those of the control group. The absolute values of the diastolic pressure, mean arterial pressure and heart rate changes were small for the administration group and the control group, and the percentage of the decrease was small in magnitude.
And (4) conclusion: under the experimental condition, the single intravenous drip of Beagle dog and the self-emulsifying microemulsion injection of honokiol are at 25 mug.kg-1In time, the device has no obvious abnormal influence on the respiration, the blood pressure, the heart rate and the electrocardio of the Beagle dog. At 100. mu.g/kg-1In Beagle dogs, a slight decrease in blood pressure and heart rate may result. Therefore, it is recommended to enhance the monitoring of blood pressure and heart rate during clinical medication.
8. Study on the influence of single intravenous injection and magnolol self-emulsifying microemulsion injection on ICR mouse rod transfer test: statistical analysis shows that the concentration of the active ingredients is 100, 300 and 900 mu g/kg-1The dosage of the composition is that the incubation time of the mice 5min after the mice are transferred to the rod after the ICR mice are injected with honokiol self-emulsifying microemulsion injection for 5min by a single intravenous injection and the controlThe groups had no significant difference before and after administration (P > 0.05). And (4) conclusion: under the experimental condition, single intravenous injection of ICR mice is carried out on honokiol self-emulsifying microemulsion injection at 100, 300 and 900 mu g.kg-1When the dosage is used, the composition has no obvious influence on the movement and coordination ability of animals.
9. The effect of single intravenous injection and the honokiol self-emulsifying microemulsion injection on the autonomous activities of ICR mice is researched: through statistical analysis, the single intravenous injection of ICR mice with honokiol self-emulsifying microemulsion injection is carried out at 100, 300 and 900 mug.kg-1The number of the autonomous activities of the ICR mice was not significantly different from that of the control group before and after the administration (P > 0.05). And (4) conclusion: under the experimental condition, the single intravenous injection of the ICR mouse and the self-emulsifying microemulsion injection of honokiol are carried out at 100, 300 and 900 mu g.kg-1When the dosage is used, the number of the autonomic activities of the animals is not obviously influenced.
10. Test of Chinese hamster lung fibroblast chromosome aberration by Honokiol self-emulsifying microemulsion injection: under the two test conditions of S9 activation and non-activation, the honokiol self-emulsifying microemulsion injection is prepared in the concentration of 1.00-10.00 mg/ml-1The chromosome aberration rate is within the normal range within the concentration range. Mitomycin C (MMC, 0.05. mu.g.ml) as a direct mutagen-1) The chromosomal aberration rate is significantly increased. Indirect mutagen cyclophosphamide (CP, 5.0. mu.g.ml)-1) The chromosome aberration rate is obviously increased when S9 exists, which indicates that the test system is reliable. And (4) conclusion: under the test condition, the honokiol self-emulsifying microemulsion injection is 1.00-10.00 mg/ml-1Within the concentration range, no increase in chromosomal aberration of CHL cells was induced in the presence or absence of the S9 activation system.
ICR mouse tail vein injection and magnolol self-emulsifying microemulsion injection bone marrow eosinophilic polycythemia test: the micronucleus incidence of the positive control group is 26.6 +/-4.0 thousandths, and is obviously different from that of a blank control group (1.6 +/-0.6 thousandths) (P is less than 0.01), which indicates that the test system is reliable. Honokiol self-emulsifying microemulsion injection 25 mg/kg-1、10mg·kg-1、4mg·kg-1The micronucleus incidence rates of each dosage group at 24h time point are respectively 1.8 +/-0.8 per thousand, 1.6 +/-0.7 per thousand and 2.1 +/-0.6 per thousand; 25 mg/kg-1Dose group 48hThe micronucleus occurrence rate at the time point is 1.9 +/-0.6 per thousand which is lower than 4 per thousand, and no obvious difference (P) is generated compared with a blank control group>0.05). The ratio of PCE/(NCE + PCE) of each administration group was not lower than 20% of that of the blank control group. And (4) conclusion: under the test condition, 4-25 mg-kg of honokiol self-emulsifying microemulsion injection is injected into tail vein of ICR mouse-1And has no function of causing marrow hyperpigmentation red cell micronucleus formation.
12. Histidine-deficient salmonella typhimurium back mutation test by using honokiol self-emulsifying microemulsion injection:
in the honokiol self-emulsifying microemulsion injection, the reversion colony numbers generated by TA97, TA98, TA100, TA102 and TA1535 are induced to have no significant difference (P is more than 0.05) compared with a blank control under the two test conditions of S9 activation and non-activation within the concentration range of 0.5-5000 mug/dish; under the activation condition of S9, after the sample and the mixed solution of S9 are added at the concentration of 5000 mug/dish, the flat plate is milk white, and accurate counting can not be carried out when the result is counted, but almost no increase of the strain return variable is observed. The result shows that the honokiol self-emulsifying microemulsion injection has no gene mutation effect on five test strains within the test concentration range of 0.5-5000 mug/dish. Direct mutagens 4-NQO (2.0. mu.g/dish), MMC (0.5. mu.g/dish), NaN3(1.5 mu g/dish) and the indirect mutagen 2-AF (60 mu g/dish), cis-platinum (50 mu g/dish) and CP (200 mg/dish) under the corresponding test conditions, the numbers of the reversion colonies generated by the five strains are obviously more than that of a blank control (P < 0.01), and the used test system is reliable. And (4) conclusion: under the test condition, the honokiol self-emulsifying microemulsion injection has no mutagenicity to salmonella typhimurium strains TA97, TA98, TA100, TA102 and TA1535 in the concentration range of 0.5-5000 mug/dish, no matter whether an S9 activation system exists or not.
Example 4
Honokiol-containing self-emulsifying microemulsion preparation18H18O2) Is 95.0-105.0% of the labeled amount. The specification of the honokiol self-emulsifying microemulsion preparation in the embodiment is 1.0 g; the self-microemulsion injection contains honokiol (C)18H18O2)50 mg; the compositions are shown in Table 8.
Table 8 the formulation composition of honokiol self-microemulsion injection per unit dose in this example
The preparation process comprises the following steps:
adding honokiol into 1, 2-propylene glycol, stirring for dissolving, adding dibutyl phenol, stirring for dissolving; then mixing the mixed solution of polyethylene glycol 12, hydroxystearic acid 15 and ethyl oleate, stirring and mixing until the mixture is clear, thus obtaining the honokiol self-emulsifying solution. Filtering the above solution with 0.22 μm microporous membrane, aseptically filling nitrogen, and packaging to obtain self-emulsifying microemulsion preparation for injection.
Example 5
Honokiol self-emulsifying microemulsion preparation containing Honokiol (C)18H18O2) Is 95.0-105.0% of the labeled amount. The specification of the honokiol self-emulsifying microemulsion preparation in the embodiment is 1.0 g; the self-microemulsion injection contains honokiol (C)18H18O2)50 mg; the compositions are shown in Table 9.
Table 9 composition of the formula of the honokiol self-microemulsion injection per unit dose in this example
The preparation process comprises the following steps:
adding honokiol into anhydrous ethanol, stirring for dissolving, adding tert-butyl p-hydroxyanisole and ascorbyl palmitate, stirring for dissolving; then mixing the mixed solution of 15 parts of polyethylene glycol 12 hydroxystearic acid, 80 parts of tween and isopropyl palmitate, stirring and mixing until the mixture is clear, and obtaining the honokiol self-emulsifying solution.
Filtering the above solution with 0.22 μm microporous membrane, aseptically filling nitrogen, and packaging to obtain self-emulsifying microemulsion preparation for injection.
Example 6
Honokiol self-emulsifying microemulsion preparation containing honokiolHackberry phenol (C)18H18O2) 95.0 to 105.0% of the labeled amount. The specification of the honokiol self-emulsifying microemulsion preparation in the embodiment is 1.0 g; the self-microemulsion injection contains honokiol (C)18H18O2)50 mg; the compositions are shown in Table 10.
Table 10 composition of the formula of the honokiol self-microemulsion injection per unit dose in this example
The preparation process comprises the following steps:
adding honokiol into anhydrous ethanol, stirring for dissolving, adding tert-butyl p-hydroxyanisole, stirring for dissolving; adding vitamin C into polyethylene glycol 200, stirring and dissolving; mixing the two solutions uniformly; then adding the mixed solution of pluronic F68, polyoxyethylene ether 40 hydrogenated castor oil, poloxamer 188 and isopropyl myristate, stirring and mixing until the mixture is clear, thus obtaining the honokiol self-emulsifying solution.
Filtering the above solution with 0.22 μm microporous membrane, aseptically filling nitrogen, and packaging to obtain self-emulsifying microemulsion preparation for injection.
Example 7
Honokiol self-emulsifying microemulsion preparation containing Honokiol (C)18H18O2) Is 95.0-105.0% of the labeled amount. The specification of the honokiol self-emulsifying microemulsion preparation in the embodiment is 1.0 g; the self-microemulsion injection contains honokiol (C)18H18O2)50 mg; the compositions are shown in Table 11.
TABLE 11 formulation composition of honokiol self-microemulsion injection per unit dose in this example
The preparation process comprises the following steps:
adding honokiol into anhydrous ethanol and polyethylene glycol 400, stirring for dissolving, adding tert-butyl p-hydroxyanisole and propyl gallate, stirring for dissolving; then adding a mixed solution of polyethylene glycol 12 hydroxystearic acid 15, polyoxyethylene ether 35 hydrogenated castor oil and isopropyl myristate, stirring and mixing until the solution is clear, and obtaining the honokiol self-emulsifying solution.
Filtering the above solution with 0.22 μm microporous membrane, aseptically filling nitrogen, and packaging to obtain self-emulsifying microemulsion preparation for injection.
Example 8
Honokiol self-emulsifying microemulsion preparation containing Honokiol (C)18H18O2) 95.0 to 105.0% of the labeled amount. The specification of the honokiol self-emulsifying microemulsion preparation in the embodiment is 1.0 g; the self-microemulsion injection contains honokiol (C)18H18O2)50 mg; the compositions are shown in Table 12.
Table 12 composition of the formula of honokiol self-microemulsion injection per unit dose in this example
Preparation process
Adding honokiol into anhydrous ethanol and glycerol, stirring for dissolving, adding acetylcysteine and alpha-tocopherol, and stirring for dissolving; then adding the mixed solution of polyethylene glycol 12 hydroxystearic acid 15, soybean lecithin and medium chain triglyceride, stirring and mixing until the mixture is clear, and obtaining the honokiol self-emulsifying solution.
Filtering the above solution with 0.22 μm microporous membrane, aseptically filling nitrogen, and packaging to obtain self-emulsifying microemulsion preparation for injection.
Comparative example 1
Prepared from microemulsion injection according to the composition shown in Table 13
TABLE 13 formulation composition of honokiol self-microemulsion injection per unit dose in this example
Preparation process
Adding honokiol into anhydrous ethanol, stirring for dissolving, adding acetylcysteine, and stirring for dissolving; then, a mixture of polyethylene glycol 12 hydroxystearic acid 15 and medium-chain triglyceride was added thereto, and the mixture was stirred and mixed, but a uniform clear solution could not be obtained, and turbidity and precipitation occurred.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (4)
1. A honokiol self-emulsifying microemulsion preparation for injection is characterized by comprising the following raw materials: 50mg of honokiol, 218mg of oil phase, 420mg of emulsifier, 306mg of coemulsifier and 6mg of antioxidant;
the oil phase is a medium chain triglyceride;
the emulsifier is
HS 15 and egg yolk lecithin; the described
390.5mg of HS 15, 29.5mg of egg yolk lecithin;
the auxiliary emulsifier is absolute ethyl alcohol;
the antioxidant is acetylcysteine.
2. The preparation method of the honokiol self-emulsifying microemulsion preparation for injection as claimed in claim 1, comprises the following steps:
1) mixing egg yolk lecithin, antioxidant and auxiliary emulsifier, and mixing with the mixture
Mixing HS 15 and an oil phase to obtain a solvent solution;
2) mixing honokiol with the solvent solution to obtain the honokiol self-emulsifying microemulsion preparation.
3. The method of claim 2, wherein the mixing of the honokiol and the solvent solution in step 2) further comprises filtration with a filter membrane; the aperture of the filter membrane is 0.22 um.
4. An injection preparation comprising the self-emulsifying microemulsion preparation for injection as set forth in claim 1, which further comprises a 5% glucose injection or a 0.9% NaCl injection.
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Citations (3)
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| CN102302451A (en) * | 2011-09-13 | 2012-01-04 | 郑州大学 | Honokiol submicroemulsion injection and production method thereof |
| CN103083237A (en) * | 2013-01-21 | 2013-05-08 | 郑州大学 | Lipid emulsion of magnolol and preparation method thereof |
| CN105726521A (en) * | 2014-12-10 | 2016-07-06 | 瑞普（天津）生物药业有限公司 | Emulsion containing magnolol or honokiol, and preparation method of emulsion |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102302451A (en) * | 2011-09-13 | 2012-01-04 | 郑州大学 | Honokiol submicroemulsion injection and production method thereof |
| CN103083237A (en) * | 2013-01-21 | 2013-05-08 | 郑州大学 | Lipid emulsion of magnolol and preparation method thereof |
| CN105726521A (en) * | 2014-12-10 | 2016-07-06 | 瑞普（天津）生物药业有限公司 | Emulsion containing magnolol or honokiol, and preparation method of emulsion |
Non-Patent Citations (1)
| Title |
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| 和厚朴酚自微乳化给药系统的研究;甘露等;《第三军医大学学报》;20131015;第35卷(第19期);第2073-2076页 * |
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