CN108553417B

CN108553417B - Osthole self-emulsifying drug release system and preparation method and application thereof

Info

Publication number: CN108553417B
Application number: CN201810298063.8A
Authority: CN
Inventors: 张小飞; 果秋婷; 史亚军; 邹俊波; 王媚; 孙静; 郭东艳; 王晶; 王昌利; 唐志书; 崔春利; 程江雪
Original assignee: Shaanxi University of Chinese Medicine
Current assignee: Shaanxi University of Chinese Medicine
Priority date: 2018-03-30
Filing date: 2018-03-30
Publication date: 2022-06-21
Anticipated expiration: 2038-03-30
Also published as: CN108553417A

Abstract

The invention provides an osthole self-emulsifying drug release system, which is a preparation prepared from osthole and auxiliary materials, wherein the osthole accounts for 0.1-10% by mass, and the auxiliary materials account for 90-99.9% by mass; the auxiliary material comprises an oil phase, a surfactant and a cosurfactant, wherein the oil phase, the surfactant and the cosurfactant account for the auxiliary material in percentage by mass respectively as follows: 25-55% of oil phase, 5-45% of surfactant and the balance cosurfactant. The invention also provides a preparation method and application of the osthole self-emulsifying drug delivery system. The osthole self-emulsifying drug delivery system provided by the invention can be quickly emulsified after contacting with water, the obtained emulsion droplets have small average particle size (31.8 +/-1.6 nm), can be quickly dissolved out in vitro and have high dissolution rate, and a patient only needs to add proper water for dilution before use and spray (mist) on the surface of the skin, so that the operation is simple, and the compliance of the patient is greatly improved. The invention improves the solubility of the osthole by the preparation technology, improves the bioavailability of the osthole in human body and has better clinical application prospect.

Description

Osthole self-emulsifying drug release system and preparation method and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to an osthole self-emulsifying drug delivery system, and a preparation method and application thereof.

Background

Osthole (osthol) belongs to alkyl coumarin compounds, is an effective component extracted and separated from dried fruit fructus Cnidii (cnidiummonieril. cuss) of cnidium (osthol) of Umbelliferae, and is mainly used for treating skin diseases such as eczema, tinea of feet and hands, urticaria, etc. clinically. In recent years, studies show that osthole has clinical effects of improving arrhythmia, resisting hypertension, inhibiting tumor cell growth, relieving pain and inflammation, inhibiting allergy and the like.

Due to poor water solubility of osthole, the drug absorption is poor, and the clinical treatment effect cannot be achieved. Therefore, it is necessary to improve the solubility of osthole by the preparation technology, increase the absorption rate and degree of the medicine and achieve good clinical treatment effect.

Disclosure of Invention

In order to solve the problems, the invention provides an osthole self-emulsifying drug delivery system.

The osthole self-emulsifying drug delivery system is a preparation prepared from osthole and auxiliary materials, wherein the osthole accounts for 0.1-10% by mass, and the auxiliary materials account for 90-99.9% by mass; the auxiliary material comprises an oil phase, a surfactant and a cosurfactant, wherein the oil phase, the surfactant and the cosurfactant account for the auxiliary material in percentage by mass respectively as follows: 25-55% of oil phase, 5-45% of surfactant and the balance cosurfactant.

Wherein the mass ratio of the osthole to the auxiliary materials is as follows: osthole 1% and adjuvant 99%.

Wherein in the auxiliary materials, the oil phase, the surfactant and the cosurfactant account for the following components in percentage by mass: 35-55% of oil phase, 15-35% of surfactant and 15-30% of cosurfactant.

Further, in the auxiliary materials, the oil phase, the surfactant and the cosurfactant respectively account for the following components in percentage by mass: 40-50% of oil phase, 25-35% of surfactant and 20-25% of cosurfactant.

Further, in the auxiliary materials, the oil phase, the surfactant and the cosurfactant respectively account for the following components in percentage by mass: 46.9% of oil phase, 29.9% of surfactant and 23.2% of cosurfactant.

Wherein the oil phase is one or a combination of propylene glycol monocaprylate, glyceryl monocaprylate and ethyl oleate.

Further, the oil phase is glyceryl monocaprylate.

The surfactant is one or a combination of more of polyoxyethylene castor oil, polyethylene glycol 40 stearate, caprylic capric polyethylene glycol glyceride, Tween20 and Tween 80.

Further, the surfactant is Tween 20.

Wherein, the cosurfactant is one or a combination of two of propylene glycol and PEG 400.

Further, the co-surfactant is PEG 400.

Wherein, the preparation is an external preparation, preferably, the preparation is a liquid preparation and a soft capsule.

The invention also provides an osthole emulsion, which is characterized in that: it is a preparation formed by mixing the osthole self-emulsifying drug delivery system and water.

Furthermore, the osthole emulsion is a preparation formed by diluting the osthole self-emulsifying drug release system by 100 times with water.

The invention also provides a preparation method of the osthole self-emulsifying drug delivery system, which comprises the following steps:

mixing the oil phase, the surfactant and the cosurfactant according to the prescription amount, and adding the osthole to dissolve under stirring to obtain the osthole.

The invention also provides a preparation method of the osthole emulsion, which comprises the following steps:

diluting osthole self-emulsifying drug delivery system with water.

The invention also provides the application of the osthole self-emulsifying drug delivery system in preparing drugs for treating skin diseases such as eczema, tinea of feet and hands, urticaria and the like.

The invention also provides the application of the osthole emulsion in preparing medicaments for treating skin diseases such as eczema, tinea of feet and hands, urticaria and the like.

Self-emulsifying drug delivery system (SEDDS) is an isotropic mixture of an oil phase, a surfactant and a co-surfactant and containing a drug, which spontaneously forms an isotropic, transparent or translucent thermodynamically stable liquid dispersion (e.g., microemulsion) with an average particle size of between 20 and 500nm upon contact with an aqueous medium and mild agitation. The self-emulsifying medicine releasing system is one self-emulsifying medicine feeding system with emulsified liquid drop size smaller than 100 nm.

The osthole self-emulsifying drug delivery system provided by the invention is quickly microemulsified after contacting with water, the average particle size of the obtained emulsion drop is small (31.8 +/-1.6 nm), and the in vitro dissolution is quick and high. The osthole self-emulsifying drug delivery system provided by the invention has the advantages that a patient only needs to add proper water for dilution before use and spray (mist) on the surface of the skin, the operation is simple, and the compliance of the patient is greatly improved. The invention improves the solubility of the osthole by the preparation technology, improves the absorption speed and the degree of the osthole, namely improves the bioavailability of the osthole in human body, and has better clinical application prospect.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

FIG. 1 is a particle size distribution and Zeta potential diagram of osthole microemulsion after the osthole self-emulsifying drug delivery system of the invention self-emulsifies

FIG. 2 is a transmission electron microscope photograph of the osthole microemulsion after the osthole self-emulsifying drug delivery system of the invention self-emulsifies

FIG. 3 shows the osthole self-emulsifying drug delivery system and the in vitro dissolution curve of osthole bulk drug (n ═ 6)

Detailed Description

EXAMPLE 1 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

Taking the formula amount of glyceryl monocaprylate, Tween20 and PEG400, mixing uniformly, adding osthole under stirring, continuing stirring for 30min to fully dissolve osthole, and packaging to obtain the final product.

EXAMPLE 2 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

EXAMPLE 3 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

EXAMPLE 4 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

EXAMPLE 5 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

Example 6 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

Taking the formula amount of glyceryl monocaprylate, Tween20 and PEG400, mixing uniformly, adding osthole under stirring, continuing stirring for 30min to fully dissolve the osthole, and packaging to obtain the final product.

EXAMPLE 7 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

Example 8 preparation of the osthole self-emulsifying drug delivery system of the present invention

1 prescription composition

2 preparation method

Example 9 preparation of an osthole emulsion of the present invention

1 prescription composition

2 preparation method

1) Preparing a self-emulsifying drug release system: taking the glyceryl monocaprylate, Tween20 and PEG400 according to the prescription amount, uniformly mixing, adding the osthole under stirring, and continuously stirring for 30min to fully dissolve the osthole to obtain the composition;

2) preparing an emulsion: adding water to dilute the self-emulsifying drug release system obtained in the step 1) to 100 times to obtain the self-emulsifying drug release system.

The beneficial effects of the invention are illustrated by way of experimental examples below:

the self-emulsifying drug delivery system meeting the requirement of medication needs to have the following conditions: the drug has larger dissolving capacity in the self-emulsifying drug release system so as to reduce the use amount of auxiliary materials of the self-emulsifying drug release system, in particular to the use amount of a surfactant and a cosurfactant; the oil phase, the surfactant and the cosurfactant have good intermiscibility and can form a uniform, transparent and isotropic solution; the self-emulsifying drug delivery system can be quickly emulsified to form a homogeneous system after being gently stirred by adding water.

Apparatus and materials

DF-101S type heat collection type constant temperature magnetic stirrer (Steud City Zhaohuo instruments, Inc.); agilent-1100 high performance liquid chromatograph (Agilent technologies, Inc.); FA1004N analytical balance (shanghai precision instruments ltd); malvern Zetasizer (Nano ZS90) dynamic light scattering instrument (Malvern, uk); JEM-1400120 kV transmission electron microscope (Japanese Electron corporation); an RCZ-6B intelligent dissolution instrument (Shanghai yellow sea medical testing instrument factory); dialysis bag (molecular weight cut-off: 12,000 daltons, Shanghai-derived leaf Biotech Co., Ltd.).

Osthole reference (purchased from China pharmaceutical biologicals institute); osthole bulk drug (Baoji City, san Xian biological development Co., Ltd., content: 80%); propylene glycol monocaprylate (Capryol 90, trade of jiafa lion, ltd); glyceryl monocaprylate (Capmul MCM C8, trade of Jiafa lion, Inc.); ethyl oleate (basf); polyoxyethylene castor oil (Cremophor EL, basf corporation); polyethylene glycol 40 stearate (Cremophor RH 40, Pasteur applications chemical Co., Ltd.); caprylic capric polyethylene glycol glyceride (Labrasol, trade of jiafa lion, ltd.); tween 80/Tween 20 (Nanjing Will chemical Co., Ltd.); polyethylene glycol 400(PEG400, Hubei Gekko pharmaceutical adjuvant, Limited liability company), 1, 2-propylene glycol (Dache chemical reagent factory, Tianjin).

Experimental example 1 oil phase, surfactant and co-surfactant were screened by solubility experiment

And (3) determining the solubility of the osthole in each auxiliary material by adopting a high performance liquid chromatography, and determining the optimal mode of each auxiliary material.

1 chromatographic conditions

A chromatographic column: phenomenex C18 column (250 mm. times.4.6 mm,5 μm); mobile phase: methanol-water (70:30, v/v), flow rate: 1.0 mL/min^-1(ii) a Detection wavelength: 322 nm; column temperature: 25 ℃; sample introduction amount: 20 μ L.

2 preparation of control solutions

Precisely weighing 4.8mg of osthole reference substance, placing in a10 m L volumetric flask, adding methanol to dissolve and dilute to scale, and shaking up; thus obtaining 0.48mg/m L osthole reference substance stock solution; sequentially diluting the osthole reference substance stock solution with methanol to obtain osthole reference substance solutions of 0.348 mg/m L, 0.288 mg/m L, 0.192 mg/m L.

3 preparation of sample solution

The solubility of osthole in each oil phase, surfactant and co-surfactant was determined by the shake flask method. Adding 0.02g of osthole into each screw cap glass vial containing 2mL of auxiliary materials, placing the mixture on a vortex mixer for vortex mixing for 5min to fully mix the medicine and the auxiliary materials, placing the mixture in a constant-temperature shaking water bath for shaking and dissolving for 72 h to fully dissolve the medicine and reach a saturated state, then centrifuging for 10min at 5000rpm, taking supernatant, filtering with a 0.22 mu m filter membrane, taking 1mL of filtrate, and diluting with 50 times of methanol to obtain a sample solution.

4 dissolution determination

Respectively injecting the reference substance solution and the sample solution into a high performance liquid chromatograph, measuring the osthole content in each sample solution by an external standard method, and calculating the solubility of the osthole in each auxiliary material.

5 results of the experiment

The results are shown in Table 1.

TABLE 1 solubility of osthole in adjuvants (n ═ 3)

From the results of the solubility experiments, it is found that osthole has the highest solubility in oil phase Capmul MCMC8, the highest solubility in surfactant Tween20, the highest solubility in 194.34 + -0.69, the highest solubility in co-surfactant PEG400, and the highest solubility in 36.78 + -0.19.

Therefore, the preferable application of the invention is that Capmul MCMC8 is used as the oil phase of the osthole self-emulsifying drug delivery system, Tween20 is used as a surfactant, and PEG400 is used as a cosurfactant.

Experimental example 2 comparison of formula ratios of different auxiliary materials of osthole self-emulsifying drug delivery system

The percentage contents of Capmul MCMC8, Tween20 and PEG400 in the auxiliary materials are respectively used as prescription variables, the average particle size, the self-emulsifying time and the 15min drug release amount are selected as evaluation indexes, and the emulsifying effect and the drug release capacity of the osthole self-emulsifying drug release system are compared when the auxiliary materials are matched according to different prescription proportions.

Preparing auxiliary materials according to the mixture ratio shown in table 2, and mixing the osthole and the prepared auxiliary materials according to the proportion of 1%: 99 percent of the mass ratio is prepared into 11 groups of osthole self-emulsifying drug release systems. The sum of the proportions of the three components is 100 percent.

Table 2 ingredients of each adjuvant prescription and their proportions

The average particle size, the self-emulsifying time and the 15min drug release amount of the 11 prepared osthole self-emulsifying drug delivery systems are measured according to the following methods:

diluting each group of osthole self-emulsifying drug delivery system with distilled water to 100 times, and measuring the average particle diameter of formed microemulsion by using Malvern Zetasizer (Nano ZS 90);

adding 1mL of osthole self-emulsifying drug delivery system into 900mL of distilled water at 37 +/-0.5 ℃ under continuous stirring (50rpm) by using a dissolution rate determination device paddle method, and determining the time required by each group of completely uniform dispersion system, namely the self-emulsifying time of each group;

the second dissolution test method of 0931 in the four ministries of the general rules of the 2015 edition of Chinese pharmacopoeia is adopted to test the 15min drug release amount of the osthole self-emulsifying drug release system of each group (Y3). The dissolution medium was a pH 6.8 phosphate buffer containing 0.5% (w/v) Tween 80, the volume of the medium was 900mL, the temperature of the medium was (37. + -. 0.5 ℃ C.), and the stirring paddle speed was 50 rpm. Starting a dissolution instrument, adding a self-emulsifying drug delivery system which is equivalent to 50mg of osthole into a dissolution medium, sampling at 15min, centrifuging the sample at 20000 rpm for 10min, taking supernatant, and determining the drug content in the sample by using an HPLC analysis method to obtain the 15min drug release amount of the osthole self-emulsifying drug delivery system.

The results are shown in Table 3:

TABLE 3 Effect of different adjuvant formulations on average particle size, self-emulsification time and 15min drug release

The above experimental results show that:

1) when the mass ratio of Capmul MCMC8 (oil phase) in the auxiliary material prescription is more than 55% (prescriptions 10, 11, 12 and 13), the 15min drug release amount of the obtained self-emulsifying drug release system is less than 95%, and is reduced along with the increase of the mass ratio of Capmul MCMC8 (oil phase) in the auxiliary material prescription, in particular, when the mass ratio of Capmul MCMC8 (oil phase) in the auxiliary material prescription reaches 80% (prescription 12), the 15min drug release amount of the obtained self-emulsifying drug release system is only 75.2%, and when the mass ratio of Capmul MCMC8 (oil phase) in the auxiliary material prescription reaches 85.2% (prescription 13), the 15min drug release amount of the obtained self-emulsifying drug release system is only 62.0%; when the weight ratio of Capmul MCMC8 (oil phase) in the auxiliary material prescription is less than 25% (prescription 9), the self-emulsifying drug delivery system can not be emulsified to form particles when meeting water. Therefore, the weight ratio of Capmul MCMC8 (oil phase) in the auxiliary material prescription is preferably 25-55%;

2) the prescription 5 is the most preferable auxiliary material prescription, the self-emulsifying drug release system has the shortest self-emulsifying time which is only 13s, and the drug release quantity is the highest at 15min and is as high as 99.9 percent;

3) in the prescription 14, the mass ratio of the Tween20 (surfactant) in the auxiliary material prescription is 52.6% and is more than 45%, and the 15-min drug release amount of the self-emulsifying drug release system is only 72.1%, so the mass ratio of the Tween20 (surfactant) in the auxiliary material prescription is preferably 5-45%.

In conclusion, in the osthole self-emulsifying drug delivery system, the formula and the ratio of auxiliary materials are as follows: 25-55% of oil phase, 5-45% of surfactant and the balance cosurfactant. The most preferable prescription ratio of the auxiliary materials is as follows: 46.9% of oil phase, 29.9% of surfactant and 23.2% of cosurfactant.

The self-emulsifying time, the formed microemulsion appearance, the average particle size, PdI, Zeta potential, the microscopic morphology and the in vitro release condition of the optimal osthole self-emulsifying drug release system prescription of the invention are evaluated by experiments.

Experimental example 3 quality determination of osthole self-emulsifying drug delivery system

The osthole self-emulsifying drug delivery systems involved in the following experiments are all the osthole self-emulsifying drug delivery systems with the optimal formula of the invention prepared in example 1 of the invention.

1 self-emulsification time

Using a dissolution rate determination device paddle method, 1mL of the osthole self-emulsifying delivery system was added to 900mL of distilled water at 37. + -. 0.5 ℃ with continuous stirring (50rpm), the time required for complete uniform dispersion was determined, and the self-emulsifying time and appearance were recorded.

The experimental result shows that the self-emulsifying time of the osthole self-emulsifying drug delivery system is (15.2 +/-1.4) seconds, which shows that the osthole self-emulsifying drug delivery system can quickly form light blue transparent opalescent solution when contacting with water.

2 measurement of average particle diameter and Zeta potential

The osthole self-emulsifying drug delivery system is diluted 100 times with distilled water, and the mean particle size and Zeta potential of the formed microemulsion are determined using a Malvern Zetasizer (Nano ZS 90).

The measurement results are shown in FIG. 1.

The experimental result shows that the average grain diameter of the osthole microemulsion is (31.8 +/-1.6) nm, the PdI value is 0.164 +/-0.014, the lower PdI value indicates that the average grain diameter distribution is more uniform, and the Zeta potential value is-

(26.1. + -. 1.4) mV, the negative charge may be due to the presence of surfactants and co-surfactants in the formulation.

3 observation by scanning electron microscope

Diluting the osthole self-emulsifying drug release system with distilled water, dripping the microemulsion on a double-sided adhesive tape of an aluminum bar, placing a sample in a scanning electron microscope chamber, scanning the osthole microemulsion under the conditions of the resolution of 1.0nm and the accelerating voltage of 15kv, and taking an electron microscope picture.

The results of the experiment are shown in FIG. 2.

The transmission electron micrograph shows that the osthole microemulsion is round and regular spherical, and the particle diameter of most particles is about 30 nm.

4 in vitro dissolution Studies

The dissolution rate of the osthole raw material medicine and the in-vitro medicine of the osthole self-emulsifying medicine-releasing system is measured by adopting a dissolution rate measuring method of 0931 which is the general rule of the four parts of the 2015 edition of Chinese pharmacopoeia. The dissolution medium was a pH 6.8 phosphate buffer containing 0.5% (w/v) Tween 80, the volume of the medium was 900mL, the temperature of the medium was (37. + -. 0.5 ℃ C.), and the stirring paddle speed was 50 rpm. Starting a dissolution instrument, respectively taking 50mg of osthole bulk drug and a self-emulsifying drug release system equivalent to 50mg of osthole, adding the materials into a dissolution medium, respectively sampling at preset times of 0, 5, 10, 15, 30, 45, 60, 75, 90, 120, 150 and 180min, centrifuging the sample at 20000 rpm for 10min, taking supernatant, and determining the drug content in the sample by using an HPLC analysis method.

The measurement results are shown in FIG. 3.

The dissolution curve research shows that the dissolution rate of the osthole raw material drug is less than 40% within 180min, and the osthole self-microemulsifying drug release system can reach more than 90% within 45min, which shows that the self-emulsifying drug release system obviously improves the dissolution rate of the osthole.

In conclusion, the osthole self-emulsifying drug delivery system provided by the invention is rapidly micro-emulsified after contacting with water, the average particle size of the obtained emulsion drop is small (31.8 +/-1.6 nm), and the in vitro dissolution is rapid and high. The invention improves the solubility of the osthole by the preparation technology, improves the absorption speed and the degree of the osthole, namely improves the bioavailability of the osthole in human body, and has better clinical application prospect.

Claims

1. An osthole emulsion characterized in that: it is a preparation formed by diluting an osthole self-emulsifying drug delivery system by 100 times with water;

the osthole self-emulsifying drug delivery system is a preparation prepared from osthole and auxiliary materials, wherein the osthole accounts for 1% by mass, and the auxiliary materials account for 99% by mass; the auxiliary material comprises an oil phase, a surfactant and a cosurfactant, wherein the oil phase, the surfactant and the cosurfactant account for the auxiliary material in percentage by mass respectively as follows: 46.9% of oil phase, 29.9% of surfactant and 23.2% of cosurfactant; the oil phase is glyceryl monocaprylate; the surfactant is Tween 20; the cosurfactant is PEG 400;

the preparation method of the osthole self-emulsifying drug delivery system comprises the following steps: mixing the oil phase, the surfactant and the cosurfactant according to the prescription amount, and adding the osthole to dissolve under stirring to obtain the osthole.

2. The method for producing an osthole emulsion according to claim 1, characterized in that: it comprises the following steps:

diluting osthole self-emulsifying drug delivery system with water.

3. Use of the osthole emulsion of claim 1 for the preparation of a medicament for the treatment of eczema, tinea manus, tinea pedis, urticaria.