技术领域technical field
本发明涉及一种布洛芬微乳给药系统及其制备方法。The invention relates to an ibuprofen microemulsion drug delivery system and a preparation method thereof.
背景技术Background technique
通过高通量筛选获得的具有潜在开发价值的候选药物，超过40%的单体难溶于水。这些单体通常由于难溶性相关的缺陷，比如口服生物利用度低或者剂量无法逾越等，使其药物研究过早夭折（Lipinski, C.A., Lombardo, F., Dominy, B.W., Feeney, P.J., 2001. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development setting. Adv. Drug Deliv. Rev. 46(1-3), 3-26）。在药剂学领域，很多方法可以提高难溶性药物的溶解度，例如成盐、改变药物的晶型，采用混合溶剂、助溶剂、增溶剂，形成固体分散体，包合物等。或者采用新型给药系统如胶束、脂质体、微乳等。其中，微乳凭借其热力学稳定性，对难溶性药物有很好的溶解性，以及工艺简单易产业化等优点倍受研究者的青睐。Candidate drugs with potential development value obtained through high-throughput screening, more than 40% of the monomers are poorly soluble in water. These monomers are often prematurely aborted in pharmaceutical research due to poor solubility-related deficiencies, such as low oral bioavailability or insurmountable doses (Lipinski,C.A., Lombardo, F., Dominy, B.W., Feeney, P.J., 2001.Experimental and computational approaches to estimate solubility andpermeability in drug discovery and development setting. Adv. Drug Deliv. Rev. 46(1-3), 3-26). In the field of pharmacy, many methods can improve the solubility of poorly soluble drugs, such as salt formation, changing the crystal form of drugs, using mixed solvents, co-solvents, solubilizers, forming solid dispersions, clathrates, etc. Or use new drug delivery systems such as micelles, liposomes, microemulsions, etc. Among them, microemulsions are favored by researchers due to their thermodynamic stability, good solubility for insoluble drugs, and simple process and easy industrialization.
微乳 (microemulsion, ME) 是由油相、水相、表面活性剂和助表面活性剂按一定比例组成的各向同性的热力学稳定体系。由于内部同时存在亲油、亲水区域，微乳能显著增加难溶性药物的溶解度，在医药领域被广泛用于提高药物溶解度。然而，微乳形成及其毒性之间存在一个悖论，即微乳的形成通常需要大量的表面活性剂，表面活性剂的大剂量使用将导致毒性的产生，从而极大地制约了微乳的应用。表面活性剂的溶血性、组织病理学反应等毒性构成了微乳应用的一道难以逾越的技术堡垒。因此，目前只有少数微乳（或自微乳化）产品被食品药品监督管理局（Food and Drug Administration, FDA）批准。例如，专利CN103690483A公开了白花前胡甲素微乳及其制备方法。在该专利中，微乳显著提高了白花前胡甲素的溶解度（0.8%），但是，该处方中表面活性剂的总量高达30% - 50%，直接影响了白花前胡甲素微乳的临床应用。Microemulsion (ME) is an isotropic thermodynamically stable system composed of oil phase, water phase, surfactant and co-surfactant in a certain proportion. Due to the presence of both lipophilic and hydrophilic regions inside, microemulsions can significantly increase the solubility of insoluble drugs, and are widely used in the field of medicine to improve drug solubility. However, there is a paradox between the formation of microemulsions and their toxicity, that is, the formation of microemulsions usually requires a large amount of surfactants, and the use of large doses of surfactants will lead to toxicity, which greatly restricts the application of microemulsions. . The toxicity of surfactants such as hemolysis and histopathological reactions constitute an insurmountable technical fortress in the application of microemulsions. Therefore, only a few microemulsion (or self-microemulsion) products are currently approved by the Food and Drug Administration (Food and Drug Administration).and Drug Administration, FDA) approval. For example, the patent CN103690483A discloses proscartin microemulsion and its preparation method. In this patent, the microemulsion significantly increases the solubility of prosaltin (0.8%), however, the total amount of surfactant in this formulation is as high as 30% -50%, which directly affected the clinical application of proscitin microemulsion.
布洛芬属于生物药剂学分类系统II类（BCS class II）药物，属于低溶解性、高渗透性药物，溶解是其吸收的限制因素。目前报导的布洛芬微乳，其表面活性剂含量普通较高。以专利文献CN101904815 A为例，该发明使用中碳链三甘酯作为微乳的油相制备微乳可以显著增加布洛芬的溶解度 (20 mg/ml)，该微乳处方中表面活性剂用量高达30 % - 40 %，表面活性剂用量是药物的15~20倍。Ibuprofen belongs to the class II (BCS class II) drug of the biopharmaceutics classification system, which is a drug with low solubility and high permeability, and dissolution is the limiting factor for its absorption. The ibuprofen microemulsion of report at present, its surfactant content is generally higher. Take the patent document CN101904815A is example, this invention uses medium carbon chain triglyceride as the oil phase of microemulsion to prepare microemulsion and can significantly increase the solubility of ibuprofen (20mg/ml), the amount of surfactant used in this microemulsion formulation is as high as 30% - 40%, and the amount of surfactant used is 15 to 20 times that of the drug.
发明内容Contents of the invention
本发明的目的在于提供一种布洛芬微乳给药系统，其中表面活性剂的绝对用量被显著降低，同时药物溶解度和口服生物利用度显著提高。The object of the present invention is to provide a kind of ibuprofen microemulsion delivery system, wherein the absolute dosage of surfactant is significantly reduced, and drug solubility and oral bioavailability are significantly improved simultaneously.
本发明提供一种布洛芬微乳给药系统，其包含表面活性剂、助表面活性剂、水、油和布洛芬，其中所述油为辛癸酸三甘油酯和三油酸甘油酯的混合油，质量比为6:1 - 1:6，所述辛癸酸三甘油酯中的辛酸部分和癸酸部分当分别形成辛酸甲酯和癸酸甲酯后，经气相色谱质谱联用仪测定的辛酸甲酯与癸酸甲酯的峰面积比为55:45~95:5。。在优选的实施方式中，所述辛癸酸三甘油酯与三油酸甘油酯的质量比为2:1 - 1:2，更优选为1:1。在一个实施方式中，辛酸甲酯与癸酸甲酯的峰面积比为75:25。The present invention provides a kind of ibuprofen microemulsion delivery system, it comprises surfactant, co-surfactant, water, oil and ibuprofen, wherein said oil is caprylic triglyceride and triolein Mixed oil, the mass ratio is 6:1 -1:6, after the caprylic acid part and the capric acid part in the described caprylic triglyceride are respectively formed caprylic acid methyl ester and capric acid methyl ester, the caprylic acid methyl ester and capric acid methyl ester that are measured by gas chromatography mass spectrometry instrument The peak area ratio is 55:45~95:5. . In a preferred embodiment, the mass ratio of caprylic capric triglyceride to triolein is 2:1-1:2, more preferably 1:1. In one embodiment, the peak area ratio of methyl caprylate to methyl caprate is 75:25.
在本发明的一个实施方式中，所述表面活性剂的含量是所述布洛芬含量的2-10倍。优选地，表面活性剂的含量仅为所述布洛芬含量的2倍。In one embodiment of the present invention, the content of the surfactant is 2-10 times that of the ibuprofen. Preferably, the content of the surfactant is only twice that of the ibuprofen.
在本发明的一个实施方式中，所述表面活性剂为质量比为2:1 - 1:2的吐温80和聚氧乙烯蓖麻油。优选地，吐温80与聚氧乙烯蓖麻油的质量比为1:1。In one embodiment of the present invention, the surfactant is Tween 80 and polyoxyethylene castor oil with a mass ratio of 2:1-1:2. Preferably, the mass ratio of Tween 80 to polyoxyethylene castor oil is 1:1.
在本发明的一个实施方式中，所述助表面活性剂选自聚乙二醇400、1,2-丙二醇、无水乙醇中的一种或多种。优选地，所述助表面活性剂为聚乙二醇400。In one embodiment of the present invention, the co-surfactant is selected from one or more of polyethylene glycol 400, 1,2-propylene glycol, and absolute ethanol. Preferably, the co-surfactant is polyethylene glycol 400.
在本发明的一个实施方式中，所述助表面活性剂与表面活性剂的质量比为1:1。In one embodiment of the present invention, the mass ratio of the co-surfactant to the surfactant is 1:1.
本发明另一方面提供所述布洛芬微乳给药系统的制备方法。本发明布洛芬微乳给药系统可通过常规的微乳制备方法制成。通常的制备方法包括：取处方量的表面活性剂和助表面活性剂，搅拌均匀；取处方量的混合油，用磁力搅拌器充分混合，与总表面活性剂混合，再加入处方量的药物（布洛芬），搅拌充分溶解，缓慢滴加处方量蒸馏水，即得澄清的微乳。Another aspect of the present invention provides a preparation method of the ibuprofen microemulsion drug delivery system. The ibuprofen microemulsion delivery system of the present invention can be prepared by conventional microemulsion preparation methods. Usual preparation method comprises: take the tensio-active agent of prescription quantity and co-surfactant, stir evenly; Take the mixed oil of prescription quantity, fully mix with magnetic stirrer, mix with total surfactant, then add the medicine of prescription quantity ( ibuprofen), stir to fully dissolve, and slowly add the prescribed amount of distilled water dropwise to obtain a clear microemulsion.
在该微乳系统中，使用中链油(即，辛癸酸三甘油酯 (GTCC))和不饱和脂肪酸酯(即，三油酸甘油酯 (Glycerol trioleate))作为混合油，吐温80 (Tween 80) 和聚氧乙烯蓖麻油 (Cremophor EL) 作为表面活性剂，聚乙二醇400 (PEG 400) 作为助表面活性剂。In this microemulsion system, medium-chain oil (i.e., caprylic triglyceride (GTCC)) and unsaturated fatty acid ester (i.e., triolein (Glycerol trioleate)) were used as mixed oil, Tween 80 (Tween 80) and polyoxyethylene castor oil (Cremophor EL) as surfactants, polyethylene glycol 400 (PEG 400) as co-surfactant.
与单一油相比较，本发明混合油的使用一方面显著扩大了微乳区域面积（MA为28.4 %，MA30 12.0 %），另一方面显著提高了布洛芬的溶解度（约60 mg/ml），两者的综合作用使制备载药微乳所需表面活性剂的绝对用量显著减少（例如，较单一油处方降低了六倍）。这意味着在载药量不变的情况下，制备载药微乳所需要的表面活性剂的绝对用量进一步降低。Compared with a single oil, the use of the mixed oil of the present invention significantly expands the area of the microemulsion area on the one hand (MA is 28.4%, MA30 12.0 %), on the other hand significantly improved the solubility of ibuprofen (about 60 mg/ml), the combined effect of the two significantly reduced the absolute amount of surfactant required for the preparation of drug-loaded microemulsions (for example, six times lower than single oil prescriptions). This means that under the condition of constant drug loading, the absolute amount of surfactant needed to prepare the drug-loading microemulsion is further reduced.
同时，与单一油微乳相比，本发明混合油的使用获得了相同的大鼠口服生物利用度，没有因表面活性剂用量较低而影响其提高药物口服生物利用度的能力，从而可以充分发挥药效，使布洛芬微乳广泛进入临床应用成为可能。Simultaneously, compared with single oil microemulsion, the use of mixed oil of the present invention has obtained identical rat oral bioavailability, does not have influence on its ability to improve drug oral bioavailability because of surfactant dosage is lower, thereby can fully Play drug effect, make ibuprofen microemulsion widely enter clinical application possible.
附图说明Description of drawings
图1为不同油相处方下绘制的伪三元相图及MA、MA30的柱状图，其中图1a是辛癸酸三甘油酯 (GTCC)作为油相；图1b是三油酸甘油酯 (Glycerol trioleate)作为油相；图1c是辛癸酸三甘油酯-三油酸甘油酯 (GTCC- Glycerol trioleate (1:1) )作为油相；图1d是图1a, 1b, 1c三幅伪三元相图的MA及MA30的柱状图；其中S-Cos表示总表面活性剂（即表面活性剂+助表面活性剂）。Fig. 1 is the pseudo-ternary phase diagram and the column diagram of MA and MA30 drawn under different oil phases, wherein Fig. 1a is triglyceride capricate (GTCC) as oil phase; Fig. 1b is glyceryl trioleate (Glycerol trioleate) as the oil phase; Figure 1c is caprylic capric triglyceride-triolein(GTCC- Glycerol trioleate (1:1) ) as the oil phase; Figure 1d is the histogram of MA and MA30 of the three pseudo-ternary phase diagrams of Figure 1a, 1b, 1c; where S-Cos represents the total surfactant (ie surfactant + co-surfactant).
图2显示布洛芬在辛癸酸三甘油酯 (GTCC)、油酸乙酯 (ethyl oleate)、单亚油酸甘油酯 (MaisineTM35-1)、三油酸甘油酯 (Glycerol trioleate)、肉豆蔻酸异丙酯 (IPM)、混合油 (GTCC-Glycerol trioleate，1:1、1:6、6:1)中的溶解度。与单一油相比，布洛芬在混合油的溶解度提高到258.5% - 310.0%。Figure 2 shows the activity of ibuprofen in triglyceride capricate (GTCC), ethyl oleate (ethyl oleate), glycerol monolinoleate (MaisineTM 35-1), glycerol trioleate (Glycerol trioleate), Solubility in isopropyl myristate (IPM), mixed oil (GTCC-Glycerol trioleate, 1:1, 1:6, 6:1). Compared with single oil, the solubility of ibuprofen in mixed oil increased to 258.5% - 310.0%.
图3显示布洛芬在不同微乳处方中的溶解度。其中ME-A为含24%总表面活性剂、6%单一油（辛癸酸三甘油酯）微乳；ME-B为含24%总表面活性剂、6%混合油(辛癸酸三甘油酯-三油酸甘油酯(1:1))微乳；ME-C为含20%总表面活性剂、12%混合油(辛癸酸三甘油酯-三油酸甘油酯(1:1))微乳；ME-X为含20%总表面活性剂、12%混合油(辛癸酸三甘油酯-三油酸甘油酯(6:1))微乳；ME-Y为含20%总表面活性剂、12%混合油(辛癸酸三甘油酯-三油酸甘油酯(2:1))微乳；ME-Z为含20%总表面活性剂、12%混合油(辛癸酸三甘油酯-三油酸甘油酯(1:2))微乳。Figure 3 shows the solubility of ibuprofen in different microemulsion formulations. Among them, ME-A is a microemulsion containing 24% total surfactant and 6% single oil (caprylic triglyceride); ME-B is a microemulsion containing 24% total surfactant and 6% mixed oil (caprylic triglyceride). ester-triolein (1:1)) microemulsion; ME-C contains 20% total surfactant, 12% mixed oil (caprylic triglyceride-triolein (1:1) ) microemulsion; ME-X is a microemulsion containing 20% total surfactant, 12% mixed oil (caprylic triglyceride-triolein (6:1)) microemulsion; ME-Y is a microemulsion containing 20% total Surfactant, 12% mixed oil (caprylic triglyceride-triolein (2:1)) microemulsion; ME-Z contains 20% total surfactant, 12% mixed oil (caprylic acid Triglyceride-triolein (1:2)) microemulsion.
图4显示布洛芬在不同微乳ME-A-Ibu (■)、ME-B-Ibu (●)、ME-C-Ibu (▲)、ME-D-Ibu (▼)、ME-E-Ibu (♦) 处方中37℃下的体外累积释放曲线。相同含量的布洛芬下，ME-E的表面活性剂用量最少，但体外累积释放曲线无显著性变化。Figure 4 shows that ibuprofen in different microemulsions ME-A-Ibu (■), ME-B-Ibu (●), ME-C-Ibu (▲), ME-D-Ibu (▼), ME-E- In vitro cumulative release profile of Ibu (♦) in formulation at 37°C. Under the same content of ibuprofen, ME-E uses the least amount of surfactant, but there is no significant change in the cumulative release curve in vitro.
图5显示大鼠口服不同微乳制剂及布洛芬混悬液 (25 mg/kg) 后的平均血药浓度-时间曲线(n = 5)。ME-A-Ibu (■), ME-B-Ibu (●), ME-C-Ibu (▲), ME-D-Ibu (▼), ME-E-Ibu (♦), 布洛芬混悬液(×)。Figure 5 shows that different microemulsion preparations and ibuprofen suspensions were orally administered to rats (25mg/kg) after the mean plasma concentration-time curve (n =5). ME-A-Ibu (■), ME-B-Ibu (●), ME-C-Ibu (▲), ME-D-Ibu (▼), ME-E-Ibu (♦), Ibuprofen Suspension Liquid (×).
具体实施方式Detailed ways
本发明采用生物药剂学分类系统II类（BCS class II）药物布洛芬作为模型药物，采用中链油辛癸酸三甘油酯和三油酸甘油酯的混合油为油相，制备微乳给药系统。研究发现，该混合油不仅能够显著扩大微乳区域面积，并且能够明显提高布洛芬的溶解度。与单一油微乳相比，优化的混合油微乳具有相当的大鼠口服生物利用度，而其表面活性剂用量仅为单一油微乳中的六分之一。以下通过实验进一步证实本发明的效果。在本发明的伪三元相图和微乳给药系统的制备以及布洛芬溶解度的测定均在25℃的条件下进行。辛癸酸三甘油酯是辛酸、癸酸与甘油酯化形成的酯的混合物，可购自成都格雷西亚化学技术有限公司（产品编号：1022224；批号：I4285）。The present invention adopts biopharmaceutical classification system II (BCS class II) drug ibuprofen as a model drug, and uses a mixed oil of medium-chain oil caprylic triglyceride and triolein as the oil phase to prepare microemulsion for drug delivery. medicine system. The study found that the mixed oil can not only significantly expand the area of the microemulsion, but also significantly improve the solubility of ibuprofen. Compared with single oil microemulsion, the optimized mixed oil microemulsion has comparable oral bioavailability in rats, while its surfactant dosage is only one-sixth of that of single oil microemulsion. The effects of the present invention are further confirmed by experiments below. The preparation of the pseudo-ternary phase diagram and the microemulsion drug delivery system of the present invention and the determination of the solubility of ibuprofen are all carried out under the condition of 25°C. Caprylic capric triglyceride is a mixture of esters formed by the esterification of caprylic acid, capric acid and glycerol, which can be purchased from Chengdu Gracia Chemical Technology Co., Ltd. (product number: 1022224; batch number: I4285).
实施例Example11微乳伪三元相图的制备Preparation of Pseudo-ternary Phase Diagram of Microemulsion
称取一定量的表面活性剂和助表面活性剂，用磁力搅拌器充分混合1小时，即得总表面活性剂S-Cos。在25℃下，分别称取S-Cos 1.8、1.6、1.4、1.2、1.0、0.8、0.6、0.4、0.2 g于西林瓶中，再分别加入0.2、0.4、0.6、0.8、1.0、1.2、1.4、1.6、1.8 g油相（a的油相为辛癸酸三甘油酯，b的油相为三油酸甘油酯，c的油相为辛癸酸三甘油酯-三油酸甘油酯（1:1）），混匀，在磁力搅拌下向混合液中缓慢滴加蒸馏水，以外观“澄清透明”为指标，记录加入水量，按油、总表面活性剂、水在临界点的各自质量百分数（w/w），绘制伪三元相图，确定微乳区，用黑色区域表示，结果见图1a、1b和1c。 Weigh a certain amount of surfactant and co-surfactant, and mix thoroughly with a magnetic stirrer for 1 hour to obtain the total surfactant S-Cos. At 25°C, weigh S-Cos 1.8, 1.6, 1.4, 1.2, 1.0, 0.8, 0.6, 0.4, 0.2 g into vials, then add 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4 , 1.6, 1.8 g oil phase (the oil phase of a is caprylic triglyceride, the oil phase of b is triolein, the oil phase of c is caprylic triglyceride-triolein (1 :1)), mix well, slowly add distilled water dropwise to the mixture under magnetic stirring, take the appearance of "clear and transparent" as an indicator, record the amount of water added, according to the respective mass percentages of oil, total surfactant, and water at the critical point (w/w), draw a pseudo-ternary phase diagram, and determine the microemulsion area, which is represented by the black area. The results are shown in Figures 1a, 1b, and 1c.
微乳区域面积占伪三元相图三角形面积的百分比（percentage of microemulsion area，MA）及总表面活性剂用量小于30 %时微乳区域占伪三元相图三角形面积的百分比（MA30）。根据伪三元相图，用计算机软件计算得具体数值，绘制柱状图，结果见图1d。可见，混合油微乳的MA及MA30均明显比单一油微乳的MA及MA30大。The percentage of microemulsion area in the triangle area of the pseudo-ternary phase diagram (percentage of microemulsion area, MA) and the percentage of microemulsion area in the triangle area of the pseudo-ternary phase diagram when the total surfactant dosage is less than 30% (MA30). According to the pseudo-ternary phase diagram, the specific values were calculated by computer software, and the histogram was drawn. The results are shown in Figure 1d. It can be seen that the MA and MA30 of the mixed oil microemulsion are significantly larger than the MA and MA30 of the single oil microemulsion.
实施例Example22布洛芬ibuprofenHPLCHPLC含量测定方法的建立Establishment of content determination method
色谱条件：色谱柱为GraceSmart C18(250×4.6 mm，5 μm)，流动相为甲醇-pH3.0磷酸盐缓冲液 (60:40) ，流速1 mL/min，进样量20μL，检测波长λ = 220 nm，柱温35oC。Chromatographic conditions: GraceSmart C18 column (250×4.6 mm, 5 μm), mobile phase methanol-pH3.0 phosphate buffer (60:40), flow rate 1 mL/min, injection volume 20 μL, detection wavelength λ = 220 nm, column temperature 35o C.
标准曲线的建立：以甲醇为溶剂，配制浓度为40、20、10、5、2.5、1.2、0.6 μg/mL的布洛芬系列溶液，按所定的色谱条件分别进样分析，以布洛芬峰面积对浓度作标准曲线。可知，布洛芬在0.6-40 μg/mL范围内，峰面积与浓度线性相关性良好 (A=57.103C-5.6333, R2=0.9997)。Establishment of the standard curve: using methanol as a solvent, prepare a series of ibuprofen solutions with concentrations of 40, 20, 10, 5, 2.5, 1.2, and 0.6 μg/mL, and inject and analyze them respectively according to the predetermined chromatographic conditions. The peak area versus concentration was used as a standard curve. It can be seen that within the range of 0.6-40 μg/mL of ibuprofen, the peak area has a good linear correlation with the concentration (A=57.103C-5.6333, R2 =0.9997).
实施例Example33布洛芬在水中的溶解度Solubility of ibuprofen in water
取蒸馏水2 mL于西林瓶中，平行3份，加入适量过量布洛芬至出现沉淀，涡旋3~5 min，促进蒸馏水溶解药物，封口。置于25oC恒温水浴摇床中，100 rpm平衡72 h。15000 rpm离心5 min，取上清液，用甲醇经适当稀释后，HPLC法测定药物含量，即得到布洛芬在水中的溶解度约为0.07 mg/mL。Take 2 mL of distilled water in a vial, make 3 parallel portions, add an appropriate amount of ibuprofen until precipitation occurs, vortex for 3-5 min to promote the dissolution of the drug in distilled water, and seal it. Placed in a constant temperature water bath shaker at 25o C, and equilibrated at 100 rpm for 72 h. After centrifugation at 15000 rpm for 5 min, the supernatant was taken, diluted appropriately with methanol, and the drug content was determined by HPLC. The solubility of ibuprofen in water was about 0.07 mg/mL.
实施例Example44布洛芬在不同油相中的溶解度Solubility of ibuprofen in different oil phases
取油2 mL于西林瓶中，平行3份，加入适量过量布洛芬至出现沉淀，涡旋3~5 min，促进油相溶解药物，封口。置于25 oC恒温水浴摇床中，100 rpm平衡72 h。15000 rpm离心5 min，取上清液，用甲醇经适当稀释后，HPLC法测定药物含量，即得布洛芬在不同油相中的溶解度。结果如图2所示(辛癸酸三甘油酯: 96.15 mg/mL; 三油酸甘油酯: 115.30 mg/mL; 辛癸酸三甘油酯-三油酸甘油酯 (1:1) : 298.07 mg/mL；辛癸酸三甘油酯-三油酸甘油酯 (6:1) : 285.30 mg/mL；辛癸酸三甘油酯-三油酸甘油酯 (1:6) : 270.15 mg/mL)。Take 2 mL of the oil in a vial, make 3 parallel portions, add an appropriate amount of ibuprofen until precipitation occurs, vortex for 3-5 min to promote the dissolution of the drug in the oil phase, and seal it. Place in a constant temperature water bath shaker at 25oC , and equilibrate at 100 rpm for 72 h. Centrifuge at 15,000 rpm for 5 min, take the supernatant, dilute it properly with methanol, and measure the drug content by HPLC to obtain the solubility of ibuprofen in different oil phases. The results are shown in Figure 2 (caprylic triglyceride: 96.15 mg/mL; triolein: 115.30 mg/mL; caprylic triglyceride-triolein (1:1): 298.07 mg /mL; caprylic triglyceride-triolein (6:1): 285.30 mg/mL; caprylic triglyceride-triolein (1:6): 270.15 mg/mL).
实施例Example55微乳体系的制备Preparation of microemulsion system
在25℃下，根据表1称取处方量的表面活性剂和助表面活性剂，用磁力搅拌器充分混合，即得总表面活性剂。另取处方量油相，混合均匀，得混合油。混合油再与总表面活性剂混合，在搅拌下缓慢滴加处方量水，即得空白微乳ME-A、ME-B、ME-C、ME-F、ME-G、ME-H、ME-I、ME-J、ME-X、ME-Y、ME-Z。At 25°C, weigh the prescribed amount of surfactant and co-surfactant according to Table 1, and mix thoroughly with a magnetic stirrer to obtain the total surfactant. Separately take the prescribed amount of oil phase, and mix evenly to obtain a mixed oil. Mix the mixed oil with the total surfactant, and slowly add the prescribed amount of water dropwise under stirring to obtain blank microemulsions ME-A, ME-B, ME-C, ME-F, ME-G, ME-H, ME -I, ME-J, ME-X, ME-Y, ME-Z.
表1微乳的处方组成The prescription composition of table 1 microemulsion
注：总表面活性剂：吐温80-聚氧乙烯蓖麻油-聚乙二醇400 (1:1:2)Note: Total surfactant: Tween 80-polyoxyethylene castor oil-polyethylene glycol 400 (1:1:2)
其中中括号[]表示所述辛癸酸三甘油酯中的辛酸部分和癸酸部分当分别形成辛酸甲酯和癸酸甲酯后，经气相色谱质谱联用（GC-MS）仪测定的辛酸甲酯与癸酸甲酯的峰面积比。确定所述峰面积比的程序如下：取1滴样品加入2 mL 2%氢氧化钾甲醇溶液于60 ℃水浴反应15 min，正己烷萃取上清液，用10 mL水水洗，取有机层用无水硫酸钠脱水进样。Wherein the square brackets [ ] represent the octanoic acid part and the capric acid part in the caprylic capric acid triglyceride. Peak area ratio of methyl ester to methyl caprate. The procedure for determining the peak area ratio is as follows: take 1 drop of sample and add 2 mL of 2% potassium hydroxide methanol solution to react in a water bath at 60 °C for 15 min, extract the supernatant with n-hexane, wash with 10 mL of water, take the organic layer and wash it with Sodium sulfate dehydration injection.
仪器名称：气相色谱质谱联用仪（GC-MS）Instrument Name: Gas Chromatography Mass Spectrometry (GC-MS)
厂家：THERMOL FINNIGANManufacturer: THERMOL FINNIGAN
型号：TRACE DSQModel: TRACE DSQ
柱子：DB-5MSColumn: DB-5MS
进样口温度：230 ℃Injection port temperature: 230 ℃
进样体积：1mLInjection volume: 1mL
柱温箱温度：190 ℃（恒温，保持34 min）Column oven temperature: 190 °C (constant temperature, keep for 34 min)
离子源温度：230 ℃Ion source temperature: 230 ℃
溶剂延迟时间：3 minSolvent delay time: 3 min
分流比：30:1Split ratio: 30:1
气相-质谱连接处温度：230 ℃。Gas phase-mass spectrometry junction temperature: 230°C.
实施例Example66布洛芬在不同微乳处方中的溶解度Solubility of ibuprofen in different microemulsion formulations
取ME-A、ME-B、ME-C、ME-X、ME-Y、ME-Z空白微乳各2 mL于西林瓶中，平行3份，分别加入适量过量药物，直至出现沉淀，涡旋3~5min，促进药物溶解，封口。再置于25℃恒温水浴摇床中，100 rpm平衡24 h。15000 rpm离心5 min，取上清液，用甲醇稀释后，HPLC法测定药物含量。布洛芬在不同微乳处方中的溶解度如图3所示(ME-A: 15.30 mg/mL; ME-B: 31.71 mg/mL; ME-C: 59.59 mg/mL；ME-X: 55.25 mg/mL；ME-Y: 57.95 mg/mL；ME-Z: 56.25 mg/mL)。Take 2 mL each of ME-A, ME-B, ME-C, ME-X, ME-Y, and ME-Z blank microemulsions in a vial, and add an appropriate amount of excess drug in parallel to 3 portions, until precipitation occurs, vortex Rotate for 3-5 minutes to promote drug dissolution and seal. Then place it in a constant temperature water bath shaker at 25°C, and equilibrate at 100 rpm for 24 h. Centrifuge at 15000 rpm for 5 min, take the supernatant, dilute with methanol, and determine the drug content by HPLC. The solubility of ibuprofen in different microemulsion formulations is shown in Figure 3 (ME-A:15.30 mg/mL; ME-B: 31.71 mg/mL;ME-C: 59.59 mg/mL; ME-X:55.25 mg/mL; ME-Y:57.95 mg/mL; ME-Z:56.25 mg/mL).
实施例Example77不同处方的布洛芬微乳在体外的释放实验In Vitro Release Experiment of Ibuprofen Microemulsions with Different Prescriptions
根据表2，称取处方量的表面活性剂加入助表面活性剂，用磁力搅拌器充分混合1小时，即得总表面活性剂。取处方量油相混合均匀，得混合油。在25℃下，将混合油与总表面活性剂混合，再加入处方量布洛芬，搅拌充分溶解，在搅拌下缓慢滴加处方量蒸馏水，即得澄清的布洛芬微乳ME-A-Ibu、ME-B-Ibu、ME-C-Ibu、ME-D-Ibu、ME-E-Ibu。According to Table 2, the surfactant of the recipe amount was weighed and added into the co-surfactant, and thoroughly mixed with a magnetic stirrer for 1 hour to obtain the total surfactant. Take the prescribed amount of oil and mix evenly to obtain a mixed oil. At 25°C, mix the mixed oil with the total surfactant, then add the prescribed amount of ibuprofen, stir to fully dissolve, and slowly add the prescribed amount of distilled water dropwise under stirring to obtain a clear ibuprofen microemulsion ME-A- Ibu, ME-B-Ibu, ME-C-Ibu, ME-D-Ibu, ME-E-Ibu.
取各微乳制剂各1mL置于透析袋 (MW = 3500kDa) 中，将其分别置于50mL的PBS溶液中；将其分别置于恒温震荡器中，水浴37 oC震荡，100 rpm，并分别于0.5、1、1.5、2、3、4、5、6、8、16、24 h吸取1mL释放介质，并及时补充释放介质，HPLC法测定药物含量，并绘制体外累积释放曲线，如图4所示。Take each 1mL of each microemulsion preparation and place it in a dialysis bag (MW = 3500kDa), place it in 50mL of PBS solution respectively; place it in a constant temperature shaker, shake in a water bath at 37oC , 100 rpm, and place in a At 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, 16, and 24 hours, 1 mL of release medium was drawn, and the release medium was replenished in time. The drug content was determined by HPLC, and the cumulative release curve in vitro was drawn, as shown in Figure 4. Show.
表2 布洛芬微乳的处方组成Table 2 The prescription composition of ibuprofen microemulsion
注：Note:
a 辛癸酸三甘油酯中的辛酸部分和癸酸部分分别形成辛酸甲酯和癸酸甲酯后，经气相色谱质谱联用（GC-MS）仪测定的辛酸甲酯与癸酸甲酯的峰面积比为55:45~95:5a After the caprylic acid part and capric acid part in caprylic triglyceride form caprylic acid methyl ester and capric acid methyl ester respectively, the ratio of caprylic acid methyl ester and capric acid methyl ester determined by gas chromatography-mass spectrometry (GC-MS) The peak area ratio is 55:45~95:5
b总表面活性剂：吐温80-聚氧乙烯蓖麻油-聚乙二醇400 (1:1:2)b Total surfactant: Tween 80-polyoxyethylene castor oil-polyethylene glycol 400 (1:1:2)
c 载药微乳中每10 mg布洛芬所对应的吐温80的用量。c The amount of Tween 80 per 10 mg ibuprofen in the drug-loaded microemulsion.
实施例Example88不同处方的布洛芬微乳在大鼠体内的生物利用度实验Experiment of bioavailability of ibuprofen microemulsions with different prescriptions in rats
以布洛芬混悬剂为参比，分别口服灌胃布洛芬混悬剂及实施例7所制备的各微乳，考察布洛芬混合油微乳、单一油微乳对布洛芬生物利用的提高，考察混合油与单一油微乳生物利用度的差异，及表面活性剂用量下降是否影响药物生物利用度的提高。Taking ibuprofen suspension as reference, each microemulsion prepared by oral gavage ibuprofen suspension and embodiment 7 respectively investigates the effect of ibuprofen mixed oil microemulsion and single oil microemulsion on ibuprofen biological To improve the utilization, investigate the difference in the bioavailability of mixed oil and single oil microemulsion, and whether the decrease of surfactant dosage affects the improvement of drug bioavailability.
取雄性SD大鼠30只，体重200±20 g，随机分为6组，每组5只动物。按布洛芬25 mg/kg的剂量，对SD大鼠口服灌胃各处方制剂。给药前12 h禁食不禁水，给药后4 h自由进食，采集血样过程都给予自由饮水，并补充生理盐水3次。Thirty male SD rats, weighing 200±20 g, were randomly divided into 6 groups, with 5 animals in each group. According to the dose of ibuprofen 25 mg/kg, SD rats were orally gavaged with each prescription preparation. They were fasted for 12 hours before the administration, and had free access to food 4 hours after the administration. They were given free drinking water during blood sample collection, and supplemented with normal saline 3 times.
口服灌胃给药后分别于5、10、20、30、45、60、90、120、240、360、480 min用采血针取血。取血浆50 μL加入50 μL乙腈，再加入788.8 ng/mL的内标萘普生溶液150 μL，涡旋30 s，15 ℃ 13000 rpm离心10 min。取上清血浆100 μL加入900 μL蒸馏水，15 ℃ 13000 rpm离心10 min，取上清液，用LC-MS/MS方法对样品进行处理及测定。将计算结果绘制成平均血药浓度-时间曲线如图5所示。相关的药代动力学参数如表3所示。Blood was collected with a blood collection needle at 5, 10, 20, 30, 45, 60, 90, 120, 240, 360, and 480 min after oral gavage administration. Take 50 μL of plasma and add 50 μL of acetonitrile, then add 150 μL of 788.8 ng/mL internal standard naproxen solution, vortex for 30 s, and centrifuge at 13000 rpm at 15 °C for 10 min. Take 100 μL supernatant plasma and add 900 μL distilled water, centrifuge at 13000 rpm at 15 °C for 10min, take the supernatant, and use the LC-MS/MS method to process and measure the sample. The calculated results were plotted into the average blood drug concentration-time curve as shown in Figure 5. The relevant pharmacokinetic parameters are shown in Table 3.
表3 大鼠口服不同微乳制剂的相关药代动力学参数Table 3 Pharmacokinetic parameters related to oral administration of different microemulsion preparations in rats
注：与布洛芬混悬剂组比较，*P<0.05;**P<0.01。Note: Compared with the ibuprofen suspension group,* P<0.05;** P<0.01.
本发明通过处方优化，选用中链饱和油和长链不饱和油制备的混合油微乳，与单一油微乳相比，布洛芬混合油微乳的表面活性剂用量仅为单一油的六分之一，表面活性剂用量最优时仅为布洛芬的2倍。The present invention selects mixed oil microemulsions prepared from medium-chain saturated oils and long-chain unsaturated oils through prescription optimization. Compared with single oil microemulsions, the surfactant dosage of ibuprofen mixed oil microemulsions is only six times that of single oils. One-third, the optimal amount of surfactant is only 2 times that of ibuprofen.
大鼠口服生物利用度结果表明，虽然混合油微乳与单一油微乳表面活性剂绝对用量相差10倍，但口服生物利用度相当。综上，本发明采用混合油，不仅扩大了微乳区域面积，而且同时提高了药物的溶解度，从而显著降低了制备载药微乳所需的表面活性剂的绝对用量，降低了由于表面活性剂用量大导致的微乳的毒副作用，为微乳产业化提供了可能的途径。The results of oral bioavailability in rats showed that although the absolute dosage of surfactants in mixed oil microemulsion and single oil microemulsion differed by 10 times, the oral bioavailability was equivalent. In summary, the present invention adopts mixed oil, not only expands the area of microemulsion area, but also improves the solubility of medicine simultaneously, thus significantly reduces the absolute consumption of the surfactant required for preparing drug-loaded microemulsion, reduces the amount due to surfactant The toxic and side effects of microemulsions caused by large dosage provide a possible way for the industrialization of microemulsions.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510162641.1ACN104800159A (en) | 2015-04-08 | 2015-04-08 | Ibuprofen microemulsion drug delivery system |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510162641.1ACN104800159A (en) | 2015-04-08 | 2015-04-08 | Ibuprofen microemulsion drug delivery system |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201510162641.1APendingCN104800159A (en) | 2015-04-08 | 2015-04-08 | Ibuprofen microemulsion drug delivery system |
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| CN (1) | CN104800159A (en) |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992008445A1 (en)* | 1990-11-13 | 1992-05-29 | Affinity Biotech, Inc. | Non-aqueous microemulsion for drug delivery |
| CN101904815A (en)* | 2009-06-03 | 2010-12-08 | 河北奥星集团药业有限公司 | A kind of ibuprofen microemulsion preparation and preparation method |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992008445A1 (en)* | 1990-11-13 | 1992-05-29 | Affinity Biotech, Inc. | Non-aqueous microemulsion for drug delivery |
| CN101904815A (en)* | 2009-06-03 | 2010-12-08 | 河北奥星集团药业有限公司 | A kind of ibuprofen microemulsion preparation and preparation method |
| Title |
|---|
| XIUHUA YOU ET AL.: "Optimizing surfactant content to improve oral bioavailability of ibuprofen in microemulsions: Just enough or more than enough?", 《INTERNATIONAL JOURNAL OF PHARMACEUTICS》* |
| YIZHEN CHEN ET AL.: "Optimized mixed oils remarkab...enhancing drug solubility", 《INTERNATIONAL JOURNAL OF PHARMACEUTICS》* |
| 陈华兵等: "布洛芬微乳的制备及其透皮吸收研究", 《中国药学杂志》* |
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