WO2014026502A1 - 蚕蛹油在提高小分子药物生物利用度中的用途 - Google Patents

蚕蛹油在提高小分子药物生物利用度中的用途 Download PDF

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WO2014026502A1
WO2014026502A1 PCT/CN2013/077513 CN2013077513W WO2014026502A1 WO 2014026502 A1 WO2014026502 A1 WO 2014026502A1 CN 2013077513 W CN2013077513 W CN 2013077513W WO 2014026502 A1 WO2014026502 A1 WO 2014026502A1
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small molecule
puerarin
drug
bioavailability
oil
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French (fr)
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张耀洲
张丽
吴少峰
陈剑清
舒特俊
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天津耀宇生物技术有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives

Definitions

  • the present invention relates to the use of silkworm pupa oil for improving the bioavailability of small molecule drugs, and belongs to the field of biomedicine. Background technique
  • the preparation of the dosage form Before any application of the drug substance to the clinic, it must be made into a certain dosage form.
  • the preparation of the dosage form also has some auxiliary materials to facilitate the stabilization, solubilization, solubilization, sustained release, etc. of the drug.
  • these auxiliary materials are collectively referred to as pharmaceutical excipients or pharmaceutical excipients.
  • the development of a pharmaceutical excipient is far more important than the development of a new drug.
  • the selection of pharmaceutical excipients is appropriate, which will directly affect the bioavailability, side effects and adverse reactions of the drugs.
  • Cardiovascular and cerebrovascular diseases are one of the diseases that endanger human life and health. Flavonoids have been shown to be effective in the treatment of cardiovascular and cerebrovascular diseases and are safe to use. Puerarin is a very representative one of flavonoids. Its pharmacological aspects have been widely used in cardiovascular and cerebrovascular diseases such as arteriosclerosis, hypertension, coronary insufficiency, angina pectoris, myocardial infarction, hyperlipidemia, cerebral infarction, cerebral vasospasm, senile dementia prevention and other diseases. Anti-thrombotic, microcirculation, and antioxidant effects are also evident in pharmacological action.
  • the present invention seeks to improve the bioavailability of the small molecule drug puerarin, and to detect the bioavailability of puerarin.
  • Puerarin preparations such as puerarin injection or puerarin preparations have great disadvantages, such as injection, because puerarin is a small molecule drug extracted from Chinese medicine Pueraria, not in the process of extraction, purification, etc.
  • the purity of one hundred, the trace impurities contained in it are easy to cause toxic side effects after injection, such as allergic hemolysis and other adverse reactions.
  • the oral absorption is very low due to the unique water insolubility of puerarin. The curative effect is not good and the effect is slow.
  • the silkworm pupa oil as an auxiliary material has nearly 5 times higher bioavailability in oral administration, and the silkworm pupa oil has no toxic side effects, which is a good health care itself. food. And the operation is simple, and the mixture can be administered. Summary of the invention
  • the object of the present invention is to provide a use of silkworm pupa oil for improving the bioavailability of small molecule drugs, in particular, the use of silkworm pupa oil as a medicinal adjuvant in improving the bioavailability of small molecule drugs.
  • one aspect of the present invention relates to the use of silkworm pupa oil for improving the bioavailability of small molecule drugs.
  • Another aspect of the invention relates to the use of silkworm pupa as a pharmaceutical excipient for increasing the bioavailability of small molecule drugs.
  • the small molecule drug is administered to the subject in a form mixed with silkworm pupa oil.
  • the small molecule drug is mixed with silkworm pupa oil at a ratio of 10 mg/ml, and ground for 10 min, sonicated for 30 min, and thoroughly mixed to obtain a mixture.
  • each dose of the mixture administered to the subject is 0.2 ml/10 g body weight based on the mixture of the drug and the silkworm pupa oil.
  • the mixture is administered to the subject by gavage.
  • the subject is a mouse.
  • the small molecule drug is puerarin.
  • the invention adopts different silkworm cocoons and puerarin to be intragastrically mixed, and the blood concentration of the mice is detected by high performance liquid chromatography. The blood is taken through the 30 min time point to find the components for increasing the blood concentration of puerarin. It was found that silkworm pupa oil had the strongest improvement ability; silkworm pupa oil as a medicinal auxiliary material could increase the relative bioavailability of puerarin by 4.7 times and increase the drug absorption concentration by 6-9 times at 30 min. Moreover, the cocoon oil safety Stable, no adverse effects, and extremely broad application prospects. DRAWINGS
  • Figure 1 is a blank serum chromatogram provided in the present invention
  • Figure 2 shows the chromatogram of puerarin and 4-hydroxyformaldehyde added to blank serum:
  • Figure 3 is a chromatogram of a plasma sample
  • Figure 4 shows the intention of puerarin blood concentration after treatment of each component
  • Figure 5 is a blood drug-time graph. detailed description
  • Example 1 Exploration of liquid phase conditions and selection of internal standards
  • mice After taking the blood, the mice were allowed to stand at room temperature for 30 min, centrifuged at 3500 rpm for 3 min, and the supernatant was taken 190 ⁇ l in a 1.5 ml EP tube. ⁇ , 4-hydroxybenzaldehyde internal standard solution (100 g/ml) was added. Shake and mix well, force into 400 ⁇ 1 acetonitrile, vortex for 5min, centrifuge at 12000rpm for 10min, take the supernatant after 0.22 ⁇ organic membrane, and load 20 ⁇ 1.
  • Calculation of absolute recovery rate Take 190 ⁇ 1 of mouse blank serum and place it in three sputum tubes. Add 10, 50, 100 g/ml puerarin methanol standard solution, which is high, medium and low, and take the same amount. Two copies of the drug standard and the internal standard substance, one part is added to the serum, and the other part is added to the mobile phase solvent, and the ratio of the drug to the internal standard peak area is measured. The measured results are shown in Table 2 and Table 3 below.
  • Precision includes intra-day precision and day-to-day precision. Intra-day precision measures the sample three times on the same day, and the daytime precision measures the sample three times at different times. Take 190 ⁇ l of mouse blank serum and place it in three EP tubes. Add puerarin methanol solution to obtain three low, medium and high concentrations and measure according to the method of "Separation 2 serum treatment and determination". The samples will be low, medium and high on the same day. The intraday precision obtained by three times of the results was measured, and the treated sample was measured three times a day for three days, and the obtained result was daytime precision, and the results are shown in Table 4 and Table 5 below.
  • the amount of addition (ng/ml) is high 2000 2000 2000 2000 2000
  • the mixture was treated with water and puerarin as the control group.
  • the treatment group was fresh silkworm blood group, simulated liposome group, freeze-dried silkworm pupa group and silkworm pupa oil group.
  • the concentration of control group, fresh silkworm blood group and silkworm pupa group Both are 10mg/ml, that is, 10mg puerarin (Netizen Chemical (Jinan) Co., Ltd.) is dissolved in 1ml of solvent ((water, fresh silkworm blood (take 50g silkworm cocoon grinding and crushing, centrifuge at 500rpm for 1Omin and take the supernatant) , silkworm pupa oil (National Grain Reserve Bureau Xi'an Research Institute of Oil Science and Technology)); simulated liposome group, fresh silkworm blood, absolute ethanol, sodium carboxymethyl cellulose and water (fresh silkworm blood: absolute ethanol : 0.5% sodium carboxymethyl cellulose solution: water volume ratio of 2: 1 : 4: 0.5) mixed into a solution, then puerarin is dissolved
  • the amount is 0.2ml/10g (each time), blood is taken 30min after each groping component is administered, the test result is shown in Figure 4, looking for Best component chrysalis oil, oil mixed in pupa puerarin gavage excipients and 5min, 10min, 20min, 3 Omin, 40min, Six mice were bled at lh, 1.5h, 2h, 2.5h, 3h, 4h, 6h, 8h, and 1 Oh.
  • the blood sample treatment method is the same as the standard curve, determined by high performance liquid chromatography, and the measured area is brought to the concentration obtained by the standard curve.
  • Example 8 Calculation of various parameters and bioavailability multiples using 3p89 software The blood drug concentrations measured are shown in Figure 5, and the data is shown in Table 9 below.
  • Tmax drug peak time
  • Cmax drug peak concentration
  • AUC area under the blood drug concentration curve
  • Ke apparent first-order absorption rate constant
  • T1/2 half-life
  • AUCT and AUCR are taken respectively
  • the area under the blood drug curve (control group), DT and DR are the doses of the test preparation and the reference preparation, respectively (the doses here are all 10 mg/ml, 0.2 ml/10 g).

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Abstract

本发明涉及蚕蛹油在提高小分子药物生物利用度中的用途，属于生物医药领域。本发明主用利用家蚕为材料，寻找到能够有效促进小分子药物（葛根素）的生物利用的蚕蛹组分，即蚕蛹油，并对其进行了验证和计算，最终发现与对照相比提高了4.7倍，就30min时间点来说就高于对照6-9倍之多；该蚕蛹油安全稳定，无副毒作用，应用前景极其广泛。

Description

蚕蛹油在提高小分子药物生物利用度中的用途 技术领域 本发明涉及蚕蛹油在提高小分子药物生物利用度中的用途，属于 生物医药领域。 背景技术
任何一种原料药在应用到了临床之前， 都必须制成一定的剂型， 剂型的形成除了这些原料药外还要有一些辅助材料从而有利于这些 药的稳定、 增溶、 助溶、 缓释等,通常这些辅助材料统称为药用辅料 或药物辅料。一种药用辅料的开发，其意义要远超过一种新药的开发。 在药物制剂的制备过程中， 药用辅料的选用是否得当， 将直接影响药 物的生物利用度、 毒副作用、 不良反应。
我国的药用辅料品种少， 新型辅料研究应用相对滞后， 并且对于 大多数药物， 口服给药是最好的给药选择， 以提高病人的依附性， 且 经济、安全。如何解决难溶性药物的口服吸收问题， 已成为一个热点。
心脑血管疾病是危害人类生命和健康的疾病之一。黄酮类化合物 被证实在治疗心脑血管疾病方面疗效肯定， 使用安全性较高。 葛根素 是黄酮类化合物中极具代表性的一种。其药理方面已广泛应用于心脑 血管疾病如动脉硬化、 高血压、 冠脉供血不全、 心绞痛、 心肌梗死、 高脂血症、 脑梗死、 脑血管痉挛、 老年性痴呆防治等疾病。 在药理作 用中其抗血栓， 改善微循环， 以及抗氧化作用等也很明显。
基于以上内容，本发明皆在寻找能够提高小分子药物葛根素生物 利用度的组分， 并对葛根素的生物利用度进行检测。 对于巿面上销售 的葛根素制剂如葛根素注射液或是含葛根素制剂均存在很大的不足， 比如注射液，因为葛根素是从中药葛根中提取的小分子药物，在提取、 纯化等过程中不能百分百的纯度，所含的微量杂质在注射后很容易造 成毒副作用， 比如过敏溶血等不良反应， 对于能够上巿的片剂来说， 由于葛根素独特的水不溶性， 其口服吸收很低， 从而疗效不佳且效果 也比较慢， 这些都是其不如临床的局限， 而蚕蛹油作为辅料， 在口服 中其生物利用度提高了将近 5倍， 并且蚕蛹油无毒副作用， 本身就是 很好的保健食品。 并且操作简单， 混合灌胃即可。 发明内容
有鉴于此，本发明的目的在于提供蚕蛹油在提高小分子药物生物 利用度中的用途，尤其是蚕蛹油作为药用辅料在提高小分子药物生物 利用度中的用途。
为达到上述目的，本发明一方面涉及蚕蛹油在提高小分子药物生 物利用度中的用途。
本发明另一方面涉及蚕蛹油作为药用辅料在提高小分子药物生 物利用度中的用途。
优选地， 其中所述小分子药物以与蚕蛹油混合的形式给予受试 者。
优选地， 其中所述小分子药物与蚕蛹油以 10mg/ml的比例混合， 并研磨 lOmin, 超声 30min, 充分混合而得到混合物。
优选地，其中所述混合物给予受试者的每曰剂量以药物与蚕蛹油 的混合物计为 0.2ml/10g体重。
优选地， 其中所述混合物通过灌胃的方式给予受试者。
优选地， 其中所述受试者为小鼠。
优选地， 其中所述小分子药物为葛根素。 本发明釆用不同的蚕蛹组分与葛根素混合灌胃，用高效液相色谱 检测小鼠体内血药浓度， 先通过 30min时间点取血检测， 寻找到提高 葛根素血药浓度的组分， 最终发现蚕蛹油的提高能力最为强； 蚕蛹油 作为药用辅料可使葛根素的相对生物利用度提高 4.7倍， 在 30min时 的药物吸收浓度提高 6-9倍之多； 而且， 该蚕蛹油安全稳定， 无副毒 作用， 应用前景极其广泛。 附图说明
图 1为本发明中提供的空白血清色谱图
图 2为空白血清中加入葛根素和 4-羟基 甲醛色谱图:
图 3为血浆样品色谱图；
图 4为各组分处理后葛根素血药浓度的 意图;
图 5为血药-时间曲线图。 具体实施方式
应该指出， 以下具体说明都是例示性的， 皆在对本发明提供进一 步的发明。 除非另有说明， 本文使用的所有科学和技术术语具有与本 发明所属技术领域人员通常理解的相同含义。
下面结合实施例详细阐述本发明的具体内容。
实施例 1 : 液相条件的摸索以及内标的选择
色谱柱 Venusil XBP C 18(4.6mm 250mm,5m); 流动相为甲醇： 水 (30:70)(v/v); 流速 0.7ml/min, 柱温为 30°C ; 紫外检测波长 250nm; 进样量为 201内标为 4-羟基苯甲醛。
实施例 2: 血清处理与测定
小鼠取血后室温静止 30min, 4°C 3500rpm离心 lOmin, 取上清 190μ1于 1.5mlEP管中，加入 ΙΟμΙ, 4-羟基苯甲醛内标溶液（lOO g/ml) 振摇混匀， 力口入 400μ1乙腈， 涡旋振荡 5min, 12000rpm 离心 lOmin, 取上清过 0.22μηι有机膜后， 上样 20μ1。
实施例 3 : 标准储备液以及标准曲线的制备
精密称取葛根素标准品 10mg, 置于 10ml容量瓶中，加甲醇溶解 并稀释至刻度， 摇匀， 即得 lmg/ml的内标储备液； 再稀释成浓度为 5、 10、 25、 50、 100、 225μ^πά 的葛根素甲醇系列标准溶液， 各取 ΙΟμΙ加入 190μ1血清中， 加入 ΙΟμΙ 4-羟基苯甲醛内标溶液（l OO g/ml) 振摇混匀， 处理方法同 "实施例 2 血清处理与测定"， HPLC检测， 以葛根素的血药浓度 C为横坐标， 葛根素的峰面积比 A为纵坐标得 到标准曲线方程式为 A=0.0014C+0.076 , 在 83ng/ml〜3750ng/ml范围 内线性关系良好。
实施例 4: 方法的专属性考察
取空白血浆 190μ1, 按 "血清处理与测定" 项下方法操作， 得到 空白样品色谱图（图 1); 将一定浓度的葛根素和内标加入空白血浆， 按上述方法操作， 获得空白血浆加药的色谱图 (图 2); 结果表明， 血 清中内源性物质不干扰葛根素的测定。将药物给予小鼠灌胃后所取的 血清按上述方法操作后测得血药色谱图 (图 3) , 出峰位置与空白血清 加入药物所测色谱图的出峰位置基本相同。
实施例 5 : 方法的精密度和回收率试验
取小鼠空白血清 190μ1三份放于三个 ΕΡ管中，分别加入 10、 50、 l OO g/ml葛根素甲醇标准溶液即为低中高 3个浓度并按 "实施例 2 血清处理与测定" 下方法处理测定后， HPLC测定， 所得药品与内标 的峰面积比代入标准曲线方程计算测得量。以测得量与加入量相比计 算方法回收率(％：)、 标准方差 (SD)、 相对标准方差 (RSD) , 测得结果如 下表 1。 表 1 : 葛根素的方法回收率
Figure imgf000007_0001
绝对回收率的计算： 取小鼠空白血清 190μ1三份放于三个 ΕΡ管 中， 分别加入 10、 50、 lOO g/ml葛根素甲醇标准溶液即为高中低三 个浓度，分别取同量的药物标准品和内标物质两份，一份加到血清中， 另一份加到流动相溶剂中， 测得药物和内标峰面积计算比值。 测得结 果如下表 2、 表 3。
表 2: 葛根素的绝对回收率
力口入量 ng/ml 166.6667 833.3333333 1666.666667
22533 107520 223960
22522 109562 226420 检测葛根素峰面积 21882 109436 227422
25447 103248 227223
22382 91916 227371 平均值 22953.2 104336.4 226479.2
SD 1596.605 2947.663199 1590.92811
RSD 0.069559 0.028251533 0.00702461 对照葛根素面积 28489 121236 312567 葛根素绝对回收率 0.805686 0.860605761 0.72457809 表 3: 内标的绝对回收率
Figure imgf000008_0001
精密度包括日内精密度与日间精密度， 日内精密度是同一天将样 品测定三次， 日间精密度为不同的时间将样品测定三次。 取小鼠空白 血清 190μ1三份放于三个 EP管中， 分别加入葛根素甲醇溶液得到低 中高 3个浓度并按 "实施例 2血清处理与测定" 下方法处理测定， 同 一天将低中高样品测定三次所得结果求的日内精密度，将处理好的样 品三日内每天测定三次， 所得结果为日间精密度， 结果如下表 4、 表 5。
表 4: 葛根素的日内精密度
加入量 (ng/ml)低 600 600 600 平均值 检测血药浓度 （ng/ml) 539.8946 565.2886 576.2302 560.471 1
SD 18.641
RSD 0.03325892
加入量 (ng/ml)中 833.333 833.333 833.333
检测血药浓度 （ng/ml) 794.2862 843.8238 835.3583 824.4894
SD 26.49705
RSD 0.0321375
加入量 (ng/ml)高 2000 2000 2000
检测血药浓度 （ng/ml) 1785.693 1779.954 1748.007 1771.218
SD 20.30482
RSD 0.01 146376 表 5: 葛根素的日间精密度
Figure imgf000009_0001
实施例 6: 稳定性考察
室温放置稳定性考察：取小鼠空白血清 190μ1放于三个 EP管中， 分别加入低、 中、 高葛根素甲醇标准溶液即为高中低（166.6667、 833.3333、 1666.667ng/ml)三个浓度, 分别在低中高三个浓度按 "实 施例 2" 处理血药， 室温放置， 在 0、 3、 9、 15h分别进行检测所得 结果如下表 6。
表 6: 室温放置稳定性
Figure imgf000009_0002
长期低温稳定性考察：取小鼠空白血清 190μ1放于三个 EP管中， 分别加入低、 中、 高葛根素甲醇标准溶液即为高中低 600、 1500、 2400ng/ml)三个浓度， 分别在低中高三个浓度按 "实施例 2" 处理血 药， -20°C保存， 并在第 1、 2、 3天分别进行检测所得结果如下表 7。 表 7: 长期低温稳定性
Figure imgf000010_0001
反复冻融稳定性考察：取小鼠空白血清 190μ1放于三个 EP管中 分别加入低、 中、 高葛根素甲醇标准溶液即为高中低 (600、 1500、 2400ng/ml)三个浓度， 分别在低中高三个浓度按 "实施例 2" 处理血 药， 放于 -80°C反复冻融两次， 进行检测所得结果如下表 8。
表 8: 反复冻融稳定性
Figure imgf000010_0002
实施例 7: 动物实验
以水与葛根素混合灌胃为对照组， 处理组分别为新鲜蚕血组、模 拟脂质体组、 冻干蚕蛹粉组、 蚕蛹油组， 其中对照组、 新鲜蚕血组、 蚕蛹油组浓度均为 10mg/ml, 即 10mg葛根素（网化化学 （济南）有限 公司）溶于 lml溶剂 ((水、新鲜蚕血 (取 50g蚕蛹研磨捣碎， 500rpm离心 1 Omin后取上清即得)、 蚕蛹油（国家粮食储备局西安油脂科学研究设 计院))中； 模拟脂质体组中， 由新鲜蚕血、 无水乙醇、 羧甲基纤维素 钠及水 (新鲜蚕血： 无水乙醇： 0.5%羧甲基纤维素钠溶液： 水的体积 比为 2: 1 : 4: 0.5)混合成溶液， 然后葛根素以 10mg/ml浓度溶于此 溶液中； 冻干蚕蛹粉组中， 葛根素与蚕蛹粉 (浙江中奇生物药业股份 有限公司）以 l :5(m/m)混合后按葛根素以 10mg/ml溶于 0.5%羧甲基纤 维素钠溶液中。 小鼠灌胃量为 0.2ml/10g (每次)， 各个摸索组分灌胃 后 30min取血， 检测结果见图 4, 寻找到最佳组分蚕蛹油， 以蚕蛹油 为辅料混合葛根素后灌胃并在 5min、 10min、 20min、 3 Omin, 40min、 lh、 1.5h、 2h、 2.5h、 3h、 4h、 6h、 8h、 1 Oh时间点均六只小鼠取血。 血样处理方法同标准曲线， 高效液相色谱测定， 测的面积带入标准曲 线求的浓度。
实施例 8: 用 3p89软件计算各个参数及生物利用度倍数 所检测的血药浓度见图 5, 数据如下表 9。
表 9:
Figure imgf000011_0001
代入 3p89软件 (实用药动学计算程序是中国药理学会数学专业委 员会受国家卫生部药品审评办公室的委托集体编制的，在国内应用很 广泛， 可处理各种用药途径的线性和非线性药动学模型， 给出有关的 药动学参数及各种图表的详细结果)后计算结果如下表 10。
表 10:
Figure imgf000011_0002
其中， Tmax: 药峰时间； Cmax: 药峰浓度； AUC: 血药浓 度曲线下面积； Ke:表观一级吸收速率常数； T1/2: 半衰期；
AUC DR
相对生物利用度 F 100%
AUCR DT
式中 AUCT和 AUCR分别为服用受试 (对照组)的血药曲线下面积， DT 和 DR分别为受试制剂和参比制 剂的剂量 (此处剂量相同均为 10mg/ml, 0.2ml/10g)。
故 F (相对生物利用度) =4.7275倍。
以上所述， 仅为本发明的优选实施例， 应当指出， 对于本技术中 的普通技术人员来说， 在不脱离本发明的核心技术特征的前提下， 还 可以做出若干改进和润饰，这些润饰和改进也应属于本发明的专利保 护范围。

Claims

权利要求书
1.蚕蛹油在提高小分子药物生物利用度中的用途。
2.蚕蛹油作为药用辅料在提高小分子药物生物利用度中的用途。
3.如权利要求 1或 2所述的用途， 其特征在于， 所述小分子药物 以与蚕蛹油混合的形式给予受试者。
4.如权利要求 3所述的用途， 其特征在于， 所述小分子药物与蚕 蛹油以 10mg/ml的比例混合， 并研磨 lOmin, 超声 30min, 充分混合 而得到混合物。
5.如权利要求 4所述的用途， 其特征在于， 所述混合物给予受试 者的剂量以药物与蚕蛹油的混合物计为 0.2ml/l Og体重。
6.如权利要求 5所述的用途， 其特征在于， 所述混合物通过灌胃 的方式给予受试者。
7.如权利要求 6所述的用途， 其特征在于， 所述受试者为小鼠。
8.如权利要求 1-7 任一项所述的用途，其特征在于，所述小分子
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