芒柄花素磺酸钠在脑缺血再灌注大鼠脑组织的药动学研究

目的建立脑组织样品中芒柄花素磺酸钠的高效液相色谱分析方法,确定芒柄花素磺酸钠在脑缺血再灌注大鼠脑内的量–时关系,明确芒柄花素磺酸钠在模型大鼠脑组织中的吸收分布情况。方法制备大鼠脑缺血再灌注模型,随机分为10组,每组各5只。于脑缺血1 h及再灌注3.5 h ip芒柄花素磺酸钠注射液20 mg/kg,并于给药后15、30、45、60、90、120、150、180、240、300 min实施心脏灌流后取脑组织匀浆。样品处理后,采用HPLC法检测各时间点脑组织中芒柄花素磺酸钠的质量浓度。采用药动学软件Win Non Lin 6.3对数据进行分析,计算药动学参数。结果模型大鼠缺血1 h再灌注4 h时给药,脑组织中芒柄花素磺酸钠含量最高。芒柄花素磺酸钠在模型大鼠脑组织内药动学参数t1/2、Cmax、AUC0-t分别为(261.18±49.33)min、(1.56±0.37)μg/m L、(93.60±7.13)min·μg/m L。结论芒柄花素磺酸钠可以进入脑缺血再灌注大鼠脑组织,滞留时间长。     

芒柄花素磺酸钠在脑缺血再灌注大鼠体内的药动学研究

目的建立芒柄花素磺酸钠血浆样品的高效液相色谱(HPLC)分析方法,并应用该方法对正常大鼠和模型大鼠体内的芒柄花素磺酸钠进行药动学分析;明确芒柄花素磺酸钠在正常、模型大鼠血浆中的吸收分布情况。方法采用Venusil MP C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相乙腈–水(30∶70,水相中加0.1%甲酸),柱温30℃,进样量20.0μL,体积流量1.0 mL/min,检测波长250 nm。制备大鼠脑缺血再灌注模型,将SD大鼠随机分为10组,每组5只大鼠。于大鼠脑缺血1 h及再灌注3.5 h按20 mg/kg ip芒柄花素磺酸钠注射液,并于给药后15、30、45、60、90、120、150、180、240、300 min腹主动脉取血。正常组大鼠分别于给药后5、10、15、30、45、60、90、120、150、180、240、300、480、720min目内眦取血。样品经处理后,采用HPLC法检测各时间点血浆中的芒柄花素磺酸钠浓度。采用药动学软件WinNonLin 6.3进行数据分析,计算药动学参数。结果芒柄花素磺酸钠在0.25～100.0μg/mL线性关系良好。定量下限RSD值为12.15%。日内精密度RSD值均5.0%,日间精密度RSD值均10.0%。提取回收率均80%,RSD值均5.0%。芒柄花素磺酸钠在正常大鼠、模型大鼠血浆内药动学参数为:t_(1/2)分别为(173.13±13.12)min、(117.86±45.05)min,C_(max)分别为(67.23±4.04)μg/mL、(75.22±10.57)μg/mL,AUC_(0-t)分别为(2 493.69±216.98)min·μg/mL、(6 094.52±250.81)min·μg/mL,Vd分别为(1 981.97±134.12)mL/kg、(484.56±167.93)mL/kg,Cl分别为(7.96±0.73)mL·min/kg、(2.87±0.14)mL·min/kg,MRT_(0-t)分别为(76.92±1.54)min、(89.06±0.77)min。结论应用HPLC法测定芒柄花素磺酸钠的血药浓度定量准确、灵敏,芒柄花素磺酸钠在模型大鼠血浆中吸收的量多,单位时间内消除的量少,滞留时间长。     

HPLC法测定黄芪不同炮制品中芒柄花素

目的 建立黄芪药材中黄酮类成分芒柄花素的HPLC测定方法,并考察黄芪不同炮制品中芒柄花素的变化.方法 采用HPLC法测定芒柄花素.色谱柱:Thermo ODS-2 Hypersil (250 mm×4.6 mm,5μm);流动相:乙腈-0.05％冰乙酸水溶液(35∶65):体积流量:1 mL/min;波长:248 nm;柱温:30℃.将黄芪药材分别制成生黄芪、炒黄芪、蜜炙黄芪、盐制黄芪以及酒制黄芪,并比较黄芪不同炮制品中芒柄花素.结果 黄芪不同炮制品中芒柄花素有所差异,其中蜜炙后芒柄花素的量降低幅度较大.结论 该方法简便、准确,适用于黄芪不同炮制品中芒柄花素的定量分析.     

4个黄芪异黄酮类化合物对PC12细胞分化的影响

目的探究毛蕊异黄酮、刺芒柄花素、毛蕊异黄酮苷、芒柄花苷对PC 12细胞分化的影响。方法培养PC 12细胞,与不同浓度的神经生长因子(NGF)、毛蕊异黄酮、刺芒柄花素、毛蕊异黄酮苷、芒柄花苷共处理5 d,1次/d,连续3次,观察PC 12细胞突起向外生长的情况。同时,用免疫荧光染色检测上述化合物对β微管蛋白(βⅢ-tubulin)表达的影响。结果与对照组相比,毛蕊异黄酮、刺芒柄花素、毛蕊异黄酮苷和芒柄花苷(0.01~10.00μmol/L)未能促进PC 12细胞突起向外生长和β微管蛋白的表达。结论毛蕊异黄酮、刺芒柄花素、毛蕊异黄酮苷、芒柄花苷不能促进PC 12细胞分化。     

芒柄花素药理作用的研究进展

芒柄花素属于异黄酮类化合物,是黄芪中异黄酮类化合物的代表成分,具有抗炎、抗氧化、抗肿瘤、雌激素样作用.对芒柄花素保护脑血管系统、保护心血管系统、抗肿瘤作用、对糖尿病心肌病的保护作用等进行了总结,以期为芒柄花素的基础研究和临床应用提供依据.     

蒙古黄芪中芒柄花素含量的测定及黄酮类成分提取工艺的优化

目的建立芒柄花素的含量测定方法,优选黄芪中黄酮类成分的提取工艺。方法采用高效液相色谱-紫外检测法,建立芒柄花素的含量测定方法,以芒柄花素含量为指标,用正交设计法对影响超声提取的因素如乙醇浓度、乙醇用量、提取次数和超声时间进行考察,优化提取工艺。结果芒柄花素在1～40μg/ml范围内线性关系良好,以峰面积(A)对浓度(C)进行线性回归,方程为A=212096C+3958.7,r2=0.9999。日内、日间相对标准差(RSD)均<5%;平均回收率均在98.0%～102.0%;优化提取条件为加15倍量75%乙醇,连续提取3次,每次20min。结论所建立的含量测定方法可用于蒙古黄芪中芒柄花素的含量测定,优化的提取工艺具有效率高和操作简便等优点。     

HPLC法同时测定黄芪中4种黄酮类成分的含量

目的:建立以高效液相色谱法同时测定11个产地黄芪中毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮和芒柄花素4种黄酮类成分含量的方法,从有效成分含量探讨药材道地性内在因素。方法:色谱柱为Zorbax Eclipse XDB-C1(8250mm×4.6mm,5μm),流动相为乙腈-0.2%甲酸水溶液(梯度洗脱),检测波长为280nm。结果:毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮和芒柄花素的检测浓度分别在0.0051～0.510、0.0050～0.300、0.0049～0.294、0.0046～0.276mg·mL-1范围内与各自峰面积积分值呈良好的线性关系(r均为0.9999)。内蒙古、山西等4个产地的黄芪中毛蕊异黄酮苷和芒柄花苷的含量较高;河北安国、赤城和甘肃定西产黄芪中毛蕊异黄酮和芒柄花素含量较高。结论:本方法简便、快速、准确,试验结果与黄芪本草考证、道地沿革的文献基本相符。     

HPLC波长切换法同时测定参芪膏中6个主要成分的含量

目的建立HPLC波长切换法同时测定参芪膏中党参炔苷、丁香苷、毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮和芒柄花素的含量。方法采用Zorbax XDB-C18色谱柱(4.6 mm×250 mm,5.0μm),以乙腈-甲醇(9∶1)(A)-0.1%甲酸溶液(B)为流动相,进行梯度洗脱,流速0.9 mL/min,柱温30℃,进样量为10μL,检测波长:266 nm(党参炔苷、丁香苷)、254 nm(毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、芒柄花素)。结果党参炔苷、丁香苷、毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮和芒柄花素6个成分分别在15.49~309.80、2.15~43.00、5.66~113.20、4.89~97.80、3.28~65.60、12.06~241.20μg/mL范围内峰面积与浓度呈良好的线性关系,相关系数分别为0.999 7、0.999 2、0.999 5、0.999 3、0.999 6、0.999 9;平均加样回收率及相应的RSD分别为98.07%(0.61%)、99.09%(1.53%)、99.44%(1.33%)、97.86%(1.28%)、99.95%(1.04%)、97.19%(0.58%)。结论所建立的方法快速,灵敏度高,准确度高,专属性好,为参芪膏的质量控制提供依据。     

RP-HPLC法测定鸡血藤中芒柄花素的含量

目的建立测定鸡血藤药材中芒柄花素的RP-HPLC法。方法采用Kromasil C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(体积比为35∶65),流速为1 mL.min-1,检测波长为254 nm,柱温为30℃。结果芒柄花素在0.5~15.4 mg.L-1内线性关系良好(r=0.999 8,n=6),平均回收率为98.6%(RSD=1.6%,n=9)。结论RP-HPLC法可用于鸡血藤中芒柄花素的含量测定。     

芒柄花素衍生物的合成及其初步生物活性

设计合成了８个芒柄花素（７－羟基－４’－甲氧基异黄酮）衍生物,其中７个为新化合物,并进行了体外初步抗骨质疏松活性实验,结果显示这些化合物对体外培养的兔骨髓成骨细胞的生长有较好的促进作用。     

硝硫氰胺水溶性衍生物的合成

目的 将脂溶性抗血吸虫病药物硝硫氟胺转化为水溶性衍生物，简化血吸虫患畜治疗的给药方式。方法 通过化学手段引入水溶性基团进行结构改造。结果 合成数全水溶性衍生物。结论 硝硫氰胺的氨基酸衍生物的钠盐水溶性良好。     

Design,synthesis, and biological evaluation of target water‐soluble hydroxamic acid‐based HDACi derivatives as prodrugs

Four compounds T1 , T2 , T3 , and T4 were designed and synthesized as Vorinostat and Belinostat derivatives being the target water‐soluble prodrugs. The water solubility of Vorinostat derivatives, T1 and T2 , exhibited 400‐ to 600‐fold higher than that of Vorinostat, and Belinostat derivatives, T3 and T4 , showed 600‐ to 750‐fold higher than that of Belinostat. Four compounds were evaluated for their inhibitory activities against tumor cell lines HT‐29 and Hut‐78 in the absence or presence of β‐D‐glucuronidase. The inhibitory effects of T1 and T2 were comparable to Vorinostat in the presence of β‐D‐glucuronidase, but were higher than 10 μM in the absence of β‐D‐glucuronidase. Therefore, T1 and T2 are promising candidates for in vivo investigations with high potential to be the target water‐soluble prodrugs. IC₅₀ values of Belinostat derivatives T3 and T4 were not affected by β‐D‐glucuronidase, but T3 and T4 had the excellent cell proliferation inhibition on Hut‐78.     

Increasing antitumor activity in vivo by enhancing acridine dimer solubility with salt preparations

The potent activities of many anticancer agents have been demonstrated by in vitro assays. However, their poor solubility may result in diminishing anticancer activities in vivo. Previously, we synthesized a series of bisacridine derivatives shown to be potent in cytotoxicity and DNA intercalating activity in vitro. Initially, the compound 1, (N-(6-chloro-2-methoxy-acridin-9-yl)-N′-[3-(6-chloro-2-methoxy-acridin-9-ylamino)-propyl]-propane-1,3-diamine), is insoluble in polar solvent and does not reveal antihuman COLO 205 solid-tumor activity in vivo. To enhance its solubility, three salt forms (CH₃COOH, CH₃SO₃H, and CF₃COOH) of compound 1 were synthesized and their solubility was found to be greatly improved compared with that of the free base. Among these salts, the compound 1 · (tris)acetate salt has shown good solubility in H₂O and 2.5% Cremophor (v/v) and demonstrated anti-COLO205 solid-tumor activity in vivo.     

Hypolipidemic activity of <Emphasis Type="Italic">Symplocos cochinchinensis</Emphasis> S. Moore leaves in hyperlipidemic rats

The hypolipidemic activity of Symplocos cochinchinensis S. Moore leaves was studied in Triton WR-1339- and high fat diet-induced hyperlipidemic rats. In Triton WR-1339-induced hyperlipidemic rats, the hexane extract (250 and 500 mg/kg) exerted a significant (P < 0.01) lipid-lowering effect compared to ethyl acetate and methanol extracts, as assessed by the reversal of the plasma levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). In high fat diet-fed hyperlipidemic rats, the hexane extract (250 and 500 mg/kg) caused the lowering of lipid levels in the plasma and liver. The hypolipidemic activity of S. cochinchinensis leaves was compared with fenofibrate, a known lipid-lowering drug, in both models.     

A High Throughput Protein Formulation Platform: Case Study of Salmon Calcitonin

Purpose  The feasibility of using high throughput spectroscopy for characterization and selection of physically stable protein formulations was studied. Materials and Methods  A hundred aqueous formulations of salmon calcitonin (sCT) were prepared using 20 buffer compositions. The solutions had pH values between 2.5 and 10.5. The stability of the sCT formulations was analyzed over 1 week by the following assays: (1) protein concentration, (2) volume control by measuring pathlength, (3) turbidity (absorbance at 350 nm), (4) intrinsic tyrosine fluorescence, (5) 1-anilino-naphthalene-8-sulfonate (ANS) fluorescence, (6) Nile Red fluorescence. Addition of the dyes (Nile Red and ANS) was used to study protein conformational changes. Results  After 1 day, 27 out of the 100 formulations of salmon calcitonin were stable. After 7 days, 12 stable sCT formulations remained. The best salmon calcitonin formulation was in 10 mM sodium acetate buffer with pH values between 3.5 and 5.5. Conclusions  The findings are in accordance with the sCT formulations that were patented and used commercially. This can be considered as a proof of concept for the high throughput protein formulation platform. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Superporous hydrogels based on blends of chitosan and polyvinyl alcohol as a carrier for enhanced gastric delivery of resveratrol

Resveratrol exhibits a number of pharmacological properties, notably antioxidant, anti-inflammatory and anti-cancer activities which are beneficial for the treatment of gastric diseases. However, the poor aqueous solubility and rapid metabolism are the important limitations in clinical uses. Superporous hydrogels (SPHs) based on chitosan/PVA blends were developed as a carrier for resveratrol solid dispersion (Res_SD) to increase the solubility and achieve sustained drug release in the stomach. The SPHs were prepared by gas forming method using glyoxal and sodium bicarbonate as cross-linking agent and gas generator, respectively. The solid dispersions of resveratrol with PVP-K30 were prepared by solvent evaporation and incorporated into the superporous hydrogels. All formulations showed rapid absorption of simulated gastric fluid and reached the equilibrium swollen state within a few minutes. The water absorption ratio and mechanical strength of SPHs were predominantly affected by the chitosan content, with maximum values at 1400 % and 375 g/cm², respectively.The Res_SD-loaded SPHs exhibited good floating properties and SEM micrographs revealed a highly interconnected pores structure with size around 150 μm. Resveratrol was efficiently entrapped within the SPHs at levels between 64 and 90 % w/w and efficient drug release was sustained over 12 h dependent on the concentration of chitosan and PVA. The Res_SD-loaded SPHs exhibited slightly less cytotoxic efffect towards AGS cells than pure resveratrol. Furthermore, the formulation showed similar anti-inflammatory activity against RAW 264.7 cells compared with indomethacin.     

3-氧代齐墩果酸氨基酸偶联物的合成、水溶性的测定及抗肿瘤活性研究

目的设计合成3-氧代齐墩果酸氨基酸偶联物，并测试其水溶性和体外抗肿瘤活性。方法利用HPLC法测定具有代表性结构的偶联物的水溶性，并采用MTT法测试这些偶联物对KB和KB／V两种癌细胞的抑制活性。结果共合成9个新化合物，其结构经核磁共振氢谱和质谱确证。结论被测试的6个偶联物的水溶性与3-氧代齐墩果酸相比有不同程度的改善，水溶性大小顺序是5e〉5g〉5c〉5d〉5a〉5b和3-氧代齐墩果酸。初步体外抗肿瘤活性结果表明，水溶性较高的偶联物（5e，5g，se）仍保持抗肿瘤活性。     

Kinetic solubility and dissolution velocity of rutin nanocrystals

Lyophilized rutin nanocrystals were intensively evaluated regarding their physicochemical properties with respect to particle size analyses, crystallinity, kinetic solubility and dissolution behavior. The particle size was determined by photon correlation spectroscopy (PCS) and laser diffraction (LD). DSC and X-ray diffraction were used to study the crystalline state of rutin nanocrystals. In a period of 1 week, the kinetic solubility was determined using a shaker at 25 °C. DSC and X-ray diffraction analyses showed that lyophilized rutin nanocrystals prepared by high pressure homogenization remained in crystalline state. Lyophilized rutin nanocrystals could be re-dispersed completely in water and the kinetic solubility in water increased to 133 μg/ml.. Lyophilized rutin nanocrystals were almost completely dissolved within 15 min in water, buffer of pH 1.2 and buffer of pH 6.8. In contrast, only 70% of rutin raw material (rutin microcrystals) was dissolved within 15 min. The superior physicochemical properties of rutin nanocrystals should overcome the absorption problem in the gastrointestinal tract and increase the bioavailability.     

Effect of high fluoride and high fat on serum lipid levels and oxidative stress in rabbits

The purpose of this study was to explore the effects of high fluoride and high fat on triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), total antioxidant capacity (T-AOC), lipid peroxide (LPO) and malondialdehyde (MDA) in rabbits. A factorial experimental design was used, with two factors (fluoride and fat) and three levels. Seventy-two male rabbits were randomly assigned into nine groups according to initial weight and serum lipid levels. The rabbits were fed with basic feed, moderate fat feed or high fat feed and drank tap water, fluoridated water at levels of 50 and 100 mg fluorion/L freely. Biological materials were collected after 5 months, and serum lipid, T-AOC, LPO, and MDA levels were then measured. Using these data, the separate and interactive effects of high fluoride and high fat were analyzed. High fluoride and high fat both increased serum levels of TC, HDL-C and LDL-C significantly (P < 0.05), and there was also a synergistic effect between high fluoride and high fat (P < 0.05). High fluoride and high fat had different effects on TG levels: high fat significantly increased TG levels (P < 0.01) whereas high fluoride had nothing to do with TG levels (P > 0.05). High fat significantly elevated LPO and MDA levels and lowered T-AOC levels in serum (P < 0.05). Similarly, high fluoride significantly increased LPO and MDA levels in serum (P < 0.05). However, there was no interactive effect between high fat and high fluoride on these indexes. In summary, high fluoride and high fat increased serum TC and LDL-C levels individually and synergistically, and this would cause and aggravate hypercholesterolemia in rabbits. At the same time, high fluoride and high fat both made the accumulation of product of oxidative stress in experimental animals.     

Design,Synthesis, and Biological Evaluation of Scutellarein Carbamate Derivatives as Potential Multifunctional Agents for the Treatment of Alzheimer's Disease

A series of scutellarein carbamate derivatives were designed and synthesized based on the multitarget‐directed drug design strategy for treatment of Alzheimer's disease. Their acetylcholinesterase and butyrylcholinesterase inhibitory activities, antioxidant activities, metals chelation, and neuroprotective effects against hydrogen peroxide‐induced PC12 cell injury were evaluated in vitro. The preliminary results indicated that compound 7b exhibited good inhibitory potency toward AChE and BuChE with IC₅₀ values of 1.2 ± 0.03 μm and 22.1 ± 0.15 μm , respectively, possessed the strong antioxidant potency (10.3 trolox equivalents), as well as acted as a selective metal chelator and neuroprotective agent. Furthermore, 7b could improve memory impairment induced by scopolamine, ethanol, and sodium nitrite using the step‐down passive avoidance task in vivo and could remarkably decrease the activity of acetylcholinesterase in mice brain. This study indicated that 7b could be considered as a potential multitarget agent against AD.