喜树碱-甾体新缀合物的设计合成及抗肿瘤活性

目的:设计合成喜树碱-甾体新缀合物,考察其抗肿瘤活性。方法:以喜树碱或者7-羟甲基喜树碱和一系列甾体酸为原料,采用EDCI-DMAP偶合法,合成喜树碱-甾体新缀合物;以喜树碱为阳性对照,采用MTT法进行初步的体外抗HepG2细胞活性筛选。结果:喜树碱-甾体新缀合物的结构通过1H-NMR,质谱和元素分析确认;化合物2a,3d对HepG2细胞的抑制活性明显高于喜树碱。结论:合成的喜树碱-甾体缀合物2a,3d有较好活性,值得进一步研究。     

11β，17α-二羟基-17-丙酰基-1，4-雄甾二烯-3-酮的合成

目的 制备17-丙酰基功能化的甾体化合物11β,17α-二羟基-17-丙酰基-1,4-雄甾二烯-3-酮.方法 以泼尼松龙为原料,经5步反应合成目标物.结果 和结论 总收率为67.4%,通过NMR、MS等确证了中间体和目标物的结构.     

氧甲氢龙衍生物的合成

目的：设计合成氧甲氢龙的衍生物。方法：以3β-羟基-5α-雄甾烷-17-酮为原料经氧化开环、还原、缩合得到中间体17β-羟基-A-失碳-5α-雄甾烷-2-酮,此中间体与不同的酰氯反应生成的一系列C17位酰化产物再经过Baeyer -Villiger反应得到氧甲氢龙衍生物系列A;以3β-羟基-5α-雄甾烷-17-酮为原料与不同酰氯反应生成氧甲氢龙衍生物系列B;以3β-羟基-5α-雄甾烷-17-酮的酰化产物为原料经过Baeyer-Villiger反应得到氧甲氢龙衍生物系列C。结果与讨论 ：合成了3个系列22个未见报道的新化合物,目标化合物的结构均经H1-NMR、IR、MS谱确证。     

部分A-失碳-5α-雄甾烷类化合物的合成

合成A-失碳-5α-雄甾烷-2,17-双酮肟(Ⅳa),A-失碳-5α-雄甾烷-2,17-双酮苯腙(Ⅳc),A-失碳-5α-雄甾烷-2,17-双酮-2-腙(Ⅳb)以及2ξ-乙炔基-A-失碳-5α-雄甾烷-2ξ,17β-二醇双丙酸酯(Ⅷb)及其17-肉桂酸酯(Ⅷa)。用元素分析、红外光谱证实了这些化合物的结构,其中化合物Ⅳb和Ⅷa尚未见文献报道。这些化合物正在作小白鼠的抗生育活性试验。     

17-肟基雄甾烷类化合物的合成及抗肿瘤活性研究

目的设计合成一系列17-肟基雄甾烷类化合物,并进行体外抗肿瘤活性测试。方法以去氢表雄酮为起始原料,通过PCC氧化、选择性还原以及选择性肟化,合成一系列17-肟基雄甾烷类化合物;采用MTT法对合成的化合物进行体外抑制肿瘤细胞增殖活性测试。结果与结论设计合成了5个17-肟基取代雄甾烷类化合物,活性研究表明,这5个化合物对人鼻咽癌CNE细胞株和人胃癌SGC-7901细胞株表现出弱的抑制活性。     

PPARα/γ双重激动剂TZD18与脂质代谢的研究进展

噻唑烷二酮（TZD）包括一系列具有2,4-噻唑烷二酮结构的化合物.它们均具有不同的侧链取代基,因而药理特点各有不同.最近发现一种化合物TZD18,一种兼具有改善胰岛素抵抗和降低甘油三酯、胆固醇作用的PPARα/γ的双重激动剂.本文综述了TZD18在脂质代谢方面的研究进展.     

1-（1，4-苯并二噁烷-2-羰基）-4-取代芳氧烷基哌嗪类α1受体拮抗剂的合成及其生物活性

目的寻找新的1,4-苯并二烷类α1受体拮抗剂。方法选用1,4-苯并二烷为母核,以儿茶酚和取代苯酚为原料,分别经关环、水解、成酰胺、取代、成盐等反应合成目标化合物,并测定该类化合物α1受体拮抗活性。结果与结论设计合成了13个新化合物,结构经ESI-MS、1H-NMR、IR及HR-MS确证。初步药理活性实验表明,其中8个目标化合物的pA2值大于6.00,具有良好的α1受体拮抗活性,有进一步研究的价值。     

2-芳亚胺基-4-噻唑烷酮类化合物的合成及生物活性

目的设计、合成新的2-芳亚胺基-4-噻唑烷酮类化合物并研究其对一氧化氮合酶(NOS)的抑制活性。方法运用N-氯乙酰-1,2,3,4-四氢异喹啉或N-氯乙酰邻苯二甲酰亚胺与取代硫脲反应，简便地合成了15个新的2-芳亚胺基-4-噻唑烷酮类化合物，测试新合成的目标化合物的NOS抑制活性。结果和结论合成了15个新的2-芳亚胺基-4-噻唑烷酮类化合物，大部分反应收率大于65%。所有化合物的结构用¹H NMR，IR，MS及元素分析进行了表征。初步药理筛选结果表明，部分化合物具有NOS抑制活性。     

1-(1,4-苯并二(口恶)烷-2-羰基)-4-取代芳氧烷基哌嗪类α1受体拮抗剂的合成及其生物活性

目的 寻找新的1,4-苯并二(口恶)烷类α1受体拮抗剂.方法 选用1,4-苯并二(口恶)烷为母核,以儿茶酚和取代苯酚为原料,分别经关环、水解、成酰胺、取代、成盐等反应合成目标化合物,并测定该类化合物α1受体拮抗活性.结果与结论 设计合成了13个新化合物,结构经ESI-MS、1H-NMR、IR及HR-MS确证.初步药理活性实验表明,其中8个目标化合物的pA2值大于6.00,具有良好的α1受体拮抗活性,有进一步研究的价值.     

2-氨基噻唑类化合物的合成

目的合成2-氨基噻唑类化合物。方法溴化铜在乙酸乙酯/氯仿中分别溴化芳基烷基酮及脂肪酮,得到的α-溴化产物,不经分离直接与硫脲环合制得相应的2-氨基噻唑类化合物。结果合成收率为87.4%~98.9%。目标物2-氨基噻唑类化合物的结构经红外光谱、核磁共振谱和质谱确证。结论该合成路线可行,收率高,得到的10个化合物中有4个是未见报道的新化合物。     

The proteomic study and the target discovery of W026B,a new compound with brain protective effect

W026B is a new compound that has a protective effect on cerebral ischemia reperfusion (I-R) injury in mice, while its specific mechanism is still unknown. In this study, proteomics was used to observe the effect of W026B on protein expression in brain I-R tissue, and to reveal its potential target. A total of 42 significantly altered proteins were identified in both brain I-R model and W026B treatment from 4852 proteins detected by proteomics, and most of these proteins were related to immunity and inflammation, metabolism, neuroprotection as well as cell proliferation and cell structure. Western blotting analysis showed that three out of five selected proteins showed consistent alteration with the proteomics. Regulator of G protein signaling 17 (RGS17) was selected for further study, and its knockdown by siRNA RGS17 aggravated brain injury and abolished the protective effect of W026B. W026B could bind with RGS17 (K_D: 6.04×10^–6 mol/L). The knockdown of RGS17 aggravated Neuro-2a cell damage induced by group I metabotropic glutamate receptors (mGluRs) agonist, and abolished the protective effect of W026B. In conclusion, W026B protected brain against I-R injury by affecting diverse proteins. RGS17 might be one of its targets and a potential therapeutic target of brain I-R injury. The upstream receptor of G protein, which was regulated by RGS17 and affected by W026B, might be group I mGluRs. This study provided useful evidence for the further R&D and the potential clinical application of W026B.     

盐酸麻黄素植入膜剂的研制与临床应用

目的 :研制盐酸麻黄素植入膜剂 ,为临床治疗慢性、顽固性支气管哮喘开发新剂型与给药途径。方法 :研究盐酸麻黄素植入膜剂的处方、制备、质量控制 ,并通过体外释药试验评价缓释膜作用及动物试验考查其安全性。结果 :盐酸麻黄素植入膜剂的含药膜基质以明胶为佳 ,延缓释药膜基质以PVA17 -99 为佳。含量测定用紫外分光光度法。体外释药试验表明 ,本品缓释效果好 ,植入穴位无任何不良反应。临床应用疗效可靠、作用持久、安全性好。结论 :盐酸麻黄素植入膜剂植入双定喘穴具穴位疗法和药物靶区定向缓释的双重治疗作用 ,是一个有前途的新制剂     

孤儿核受体RORγ拮抗剂的合成及其初步活性研究

目的设计合成新型维甲酸受体相关孤儿受体γ(RORγ)小分子拮抗剂,并检测其在细胞水平上对蛋白活性的影响。方法 2-氟苯胺与六氟丙酮发生取代反应,再经重氮化反应,叠氮化钠亲核进攻生成叠氮化合物,最终与炔基官能团发生分子间环加成反应生成三唑类氮杂环,共合成两类柔性不同的连接基团组成的目标化合物。采用荧光素酶报告基因实验(luciferase reporter gene assay)测定目标化合物对RORγ细胞水平的抑制活性。结果与结论合成了15个未见文献报道的新化合物,其结构经1H-NMR和ESI-MS谱确证。活性评价结果显示,多个化合物在细胞水平上对RORγ有抑制活性,其中化合物8a表现出最强的抑制活性,其抑制活性略优于阳性对照化合物,由此推论此类化合物可能具有潜在的抗炎作用。     

AlphaLISA方法测定抗白介素-17受体单抗生物学活性

目的：利用AlphaLISA方法建立抗白介素-17受体单抗的生物学活性测定方法。方法：以人包皮成纤维细胞系作为靶细胞,通过AlphaLISA检测系统建立抗IL-17R单抗的生物学活性测定方法,根据四参数拟合分析计算抗IL-17R单抗的相对效价,并对该方法进行了验证。结果：抗IL-17R单抗在该方法中存在量效关系,并且符合四参数方程：y=（A-D）/[1+（x/C）^B]+D。6批抗IL-17R单抗经3次测定,相对效价平均值在（84.93±3.12）%~（107.51±1.66）%,相对标准偏差小于5%。2批回收率样品经3次测定,回收率分别为（100.66±13.65）%和（115.69±3.85）%。结论：本研究利用AlphaLISA方法在国内首次成功建立重复性好、准确性高的抗IL-17R单抗的生物学活性测定方法。     

黄芩中有机氯类农药残留检测方法研究

目的建立黄芩中农药多残留量的新检测方法,考察陕西不同地区黄芩药材中12种农药残留情况。方法采用DB-17MS毛细管柱(30m×0.25mm×0.25μm),用气相色谱-质谱串联技术(GC-MS)进行目标成分的定量测定。结果建立了气相色谱-质谱串联法测定黄芩中农药多残留的方法,通过对陕西省不同地区黄芩药材中农药进行检测,发现尚无超标的农药。结论该方法严谨可靠,操作简便,能较准确地反映测定结果,与国家现有标准的方法相比,实验时间短,灵敏度高,减少了人为因素的影响。     

A new class of nonsteroidal aromatase inhibitors: design and synthesis of chromone and xanthone derivatives and inhibition of the P450 enzymes aromatase and 17 alpha-hydroxylase/C17,20-lyase

Aromatase (P450arom) is a target of pharmacological interest for the treatment of breast cancer. In this paper, we report the design, synthesis, and in vitro biological evaluation of a series of new (di)benzopyranone-based inhibitors of this enzyme. The design of the new compounds was guided by a CoMFA model previously developed for a series of nonsteroidal aromatase inhibitors. Both the chromone and the xanthone nuclei were taken as molecular skeletons, and the functions supposed to be critical for binding to the aromatase active site - a heterocyclic ring (imidazole or 1,3,4-triazole) linked to the aromatic moiety by a methylene unit and an H-bond accepting function (CN, NO(2), Br) located on the aromatic ring at a suitable distance from the heterocyclic nitrogen carrying the lone pair--were attached to them. The chromone, xanthone, and flavone derivatives were prepared by conventional synthetic methods from the appropriate methyl analogues. Aromatase inhibitory activities were determined by the method of Thompson and Siiteri, using human placental microsomes and [1 beta,2 beta-(3)H]testosterone as the labeled substrate. All the compounds were also tested on 17 alpha-hydroxylase/C17,20-lyase (P450 17), an enzyme of therapeutic interest for the treatment of prostatic diseases. The goal to find new potent inhibitors of aromatase was reached with the xanthone derivatives 22d,e (IC(50) values 43 and 40 nM, respectively), which exceeded the potency of the known reference drug fadrozole and also showed high selectivity with respect to P450 17. Moreover, compounds 22g-i based on the same xanthonic nucleus showed fairly high potency as P450 17 inhibitors (IC(50) values 220, 130, and 42 nM, respectively). Thus, they might be new leads for the development of drug candidates for androgen-dependent diseases.     

Synthesis and biological evaluation of a new class of geldanamycin derivatives as potent inhibitors of Hsp90

The heat shock protein Hsp90 has increasingly become an important therapeutic target especially for treatment of cancers. Inhibition of the ATPase activity of Hsp90 by natural products (e.g., 17-allylaminogeldanamycin or radicicol) leads to the ubiquitination of oncogenic client proteins such as Her-2, Raf-1, and p-Akt followed by their proteasomal degradation. Hsp90 inhibitors simultaneously target multiple oncogenic proteins and provide an advantage for cancer therapy due to the potential for increased efficacy and overcoming drug resistance. In an effort to convert geldanamycin into a druglike compound with better pharmacokinetic properties and efficacy in human tumor xenograft models, geldanamycin was derivatized on the 17-position to prepare new analogues such as 17-geldanamycin amides, carbamates, and ureas and 17-arylgeldanamycins. All the compounds were first evaluated ex vivo using a cell-based Her-2 degradation assay and in vitro using biochemical assays that measure recombinant Hsp90 (rHsp90) competitive binding and changes in rHsp90 conformation. In addition, we confirmed the selectivity of geldanamycin analogues for Hsp90 derived from tumor cells using a novel cell lysate binding assay.     

GLC determination of 17 alpha-ethynylestriol 3-cyclopentyl ether.

A rapid, sensitive, and accurate GLC method of analysis of a new estrogenic drug, 17 alpha-ethynylestriol 3-cyclopentyl ether, was developed. The drug and the internal standard, tetratriacontane, are dissolved in chloroform, and an aliquot is heated with N-trimethylsilylimidazole at 80 degrees for 30 min. The silylated sample is chromatographed using a column packed with 1% methyl vinyl silicone gum on Gas Chrom Q. Quantitation is achieved by computer calculation of the peak area ratios. The observed peak is the 16alpha,17beta-bistrimethylsilyl derivative of the new drug substance. The GLC method was applied to the quantitative determination of the estrogenic compound in a tablet formulation containing 25 mug/tablet.     

Bioinformatics Prediction and In Vitro Analysis Revealed That miR‐17 Targets Cyclin D1 mRNA in Triple Negative Breast Cancer Cells

Breast cancer is one of the most prevalent malignancies among women worldwide. Triple negative breast cancer (TNBC) is a type of breast cancer in which estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER‐2) are not expressed. There is no targeted therapy for this type of cancer, and available therapies have poor therapeutic effects. Performing a preliminary research, we selected cyclin D1 (CCND1) gene of Wnt signaling pathway which is a target of miRNAs, a promising set of biomolecules in diagnosis and treatment of breast cancer. In this study using bioinformatic analyses, miR‐17 was selected as it targets the 3′UTR of CCND1 gene with the highest score. Luciferase assay results also confirmed the bioinformatic prediction. Decreased expression of miR‐17 in MDA‐MB‐231 cell line was observed using qRT‐PCR method. After lentiviral transduction of miR‐17 to the target cells, gene expression analysis showed decreased expression of CCND1 gene. We found miR‐17 as an attractive molecule that after intensive research can probably be used as a biomarker in TNBC.     

一种腺病毒载体疫苗的体外生物活性和细胞毒性研究

目的探讨一个以腺病毒为载体的人用疫苗(代号LMP-Ad)在传代动物细胞株上的表达能力和细胞毒性,为动物试验提供依据。方法受试物LMP-Ad以不同感染复数(MOI=3000、1000、300、100和30)感染非洲绿猴肾细胞(Vero)、幼仓鼠肾细胞(BHK)、小鼠成纤维细胞(L929)和人胚肺二倍体细胞(KMB17)4种传代细胞株后,用免疫细胞化学染色法检测目的蛋白LMP在细胞膜上的表达情况;在MOI=3×105、3×104、3×103、3×102和30时,用MTT法和FCM检测细胞相对增殖率(RGR)和细胞死亡率。结果在4种细胞中均检测到目的蛋白的表达,其中BHK细胞呈弱阳性,而Vero和KMB17在MOI低至30时仍较好地表达目的蛋白;随着感染剂量的提高,4种细胞的RGR下降而死亡率上升,其中以Vero细胞的变化最为明显。结论受试物对上述细胞均具生物活性,并在高感染剂量时表现出对细胞的生长抑制和一定程度的损伤,4种细胞以Vero和KMB17对受试物最为敏感,更适于作为该类新药的体外研究模型。