N-(1-乙氧羰基-3-苯氨甲酰基丙基)甘氨酰甘氨酸衍生物的合成

报道4个N-(1-[1-乙氧羰基-3-(对甲)苯氨甲酰基]丙基甘氨酰｝-N-取代甘氨酸(XI_1～4)和5个1-[1-乙(或甲)氧羰基-3-(对甲)苯氨甲酰基]丙基-4-取代-1,4-哌嗪-2,5-二酮(XII_1～5)共9个估计有血管紧张素转化酶抑制活性化合物的合成和鉴定。所有这些化合物及9个相应的酯(X_1～9)均未见文献报道。药理初试结果,化合物XII₂,XII₅,XI₄和XII₁均有较强降压活性。     

国产醋酸甲地孕酮的质量研究:6-羟基-6-甲基-17α-乙酰氧基黄体酮差向异构体的分离与鉴定

用柱层析及薄层层析对国产醋酸甲地孕酮的杂质进行了分析,共分离出7个杂质点:Ⅰ₁,Ⅰ₂,Ⅰ₃,Ⅰ₄,Ⅰ₅,Ⅰ₆,Ⅰ₇,并对其中Ⅰ₃和Ⅰ₄进行分离纯制及光谱分析,确定其结构为6β-羟基-6α-甲基-17α-乙酰氧基黄体酮及其差向异构体6α-羟基-6β-甲基-17α-乙酰氧基黄体酮。本文报道了薄层层析条件(硅胶GF₂₅₄,展开剂:醋酸乙酯-甲苯=7:3)以及光谱鉴别的依据。     

丁基苯酞抗大鼠大脑皮质神经元氧糖剥夺/复氧损伤及其机制

本研究探讨丁基苯酞抗大鼠大脑皮质神经元氧糖剥夺/复氧损伤及其机制。原代培养大鼠大脑皮质神经元，建立氧糖剥夺/复氧模型(OGD/R)，采用MTT法、酶学检查、免疫组化、RT-PCR等观察丁基苯酞(各浓度组)的保护作用及其机制。在氧糖剥夺4 h/复氧8 h时丁基苯酞各浓度组可增加神经元的细胞活力和减少神经元LDH(乳酸脱氢酶)的释放，可显著降低神经元表达iNOS mRNA(诱生型一氧化氮合酶)和NF-κB(核因子κB) p65蛋白(增加)。不同剂量丁基苯酞(100、 10、 1和0.1 μmol·L^-1)在增加细胞活力、减少LDH释放及降低神经元表达iNOS mRNA等方面，高浓度的作用强于低浓度，且丁基苯酞100 μmol·L^-1组与吡咯烷二硫代氨基甲酸酯(pyrrolidine dithiocarbamate，PDTC) 100 μmol·L^-1组差异显著。在OGD 4 h/R 8 h时丁基苯酞可能抑制iNOSmRNA的表达及NF-κB的活化，从而有效保护氧糖剥夺/复氧中损伤的大脑皮质神经元。     

2-(E)-取代苯亚甲基环戊酮及其Mannich碱盐酸盐类化合物的合成和抗炎、抗癌活性研究

本文设计,合成了2-(E)-取代苯亚甲基环戊酮(Ⅰ)类化合物18个及其Mannich碱盐酸盐(Ⅱ)类化合物11个,其中22个为新化合物。初步药理结果显示:对于角叉菜胶诱发的大鼠足趾水肿,Ⅰ₄,Ⅰ₁₂及Ⅰ₁₃口服有较显著的抑制作用,水溶性Ⅱ类化合物皮下注射有极强的抑制作用,其中Ⅱ₃于50,25和12.5 mg/kg剂量下抑制率分别为95.8%,70.3%和44.2%,它与布洛芬效力(25 mg/bg抑制率72.9%)相当.Ⅱ类化合物体外对L1210细胞及体内对荷艾氏腹水癌小鼠均有一定抗癌活性。     

丁基苯酞对原代培养的大鼠皮层神经细胞外液6-keto-PGF1α和TXB2及其比值的影响

目的旨在观察丁基苯酞(NBP)对神经细胞培养液中6-酮-PGF_1α和TXB₂含量及其比值的影响。用放射免疫方法,结果发现神经细胞在低糖低氧5h或低糖低氧5h/恢复糖氧3h条件下,d-,l-和dl-NBP(0.1～100μmol·L^-1)能够剂量依赖性升高细胞外液中的6-酮-PGF_1α含量,降低TXB₂水平,从而使6-酮-PGF_1α与TXB₂比值升高。而阿司匹林仅在小剂量(0.1,1μmol·L^-1)时能升高6-酮 PGF^1α与TXB^{2比值,大剂量(10,100μmol·L-1)时无影响。提示:NBP对6-酮-PGF_1α/TXB₂比值的升高可能与其增加局部脑血流和改善缺血性脑损伤有关。}     

丁基苯酞对低糖低氧诱导的大鼠皮层神经细胞损伤的保护作用

为进一步探讨丁基苯肽对神经的保护作用,用原代培养大鼠皮层神经细胞的方法,以LDH和细胞形态等指标,观察了l-丁基苯酞(l-NBP)和d-丁基苯酞(d-NBP)对低糖低氧诱导的大鼠皮层神经细胞损伤的保护作用。结果表明:l-NBP和d-NBP能剂量依赖性地抑制低糖低氧诱导的大鼠皮层神经细胞内LDH的释放,降低细胞死亡率,并能改善受损细胞的形态;此外还能明显减轻低糖低氧诱导的神经细胞内粗面内质网脱颗粒及多聚核糖体解聚。提示:NBP对低糖低氧诱导的大鼠皮层神经细胞损伤有明显保护作用。     

抗疟新药的研究5-取代苯氧基-6-甲氧基-8-取代氨基喹啉的合成

根据5-(对-氟苯氧基)-6-甲氧基-8-(4-氨基-1-甲基-丁氨基)喹啉(Ⅰ₁)(表1)对猴疟原虫(Plasmodium cynomolgi)的作用略优于伯喹而毒性较低的报道,合成了化合物Ⅰ₁(代号M7844),同时还合成了衍生物5-取代苯氧基-6-甲氧基-8-(4-取代氨基-1-甲基丁氨基)喹啉(Ⅰ_3～17)(表1)以及其同分异构体5-取代苯氧基6-甲氧基-8-(5-取代氨基戊氨基)喹啉(Ⅱ_13～28)(表2)。药理研究证明化合物Ⅰ_1～7、Ⅰ₁₆及Ⅱ₁₈等对鼠疟P.yoelii均有不同程度的作用。化合物Ⅰ₁(M7844)的毒性甚低,对小鼠的毒性比磷酸伯喹低20余倍,对家兔的溶血反应也明显低干伯喹。但在相同剂量下,对猴疟P.cynomolgi的作用不及伯喹。对鼠疟红前期的作用比伯喹低4～5倍。     

消旋17-苯-18,19,20-失三碳前列腺素F2α甲酯及其15-差向异构体的合成

从中间体(4)出发,经醛(5),与鏻叶立德(3)或2-酮-4-苯丁烷磷酸酯(11)钠缩合成(6),再钠硼氢还原得3′α-醇(7A)及其差向异构体(7B),经硅胶柱色谱分开,分别经二异丁基铝氢还原,与溴化5-三苯鏻戊酸之Wittig试剂缩合,得17-苯-18,19,20-失三碳前列腺素F_2α(9A)及其15-差向异构体(9B),再用重氮甲烷甲酯化,分别得相应的17-苯-18,19,20-失三碳前列腺素F_2α甲酯(10A)及其15-差向异构体(10B)。     

N-(4-甲氧羰基-4-邻苯二甲酰亚氨基丁酰基)-N-取代甘氨酸、脯氨酸和焦谷氨酸的合成

报道11个预期有血管紧张素转化酶抑制活性的N-(4-甲氧羰基-4-邻苯二甲酰亚氨基丁酰基)-N-取代甘氨酸(VII_1～9)、脯氨酸(VII₁₀)和焦谷氨酸(VIl₁₁)的合成和鉴定。所有上述化合物以及与VⅡ_1～9相应的叔丁酯(VI_1～9)均未见文献报道。药理初试结果显示，化合物VII₈，VII₉和VI₁₀均有明显降压活性。     

丁基苯酞对大脑中动脉阻断后皮层组织中花生四烯酸释放及磷脂酶A2基因表达的影响

目的　研究丁基苯酞（dl-NBP), d-NBP和 l-NBP对大脑中动脉阻断(MCAO) 6 h后缺血区皮层中花生四烯酸(AA)释放及磷脂酶A₂（PLA₂）基因表达的影响。方法　阻断大脑中动脉起始部造成局灶性脑缺血模型。HPLC检测AA。Northern blot检测皮层中PLA₂基因表达。结果　MCAO后6 h,皮层中AA释放明显增加。于脑缺血后5 min 和120 min,给dl-NBP(10或20 mg.kg^-1) 和尼莫地平(0.5 mg.kg^-1) 可显著抑制AA的释放。d-NBP和l-NBP作用比较,显示d-NBP有与dl-NBP相似的作用,而l-NBP则无明显影响。Northern印迹结果表明,脑缺血6 h,皮层中PLA₂的基因表达增强。 dl-NBP和d-NBP(10, 20 mg.kg^-1,ip)皆可使表达降低,而l-NBP对缺血脑组织中PLA₂的基因表达的升高无明显影响。结论　dl-NBP和d-NBP可抑制MCAO后脑组织中AA释放和PLA₂的基因表达。     

Synthesis of tritium labelled (R) and (S)-3-aminoquinuclidine: A ubiquitous component of serotonin receptor ligands,part II

(S)-3-Aminoquinuclidine-³H 10c-S having a specific activity of 66 Ci/mmol was prepared in eight steps from Isonicotinic acid ( 2 ). Reduction of 2 with carrier free tritium gas over PtO₂ in DMF gave isonipecotic acid-³H 3c . Conversion to α-bromo ketone 5c followed by cyclization afforded 3-quinuclidone-³H 6c . Racemic 3-aminoquinuclidine-³H 8c was prepared by conversion of 6c to oxime 7c followed by reduction with NaBH₄/NiCl₂·6H₂O. Racemic 8c was resolved with (R)-methylbenzyl isocyanate. Hydrolysis of 9c-S.R afforded (S)-3-aminoquinuclidine-³H 10c-S . The enantiopurity was >99.5% (S).     

A SOLID PHASE-FRAGMENT APPROACH TO PEPTIDE SYNTHESIS.

The t-alkyloxycarbonylhydrazide-resin, H₂NNHCOOC(CH₃)₂CH₂CH₂-copolystyrene-divinylbenzene, was applied to the solid phase synthesis of a protected decapeptide hydrazide, Z-Ala-Val-Ser(Bzl)-Ile-GIn-Phe-Met(O)-Asn-Leu-NHNH₂, and the t-alkyl alcohol-resin, HO-C(CH₃)₂CH₂CH₂-copolystyrene-divinylbenzene, was applied to the synthesis of Z-Lys(Z)-Phe-Phe-Gly-OH. The protected tetrapeptide was coupled to H-Leu-Met-OCH₃ by a fragment condensation in solution to give the protected hexapeptide, which was converted by ammonolysis and deprotection with hot trifluoroacetic acid to the active analog of eledoisin, H-Lys-Phe-Phe-Gly-Leu-Met-NH₂     

Analysis of the Interactions of Cytochrome b5 with Flavocytochrome P450 BM3 and its Domains

Interactions between a soluble form of microsomal cytochrome b₅ (b₅) from Musca domestica (housefly) and Bacillus megaterium flavocytochrome P450 BM3 and its component reductase (CPR), heme (P450) and FAD/NADPH-binding (FAD) domains were analyzed by a combination of steady-state and stopped-flow kinetics methods, and optical spectroscopy techniques. The high affinity binding of b₅ to P450 BM3 induced a low-spin to high-spin transition in the P450 heme iron (K_d for b₅ binding?=?0.44 μM and 0.72 μM for the heme domain and intact flavocytochrome, respectively). The b₅ had modest inhibitory effects on steady-state turnover of P450 BM3 with fatty acids, and the ferrous-carbon monoxy P450 complex was substantially stabilized on binding b₅. Single turnover reduction of b₅ by BM3 using stopped-flow absorption spectroscopy (k_lim?=?116 s^?1) was substantially faster than steady-state reduction of b₅ by P450 BM3 (or its CPR and FAD domains), indicating rate-limiting step(s) other than BM3 flavin-to-b₅ heme electron transfer in the steady-state reaction. Steady-state b₅ reduction by P450 BM3 was considerably accelerated at high ionic strength. Pre-reduction of P450 BM3 by NADPH decreased the k_lim for b₅ reduction ～10-fold, and also resulted in a lag phase in steady-state b₅ reduction that was likely due to BM3 conformational perturbations sensitive to the reduction state of the flavocytochrome. Ferrous b₅ could not reduce the ferric P450 BM3 heme domain under anaerobic conditions, consistent with heme iron reduction potentials of the two proteins. However, rapid oxidation of both hemoproteins occurred on aeration of the ferrous protein mixture (and despite the much slower autoxidation rate of b₅ in isolation), consistent with electron transfer occurring from b₅ to the oxyferrous P450 BM3 in the complex. The results demonstrate that strong interactions occur between a eukaryotic b₅ and a model prokaryotic P450. Binding of b₅ perturbs BM3 heme iron spin-state equilibrium, as is seen in many physiologically relevant b₅ interactions with eukaryotic P450s. These results are consistent with the conservation of structure of P450s (particularly at the heme proximal face) between prokaryotes and eukaryotes, and may point to as yet undiscovered roles for b₅-like proteins in the control of activities of certain prokaryotic P450s.     

Binding of3H-desipramine to the neuronal noradrenaline carrier of rat phaeochromocytoma cells (PC-12 cells)

Summary The specific (i.e., nisoxetine-sensitive) binding of³H-desipramine was studied in purified plasma membranes of PC-12 cells (rat phaeochromocytoma cells).³H-desipramine bound reversibly and with high affinity (K _D=4.5 nmol/l) to a single, non-interacting site (Hill coefficient=1.04); the maximal number of binding sites (B _max) was 19.6 pmol/mg protein.Like the uptake of noradrenaline (by uptake₁), the binding of³H-desipramine was dependent on both sodium and chloride. The stimulation of binding by chloride and sodium was characterized by a Hill coefficient of about 1 and 2, respectively. Both, chloride and sodium, slowed the rate of dissociation of bound³H-desipramine. Increasing concentrations of sodium decreased theK _D of³H-desipramine binding without altering theB _max.The binding of³H-desipramine was inhibited by tricyclic antidepressants and other noradrenaline uptake blockers. There was a highly significant correlation between the potencies of a series of drugs for the inhibition of³H-desipramine binding and for the inhibition of³H-noradrenanne uptake into intact PC-12 cells. Both, binding of³H-desipramine and uptake of³H-noradrenaline, were stereoselectively inhibited by the enantiomers of cocaine and oxaprotiline. However, for most of the substrates of uptake₁ the IC₅₀ for inhibition of³H-desipramine binding was much higher than that for inhibition of³H-noradrenaline uptake. Nevertheless, noradrenaline competitively inhibited³H-desipramine binding and unmasked dissociation of bound³H-desipramine. Thus,³H-desipramine probably binds to the substrate recognition site.From theB _max of³H-desipramine binding to PC-12 membranes and from theV _max of³H-noradrenaline uptake into PC-12 cells, a duration of about 400 ms for the transport cycle for a single noradrenaline molecule was calculated. In addition, from theB _max the maximum number of³H-desipramine binding sites (carriers) for a single PC-12 cell was calculated to be 55,000.A part of this study was presented at the IUPAHR 9th International Congress of Pharmacology (Bönisch et al. 1984)This study was supported by the Deutsche Forschungsgemeinschaft (Bo 521 and SFB 176)     

芳氧乙基季銨盐的制备

为了寄生虫病的实驗治疗,制备了一系列β-芳氧乙基季銨盐。带有各种不同取代基团的苯酚与二甲基或二乙基β-氯乙基胺在丙酮溶液中于无水碳酸鉀的作用下縮合,成为相应β-芳氧乙基二甲基或二乙基胺,后者也可由各該苯酚先与过量溴化乙烯縮合成1-芳氧基-2-溴乙烷,再与二甲胺或二乙胺作用制成。这些胺化合物再与碘甲烷、溴化苄或溴化对硝基苄共热,便成相应β-芳氧乙基季銨盐。对硝基苯基β-溴乙基硫醚与三甲胺縮合,生成对硝基苯巯乙基三甲基季銨盐。     

Efficient solution phase synthesis and use of multiple O-phosphothreonyl-containing peptides for calcium phosphate binding studies

The protected phosphothreonine derivative Boc-Thr(PO₃Ph₂)-OH was prepared in high yield from Boc-Thr-OH by a simple three-step procedure which involved 4-nitrobenzylcarboxyl protection, either phosphorotriester (diphenyl phosphorochloridate) or “phosphite-triester” (diphenyl N.N-diethylphosphoramidite) phosphorylation of the thrconine hydroxyl group of Boc-Thr-ONb followed by hydrogenolytic carboxyl deprotection. The three Thr(P)-containing peptides, H-Thr(P)-Glu-Glu-NHMe.TFA, H-Thr(P)-Thr(P)-Glu-Glu-NHMe.TFA and H-Thr(P)-Thr(P)-Thr(P)-Glu-Glu-NHMe.TFA, were prepared in high yield by the use of Boc-Thr(PO₃Ph₂)-OH in the Boc mode of peptide synthesis (mixed anhydride method) followed by platinum-mediated hydrogenolytic deprotection of the Thr(PO₃3Ph₂)-containing peptides. The use of the phosphopeptides in calcium phosphate binding studies showed that the triple Thr(P)-cluster was a basic structural requirement, since only the pentapeptide was able to bind calcium phosphate efficiently.     

Application of Box-Behnken design in understanding the quality of genistein self-nanoemulsified drug delivery systems and optimizing its formulation

The purpose of the present study was to understand the effect of formulation variables of self- nanoemulsified drug delivery systems (SNEDDS) on the rapid dissolution of a model drug, genistein (GN). A three-factor, three-level Box-Behnken design was used to explore the main and interaction effect of several independent formulation variables including the amount of Maisine 35-1 and Labrafac Lipophile WL 1349 (1:1, w/w) (X₁), Cremophor EL and Labrasol (3:1, w/w) (X₂), and Transcutol P (X₃). Droplet size (Y₁), turbidity (Y₂), and dissolution percentage of GN after 5 (Y₃) and 30 (Y₄) min were the dependent variables. A mathematical relationship, Y₃?=???89.3447?+?5.9524X₁?+?1.0683X₂?+?0.462X₃???0.0825X₁²???0.0075X₂²???0.0009X₃²?+?0.0104X₁X₂ ??0.0113X₁X₃?+?0.0009X₂X₃ (r2?=?0.9604), was obtained to explain the effect of all factors and their co-linearities on the dissolution of GN at 5?min. Formulation optimization was then performed to maximize dissolution percentage of GN at 5?min (Y₃). The optimized formulation was predicted to dissolution 93.34% of GN at 5?min, when X₁, X₂ and X₃ values were 37.1, 101.7 and 77.3?mg, respectively. A new batch was prepared according to the optimized formulation, and the observed and predicted values of Y₃ were in close agreement. In conclusion, the Box-Behnken experimental design allowed us to understand the effect of formulation variables on the rapid dissolution of GN from SNEDDS, and optimize the formulation to obtain a rapid drug dissolution at 5?min.     

Structure-activity studies on C-terminal hirudin peptides containing sulfated tyrosine residues

To clarify the role of the negative charge of the C-terminal region of hirudin, we chemically synthesized the C-terminal peptide of hirudin variant-1 (HV-1), HV-1-(54-65), and its analogs, [E61Y,E62Y]HV-1-(54-65) and [E62Y]HV-1-(54-65), and then sulfated the Tyr residue(s) in these peptides by both enzymic and chemical methods. Enzymic O-sulfation of Tyr residues in the peptides by use of sulfotransferase isolated from Eubacterium A-44 allowed us to produce four kinds of the sulfated peptide, whose C-terminal sequences were -PEY(SO₃H)YLQ, -PYY(SO₃H)YLQ, -PYYY(SO₃H)LQ and -PYY(SO₃H)Y(SO₃H)LQ. On the other hand, all Tyr residues in the peptides were successfully sulfated by chemical reaction with N, N′-dicyclohexylcarbodiimide in the presence of sulfuric acid. Based on the analysis of structure-activity relationships of these sulfated peptides for thrombin inhibition, the Tyr62 and Tyr63 bisulfated peptide GDFEEIPEY(SO₃H)Y(SO₃H)LQ was found to be the most potent inhibitor of thrombin among the products tested. No increase in potency was observed by further substitution of Glu61 with Tyr(SO₃H). The inhibitory activity by substitution with Tyr(SO₃H) at position 63 was greater than that obtained by the substitution at position 62. © Munksgaard 1996.     

The stereoselective κ-opioid receptor antagonist Mr 2266 does not exhibit stereoselectivity as an antagonist at the orphan opioid (ORL1) receptor

Mr 2266 [(-)-(1R,5R,9R)-5,9-diethyl-2-(3-furylmethyl)-2’-hydroxy-6,7-benzomorphan] is an antagonist at κ-opioid receptors and at ORL₁ receptors as well. The aim of our study was to examine whether the known stereoselective antagonism of Mr 2266 at κ-opioid receptors also extends to ORL₁ receptors. In mouse brain cortex membranes, the binding of the ORL₁ receptor agonist [³H]nociceptin was equipotently inhibited by Mr 2266 and its enantiomer Mr 2267 (pK _i 4.82 and 5.14, respectively), whereas the binding of the κ-opioid receptor agonist [³H]U-69,593 was inhibited by Mr 2266 more potently (pK _i 9.11) than by its enantiomer Mr 2267 (pK _i 7.15). In mouse brain cortex slices preincubated with [³H]noradrenaline, the concentration-response curve of nociceptin for inhibition of the electrically evoked overflow of tritium was equipotently shifted to the right by Mr 2266 and Mr 2267 (pA ₂ 5.77 and 5.64, respectively). On the other hand, the inhibitory effect of U-69,593 on the electrically evoked overflow of tritium in guinea-pig brain cortex slices preincubated with [³H]noradrenaline was more potently antagonized by Mr 2266 (pA ₂ 8.81) than by Mr 2267 (pA ₂ 7.15). These data show that the stereoselective antagonism of Mr 2266 at κ-opioid receptors does not extend to ORL₁ receptors. Received: 15 October 1998 / Accepted: 12 November 1998     

Synthesis,toxicity, and percutaneous activity of silicon glycerolates and related hydrogels

Silicon glycerolates of the general formula Si(C₃H₇O₃)₄ ⋅ xC₃H₈O₃(3 ≤ x ≤ 10) and related hydrogels of the general formula Si(C₃H₇O₃)₄ ⋅ xC₃H₈O₃ ≤ yH₂O (3 ≤ x ≤ 10, 20 ≤ y ≤ 40) were synthesized and some of their pharmacological properties were studied. The laws of gel formation were investigated and optimum conditions for the process were determined. High percutaneous (transdermal) activity of the synthesized compounds was revealed by measuring the diffusion of drugs through intact skin membrane in vitro. The acute and chronic toxicity was studied. It was established that all substances are nontoxic. The experimental results show that silicon glycerolates and related hydrogels can be recommended for further testing and investigation as effective biologically active percutaneous vehicles of medicinal preparations. Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 42, No. 11, pp. 5–9, November, 2008.