真菌来源白细胞介素6受体拮抗剂2520A的研究

目的从真菌Torulomyces ovatus的代谢产物中分离白细胞介素6受体(IL-6R)拮抗剂。方法利用多种柱色谱进行分离纯化对IL-6R具拮抗活性的物质，根据理化性质、光谱数据确定其结构。结果从真菌Torulomyces ovatus的代谢产物中得到对IL-6R有拮抗活性的化合物2520A。结论2520A化合物为已知铁螯合物ferrichrome，首次报道该化合物具IL-6R拮抗活性，并确定其结构含有铁原子。     

白细胞介素-1受体拮抗剂致畸敏感期毒性的试验

白细胞介素-1(IL-1Ra)属于炎症细胞因子,可促进炎症及组织损伤的病理效应。白细胞介素-1受体拮抗剂(IL-1Ra)则是一种存在于人体的蛋白质类细胞因子受体拮抗剂,能特异性地封闭IL-1的活性,具有很好的临床应用前景。根据中华人民共和国卫生部《新药(西药)临床前研究指导原则汇编》     

白细胞介素-1受体拮抗剂与气道高反应性关系的研究

目的 :探讨白细胞介素 1受体拮抗剂 (IL 1ra)与哮喘的气道高反应性 (AHR)的关系。方法 :正常豚鼠 ,先腹腔注射卵清蛋白 (OA)致敏而后雾化吸入OA激发豚鼠哮喘 (哮喘组 ) ,激发前 30min静脉注射IL 1ra 1mg·kg-1(哮喘 IL 1ra组 ) ,测定豚鼠的气道反应性 (组胺的PC2 0 值 ) ,并收集支气管肺泡灌洗液 (BALF)进行细胞总数计数及分类记数。结果 :哮喘 IL 1ra组PC2 0 值与对照组无显著性差异 (P >0 .0 5 ) ,显著高于哮喘组 (P <0 .0 1) ;BALF中的中性粒细胞 (NES)的百分比显著低于哮喘组 (P <0 .0 1)。所有豚鼠均存在 IgPC2 0 值与BALF中的NES的百分比呈正相关 (r =0 .95 ,P <0 .0 0 1)。结论 :IL 1ra能降低AHR ,其机制可能与IL 1ra减少NES的浸润有关     

白细胞介素1受体拮抗剂对胰腺炎大鼠胰腺细胞凋亡的诱导

目的 从胰腺腺泡细胞凋亡的角度探讨白细胞介素1受体拮抗剂(IL-1Ra)对急性胰腺炎腺泡细胞的诱导.方法 用牛黄胆酸钠建立大鼠急性坏死性胰腺炎模型,IL-1Ra腹腔注射诱导胰腺腺泡细胞凋亡,并应用细胞凋亡原位标记检测(TUNEL)染色,检测胰腺组织腺泡细胞的凋亡情况.结果 HE染色见胰腺组织中典型的细胞核固缩及凋亡小体形成.IL-1Ra干预后1、2、6、12 h胰腺细胞凋亡指数显著高于非治疗组相应时间点(P＜0.01).结论 IL-1Ra治疗急性胰腺炎的机制不仅限于对炎症介质的抑制,且能够通过诱导胰腺腺泡细胞的凋亡减轻胰腺炎的严重程度.     

白细胞介素1受体拮抗剂对佐剂性关节炎大鼠免疫功能的影响

采用大鼠佐剂性关节炎模型，在致炎后ｄ１８取出关节炎大鼠脾脏和腹腔渗出液，制备脾细胞和腹腔巨噬细胞悬液，并运用３Ｈ－ＴｄＲ参入法等免疫学方法观察白细胞介素１受体拮抗剂（ＩＬ－１ｒａ）对佐剂性关节炎大鼠的淋巴细胞增殖功能、ＩＬ－２和ＩＬ－１产生的影响。结果表明，ＩＬ－１ｒａ（１～３２μｇ·Ｌ－１）在体外能明显抑制大鼠腹腔巨噬细胞产生ＩＬ－１，促进脾淋巴细胞增殖及ＩＬ－２的产生。可能与其作用于白细胞介素１受体有关。     

重组人白细胞介素-1受体拮抗药对豚鼠离体呼吸道平滑肌的影响

目的　观察人重组白细胞介素 1受体拮抗剂 (IL 1ra)对正常和卵白蛋白致敏豚鼠离体肺条、气管平滑肌的影响。方法　应用离体器官装置、张力换能器、MedLab记录系统测定肺条和气管平滑肌的张力。结果　①IL 1ra对正常豚鼠离体肺条和气管平滑肌有直接松弛作用 ,EC50 分别为1 2 9ⅹ 10 -7mol·L-1和 8 0 6× 10 -8mol·L-1;并对卵白蛋白致敏的豚鼠肺条和气管平滑肌也有直接的松弛作用 ,EC50 分别为 2 6 1× 10 -7mol·L-1和 5 88× 10 -7mol·L-1,但致敏豚鼠呼吸道平滑肌对IL 1ra的敏感性要比正常豚鼠低。②IL 1ra(10 -9～ 10 -5mol·L-1)可剂量依赖性地抑制致痉剂组胺对正常豚鼠肺条和气管平滑肌的收缩作用 (P <0 0 1)。③IL 1ra能抑制卵白蛋白攻击引起的肺条和气管平滑肌的收缩 ,IC50 分别为 7 83ⅹ 10 -7mol·L-1和 4 4 8ⅹ 10 -7mol·L-1。结论　IL 1ra对正常、痉挛及致敏状态的呼吸道平滑肌均有松弛作用     

白细胞介素-6小分子拮抗剂的研究进展

白细胞介素6(IL6)是一种多功能细胞因子,在免疫应答、造血调控和急性期反应中起重要作用。然而,IL6的异常表达与许多自身免疫性疾病和肿瘤的发病机制及发展进程相关。因此,发展针对IL6的拮抗剂将有可能用于这些疾病的治疗。近年来,已有部分IL6拮抗剂用于临床,但多为中和抗体等大分子。由于这些大分子存在相对分子质量大、不易保存、抗原性强等缺点,所以发展新型特异性拮抗IL6的小分子药物将成为治疗IL6表达异常相关疾病的新途径。本文就IL6小分子拮抗剂的研究进行初步探讨。     

白细胞介素1受体拮抗剂筛选模型研究

以链霉菌表达的重组ｈｓＩＬ－１ＲＩ为靶位,建立了基于受体配基竞争性结合为原理、白细胞介素１受体拮抗剂筛选模型,对５００株微生物发酵产物进行了筛选,复筛阳性率为４％～５％,该模型具有新颖性和实用性。     

重组人白细胞介素-1受体拮抗剂对大鼠胶原性关节炎的治疗作用

目的　研究重组人白细胞介素 1受体拮抗剂 (recom binanthumaninterleukin1receptorantagonist, rhIL 1ra)对大鼠胶原性关节炎 (collagen inducedarthritis,CIA)的治疗作用。方法　SD大鼠随机分为 6组:正常对照组、模型组、rhIL 1ra3个剂量组和进口IL 1ra组;采用Ⅱ型胶原(CⅡ)乳剂皮内注射诱导大鼠CIA模型;每周测体重观察体重变化;足爪容积法测量大鼠足爪肿胀度;测定CⅡ的迟发性变态反应(DTH);ELISA法检测血清中抗Ⅱ型胶原抗体水平;同时进行病理组织学的检查。结果　CIA大鼠在致炎d10,出现关节红、肿,体重减轻,血清抗CⅡ抗体水平升高,关节病理可见滑膜增生,炎性细胞浸润,血管翳形成,软骨和骨的破坏等。rhIL 1ra(7 5, 30, 120mg·kg-1 )和阳性对照组anakinra(120mg·kg-1 )皮下注射,连续 7d,能明显抑制CIA大鼠足肿胀;rhIL 1ra(30, 120mg·kg-1 )皮下注射,连续 7d,可以明显减轻CⅡ诱发的迟发性变态反应 (DTH);另外,rhIL 1ra也可明显降低CIA大鼠血清中抗CⅡ抗体的水平;病理学检查表明,rhIL 1ra大剂量能明显减轻CIA大鼠关节滑膜炎症和滑膜增生,减轻血管翳和软骨破坏。结论　rhIL 1ra对CIA具有治疗作用。     

白细胞介素-1受体阻滞剂滴眼液耐受性研究

目的:评价白细胞介素-1受体阻滞剂(IL-1Ra)滴眼液在中国健康人的安全性和耐受性。方法:依据其临床前安全资料确定初剂量及最大剂量,评价其安全性。采用单次给药、连续多次给药的方法,观察用药后生命体征、眼局部安全指标和全身安全性指标。结果:IL-IRa的耐受性试验未引起受试者全身、局部症状的异常,未引发不良反应症状。结论:IL-1Ra滴眼液在25～100μg·mL-1范围内,人体耐受性良好。推荐进行Ⅱ期临床试验的用药剂量及疗程为50μg·mL-1滴眼液qid,每次0．05 mL,共用药5 d。     

微生物来源白细胞介素1受体拮抗剂的研究Ⅱ．链霉菌660代谢产物的化学研究

从活性链霉菌660的发酵液中,经溶媒萃取,硅胶制备薄层层析及ODS等分离得到化合物4和化合物6,通过理化性质研究和UV、IR、MS、^1H-NMR、^13C-NMR、DEPT、^1H-^1H COSY、^1H-^13C COSY、HMBC等光谱分析确定化合物4为N－乙酰基酪胺（N-acetyltyramine),化合物6为3－丙基－7－甲基－六氢吡咯[1,2-a]并吡嗪－1,4－二酮（3－propyl-7-methyl-hexahydropyrrolo[1,2-a]pyrazine-1,4-dione)。并对其活性进行研究。     

Studies on the biochemical actions of 6-selenoguanine and 6-selenoguanosine

Survival studies were performed in mice bearing Sarcoma 180 ascites tumor treated with 6-thio and 6-seleno analogs of guanine and guanosine. The selenium-containing analogs were somewhat superior to the sulfur-containing compounds in antitumor activity and therapeutic index. The formation of 6-SeGMP from 6-seleno-guanine (6-SeG) was demonstrated in Sarcoma 180 ascites cells. Guanine, 6-thioguanine (6-TG) and 6-SeG show comparable substrate activity whereas 8-azaguanine is a much poorer substrate for hypoxanthine-guanine phosphoribosyl transferase from Sarcoma 180 cells. Both 6-TG and 6-SeG are good substrates for purine nucleoside phosphorylase from Sarcoma 180 cells. Chemically and enzymatically synthesized 6-SeGMP behaved as a competitive inhibitor (K_i 1 × 10^?4 M) of erythrocytic and Sarcoma 180 guanylate kinases. Weak substrate activity was demonstrated in the presence of large amounts of erythrocytic guanylate kinase.     

IL-6及IL-6R和Survivin在胃癌组织中的表达及意义

目的:探讨胃癌组织中的白细胞介素6(IL-6)及其受体(IL-6R)、survivin的表达及其相互关系。方法:应用原位杂交技术检测86例胃癌标本中IL-6及其受体mRNA的表达;应用免疫组织化学方法(S-P法)检测86例胃癌变标本中survivin蛋白的表达。结果:胃癌组织中Survivin蛋白、IL-6及IL-6R阳性表达率分别为58.1%(50/86)、74.4%(64/86)、73.3%(63/86)。Survivin蛋白与IL-6及其受体的表达与胃癌的组织分化、浸润深度、淋巴结转移密切相关(P<0.05),但与患者的性别、年龄差异均无显著性(P>0.05);Survivin的表达与IL-6呈正相关(P<0.01)。结论:IL-6及IL-6R和Survivin是反应胃癌发生发展的较好指标;具有抑制胃癌细胞凋亡的作用。     

IL-6R distribution in normal human and cynomolgus monkey tissues

Interleukin-6 (IL-6) is a pleiotropic cytokine and a contributing factor in many diseases such as rheumatoid arthritis, Castleman’s disease, Crohn’s disease, and multiple myeloma. Since the blockade of the signaling pathway of the IL-6/interleukin-6 receptor (IL-6R)/gp130 complex is considered to have therapeutic value in such diseases, we developed an IL-6R humanized antibody (tocilizumab). In the current report, distribution of IL-6R in both normal human and cynomolgus monkey tissues was assessed as fundamental data to support preclinical and clinical studies of tocilizumab. Human and cynomolgus monkey tissue panels were stained with commercially available anti-human IL-6R and a species- and isotype-matched negative antibody, as well as assay control slides. The detection system applied used an Envision^® immunoperoxidase staining procedure with DAB reaction. Positive reactions were observed in the tissue elements of lymphatic, hematopoietic, digestive, reproductive, exocrine, endocrine, neural, muscular, epidermal, respiratory, and urinary systems of the human and cynomolgus monkey tissue panels. The current report is inclusive of a wide variety of tissues and shows the distribution of IL-6R to be similar for both human and monkey tissues. We consider this information fundamental for the support and interpretation of preclinical and clinical studies of anti-IL-6R antibody therapy.     

The IL-6/sIL-6R complex as a novel target for therapeutic approaches

IL-6 plays a pivotal role in immune responses and certain oncologic conditions. The intense investigation of its biological activity and function led to the discovery of two different IL-6-driven signalling pathways. Binding to the membrane-bound IL-6 receptor (mIL-6R, CD126) causes the recruitment of two gp130 co-receptor molecules (CD130) and the activation of intracellular signalling cascades via gp130. Although this classical pathway is mainly limited to hepatocytes, neutrophils, monocytes/macrophages and certain other leukocyte populations, which express IL-6R on their surface, an alternative mechanism has also been described. Proteolytic cleavage of the mIL-6R protein or translation from alternatively spliced mRNA leads to the generation of a soluble form of the IL-6R (sIL-6R), which is likewise able to bind to IL-6. The resulting IL-6/sIL-6R complex is also capable of binding to gp130 and inducing intracellular signalling. Through this so-called ‘trans-signalling’ mechanism, IL-6 is able to stimulate cells that lack an endogenous mIL-6R. High levels of IL-6 and sIL-6R have been reported in several chronic inflammatory and autoimmune diseases as well as in cancer. Preclinical animal disease models have provided strong evidence that specific blockade of IL-6-regulated signalling pathways represents a promising approach for the therapy of these diseases. An optimised variant of the recently described fusion protein sgp30Fc is now heading towards its clinical evaluation.     

5-HT6 receptor antagonists enhance retention of a water maze task in the rat

RATIONALE: 5-HT(6) receptors are predominantly located in the brain and may be involved in cognitive processes. The aim of this study was to assess the effects of two potent and selective 5-HT(6) receptor antagonists, SB-271046-A and SB-357134-A, on learning and memory in the rat. METHODS: Spatial learning and memory was assessed by testing the effects of SB-271046-A and SB-357134-A on acquisition and retention of a water maze task. RESULTS: In the water maze, administration of SB-271046-A or SB-357134-A (3 or 10 mg/kg) had no effect on learning per se. At 10 mg/kg, however, both compounds produced a significant improvement in retention of a previously learned platform position when tested 7 days after training. By contrast, the acetylcholinesterase inhibitor, Aricept (donepezil, 0.1, 0.3 mg/kg PO) had no effect in this task. CONCLUSIONS: This study demonstrates that systemic administration of SB-271046-A and SB-357134-A produces improvements in retention of a water maze task in the rat. These data indicate that 5-HT(6) receptor antagonism may be involved in cognitive function.     

海洋真菌09-1-1-1次级代谢产物的研究

目的从海洋真菌的代谢产物中寻找新的具有抗真菌活性的化合物。方法以稻瘟霉生物模型筛选海洋真菌,获得编号09-1-1-1的活性菌株,从其发酵液中分离活性代谢产物。结果从发酵液中分离到2个化合物,鉴定其结构为3a,12c-二氢-8-羟基-6-甲氧基-7H-呋喃[3′,2′∶4,5]呋喃[2,3-c]呫吨-7-酮(Ⅰ,sterig-matocystin)和1,3,6,8-四羟基-2-(1-羟己基)-9,10-蒽二酮(Ⅱ,averantin)。结论稻瘟霉生物模型用于筛选海洋真菌活性代谢产物成本低、快速又方便。化合物Ⅱ对109稻瘟霉菌丝的生长最小抑制浓度为1.6μg.mL-1。     

益督丸对老龄小鼠白细胞介素—2和白细胞介素—6活性的影响

目的:研究益督丸的免疫调节作用。方法:用放射免疫法(RIA)测定白细胞介素2(IL2)的活性,用KD83细胞株MTT法测定白细胞介素6(IL6)的活性。结果:益督丸对老龄小鼠IL2活性有明显升高作用。而对老龄小鼠异常升高的IL6水平则有降低作用。结论:益督丸的补肾、益督、壮阳等作用与调节机体免疫功能有关。     

阿托伐他汀治疗对急性冠脉综合征患者血浆CRP和IL-6水平的影响

目的探讨阿托伐他汀干预对急性冠脉综合征（ACS）患者血浆C反-应蛋白（CRP）和白介素-6（IL-6）水平的影响。方法采用随机开放法将62例ACS分为阿托伐他汀治疗组和常规对照组进行为期4周的干预,分别测定治疗前及治疗后2周患者血浆CRP和IL-6的浓度变化,比较治疗前后的血浆CRP及IL-6水平变化。结果急性冠脉综合征患者血清CRP及IL-6水平明显升高,治疗4周后,阿托伐他汀治疗组血浆CRP及IL-6水平均明显下降,而常规对照组CRP和IL-6水平无明显变化。结论ACS的发生与机体炎症反应激活有关。阿托伐他汀能够降低血浆CRP和IL-6水平,在ACS的治疗中具有重要作用。     

IL-10抑制脂多糖诱导的Hela细胞IL-15和IL-6转录

目的研究IL-10对脂多糖(LPS)诱导的Hela细胞IL-15mRNA和IL-6 mRNA表达的影响,分析其激活的信号转导通路。方法培养的Hela细胞,经不同浓度的LPS及IL-10单独或联合处理后,提取细胞总RNA和总蛋白,RT-PCR分析IL-15和IL-6转录水平的变化,Westernblot分析信号转导通路蛋白变化。结果RT-PCR分析得知①1ng～10μgLPS刺激Hela细胞12h后,IL-15 mRNA和IL-6 mRNA水平均明显上调(与对照组比较:P<0.01),且存在剂量依赖关系,100ng/mL时达峰值;100ng/mLLPS刺激Hela细胞0～24h,IL-15 mRNA和IL-6 mRNA水平亦明显上调(与对照组比较:P<0.01),24h内存在时间依赖关系,12h时达峰值。②单独IL-10(10ng/mL)作用于Hela细胞12h后,IL-15 mRNA和IL-6 mRNA没有明显变化(与对照组比较:P>0.05)。不同浓度的IL-10(1,10,100ng/mL)均下调100ng/mLLPS诱导的Hela细胞IL-15 mRNA和IL-6 mRNA的表达,且浓度越高IL-10抑制作用越明显。Western blot分析显示LPS主要通过磷酸化信号蛋白PI3K/AKT和ERK1/2上调IL-15和IL-6转录,IL-10能阻断AKT的磷酸化而对ERK1/2的磷酸化没有影响。结论IL-10可抑制LPS诱导的炎症细胞因子IL-15和IL-6的转录,这可能与其阻断AKT的磷酸化有关,因而,IL-10可能应用于某些临床感染性疾病的预防和治疗。