环孢菌素A产生菌发酵条件的优化研究

利用“单因素多浓度”和“均匀设计法”对环抱菌素生产菌株茄病镰刀菌的发酵条件和发酵培养基组分进行优化．并在5000L发酵罐上对菌株的发酵动力学规律作了初步的探索。实验结果表明。5．5％玉米粉,0．5％葡萄糖,0．5％干酪素．0．05％KCl,0．05％MgSO4,0．015％KH2PO4和0．3％CaCO3为最佳摇瓶发酵培养基组成;5000L发酵罐的发酵动力学规律表明,菌体浓度发酵培养至80h左右达到最大．产素水平在116h左右达到最大。     

环孢菌素A生物合成机制研究的新进展

环孢菌素A(CsA)是一种环状+一肽化合物(图1),已广泛地用临床人体器官移植和治疗自身免疫疾病。 CsA是由雪白白僵菌(Beauveria　nivea)产生的。至今已鉴定了25种天然产生的CsA组份,它们或在1、2、4、5、7和11位氨基酸置换,或在1、4、6、9、10和11位含未甲基化的肽键。此外,通过产生菌发酵时添加氨基酸前体,获得与CsA在1、2和8位有不同氨基酸的新的环孢菌素组份。     

环孢菌素A两种不同产生菌原生质体的制备和再生

以环孢菌素A两种不同产生菌——雪白白僵菌和茄病镰刀菌为出发菌株，开展了原生质体制备和再生条件的研究，并分别实现了两个不同种的菌株原生质体再生。     

小剂量环孢菌素A治疗类风湿关节炎

目的观察小剂量环孢菌素A(CsA)治疗活动性或难治性类风湿关节炎(RA)的疗效及其安全性. 方法 18例RA患者给予CsA 2mg*kg-1*d-1,分 2次口服,3个月为一疗程.治疗前,治疗4周、8周与治疗3个月时,测定反映关节功能各项指标,血沉、C反应蛋白、类风湿因子(RF)定量、肝肾功能,治疗前及治疗3个月后摄关节X线片,治疗1周后及第6周测定CsA血浓度.结果有效率4周为66.0%,8周和3个月分别为7 8.0%和83.3%, 显效5例、有效10例、无效3例,疗程结束后血沉及RF定量明显下降,C反应蛋白无明显变化.测定CsA的血浓度在40～90μg/L,无一例出现CsA的毒副作用.结论 CsA治疗4周左右即可显示疗效,8周左右可见具有统计学意义的临床变化,40～90μg/L的血浓度可能是治疗RA安全有效的窗口治疗浓度.     

环孢菌素A血药浓度分析方法进展

环孢菌素A是一种高效免疫抑制剂,在器官、组织移植及治疗自身免疫疾病方面应用广泛.由于其药代动力学及药效学特点,对其进行血药浓度检测意义重大,多种血药浓度分析方法进展较快,但目前临床仍以高效液相色谱法及荧光偏振免疫法为多数.     

环孢菌素A的临床应用概况

环孢菌素 A(CyclosporinA CsA)又称环孢菌素,是一种新型强效免疫抑制剂。目前本药除作为器官移植的首选药物外,其它方面的临床应用也日趋广泛,现概述如下。1 移植物抗宿主病 此病系指器官及骨髓移植后对宿主的排斥反应,CsA对其有良好的防治作用。罗永礼等在肾移植术当天及术后第1天予以CsA(4～5mg/kg)静滴,以后改为口服维持治疗1年。结果:人/肾1年存活率88.6％/81.8％,排斥率20.9％,细菌感染率     

环孢菌素A的临床应用

环孢菌素A除用于器官移植外,近年来广泛用于肾病综合征、狼疮性肾炎、溃疡性结肠炎、再生障碍性贫血等自身免疫性疾病,本文就其临床应用作简要综述。 1 器官移植 环孢菌素A能防止器官移植时排斥反应和移植物抗宿主反应,由于环孢菌素的应用,脏器移植的成功率大幅度提高。1年生成率为:活体肾移植从70％上升到90％;尸体肾移植从40％～50％上升到80％;心移植从60％上升到80％,肝移植从40％上升到70％～80％,从而使脏器移植成为正规的临床治疗方法。1987年美国共进行肾移植约万次,心移植约1500多次,肝移植约1100多次。据罗永礼等报道,我国对202例206次尸体肾移植术后服用环     

果葡糖浆对环孢菌素A发酵影响的研究

目的 研究F42果葡糖浆对环孢菌素A发酵的影响.方法 离心检测菌丝体浓度及HPLC法检测发酵过程中环孢菌素A发酵单位.结果 F42果葡糖浆在去除部分葡萄糖后更加有利于环孢菌素A发酵,菌丝浓度和产量明显增加,确定最适浓度为11%,其发酵单位与与8%果糖组相当,而菌丝浓度明显低于8%果糖组.结论 由于去除部分葡萄糖的F42果葡糖浆价格远低于果糖,在生产中可以用于替代果糖,从而降低生产成本.     

氨基酸对Tolypocladium inflatum产生环孢菌素A的影响

环孢菌素A(Cy A)是一种具有免疫抑制及抗真菌活性的环状十一肽。使其闻名的主要专利系在人体移植外科和控制自体免疫病方面的用途。环孢菌素A是由丝状真菌Tolypocladium inftatum在深层发酵中产生。抗生素分子上的氨基酸组分可引导和促进真菌的肤及缩肽抗生素的产生。Kobel等报道了发酵中CyA和几种类似物的定向合成,而各种外加的氨基酸强烈决定着所产生的每种类似物的组成与效价。这可能是由于环孢菌素合成酶缺乏专一性,它是参与次级代谢产物特别是肽类抗生素非核糖体合成的多功能合成酶的通性。     

环孢菌素A制剂的研究进展

目的介绍环孢菌素A的制剂研究进展。方法综述近年来国内外相关文献,介绍环孢菌素A的作用机制、临床应用及制剂研究,提出环孢菌素A制剂的研究前景。结果环孢菌素A新制剂不仅可以提高环孢菌素A的生物利用度、降低其毒性和不良反应,同时可扩大其临床应用范围。结论环孢菌素A新制剂的研究为环孢菌素A进一步发挥免疫抑制和其他作用提供了新途径。     

The dose-dependent inhibition of rat renal translation elongation seen after in vivo cyclosporin A is not caused by cyclosporin metabolites

Cyclosporin A (CsA) given to Sprague-Dawley rats in vivo produced a tissue-specific, dose-dependent inhibition of translation elongation in renal microsomes. CsA at an oral dose of 50 mg/kg/day for 6 days reduced renal microsomal translation by 70.5%. Renal cytoplasm from rats treated in vivo with CsA inhibited translation by 55% when added to renal microsomes isolated from tissues of control animals. In contrast, CsA added to renal microsomes in vitro did not inhibit translation. Renal cytoplasm from CsA-treated rats containing translation inhibitory factor was found by HPLC to contain CsA and CsA metabolites M1 and M17. CsA metabolites M1, M17, M18 and M21 were isolated from human bile and tested in vitro for translation elongation inhibitory activity in renal microsomes. CsA, M18 and M21 did not inhibit translation elongation at concentrations of up to 2500 ng/ml. M17 inhibited translation elongation, but only by 8.4% at the highest concentration tested (2500 ng/ml), a level 20-fold higher than that measured in renal cytoplasm (125 ng/ml). Ml produced a concentration-dependent inhibition of translation elongation, beginning at 500 ng/ml, or approximately 2-fold higher than that found in renal cytoplasm (260 ng/ml). M1 at 2500 ng/ml or approximately 10-fold higher than the concentration measured in renal cytoplasm, inhibited translation elongation by 23.8%, only 1/3 that observed upon addition of renal cytoplasm containing translation inhibitory factor. We conclude from these findings that the dose-dependent inhibition of renal translation elongation following in vivo CsA cannot be explained by the renal formation or uptake of known CsA metabolites.     

反相高效液相色谱法测定全血中环孢素A的浓度

目的：建立检测患者全血中环孢素A（CsA）浓度的高效液相色谱法。方法：血样经乙醚-石油醚（70：30）萃取后,以C18硅胶键合相为固定相,乙腈-甲醇（80：20）为流动相,70℃柱温下进行色谱分析,紫外检测波长为210nm。结果：CsA全血浓度在50-1600ng／mL范围内呈良好的线性关系,r=0．9998,方法的平均回收率为97．6％,日内和日间精密度RSD均小于6％（n=5）,最低检测限为10ng／mL。结论：本法简便、快速,易于开展,用于肾移植术后患者CsA的血药浓度监测,效果良好。     

环孢素A对实验大鼠肺纤维化的干预作用

目的 研究环孢素A(CsA)对大鼠肺纤维化的干预作用.方法 平阳霉素(BLM)气管内注入建立大鼠肺纤维化模型,观察CsA对支气管肺泡灌洗液(BALF)中细胞成分和转化生长因子-β(TGF-β)的含量、肺组织学变化,应用SABC法检测肺组织中纤维连接蛋白(FN)和血小板衍化生长因子(PDGF)表达的作用.结果 应用BLM后第7天即有严重肺泡炎性改变,第28天有严重的肺纤维化,而CsA组,早期只有轻度炎性改变,BALF中细胞计数及TGF-β含量较BLM组低(P＜0.05),FN及PDGF表达显著弱于BLM组(P＜0.01).结论 CsA可能通过抑制FN、PDGF的表达及TGF-β的生成干预BLM致肺纤维化.     

反相高效液相色谱法测定肾移植术后全血环孢素A浓度

目的建立单步萃取反相高效液相色谱法测定人全血中环孢素A(CsA)浓度。方法患者血样经乙醚单步萃取,环孢素D(CsD)为内标,C18色谱柱,乙腈-水(80:20)为流动相,流速1.0ml/min,柱温70℃,检测波长214nm。结果CsA全血浓度在50～600ng/ml范围内线性关系良好,r=0.9998,最低检测浓度为20ng/ml,平均回收率100.44%,日内和日间RSD均低于5%。结论反相高效液相色谱法简便灵敏、准确可靠、经济,可用于临床人体器官移植术后全血中CsA浓度的监测。     

Experimental pulmonary delivery of cyclosporin A by liposome aerosol

The utilization of CsA–liposome for aerosol delivery by jet nebulizers has potential advantages for clinical development including: aqueous compatibility, sustained pulmonary release to maintain therapeutic drug levels and facilitated delivery to alveolar macrophages and pulmonary lymphocytes. Inhalation of cyclosporin A (CsA)–dilauroylphosphatidylcholine (DLPC) liposome aerosols will theoretically result in localized and sustained delivery of therapeutic CsA concentrations within the lung as an alternative to local immunotherapy for pulmonary diseases. In the lung, targeted delivery of therapeutic CsA concentrations would require lower dosages than via conventional intravenous or oral routes of administration. Potential benefits from targeted lung delivery could include reduced systemic toxicity and prolonged immunosuppressive activity. Aerosol delivery systems have been developed to deposit drugs directly onto pulmonary surfaces at the sites of disease within the lung. A novel HPLC method for tissue analysis of CsA–liposomes is developed and utilized with a solid-phase extraction method to measure CsA recovered from Balb/c mouse lung tissues. A concentrated formulation containing 5 mg CsA–37.5 mg DLPC/ml was nebulized with an Aerotech II nebulizer generating an aerosol particle size distribution (mass median aerodynamic diameter (MMAD)) of 1.7 μm and geometric standard deviation (GSD) of 2.0. After a 15-min aerosol exposure, little of no CsA was detected in the blood, liver, kidney or spleen. The lung contained the highest organ CsA levels with high immunosuppressive activity demonstrating effective pulmonary targeting of the CsA–DLPC liposome aerosol. The results of this system will be utilized as the experimental basis for future pharmacokinetic, toxicological, immunosuppression and other biological studies.     

康宁克通-A与小剂量环孢菌素A联合治疗口腔扁平苔癣的疗效观察

目的 :观察局部注射皮质类固醇激素 (康宁克勇—A)与口服小剂量环孢菌素A联合治疗口腔扁平苔癣 (OLP)的临床效果。方法 :对58例OLP患者进行联合治疗 (联合治疗组 ) ,对54例进行单纯局部治疗 (对照组 )。对照组采用康宁克通—A(40mg/ml) ,抽取0 5～1ml ,加等量2 %利多卡因 ,在病损区粘膜下作多点和基底注射 ,每周1次 ,4周为1疗程。联合治疗组在采用康宁克通—A的同时加服环孢菌素A(25mg/片 ) ,每日1片 ,4周为1疗程。观察4个疗程的痊愈率 ,随访3年 ,观察痊愈者的复发率并作比较。结果 :采用联合治疗的病例其痊愈率显著高于单纯治疗者 (P<0 05) ;联合治疗的复发率则显著低于单纯治疗 (P<0 01)。结论 :局部激素封闭注射联合口服小剂量环孢菌素A治疗OLP可提高痊愈率 ,减少复发率。     

环孢素A微乳浓缩液的药代动力学和生物等效性评价

将雄性Ｗｉｓｔａｒ大鼠１６只随机分为两组,分别口服自制和进口环孢素Ａ（ＣｙＡ）微乳浓缩液,采用高效液相色谱法测定血药浓度,对其药代动力学和相对生物利用度进行了研究。试验结果表明两种制剂中ＣｙＡ的药动学过程均符合口服吸收二室模型。自制和进口环孢素Ａ微乳浓缩液的全血药物浓度达峰时间分别为１．５７±０．５５和１．６８±０．４３ｈ;Ｃｍ ａｘ 分别为１７５５．６±２２６．０和１８３２．２±５９８．８ｎｇ／ｍｌ;Ｔ１／２ 分别为１９．９３±６．４４和１９．７９±６．９８ｈ;ＡＵＣ分别为３０６３７．９±７５５２．４和３０３１６．６±６５７８．９ｎｇ·ｈ／ｍ ｌ;自制环孢素Ａ微乳浓缩液的相对生物利用度为１０１．１％ 。经统计分析,两种制剂的药代动力学参数均无显著性差异（Ｐ＞ ０．０５）,两种制剂具有生物等效性。     

Inhibition of T cell cAMP formation by cyclosporin A and FK506

The influence of the immunosuppressants, cyclosporin A (CsA) and FK506, on cAMP formation was studied in T cells from healthy controls and patients with psoriasis. While basal cAMP levels were not affected, CsA (1 M) and FK506 (2 nM) prevented the isoprenaline (0.1 M)-induced increase in cAMP formation. Half-maximal inhibition by FK506 and CsA was observed at about 0.2 nM and 20 nM, respectively. In addition, both agents significantly reduced (by about 50%) the forskolin (8 M)-stimulated cAMP formation. No differences were noted in CAMP responses (basal, stimulation by isoprenaline, inhibition by CsA and FK506) of T cells from healthy controls and psoriatic patients. We conclude that CsA and FK506 potently and efficiently interfere with the stimulatory adenylyl cyclase pathway in T cells and that regulation of T cell CAMP formation is apparently not altered in psoriasis.This paper contains data of the doctoral thesis of P. Oeljeklaus. Parts of this work were presented at the Annual Meeting of the Society for Investigative Dermatology, Washington, DC, USA, April 1994     

Dose dependent absorption and linear disposition of cyclosporin A in rat

The pharmacokinetics of cyclosporin A (CyA) were investigated in the rat following intravenous doses of 1·7, 3·3, and 6·4 mg kg^?1 and oral doses of 3·1, 6·8, and 12·9 mg kg^?1. The blood concentration-time profile after intravenous administration was adequately described by a two-compartment model when all data were simultaneously analysed using NONMEM. The disposition pharmacokinetics were linear over the dose range studied; the average total blood clearance was 0·191 g^?1 kg^?1. The absorption process could not be adequately described by either a first- or a zero-order input. Therefore, a flexible, staircase input model was used and found to be superior to the standard models. The shape of this model was biphasic, with a higher initial input rate than expected from first-order absorption. The duration of this first phase increased with dose. The extent of absorption was also dose dependent. Bioavailability was higher at higher doses; the values were 45%, 67% and 76% for the three ascending dose levels. These results strongly indicate a saturable first-pass effect. Since the extraction of CyA in the liver is only 6%, the marked increase in bioavailability of CyA is most likely to be the result of saturated gut wall metabolism.