重组人血小板生成因子注射液的Ⅰ期临床耐受性试验

目的:研究国产重组人血小板生成因子注射液(rhTPO)在健康志愿者体内的耐受性和安全性。方法:随机选择健康志愿者27例,分别单次皮下注射rhTPO 0．5,1．0,2．0μg·kg-1,进行耐受性研究,动态观测健康志愿者用药后血象、血小板形态、凝血酶原时间和血清生化指标等。结果:除1例受试者用药36 h后四肢出现皮疹外,其余未观察到明显毒性反应;血小板计数约于给药后14 d达高峰,给药后21 d基本回落至基础水平,血小板升高时形态、功能及凝血酶原时间无明显改变。结论:单次皮下注射rhTPO 0．5～2．0μg·kg-1对人体是安全的,不良反应轻微,具有特异性和剂量依赖性升高血小板作用,对血小板形态和功能无明显影响。推荐Ⅱ期临床给药剂量为皮下注射1．0μg·kg-1,qd,连续7 d。     

重组链激酶治疗幼猪急性脑梗死的药效学

目的 :研究重组链激酶 (rSK)治疗急性脑梗死的药效学。方法 :幼猪 72头 ,分 7个静脉组 ,3个动脉组及 2个对照组 ,每组各 6头。幼猪脑梗死4h后分别经静脉和颈内动脉 (ICA)应用rSK ,动态检测ICA血流量及脑血管造影。 2 4h后取脑作病理检查。结果 :静脉组rSK 2万IU·kg- 1或以上剂量的血管再通率大于对照组 (差异有显著意义 ,P <0 .0 5 ) ;同一剂量动脉组再通率大于静脉组 (P >0 .0 5 )。血管完全再通 ,部分再通及不通的病理学检查脑梗死发生率分别为 7% ,5 0 %和 10 0 % (差异有显著意义 ,P <0 .0 5 )。结论 :静脉用rSK治疗急性脑梗死的最低有效剂量为 2万IU·kg- 1;局部动脉给药以 1～ 2万IU·kg- 1为宜。     

磷酯酶C抗血小板功能的研究Ⅰ——对血小板聚集率和粘附率的作用

目的　通过研究PLC对家兔血小板粘附率和聚集率的作用 ,以探讨PLC对血小板功能的影响。方法　家兔麻醉后 ,经十二指肠给予阴性对照辅料 ,阳性对照ASA和 6种剂量的PLC ,于给药前及给药后 1、2、4h颈动脉取血测定以ADP、AA、Collagen为诱导剂时的血小板聚集率 ,同时用玻球法测定血小板的粘附率。结果　PLC 10 0、 2 0 0IU·kg-1对家兔血小板粘附率无明显影响。PLC 4 0 0～ 10 0 0IU·kg-1可显著地降低血小板粘附率 ,并呈现一定的剂量效应依赖关系。同一剂量PLC组给药后 1、2、4h时相比P <0 0 5。PLC10 0～ 10 0 0IU·kg-16种剂量组在用ADP、AA、Collagen诱导时 ,均可显著地抑制血小板聚集 ;与阴性对照辅料组相比P <0 0 5 ,与阳性对照组相比 ,在ADP诱导时 ,10 0IU·kg-1组的抑制较弱 (P <0 0 5 ) ,而 80 0、10 0 0IU·kg-1组的抑制较强 (P <0 0 5 ) ,并呈一定的剂量效应和时间效应关系 ;而在AA诱导时 ,PLC各组在给药后 2、4h时的抑制率与ASA的相当 ,无明显剂量效应依赖关系 ,而有一定的时间效应关系 ;在Collagen诱导时 ,PLC各组的抑制作用均弱于ASA(P<0 0 5 ) ,无明显的剂量效应依赖关系 ,而有一定的时间效应关系。结论　①PLC经十二指肠给药 ,10 0、2 0 0IU·kg-1对家兔血小板粘附性无明显影响 ,而 4     

五步蛇毒纤溶组分Ⅱ对大鼠颈动脉血栓的溶栓作用

目的了解五步蛇毒纤溶组分Ⅱ对大鼠颈动脉血栓有无溶栓作用。方法采用大鼠颈动脉血栓模型，舌下静脉注射五步蛇毒纤溶组分Ⅱ，分为3个剂量组（n=10），其剂量分别为625μ·kg-1、1250μVg·kg-1和2500μ·kg-1。用尿激酶作用阳性对照，其剂量分别为1250IU·kg-1、2500IU·kg-1和5000IU·kg-1。用生理盐水作阴性对照。结果给药lh后，五步蛇毒纤溶组分Ⅱ组溶解血栓的重量（三上s）分别是（1.9士0.6）mg、（4.8土0.9）mg和（7.0土0.8）mg。尿激酶组溶解血栓的重量（王士s）分别是（l·6上0·5）mg、（2.3土0.7）mg和（3.0土0.4）mg。生理盐水组溶栓重量（Z士s）为（0.6土0.2）mg。五步蛇毒纤溶组分正组、尿激酶组与生理盐水组作t值检验，PMO.01。结论五步蛇毒纤溶组分I和尿激酶对大鼠颈动脉血栓都有溶栓作用，并有量效关系。     

聚乙二醇胸腺素α1注射液的Ⅰ期临床耐受性研究

目的:观察聚乙二醇胸腺素α1注射液在健康志愿者的安全性和耐受性。方法:单次给药剂量组的36名健康志愿者分别接受7个剂量组(0.16,0.8,1.6,3.2,4.8,6.4,9.6 mg)的聚乙二醇胸腺素α1注射液,多次给药剂量组的16名健康志愿者分别接受2个剂量组(3.2和4.8 mg,每周1次,连续给药4周)的聚乙二醇胸腺素α1注射液,观察受试者的生命体征、实验室检查、心电图检查结果的变化,记录试验期间发生的不良事件。结果:单次给药试验中,3.2 mg组发生1例ALT升高的不良事件,程度为中度,可能与药物有关;9.6 mg组发生4例注射部位硬结的不良事件,判断为皮下注射不完全吸收所致。多次给药试验中,4.8 mg组发生1例第1次给药后头晕、心悸的不良事件,退出试验,与受试者主观因素有关,与药物无关。其余受试者给药前后的生命体征、实验室检查、心电图检查均未见有临床意义的改变。结论:聚乙二醇胸腺素α1注射液单次给药0.16~6.4 mg和连续4周,3.2~4.8 mg,每周1次,中国健康志愿者安全耐受。     

国产重组人血小板生成素单次皮下注射的药代动力学研究

目的研究国产重组人血小板生成素单次皮下注射在人体内的药代动力学特征.方法24名健康志愿者随机分为0.5 mg.kg-1,1.0 mg.kg-1和2.0 mg.kg-13个剂量组,单次皮下注射相应剂量的重组人血小板生成素.于给药前和给药后不同时间取血,分离血清,采用ELISA测定血药浓度.结果正常人皮下注射重组人血小板生成素0.5,1.0和2.0 μg.kg-1后,tpeak分别为9.0±1.9 h,10.8±2.4 h和11.8±5.1 h,Cmax分别为0.298±0.081 mg.L-1,0.438±0.076 mg.L-1和0.831±0.079 μg.L-1,AUC与给药剂量基本成正比.人体内重组人血小板生成素的消除较缓慢,3个组的t1/2ke分别为46.3±6.9 h,40.2±9.4h和38.7±11.9 h.结论当以0.5～2.0μg.     

复方银花解毒颗粒对干酵母诱导幼鼠发热模型的退热作用研究

目的：研究复方银花解毒颗粒对干酵母所致幼龄大鼠发热模型的解热作用,为复方银花解毒颗粒的儿童给药剂量提供参考。方法：将SPF级3-5周龄幼年雄性SD大鼠分为正常对照组、模型组、布洛芬组、复方银花解毒颗粒低中高三个剂量组（给药剂量分别为2.5、5、10 g·kg-1）。各组灌胃给药,连续7天,末次给药1 h后皮下注射干酵母混悬液制备发热模型,造模后于不同时间点分别测定肛温。检测血清中IL-1β、IL-6、TNF-α含量以及下丘脑内cAMP水平。Western blot检测下丘脑炎症信号相关COX2的含量,并检测NF-κB的核转录情况,检测相关IκBα磷酸化水平变化。结果：造模后2~4 h,布洛芬、复方银花解毒颗粒高剂量组（10 g·kg-1）均能抑制发热大鼠体温升高,在3 h时体温降低效果显著;造模3h后,与模型组比较,复方银花解毒颗粒低剂量组（2.5 g·kg-1）能显著降低IL-6、IL-1β水平;复方银花解毒颗粒中剂量组（5 g·kg-1）、高剂量组（10 g·kg-1）能显著降低IL-6、TNF-α含量。复方银花解毒颗粒高剂量组（10 g·kg-1）下丘脑内NF-κB、IκBα、COX2蛋白表达量与模型组相比也显著下降。结论：复方银花解毒颗粒能有效降低发热模型幼鼠的体温,降低体内炎症因子水平,其退热机制可能与cAMP信号转导通路和NFκB信号通路有关。     

重组人类促红细胞生成素防治早产儿贫血的临床研究

目的　应用不同剂量国产重组人类促红细胞生成素 (rhu EPO)防治早产儿贫血 ,探讨其最适剂量和疗效。方法　 83例孕周 <36周 ,出生体重 <2 5 0 0克早产儿 ,以入院顺序按rhu EPO剂量随机分为第 1组 2 5例 (rhu EPO 75 0IU·kg-1·w-1) ,第 2组 18例 (45 0IU·kg-1·w-1) ,第 3组 2 0例(30 0IU·kg-1·w-1) ,分为每周 3次 ,静脉注射 ,疗程 4周。另设第 4组 2 0例为对照组。结果　 4组早产儿生后血红蛋白 (Hb)、红细胞压积 (Hct)渐行下降。第 1组下降最轻 ,第 2组次之 ,对照组最明显 ,经方差分析有显著性差异 (P <0 0 1) ;网织红细胞 (Ret)明显增高 ,且Ret升高程度和rhu EPO剂量有关 ;治疗后血清EPO浓度以第 1组最高 ,对照组最低 ,有显著性差异 (P <0 0 1) ,但第 3组与对照组比较无显著性差异 (P >0 0 5 ) ;血清铁 (Fe)生后均逐渐下降 ,治疗各组略低于对照组 ,无显著性差异。结论　早期rhu EPO治疗可提高Hb、Hct、Ret,并且疗效和剂量有关 ,体内充足的铁储备是确保rhu EPO疗效的重要因素。     

重组人胰高血糖素样多肽-1(7-36)在健康人体的药代动力学

目的研究重组人胰高血糖素类多肽-1(7-36)[rhGLP-1(7-36)](促胰岛素生成药)在健康志愿者的药代动力学。方法选择12,9名健康志愿者,分别单次(0.1,0.15,0.2mg)和多次(0.2mg)皮下注射rhGLP-1(7-36),连续5日,用荧光酶联免疫分析法测定其血药浓度,用DAS软件计算其药代动力学参数。结果单次:药-时曲线符合一房室模型,Cmax、AUC0-t随剂量增加而增加,tmax约为19~21min,t1/2为10~14min。多次:首次与末次给药后的Cmax分别为(726.76±94.07),(737.15±72.12)ng·L-1;tmax为18min左右;t1/2为15~17min。结论在0.1~0.2mg内,过程呈一级线性动力学特征,体内无蓄积,耐受性较好。     

20(R)-人参皂苷Rg3人体药代动力学研究

目的　研究 2 0 (R) 人参皂苷Rg3(GRg3)人体药代动力学。 方法　高效液相色谱 -紫外检测法。结果8名健康志愿者单剂量口服 3 2mg·kg-1GRg3 ,其药时曲线符合口服吸收有滞后时间的二房室模型 ,Tmax为 (0 6 6± 0 10 )h ,Cmax为 (16± 6 )ng·mL-1,T1/ 2α为 (0 46± 0 12 )h ,T1/ 2 β为 (4 9± 1 1)h ,T1/ 2 (Ka) 为 (0 2 8± 0 0 4)h ,AUC0 -∞ 为 (77± 2 6 )ng·mL-1·h ;6名健康志愿者单剂量口服 0 8mg·kg-1GRg3 ,由于血药浓度低 ,可测数据点少 ,未进行模型模拟 ;两组给药剂量与相应Cmax实测值比较 ,二者成正比关系。结论　本品口服吸收快 ,消除也较快 ,但血药浓度很低。在所试剂量范围内 ,GRg3属一级动力学吸收、消除过程。     

Effect of human kidney lipids on human kidney renin activity

The major lipids of human kidney tissue were isolated by solvent extraction, and the lipid composition was determined by thin-layer chromatographic techniques. The positional distribution of fatty acyl groups in ethanolamine and choline phosphatides was determined after enzymatic hydrolysis. Major phosphatides were assayed for plasmalogen content. Triglycerides were characterized by argentation chromatography. The fatty acyl composition of these lipids was also determined. The effect of intact triglycerides, phospholipids, 1- and 2-monoacyl phosphatides and ether lipids on renin activity in vitro was determined by incubations with 3-[U¹⁴C]valyl tetradecapeptide renin substrate. Kidney triglycerides, 1-monoacyl and 2-monoacyl phosphatidylethanolamines and phosphatidylcholines significantly inhibited renin activity. The renin-inhibitory effect of these lipids was comparable to inhibition by hog kidney phospholipid inhibitor. The intact phospholipids and cholesterol potentiated human kidney renin activity. Phosphatidylserines and synthetic glyceryl ether lipids have no significant effect. These results indicate that lipid-induced inhibition of human renin activity does not require the ethanolamine moiety, acyl group unsaturation, or the presence of a hydroxyl group at the 2-position. Additionally, no specific structure-activity relationships can describe lipid-renin interactions.     

Priming effect of recombinant human interleukin-2 and recombinant human interferon-gamma on human neutrophil superoxide production

The effect of recombinant human interleukin-2 (rhIL-2) and recombinant human interferon-gamma (rhIFN-gamma) were evaluated on superoxide (O2-) production of human polymorphonuclear neutrophils (PMNs). Ten minutes incubation with rhIL-2 showed a dose-dependent enhancement of n-formyl-methionyl-leucyl-phenylalanine (FMLP)-induced O2-production of human PMNs, and the rate of enhancement reached 49.6% at the concentration of 3000 U/ml rhIL-2. Same pretreatment with rhIFN-gamma also showed a dose-dependent enhancement of FMLP-induced O2-production of human PMNs, and the maximal rate of enhancement was 47.0% at the concentration of 3000 U/ml rhIFN-gamma. Any cytokines given alone did not induce O2- production. These cytokines showed no enhancement of phorbol myristate acetate (PMA)-induced O2- production, neither. The effects of these cytokines to FMLP-induced O2- production were kept in calcium-free medium. Moreover, incubation with these cytokines caused no elevation of intracellular free calcium concentration [( Ca++]i) in resting PMNs. Incubation with them did not change the increase of [( Ca++]i) of PMNs induced by FMLP significantly, neither. Recombinant forms of cytokines are used clinically, now. These results may be helpful for the use of them.     

Antitumor effects of recombinant human prolactin in human adenocarcinoma-bearing SCID mice with human NK cell xenograft

To survey the immune regulatory function of recombinant human prolactin (rhPRL) and its potential application in adoptive immunotherapy, CB17-SCID mice were loaded with human colon adenocarcinoma HT-29 cells (5 x 10(5) cells/mouse, i.p.) 24 h before adoptive transfer with the purified human NK cells followed by rhPRL injection (10 mug/mouse, every other day for a total of 10 injections). Upon analysis, rhPRL did not exert any direct inhibitory effects on HT-29 cells but slightly improved the tumor cell growth both in vitro and in vivo. After SCID mice were reconstituted with human NK cells, rhPRL improved the antitumor effects of human NK cells in HT-29-bearing SCID mice, showing a prolonged survival from 70.4 to 112.1 days, and the increased survival rate from all died to 40% survival for more than 160 days. rhPRL improved the proliferation of human NK cells with or without PHA stimulation. rhPRL also directly enhanced the cytotoxicity of human NK cells against HT-29 tumor cells in 4-h coculture. The supernatant of rhPRL-stimulating NK cells inhibited the proliferation of HT-29 cells through, at least partly, the interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) in the supernatant. Thus, rhPRL administration in HT-29 tumor-bearing SCID mice promotes the antitumor effects of adoptively transferred NK cells.     

Effect of human plasma on the reactivation of sarin-inhibited human erythrocyte acetylcholinesterase

The reactivation of organophosphate-inhibited acetylcholinesterase (AChE) by oximes inevitably results in the formation of highly reactive phosphoryloximes (POX), which are able to re-inhibit the enzyme. In this study, the dependence of POX formation on AChE concentration was investigated with sarin-inhibited human erythrocyte AChE (EryAChE). A marked dependence was found with obidoxime but not with the experimental oxime HI?6, suggesting great differences in the decomposition rates of the respective POXs. At a physiological erythrocyte content the reactivation of EryAChE was markedly affected by POX with obidoxime and pralidoxime (2-PAM) but not with the newer oximes HI 6 and HLö 7. Addition of extensively dialysed, sarin-treated human plasma reduced the reactivation by obidoxime and 2-PAM even more. Obidoxime and 2-PAM were superior to HI 6 and HLö 7 in reactivating butyrylcholinesterase (BChE). This effect was pronounced in diluted plasma, but was obscured in concentrated plasma, probably because of re-inhibition by the generated POX. Addition of native erythrocytes to sarin-treated plasma resulted in marked inhibition of EryAChE in the presence of obidoxime, suggesting a higher affinity of the POX for EryAChE. The results indicate that obidoxime and 2-PAM may reactivate sarin-inhibited AChE insufficiently due to re-inhibition by the POX formed. In addition, the re-inhibition of EryAChE may be aggravated by the POX that is produced during BChE reactivation. These reactions must be regarded as therapeutically detrimental and disqualify those oximes which are capable of forming stable POX by reactivation of BChE.     

Glucuronidation of olanzapine by cDNA-expressed human UDP-glucuronosyltransferases and human liver microsomes

Olanzapine is a widely used, newer antipsychotic agent, which is metabolized by various pathways: hydroxylation and N-demethylation by cytochrome P450, N-oxidation by flavin monooxygenase and direct glucuronidation. In vivo studies have pointed towards the latter pathway as being of major importance. Accordingly, the glucuronidation reaction was studied in vitro using cDNA-expressed human UDP-glucuronosyltransferase (UGT) enzymes and a pooled human liver microsomal preparation (HLM). Glucuronidated olanzapine was determined by HPLC after acid or enzymatic hydrolysis. The following UGT-isoenzymes were screened for their ability to glucuronidate olanzapine: 1A1, 1A3, 1A4, 1A6, 1A9, 2B7 and 2B15. Only UGT1A4 was able to glucuronidate olanzapine obeying saturation kinetics. The K(m) value was 227 micromol/l (SE 43), i.e. of the same order of magnitude as for other psychotropic drugs, and the V(max) value was 2370 pmol/(min mg) (SE 170). Glucuronidation was also mediated by the HLM preparation, but a saturation level was not reached. The olanzapine glucuronidation reaction was inhibited by several drugs known as substrates for UGT1A4, e.g. amitriptyline, trifluoperazine and lamotrigine. Thus, competition for glucuronidation by UGT1A4 represents a possibility for drug-drug interactions in subjects receiving several of these psychotropic drugs at the same time. Whether such possible interactions are of any clinical importance may await further studies in patients.     

Characterization of fimasartan metabolites in human liver microsomes and human plasma

1.?The metabolites of fimasartan (FMS), a new angiotensin II receptor antagonist, were characterized in human liver microsomes (HLM) and human subjects.

2.?We developed a method for a simultaneous quantitative and qualitative analysis using predictive multiple reaction monitoring information-dependent acquisition-enhanced product ion scanning. To characterize metabolic reactions, FMS metabolites were analyzed using quadrupole-time of flight mass spectrometer in full-scan mode.

3.?The structures of metabolites were confirmed by comparison of chromatographic retention times and mass spectra with those of authentic metabolite standards.

4.?In the cofactor-dependent microsomal metabolism study, the half-lives of FMS were 56.7, 247.9 and 53.3?min in the presence of NADPH, UDPGA and NADPH?+?UDPGA, respectively.

5.?The main metabolic routes in HLM were S-oxidation, oxidative desulfuration, n-butyl hydroxylation and N-glucuronidation.

6.?In humans orally administered with 120?mg FMS daily for 7 days, the prominent metabolites were FMS S-oxide and FMS N-glucuronide in the 0–8-h pooled plasma sample of each subject.

7.?This study characterizes, for the first time, the metabolites of FMS in humans to provide information for its safe use in clinical medicine.     

Phase I comparability of recombinant human albumin and human serum albumin

Recombinant human albumin (rHA) is a highly purified animal-, virus-, and prion-free product developed as an alternative to human serum albumin (HSA), to which it is structurally equivalent. The present investigation compared the safety, tolerability, and pharmacokinetics/pharmacodynamics of rHA with HSA. Two double-blind, randomized trials were performed in healthy volunteers using intramuscular (IM) and intravenous (IV) administration. The IM trial included 500 volunteers, each receiving 5 repeat doses of 5 mg (100 subjects), 15 mg (100 subjects), or 65 mg (300 subjects) of rHA or HSA. Thirty volunteers participated in the IV trial, each receiving ascending doses (10 g, 20 g, and 50 g) of either rHA or HSA. In both trials, all adverse events were recorded and conventionally classified; potential allergic responses were also monitored. Blood samples were taken in both studies to test for IgG or IgE antibodies against test products and potential impurities. For the IV study, pharmacokinetic/pharmacodynamic assessments were performed, including measurement of serum albumin, colloid osmotic pressure, and hematocrit pre- and postinfusion. Nine subjects in the IM study (4 recipients of rHA and 5 of HSA) reported drug-related, potentially allergic events; all but 2 of these were skin related. No serious or potentially allergic events were reported with either product in the IV study. There was no immunological response to either product, and dose level did not influence the study outcomes. Serum albumin, colloid osmotic pressure changes, and hematocrit ratio were as expected, with no differences between rHA and HSA. rHA and HSA exhibited similar safety, tolerability, and pharmacokinetic/pharmacodynamic profiles, with no evidence of any immunological response.