注射用重组人成纤维细胞生长因子-21食蟹猴药动学研究

目的 研究注射用重组人成纤维细胞生长因子-21（FGF-21）在食蟹猴体内单次和多次皮下给药后的药代动力学特征。方法 采用随机、平行和剂量递增设计,18只食蟹猴分别单次sc150、300和600 μg/kg的FGF-21或多次sc 300 μg/kg的FGF-21,按设计采集动态血清样本。采用双抗体夹心ELISA法检测血清中药物浓度的动态变化,采用DAS 3.2.4药动程序拟合并计算药动参数。结果 食蟹猴单次sc FGF-21后,各剂量组峰浓度C_max和药时曲线下面积AUC_（0-24h）、AUC_（0-∞）均随给药剂量的增加而增大,C_max、AUC_（0-24h）和AUC_（0-∞）三者均与剂量呈线性相关,各剂量组t_1/2Z、T_max、CL_z和V_z均较为一致。与单次给药相比,食蟹猴多次sc给药后的t_1/2z有所延长但其他主要PK参数（T_max、V_z和CL_z等）均较为一致,多次给药后,体内药物无蓄积倾向。结论 食蟹猴单次sc FGF-21后,在150~600 μg/kg剂量范围内呈现线性动力学特征。单次与多次sc给予FGF-21后,两者的PK行为特征基本一致,无明显药物蓄积。     

食蟹猴sc重组人干扰素β-1a注射液30 d安全性评价

目的 研究重组人干扰素β-1a （rhIFN β-1a）注射液的毒副反应靶器官及其恢复情况,预测可能引起的临床不良反应,为临床用药提供参考。方法 食蟹猴sc重复给予rhIFN β-1a注射液3.9、13.2、45.0 μg/kg,连续30 d,每组6只动物,雌雄各半,停药后恢复16 d。观察药物对一般体征、饮食、肛温、体质量的影响,同时进行尿液分析、血液学、血液生化、心电图、眼科检查、骨髓涂片、免疫学指标、免疫原性检查,在给药结束和恢复结束进行大体剖检并进行组织病理学检查。结果 45.0 μg/kg rhIFN β-1a组动物出现一过性肛温升高,以及活化部分凝血活酶时间（APTT）、凝血酶时间（TT）延长,血浆纤维蛋白原（FIB）浓度降低,该变化在给药期内能恢复。其余一般指标、心电图、血液生化、尿液生化、骨髓检查、大体解剖、脏器质量及系数、组织学检查、免疫学指标检查未见异常。免疫原性试验可见给药20 d后绝大部分动物开始出现抗体,给药30 d时所有动物均出现抗体,各剂量组绝大多数产生结合抗体的动物血清可检测到具有中和rhIFN β-1a注射液活性的抗体,结合抗体以及中和抗体的滴度均与剂量呈非相关性关系,具有随给药时间延长抗体强度增强的趋势。结论 食蟹猴连续sc重复给予rhIFN β-1a注射液30 d,无可见有害作用水平（NOAEL）为13.2 μg/kg,该剂量为人临床拟用剂量0.135 μg/kg的97.8倍。rhIFN β-1a注射液对食蟹猴具有一定的免疫原性。     

食蟹猴重复给予溶瘤病毒药物HSV-1/hPD-1的毒性研究

目的：考察食蟹猴重复给予溶瘤病毒药物HSV-1/hPD-1后的体内毒性,探索安全剂量范围,为后续临床试验提供参考信息。方法：30只食蟹猴随机分成3组,包括溶媒对照组和低、高剂量 （1.0×10⁸ 、4.0×10⁸ pfu）组,每组10只,雌雄各半。采用肌肉注射给药,每周给药2次,连续给药6周, 恢复期8周。试验期间,每天观察动物的临床症状和摄食量,每次给药后1~2天观察注射部位症状,每周称量体重。分别在检疫期、首次给药后、给药期结束、恢复期结束的不同时间点进行安全药理（体温、 血压、心电图）测定、临床病理（血液学、血凝、血清生化、尿生化）检查、免疫学（T淋巴细胞、细胞因子、免疫原性）测定、组织病理学检查和脏器称重。结果：给药后,动物未见异常症状、注射部位刺激性、体重和摄食量改变,未见安全药理和临床病理指标有意义的变化。与溶媒对照组比较,第41 天,低剂量会引起动物CD3⁺ CD4⁺ T细胞比例升高,高剂量未见明显变化。第13至97天,低、高剂量均能引起动物产生抗载体结合抗体、抗抗体,以及个别动物检出hPD-1表达产物。证明药物在体内产生免疫活性和介导免疫原性。组织病理学检查显示,给药期结束时,低、高剂量组动物注射部位极轻度至中度混合细胞浸润,高剂量组动物坐骨神经极轻度髓鞘/轴突变性;恢复期结束时注射部位病变减为极轻度, 坐骨神经病变未见恢复趋势。低、高剂量组动物未见组织脏器重量改变。结论：食蟹猴重复给予溶瘤病毒药物HSV-1/hPD-1后,动物体内耐受良好,受试物未见毒性反应剂量（NOAEL）是1.0×10⁸ pfu。上述研究结果为药物开展临床试验提供了数据支持。     

注射用重组人成纤维细胞生长因子-21食蟹猴急性毒性试验研究

目的 观察食蟹猴单次sc注射用重组人成纤维细胞生长因子-21（RH-FGF-21）的急性毒性反应。方法 8只健康食蟹猴,随机分成RH-FGF-21组和对照组,RH-FGF-21组6只,对照组2只,均雌雄各半,分别单次sc最大给药浓度和最大给药体积的供试品RH-FGF-21（9.0 mg/kg）和最大体积的对照品灭菌注射用水（1.0 mL/kg）,给药后连续观察21 d。在给药前适应期和给药后观察期内进行动物的一般观察（外观、行为、饮食、反应能力、分泌物和排泄物等）、死亡情况、体质量变化、肛温、进食量、眼科和心电图检查、血液学和血液生化学检测、尿液观察和分析;观察期结束安乐死全部存活动物,进行组织病理学检查。结果 动物体质量、肛温、进食量、心电图检查和眼科检查与同期对照组比较均未见明显异常;血液学检测、血液生化检测、尿液观察和分析结果、组织病理学检查也未见由药物引起的异常变化。结论 在本试验条件下未发现由药物引起的急性毒性症状,9.0 mg/kg可认为是食蟹猴单次sc RH-FGF-21的最大给药量。     

重组抗CD52单克隆抗体的食蟹猴单次静脉注射毒性研究

目的：开展食蟹猴静脉注射重组抗CD52人源化单克隆抗体注射液单次给药毒性研究,考察动物体内耐受性、给药后出现的毒性反应及严重程度和药物毒性作用靶器官,评价其安全性。方法：首先应用流式细胞术检测方法,筛选出8只红细胞表面CD52抗原阴性食蟹猴,用于单次给药毒性研究。将动物随机分成4组,包括溶媒对照组和3、10、30 mg·kg^-1剂量组,每组2只,雌雄各半。采用静脉推注给药,给药1次。试验期间,每天观察动物的临床症状和摄食量,每周称量1次体质量。分别在给药第2、 8、15 d采集动物外周血液进行临床病理检查,包括血液学(含凝血)、血清生化(含电解质)。分别在给药第2、6、8、15、21 d采集动物外周血液进行T淋巴细胞及亚群和B淋巴细胞的分类计数。于给药第 22 d解剖动物并进行大体病理学检查。结果：动物给药后,在10 mg·kg^-1和30 mg·kg^-1剂量下,会引起白细胞总数、淋巴细胞、单核细胞数量和比例、T淋巴细胞数量及其亚群细胞数量下降,这些变化与药物的免疫抑制作用相关。受试物还会引起血红蛋白浓度下降,网织红细胞数量和比例和总胆红素升高。 结论：给予食蟹猴单次静脉注射3、10、30 mg·kg^-1重组抗CD52人源化单克隆抗体注射液后,动物耐受良好,最大耐受量是30 mg·kg^-1。本研究的给药剂量以及与药物相关的毒性发现,为后续开展长期毒性研究提供了参考。     

巴马小型猪植入式遥测动物模型的建立及应用

目的 基于遥测技术建立适用于监测清醒自由活动状态下巴马小型猪心电、血压、体温等生理指标的动物模型,为其在安全药理学研究中的应用提供支持。方法 8只巴马小型猪（雌雄各半）经手术植入植入子,手术后恢复3~4周,经体格检查确认已完全恢复后,采用EMKA遥测系统连续监测至少24 h心电图、血压等生理指标,包括心率、PR间期、QRS间期、QT间期、校正QT （QTcv）间期、体温、收缩压（SBP）、舒张压（DBP）、平均动脉压（MBP）,并于次周重复监测1次。对巴马小型猪的24 h指标变化进行分析,并与Beagle犬和食蟹猴数据进行种属间比对分析。结果 巴马小型猪的心电数据和血压数据昼夜节律均不明显;体温呈现明显的昼夜节律,白天体温高,夜间体温低;动物的心率、血压和体温均受外界影响比较大,与动物的活动呈现明显的相关性;与Beagle犬和食蟹猴比较,巴马小型猪的心率和人类最为相似,SBP仅稍高于人类。结论 通过体内植入植入子,成功建立了巴马小型猪植入式遥测动物模型;与Beagle犬和食蟹猴比较,巴马小型猪更适合用于药物对心血管系统影响的评价。     

氢溴酸樟柳碱注射液大鼠单次和重复给药毒性试验研究

目的 通过SD大鼠的单次和重复静脉给药毒性试验,评价氢溴酸樟柳碱注射液的安全性。方法 单次给药毒性试验采用最大耐受量法,观察大鼠的死亡情况和毒性反应。重复给药毒性试验：将大鼠随机分为溶媒对照组和氢溴酸樟柳碱10、50、200 mg/kg剂量组,每组30只,尾iv给药,连续13周,停药恢复4周。进行各项毒理学指标检测。结果 急性毒性试验：氢溴酸樟柳碱注射液在364.5~504.5 mg/kg对大鼠产生明显毒性,症状有给药时尖叫、俯卧、后肢无力、颤抖、抽搐、惊厥、瞳孔散大、尾部发绀等,甚至造成个别动物死亡。重复给药毒性试验：50、200 mg/kg剂量组出现体质量增长减缓,摄食下降,给药后尖叫,鼻端、眼周异常分泌物增多,皮肤脱毛、结痂,耳廓溃疡、缺损,瞳孔散大,尾部发绀,血红蛋白（HGB）、红细胞压积（HCT）、Cl-浓度升高等症状,200 mg/kg剂量组还出现给药后肌张力减退、颤抖、抽搐、呼吸困难、皮下炎性包块等表现。溶媒对照组和200 mg/kg剂量组动物注射部位均出现静脉炎及静脉周围炎,严重程度无差异,停药4周后病变减轻。结论 氢溴酸樟柳碱注射液SD大鼠静脉单次给药的最大耐受量（MTD）为428.8 mg/kg,约相当于临床剂量的2 573倍;重复给药毒性试验未见明显毒性反应剂量（NOAEL）为10 mg/kg,约相当于临床剂量的60倍。     

精蛋白重组人胰岛素注射液生物类似药非临床安全性评价

目的 通过小鼠单次给药毒性试验、Beagle犬重复毒性及免疫原性试验和豚鼠全身主动过敏试验,考察精蛋白重组人胰岛素注射液（Insulin NPH）的毒副反应、毒性靶器官或靶组织,为开展临床试验提供依据。方法 ①小鼠单次给药毒性试验：采用最大给药量法分别sc生理盐水、溶媒和Insulin NPH（2092~2488 IU/kg）,监测给药后小鼠一般状态、体质量、脏器异常。②Beagle犬重复毒性试验：sc溶媒、原研对照药（Humulin NPH,1.5 IU/kg）,低、中和高剂量（0.5、1.0和1.5IU/kg）的Insulin NPH,每天1次,连续30 d,停药恢复14 d;在给药期和恢复期内观察动物的一般体征和注射部位的局部刺激性,进行体质量、肛温、血糖及心电图检查,测定血液学、血清生化、尿液常规等指标,并进行脏器质量及组织病理学检查;免疫原性试验采用间接ELISA法检测不同给药期Beagle犬血清中抗药结合抗体。③豚鼠主动全身过敏试验：分别sc低和高剂量（4和12 IU/kg）的Insulin NPH、生理盐水和溶媒,另设卵清白蛋白为阳性对照,使用以上剂量进行5次致敏试验后,iv 3倍致敏剂量进行激发试验,观察豚鼠过敏症状。结果 小鼠sc 165倍临床常用剂量的Insulin NPH后,未见明显毒性反应;Beagle犬重复毒性试验中1.0 IU/kg是Insulin NPH的无毒反应剂量（NOAEL）,该剂量相当于临床拟用剂量的2倍,免疫原性试验各剂量组均未发现抗药结合抗体;豚鼠主动全身过敏试验中未见明显过敏症状。结论 在本试验条件下未观察到Insulin NPH明显毒性反应。     

重组三价人乳头瘤病毒疫苗的急性毒性研究

目的 评价重组三价人乳头瘤病毒（HPV）疫苗的急性毒性。方法 将Wistar大鼠随机分成2组：阴性对照组和HPV疫苗组,每组20只,HPV疫苗组单次sc 3倍人用剂量（1.5 mL）HPV疫苗,阴性对照组注射等体积的生理盐水。观察动物临床状态;检测动物体质量变化;给药后第15天,解剖大鼠,进行大体病理学检查。结果 动物单次给药大剂量HPV疫苗后,临床症状均未见明显异常;与对照组比较,给药组动物体质量增长未见显著性差异;病理学检查未见与供试品相关的明显异常。结论 Wistar大鼠单次sc重组三价HPV疫苗总体耐受性良好,未见明显毒性反应,为进一步临床前评价HPV疫苗的安全性奠定了基础。     

治疗用乙型肝炎腺病毒注射液TC007食蟹猴免疫原性、免疫毒性和生物分布研究

目的 重复给予食蟹猴治疗用乙型肝炎腺病毒注射液（TC007）,观察其免疫原性、免疫毒性、生物分布。方法 普通级食蟹猴随机分为溶媒对照组,TC007低、中、高剂量（1.0×10⁹、1.0×10¹⁰、1.0×10¹¹ VP/只）组,Ad5-Null（1.0×10¹¹ VP/只,空载对照）组,每组10只,背部肩胛区sc给药,每周给药1次,共7次,停药恢复28 d。应用流式细胞仪进行外周血中淋巴细胞分群检测,检测经TC007刺激后分泌干扰素-γ（IFN-γ）的特异T细胞水平;免疫组化检测肝脏IFN-γ、肿瘤坏死因子-α（TNF-α）、白细胞介素-2（IL-2）表达;试剂盒法测定血清中抗Ad5抗体的中和活性,补体成分C3、C4及免疫复合物（CIC）水平;给药期及恢复期结束,进行多个组织取材,HE染色后进行病理学阅片,TaqMan探针qPCR法进行Ad5分布检测。结果 与溶媒对照组比较,各组动物血浆中CD3⁺CD8⁺T细胞、CD4⁺/CD8⁺、CD3^-CD56⁺NK细胞、CD3⁺CD56⁺NKT细胞、巨噬细胞CD45⁺CD14⁺百分率均未见明显异常;Ad5-Null组特异T细胞均未见明显改变,TC007各剂量组针对HBV 3种抗原（Core、Env、Pol）分泌IFN-γ的特异性T细胞水平显著增加（P<0.05、0.01）,停药恢复末期（d68）,效应值仍略高;TC007各剂量组和Ad5-Null组食蟹猴肝脏组织中IFN-γ、TNF-α、IL-2均高于溶媒对照组,且呈剂量相关性,但TC007各剂量组与Ad5-Null组比较未见明显差异;TC007各剂量、Ad5-Null组动物均产生抗Ad5抗体,39/40例具有中和活性;TC007各剂量组补体C3、C4未见明显异常,高剂量组CIC轻微升高（P<0.05）。Ad5主要分布于给药局部,在腋下淋巴结有少量分布,其他组织未见分布。TC007及Ad5-Null组均可见给药局部轻微至轻度皮下组织炎性细胞浸润,停药恢复28 d,病变可见明显恢复,其他组织未见病理学改变。结论 TC007在食蟹猴体内具有一定免疫原性,抗Ad5抗体具有中和活性,高剂量组可见轻微免疫复合物形成,未见免疫器官损伤,生物分布主要集中于给药部位,可产生特异性T细胞。     

Preclinical safety evaluation of levonadifloxacin,a novel anti-methicillin-resistant Staphyloccocus aureus benzoquinolizine fluoroquinolone by intravenous and oral administration

Fluoroquinolone (FQ) antibacterials have drawn heightened attention from various international regulatory agencies due to their class-specific side effects. Levonadifloxacin is a novel broad spectrum benzoquinolizine FQ active against methicillin-resistant Staphyloccocus aureus (MRSA). Owing to FQ-associated safety concerns, extensive preclinical safety pharmacology (central nervous system and cardiac safety) and toxicology studies (subacute repeat-dose toxicity, genotoxicity, phototoxicity and chondrotoxicity) of levonadifloxacin were performed at relatively high doses. Intravenous (IV) and oral studies were conducted using WCK 771 (l -arginine salt of levonadifloxacin) and WCK 2349 (l -alanine ester prodrug of levonadifloxacin), respectively. Safety pharmacology studies following single dose revealed no adverse effects on central nervous system (including seizure) in mice and cardiovascular system (hERG and monkey telemetry). In repeat-dose toxicity studies, except for IV bolus dosing related effects in rat (hyperactivity, mild convulsion, polypnoea and injection site irritation) and dog (emesis and salivation), no other adverse findings limiting the dosing duration were observed. No major biochemical, haematological, gross or histopathological changes suggestive of damage to vital organs were observed in either WCK 771- or WCK 2349-treated groups. WCK 771 and WCK 2349 were found to be nongenotoxic; however, they showed weak phototoxicity that was comparable with levofloxacin. WCK 771 showed chondrotoxicity in the Beagle dog pups on repeat-dose administration; however, the severity level was lower than ofloxacin. Overall, preclinical safety studies helped establish wider safety margin for WCK 771 and WCK 2349 that supports administration of higher therapeutic doses in humans by both IV and oral routes, thereby enabling safe anti-MRSA treatment.     

Nonclinical strategy considerations for safety pharmacology: evaluation of biopharmaceuticals

Introduction: Biopharmaceuticals, such as monoclonal antibodies and recombinant peptides, are important therapeutics to treat human disease. Key features of biopharmaceuticals that make them innovative medicines are their clinical effectiveness, high specificity for their human target, long half-life and target coverage, and low risk for “off-target” pharmacology.

Areas covered: This paper describes nonclinical safety pharmacology assessment of biopharmaceuticals with an emphasis on special considerations needed for these agents. Insight is provided on various approaches to conduct safety pharmacology studies for such therapeutics, including appropriate integration into non-rodent toxicity studies.

Expert opinion: The safety pharmacology evaluation of biopharmaceuticals requires a science-based, case-by-case approach, as each biological modality will have unique pharmacological characteristics that influence the overall nonclinical safety assessment strategy. The integration of safety pharmacology endpoints into general (repeat-dose) toxicity studies is a rational paradigm for assessing potential changes in the cardiovascular, central nervous, and respiratory systems, but requires thoughtful and practical planning. In some cases, especially based on target-pharmacology concerns, dedicated and optimally designed safety pharmacology studies may be needed to assess the functional risk of a new biopharmaceutical. For example, cardiovascular telemetry studies may be needed to detect small changes in arterial blood pressure after acute and chronic exposure.     

Preclinical safety evaluation of recombinant human interleukin-10

Escherichia coli-derived recombinant human interleukin-10 (rhuIL-10) has been evaluated in an extensive series of in vivo and in vitro nonclinical safety studies, including genetic toxicology, single- and repeat-dose systemic toxicity and toxicokinetics, reproductive toxicity, and specialized irritation studies. The primary test species in the toxicology studies were the mouse and monkey based on rhuIL-10 activity in receptor binding and ex vivo cytokine assays. Supported by a detailed preclinical program of therapeutic and prophylactic animal models in autoimmune diseases, the initial clinical development program has focused on investigating the therapeutic potential of rhuIL-10 (Tenovil) in Crohn's disease and rheumatoid arthritis. The results of the subcutaneous toxicity studies, up to 3 months dosing duration in mice and 6 months dosing duration in monkeys, support the development of rhuIL-10 for present and future clinical indications by the subcutaneous route of administration.     

Antisense oligonucleotides on neurobehavior,respiratory, and cardiovascular function,and hERG channel current studies

IntroductionSafety Pharmacology studies were conducted in mouse, rat, and non-human primate to determine in vivo effects of antisense oligonucleotides (ASOs) on the central nervous system, respiratory system, and cardiovascular system. Effects on the hERG potassium channel current was evaluated in vitro.MethodsASOs contained terminal 2′-O-methoxyethyl nucleotides, central deoxy nucleotides, and a phosphorothioate backbone. Neurobehavior was evaluated by Functional Observatory Battery in rodents. Respiratory function was directly measured in rodents by plethysmograph; respiratory rate and blood gases were measured in monkey. Basic cardiovascular endpoints were measured in rat; cardiovascular evaluation in monkey involved implanted telemetry units. In single and repeat dose studies ASOs were administered by subcutaneous injection at up to 300 mg/kg, 250 mg/kg, and 40 mg/kg in mouse, rat, or monkey, respectively. Assays were performed in HEK293 or CHO-K1 cells, stably transfected with hERG cDNA, at ASO concentrations of up to 300 μM.ResultsNo apparent effects were noted for respiratory or CNS function. Continuous monitoring of the cardiovascular system in monkey demonstrated no ASO-related changes in blood pressures, heart rate, or ECG and associated parameters (i.e., QRS duration). Specific assessment of the hERG potassium channel indicated no potential for actions on ventricular repolarization or modest effects only at excessive concentrations.DiscussionThe absence of direct actions on neurobehavior and respiratory function associated with the administration of ASOs in safety pharmacology core battery studies is consistent with published toxicology studies. The combination of in vitro hERG studies and in vivo studies in rat and monkey are consistent with no direct actions by ASOs on cardiac cell function or electrical conduction at relevant concentrations and dose levels. Taken as a whole, dedicated studies focused on the safety pharmacology of specific organ systems do not appear to add significant data for interpretation of potential adverse effects. The need for dedicated studies for future ASOs in the same class is questionable, as a more encompassing data set can be collected in repeat dose and longer-term toxicology studies.     

Pre-clinical toxicity and immunogenicity evaluation of a MUC1–MBP/BCG anti-tumor vaccine

Mucin 1 (MUC1), as an oncogene, plays a key role in the progression and tumorigenesis of many human adenocarcinomas and is an attractive target in tumor immunotherapy. Our previous study showed that the MUC1–MBP/BCG anti-tumor vaccine induced a MUC1-specific Th1-dominant immune response, simulated MUC1-specific cytotoxic T lymphocyte killing activity, and could significantly inhibit MUC1-expression B16 cells' growth in mice. To help move the vaccine into a Phase I clinical trial, in the current study, a pre-clinical toxicity and immunogenicity evaluation of the vaccine was conducted. The evaluation was comprised of a single-dose acute toxicity study in mice, repeat-dose chronic toxicity and immunogenicity studies in rats, and pilot toxicity and immunogenicity studies in cynomolgus monkeys. The results showed that treatment with the MUC1–MBP/BCG anti-tumor vaccine did not cause any organ toxicity, except for arthritis or local nodules induced by BCG in several rats. Furthermore, the vaccine significantly increased the levels of IFN-γ in rats, indicating that Th1 cells were activated. In addition, the results showed that the MUC1–MBP/BCG anti-tumor vaccine induced a MUC1-specific IgG antibody response both in rats and cynomolgus monkeys. Collectively, these data are beneficial to move the MUC1–MBP/BCG anti-tumor vaccine into a Phase I clinical trial.     

含硼小分子固醇调控元件结合蛋白抑制剂的合成及其体外抗肿瘤活性研究

目的设计、合成固醇调控元件结合蛋白(SREBP)抑制剂BF-175的衍生物,并对其稳定性和体外抗肿瘤活性进行研究。方法以水杨醛为起始原料,通过6步反应合成了一系列含硼小分子SREBP抑制剂,通过高效液相色谱法分析其稳定性,并采用RT-PCR实验评价其抗肿瘤活性。结果合成了9个含硼小分子SREBP抑制剂,并通过NMR和MS对其进行表征。其药学稳定性均优于BF-175。RT-PCR检测表明目标化合物可以抑制人子宫内膜腺癌细胞(AN3CA)中SREBP靶基因硬脂酰辅酶A去饱和酶-1(SCD-1)的表达,其中化合物7g的活性高于BF-175。结论含硼小分子SREBP抑制剂具有较好的抗肿瘤活性,为此类药物的作用机制研究奠定了基础。     

QT PRODACT: in vivo QT assay with a conscious monkey for assessment of the potential for drug-induced QT interval prolongation

In safety pharmacology studies, the effects on the QT interval of electrocardiograms are routinely assessed using a telemetry system in cynomolgus monkeys. However, there is a lack of integrated databases concerning in vivo QT assays in conscious monkeys. As part of QT Interval Prolongation: Project for Database Construction (QT PRODACT), the present study examined 10 positive compounds with the potential to prolong the QT interval and 6 negative compounds considered to have no such effect on humans. The experiments were conducted at 7 facilities in accordance with a standard protocol established by QT PRODACT. The vehicle or 3 doses of each test compound were administered orally to male cynomolgus monkeys (n=3-4), and telemetry signals were recorded for 24 h. None of the negative compounds prolonged the corrected QT using Bazett's formula (QTcB) interval. On the other hand, almost all of the positive compounds prolonged the QTcB interval, but haloperidol, terfenadine, and thioridazine did not. The failure to detect the QTcB interval prolongation appeared to be attributable for the differences in metabolism between species and/or disagreement with Bazett's formula for tachycardia. In the cynomolgus monkeys, astemizole induced Torsade de Pointes and cisapride caused tachyarrhythmia at lower plasma concentrations than those observed in humans and dogs. These results suggest that in vivo QT assays in conscious monkeys represent a useful model for assessing the risks of drug-induced QT interval prolongation.     

Preclinical development of AMG 139, a human antibody specifically targeting IL-23

BACKGROUND AND PURPOSE

AMG 139 is a human anti-IL-23 antibody currently in a phase II trial for treating Crohn''s disease. To support its clinical development in humans, in vitro assays and in vivo studies were conducted in cynomolgus monkeys to determine the pharmacology, preclinical characteristics and safety of this monoclonal antibody.

EXPERIMENTAL APPROACH

The in vitro pharmacology, pharmacokinetics (PK), pharmacodynamics and toxicology of AMG 139, after single or weekly i.v. or s.c. administration for up to 26 weeks, were evaluated in cynomolgus monkeys.

KEY RESULTS

AMG 139 bound with high affinity to both human and cynomolgus monkey IL-23 and specifically neutralized the biological activity of IL-23 without binding or blocking IL-12. After a single dose, linear PK with s.c. bioavailability of 81% and mean half-life of 8.4–13 days were observed. After weekly s.c. dosing for 3 or 6 months, AMG 139 exposure increased approximately dose-proportionally from 30 to 300 mg·kg⁻¹ and mean accumulation between the first and last dose ranged from 2- to 3.5-fold. Peripheral blood immunophenotyping, T-cell-dependent antigen responses and bone formation markers were not different between AMG 139 and vehicle treatment. No adverse clinical signs, effects on body weight, vital signs, ophthalmic parameters, clinical pathology, ECG, organ weights or histopathology were observed in the monkeys with the highest dose of AMG 139 tested (300 mg·kg⁻¹ s.c. or i.v.).

CONCLUSIONS AND IMPLICATIONS

The in vitro pharmacology, PK, immunogenicity and safety characteristics of AMG 139 in cynomolgus monkeys support its continued clinical development for the treatment of various inflammatory diseases.