Sewell K Lea, Geary Richard S, Baker Brenda F, Glover Josephine M, Mant Timothy G K, Yu Rosie Z, Tami Joseph A, Dorr F Andrew
Isis Pharmaceuticals Inc., Carlsbad, California 92008-7208, USA.
J Pharmacol Exp Ther. 2002 Dec;303(3):1334-43. doi: 10.1124/jpet.102.036749.
ISIS 104838 is a 20-mer phosphorothioate antisense oligonucleotide (ASO) that binds tumor necrosis factor-alpha (TNF-alpha) mRNA. It carries a 2'-methoxyethyl modification on the five 3' and 5' nucleotide sugars, with 10 central unmodified deoxynucleotides. ISIS 104838 was identified from a 264 ASO screen in phorbol myristate acetate-activated keratinocytes, and the dose response was assessed in lipopolysaccharide (LPS)-activated monocytes. Healthy males received multiple intravenous (i.v.) ISIS 104838 infusions in a placebo-controlled dose escalation trial (0.1-6 mg/kg). Additional volunteers received single or multiple subcutaneous (s.c.) injections. ISIS 104838 suppressed TNF-alpha protein by 85% in stimulated keratinocytes. The IC50 for TNF-alpha mRNA inhibition in stimulated monocytes was <1 microM. For i.v., C(max) occurred at the end of infusion. The effective plasma half-life was 15 to 45 min at 0.1 to 0.5 mg/kg and 1 to 1.8 h for higher doses. The apparent terminal plasma elimination half-life approximated 25 days. Obese subjects had higher plasma levels following equivalent mg/kg doses. For s.c. injections, C(max) occurred at 2 to 4 h and was lower than with equivalent i.v. dosing. Plasma bioavailability compared with i.v. was 82% following a 200 mg/ml s.c. injection. Transient activated partial thromboplastin time prolongation occurred after i.v. infusions and minimally after s.c. injections. Two subjects experienced rash, one a reversible platelet decrease, and mild injection site tenderness was noted. TNF-alpha production by peripheral blood leukocytes, induced ex vivo by LPS, was decreased by ISIS 104838 (p < 0.01). ISIS 104838, a second-generation antisense oligonucleotide, was generally well tolerated intravenously and subcutaneously. The pharmacokinetics support an infrequent dosing interval. Inhibition of TNF-alpha production ex vivo was demonstrated.
ISIS 104838是一种20聚体硫代磷酸酯反义寡核苷酸（ASO），可与肿瘤坏死因子-α（TNF-α）信使核糖核酸（mRNA）结合。它在5个3'和5'核苷酸糖上带有2'-甲氧基乙基修饰，中间有10个未修饰的脱氧核苷酸。ISIS 104838是从佛波酯肉豆蔻酸酯乙酸酯激活的角质形成细胞中的264种ASO筛选中鉴定出来的，并在脂多糖（LPS）激活的单核细胞中评估了剂量反应。健康男性在一项安慰剂对照的剂量递增试验（0.1 - 6毫克/千克）中接受多次静脉注射ISIS 104838。其他志愿者接受单次或多次皮下注射。ISIS 104838在受刺激的角质形成细胞中可将TNF-α蛋白抑制85%。在受刺激的单核细胞中，抑制TNF-α mRNA的半数抑制浓度（IC50）<1微摩尔。静脉注射时，最大血药浓度（C(max)）在输注结束时出现。在0.1至0.5毫克/千克剂量下，有效血浆半衰期为15至45分钟，更高剂量时为1至1.8小时。表观终末血浆消除半衰期约为25天。肥胖受试者在给予等效毫克/千克剂量后血浆水平较高。皮下注射时，C(max)在2至4小时出现，且低于等效静脉注射剂量。皮下注射200毫克/毫升后，与静脉注射相比，血浆生物利用度为82%。静脉输注后出现短暂的活化部分凝血活酶时间延长，皮下注射后则轻微延长。两名受试者出现皮疹，一名受试者血小板可逆性减少，并观察到轻微的注射部位压痛。ISIS 104838可降低LPS体外诱导的外周血白细胞产生的TNF-α（p < 0.01）。第二代反义寡核苷酸ISIS 104838静脉内和皮下给药时一般耐受性良好。药代动力学支持不频繁的给药间隔。已证明其在体外可抑制TNF-α的产生。
