固相萃取离子色谱法测定反义寡核苷酸药物杂质

目的建立同时测定寡核苷酸原料药中杂质醋酸（MCAA）、二氯乙酸（DCAA）、氯离子（Cl－）含量的固相萃取离子色谱（IC）法。方法Ionpac AS 23（4 mm×250 mm）分析柱;Ionpac AG 23（4 mm×50 mm）保护柱;流动相：0.8 mmol•L－1 碳酸氢钠/4.5 mmol•L－1碳酸钠;抑制型电导检测器。结果MCAA、DCAA和Cl－的标准曲线方程分别为：Y＝ 5.230 7X + 0.014 （r＝0.999 7）,Y＝ 19.762X－0.007 2 （r＝0.999 9）和Y＝507.4X－0.746 2 （r＝0.999 8）;MCAA在0.002～0.400 μg•mL－1浓度范围内呈良好的线性关系,DCAA在0.000 624～0.156 μg•mL－1浓度范围内呈良好的线性关系,Cl－在2.4×10－3～1.5 mg•mL－1浓度范围内呈良好的线性关系,其检测限分别为0.02,0.312 ng•mL－1和0.01 μg•mL－1。结论该方法操作简便、快速,结果准确,可用于反义寡核苷酸原料药中杂质的控制。     

靶向微小RNA的病毒感染疾病治疗新策略

微小RNA(microRNA,miRNA)是真核生物细胞的一类长度约为19～25个核苷酸的非编码小RNA,在生物体发育和基因表达过程发挥重要的调控作用。miRNA分子通过与靶mRNA的互补配对,在转录后水平上对基因的表达进行负调控。近来研究表明miRNA在病毒感染宿主过程中发挥重要作用。通过设计抗miRNA反义寡核苷酸促进或阻断特定miRNA的表达可以实现抑制病毒复制的目标。本文综述了抗miRNA反义寡核苷酸研究现状和面临的挑战。     

VEGF反义寡核苷酸与顺铂联合治疗抑制裸鼠骨肉瘤的生长

目的探讨血管内皮生长因子（VEGF）反义寡核苷酸和顺铂单用或联用对裸鼠骨肉瘤生长的抑制作用。方法制备BALC／C裸鼠皮下骨肉瘤模型24只,随机分为4组：VEGF反义寡核苷酸治疗（ASODN）组、顺铂治疗（DDP）组、联合治疗（ASODN＋DDP）、组和对照组,每组6只。接种MG-63人成骨肉瘤细胞后24h内采用瘤内注射和腹腔给药的方式,以VEGF反义寡核苷酸或（和）顺铂进行治疗;观察各组稞鼠肿瘤的生长情况、裸鼠的一般情况与生存期。断颈处死裸鼠,测量肿瘤体积大小,称量肿瘤的重量。结果VEGF—ASODN治疗组、DDP治疗组和联合治疗组裸鼠骨肉瘤的生长均受到不同程度抑制,抑瘤率分别为44．56％、53．69％和86．80％,以联合治疗组的疗效最佳。结论VEGF反义寡核苷酸与顺铂联用比单用顺铂可更好地抑制骨肉瘤的生长,两者联用可发挥协同抗瘤效应。     

Quantum rods as nanocarriers of gene therapy

Both antisense oligonucleotides (ASODN) and small interfering RNA (siRNA) have enormous potential to selectively silence specific cancer-related genes and could therefore be developed to be important therapeutic anti-cancer drugs. The use of nanotechnology may allow for significant advancement of the therapeutic potential of ASODN and siRNA, due to improved pharmacokinetics, bio-distribution and tissue specific targeted therapy. In this mini-review, we have discussed the advantages of using a nanocarrier such as a multimodal quantum rod (QR) complexed with siRNA for gene delivery. Comparisons are made between ASODN and siRNA therapeutic efficacies in the context of cancer and the enormous application potential of nanotechnology in oncotherapy is discussed. We have shown that a QR-interleukin-8 (IL-8) siRNA nanoplex can effectively silence IL-8 gene expression in the PC-3 prostate cancer cells with no significant toxicity. Thus, nanocarriers such as QRs can help translate the potent effects of ASODN/siRNA into a clinically viable anti-cancer therapy. Drug delivery for cancer therapy, with the aid of nanotechnology is one of the major translational aspects of nanomedicine, and efficient delivery of chemotherapy drugs and gene therapy drugs or their co-delivery continue to be a major focus of nanomedicine research.     

Targeting endogenous inhibitors of apoptosis for treatment of cancer,stroke and multiple sclerosis

The inhibitor of apoptosis (IAP) genes have emerged as probably the most important intrinsic regulators of apoptosis. The members of the IAP family are highly conserved in evolutionarily distant species and perform the critical role of binding to and inhibiting distinct caspases. This inhibition is mediated by discrete baculoviral IAP repeat domains that, in a domain-specific manner, inhibit either the initiator or executioner caspases. As such the function of IAPs lies at the very centre of virtually all apoptotic pathways. Since many, if not most, human pathologies involve aberrant apoptosis, the modulation of IAP levels or their activity offers huge therapeutic potential for treatment of various disorders. Indeed, available data suggest that the therapeutic downregulation of IAPs by antisense targeting or their adenovirally-mediated overexpression, can in fact be used to successfully modulate cell death.     

磷脂酰肌醇3-激酶调控白介素-18诱导核因子-кB活化(英文)

目的:研究磷脂酰肌醇(PI)3-激酶在白介素18(IL-18)诱导核因子-кB(NF-кB)活化中的作用。方法:Lipofectin介导反义PI 3-激酶寡核苷酸转染HepG2细胞。用逆转录PCR法检测PI 3-激酶mRNA表达水平,以Sandwich ELISA法检测NF-кB的活化。结果:(1)反义PI 3-激酶寡核苷酸抑制PI 3-激酶mRNA表达。(2)IL-18诱导NF-кB活化。(3)反义PI3-激酶寡核苷酸呈时间(5-24h)和浓度(1-8mg/L)依赖性地抑制IL-18诱导的NF-кB活化。结论:PI 3-激酶调控白介素-18诱导的NF-кB活化。     

Organ distribution and stability of phosphorothioated oligodeoxyribonucleotides in mice.

Results of recent studies in our laboratory have suggested a potential role for antisense oligodeoxyribonucleotides (oligo(dN)s) as therapeutic agents in the treatment of human hepatitis B virus infection. As a first step towards assessing the potential utility of oligo(dN) in therapy, we have examined the organ distribution, stability and toxicity of a phosphorothioated oligo(dN) (S-oligo) of 20 nucleotides in length which was administered to mice via different routes. Among the various organs analysed, the liver retained the highest amount of S-oligo (1.3-2 per cent of the total injection) at the peak time (10-30 min) regardless of the route of injection. However, the S-oligo appeared to be degraded in the liver to about 40 per cent of its original length within 30 min of injection, presumably by the action of 3' exonucleases. Injection of doses of up to 5 mg kg-1 of S-oligo had no apparent toxic effects on the mice.     

Therapeutic Ultrasound for Gene Delivery

In vitro and in vivo studies using perfluorocarbon-exposed sonicated dextrose albumin (PESDA) microbubbles to enhance gene delivery are reviewed. In vitro studies show PESDA binds to oligonucleotides and that ultrasound can be used to deposit these nucleotides. In addition, in vitro studies show that drug release from microspheres is dependent on ultrasound transmission frequency as well as pulsed or continuous application. Early in vivo studies confirm that ultrasound in combination with microbubbles can be used to facilitate gene deposition. However, the role of ultrasound targeting gene delivery remains to be determined.