Retargeted clostridial endopeptidases: Inhibition of nociceptive neurotransmitter release in vitro, and antinociceptive activity in in vivo models of pain
Retargeted clostridial endopeptidases: Inhibition of nociceptive neurotransmitter release in vitro, and antinociceptive activity in in vivo models of pain
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DOI：
10.1002/mds.20008
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发表时间：
2004-03-01
影响因子：
8.6
通讯作者：
Foster, KA
中科院分区：
文献类型：
--
作者：
Chaddock, JA;Purkiss, JR;Foster, KA
梭菌神经毒素有效且特异性地抑制特定细胞类型中神经递质的释放。先前报道的数据表明，A 型肉毒杆菌神经毒素的催化活性 LH(N) 内肽酶片段（称为 LH(N)/A）可以在体外重新靶向一系列细胞类型，从而抑制一系列递质的分泌。在这里，我们报道了与脊髓神经元相比，肽链内切酶缀合物的合成，其对伤害性传入神经具有体外选择性。在体外和体内疼痛模型中评估了刺桐凝集素和 LH(N)/A 之间制备的化学缀合物。对 E.cristagalli 凝集素与天然来源的 LH(N)/A 或从大肠杆菌纯化的重组 LH(N)/A 之间制备的化学缀合物进行了评估，并证明了重组材料的等效性。还评估了缀合物在体外抑制神经递质释放的作用持续时间，并且与用肉毒梭菌神经毒素观察到的作用持续时间相当。靶向选择性和治疗潜力已通过体内电生理学研究得到证实。此外，已在体内疼痛模型中评估了缀合物的镇痛特性并观察到延长持续时间的效果。这些数据为重定向梭菌内肽酶作为新型镇痛药的概念提供了原理证明。 (C) 2004 年运动障碍协会。
Clostridial neurotoxins potently and specifically inhibit neurotransmitter release in defined cell types. Previously reported data have demonstrated that the catalytically active LH(N) endopeptidase fragment of botulinum neurotoxin type A (termed LH(N)/A) can be retargeted to a range of cell types in vitro to lead to inhibition of secretion of a range of transmitters. Here, we report the synthesis of endopeptidase conjugates with in vitro selectivity for nociceptive afferents compared to spinal neurons. Chemical conjugates prepared between Erythrina cristagalli lectin and LH(N)/A are assessed in vitro and in in vivo models of pain. Chemical conjugates prepared between E. cristagalli lectin and either natively sourced LH(N)/A, or recombinant LH(N)/A purified from Escherichia coli are assessed, and equivalence of the recombinant material is demonstrated. The duration of action of inhibition of neurotransmitter release by the conjugate in vitro is also assessed and is comparable to that observed with Clostridium botulinum neurotoxin. Selectivity of targeting and therapeutic potential have been confirmed by in vivo electrophysiology studies. Furthermore, the analgesic properties of the conjugate have been assessed in in vivo models of pain and extended duration effects observed. These data provide proof of principle for the concept of retargeted clostridial endopeptidases as novel analgesics. (C) 2004 Movement Disorder Society.