A New Method to Produce MonoPEGylated Dimeric Cytokines Shown with Human Interferon-α2b
A New Method to Produce MonoPEGylated Dimeric Cytokines Shown with Human Interferon-α2b
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DOI：
10.1021/bc9001773
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发表时间：
2009-10-01
影响因子：
4.7
通讯作者：
Goldenberg, David M.
中科院分区：
文献类型：
--
作者：
Chang, Chien-Hsing;Rossi, Edmund A.;Goldenberg, David M.
我们已经将时钟锁定(DNL)方法改进为一种新的聚乙二醇化技术，以人干扰素-α2b(干扰素-α2b)为例。DNL的中心是一对不同的蛋白质结构域，参与cAMP依赖的蛋白激酶(PKA)和A-激酶锚定蛋白(AKAP)之间的自然联系。这些结构域是聚乙二醇二聚体形式的干扰素-α2b与聚乙二醇特异性结合的连接物。包含干扰素-α2b的融合蛋白和PKA的二聚化和对接结构域(DDD)与相互作用的AKAP的聚乙二醇化锚定结构域(AD)相结合，可以很容易地形成包含两个拷贝的干扰素-a2b和单个位点特异性连接的聚乙二醇链的三聚体复合体。已经产生了三个这样的干扰素-a2b的单聚乙二醇化二聚体，第一个具有20 kDa的线性聚乙二醇化，称为α2b-362，第二个具有30 kDa的线性聚乙二醇化(α2b-413)，以及第三个具有40 kDa支化的聚乙二醇化的干扰素-a2b(α2b-457)。这三种药物在体外都保持了抗病毒和抗肿瘤的活性，并在小鼠身上表现出改善的药代动力学特性，这在Daudi人淋巴瘤异种移植模型中转化为有效和持久的治疗效果。我们预计DNL方法在开发二聚体和单聚乙二醇化的长效疗法方面具有广泛的适用性，生物利用度增加，允许较少的频繁给药。
We have adapted the clock-and-lock (DNL) method, into a novel PEGylation technology using human interferon-alpha 2b (IFN-alpha 2b) as an example. Central to DNL is a pair of distinct protein domains involved in the natural association between cAMP-dependent protein kinase (PKA) and A-kinase anchoring proteins (AKAPs). These domains serve as linkers for site-specific conjugation of poly(ethylene glycol) (PEG) to a dimeric form of IFN-alpha 2b. The combination of a fusion protein comprising IFN-alpha 2b and the dimerization-and-docking domain (DDD) of PKA with a PEG-derivatized anchoring domain (AD) of an interactive AKAP results in facile formation of a trimeric complex containing two copies of IFN-a2b and a single site-specifically linked PEG chain. Three such monoPEGylated dimers of IFN-a2b have been generated, the first with a 20 kDa linear PEG, referred to as alpha 2b-362, the second with a 30 kDa linear PEG (alpha 2b-413), and the third with a 40 kDa branched PEG (alpha 2b-457). All three retained antiviral and antitumor activity in vitro and showed improved pharmacokinetic properties in mice, which translated into potent and prolonged therapeutic efficacy in the Daudi human lymphoma xenograft model. We anticipate wide applicability of the DNL method for developing long-acting therapeutics that are dimeric and monoPEGylated with the increased bioavialability allowing for less frequent dosing.