Supramolecular assembly of Toll-like receptor 7/8 agonist into multimeric water-soluble constructs enables superior immune stimulation in vitro and in vivo.
Supramolecular assembly of Toll-like receptor 7/8 agonist into multimeric water-soluble constructs enables superior immune stimulation in vitro and in vivo.
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DOI：
10.1021/acsabm.0c00189
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发表时间：
2020-05-18
影响因子：
4.7
通讯作者：
Fuerst TR
中科院分区：
文献类型：
--
作者：
Andrianov AK;Marin A;Wang R;Karauzum H;Chowdhury A;Agnihotri P;Yunus AS;Mariuzza RA;Fuerst TR
Resiquimod 或 R848 (RSQD) 是一种 Toll 样受体 (TLR) 7/8 激动剂，由于其促进高度理想的细胞免疫的潜力，有望成为疫苗佐剂。迄今为止，这种小分子在该领域的发展在很大程度上受到其体内快速清除和缺乏与疫苗抗原结合的阻碍。在这里，我们报告了纳米级尺寸的多聚体 TLR 7/8 构建体，该构建体是 RSQD 与水溶性临床阶段聚合物 - 聚[二（羧基苯氧基）磷腈]（PCPP）自发自组装的结果。离子配对构建体 (PCPP-R) 和三元复合物（还包括丙型肝炎病毒 (HCV) 抗原）的形成已通过动态光散射 (DLS)、比浊法、荧光光谱、不对称流场流分级 (AF4) 和 1H NMR 光谱方法得到证明。由此产生的超分子组装体 PCPP-R 在细胞测定（小鼠巨噬细胞报告细胞系）中实现了卓越的免疫刺激，并显示出改善的体外血液相容性（人红细胞）。体内研究表明，PCPP-R 佐剂 HCV 制剂在 BALB/c 小鼠中诱导更高的血清中和滴度，并将反应转向理想的细胞免疫，通过抗体同种型比率 (IgG2a/IgG1) 和细胞因子分泌脾细胞的离体分析（更高水平的干扰素 γ (IFN-γ) 单和肿瘤坏死因子 α (TNF-α)/IFN-γ 双产生细胞）进行评估。非共价多聚化方法与之前提出的涉及共价缀合或封装的 RSQD 递送方法形成鲜明对比，并提供了一种灵活的方法，可以与其他肠胃外给药药物整合。
Resiquimod or R848 (RSQD) is a Toll-like receptor (TLR) 7/8 agonist which shows promise as vaccine adjuvant due to its potential to promote highly desirable cellular immunity. The development of this small molecule in the field to date has been largely impeded by its rapid in vivo clearance and lack of association with vaccine antigens. Here, we report a multimeric TLR 7/8 construct of nano-scale size, which results from a spontaneous self-assembly of RSQD with a water-soluble clinical-stage polymer - poly[di(carboxylatophenoxy)phosphazene] (PCPP). The formation of ionically paired construct (PCPP-R) and a ternary complex, which also includes Hepatitis C virus (HCV) antigen, has been demonstrated by dynamic lights scattering (DLS), turbidimetry, fluorescence spectroscopy, asymmetric flow field flow fractionation (AF4), and 1H NMR spectroscopy methods. The resulting supramolecular assembly PCPP-R enabled superior immunostimulation in cellular assays (mouse macrophage reporter cell line) and displayed improved in vitro hemocompatibility (human erythrocytes). In vivo studies demonstrated that PCPP-R adjuvanted HCV formulation induced higher serum neutralization titers in BALB/c mice and shifted the response towards desirable cellular immunity, as evaluated by antibody isotype ratio (IgG2a/IgG1) and ex vivo analysis of cytokine secreting splenocytes (higher levels of interferon gamma (IFN-γ) single and tumor necrosis factor alpha (TNF-α)/IFN-γ double producing cells). The non-covalent multimerization approach stands in contrast to previously suggested RSQD delivery methods, which involve covalent conjugation or encapsulation, and offers a flexible methodology that can be potentially integrated with other parenterally administered drugs.
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影响因子：
4
作者：
Andrianov AK;Marin A;Fuerst TR
通讯作者：
Fuerst TR
影响因子：
6.2
作者：
Andrianov, AK;Marin, A;Chen, JP
通讯作者：
Chen, JP
影响因子：
6.2
作者：
Fauvelle C;Colpitts CC;Keck ZY;Pierce BG;Foung SK;Baumert TF
通讯作者：
Baumert TF
影响因子：
6.2
作者：
Andrianov, Alexander K.;Marin, Alexander;Fuerst, Thomas R.
通讯作者：
Fuerst, Thomas R.
DOI：
10.1002/chem.201702942
发表时间：
2017-12-14
期刊：
Chemistry (Weinheim an der Bergstrasse, Germany)
影响因子：
--
作者：
Aichhorn S;Linhardt A;Halfmann A;Nadlinger M;Kirchberger S;Stadler M;Dillinger B;Distel M;Dohnal A;Teasdale I;Schöfberger W
通讯作者：
Schöfberger W