Chemoenzymatic synthesis of cryptophycin anticancer agents by an ester bond-forming non-ribosomal peptide synthetase module.
Chemoenzymatic synthesis of cryptophycin anticancer agents by an ester bond-forming non-ribosomal peptide synthetase module.
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DOI：
10.1021/ja204716f
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发表时间：
2011-09-21
影响因子：
15
通讯作者：
Sherman, David H.
中科院分区：
文献类型：
--
作者：
Ding, Yousong;Rath, Christopher M.;Bolduc, Kyle L.;Hakansson, Kristina;Sherman, David H.
隐藻素是一组具有抗耐药肿瘤活性的蓝藻缩肽。虽然被证明是有前途的，但需要进一步的努力，通过具有改进的药用特性的新一代类似物将这些高效化合物返回临床。在此，我们报告了依赖于多功能酶CrpD-M2的化学合成路线，该酶将2-羟基酸部分（单元D）并入念珠藻素类似物中。CrpD-M2是一种独特的非核糖体肽合成酶（NRPS）模块，由缩合-腺苷酸化-酮还原-硫醇化（C-A-KR-T）结构域组成。我们询问了A-结构域2-酮和2-羟基酸活化和加载，以及在NADPH和NADH存在下的KR结构域活性。通过CrpD-M2 C结构域催化的酯键形成，用三种合成的念珠藻素链延长中间体类似物（SNAC-ABC）延长所得的2-羟基酸。最后，酶结合的裂念珠藻素产物被CRP硫酯酶（TE）大环内酯化。通过酶结合中间体和产物的液相色谱傅里叶变换离子回旋共振质谱（LC-FTICR-MS）分析，能够对这些连续步骤进行分析。这种新的化学酶促合成念珠藻素涉及四个连续的催化步骤，导致在最终的链延长中间体的2-羟基酸部分的掺入。这是第一个例子，其中NRPS嵌入KR结构域用于组装一个完全精心制作的天然产物，并作为一个证明的原则，化学酶合成新的念珠藻素类似物。
Cryptophycins are a group of cyanobacterial depsipeptides with activity against drug-resistant tumors. Although shown to be promising, further efforts are required to return these highly potent compounds to the clinic through a new generation of analogs with improved medicinal properties. Herein, we report a chemosynthetic route relying on the multifunctional enzyme CrpD-M2 that incorporates a 2-hydroxy-acid moiety (unit D) into cryptophycin analogs. CrpD-M2 is a unique non-ribosomal peptide synthetase (NRPS) module comprised of condensation-adenylation-ketoreduction-thiolation (C-A-KR-T) domains. We interrogated A-domain 2-keto and 2-hydroxy acid activation and loading, and KR domain activity in the presence of NADPH and NADH. The resulting 2-hydroxy acid was elongated with three synthetic cryptophycin chain elongation intermediate analogs (SNAC-ABC) through ester bond formation catalyzed by CrpD-M2 C domain. Finally, the enzyme bound seco-cryptophycin products were macrolactonized by the Crp thioesterase (TE). The analysis of these sequential steps was enabled through liquid chromatography Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS) analysis of enzyme bound intermediates and products. This novel chemoenzymatic synthesis of cryptophycin involves four sequential catalytic steps leading to the incorporation of a 2-hydroxy acid moiety in the final chain elongation intermediate. This is the first example where a NRPS-embedded KR domain is employed for assembly of a fully elaborated natural product, and serves as a proof-of-principle for chemoenzymatic synthesis of new cryptophycin analogs.
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影响因子：
16.6
作者：
Seufert, Wolfgang;Beck, Zachary Q.;Sherman, David H.
通讯作者：
Sherman, David H.
DOI：
10.1073/pnas.0806305105
发表时间：
2008-09-02
影响因子：
11.1
作者：
Calderone, Christopher T.;Bumpus, Stefanie B.;Magarvey, Nathan A.
通讯作者：
Magarvey, Nathan A.
影响因子：
3.2
作者：
Caffrey, P
通讯作者：
Caffrey, P
影响因子：
3.2
作者：
Beck, Zachary Q.;Burr, Douglas A.;Sherman, David H.
通讯作者：
Sherman, David H.
影响因子：
3.5
作者：
Kerksiek, K;Mejillano, MR;Himes, RH
通讯作者：
Himes, RH