Targeting the Hemopexin-like Domain of Latent Matrix Metalloproteinase-9 (proMMP-9) with a Small Molecule Inhibitor Prevents the Formation of Focal Adhesion Junctions.
Targeting the Hemopexin-like Domain of Latent Matrix Metalloproteinase-9 (proMMP-9) with a Small Molecule Inhibitor Prevents the Formation of Focal Adhesion Junctions.
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DOI：
10.1021/acschembio.7b00758
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发表时间：
2017-11-17
影响因子：
4
通讯作者：
Sampson NS
中科院分区：
文献类型：
--
作者：
Alford VM;Kamath A;Ren X;Kumar K;Gan Q;Awwa M;Tong M;Seeliger MA;Cao J;Ojima I;Sampson NS
靶特异性的缺乏极大地阻碍了针对基质金属蛋白酶（MMP）的抑制剂开发用于治疗各种癌症的成功。MMP催化结构域是高度保守的，而MMP的血红素样结构域对于每个家族成员是独特的。MMP-9的血红素蛋白样结构域通过与细胞表面蛋白（包括CD 44和α4β1整联蛋白）的自身相互作用和异源相互作用增强癌细胞迁移。这些相互作用激活EGFR-MAP激酶依赖性信号传导，导致细胞迁移。在这项工作中，我们基于命中分子N-[4-（二氟甲氧基）苯基]-2-[（4-氧代-6-丙基-1H-嘧啶-2-基）硫基]-乙酰胺产生了靶向MMP-9的血红素样结构域的化合物库。我们鉴定N-（4-氟苯基）-4-（4-氧代-3，4，5，6，7，8-六氢喹唑啉-2-基硫基）丁酰胺3c为特异性结合proMMP-9血红素样结构域的有效先导化合物（Kd = 320 nM）。我们证明，MMP-9同源二聚化的3c破坏阻止了proMMP-9与α4β1整联蛋白和CD 44的结合，并导致EGFR的解离。这种破坏导致Src及其下游靶蛋白粘着斑激酶（FAK）和桩蛋白（PAX）的磷酸化减少，这些蛋白与促进肿瘤细胞生长、迁移和侵袭有关。使用鸡绒毛膜尿囊膜在体内测定，我们证明，500 nM的3C块癌细胞的基底膜的入侵，并减少血管生成。总之，我们提出了3c的作用机制，其中靶向血红素结合蛋白结构域通过同时破坏α4β1整合素和EGFR信号传导途径导致癌细胞迁移减少，从而防止信号旁路。通过血红素样结构域靶向是抗转移药物开发的有力方法。
A lack of target specificity has greatly hindered the success of inhibitor development against matrix metalloproteinases (MMPs) for the treatment of various cancers. The MMP catalytic domains are highly conserved, whereas the hemopexin-like domains of MMPs are unique to each family member. The hemopexin-like domain of MMP-9 enhances cancer cell migration through self-interaction and heterointeractions with cell surface proteins including CD44 and α4β1 integrin. These interactions activate EGFR-MAP kinase dependent signaling that leads to cell migration. In this work, we generated a library of compounds, based on hit molecule N-[4-(difluoromethoxy)phenyl]-2-[(4-oxo-6-propyl-1H-pyrimidin-2-yl)sulfanyl]-acetamide, that target the hemopexin-like domain of MMP-9. We identify N-(4-fluorophenyl)-4-(4-oxo-3,4,5,6,7,8-hexahydroquinazolin-2-ylthio)butanamide, 3c, as a potent lead (Kd = 320 nM) that is specific for binding to the proMMP-9 hemopexin-like domain. We demonstrate that 3c disruption of MMP-9 homodimerization prevents association of proMMP-9 with both α4β1 integrin and CD44 and results in the dissociation of EGFR. This disruption results in decreased phosphorylation of Src and its downstream target proteins focal adhesion kinase (FAK) and paxillin (PAX), which are implicated in promoting tumor cell growth, migration, and invasion. Using a chicken chorioallantoic membrane in vivo assay, we demonstrate that 500 nM 3c blocks cancer cell invasion of the basement membrane and reduces angiogenesis. In conclusion, we present a mechanism of action for 3c whereby targeting the hemopexin domain results in decreased cancer cell migration through simultaneous disruption of α4β1 integrin and EGFR signaling pathways, thereby preventing signaling bypass. Targeting through the hemopexin-like domain is a powerful approach to antimetastatic drug development.
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影响因子：
6.8
作者：
Cathcart, Jillian;Pulkoski-Gross, Ashleigh;Cao, Jian
通讯作者：
Cao, Jian
影响因子：
8.4
作者：
Curran, S;Murray, GI
通讯作者：
Murray, GI
影响因子：
3.7
作者：
Chou, Jonathan;Chan, Matilda F.;Werb, Zena
通讯作者：
Werb, Zena
DOI：
10.1007/978-1-4939-3444-7_15
发表时间：
2016-01-01
期刊：
BREAST CANCER: METHODS AND PROTOCOLS
影响因子：
--
作者：
Alford, Vincent M.;Roth, Eric;Cao, Jian
通讯作者：
Cao, Jian
影响因子：
14.8
作者：
Imai, Kazushi;Okada, Yasunori
通讯作者：
Okada, Yasunori