Enoxaparin augments alpha-1-antitrypsin inhibition of TMPRSS2, a promising drug combination against COVID-19.
Enoxaparin augments alpha-1-antitrypsin inhibition of TMPRSS2, a promising drug combination against COVID-19.
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DOI：
10.1038/s41598-022-09133-9
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发表时间：
2022-03-25
影响因子：
4.6
通讯作者：
Chan ED
中科院分区：
文献类型：
--
作者：
Bai X;Buckle AM;Vladar EK;Janoff EN;Khare R;Ordway D;Beckham D;Fornis LB;Majluf-Cruz A;Fugit RV;Freed BM;Kim S;Sandhaus RA;Chan ED
细胞表面丝氨酸蛋白酶跨膜蛋白2(TMPRSS2)是SARS-CoV-2病毒进入细胞所必需的。我们确定了带负电荷的肝素是否增强了α-1抗胰蛋白酶(AAT)对TMPRSS2的抑制作用。在高表达TMPRSS2的HEK293T细胞中检测到TMPRSS2的活性。我们用人类冠状病毒229E(HCoV-229E)对原代人呼吸道上皮细胞(HAEc)的感染进行了定量，方法是核衣壳蛋白免疫染色和空斑试验。对肝素-TMPRSS2-AAT三元络合物进行了详细的分子模拟。依诺肝素增强AAT对TMPRSS2活性和HCoV-229E感染hAEc的抑制作用。根据这些发现，详细的分子模拟表明：(I)AAT的反应中心环采用了一种抑制活性构象，而不是TMPRSS2的晶体结构，该结构与外源性(Nafamostat)或内源性(HAI-2)TMPRSS2抑制剂结合；(Ii)带负电荷的肝素在TMPRSS2-AAT界面连接相邻的正电斑块，中和其他排斥力。总之，依诺肝素增强了AAT对TMPRSS2和冠状病毒感染的抑制作用。这种宿主导向疗法不太可能受到SARS-CoV-2突变的影响。此外，鉴于AAT和肝素都具有已知的抗炎活性，这种形式的治疗可能既针对病毒，也针对严重新冠肺炎的过度炎症后果。
The cell surface serine protease Transmembrane Protease 2 (TMPRSS2) is required to cleave the spike protein of SARS-CoV-2 for viral entry into cells. We determined whether negatively-charged heparin enhanced TMPRSS2 inhibition by alpha-1-antitrypsin (AAT). TMPRSS2 activity was determined in HEK293T cells overexpressing TMPRSS2. We quantified infection of primary human airway epithelial cells (hAEc) with human coronavirus 229E (HCoV-229E) by immunostaining for the nucleocapsid protein and by the plaque assay. Detailed molecular modeling was undertaken with the heparin–TMPRSS2–AAT ternary complex. Enoxaparin enhanced AAT inhibition of both TMPRSS2 activity and infection of hAEc with HCoV-229E. Underlying these findings, detailed molecular modeling revealed that: (i) the reactive center loop of AAT adopts an inhibitory-competent conformation compared with the crystal structure of TMPRSS2 bound to an exogenous (nafamostat) or endogenous (HAI-2) TMPRSS2 inhibitor and (ii) negatively-charged heparin bridges adjacent electropositive patches at the TMPRSS2–AAT interface, neutralizing otherwise repulsive forces. In conclusion, enoxaparin enhances AAT inhibition of both TMPRSS2 and coronavirus infection. Such host-directed therapy is less likely to be affected by SARS-CoV-2 mutations. Furthermore, given the known anti-inflammatory activities of both AAT and heparin, this form of treatment may target both the virus and the excessive inflammatory consequences of severe COVID-19.
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影响因子：
7.3
作者：
Bai, Xiyuan;Bai, An;Chan, Edward D.
通讯作者：
Chan, Edward D.
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1164/ajrccm.160.4.9807166
发表时间：
1999-10-01
影响因子：
24.7
作者：
Cantin, AM;Woods, DE
通讯作者：
Woods, DE
影响因子：
4.8
作者：
Dementiev, A;Simonovic, M;Gettins, PGW
通讯作者：
Gettins, PGW
影响因子：
--
作者：
Azouz NP;Klingler AM;Callahan V;Akhrymuk IV;Elez K;Raich L;Henry BM;Benoit JL;Benoit SW;Noé F;Kehn-Hall K;Rothenberg ME
通讯作者：
Rothenberg ME