Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 50
Filtrar
1.
Sci Total Environ ; 858(Pt 1): 159790, 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36309282

RESUMO

The Chinese Loess Plateau has been the cradle of Chinese civilization and the main human settlement in China for thousands of years, where anthropogenic activities are believed to have deeply eroded natural landscapes. After decades of minimal leopard sighting in forests of northern China, due to serious human interference, we recently discovered that the leopard population is recovering. This finding provides hope for successful biodiversity conservation in human-dominated ecosystems. To understand the mechanism of leopard return into such a highly fragmented landscape, we applied the concept of ecological networks (ENs) to identify key factors promoting leopard restoration and quantify the ecological links among habitats. We first determined the existence of a healthy leopard population in the study area based on the size of its home range and presence of breeding individuals. We then innovatively used the relationship between species richness and top predators to generate ENs, and found that the connectivity of ENs had a significant positive interaction with leopard survival. Our study validates the effectiveness of establishing ecologically connected habitats for leopard protection, and highlights the importance of applying ENs for conservation planning in highly fragmented ecosystems. This study provides a successful case for the protection of top predators in human-dominated landscapes.


Assuntos
Ecossistema , Panthera , Animais , Humanos , Conservação dos Recursos Naturais , Biodiversidade , Florestas
2.
Protein & Cell ; (12): 448-458, 2023.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-982526

RESUMO

The adenosine 5'-triphosphate (ATP)-binding cassette (ABC) transporter, IrtAB, plays a vital role in the replication and viability of Mycobacterium tuberculosis (Mtb), where its function is to import iron-loaded siderophores. Unusually, it adopts the canonical type IV exporter fold. Herein, we report the structure of unliganded Mtb IrtAB and its structure in complex with ATP, ADP, or ATP analogue (AMP-PNP) at resolutions ranging from 2.8 to 3.5 Å. The structure of IrtAB bound ATP-Mg2+ shows a "head-to-tail" dimer of nucleotide-binding domains (NBDs), a closed amphipathic cavity within the transmembrane domains (TMDs), and a metal ion liganded to three histidine residues of IrtA in the cavity. Cryo-electron microscopy (Cryo-EM) structures and ATP hydrolysis assays show that the NBD of IrtA has a higher affinity for nucleotides and increased ATPase activity compared with IrtB. Moreover, the metal ion located in the TM region of IrtA is critical for the stabilization of the conformation of IrtAB during the transport cycle. This study provides a structural basis to explain the ATP-driven conformational changes that occur in IrtAB.


Assuntos
Sideróforos/metabolismo , Ferro/metabolismo , Mycobacterium tuberculosis/metabolismo , Microscopia Crioeletrônica , Trifosfato de Adenosina/metabolismo , Transportadores de Cassetes de Ligação de ATP
3.
Protein & Cell ; (12): 17-27, 2023.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-971604

RESUMO

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (Mpro) and papain like protease (PLpro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851Mpro inhibitors and 205 PLpro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both Mpro and PLpro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of Mpro inhibitors and over 20% of PLpro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 Mpro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.


Assuntos
Humanos , Antivirais/química , COVID-19 , Tratamento Farmacológico da COVID-19 , Ensaios de Triagem em Larga Escala , Simulação de Acoplamento Molecular , Inibidores de Proteases/química , SARS-CoV-2/enzimologia , Proteínas não Estruturais Virais
4.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-499047

RESUMO

Nirmatrelvir, an oral antiviral targeting the 3CL protease of SARS-CoV-2, has been demonstrated to be clinically useful in reducing hospitalization or death due to COVID-191,2. However, as SARS-CoV-2 has evolved to become resistant to other therapeutic modalities3-9, there is a concern that the same could occur for nirmatrelvir. Here, we have examined this possibility by in vitro passaging of SARS-CoV-2 in increasing concentrations of nirmatrelvir using two independent approaches, including one on a large scale in 480 wells. Indeed, highly resistant viruses emerged from both, and their sequences revealed a multitude of 3CL protease mutations. In the experiment done at a larger scale with many replicates, 53 independent viral lineages were selected with mutations observed at 23 different residues of the enzyme. Yet, several common mutational pathways to nirmatrelvir resistance were preferred, with a majority of the viruses descending from T21I, P252L, or T304I as precursor mutations. Construction and analysis of 13 recombinant SARS-CoV-2 clones, each containing a unique mutation or a combination of mutations showed that the above precursor mutations only mediated low-level resistance, whereas greater resistance required accumulation of additional mutations. E166V mutation conferred the strongest resistance (~100-fold), but this mutation resulted in a loss of viral replicative fitness that was restored by compensatory changes such as L50F and T21I. Structural explanations are discussed for some of the mutations that are proximal to the drug-binding site, as well as cross-resistance or lack thereof to ensitrelvir, another clinically important 3CL protease inhibitor. Our findings indicate that SARS-CoV-2 resistance to nirmatrelvir does readily arise via multiple pathways in vitro, and the specific mutations observed herein form a strong foundation from which to study the mechanism of resistance in detail and to inform the design of next generation protease inhibitors.

5.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-489529

RESUMO

Neutralizing antibodies (NAbs) can prevent and treat infections caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, continuously emerging variants, such as Omicron, have significantly reduced the potency of most known NAbs. The selection of NAbs with broad neutralizing activities and the identification of conserved critical epitopes are still urgently needed. Here, we identified an extremely potent antibody (55A8) by single B-cell sorting from convalescent SARS-CoV-2-infected patients that recognized the receptor-binding domain (RBD) in the SARS-CoV-2 spike (S) protein. 55A8 could bind to wild-type SARS-CoV-2, Omicron BA.1 and Omicron BA.2 simultaneously with 58G6, a NAb previously identified by our group. Importantly, an antibody cocktail containing 55A8 and 58G6 (2-cocktail) showed synergetic neutralizing activity with a half-maximal inhibitory concentration (IC50) in the picomolar range in vitro and prophylactic efficacy in hamsters challenged with Omicron (BA.1) through intranasal delivery at an extraordinarily low dosage (25 g of each antibody daily) at 3 days post-infection. Structural analysis by cryo-electron microscopy (cryo-EM) revealed that 55A8 is a Class III NAb that recognizes a highly conserved epitope. It could block angiotensin-converting enzyme 2 (ACE2) binding to the RBD in the S protein trimer via steric hindrance. The epitopes in the RBD recognized by 55A8 and 58G6 were found to be different and complementary, which could explain the synergetic mechanism of these two NAbs. Our findings not only provide a potential antibody cocktail for clinical use against infection with current SARS-CoV-2 strains and future variants but also identify critical epitope information for the development of better antiviral agents.

6.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-928784

RESUMO

BACKGROUND@#Dabrafenib+Trametinib/Dabrafenib targeted therapy has been approved for V-RAF murine sarcoma viral oncogene homolog B1 with amino acid substitution for valine at position 600 (BRAF V600E) in lung cancer patients, however, the targeted therapy strategy for lung cancer patients with BRAF non-V600E mutations has not been determined yet. This study intends to explore the efficacy of targeted therapy for BRAF non-V600E mutant lung cancer, and provide a reference for clinical treatment.@*METHODS@#Computer search of PubMed, Cochrane Library, Embase, Web of Science, Clinicaltrials.gov, CBM, CNKI, Wanfang database. Collect the relevant literature relevant on the targeted therapy of BRAF non-V600E mutant lung cancer, and conduct a descriptive analysis of the included literature.@*RESULTS@#There were 10 articles that met the inclusion criteria, including 3 cohort studies and 7 case reports. 18 patients with BRAF non-V600E mutant lung cancer were ineffective to vermurafenib; 1 patient obtained partial response (PR) after applying vermurafenib, 5 patients did not respond to BRAF inhibitors; 9 patients showed a potential clinical benefit rate of 34% after monotherapy with trametinib; 7 patients have different degrees of benefit from dabrafenib and trametinib on progression-free survival (PFS); 1 patient is effective to sorafenib.@*CONCLUSIONS@#At present, there is no standard treatment specification for BRAF non-V600E mutation targeted therapy. The challenge lies in the heterogeneous mutation of BRAF gene. Different mutation types respond differently to targeted therapy. In addtion, real-world research evidence is scarce, so it is necessary to carry out further large-sample high-quality research to provide reference for clinical practice.


Assuntos
Animais , Humanos , Camundongos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Neoplasias Pulmonares/genética , Mutação , Inibidores de Proteínas Quinases/uso terapêutico , Proteínas Proto-Oncogênicas B-raf/genética
7.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-931355

RESUMO

Based on the online course of Medical Imaging, the online and offline mixed teaching mode is an important reform against the traditional teaching mode. Combining the teaching reform experience of Chongqing Medical University, this paper introduces the resource construction of the online teaching platform of Medical Imaging, the construction of the online and offline mixed teaching mode (including the overall reconstruction design of the online and offline teaching content, and the organic integration of the online and offline teaching content before, during and after class), the details of teaching implementation (taking ischemic cerebral infarction as an example), the teaching efficiency, the existing problems and related improvement measures. The preliminary practice shows that it has great significance in enhancing the teaching quality since it can break through time and space constraints, expand teaching space, effectively stimulate students' learning autonomy, enhance their practical skills, increase the communication and interaction between teachers and students, and improve the teaching effect.

8.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-440481

RESUMO

Accumulating mutations in the SARS-CoV-2 Spike (S) protein can increase the possibility of immune escape, challenging the present COVID-19 prophylaxis and clinical interventions. Here, 3 receptor binding domain (RBD) specific monoclonal antibodies (mAbs), 58G6, 510A5 and 13G9, with high neutralizing potency blocking authentic SARS-CoV-2 virus displayed remarkable efficacy against authentic B.1.351 virus. Each of these 3 mAbs in combination with one neutralizing Ab recognizing non-competing epitope exhibited synergistic effect against authentic SARS-CoV-2 virus. Surprisingly, structural analysis revealed that 58G6 and 13G9, encoded by the IGHV1-58 and the IGKV3-20 germline genes, both recognized the steric region S470-495 on the RBD, overlapping the E484K mutation presented in B.1.351. Also, 58G6 directly bound to another region S450-458 in the RBD. Significantly, 58G6 and 510A5 both demonstrated prophylactic efficacy against authentic SARS-CoV-2 and B.1.351 viruses in the transgenic mice expressing human ACE2 (hACE2), protecting weight loss and reducing virus loads. These 2 ultrapotent neutralizing Abs can be promising candidates to fulfill the urgent needs for the prolonged COVID-19 pandemic.

9.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-434764

RESUMO

The global emergence of SARS-CoV-2 has triggered numerous efforts to develop therapeutic options for COVID-19 pandemic. The main protease of SARS-CoV-2 (Mpro), which is a critical enzyme for transcription and replication of SARS-CoV-2, is a key target for therapeutic development against COVID-19. An organoselenium drug called ebselen has recently been demonstrated to have strong inhibition against Mpro and antiviral activity but its molecular mode of action is unknown preventing further development. We have examined the binding modes of ebselen and its derivative in Mpro via high resolution co-crystallography and investigated their chemical reactivity via mass spectrometry. Stronger Mpro inhibition than ebselen and potent ability to rescue infected cells were observed for a number of ebselen derivatives. A free selenium atom bound with cysteine 145 of Mpro catalytic dyad has been revealed by crystallographic studies of Mpro with ebselen and MR6-31-2 suggesting hydrolysis of the enzyme bound organoselenium covalent adduct, formation of a phenolic by-product is confirmed by mass spectrometry. The target engagement of these compounds with an unprecedented mechanism of SARS-CoV-2 Mpro inhibition suggests wider therapeutic applications of organo-selenium compounds in SARS-CoV-2 and other zoonotic beta-corona viruses.

10.
Eur J Trauma Emerg Surg ; 47(6): 1679-1686, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33029659

RESUMO

OBJECTIVE: Investigating the anatomical characteristics of supra-acetabular screw corridor as well as comparing the biomechanical stability between semi- and full-length screw external fixations for Tile C1 pelvic fractures. METHODS: 50 male and 50 female uninjured pelvic CT data were converted into three-dimensional models and the supra-acetabular corridors were reconstructed by the Mimics software. The horizontal slice was redefined passing through the bilateral anterior inferior iliac spine and bilateral posterior superior iliac spine at the same time. On every horizontal slices, a 5 mm diameter virtual screw was placed along with the unilateral anterior narrowing to posterior narrowing of the supra-acetabular corridor, and the relevant parameters of the semi- and full-length screw fixations were measured from inferior to superior at an interval height of 2.5 mm until the width of the corridor was less than 5 mm. Thus, the insertion position was located, as well as the inclination angle ranges were measured. The same procedure was done on a bilateral supra-acetabular corridor for each patient. Last, a Tile C1 pelvic finite element model was simulated and fixed with either semi- or a full-length anterior supra-acetabular external fixator. A 500 N loading was tested on the model with three directions (cranial-caudal, anterior-posterior and lateral-medial) and the stiffness was determined by the max displacements. RESULTS: The insertion points of supra-acetabular screw for 66% (66/100) of men and 78% (78/100) of women were located at the outer lower part of the anterior inferior iliac spine (AIIS) (p < 0.001). The medial inclination angles of the screw were 29.50 ± 4.05° for men and 28.00 ± 3.78° for women (p = 0.007). The cranial inclination angles of the screw were 29.40 ± 5.38° for men and 28.57 ± 6.01° for women (p = 0.306). The safe inclination angle ranges of the semi-length screw were 3°-6° bigger than the full-length screw. For semi-length corridor, the widths were 14.07 ± 2.39 mm for men and 11.29 ± 1.81 mm for women (p < 0.001) and the depths were 71.77 ± 5.96 mm for men and 69.26 ± 5.21 mm for women (p < 0.001). For full-length corridor, the widths were 10.61 ± 2.10 mm for men and 8.44 ± 2.03 mm for women (p < 0.001) and the depths were 133.37 ± 7.60 mm for men and 129.01 ± 8.12 mm for women (p < 0.001). In the finite element analysis, the max displacements of the pelvis under three loading modes (cranial-caudal, anterior-posterior and lateral-medial) for semi-length fixation were 0.772 mm, 0.409 mm and 0.331 mm and for full-length fixation were 0.727 mm, 0.385 mm and 0.262 mm. CONCLUSION: The outer lower part of AIIS is recommended as an insertion point of the supra-acetabular screw with about 30° medial and cranial inclination angles, and the semi-length screw is safer for placing and has satisfactory biomechanical stability by compared with a full-length screw.


Assuntos
Fixadores Externos , Fixação de Fratura , Acetábulo/diagnóstico por imagem , Acetábulo/cirurgia , Feminino , Análise de Elementos Finitos , Fixação Interna de Fraturas , Humanos , Masculino
11.
Protein & Cell ; (12): 877-888, 2021.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-922482

RESUMO

A new coronavirus (SARS-CoV-2) has been identified as the etiologic agent for the COVID-19 outbreak. Currently, effective treatment options remain very limited for this disease; therefore, there is an urgent need to identify new anti-COVID-19 agents. In this study, we screened over 6,000 compounds that included approved drugs, drug candidates in clinical trials, and pharmacologically active compounds to identify leads that target the SARS-CoV-2 papain-like protease (PLpro). Together with main protease (M


Assuntos
Humanos , Antivirais/uso terapêutico , Sítios de Ligação , COVID-19/virologia , Proteases Semelhantes à Papaína de Coronavírus/metabolismo , Cristalografia por Raios X , Avaliação Pré-Clínica de Medicamentos , Reposicionamento de Medicamentos , Ensaios de Triagem em Larga Escala/métodos , Imidazóis/uso terapêutico , Concentração Inibidora 50 , Simulação de Dinâmica Molecular , Mutagênese Sítio-Dirigida , Naftoquinonas/uso terapêutico , Inibidores de Proteases/uso terapêutico , Estrutura Terciária de Proteína , Proteínas Recombinantes/isolamento & purificação , SARS-CoV-2/isolamento & purificação
12.
Chinese Journal of Geriatrics ; (12): 1295-1298, 2021.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-911006

RESUMO

Objective:To examine the influence of hyperlipidemia on perioperative outcomes of rectal cancer patients.Methods:From June 2016 to June 2019, 86 patients who had received laparotomy or laparoscopic radical resection of rectal cancer at the Department of General Surgery of Nanjing Jiangbei People's Hospital were selected as research subjects.Preoperative blood lipids were measured.Based on lipid levels, patients were divided into the hyperlipidemia group(46 cases)and the normal blood lipid group(40 cases). Operative time, intraoperative blood loss, postoperative recovery outcomes, incidence of anastomotic fistula, and incidences of incision fat liquefaction and incision infections were compared between the two groups.SPSS21.0 was used for data analysis.Results:Compared with the normal lipid group, the operative time of the hyperlipidemia group was significantly longer and the amount of intraoperative blood loss was higher in the hyperlipidemia group, and the differences were statistically significant.Compared with the normal lipid group, the hyperlipidemia group showed a longer time to first postoperative flatus and to first postoperative oral intake, delayed drainage tube removal and ambulation, and longer hospital stays, with statistically significant differences.There were 17 cases of postoperative complications in the hyperlipidemia group(37.0%)and 4 cases in the normal lipid group(10.0%), and the difference was statistically significant( χ2=7.027, P=0.008). Conclusions:Hyperlipidemia has adverse effects on surgical outcomes and postoperative recovery for rectal cancer patients.

13.
Preprint em Inglês | medRxiv | ID: ppmedrxiv-20139931

RESUMO

ObjectiveTo evaluate the diagnostic efficiency of different methods in detecting COVID-19 to provide preliminary evidence on choosing favourable method for COVID-19 detection. MethodsPubMed, Web of Science and Embase databases were searched for identifing eligible articles. All data were calculated utilizing Meta Disc 1.4, Revman 5.3.2 and Stata 12. The diagnostic efficiency was assessed via these indicators including summary sensitivity and specificity, positive likelihood ratio (PLR), negative LR (NLR), diagnostic odds ratio (DOR), summary receiver operating characteristic curve (sROC) and calculate the AUC. Results18 articles (3648 cases) were included. The results showed no significant threshold exist. EPlex: pooled sensitivity was 0.94; specificity was 1.0; PLR was 90.91; NLR was 0.07; DOR was 1409.49; AUC=0.9979, Q*=0.9840. Panther Fusion: pooled sensitivity was 0.99; specificity was 0.98; PLR was 42.46; NLR was 0.02; DOR was 2300.38; AUC=0.9970, Q*=0.9799. Simplexa: pooled sensitivity was 1.0; specificity was 0.97; PLR was 26.67; NLR was 0.01; DOR was 3100.93; AUC=0.9970, Q*=0.9800. Cobas(R): pooled sensitivity was 0.99; specificity was 0.96; PLR was 37.82; NLR was 0.02; DOR was 3754.05; AUC=0.9973, Q*=0.9810. RT-LAMP: pooled sensitivity was 0.98; specificity was 0.99; PLR was 36.22; NLR was 0.04; DOR was 751.24; AUC=0.9905, Q*=0.9596. Xpert Xpress: pooled sensitivity was 0.99; specificity was 0.97; PLR was 27.44; NLR was 0.01; DOR was 3488.15; AUC=0.9977, Q*=0.9829. ConclusionsThese methods (ePlex, Panther Fusion, Simplexa, Cobas(R), RT-LAMP and Xpert Xpress) bear higher sensitivity and specificity, and might be efficient methods complement to the gold standard.

14.
Preprint em Inglês | medRxiv | ID: ppmedrxiv-20118059

RESUMO

ObjectiveThis study aimed to summarize the characteristics of chest CT imaging in Chinese patients with Coronavirus Disease 2019 (COVID-19) to provide reliable evidence for further guiding clinical routine. MethodsPubMed, Embase and Web of Science databases were thoroughly searched to identified relevant articles involving the features of chest CT imaging in Chinese patients with COVID-19. All data were analyzed utilizing R software version i386 4.0.0. Random-effects models were employed to calculate pooled mean differences. Results19 trials incorporating 1332 cases were included in the study. The results demonstrated that the incidence of ground-glass opacities (GGO) was 0.79, consolidation was 0.34; mixed GGO and consolidation was 0.46; air bronchogram sign was 0.41; crazy paving pattern was 0.32; interlobular septal thickening was 0.55; reticulation was 0.30; bronchial wall thickening was 0.24; vascular enlargement was 0.74. subpleural linear opacity was 0.28; intrathoracic lymph node enlargement was 0.03; pleural effusions was 0.03. The distribution in lung: the incidence of central was 0.05; peripheral was 0.74; peripheral involving central was 0.38; diffuse was 0.19; unifocal involvement was 0.09; multifocal involvement was 0.57; unilateral was 0.16; bilateral was 0.83; The incidence of lobes involved (>2) was 0.70; lobes involved ([less double equals]2) was 0.35. ConclusionGGO, vascular enlargement, interlobular septal thickening more frequently occurred in patients with COVID-19. Peripheral, bilateral, involved lobes >2 might be the features of COVID-19 in the distribution aspect. Therefore, based on the aboved features of COVID-19 in chest CT imaging, it might be a promising means for identifying COVID-19.

15.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-033233

RESUMO

The antineoplastic drug Carmofur was shown to inhibit SARS-CoV-2 main protease (Mpro). Here the X-ray crystal structure of Mpro in complex with Carmofur reveals that the carbonyl reactive group of Carmofur is covalently bound to catalytic Cys145, whereas its fatty acid tail occupies the hydrophobic S2 subsite. Carmofur inhibits viral replication in cells (EC50 = 24.30 M) and it is a promising lead compound to develop new antiviral treatment for COVID-19.

16.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-996348

RESUMO

SARS-CoV-2 is the etiological agent responsible for the COVID-19 outbreak in Wuhan. Specific antiviral drug are urgently needed to treat COVID-19 infections. The main protease (Mpro) of SARS-CoV-2 is a key CoV enzyme that plays a pivotal role in mediating viral replication and transcription, which makes it an attractive drug target. In an effort to rapidly discover lead compounds targeting Mpro, two compounds (11a and 11b) were designed and synthesized, both of which exhibited excellent inhibitory activity with an IC50 value of 0.05 M and 0.04 M respectively. Significantly, both compounds exhibited potent anti-SARS-CoV-2 infection activity in a cell-based assay with an EC50 value of 0.42 M and 0.33 M, respectively. The X-ray crystal structures of SARS-CoV-2 Mpro in complex with 11a and 11b were determined at 1.5 [A] resolution, respectively. The crystal structures showed that 11a and 11b are covalent inhibitors, the aldehyde groups of which are bound covalently to Cys145 of Mpro. Both compounds showed good PK properties in vivo, and 11a also exhibited low toxicity which is promising drug leads with clinical potential that merits further studies.

17.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-993386

RESUMO

A novel coronavirus (2019-nCoV) outbreak has caused a global pandemic resulting in tens of thousands of infections and thousands of deaths worldwide. The RNA-dependent RNA polymerase (RdRp, also named nsp12), which catalyzes the synthesis of viral RNA, is a key component of coronaviral replication/transcription machinery and appears to be a primary target for the antiviral drug, remdesivir. Here we report the cryo-EM structure of 2019-nCoV full-length nsp12 in complex with cofactors nsp7 and nsp8 at a resolution of 2.9-[A]. Additional to the conserved architecture of the polymerase core of the viral polymerase family and a nidovirus RdRp-associated nucleotidyltransferase (NiRAN) domain featured in coronaviral RdRp, nsp12 possesses a newly identified {beta}-hairpin domain at its N-terminal. Key residues for viral replication and transcription are observed. A comparative analysis to show how remdesivir binds to this polymerase is also provided. This structure provides insight into the central component of coronaviral replication/transcription machinery and sheds light on the design of new antiviral therapeutics targeting viral RdRp. One Sentence SummaryStructure of 2019-nCov RNA polymerase.

18.
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-964882

RESUMO

A new coronavirus (CoV) identified as COVID-19 virus is the etiological agent responsible for the 2019-2020 viral pneumonia outbreak that commenced in Wuhan1-4. Currently there is no targeted therapeutics and effective treatment options remain very limited. In order to rapidly discover lead compounds for clinical use, we initiated a program of combined structure-assisted drug design, virtual drug screening and high-throughput screening to identify new drug leads that target the COVID-19 virus main protease (Mpro). Mpro is a key CoV enzyme, which plays a pivotal role in mediating viral replication and transcription, making it an attractive drug target for this virus5,6. Here, we identified a mechanism-based inhibitor, N3, by computer-aided drug design and subsequently determined the crystal structure of COVID-19 virus Mpro in complex with this compound. Next, through a combination of structure-based virtual and high-throughput screening, we assayed over 10,000 compounds including approved drugs, drug candidates in clinical trials, and other pharmacologically active compounds as inhibitors of Mpro. Six of these inhibit Mpro with IC50 values ranging from 0.67 to 21.4 M. Ebselen also exhibited promising antiviral activity in cell-based assays. Our results demonstrate the efficacy of this screening strategy, which can lead to the rapid discovery of drug leads with clinical potential in response to new infectious diseases where no specific drugs or vaccines are available.

19.
Protein & Cell ; (12): 505-517, 2020.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-828759

RESUMO

Inhibition of Mycobacterium tuberculosis (Mtb) cell wall assembly is an established strategy for anti-TB chemotherapy. Arabinosyltransferase EmbB, which catalyzes the transfer of arabinose from the donor decaprenyl-phosphate-arabinose (DPA) to its arabinosyl acceptor is an essential enzyme for Mtb cell wall synthesis. Analysis of drug resistance mutations suggests that EmbB is the main target of the front-line anti-TB drug, ethambutol. Herein, we report the cryo-EM structures of Mycobacterium smegmatis EmbB in its "resting state" and DPA-bound "active state". EmbB is a fifteen-transmembrane-spanning protein, assembled as a dimer. Each protomer has an associated acyl-carrier-protein (AcpM) on their cytoplasmic surface. Conformational changes upon DPA binding indicate an asymmetric movement within the EmbB dimer during catalysis. Functional studies have identified critical residues in substrate recognition and catalysis, and demonstrated that ethambutol inhibits transferase activity of EmbB by competing with DPA. The structures represent the first step directed towards a rational approach for anti-TB drug discovery.

20.
Protein & Cell ; (12): 505-517, 2020.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-828595

RESUMO

Inhibition of Mycobacterium tuberculosis (Mtb) cell wall assembly is an established strategy for anti-TB chemotherapy. Arabinosyltransferase EmbB, which catalyzes the transfer of arabinose from the donor decaprenyl-phosphate-arabinose (DPA) to its arabinosyl acceptor is an essential enzyme for Mtb cell wall synthesis. Analysis of drug resistance mutations suggests that EmbB is the main target of the front-line anti-TB drug, ethambutol. Herein, we report the cryo-EM structures of Mycobacterium smegmatis EmbB in its "resting state" and DPA-bound "active state". EmbB is a fifteen-transmembrane-spanning protein, assembled as a dimer. Each protomer has an associated acyl-carrier-protein (AcpM) on their cytoplasmic surface. Conformational changes upon DPA binding indicate an asymmetric movement within the EmbB dimer during catalysis. Functional studies have identified critical residues in substrate recognition and catalysis, and demonstrated that ethambutol inhibits transferase activity of EmbB by competing with DPA. The structures represent the first step directed towards a rational approach for anti-TB drug discovery.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...