Targeting HIV-1 Protease Autoprocessing for High-throughput Drug Discovery and Drug Resistance Assessment

Huang, Lin; Li, Linfeng; Tien, ChihFeng; LaBarbera, Daniel V.; Chen, Chaoping

doi:10.1038/s41598-018-36730-4

Cited by 10 publications

(12 citation statements)

References 50 publications

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“…Although this finding could potentially indicate that M pro mediates a hypothetical link between COVID-19 and hematological disorders, like virus induced porphyria [ 30 ] and SARS-Cov-2 induced coagulation disorders [ 31 , 32 ] the evidence presented here is far too preliminary and speculative. Furthermore, both substances have been described as being so called promiscuous compounds in HTS [ 33 ]. In this sense, hematoporphyrin could catalyze as a photosensitizer through free radical generation inactivation by oxidation of the catalytic cysteine of M pro [ 34 , 35 ].…”

Section: Resultsmentioning

confidence: 99%

Biochemical screening for SARS-CoV-2 main protease inhibitors

et al. 2020

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The Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) pandemic represents a global challenge. SARS-CoV-2's ability to replicate in host cells relies on the action of its non-structural proteins, like its main protease (M pro). This cysteine protease acts by processing the viruses' precursor polyproteins. As proteases, together with polymerases, are main targets of antiviral drug design, we here have performed biochemical high throughput screening (HTS) with recombinantly expressed SARS-CoV-2 M pro. A fluorescent assay was used to identify inhibitors in a compound library containing known drugs, bioactive molecules and natural products. These screens led to the identification of 13 inhibitors with IC 50 values ranging from 0.2 μM to 23 μM. The screens confirmed several known SARS-CoV M pro inhibitors as inhibitors of SARS-CoV-2 M pro , such as the organo-mercuric compounds thimerosal and phenylmercuric acetate. Benzophenone derivatives could also be identified among the most potent screening hits. Additionally, Evans blue, a sulfonic acid-containing dye, could be identified as an M pro inhibitor. The obtained compounds could be of interest as lead compounds for the development of future SARS-CoV-2 drugs.

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Section: Resultsmentioning

confidence: 99%

Biochemical screening for SARS-CoV-2 main protease inhibitors

et al. 2020

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“…If an acceptor bead is in close proximity to the singlet oxygen, it emits a luminescent signal at 615 nm (and when this proximity is perturbed, there is a loss of signal). As AlphaLISA is amenable to quantitative high-throughput sceeening (qHTS), and can leverage multiple tagged ACE2 and Spike RBD constructs that are commercially available, this methodology could prove useful as a discovery approach for small or large molecules that inhibit infection 15,16 .…”

Section: Introductionmentioning

confidence: 99%

Targeting ACE2-RBD interaction as a platform for COVID19 therapeutics: Development and drug repurposing screen of an AlphaLISA proximity assay

Hanson

Wilson

Shen

et al. 2020

Preprint

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The COVID-19 pandemic, caused by SARS-CoV-2, is a pressing public health emergency garnering rapid response from scientists across the globe. Host cell invasion is initiated through direct binding of the viral spike protein to the host receptor angiotensin-converting enzyme 2 (ACE2). Disrupting the spike-ACE2 interaction is a potential therapeutic target for treating COVID-19. We have developed a proximitybased AlphaLISA assay to measure binding of SARS-CoV-2 spike protein Receptor Binding Domain (RBD) to ACE2. Utilizing this assay platform, a drug-repurposing screen against 3,384 small molecule drugs and pre-clinical compounds was performed, yielding 25 high-quality, small-molecule hits that can be evaluated in cell-based models.This established AlphaLISA RBD-ACE2 platform can facilitate evaluation of biologics or small molecules that can perturb this essential viral-host interaction to further the development of interventions to address the global health pandemic.

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“…Multimerization of polyproteins residues along with complex binding of the viral genomic RNA and other viral proteins in the host cell system gives rise to the formation of new virion particles [7]. e internalization of the newly formed virus is typically triggered by structural protein matrix A (MA), capsid (CA), and nucleocapsid (NC); this includes other spacer peptides p1, p2, and p6 at the virus catalytic site [8].…”

Section: Introductionmentioning

confidence: 99%

“…Without HIV-1 protease, the polyproteins would not be able to cleave into functional products and ultimately would not produce viruses from a host cell nor cause any infection. erefore, due to the fundamental importance of HIV-1 protease for the realization of HIV, it has become the main target in the development of anti-HIV drug therapy [7].…”

Section: Introductionmentioning

confidence: 99%

Inhibition of HIV‐1 Protease by Carpobrotus edulis (L.)

Omoruyi

Ighodaro

Afolayan

et al. 2020

Evidence-Based Complementary and Alternative Medicine

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Carpobrotus edulis (L.) is a plant commonly found in the Eastern Cape Province of South Africa and is used for the general treatment of infections relating to the human immunodeficiency virus (HIV). HIV-1 protease plays an important role during HIV replication and maturation to its infectious form, and therefore inhibition of the enzyme is one of the main focus areas in drug development. The inhibitory effect of a water extract of C. edulis leaves against HIV-1 protease activity was determined using the SensoLyte® 520 HIV-1 protease assay fluorimetric kit and employing a HiLyte Fluor™488/QXL™520 fluorescence resonance energy transfer (FRET) peptide. Cytotoxicity of the extract towards HeLa Chang cell lines was determined using an in vitro MTT assay, and the phytochemical profile of the extract was determined with FT-IR and LC-MS. HIV-1 protease activity was inhibited 83.06% (IC50 1.6 mg/ml) (p<0.0001) by the pepstatin A inhibitor control. Treatment with all C. edulis extract concentrations (16, 1.6, 0.16, and 0.016 mg/ml) inhibited HIV-1 protease activity significantly (p<0.0001) in a typical dose response manner. With regards to cytotoxicity, the negative controls containing untreated HeLa Chang cells exhibited high formazan formation rates in contrast with the positive controls, containing curcumin, which reduced formazan formation significantly (p<0.001), exhibiting cytotoxicity towards the cells. There was no significant (p>0.05) difference in the formazan formation rates between the negative controls and 1, 0.5, 0.125, 0.065, 0.031, and 0.015 mg/ml plant extract, confirming no toxicity of C. edulis extracts towards HeLa Chang cells. Major functional phytochemical compounds identified included alcohols, phenols, alkanes, amines, carboxylic acids, and esters. LC-ESI-TOF/MS analysis revealed the putative identities of main compounds present in the aqueous leaves extract, including some that contribute to its anti-HIV-1 protease action.

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Targeting HIV-1 Protease Autoprocessing for High-throughput Drug Discovery and Drug Resistance Assessment

Cited by 10 publications

References 50 publications

Biochemical screening for SARS-CoV-2 main protease inhibitors

Biochemical screening for SARS-CoV-2 main protease inhibitors

Targeting ACE2-RBD interaction as a platform for COVID19 therapeutics: Development and drug repurposing screen of an AlphaLISA proximity assay

Inhibition of HIV‐1 Protease by Carpobrotus edulis (L.)

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