Targeted covalent inhibitors of MDM2 using electrophile-bearing stapled peptides

Charoenpattarapreeda, Jiraborrirak; Tan, Yaw Sing; Iegre, Jessica; Walsh, Stephen J.; Fowler, Elaine; Eapen, Rohan; Wu, Yinghai; Sore, Hannah F.; Verma, Chandra; Itzhaki, Laura S.; Spring, David R.

doi:10.1039/c9cc04022f

Cited by 25 publications

(25 citation statements)

References 52 publications

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“…Stapled peptides are typically derived from the α-helix from the binding interface, and they are locked into bioactive conformations through the use of chemical linkers. Stapling enhances α-helicity of unstructured short peptides in solution, improves resistance against proteolytic digestion, as well as potency, and often improves cell penetration [15][16][17][18][19][20][21] . One of these stapled peptides, ALRN-6924, is in various clinical trials for the treatment of lymphomas (Phase 1b/2), peripheral T-cell lymphoma (Phase 2a), acute myeloid lymphoma (AML, Phase 1), and advanced myelodysplastic syndrome (MDS, Phase 1) 22 .…”

Section: Introductionmentioning

confidence: 99%

Stapled peptides based on Human Angiotensin-Converting Enzyme 2 (ACE2) potently inhibit SARS-CoV-2 infection in vitro

Curreli

Victor

Ahmed

et al. 2020

Preprint

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Human angiotensin-converting enzyme 2 (ACE2) is the primary receptor of SARS-CoV-2 to enter the host cells and start the infection process. Therefore, it is prudent to design therapeutics based on the critical binding region of ACE2, which is a ~30 aa long helix with a kink in the middle. However, the small peptide in solution may lose its helical conformation and subsequently lose its binding potential to the SARS-CoV-2 RBD, which it utilizes to bind to that helical region. Here we report the design of four stapled peptides based on that helix, which is expected to bind to SARS-CoV-2 with high affinity and prevent the binding of the virus to the ACE2 receptor and disrupt the infection. All stapled peptides showed high helical contents (50 - 94% helicity). On the contrary, the linear control peptide NYBSP-C showed no helicity (19%). We have evaluated the peptides in a pseudovirus based single-cycle assay in HT1080 and human lung cells, A549. Three of the four stapled peptides showed potent antiviral activity in HT1080 (IC50: 1.9 – 4.1 μ) and A549 cells (IC50: 2.2 – 2.8 μ). It is noteworthy that the stapled peptide, NYBSP-3, which showed the least helical content, also had the lowest antiviral activity in both cell lines. The linear peptides NYBSP-C and SBP1, reported recently to bind SARS-CoV-2 with KD of ~47nM affinity, showed no antiviral activity. Most significantly, none of the stapled peptides show any appreciable cytotoxicity at the highest dose tested. We determined the proteolytic stability of one of the most active stapled peptides, NYBSP-4, in human plasma, which showed a half-life (T1/2) of >289 min.

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Section: Introductionmentioning

confidence: 99%

Stapled peptides based on Human Angiotensin-Converting Enzyme 2 (ACE2) potently inhibit SARS-CoV-2 infection in vitro

Curreli

Victor

Ahmed

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Stapled peptides are typically derived from the a-helix from the binding interface, and they are locked into bioactive conformations through the use of chemical linkers. Stapling enhances a-helicity of unstructured short peptides in solution, improves resistance against proteolytic digestion, as well as potency, and often improves cell penetration (15)(16)(17)(18)(19)(20)(21). One of these stapled peptides, ALRN-6924, is in various clinical trials for the treatment of lymphomas (phase 1b/2), peripheral T-cell lymphoma (phase 2a), acute myeloid lymphoma (AML, phase 1), and advanced myelodysplastic syndrome (MDS, phase 1) (22).…”

mentioning

confidence: 99%

Stapled Peptides Based on Human Angiotensin-Converting Enzyme 2 (ACE2) Potently Inhibit SARS-CoV-2 Infection In Vitro

Curreli

Victor

Ahmed

et al. 2020

mBio

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SARS-CoV-2 uses human angiotensin-converting enzyme 2 (ACE2) as the primary receptor to enter host cells and initiate the infection. The critical binding region of ACE2 is an ∼30-amino-acid (aa)-long helix. Here, we report the design of four stapled peptides based on the ACE2 helix, which is expected to bind to SARS-CoV-2 and prevent the binding of the virus to the ACE2 receptor and disrupt the infection. All stapled peptides showed high helical contents (50 to 94% helicity). In contrast, the linear control peptide NYBSP-C showed no helicity (19%). We have evaluated the peptides in a pseudovirus-based single-cycle assay in HT1080/ACE2 cells and human lung cell line A549/ACE2, overexpressing ACE2. Three of the four stapled peptides showed potent antiviral activity in HT1080/ACE2 (50% inhibitory concentration [IC50]: 1.9 to 4.1 μM) and A549/ACE2 (IC50: 2.2 to 2.8 μM) cells. The linear peptide NYBSP-C and the double-stapled peptide StRIP16, used as controls, showed no antiviral activity. Most significantly, none of the stapled peptides show any cytotoxicity at the highest dose tested. We also evaluated the antiviral activity of the peptides by infecting Vero E6 cells with the replication-competent authentic SARS-CoV-2 (US_WA-1/2020). NYBSP-1 was the most efficient, preventing the complete formation of cytopathic effects (CPEs) at an IC100 of 17.2 μM. NYBSP-2 and NYBSP-4 also prevented the formation of the virus-induced CPE with an IC100 of about 33 μM. We determined the proteolytic stability of one of the most active stapled peptides, NYBSP-4, in human plasma, which showed a half-life (T1/2) of >289 min. IMPORTANCE SARS-CoV-2 is a novel virus with many unknowns. No vaccine or specific therapy is available yet to prevent and treat this deadly virus. Therefore, there is an urgent need to develop novel therapeutics. Structural studies revealed critical interactions between the binding site helix of the ACE2 receptor and SARS-CoV-2 receptor-binding domain (RBD). Therefore, targeting the entry pathway of SARS-CoV-2 is ideal for both prevention and treatment as it blocks the first step of the viral life cycle. We report the design of four double-stapled peptides, three of which showed potent antiviral activity in HT1080/ACE2 cells and human lung carcinoma cells, A549/ACE2. Most significantly, the active stapled peptides with antiviral activity against SARS-CoV-2 showed high α-helicity (60 to 94%). The most active stapled peptide, NYBSP-4, showed substantial resistance to degradation by proteolytic enzymes in human plasma. The lead stapled peptides are expected to pave the way for further optimization of a clinical candidate.

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“…In recent years, a number of covalent PPI inhibitors have been approved by the US Food and Drug Administration (FDA) ( Table 1). In the specific field of oncology, one covalent PPI inhibitor has been approved by the FDA, while others have entered clinical trials [84,89,[91][92][93]. One recent example is the covalent inhibitor COH000, a highly specific allosteric inhibitor of the SUMO (small ubiquitin-related modifier) E1 enzyme, an anticancer target [88].…”

Section: Successful Covalent Ppi Inhibitors In Cancer Therapymentioning

confidence: 99%

The design and development of covalent protein-protein interaction inhibitors for cancer treatment

et al. 2020

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Protein-protein interactions (PPIs) are central to a variety of biological processes, and their dysfunction is implicated in the pathogenesis of a range of human diseases, including cancer. Hence, the inhibition of PPIs has attracted significant attention in drug discovery. Covalent inhibitors have been reported to achieve high efficiency through forming covalent bonds with cysteine or other nucleophilic residues in the target protein. Evidence suggests that there is a reduced risk for the development of drug resistance against covalent drugs, which is a major challenge in areas such as oncology and infectious diseases. Recent improvements in structural biology and chemical reactivity have enabled the design and development of potent and selective covalent PPI inhibitors. In this review, we will highlight the design and development of therapeutic agents targeting PPIs for cancer therapy.

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Targeted covalent inhibitors of MDM2 using electrophile-bearing stapled peptides

Abstract: An electrophile-bearing linker was developed for the generation of a stapled peptide covalent inhibitor of MDM2 via a two-component CuAAC peptide stapling.

Cited by 25 publications

References 52 publications

Stapled peptides based on Human Angiotensin-Converting Enzyme 2 (ACE2) potently inhibit SARS-CoV-2 infection in vitro

Stapled peptides based on Human Angiotensin-Converting Enzyme 2 (ACE2) potently inhibit SARS-CoV-2 infection in vitro

Stapled Peptides Based on Human Angiotensin-Converting Enzyme 2 (ACE2) Potently Inhibit SARS-CoV-2 Infection In Vitro

The design and development of covalent protein-protein interaction inhibitors for cancer treatment

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