Stapled Wasp Venom-Derived Oncolytic Peptides with Side Chains Induce Rapid Membrane Lysis and Prolonged Immune Responses in Melanoma

Wu, Ye; Li, Dong; Jiang, Yixin; Jin, Jinmei; Liu, Sanhong; Chen, Lili; Zhang, Hong; Zhou, Yifeng; Chen, Hongzhuan; Nagle, Dale G.; Luan, Xin; Zhang, Guoqing

doi:10.1021/acs.jmedchem.0c02237

Cited by 27 publications

(29 citation statements)

References 51 publications

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“…Further, our toxicity assays ( Figures 3A , B , Table 3 ; Supplementary Figures 16 , 17 ) and previous works ( Chapuis et al, 2012 ; Wu et al, 2021 ) support that changing a charged amino acid to S 5 increases the toxicity of peptide analogs. For peptides with a lower positive charge than the natural anoplin, we observed a dose-dependent reduction in cell viability and increased hemolytic activity.…”

Section: Discussionsupporting

confidence: 87%

“…The staple location sites were selected to reduce toxicity and improve the antibacterial activity of anoplin but at the same time to keep its amphipathic character ( Figure 1A ). Previous research showed that substituting anoplin positively charged amino acids (Lys and Arg) with staples increased its oncological activity in cellular models and mouse tumor models ( Wu et al, 2021 ). Thus, we expected that Lys and Arg modifications would increase anoplin toxicity against eukaryotic cells.…”

Section: Resultsmentioning

confidence: 99%

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Stapled Anoplin as an Antibacterial Agent

et al. 2021

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Anoplin is a linear 10-amino acid amphipathic peptide (Gly-Leu-Leu-Lys-Arg-Ile-Lys-Thr-Leu-Leu-NH2) derived from the venom sac of the solitary wasp. It has broad antimicrobial activity, including an antibacterial one. However, the inhibition of bacterial growth requires several dozen micromolar concentrations of this peptide. Anoplin is positively charged and directly interacts with anionic biological membranes forming an α-helix that disrupts the lipid bilayer. To improve the bactericidal properties of anoplin by stabilizing its helical structure, we designed and synthesized its analogs with hydrocarbon staples. The staple was introduced at two locations resulting in different charges and amphipathicity of the analogs. Circular dichroism studies showed that all modified anoplins adopted an α-helical conformation, both in the buffer and in the presence of membrane mimics. As the helicity of the stapled anoplins increased, their stability in trypsin solution improved. Using the propidium iodide uptake assay in Escherichia coli and Staphylococcus aureus, we confirmed the bacterial membrane disruption by the stapled anoplins. Next, we tested the antimicrobial activity of peptides on a range of Gram-negative and Gram-positive bacteria. Finally, we evaluated peptide hemolytic activity on sheep erythrocytes and cytotoxicity on human embryonic kidney 293 cells. All analogs showed higher antimicrobial activity than unmodified anoplin. Depending on the position of the staple, the peptides were more effective either against Gram-negative or Gram-positive bacteria. Anoplin[5-9], with a lower positive charge and increased hydrophobicity, had higher activity against Gram-positive bacteria but also showed hemolytic and destructive effects on eukaryotic cells. Contrary, anoplin[2-6] with a similar charge and amphipathicity as natural anoplin effectively killed Gram-negative bacteria, also pathogenic drug-resistant strains, without being hemolytic and toxic to eukaryotic cells. Our results showed that anoplin charge, amphipathicity, and location of hydrophobic residues affect the peptide destructive activity on the cell wall, and thus, its antibacterial activity. This means that by manipulating the charge and position of the staple in the sequence, one can manipulate the antimicrobial activity.

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Section: Discussionsupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Stapled Anoplin as an Antibacterial Agent

et al. 2021

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“…MP exhibited potent antitumor activity because of the interaction between its cationic amphipathic α-helical region and tumor cell membranes 17 , 18 . In order to enhance its oncolytic potency and resistance to enzyme degradation, an all-hydrocarbon stapling strategy was used to induce a stable α-helical structure 13 . To identify MP derivative(s) with an optimal therapeutic index (TI), we incorporated two ( S )-pentenyl alanine residues (S 5 ) at the i and i + 4 positions of MP to obtain a staple-scanning library by Fmoc solid-phase peptide synthesis (SPPS) 19 , 20 and evaluated their secondary structure features, anti-proliferative, and hemolytic activities (Figure 1 A).…”

Section: Resultsmentioning

confidence: 99%

“…The intrinsic shortcomings of current oncolytic peptides include structural instability, proteolytic susceptibility, and suboptimal antitumor activity. To overcome these drawbacks, we developed a series of wasp venom-derived oncolytic peptides by peptide stapling technology, which significantly improved the α-helical stabilization and anti-tumor effects of parent peptide in B16F10 tumor-bearing mice 13 . However, the requirement for intratumoral administration limits the application of these oncolytic peptides towards both large and metastatic tumors.…”

Section: Introductionmentioning

confidence: 99%

Polymer chimera of stapled oncolytic peptide coupled with anti-PD-L1 peptide boosts immunotherapy of colorectal cancer

Lu¹,

Zhang²,

Zhou³

et al. 2022

Theranostics

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Rationale: Scarce tumor mutation burden and neoantigens create tremendous obstacles for an effective immunotherapy of colorectal cancer (CRC). Oncolytic peptides rise as a promising therapeutic approach that boosts tumor-specific immune responses by inducing antigenic substances. However, the clinical application of oncolytic peptides has been hindered because of structural instability, proteolytic degradation, and undesired toxicity when administered systemically. Methods: Based on wasp venom peptide, an optimized stapled oncolytic peptide MP9 was developed with rigid α-helix, protease-resistance, and CRC cell cytotoxicity. By incorporating four functional motifs that include D-peptidomimetic inhibitor of PD-L1, matrix metalloproteinase-2 (MMP-2) cleavable spacer, and MP9 with 4-arm PEG, a novel peptide-polymer conjugate (PEG-MP9-aPDL1) was obtained and identified as the most promising systemic delivery vehicle with PD-L1 targeting specificity and favorable pharmacokinetic properties. Results: We demonstrated that PEG-MP9-aPDL1-driven oncolysis induces a panel of immunogenic cell death (ICD)-relevant damage-associated molecular patterns (DAMPs) both in vitro and in vivo , which are key elements for immunotherapy with PD-L1 inhibitor. Further, PEG-MP9-aPDL1 exhibited prominent immunotherapeutic efficacy in a CRC mouse model characterized by tumor infiltration of CD8+ T cells and induction of cytotoxic lymphocytes (CTLs) in the spleens. Conclusion: Our findings suggest that PEG-MP9-aPDL1 is an all-in-one platform for oncolytic immunotherapy and immune checkpoint blockade (ICB).

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“…Depending on the peptide structural context, several cyclization strategies can be applied: head-to-tail, side chain-to-head, side chain-to-tail and side chain-to-side chain [5]. A plethora of chemistries have been developed for cyclization, ranging from natural disulfide bridges or isopeptide bonds to metal-catalyzed ring-closing metathesis (RCM), as well as azide-alkyne and Diels-Alder cycloadditions [26,27]. Moreover, peptide conformations may be further constrained by multicyclization and the use of chemical scaffolds to mediate linkage [28].…”

Section: Cyclizationmentioning

confidence: 99%

Protease-Resistant Peptides for Targeting and Intracellular Delivery of Therapeutics

et al. 2021

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Peptides show high promise in the targeting and intracellular delivery of next-generation bio- and nano-therapeutics. However, the proteolytic susceptibility of peptides is one of the major limitations of their activity in biological environments. Numerous strategies have been devised to chemically enhance the resistance of peptides to proteolysis, ranging from N- and C-termini protection to cyclization, and including backbone modification, incorporation of amino acids with non-canonical side chains and conjugation. Since conjugation of nanocarriers or other cargoes to peptides for targeting and cell penetration may already provide some degree of shielding, the question arises about the relevance of using protease-resistant sequences for these applications. Aiming to answer this question, here we provide a critical review on protease-resistant targeting peptides and cell-penetrating peptides (CPPs). Two main approaches have been used on these classes of peptides: enantio/retro-enantio isomerization and cyclization. On one hand, enantio/retro-enantio isomerization has been shown to provide a clear enhancement in peptide efficiency with respect to parent L-amino acid peptides, especially when applied to peptides for drug delivery to the brain. On the other hand, cyclization also clearly increases peptide transport capacity, although contribution from enhanced protease resistance or affinity is often not dissected. Overall, we conclude that although conjugation often offers some degree of protection to proteolysis in targeting peptides and CPPs, modification of peptide sequences to further enhance protease resistance can greatly increase homing and transport efficiency.

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Stapled Wasp Venom-Derived Oncolytic Peptides with Side Chains Induce Rapid Membrane Lysis and Prolonged Immune Responses in Melanoma

Cited by 27 publications

References 51 publications

Stapled Anoplin as an Antibacterial Agent

Stapled Anoplin as an Antibacterial Agent

Polymer chimera of stapled oncolytic peptide coupled with anti-PD-L1 peptide boosts immunotherapy of colorectal cancer

Protease-Resistant Peptides for Targeting and Intracellular Delivery of Therapeutics

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