We have synthesized (CPC Scientific, Inc.) four stapled peptides, as depicted in Figure 2. We also synthesized the linear peptide, NYBSP-C, as a control. Besides, we purchased a linear peptide, SBP1, to use as a control, which was reported recently to bind to SARS-CoV-2 RBD with high affinity (KD = 47nM).
Abstract
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 µM) and A549 cells (IC50: 2.2 – 2.8 µM). 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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Boehm, M., Beaumont, K., Jones, R.M., Kalgutkar, A.S., Zhang, L., Atkinson, K., Bai, G., Brown, J.A., Eng, H., Goetz, G.H. and Holder, B.R. Journal of Medicinal Chemistry (2017).
- Pfizer Worldwide Research & Development, Cambridge, Massachusetts 02139, United States
- Pfizer Worldwide Research & Development, Groton, Connecticut 06340, United States
The chemokine receptor CXCR7 is an attractive target for a variety of diseases. While several small-molecule modulators of CXCR7 have been reported, peptidic macrocycles may provide advantages in terms of potency, selectivity, and reduced off-target activity
Bainbridge, Travis W., et al. Scientific Reports 7.1 (2017): 12524.
- Protein Chemistry, Molecular Oncology, Discovery Chemistry, Cancer Immunology, and Neuroscience. Genentech Inc., South San Francisco, CA, 94080, USA
"ANP FAP and aP NIRF were synthesized by CPC Scientific (Sunnyvale, CA)… Another internally-quenched FRET peptide substrate (ANPFAP) for FAP has recently been reported and demonstrated for use as an activity-based, in vivo imaging tool. The peptide sequence contains two internal Gly-Pro dipeptide motifs, susceptible to FAP cleavage and a Cy5.5/QSY21, quenched-FRET pair."
Coorens, Maarten, et al. The Journal of Immunology 199.4 (2017): 1418-1428.
"CATH-2 and LL-37 were synthesized by Fmoc-chemistry at CPC Scientific (Sunnyvale, CA)."
Tcholakov, I., Grimshaw, C.E., Shi, L., Kiryanov, A., Murphy, S.T., Larson, C.J., Plonowski, A. and Ermolieff, J. Bioscience Reports 37, no. 3 (2017): BSR20170275.
- Departments of In Vitro Pharmacology, Immunology, Enzymology and Biophysical Chemistry, Medicinal Chemistry, External Innovation, Metabolic Disease, In Vitro Pharmacology, and Gastrointestinal and Enterology Discovery Unit, Takeda California, Inc., 10410 Science Center Drive, San Diego, CA 92121, U.S.A
The substrate used for our PHD2 kinetic study was a 17-mer peptide mimicking the sequence of HIF-1a surrounding the Pro564 residue hydroxylated by the PHD enzymes (Biotin-DLEMLAPYIPMDDDFQL). The substrate used for FIH1 was a 34-mer peptide mimicking the sequence of HIF-1α surrounding residue Asn803 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL). Both peptides were synthesized by CPC Scientific Inc.
Wilson, Sarah S., et al. PLoS Pathogens 13.6 (2017): e1006446.
"Cryptdin 2 (UniProtKB: P28309, LRDLVCYCRTRGCKRRERMNGTCRKGHLMYTLCCR)... obtained by oxidative refolding of partially purified linear peptides (synthesized by CPC Scientific...) and purifying the correctly folded species by reverse-phase high-pressure liquid chromatography (RP-HPLC). Purity was determined by analytical RP-HPLC, and the mass of the disulfide-bonded peptides was verified by high mass accuracy liquid chromatography-mass spectrometry."
Sattiraju, A., et al. Oncotarget 2017, 8 (26), 42997-43007.
For Ac-225 labeling, the prepared DOTA-Pep-1L (CPC-scientific, San Jose, CA) was incubated with Ac-225 at 70°C for 50 minutes. The TLC plates were scanned on a BioScan Imaging Scanner. Cu-64 was purchased from Washington University in St. Louis. The custom peptide specific to IL13RA2 and a scrambled peptide were conjugated with NOTA by CPC scientific Inc (San Jose, CA). Both the peptides, Pep-1L and scrambled peptide-NOTA were radiolabeled with Cu-64 according to the previously reported methods [15].
Puthenveetil, Sujiet, et al. PloS One 12.5 (2017): e0178452.
- Worldwide Medicinal Chemistry, Pfizer Global R&D, Groton, Connecticut, United States of America.
- Pfizer Oncology Research, Pearl River, NY, United States of America.
"Peptide-payload conjugate (7) was prepared by reacting 2mM aizoacetyl-Ser-Lys-Gly-Ser-Lys (CPC scientific, inc. Sunnywale, CA) with 8 mM NHS-ester payload [19] in 50% Dimethyl sulfoxide (DMSO), 50 mM borate buffer pH 8.5 for 2 h at 37°C."
Kwon, Ester J., et al. Advanced Materials 29.35 (2017).
"[..] synthesized for initial screening with FAM-conjugated lysine at the C-terminal end of the membrane-interactive peptide and with or without D[KLAKLAK]2 on the C-terminus using standard Fmoc chemistry[..] All peptides were synthesized with N-terminal myristic acid and C-terminal amine. [..] larger quantities of the peptides were synthesized by CPC Scientific to 90% purity"
Noland, Cameron L., et al. Proceedings of the National Academy of Sciences 114.30 (2017): E6044-E6053.
- Departments of Structural Biology, Infectious Diseases, Biochemical and Cellular Pharmacology, Pathology, Translational Immunology, and Biomolecular Resource Group, Genentech, Inc., South San Francisco, CA 94080.
"..peptide substrates were synthesized based on the Pal lipoprotein, Pal peptideshort (Pam2Cys-SSNKNGGK-Biotin; CPC Scientific, Inc.).."