Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents
The entry of proteins through the cell membrane is challenging, thus limiting their use as potential therapeutics. Seven cell-penetrating peptides, designed in our laboratory, were evaluated for the delivery of proteins. Fmoc solid-phase peptide synthesis was utilized for the synthesis of seven cycl...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Pharmaceuticals |
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| Online Access: | https://www.mdpi.com/1424-8247/16/3/469 |
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| author | Jonathan Moreno Khalid Zoghebi David Salehi Lois Kim Sorour Khayyatnejad Shoushtari Rakesh K. Tiwari Keykavous Parang |
| author_facet | Jonathan Moreno Khalid Zoghebi David Salehi Lois Kim Sorour Khayyatnejad Shoushtari Rakesh K. Tiwari Keykavous Parang |
| author_sort | Jonathan Moreno |
| collection | DOAJ |
| description | The entry of proteins through the cell membrane is challenging, thus limiting their use as potential therapeutics. Seven cell-penetrating peptides, designed in our laboratory, were evaluated for the delivery of proteins. Fmoc solid-phase peptide synthesis was utilized for the synthesis of seven cyclic or hybrid cyclic–linear amphiphilic peptides composed of hydrophobic (tryptophan (W) or 3,3-diphenylalanine (Dip) and positively-charged arginine (R) residues, such as [WR]<sub>4</sub>, [WR]<sub>9</sub>, [WWRR]<sub>4</sub>, [WWRR]<sub>5</sub>, [(RW)<sub>5</sub>K](RW)<sub>5</sub>, [R<sub>5</sub>K]W<sub>7</sub>, and [DipR]<sub>5</sub>. Confocal microscopy was used to screen the peptides as a protein delivery system of model cargo proteins, green and red fluorescein proteins (GFP and RFP). Based on the confocal microscopy results, [WR]<sub>9</sub> and [DipR]<sub>5</sub> were found to be more efficient among all the peptides and were selected for further studies. [WR]<sub>9</sub> (1–10 µM) + protein (GFP and RFP) physical mixture did not show high cytotoxicity (>90% viability) in triple-negative breast cancer cells (MDA-MB-231) after 24 h, while [DipR]<sub>5</sub> (1–10 µM) physical mixture with GFP exhibited more than 81% cell viability. Confocal microscopy images revealed internalization of GFP and RFP in MDA-MB-231 cells using [WR]<sub>9</sub> (2–10 μM) and [DipR]<sub>5</sub> (1–10 µM). Fluorescence-activated cell sorting (FACS) analysis indicated that the cellular uptake of GFP was concentration-dependent in the presence of [WR]<sub>9</sub> in MDA-MB-231 cells after 3 h of incubation at 37 °C. The concentration-dependent uptake of GFP and RFP was also observed in the presence of [DipR<sub>5</sub>] in SK-OV-3 and MDA-MB-231 cells after 3 h of incubation at 37 °C. FACS analysis indicated that the cellular uptake of GFP in the presence of [WR]<sub>9</sub> was partially decreased by methyl-β-cyclodextrin and nystatin as endocytosis inhibitors after 3 h of incubation in MDA-MB-231 cells, whereas nystatin and chlorpromazine as endocytosis inhibitors slightly reduced the uptake of GFP in the presence of [DipR]<sub>5</sub> after 3 h of incubation in MDA-MB-231. [WR]<sub>9</sub> was able to deliver therapeutically relevant proteins (Histone H2A) at different concentrations. These results provide insight into the use of amphiphilic cyclic peptides in the delivery of protein-related therapeutics. |
| first_indexed | 2024-03-11T06:02:22Z |
| format | Article |
| id | doaj.art-8ed404d83c3d4e97a568fff702784685 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-11T06:02:22Z |
| publishDate | 2023-03-01 |
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| series | Pharmaceuticals |
| spelling | doaj.art-8ed404d83c3d4e97a568fff7027846852023-11-17T13:13:15ZengMDPI AGPharmaceuticals1424-82472023-03-0116346910.3390/ph16030469Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery AgentsJonathan Moreno0Khalid Zoghebi1David Salehi2Lois Kim3Sorour Khayyatnejad Shoushtari4Rakesh K. Tiwari5Keykavous Parang6Center for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USACenter for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USACenter for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USACenter for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USACenter for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USACenter for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USACenter for Targeted Drug Delivery, Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Harry and Diane Rinker Health Science Campus, Irvine, CA 92618, USAThe entry of proteins through the cell membrane is challenging, thus limiting their use as potential therapeutics. Seven cell-penetrating peptides, designed in our laboratory, were evaluated for the delivery of proteins. Fmoc solid-phase peptide synthesis was utilized for the synthesis of seven cyclic or hybrid cyclic–linear amphiphilic peptides composed of hydrophobic (tryptophan (W) or 3,3-diphenylalanine (Dip) and positively-charged arginine (R) residues, such as [WR]<sub>4</sub>, [WR]<sub>9</sub>, [WWRR]<sub>4</sub>, [WWRR]<sub>5</sub>, [(RW)<sub>5</sub>K](RW)<sub>5</sub>, [R<sub>5</sub>K]W<sub>7</sub>, and [DipR]<sub>5</sub>. Confocal microscopy was used to screen the peptides as a protein delivery system of model cargo proteins, green and red fluorescein proteins (GFP and RFP). Based on the confocal microscopy results, [WR]<sub>9</sub> and [DipR]<sub>5</sub> were found to be more efficient among all the peptides and were selected for further studies. [WR]<sub>9</sub> (1–10 µM) + protein (GFP and RFP) physical mixture did not show high cytotoxicity (>90% viability) in triple-negative breast cancer cells (MDA-MB-231) after 24 h, while [DipR]<sub>5</sub> (1–10 µM) physical mixture with GFP exhibited more than 81% cell viability. Confocal microscopy images revealed internalization of GFP and RFP in MDA-MB-231 cells using [WR]<sub>9</sub> (2–10 μM) and [DipR]<sub>5</sub> (1–10 µM). Fluorescence-activated cell sorting (FACS) analysis indicated that the cellular uptake of GFP was concentration-dependent in the presence of [WR]<sub>9</sub> in MDA-MB-231 cells after 3 h of incubation at 37 °C. The concentration-dependent uptake of GFP and RFP was also observed in the presence of [DipR<sub>5</sub>] in SK-OV-3 and MDA-MB-231 cells after 3 h of incubation at 37 °C. FACS analysis indicated that the cellular uptake of GFP in the presence of [WR]<sub>9</sub> was partially decreased by methyl-β-cyclodextrin and nystatin as endocytosis inhibitors after 3 h of incubation in MDA-MB-231 cells, whereas nystatin and chlorpromazine as endocytosis inhibitors slightly reduced the uptake of GFP in the presence of [DipR]<sub>5</sub> after 3 h of incubation in MDA-MB-231. [WR]<sub>9</sub> was able to deliver therapeutically relevant proteins (Histone H2A) at different concentrations. These results provide insight into the use of amphiphilic cyclic peptides in the delivery of protein-related therapeutics.https://www.mdpi.com/1424-8247/16/3/469amphiphiliccyclic peptidesintracellular transportationprotein delivery |
| spellingShingle | Jonathan Moreno Khalid Zoghebi David Salehi Lois Kim Sorour Khayyatnejad Shoushtari Rakesh K. Tiwari Keykavous Parang Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents Pharmaceuticals amphiphilic cyclic peptides intracellular transportation protein delivery |
| title | Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents |
| title_full | Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents |
| title_fullStr | Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents |
| title_full_unstemmed | Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents |
| title_short | Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents |
| title_sort | amphiphilic cell penetrating peptides containing arginine and hydrophobic residues as protein delivery agents |
| topic | amphiphilic cyclic peptides intracellular transportation protein delivery |
| url | https://www.mdpi.com/1424-8247/16/3/469 |
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