Selective Integrin Ligands Promote Cell Internalization of the Antineoplastic Agent Fluorouracil

Baiula, Monica; Cirillo, Martina; Martelli, Giulia; Giraldi, Valentina; Elisa, Gasparini; Anelli, Alessandro Claudio; Spampinato, Santi Mario; Giacomini, Daria

doi:10.1021/acsptsci.1c00094

Cited by 7 publications

(15 citation statements)

References 65 publications

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“…Among the previously described small-molecule drug conjugates (SMDC) based on or inspired by peptides, there are only a few conjugates known for targeting the integrins ( Dal Corso et al, 2016 ; Baiula et al, 2021 ; Lerchen et al, 2022 ; Slack et al, 2022 ), particularly α V β 3, with non-peptide homing devices. Such RGD mimetics provide additional possibilities of introducing structural elements and are metabolically more stable than peptides.…”

Section: Resultsmentioning

confidence: 99%

Synthesis and Evaluation of a Non-Peptide Small-Molecule Drug Conjugate Targeting Integrin αVβ3

Paulus

Sewald

2022

Front. Chem.

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An integrin αVβ3-targeting linear RGD mimetic containing a small-molecule drug conjugate (SMDC) was synthesized by combining the antimitotic agent monomethyl auristatin E (MMAE), an enzymatically cleavable Val-Ala-PABC linker with a linear conjugable RGD mimetic. The structure proposal for the conjugable RGD mimetic was suggested upon the DAD mapping analysis of a previously synthesized small-molecule RGD mimetic array based on a tyrosine scaffold. Therefore, a diversifying strategy was developed as well as a novel method for the partial hydrogenation of pyrimidines in the presence of the hydrogenolytically cleavable Cbz group. The small-molecule RGD mimetics were evaluated in an ELISA-like assay, and the structural relationships were analyzed by DAD mapping revealing activity differences induced by structural changes as visualized in dependence on special structural motifs. This provided a lead structure for generation of an SMDC containing the antimitotic drug MMAE. The resulting SMDC containing a linear RGD mimetic was tested in a cell adhesion and an in vitro cell viability assay in comparison to reference SMDCs containing cRGDfK or cRADfK as the homing device. The linear RGD SMDC and the cRGDfK SMDC inhibited adhesion of αVβ3-positive WM115 cells to vitronectin with IC50 values in the low µM range, while no effect was observed for the αVβ3-negative M21-L cell line. The cRADfK SMDC used as a negative control was about 30-fold less active in the cell adhesion assay than the cRGDfK SMDC. Conversely, both the linear RGD SMDC and the cRGDfK SMDC are about 55-fold less cytotoxic than MMAE against the αVβ3-positive WM115 cell line with IC50 values in the nM range, while the cRADfK SMDC is 150-fold less cytotoxic than MMAE. Hence, integrin binding also influences the antiproliferative activity giving a targeting index of 2.8.

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Section: Resultsmentioning

confidence: 99%

Synthesis and Evaluation of a Non-Peptide Small-Molecule Drug Conjugate Targeting Integrin αVβ3

Paulus

Sewald

2022

Front. Chem.

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“…The differences in integrin expression between these cells could play a role in the differences in binding, as integrins might have their unique optimal special presentation. 49 Additionally, phase-separated vesicles containing DSPE-RGD did not bind stronger than uniform DSPE-RGD vesicles in either cell type. FRET analysis to confirm lipid domains demonstrated that DSPE-RGD phase-separated vesicles did exhibit domain formation (Figure S5).…”

Section: Resultsmentioning

confidence: 91%

“…We created uniform and phase-separated vesicles with 10 mol% DOPE-RGD, confirmed phase separation using FRET (Supporting Figure S5), and tested binding to Jurkat and K562 cells, cell lines that both highly express the RGD-binding integrin α 5 β 1 . 49 In uniform vesicles, 10 mol% RGD vesicles exhibited minimal binding on both Jurkat and K562 cells compared to unfunctionalized control vesicles (Figure 4b-e). When RGD was concentrated into lipid domains, binding was greatly enhanced to Jurkats (~15 fold) and K562s (~5 fold) (Figure 4b-e).…”

Section: Phase Separation Can Enhance Binding Of Vesicles With a Ther...mentioning

confidence: 93%

“…In particular, uniform vesicles containing DSPE-RGD bound significantly stronger to Jurkat cells compared to uniform DOPE-RGD vesicles (Figure 4b, c), while in K562 cells this effect was less pronounced (p=0.0503, Figure 4d, e). The differences in integrin expression between these cells could play a role in the differences in binding, as integrins might have their unique optimal special presentation.. 49 Additionally, phase-separated vesicles containing DSPE-RGD did not bind stronger than uniform DSPE-RGD vesicles in either cell type. FRET analysis to confirm lipid domains demonstrated that DSPE-RGD phaseseparated vesicles did exhibit domain formation (Figure S5).…”

Section: The Lipid Saturation Of Rgd Conjugation Affects Cell Bindingmentioning

confidence: 99%

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Enhancing functionalized liposome avidity to cells via lipid phase separation

Sant'Anna

Kamat

2022

Preprint

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The addition of both cell-targeting moieties and polyethylene glycol (PEG) to nanoparticle (NP) drug delivery systems is a standard approach to improve the biodistribution, specificity, and uptake of therapeutic cargo. The spatial presentation of these molecules affects avidity of the NP to target cells in part through an interplay between the local ligand concentration and the steric hindrance imposed by PEG molecules. Here, we show that lipid phase separation in nanoparticles can modulate liposome avidity by changing the proximity of PEG and targeting protein molecules on a nanoparticle surface. Using lipid-anchored nickelnitrilotriacetic acid (Ni-NTA) as a model ligand, we demonstrate that the attachment of lipid anchored Ni-NTA and PEG molecules to distinct lipid domains in nanoparticles can enhance liposome binding to cancer cells by increasing ligand clustering and reducing steric hindrance. We then use this technique to enhance the binding of RGD-modified liposomes, which can bind to integrins overexpressed on many cancer cells. These results demonstrate the potential of lipid phase separation to modulate the spatial presentation of targeting and shielding molecules on nanocarriers, offering a powerful tool to enhance the efficacy of NP drug delivery systems.

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“…Fibronectin concentration is vital to affect the migratory characteristics of the Jurkat cells. CD49e/CD29 (α5β1, VLA-5), which recognizes the RGD-containing fibronectin binding site, and CD49d/CD29 (α4β1, VLA-4), which binds the CS-1 region on fibronectin [ 23 ], control the cells’ attachment and migration movements according to the leukocyte activation status [ 24 , 25 , 26 ]. In this case, we applied a relatively high concentration of fibronectin coating in microchannels for 1 h, and repeated the flash every 10 min with fresh fibronectin solution to achieve the required surface coating density in the channels.…”

Section: Notesmentioning

confidence: 99%

T Cells Chemotaxis Migration Studies with a Multi-Channel Microfluidic Device

Liu

Ren

et al. 2022

Micromachines

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Immune surveillance is dependent on lymphocyte migration and targeted recruitment. This can involve different modes of cell motility ranging from random walk to highly directional environment-guided migration driven by chemotaxis. This study protocol describes a flow-based microfluidic device to perform quantitative multiplex cell migration assays with the potential to investigate in real time the migratory response of T cells at the population or single-cell level. The device also allows for subsequent in situ fixation and direct fluorescence analysis of the cells in the microchannel.

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Selective Integrin Ligands Promote Cell Internalization of the Antineoplastic Agent Fluorouracil

Cited by 7 publications

References 65 publications

Synthesis and Evaluation of a Non-Peptide Small-Molecule Drug Conjugate Targeting Integrin αVβ3

Synthesis and Evaluation of a Non-Peptide Small-Molecule Drug Conjugate Targeting Integrin αVβ3

Enhancing functionalized liposome avidity to cells via lipid phase separation

T Cells Chemotaxis Migration Studies with a Multi-Channel Microfluidic Device

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