Immunoglobulin Fc-fused, neuropilin-1-specific peptide shows efficient tumor tissue penetration and inhibits tumor growth via anti-angiogenesis

Kim, Ye-Jin; Bae, Jeomil; Shin, Tae Hwan; Kang, Se Hun; Jeong, Minwoo; Han, Yang; Park, Ji-Ho; Kim, Seok-Ki; Kim, Yong‐Sung

doi:10.1016/j.jconrel.2015.08.016

Cited by 33 publications

(62 citation statements)

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“…Antibodies were purified from the culture supernatants using a protein-A agarose chromatography column (GE Healthcare) and were extensively dialysed to achieve a final buffer composition of PBS (pH 6.5). The anti-EGFR mAb cetuximab (Erbitux) was expressed and purified5253. Before cell treatments, antibodies were sterilized by filtration using a cellulose acetate membrane filter (0.22 μm, Corning).…”

Section: Methodsmentioning

confidence: 99%

“…Western blotting was performed using specific antibodies following the standard procedure2253. For quantification of western blotting data, band intensities were quantified using ImageJ software and normalized to values of the loading control.…”

Section: Methodsmentioning

confidence: 99%

“…The endosome-depleted cell lysates were then subjected to IP for 2 h at 4 °C with protein A agarose to pull down the antibodies (TMab4, RT11, TMab4-i, and RT11-i), anti-HA antibody (Covance, MMS-101 P), and subsequent incubation with Protein A/G agarose (Santa Cruz, sc-2003) to pull down HA-KRas G12V proteins, or cRaf RBD -immobilized agarose beads (Merck Millipore, 14–278) to pull down the GTP-bound active Ras proteins. The complexes were subsequently washed with lysis buffer and equal precipitates were analysed by western blotting with β-actin as a loading control5355.…”

Section: Methodsmentioning

confidence: 99%

“…After the 6 d incubation, cells were collected and viable cells were counted by trypan blue staining. The results are presented as the percentage of viable cells relative to that of the PBS-treated control53.…”

Section: Methodsmentioning

confidence: 99%

“…After two washes with PBS, the cells were fixed with 4% paraformaldehyde in PBS for 10 min at 25 °C. After staining the nucleus with Hoechst 33342 in PBS for 5 min at 25 °C, center-focused single z-section images were obtained by confocal microscopy as described above2353.…”

Section: Methodsmentioning

confidence: 99%

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Antibody targeting intracellular oncogenic Ras mutants exerts anti-tumour effects after systemic administration

et al. 2017

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Oncogenic Ras mutants, frequently detected in human cancers, are high-priority anticancer drug targets. However, direct inhibition of oncogenic Ras mutants with small molecules has been extremely challenging. Here we report the development of a human IgG1 format antibody, RT11, which internalizes into the cytosol of living cells and selectively binds to the activated GTP-bound form of various oncogenic Ras mutants to block the interactions with effector proteins, thereby suppressing downstream signalling and exerting anti-proliferative effects in a variety of tumour cells harbouring oncogenic Ras mutants. When systemically administered, an RT11 variant with an additional tumour-associated integrin binding moiety for tumour tissue targeting significantly inhibits the in vivo growth of oncogenic Ras-mutated tumour xenografts in mice, but not wild-type Ras-harbouring tumours. Our results demonstrate the feasibility of developing therapeutic antibodies for direct targeting of cytosolic proteins that are inaccessible using current antibody technology.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Antibody targeting intracellular oncogenic Ras mutants exerts anti-tumour effects after systemic administration

et al. 2017

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Assemblies of Peptide‐Cytotoxin Conjugates for Tumor‐Homing Chemotherapy

Wang

Mao

et al. 2019

Adv Funct Materials

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Peptide-drug conjugates are prodrugs that have the advantages of precise molecular structure and the direct exploitation of tumor-homing, penetration or the cellular uptake abilities of the peptides such as the neuropilin-1 receptor targeting peptide. The prodrugs generally have fast blood clearance due to their low molecular weights and thus are made to self-assemble into nanostructures, preferably nanosized micelles and vesicles for intravenous administration, to slow their renal clearance. However, most peptidyl prodrugs usually form precipitates, irregular nanofibers or gels that are unsuitable for intravenous injection. Herein, a arginine-glycine-aspartic acidlysine (RGDK) peptide and cytotoxin 7-ethyl-10-hydroxycamptothecin (SN38) are used to synthesize the tumor-homing prodrugs (SN38-Peps) and explore their structure-micelle formation relationships. A small library of SN38-Peps is obtained using different structures of peptides, linkers, and drug conjugation sites, and the factors affecting the assembly of SN38-Peps as well as the stability of formed micelles are investigated. An optimized SN38-Pep, (MOM)SN38(20)-CRGDK, is finally obtained which forms stable micelles with a hydrodynamic diameter around 110 nm and a fixed drug loading content as high as 35%. The micelles show a prolonged blood circulation, significantly enhanced tumor accumulation, and therefore improved anticancer activity as compared to the non-targeting prodrug and a clinically used anticancer drug.

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Neuropilin: Handyman and Power Broker in the Tumor Microenvironment

Niland

Eble

2020

Advances in Experimental Medicine and Biology

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Immunoglobulin Fc-fused, neuropilin-1-specific peptide shows efficient tumor tissue penetration and inhibits tumor growth via anti-angiogenesis

Cited by 33 publications

References 50 publications

Antibody targeting intracellular oncogenic Ras mutants exerts anti-tumour effects after systemic administration

Antibody targeting intracellular oncogenic Ras mutants exerts anti-tumour effects after systemic administration

Assemblies of Peptide‐Cytotoxin Conjugates for Tumor‐Homing Chemotherapy

Neuropilin: Handyman and Power Broker in the Tumor Microenvironment

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