In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor
Gastrin-releasing peptide (GRP) receptors are overexpressed on several types of human cancer cells, including breast, prostate, small cell lung, and pancreatic cancers. Bombesin (BBN), a 14–amino acid peptide that is an analogue of human GRP, binds to GRP receptors with very high affinity and specif...
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2007-05-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.2310/7290.2007.00013 |
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| author | Lixin Ma Ping Yu Bhadrasetty Veerendra Tammy L. Rold Lauren Retzloff Adam Prasanphanich Gary Sieckman Timothy J. Hoffman Wynn A. Volkert Charles J. Smith |
| author_facet | Lixin Ma Ping Yu Bhadrasetty Veerendra Tammy L. Rold Lauren Retzloff Adam Prasanphanich Gary Sieckman Timothy J. Hoffman Wynn A. Volkert Charles J. Smith |
| author_sort | Lixin Ma |
| collection | DOAJ |
| description | Gastrin-releasing peptide (GRP) receptors are overexpressed on several types of human cancer cells, including breast, prostate, small cell lung, and pancreatic cancers. Bombesin (BBN), a 14–amino acid peptide that is an analogue of human GRP, binds to GRP receptors with very high affinity and specificity. The aim of this study was to develop a new fluorescent probe based on BBN having high tumor uptake and optimal pharmacokinetics for specific targeting and optical imaging of human breast cancer tissue. In this study, solid-phase peptide synthesis was used to produce H 2 N-glycylglycylglycine-BBN[7–14]NH 2 peptide with the following general sequence: H 2 N-G-G-G-Q-W-A-V-G-H-L-M-(NH 2 ). This conjugate was purified by reversed-phase high-performance liquid chromatography and characterized by electrospray-ionization mass spectra. The fluorescent probe Alexa Fluor 680-G-G-G-BBN[7–14]NH 2 conjugate was prepared by reaction of Alexa Fluor 680 succinimidyl ester to H 2 N-G-G-G-BBN[7–14]NH 2 in dimethylformamide (DMF). In vitro competitive binding assays, using 125 I-Tyr 4 -BBN as the radiolabeling gold standard, demonstrated an inhibitory concentration 50% value of 7.7 ± 1.4 nM in human T-47D breast cancer cells. Confocal fluorescence microscopy images of Alexa Fluor 680-G-G-G-BBN[7–14]NH 2 in human T-47D breast cancer cells indicated specific uptake, internalization, and receptor blocking of the fluorescent bioprobe in vitro. In vivo investigations in SCID mice bearing xenografted T-47D breast cancer lesions demonstrated the ability of this new conjugate to specifically target tumor tissue with high selectivity and affinity. |
| format | Article |
| id | doaj-art-21f1c725c8754d2bb32daed43a154fe7 |
| institution | Kabale University |
| issn | 1536-0121 |
| language | English |
| publishDate | 2007-05-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj-art-21f1c725c8754d2bb32daed43a154fe72025-01-03T01:24:45ZengSAGE PublishingMolecular Imaging1536-01212007-05-01610.2310/7290.2007.0001310.2310_7290.2007.00013In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide ReceptorLixin MaPing YuBhadrasetty VeerendraTammy L. RoldLauren RetzloffAdam PrasanphanichGary SieckmanTimothy J. HoffmanWynn A. VolkertCharles J. SmithGastrin-releasing peptide (GRP) receptors are overexpressed on several types of human cancer cells, including breast, prostate, small cell lung, and pancreatic cancers. Bombesin (BBN), a 14–amino acid peptide that is an analogue of human GRP, binds to GRP receptors with very high affinity and specificity. The aim of this study was to develop a new fluorescent probe based on BBN having high tumor uptake and optimal pharmacokinetics for specific targeting and optical imaging of human breast cancer tissue. In this study, solid-phase peptide synthesis was used to produce H 2 N-glycylglycylglycine-BBN[7–14]NH 2 peptide with the following general sequence: H 2 N-G-G-G-Q-W-A-V-G-H-L-M-(NH 2 ). This conjugate was purified by reversed-phase high-performance liquid chromatography and characterized by electrospray-ionization mass spectra. The fluorescent probe Alexa Fluor 680-G-G-G-BBN[7–14]NH 2 conjugate was prepared by reaction of Alexa Fluor 680 succinimidyl ester to H 2 N-G-G-G-BBN[7–14]NH 2 in dimethylformamide (DMF). In vitro competitive binding assays, using 125 I-Tyr 4 -BBN as the radiolabeling gold standard, demonstrated an inhibitory concentration 50% value of 7.7 ± 1.4 nM in human T-47D breast cancer cells. Confocal fluorescence microscopy images of Alexa Fluor 680-G-G-G-BBN[7–14]NH 2 in human T-47D breast cancer cells indicated specific uptake, internalization, and receptor blocking of the fluorescent bioprobe in vitro. In vivo investigations in SCID mice bearing xenografted T-47D breast cancer lesions demonstrated the ability of this new conjugate to specifically target tumor tissue with high selectivity and affinity.https://doi.org/10.2310/7290.2007.00013 |
| spellingShingle | Lixin Ma Ping Yu Bhadrasetty Veerendra Tammy L. Rold Lauren Retzloff Adam Prasanphanich Gary Sieckman Timothy J. Hoffman Wynn A. Volkert Charles J. Smith In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor Molecular Imaging |
| title | In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor |
| title_full | In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor |
| title_fullStr | In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor |
| title_full_unstemmed | In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor |
| title_short | In Vitro and In Vivo Evaluation of Alexa Fluor 680-Bombesin[7–14]NH Peptide Conjugate, a High-Affinity Fluorescent Probe with High Selectivity for the Gastrin-Releasing Peptide Receptor |
| title_sort | in vitro and in vivo evaluation of alexa fluor 680 bombesin 7 14 nh peptide conjugate a high affinity fluorescent probe with high selectivity for the gastrin releasing peptide receptor |
| url | https://doi.org/10.2310/7290.2007.00013 |
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