NGR Tumor-Homing Peptides

Structural Requirements for Effective APN (CD13) Targeting

Alessandra Graziadio, Matteo Zanda, Simona Frau, Ian N. Fleming, Manuele Musolino, Sergio Dall'Angelo, Massimiliano Baldassarre, Monica Piras

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Abstract

Cyclic CNGRC (cCNGRC) peptides are very important targeting ligands for Aminopeptidase N (APN or CD13), which is overexpressed on the surface of many cancer cells. In this work we have (1) developed an efficient solid-phase synthesis and (2) tested on purified porcine APN and APN-expressing human cells two different classes of cCNGRC peptides: the first carrying a biotin affinity tag or a fluorescent tag attached to the carboxyl Arg-Cys-COOH terminus and the second with the tags attached to the amino H2N-Cys-Asn terminus. Carboxyl-terminus functionalised cCNGRC peptides 3, 6, 8 showed good affinity for porcine APN and very good capacity to target and be internalised into APN-expressing cells. In contrast, amino-terminus functionalised cCNGRC peptides 4, 5, 7 displayed significantly decreased affinity and targeting capacity. These results, which are in agreement with the recently reported X-ray structure of a cCNGRC peptide bound to APN showing important stabilising interactions between the unprotected cCNGRC amino terminus and the APN active site, indicate that the carboxyl and not the amino-terminus of cCNGRC peptides should be used as a ‘handle’ for the attachment of toxic payloads for therapy or isotopically labelled functions for imaging and nuclear medicine.
Original languageEnglish
Pages (from-to)1332-1340
Number of pages9
JournalBioconjugate Chemistry
Volume27
Issue number5
Early online date14 Apr 2016
DOIs
Publication statusPublished - 18 May 2016

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Cyclic Peptides
Peptides
Tumors
Neoplasms
Cells
CD13 Antigens
Swine
Nuclear medicine
Poisons
Biotin
Solid-Phase Synthesis Techniques
Nuclear Medicine
Catalytic Domain
Ligands
Imaging techniques
X rays
X-Rays

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NGR Tumor-Homing Peptides : Structural Requirements for Effective APN (CD13) Targeting. / Graziadio, Alessandra; Zanda, Matteo; Frau, Simona; Fleming, Ian N.; Musolino, Manuele; Dall'Angelo, Sergio; Baldassarre, Massimiliano; Piras, Monica.

In: Bioconjugate Chemistry, Vol. 27, No. 5, 18.05.2016, p. 1332-1340.

Research output: Contribution to journalArticle

Graziadio, Alessandra ; Zanda, Matteo ; Frau, Simona ; Fleming, Ian N. ; Musolino, Manuele ; Dall'Angelo, Sergio ; Baldassarre, Massimiliano ; Piras, Monica. / NGR Tumor-Homing Peptides : Structural Requirements for Effective APN (CD13) Targeting. In: Bioconjugate Chemistry. 2016 ; Vol. 27, No. 5. pp. 1332-1340.
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abstract = "Cyclic CNGRC (cCNGRC) peptides are very important targeting ligands for Aminopeptidase N (APN or CD13), which is overexpressed on the surface of many cancer cells. In this work we have (1) developed an efficient solid-phase synthesis and (2) tested on purified porcine APN and APN-expressing human cells two different classes of cCNGRC peptides: the first carrying a biotin affinity tag or a fluorescent tag attached to the carboxyl Arg-Cys-COOH terminus and the second with the tags attached to the amino H2N-Cys-Asn terminus. Carboxyl-terminus functionalised cCNGRC peptides 3, 6, 8 showed good affinity for porcine APN and very good capacity to target and be internalised into APN-expressing cells. In contrast, amino-terminus functionalised cCNGRC peptides 4, 5, 7 displayed significantly decreased affinity and targeting capacity. These results, which are in agreement with the recently reported X-ray structure of a cCNGRC peptide bound to APN showing important stabilising interactions between the unprotected cCNGRC amino terminus and the APN active site, indicate that the carboxyl and not the amino-terminus of cCNGRC peptides should be used as a ‘handle’ for the attachment of toxic payloads for therapy or isotopically labelled functions for imaging and nuclear medicine.",
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AU - Fleming, Ian N.

AU - Musolino, Manuele

AU - Dall'Angelo, Sergio

AU - Baldassarre, Massimiliano

AU - Piras, Monica

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AB - Cyclic CNGRC (cCNGRC) peptides are very important targeting ligands for Aminopeptidase N (APN or CD13), which is overexpressed on the surface of many cancer cells. In this work we have (1) developed an efficient solid-phase synthesis and (2) tested on purified porcine APN and APN-expressing human cells two different classes of cCNGRC peptides: the first carrying a biotin affinity tag or a fluorescent tag attached to the carboxyl Arg-Cys-COOH terminus and the second with the tags attached to the amino H2N-Cys-Asn terminus. Carboxyl-terminus functionalised cCNGRC peptides 3, 6, 8 showed good affinity for porcine APN and very good capacity to target and be internalised into APN-expressing cells. In contrast, amino-terminus functionalised cCNGRC peptides 4, 5, 7 displayed significantly decreased affinity and targeting capacity. These results, which are in agreement with the recently reported X-ray structure of a cCNGRC peptide bound to APN showing important stabilising interactions between the unprotected cCNGRC amino terminus and the APN active site, indicate that the carboxyl and not the amino-terminus of cCNGRC peptides should be used as a ‘handle’ for the attachment of toxic payloads for therapy or isotopically labelled functions for imaging and nuclear medicine.

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