Structural characterization and in vivo evaluation of β-hairpin peptidomimetics as specific CXCR4 imaging agents

Wojciech Lesniak, Emilia Sikorska, Hassan Shallal, Babak Behnam Azad, Ala Lisok, Mrudula Pullambhatla, Martin Gilbert Pomper, Sridhar Nimmagadda

Research output: Contribution to journalArticle

Abstract

(Figure Presented) The CXCR4 chemokine receptor is integral to several biological functions and plays a pivotal role in the pathophysiology of many diseases. As such, CXCR4 is an enticing target for the development of imaging and therapeutic agents. Here we report the evaluation of the POL3026 peptidomimetic template for the development of imaging agents that target CXCR4. Structural and conformational analyses of POL3026 and two of its conjugates, DOTA (POL-D) and PEG12-DOTA (POL-PD), by circular dichroism, two-dimensional NMR spectroscopy and molecular dynamics calculations are reported. In silico observations were experimentally verified with in vitro affinity assays and rationalized using crystal structure-based molecular modeling studies. [111In]-labeled DOTA conjugates were assessed in vivo for target specificity in CXCR4 expressing subcutaneous U87 tumors (U87-stb-CXCR4) through single photon emission computed tomography (SPECT/CT) imaging and biodistribution studies. In silico and in vitro studies show that POL3026 and its conjugates demonstrate similar interactions with different micelles that mimic cellular membrane and that the ε-NH2 of lysine7 is critical to maintain high affinity to CXCR4. Modification of this group with DOTA or PEG12-DOTA led to the decrease of IC50 value from 0.087 nM for POL3026 to 0.47 nM and 1.42 nM for POL-D and POL-PD, respectively. In spite of the decreased affinity toward CXCR4, [111In]POL-D and [111In]POL-PD demonstrated high and significant uptake in U87-stb-CXCR4 tumors compared to the control U87 tumors at 90 min and 24 h post injection. Uptake in U87-stb-CXCR4 tumors could be blocked by unlabeled POL3026, indicating specificity of the agents in vivo. These results suggest POL3026 as a promising template to develop new imaging agents that target CXCR4.

Original languageEnglish (US)
Pages (from-to)941-953
Number of pages13
JournalMolecular Pharmaceutics
Volume12
Issue number3
DOIs
StatePublished - Mar 2 2015

Fingerprint

Peptidomimetics
Computer Simulation
Neoplasms
CXCR4 Receptors
Chemokine Receptors
Micelles
Molecular Dynamics Simulation
Circular Dichroism
Single-Photon Emission-Computed Tomography
Molecular Structure
Inhibitory Concentration 50
Magnetic Resonance Spectroscopy
Injections
Membranes
In Vitro Techniques

Keywords

  • chemokine
  • chemokine receptor
  • CXCR4
  • molecular imaging
  • SPECT/CT

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Molecular Medicine
  • Drug Discovery

Cite this

Structural characterization and in vivo evaluation of β-hairpin peptidomimetics as specific CXCR4 imaging agents. / Lesniak, Wojciech; Sikorska, Emilia; Shallal, Hassan; Behnam Azad, Babak; Lisok, Ala; Pullambhatla, Mrudula; Pomper, Martin Gilbert; Nimmagadda, Sridhar.

In: Molecular Pharmaceutics, Vol. 12, No. 3, 02.03.2015, p. 941-953.

Research output: Contribution to journalArticle

@article{690e98f348444efba94a1f4993f32e06,
title = "Structural characterization and in vivo evaluation of β-hairpin peptidomimetics as specific CXCR4 imaging agents",
abstract = "(Figure Presented) The CXCR4 chemokine receptor is integral to several biological functions and plays a pivotal role in the pathophysiology of many diseases. As such, CXCR4 is an enticing target for the development of imaging and therapeutic agents. Here we report the evaluation of the POL3026 peptidomimetic template for the development of imaging agents that target CXCR4. Structural and conformational analyses of POL3026 and two of its conjugates, DOTA (POL-D) and PEG12-DOTA (POL-PD), by circular dichroism, two-dimensional NMR spectroscopy and molecular dynamics calculations are reported. In silico observations were experimentally verified with in vitro affinity assays and rationalized using crystal structure-based molecular modeling studies. [111In]-labeled DOTA conjugates were assessed in vivo for target specificity in CXCR4 expressing subcutaneous U87 tumors (U87-stb-CXCR4) through single photon emission computed tomography (SPECT/CT) imaging and biodistribution studies. In silico and in vitro studies show that POL3026 and its conjugates demonstrate similar interactions with different micelles that mimic cellular membrane and that the ε-NH2 of lysine7 is critical to maintain high affinity to CXCR4. Modification of this group with DOTA or PEG12-DOTA led to the decrease of IC50 value from 0.087 nM for POL3026 to 0.47 nM and 1.42 nM for POL-D and POL-PD, respectively. In spite of the decreased affinity toward CXCR4, [111In]POL-D and [111In]POL-PD demonstrated high and significant uptake in U87-stb-CXCR4 tumors compared to the control U87 tumors at 90 min and 24 h post injection. Uptake in U87-stb-CXCR4 tumors could be blocked by unlabeled POL3026, indicating specificity of the agents in vivo. These results suggest POL3026 as a promising template to develop new imaging agents that target CXCR4.",
keywords = "chemokine, chemokine receptor, CXCR4, molecular imaging, SPECT/CT",
author = "Wojciech Lesniak and Emilia Sikorska and Hassan Shallal and {Behnam Azad}, Babak and Ala Lisok and Mrudula Pullambhatla and Pomper, {Martin Gilbert} and Sridhar Nimmagadda",
year = "2015",
month = "3",
day = "2",
doi = "10.1021/mp500799q",
language = "English (US)",
volume = "12",
pages = "941--953",
journal = "Molecular Pharmaceutics",
issn = "1543-8384",
publisher = "American Chemical Society",
number = "3",

}

TY - JOUR

T1 - Structural characterization and in vivo evaluation of β-hairpin peptidomimetics as specific CXCR4 imaging agents

AU - Lesniak, Wojciech

AU - Sikorska, Emilia

AU - Shallal, Hassan

AU - Behnam Azad, Babak

AU - Lisok, Ala

AU - Pullambhatla, Mrudula

AU - Pomper, Martin Gilbert

AU - Nimmagadda, Sridhar

PY - 2015/3/2

Y1 - 2015/3/2

N2 - (Figure Presented) The CXCR4 chemokine receptor is integral to several biological functions and plays a pivotal role in the pathophysiology of many diseases. As such, CXCR4 is an enticing target for the development of imaging and therapeutic agents. Here we report the evaluation of the POL3026 peptidomimetic template for the development of imaging agents that target CXCR4. Structural and conformational analyses of POL3026 and two of its conjugates, DOTA (POL-D) and PEG12-DOTA (POL-PD), by circular dichroism, two-dimensional NMR spectroscopy and molecular dynamics calculations are reported. In silico observations were experimentally verified with in vitro affinity assays and rationalized using crystal structure-based molecular modeling studies. [111In]-labeled DOTA conjugates were assessed in vivo for target specificity in CXCR4 expressing subcutaneous U87 tumors (U87-stb-CXCR4) through single photon emission computed tomography (SPECT/CT) imaging and biodistribution studies. In silico and in vitro studies show that POL3026 and its conjugates demonstrate similar interactions with different micelles that mimic cellular membrane and that the ε-NH2 of lysine7 is critical to maintain high affinity to CXCR4. Modification of this group with DOTA or PEG12-DOTA led to the decrease of IC50 value from 0.087 nM for POL3026 to 0.47 nM and 1.42 nM for POL-D and POL-PD, respectively. In spite of the decreased affinity toward CXCR4, [111In]POL-D and [111In]POL-PD demonstrated high and significant uptake in U87-stb-CXCR4 tumors compared to the control U87 tumors at 90 min and 24 h post injection. Uptake in U87-stb-CXCR4 tumors could be blocked by unlabeled POL3026, indicating specificity of the agents in vivo. These results suggest POL3026 as a promising template to develop new imaging agents that target CXCR4.

AB - (Figure Presented) The CXCR4 chemokine receptor is integral to several biological functions and plays a pivotal role in the pathophysiology of many diseases. As such, CXCR4 is an enticing target for the development of imaging and therapeutic agents. Here we report the evaluation of the POL3026 peptidomimetic template for the development of imaging agents that target CXCR4. Structural and conformational analyses of POL3026 and two of its conjugates, DOTA (POL-D) and PEG12-DOTA (POL-PD), by circular dichroism, two-dimensional NMR spectroscopy and molecular dynamics calculations are reported. In silico observations were experimentally verified with in vitro affinity assays and rationalized using crystal structure-based molecular modeling studies. [111In]-labeled DOTA conjugates were assessed in vivo for target specificity in CXCR4 expressing subcutaneous U87 tumors (U87-stb-CXCR4) through single photon emission computed tomography (SPECT/CT) imaging and biodistribution studies. In silico and in vitro studies show that POL3026 and its conjugates demonstrate similar interactions with different micelles that mimic cellular membrane and that the ε-NH2 of lysine7 is critical to maintain high affinity to CXCR4. Modification of this group with DOTA or PEG12-DOTA led to the decrease of IC50 value from 0.087 nM for POL3026 to 0.47 nM and 1.42 nM for POL-D and POL-PD, respectively. In spite of the decreased affinity toward CXCR4, [111In]POL-D and [111In]POL-PD demonstrated high and significant uptake in U87-stb-CXCR4 tumors compared to the control U87 tumors at 90 min and 24 h post injection. Uptake in U87-stb-CXCR4 tumors could be blocked by unlabeled POL3026, indicating specificity of the agents in vivo. These results suggest POL3026 as a promising template to develop new imaging agents that target CXCR4.

KW - chemokine

KW - chemokine receptor

KW - CXCR4

KW - molecular imaging

KW - SPECT/CT

UR - http://www.scopus.com/inward/record.url?scp=84923902142&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84923902142&partnerID=8YFLogxK

U2 - 10.1021/mp500799q

DO - 10.1021/mp500799q

M3 - Article

C2 - 25590535

AN - SCOPUS:84923902142

VL - 12

SP - 941

EP - 953

JO - Molecular Pharmaceutics

JF - Molecular Pharmaceutics

SN - 1543-8384

IS - 3

ER -