Time dependent HPLC analysis of the product ratio of enzymatically reduced prodrug CB1954 by a modified and immobilised nitroreductase

Ball, Patrick; Thompson, Emma; Anderson, Simon; Gwenin, Christopher; Gwenin, Chris

doi:10.1016/j.ejps.2018.11.001

Cited by 10 publications

(34 citation statements)

References 36 publications

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“…NfnB The comparison of the NfnB-Cys:CPP conjugates with our previously reported free NfnB-Cys enzyme kinetics [29] and indicated an alteration in the enzyme's kinetic behaviour, for both NfnB-Cys:CPP conjugates. The k cat and K m values presented are the apparent (app) values measured at 400 µM NADH.…”

Section: Conjugatementioning

confidence: 54%

“…Previously we have shown that after a 30 min reaction time, NfnB-Cys reduced CB1954 to two products (namely 2-NHOH and 4-NHOH) at a ratio of 32:68 respectively [29]. Following the same procedure, conjugates of NfnB-Cys and the CPPs at a 1:1 molar ratio were tested for their ability to reduce CB1954.…”

Section: Hplc Profiles Of Nfnb-cys and Nfnb-cyse:cpp Reaction Productsmentioning

confidence: 99%

“…Following the same procedure, conjugates of NfnB-Cys and the CPPs at a 1:1 molar ratio were tested for their ability to reduce CB1954. Reaction product ratios were established, with the 4-NHOH product eluting at approximately 5 min and the 2-NHOH eluting at 10 min [29,39]. The product ratios obtained for the reactions between NfnB-Cys:CPP conjugates and CB1954 are presented in Table 2; the NfnB-Cys:HR9 produced products at a ratio of 36:64 (2'-NHOH vs 4'-NHOH), whilst the NfnB-Cys:Pep-1 produces the 2-NHOH and 4-NHOH at a ratio of 35:65.…”

Section: Hplc Profiles Of Nfnb-cys and Nfnb-cyse:cpp Reaction Productsmentioning

confidence: 99%

“…We have investigated a novel form of DEPT that involves the incorporation of gold-coated superparamagnetic iron oxide nanoparticles (AuMNP) as the delivery vehicle used to direct the enzyme to the tumour site, an approach termed magnetic nanoparticle directed enzyme prodrug therapy (MNDEPT) [14,27]. A genetically modified nitroreductase (NfnB-Cys) is conjugated to the surface of the nanoparticles [14,28] with conjugates already proven to retain their prodrug reducing capability [29].…”

Section: Introductionmentioning

confidence: 99%

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Cell-Penetrating Peptides as a Tool for the Cellular Uptake of a Genetically Modified Nitroreductase for use in Directed Enzyme Prodrug Therapy

Anderson

Hobbs

Ball

et al. 2019

JFB

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Directed enzyme prodrug therapy (DEPT) involves the delivery of a prodrug-activating enzyme to a solid tumour site, followed by the subsequent activation of an administered prodrug. One of the most studied enzyme–prodrug combinations is the nitroreductase from Escherichia coli (NfnB) with the prodrug CB1954 [5-(aziridin-1-yl)-2,4-dinitro-benzamide]. One of the major issues faced by DEPT is the ability to successfully internalize the enzyme into the target cells. NfnB has previously been genetically modified to contain cysteine residues (NfnB-Cys) which bind to gold nanoparticles for a novel DEPT therapy called magnetic nanoparticle directed enzyme prodrug therapy (MNDEPT). One cellular internalisation method is the use of cell-penetrating peptides (CPPs), which aid cellular internalization of cargo. Here the cell-penetrating peptides: HR9 and Pep-1 were tested for their ability to conjugate with NfnB-Cys. The conjugates were further tested for their potential use in MNDEPT, as well as conjugating with the delivery vector intended for use in MNDEPT and tested for the vectors capability to penetrate into cells.

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Section: Conjugatementioning

confidence: 54%

Section: Hplc Profiles Of Nfnb-cys and Nfnb-cyse:cpp Reaction Productsmentioning

confidence: 99%

Section: Hplc Profiles Of Nfnb-cys and Nfnb-cyse:cpp Reaction Productsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Cell-Penetrating Peptides as a Tool for the Cellular Uptake of a Genetically Modified Nitroreductase for use in Directed Enzyme Prodrug Therapy

Anderson

Hobbs

Ball

et al. 2019

JFB

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“…In an interesting application, Ball et al used a nitroreductase (NfnB) with an engineered histidine and cysteine tag (NfnB-cys) immobilized on an AuNP to activate the prodrug 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954) for potential application in Directed Enzyme Prodrug Therapy (DEPT) for cancer treatment [117]. The prodrug has two nitro groups at the 2 and 4 positions that can be reduced to NHOH.…”

Section: Enzyme Immobilization On Aunpsmentioning

confidence: 99%

Quantum Dots and Gold Nanoparticles as Scaffolds for Enzymatic Enhancement: Recent Advances and the Influence of Nanoparticle Size

et al. 2020

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Nanoparticle scaffolds can impart multiple benefits onto immobilized enzymes including enhanced stability, activity, and recoverability. The magnitude of these benefits is modulated by features inherent to the scaffold–enzyme conjugate, amongst which the size of the nanoscaffold itself can be critically important. In this review, we highlight the benefits of enzyme immobilization on nanoparticles and the factors affecting these benefits using quantum dots and gold nanoparticles as representative materials due to their maturity. We then review recent literature on the use of these scaffolds for enzyme immobilization and as a means to dissect the underlying mechanisms. Detailed analysis of the literature suggests that there is a “sweet-spot” for scaffold size and the ratio of immobilized enzyme to scaffold, with smaller scaffolds and lower enzyme:scaffold ratios generally providing higher enzymatic activities. We anticipate that ongoing studies of enzyme immobilization onto nanoscale scaffolds will continue to sharpen our understanding of what gives rise to beneficial characteristics and allow for the next important step, namely, that of translation to large-scale processes that exploit these properties.

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Evaluation of two xenobiotic reductases from Pseudomonas putida for their suitability for magnetic nanoparticle‐directed enzyme prodrug therapy as a novel approach to cancer treatment

et al. 2020

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Time dependent HPLC analysis of the product ratio of enzymatically reduced prodrug CB1954 by a modified and immobilised nitroreductase

Abstract: Time dependent HPLC analysis of the product ratio of enzymatically reduced prodrug CB1954 by a modified and immobilised nitroreductase Gwenin,

Cited by 10 publications

References 36 publications

Cell-Penetrating Peptides as a Tool for the Cellular Uptake of a Genetically Modified Nitroreductase for use in Directed Enzyme Prodrug Therapy

Cell-Penetrating Peptides as a Tool for the Cellular Uptake of a Genetically Modified Nitroreductase for use in Directed Enzyme Prodrug Therapy

Quantum Dots and Gold Nanoparticles as Scaffolds for Enzymatic Enhancement: Recent Advances and the Influence of Nanoparticle Size

Evaluation of two xenobiotic reductases from Pseudomonas putida for their suitability for magnetic nanoparticle‐directed enzyme prodrug therapy as a novel approach to cancer treatment

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