Metallodendrimer‐sensitised Cytochrome P450 3A4 Electrochemical Biosensor for TB Drugs

Abstract

A cytochrome P450 3A4 (CYP3A4) based enzymatic biosensor was developed with the incorporation of a first-generation copper polypropyleneimine (CuPPI) metallodendrimer for the detection of anti-tuberculosis (anti-TB) drugs. The development of an electrochemical phenotype biosensor for this purpose is still vital since it aids in the ongoing fight against TB by determining metabolic profile. This allows TB treatment to be tailored on an individual patient basis, minimise adverse drug reactions and improve quality of life in TB patients. This simple biosensor was constructed via physical adsorption of CuPPI onto a gold electrode with subsequent electrostatic attachment of CYP3A4. The biosensor was successful in detecting all four first line anti-TB drugs i. e. isoniazid, ethambutol, pyrazinamide and rifampicin with limits of detection ranging from 0.02244 to 0.1072 nM in 0.1 M phosphate buffer. The developed biosensor was then applied towards "real samples" in the form of spiked synthetic urine and plasma. Calibration curves were carried out in the complex matrices, which were diluted with 0.1 M PB. These yielded good LOD in the range of ultra-low micromolar concentration i. e. 0.165-0.884 μM across all drugs. Recovery studies were also successful when detecting the real tablets in both plasma and urine with results ranging from 91.5 % to 108.5 %.

State of the Art on Developments of (Bio)Sensors and Analytical Methods for Rifamycin Antibiotics Determination

“…This can be explained not only by the fact that electrochemical methods have the advantage of being relatively simple, rapid and cheap, but also through the multitude of materials and procedures that allow for the modification of the electrode surface in order to achieve more sensitive and selective determinations. Comparing the RIF therapeutic peak plasma concentration of 8–24 µg/mL (9.72 × 10 −6 –2.92 × 10 −5 mol/L RIF) [ 159 ] with the linear ranges of the electrochemical methods reported in the last 20 years for RIF quantification ( Table 1 ), it can be observed that, with one exception [ 90 ], all these methods could be applied to RIF blood level monitoring, most of them [ 19 , 25 , 26 , 27 , 28 , 36 , 45 , 50 , 52 , 56 , 65 , 66 , 67 , 83 , 84 , 86 , 88 , 104 , 106 , 111 , 112 , 114 , 115 , 116 , 117 , 120 , 121 ] needing a previous dilution step to bring RIF concentration into the linear range of the method. UV-Vis spectrometric methods present narrower linear ranges and are less sensitive; therefore, according to the data summarized in Table 2 , most RIF determinations based on absorbance measurements may be applied as reported for the assessment of RIF blood concentration.…”

“…The anti-TB treatment very often involves the administration of RIF together with other drugs, like ISN and PYR. Therefore, the selectivity and the sensitivity of analytical methods are compulsory features for simultaneous determination of the drugs in both pharmaceuticals and more complex matrices, like biological samples [ 18 , 56 , 68 , 69 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 ]. RIF bioavailability and, therefore, its therapeutic action can be influenced by its physical or chemical interactions with other molecules (peptides, DNA, drugs, nutrients, etc.).…”

“…RIF and RSV ( Figure 1 ) are hydroquinone derivatives (−OH groups in positions 1 and 4, Figure 1 ) and possess a third phenolic −OH moiety (position 8), which can be chemically or electrochemically oxidized [ 40 ] at various solid bare electrodes (e.g., GCE [ 85 ], CPE [ 80 ] and the disposable PGE [ 84 ]) or modified sensors obtained at various substrates, such as GCE [ 28 , 29 , 32 , 37 , 45 , 52 , 65 , 66 , 67 , 68 , 85 , 90 , 106 , 107 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 121 ], SPE [ 36 , 53 , 115 ], CPE [ 21 , 26 , 80 , 88 , 116 , 117 ], ITO [ 118 ], Ag [ 55 ], Au [ 50 , 86 ] and Pt [ 119 , 120 ]. Most often, the electrodes were employed as sensing devices in voltammetric [ 26 , 27 , 28 , 31 , 36 , …”

“…The biosensor reduction peak at −0.080 V, attributed to monooxygenation reaction occurring within the enzyme, linearly increased with the added drug concentration due to the interaction between the antibiotic and the Fe(III) active site of the enzyme. The biosensor had the highest sensitivity for PYR (3000 A × L/mol) and the lowest for RIF (890 A × L/mol), with the corresponding LOQs of 6.80 × 10 −11 mol/L PYR and 3.25 × 10 −10 mol/L RIF [ 86 ].…”

Novel Cytochrome P450-3A4 Enzymatic Nanobiosensor for Lapatinib (a Breast Cancer Drug) Developed on a Poly(anilino-co-4-aminobenzoic Acid-Green-Synthesised Indium Nanoparticle) Platform

“…The use of nanoparticles (NPs) in biosensors to identify cancer carries the promise of improving the survival rate of patients. Due to their various properties, nanoparticles have been widely used in sensing systems to detect and monitor different diseases and viruses [ 15 , 16 , 17 , 18 , 19 , 20 , 21 ]. Likewise, there are a variety of approaches that can be used to create these nanoparticles.…”