Synchronized spectrofluorimetric determination of ponatinib and curcumin as an effective therapeutic combination in laboratory prepared mixtures and human plasma samples

Highlights

• This is the first report on synchronous spectrofluorimetric method for simultaneous determination of ponatinib and curcumin.

• Determination of the analytes in spiked human plasma with high recoveries.

• The proposed method was fully validated in compliance with ICH guidelines.

Abstract

Curcumin is a natural product that is frequently utilized in cancer prevention and treatment. The significant benefit of vegetable-derived nutraceuticals in combination with widespread cytostatic medication such as ponatinib is to reduce toxicity and side effects. In this paper, we focus the study on analytical quantification of ponatinib and curcumin through highly sensitive synchronous spectrofluorometric method. Applying this method at Δλ = 160 nm, each of ponatinib and curcumin could be measured at 303 and 412 nm without interference from each others. The diverse experimental factors impacting the performance of the method were studied and optimized. The method exhibited a reasonable linearity in the ranges of 5.0–60.0 and 10.0–200.0 ng/mL for ponatinib and curcumin, respectively with detection limits of 1.48 and 1.22 ng/mL and quantitation limits of 4.49 and 3.68 ng/mL, respectively. The anticipated method was employed for the assessment and evaluation of the studied drugs in the spiked human plasma samples. The mean % recoveries in plasma samples (n = 6) for each of ponatinib and curcumin were 99.84 ± 1.86 and 100.06 ± 2.72, accordingly. The developed method was validated in conformity with the requirements of International Council of Harmonization (ICH).

Introduction

Leukemia is a blood cell malignant disorder that occurs because of gene-promoting growth mutations. This abnormality triggers the development of immature blood cells in the bone marrow. The four primary forms of leukemia are chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), and acute myeloid leukemia (AML) [1].

CML is triggered by gene aberration because there is DNA interchange between long arms of two different chromosomes 9 and 22, contributing to breakpoint cluster region (BCR-ABL) gene expression [2], [3]. In the chronic stage of CML, a tyrosine kinase inhibitor (TKI)-based therapy such as ponatinib is recommended.

Ponatinib (PON) Fig. 1(A) is a benzanilide substituted drug with a molecular formula of C₂₉H₂₇F₃N₆O. It is an effective, orally active TKI that is widely utilized in the treatment of resistant CML and Philadelphia chromosome positive acute lymphoblastic leukemia. It exerts its action via inhibition of tyrosine kinase activity of ABL and T315I mutant kinases [4]. A carbon–carbon triple bond in its structure was designed to target the T315I point mutation within the kinase domain of BCR-ABL [5]. In the literature survey of PON, few methods have been reported for its quantification such as spectrofluorimetry [6] and HPLC [7], [8], [9], [10], [11].

Despite the advanced treatment choices available for leukemia, unique therapeutic strategies are still required. Chemotherapy has a serious impact on human health, causing medical complications such as heart problems, immune loss and the damage of liver and neurons. Because chemotherapeutic agents affect both abnormal cells and normal cells, it causes many unwanted side effects as loss of hair, appetite besides mouth sores, diarrhea, nausea and vomiting [2].

Curcuminoids, which are natural compounds, are used in a variety of leukemia treatments. Curcumin has gained popularity in recent years, not only as a food additive but also as a vital medicine [2].

Curcumin (CUR) Fig. 1(B) is the active ingredient localized in Curcuma longa rhizome. It has powerful tyrosine kinase activity [12]. It was identified also as a bioactive compound have antibacterial, anti-inflammatory, anti-diabetic, antioxidant, cholesterol lowering activity [13], [14], [15]. Additionally, it has anti-platelet aggregation, anti-carcinogenic and chemo-preventive property [16]. CUR is an impressive compound that can enhance the treatment of CML patients in combination with typical TKI such as ponatinib as it triggered the up-regulation of miR-196b and reduced BCR-ABL at the level of mRNA and protein, resulting in leukemic cell growth inhibition [17].

Not only curcumin is effective in conjunction with chemotherapy but also, it is indicated for chemoprevention [18].

Several methods have been reported in the literature for the determination of CUR such as spectrophotometry [19], [20], [21] spectrofluorimetry [22], [23], electrochemical methods [24], [25], [26], [27], [28], and HPLC [29], [30], [31], [32], [33].

To date, no method for the simultaneous quantification of PON and CUR as co-administered drugs in biological fluids has been documented. This necessitates establishment of a responsive procedure for simultaneous estimation of both drugs for their therapeutic drug monitoring. Applying conventional spectrofluorimetric approach, the normal emission fluorescence spectra of the two studied drugs are strongly overlapped, and therefore this method did not contribute to the resolution of such mixture. The problem of spectral overlapping was solved using constant wavelength synchronous spectrofluorimetric approach. Upon applying this method, both PON and CUR have separated synchronous fluorescence emission peaks with intensities measurable at 303 and 412 nm, respectively. The spectrofluorimetric approach is distinguished by its sensitivity and selectivity, but selectivity concerns can arise owing to the overlap of the broad band spectrum mostly during analysis of multi-component samples. This overlapping problem could be solved by applying synchronous fluorescence spectroscopy (SFS) which has several virtues over conventional fluorescence method including clear spectra, high selectivity and minimal interference [34], [35], [36]. SFS behaves as a very simple and powerful method for quantitative evaluation of drugs studied in a single run within a reasonable time due to its sharp and narrow spectrum [37], [38], [39], [40], [41]. Based on the chemical properties of the studied drugs and their high therapeutic utility in the treatment of myeloid leukemia, the SFS technique was introduced for their synchronized estimation in spiked human plasma for the purpose of their therapeutic drug monitoring.