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Novel Non-Congeneric Derivatives of the Choline Kinase Alpha Inhibitor ICL-CCIC-0019

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Novel Non-Congeneric Derivatives of the Choline Kinase Alpha Inhibitor ICL-CCIC-0019 pharmaceutics Article Novel Non-Congeneric Derivatives of the Choline Kinase Alpha Inhibitor ICL-CCIC-0019 Ning Wang 1, Diana Brickute 1, Marta Braga 1 , Chris Barnes 1, Haonan Lu 1 , Louis Allott 1,2,* and Eric O. Aboagye 1,* 1 Comprehensive Cancer Imaging Centre, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Hammersmith Hospital, London W12 0NN, UK; [email protected] (N.W.); [email protected] (D.B.); [email protected] (M.B.); [email protected] (C.B.); [email protected] (H.L.) 2 Positron Emission Tomography Research Centre, Faculty of Health Sciences, University of Hull, Kingston upon Hull HU6 7RX, UK * Correspondence: [email protected] (L.A.); [email protected] (E.O.A.) Abstract: Choline kinase alpha (CHKA) is a promising target for the development of cancer therapeu- tics. We have previously reported ICL-CCIC-0019, a potent CHKA inhibitor with high cellular activity but with some unfavorable pharmacological properties. In this work, we present an active analogue of ICL-CCIC-0019 bearing a piperazine handle (CK146) to facilitate further structural elaboration of the pharmacophore and thus improve the biological profile. Two different strategies were evaluated in this study: (1) a prodrug approach whereby selective CHKA inhibition could be achieved through modulating the activity of CK146, via the incorporation of an "-(Ac) Lys motif, cleavable by elevated levels of histone deacetylase (HDAC) and cathepsin L (CTSL) in tumour cells; (2) a prostate-specific Citation: Wang, N.; Brickute, D.; membrane antigen (PSMA) receptor targeted delivery strategy. Prodrug (CK145) and PSMA-targeted Braga, M.; Barnes, C.; Lu, H.; Allott, (CK147) derivatives were successfully synthesized and evaluated in vitro. While the exploitation L.; Aboagye, E.O. Novel Non-Congeneric Derivatives of the of CK146 in those two strategies did not deliver the expected results, important and informative Choline Kinase Alpha Inhibitor structure-activity relationships were observed and have been reported. ICL-CCIC-0019. Pharmaceutics 2021, 13, 1078. https://doi.org/10.3390/ Keywords: choline kinase alpha (CHKA) inhibitor; ICL-CCIC-0019; prodrug; PSMA; targeted drug pharmaceutics13071078 delivery; PIK4CB Academic Editor: Luisa Carlota Lopez-Cara 1. Introduction Received: 22 June 2021 Abnormal lipid metabolism is a common feature observed in many cancers [1]. En- Accepted: 9 July 2021 hanced de novo synthesis of lipids in tumorigenesis supports the rapid proliferation of Published: 14 July 2021 cancer cells, cell signaling and tumor survival [2]. The cholinic phenotype, characterized by the overexpression of choline kinase alpha isoform (CHKA) and increased phosphocholine Publisher’s Note: MDPI stays neutral (PCho) levels, is one of the aberrant lipid metabolism pathways revealed in cancer [3]. with regard to jurisdictional claims in Choline kinase is a class of enzymes responsible for generating PCho which is involved in published maps and institutional affil- the biosynthesis of phosphatidylcholine, a major lipid component of cell membranes [4]. iations. Choline kinase exists in at least three isoforms, CHKA1, CHKA2 and CHKB [4], of which the A, but not the B isoform, has been implicated in cancer development [3,5]. Overexpression of CHKA has been reported in a wide range of solid tumors [3,5–12], and correlates with advanced histological tumor grade, poor prognosis and reduced survival rates in breast Copyright: © 2021 by the authors. and non-small-cell lung cancers [9,13]. In prostate cancer (PCa), CHKA is a co-chaperone Licensee MDPI, Basel, Switzerland. for the androgen receptor (AR), thus maintains AR signaling [14]. Taken together, CHKA This article is an open access article is a prognostic marker and a potential therapeutic target for cancer treatment. distributed under the terms and A number of small-molecule CHKA inhibitors have been developed as novel anti- conditions of the Creative Commons Attribution (CC BY) license (https:// cancer strategies for CHKA inhibition [15]. Hemicholinium-3 (HC-3) was the first CHKA creativecommons.org/licenses/by/ inhibitor built around a bis-oxazonium pharmacophore (Figure1)[ 16–18], but exhibited 4.0/). off-target effects on choline transporters, acetyltransferase and acetylcholinesterase [19]. Pharmaceutics 2021, 13, 1078. https://doi.org/10.3390/pharmaceutics13071078 https://www.mdpi.com/journal/pharmaceutics Pharmaceutics 2021, 13, x FOR PEER REVIEW 2 of 17 Pharmaceutics 2021, 13, 1078 2 of 17 off-target effects on choline transporters, acetyltransferase and acetylcholinesterase [19]. MN58BMN58B waswas subsequently subsequently designed designed by by structural structural modifications modifications of HC-3 of HC-3 and demonstratedand demon- goodstrated antiproliferative good antiproliferative potencies potenciesin vitro and in vitroin vivo and(Figure in vivo1)[ (Figure20–22]. The1) [20–22]. replacement The re- of pyridiniumsplacement of inpyridiniums MN58B by in quinoliniums MN58B by quinolin as cationiciums head as cationic groups head elicited groups the bestelicited results the inbest vitro resultsand in vivovitro, and in gave vivo, rise and to RSM-932Agave rise to (Figure RSM-932A1), the (Figure first CHKA 1), the inhibitorfirst CHKA to enterinhibitor Phase to Ienter clinical Phase trials I clinical [23,24 ].trials [23,24]. FigureFigure 1. 1.Chemical Chemical structuresstructures andand pharmacologicalpharmacological datadata of correspondingcorresponding CHKA inhibitors: HC-3 [18,25], [18,25], MN58B MN58B [20,21], [20,21], RSM-932ARSM-932A [ 23[23],], ICL-CCIC-0019 ICL-CCIC-0019 [[26,27]26,27] andand CK14CK14 [[26].26]. InIn recentrecent years, the the x-ray x-ray crystal crystal structur structuree of of CHKA, CHKA, together together with with computational computational in insilico silico techniques techniques have have boosted boosted the the developmen developmentt of CHKA of CHKA inhibitors inhibitors (the (the crystal crystal structure struc- tureof human of human CHKA, CHKA, pdb code: pdb 2CKO) code: 2CKO)[18,20,28–36]. [18,20 ,A28 –series36]. of A symmetrical series of symmetrical N,N-dimethyl-N,N- dimethylaminopyridineaminopyridine (DMAP) (DMAP) derivatives derivatives bearing bearing alkyl linkers alkyl linkers of varying of varying lengths lengths were were de- designedsigned by by Trousil Trousil et et al. al. as as CHKA CHKA inhibitors inhibitors (Figure (Figure 1),1), where DMAP mimicked cationiccationic cholinecholine moietiesmoieties [[26];26]; fromfrom thisthis study, ICL-CCIC-0019ICL-CCIC-0019 was selected for further biological evaluationsevaluations and displayed displayed great great potential potential in in preclinical preclinical studies studies [27]. [27 ICL-CCIC-0019]. ICL-CCIC-0019 selec- se- lectivelytively inhibited inhibited CHKA CHKA activity activity with with potent potent antiproliferative antiproliferative activities activities against against a a broad spectrumspectrum of human cancer cancer cell cell lines lines (IC (IC50:50 0.27: 0.27 ± 0.06± 0.06 μM;µ meanM; mean GI50 of GI 850 cancerof 8 cancer cell lines: cell lines:1.09 μ 1.09M, range:µM, range: 0.38–2.70 0.38–2.70 μM; GIµ50M; range GI50 ofrange 2 normal of 2 normal cell lines: cell 30–120 lines: 30–120μM); ICL-CCIC-µM); ICL- CCIC-00190019 caused caused rapid and rapid sustained and sustained lipid synthesi lipid synthesiss inhibition inhibition by acting by actingon CDP-choline on CDP-choline path- pathwayway and andalso alsoshowed showed potent potent in vivoin vivo anti-tanti-tumorumor activity. activity. The Thecompound, compound, however, however, in- inducedduced mitochondrial mitochondrial changes changes and and slight slight reduct reductionion in inbody body weight weight (not (not below below the the stand- stan- dardard 15–20%) 15–20%) observed observed in in the the rodent rodent model model bear bearinging HCT116 HCT116 xenografts; xenografts; implying implying adverse adverse pharmacologicalpharmacological effectseffects which which precluded precluded clinical clinical translation translation [25 [25].]. Modulating Modulating the the unfavor- unfa- ablevorable properties properties of ICL-CCIC-0019 of ICL-CCIC-0019 was was likely likely to be to challengingbe challenging for for two two reasons: reasons: firstly, firstly, the intrinsicthe intrinsic toxicity toxicity of quaternary of quaternary ammonium ammonium CHKA CHKA inhibitors inhibitors is difficult is difficult to overcome to overcome since theirsince structurestheir structures mimic mimic choline choline and the and inhibition the inhibition of choline of transporterscholine transporters was observed was ob- for theseserved molecules for these [molecules26]. Secondly, [26]. the Secondly, limited th structurale limited diversity structural of diversity ICL-CCIC-0019 of ICL-CCIC- makes direct0019 makes modification direct modification of the molecule of the less molecule feasible. less While feasible. the quaternary While the ammonium quaternary moieties ammo- arenium essential moieties to are the activityessential and to the could activity not be and modified, could not we be hypothesized modified, we that hypothesized controlling thethat release controlling or delivery the release of potent or delivery CHKA inhibitors of potent inCHKA target inhibitors tissue may in reduce target off-target tissue may ac- cumulationreduce off-target and toxicity. accumulation These strategies and toxicity. have Th beenese strategies exemplified have for been a range exemplified
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