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Article

Anticancer Acridones, Part 2—Acronycine-Type Derivatives Modified with 2,5-Dihydro-1,2,4-Triazine Moiety: Synthesis and In Vitro Evaluation

by
Andrey A. Zonov
1,
Ramil F. Fatykhov
1,
Igor A. Khalymbadzha
1,2,*,
Ainur D. Sharapov
1,3,
Anastasia P. Potapova
1,
Ilya I. Butorin
1,
Vsevolod V. Melekhin
1,
Anastasia V. Paramonova
1 and
Emiliya V. Nosova
1
1
Department of Organic and Biomolecular Chemistry, Ural Federal University, 620002 Yekaterinburg, Russia
2
Research Center for Translational Medicine, Sirius University of Science and Technology, Olympic Ave. 1, 354340 Sochi, Russia
3
Department of Biochemistry, Ural State Medical University, Repina 3, 620028 Yekaterinburg, Russia
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2026, 27(4), 1969; https://doi.org/10.3390/ijms27041969
Submission received: 2 December 2025 / Revised: 31 January 2026 / Accepted: 7 February 2026 / Published: 18 February 2026
(This article belongs to the Special Issue Ongoing Anticancer Agents)
Browse Figures
Review Reports Versions Notes

Abstract

This manuscript presents the synthesis of eight novel noracronycine derivatives containing 1,2,4-triazine moiety and evaluates their anticancer activity in vitro. The obtained compounds exhibit activity in the micromolar range and show selectivity towards glioblastoma A172 and breast cancer Hs578T cells. Compounds incorporating a dihydrotriazine moiety demonstrate an enhanced anticancer profile when compared to a noracronycine derivative lacking a triazine substituent. Furthermore, introducing a pyridyl group into the triazine core increases selective cytotoxicity toward cancerous cells. The lead compound exhibits an IC50 value of 3.4 μM for glioblastoma A172, with a selectivity index of 7.59. Mechanistic studies reveal that the obtained compounds slow down cell division, while no significant apoptosis was detected.

1. Introduction

Cancer is a group of diseases characterized by uncontrolled growth and spread of abnormal cells in the body [1]. Presently, it imposes a considerable strain on the global healthcare infrastructure, accounting for nearly 19 million newly diagnosed malignancies annually, resulting in approximately 10 million fatalities [2]. Among the prevalent forms globally are breast, lung, colorectal, prostate, and skin cancers, jointly representing roughly half of all newly reported cases.
Natural compounds are a privileged class for searching for new antitumor agents [3]. This strategy is a successful one that has yielded many compounds in clinical practice [4,5]. Post-modification of natural leads provides a simpler alternative to total synthesis and can enhance the antitumor activity of the parent compounds. For example, docetaxel, a modified paclitaxel, has a number of advantages over its parent compound. Replacement of the benzoyl group with (tert-butoxy)carbonyl moiety results in broader cell cycle activity and increased affinity for the β-tubulin binding site. In addition, docetaxel demonstrates superior cellular uptake and reduced efflux from tumor cells compared to paclitaxel, thereby prolonging intracellular retention and elevating intratumoral concentrations [6].
The acridone framework is extensively represented among naturally occurring antitumor agents, particularly in compounds displaying wide-ranging activity across diverse cancer cell lines [7,8,9]. Acronycine 1 and its analogs, belonging to the subclass of acridone alkaloids, originate from the Australian scrub ash Baurella simplicifolia (Endl.) Hartley and Acronchia baueri Schott (Rutaceae) [10]. These compounds demonstrate the broadest spectrum of in vivo antitumor activity against numerous solid tumors, including carcinoma, sarcoma, myeloma, and melanoma [11,12]. Despite this promising profile, the low solubility of acronycine does not allow its effective use as an antitumor agent [13].
Modifications to the chemical structure of acronycine have enabled the development of several analogs with significantly increased activity [14,15,16,17,18]. For example, diacetoxylation yields a product (compound 2) whose cytotoxic potency rises by 3–8 times depending on the stereochemistry of the acetoxy groups (Figure 1). Furthermore, formation of a cyclic carbonate (compound 3) further amplifies this effect [14]. In more recent developments, researchers have obtained analogs of acronycine with an expanded dimethylpyran ring (Figure 1). The resulting ring-enlarged compounds 5 and 6 display comparable bioactivity profiles against specific human leukemic cell lines MV4-11 and ML1, similar to the parent compounds [19].
It is known that the antitumor activity of acronycine is associated with cytochrome-mediated epoxidation at the double bond of the pyran cycle, yielding an epoxide. The epoxide 4 (Figure 1), as well as the acetyl derivatives (as shown in Figure 1), covalently binds to the amino group of guanines in the DNA strand, initiating a series of cellular events culminating in cell death [20,21].
Previously, our group modified 1-hydroxy-3-methoxy-acridone 7 by reacting with 1,2,4-triazines to obtain a series of cytotoxic acridone derivatives 8, as reported in our previous study [22] and illustrated in Figure 2. Notably, these compounds lack the double bond that may form epoxide responsible for the anticancer activity of acronycine. Nonetheless, they demonstrate good anticancer activity, with IC50 values of 1.74 to 14.6 μM, significantly superior to that of the parent 1-hydroxy-3-methoxyacridone 7.
In continuation of our previous work, we now report on the synthesis and in vitro antitumor evaluation of a new series of acronycine-like derivatives 9, each featuring a pyran ring containing a double-bond moiety (highlighted in magenta in Figure 2).

2. Results and Discussion

2.1. Synthesis

The initial step comprises the reaction of 1,3-dihydroxyacridone 7a with 1,2,4-triazine, which should give the formation of the desired product 8a (Scheme 1, route A). Although the reaction did produce 8a, it also generated an unidentified impurity, accounting for approximately 50% of the mixture based on the 1H NMR analysis. Unfortunately, attempts to purify product 8a by crystallization were unsuccessful, while chromatographic purification was not effective due to the poor solubility of 8a in typical organic solvents. Consequently, this direction was abandoned.
Since route A did not provide the desired product in pure form, an alternative strategy B was developed. The first step involves annulation of the pyran ring using senecialdehyde and 1,3-dihydroxyacridone 7a, a process that has been extensively described in the prior literature [8,18,23,24]. Initially, following the protocol established by Motiur Rahman [8], we encountered difficulties as the reaction only resulted in recovery of the starting materials. Subsequently, the target angular product 10 was obtained in a satisfactory yield, without the need for chromatographic purification, by adopting the methodology outlined by Lee [23].
Having obtained des-N-methylnoracronycine 10, we tried to apply a previously published procedure for introducing the triazine functionality onto the pyranoacridone scaffold. Specifically, heating compound 10 with 3 equiv. MsOH in acetic acid [22,24] afforded the desired product 10 in only 24% yield (Table 1, entry 1). When BF3·Et2O was used as a catalyst in a methanol solution [25], the starting compounds were isolated from the reaction mixture (Table 1, entry 2). Subjecting compound 10 to heating conditions in trifluoroacetic acid (TFA) led to complete degradation of the starting material, presumably due to cleavage of the pyran ring structure (Table 1, entry 3). The desired product 9a was obtained in good yield solely through adaptation of a recently reported procedure employing a heptafluorobutanoic acid (HFBA)-catalyzed reaction conducted in acetic acid [26,27].
To explore the scope of this transformation, diverse 1,2,4-triazine derivatives were incorporated into the reaction effectively (Scheme 2).
The structures of the obtained compounds were confirmed by NMR spectroscopy. In the 1H NMR spectrum, the resonance peak originally located at 6.0 ppm, attributable to the proton bonded to the C5 carbon atom of the pyranoacridone moiety, disappears entirely. Simultaneously, a characteristic signal at 6.6 ppm, indicative of the presence of a hydrogen atom attached to an sp3-hybridized carbon center within the newly formed dihydrotriazine core, is registered [28,29,30]. The presence of the heptafluorobutyl counterion is confirmed by the carboxyl group signal, which is registered as a triplet with an SSCC of 22 Hz at 158 ppm in the 13C NMR spectrum. The other signals could only be detected in the case of compound 9c, since it dissolves well in DMSO-d6, allowing high concentrations to be achieved (see Supplementary Figure S29). In the IR spectra, the compounds of series 9 exhibit a carbonyl group signal at 1680 cm−1; the carboxyl group (of HFBA anion) may be attributed to a broad signal at 3500 cm−1, which overlaps with the stretching vibrations of the phenolic hydroxyl group; the carbon-fluorine bond produces intense absorption at around 1200 cm−1.

2.2. Anticancer Activity

2.2.1. MTT Assay

The concentration–viability response curves of compounds 9b,d,e,g and 10 on A172, HEK-293, and Hs578T obtained in the MTT assay are provided in Figure 3.
The obtained curves were S-shaped and allowed the IC50 values to be calculated (Figure 4 and Supplementary Table S1).
Based on the results presented in Figure 4 and Supplementary Table S2, compounds 9g and 9d demonstrated greater cytotoxic efficacy against A172 cells relative to HEK-293 cells. Compounds 9b and 9e exhibited greater efficacy in inhibiting the growth of Hs578T cells compared to both A172 and HEK-293 cells. In contrast, compound 10 yielded similar IC50 values against all three cell lines.
Additionally, compounds containing a diaryltriazine ring exhibit increased activity and improved selectivity, in contrast to 9h, the ethylthio-derivative lacking two aryl substituents, which demonstrates equivalent activity to its parent compound 10. Notably, compounds 9f and 9g, featuring a pyridyltriazine substituent, possess especially desirable characteristics.
To exclude the cytotoxic effects of the perfluorinated acid counterion, which has been reported for certain cells [31,32,33], an MTT assay was performed with HFBA at various concentrations. As a result, no cytotoxic effect of HFBA was detected on either A172 glioblastoma cells or HEK-293 human embryonic kidney cells at concentrations of 2 to 256 μM (see Supplementary Figures S33 and S34). This concentration range greatly exceeds the cytotoxic concentrations found in the MTT assay (up to 50 μM). Thus, the cytotoxicity of the obtained samples is associated with the heterocyclic system, and not the acid counterion.
The activity of compound 9g was also studied on colon adenocarcinoma HCT116 and human duodenal adenocarcinoma HuTu 80 cancer cell lines. Compound 9g exhibits moderate cytotoxic activity against these cancer types (IC50 = 37.9374 ± 1.1451 and 18.4788 ± 0.3409 μM for HCT116 and HuTu 80, respectively).

2.2.2. Detection of Apoptosis

To confirm the induction of apoptosis, Annexin V/propidium iodine binding studies were performed. The results showed that very few A172 cells were seen in the early apoptotic stage or as being necrotic after the treatment with 9g as well as cisplatin (Supplementary Table S2, Figure 5).
Interestingly, the total number of cells after the treatment with the compounds was slightly lower than in the untreated group. In comparison with cisplatin treatment, treatment with compound 9g led to a reduction in the percentages of both early and late apoptotic cells (AnV+/PI− and AnV+/PI+, respectively), whereas the percentage of necrotic cells (AnV−/PI+) increased.

2.2.3. Mitochondrial Dysfunction

Mitochondrial membrane potential (ΔΨm) is a key indicator of cell health, and its reduction is a sign of irreversible pathological processes induced by one of the mechanisms of cell death.
The effect of the compounds on the mitochondrial membrane potential (ΔΨm) of A172 cells was determined using JC-1 staining. The results suggest that compound 9g reduced the mitochondrial membrane potential by approximately 50% (Figure 6). It should also be noted that the total number of cells after treatment with the studied compound was lower, and the cells were larger than those in the untreated group.

2.2.4. Proliferation

Fluorescent staining of A172 cells was performed using the red-fluorescent probe iF 555-Tyramide. Before staining, cells were incubated for 24 h with the tested compound 9g and for 2h with 5-ethynyl-2′-deoxyuridine (EdU). The results showed a significant decrease in DNA replication activity (p < 0.001). Moreover, the total number of cells was 1.5 times lower compared to untreated controls, which may indicate non-specific inhibition of cell proliferation. Cisplatin, which was used for comparison as a positive control, almost completely blocked DNA replication (Figure 7).

2.2.5. Mechanism of Anticancer Activity

The compounds under analysis may exert their effects through a combination of chemically induced toxicity mechanisms. Although their exact modes of action remain under investigation, certain acridine derivatives have demonstrated potent inhibition of topoisomerase II in vitro [34,35], as well as telomerase and protein kinases [11]. Additionally, these compounds have been found capable of forming both binary drug/DNA complexes and ternary drug/DNA/topo complexes [36].
Another crucial aspect is the role of acronycine epoxide, a highly cytotoxic agent [37,38,39]. This derivative has been documented to disrupt DNA synthesis and repair processes, or affect cellular signaling pathways such as MAPK, NF-κB, and PI3K/Akt [37].

3. Materials and Methods

3.1. Synthesis

Starting materials were purchased from Merck and used without purification. Compounds 7a and 10 were prepared as previously reported [23,40].
1H and 13C NMR spectra were recorded on a Bruker Avance-400 spectrometer (400 MHz) or Bruker NEO (600 MHz). IR spectra were measured on a LUMOS-Bruker IR–Fourier spectrometer in potassium bromide tablets. Elemental analysis was performed on a PE2400 II PerkinElmer CHN analyzer.
Protocol for synthesis of compounds 9a–h.
Noracronycine 10 [23] (1.0 mmol, 293 mg) and 1,2,4-triazine [25,26,27,28,29,30] (1.0 mmol) were dissolved in acetic acid (10 mL), and heptafluorobutanoic acid (3.0 mmol, 390 μL) was added. The mixture was heated for 15 min, cooled, and the precipitate was filtered off, washed with acetic acid, and dried. Images of 1H, 13C, and 19F NMR spectra and IR spectra are provided in Supplementary Figures S1–S32.
5-(3,6-Diphenyl-2,5-dihydro-1,2,4-triazin-5-yl)-6-hydroxy-3,3-dimethyl-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9a. Yield 60% (444 mg).
1H NMR (600 MHz, DMSO-d6) δ 13.46 (br s, 1H, OH), 11.44 (s, 1H, N12H), 11.26 (br s, 1H, N2′H), 8.18–8.20 (m, 1H, C8H), 7.91–7.92 (m, 2H, HAr), 7.77–7.92 (m, 5H, C10H + HAr), 7.67–7.69 (m, 2H, HAr), 7.43–7.45 (m, 3H, HAr), 7.33–7.36 (m, 1H, HAr), 7.07 (d, J = 10.1 Hz, 1H, C1H), 6.71 (s, 1H, C5′H), 5.72 (d, J = 10.1 Hz, 1H, C2H), 1.39 (s, 3H, C3Me), 1.37 (s, 3H, C3Me).
13C NMR (151 MHz, DMSO-d6) δ 180.97 (C=O), 161.28 (C6), 157.78 (C4a), 157.48 (t, 2JC-F = 22.1 Hz, CF2-CO2H), 152.54 (C-Ph), 149.69 (C-Ph’), 140.91 (C11a), 138.04 (C12a), 134.59 (C10), 134.36 (i-CPh), 132.66 (p-CAr), 131.40 (p-CPh), 129.42 (m-CPh, 2C), 128.77 (m-CAr, 2C), 128.11 (o-CAr 2C), 125.87 (o-CAr, 2C), 125.67 (C8), 124.80 (C2 + i-CPh), 122.55 (C9), 118.61 (C7a), 117.87 (C11), 115.51 (C1), 105.57 (C5), 102.93 (C6a), 97.95 (C12b), 78.71 (C3), 41.77 (C5′), 27.93 (C3Me), 27.18 (C3Me), C3F7- group is not registered due to extensive spin–spin coupling.
19F NMR (565 MHz, DMSO-d6) δ −80.11 (t, J = 8.2 Hz), –115.76 (q, J = 8.2 Hz), −126.17.
IR: 1684 (C=O), 1226 (C–F), 1204 (C–F), 1148 (C–F) cm−1.
Calculated, %: C, 60.00; H, 3.67; N, 7.56. Founded, %: C, 60.16; H, 3.65; N, 7.38.
6-Hydroxy-3,3-dimethyl-5-(3-phenyl-6-(o-tolyl)-2,5-dihydro-1,2,4-triazin-5-yl)-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9b. Yield 59% (445 mg).
1H NMR (400 MHz, DMSO-d6) δ 13.43 (br s, 1H, OH), 11.47 (s, 1H, N12H), 11.23 (br s, 1H, N2′H), 8.18–8.20 (m, 1H, C8H), 7.75–7.83 (m, 7H, C10H + HAr), 7.65–7.69 (m, 2H, HAr), 7.32–7.35 (m, 1H, HAr), 7.23 (d, J = 8.0 Hz, 2H, HTol), 7.08 (d, J = 10.1 Hz, 1H, C1H), 6.68 (s, 1H, C5′H), 5.71 (d, J = 10.1 Hz, 1H, C2H), 2.25 (s, 3H, MeTol), 1.39 (s, 3H, C3Me), 1.35 (s, 3H, C3Me).
13C NMR (101 MHz, DMSO-d6) δ 180.97 (C=O), 161.19 (C6), 157.79 (C4a), 157.57 (t, 2JC-F = 22.1 Hz, CF2-CO2H), 152.45 (C-Ar), 149.72 (C-Ar’), 141.52, 140.92 (C11a), 138.02 (C12a), 134.57 (C10), 134.30, 129.90, 129.41 (2C), 129.35 (2C), 128.10 (2C), 125.83 (2C), 125.67 (C8), 124.89, 124.80 (C2), 122.53 (C9), 118.61 (C7a), 117.88 (C11), 115.53 (C1), 105.70 (C5), 102.94 (C6a), 97.97 (C12b), 78.68 (C3), 41.73 (C5′), 27.97 (C3Me), 27.15 (C3Me), 20.92 (MeTol), C3F7- group is not registered due to extensive C-F spin–spin coupling.
19F NMR (565 MHz, DMSO-d6) δ −80.10 (t, J = 8.7 Hz), −115.70 (q, J = 8.7 Hz), −126.15.
IR: 1683 (C=O), 1204 (C–F), 1146 (C–F) cm−1.
Calculated, %: C, 60.48; H, 3.87; N, 7.42. Founded, %: C, 60.61; H, 3.80; N, 7.37.
6-Hydroxy-5-(6-(4-methoxyphenyl)-3-phenyl-2,5-dihydro-1,2,4-triazin-5-yl)-3,3-dimethyl-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9c. Yield 55% (423 mg).
1H NMR (600 MHz, DMSO-d6) δ 13.37 (s, 1H, OH), 11.44 (s, 1H, N12H), 11.14 (s, 1H, N2′H), 8.20–8.21 (m, 1H, C8H), 7.86–7.89 (m, 2H, HAr), 7.77–7.83 (m, 5H, C10H + HAr), 7.66–7.68 (m, 2H, HAr), 7.34–7.37 (m, 1H, HAr), 7.07 (d, J = 10.2 Hz, 1H, C1H), 6.98–7.00 (m, 2H), 6.66 (s, 1H, C5′H), 5.72 (d, J = 10.2 Hz, 1H, C2H), 3.74 (s, 3H), 1.38 (s, 3H, C3Me), 1.33 (s, 3H, C3Me).
13C NMR (151 MHz, DMSO-d6) δ 180.97 (C=O), 161.75 (C-OMe), 161.06 (C6), 157.85 (C4a), 157.51 (t, 2JC-F = 22.1 Hz, CF2-CO2H), 152.19 (C-Ar), 149.39 (C-Ar’), 140.92 (C11a), 137.99 (C12a), 134.60 (C10), 134.25 (p-CPh), 129.40 (m-CPh, 2C), 128.05 (o-CPh, 2C), 127.64 (o-CPMP, 2C), 125.73 (C8), 124.90 (C2), 124.81 (i-CPMP + i-CPh, 2C), 122.55 (C9), 118.62 (C7a), 117.88 (C11), 115.50 (C1), 114.22 (m-CPMP, 2C), 105.81 (C5), 102.96 (C6a), 98.01 (C12b), 78.66 (C3), 55.36 (OMe), 41.64 (C5′), 27.94 (C3Me), 27.08 (C3Me), C3F7- group is not registered due to extensive C-F spin–spin coupling.
19F NMR (565 MHz, DMSO-d6) δ −80.11 (t, J = 8.2 Hz), −115.76 (q, J = 8.2 Hz), −126.17.
IR: 1680 (C=O), 1225, 1204, 1146 cm−1.
Calculated, %: C, 59.22; H, 3.79; N, 7.27. Founded, %: C, 59.20; H, 3.76; N, 7.05.
6-Hydroxy-3,3-dimethyl-5-(6-(naphthalen-2-yl)-3-phenyl-2,5-dihydro-1,2,4-triazin-5-yl)-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9d. Yield 51% (403 mg).
1H NMR (400 MHz, DMSO-d6) δ 13.54 (br s, 1H, OH), 11.41 (s, 1H, N12H), 11.35 (br s, 1H, N2′H), 8.48 (d, J = 1.8 Hz, 1H, HAr), 8.19–8.21 (m, 1H, C8H), 8.04 (dd, J = 8.8, J = 1.8 Hz, 1H, HAr), 7.96 (d, J = 8.8 Hz, 1H, HAr), 7.84–7.91 (m, 4H, C10H + HAr), 7.74–7.83 (m, 3H, HAr), 7.68–7.72 (m, 2H, HAr), 7.52–7.58 (m, 2H, HAr), 7.31–7.35 (m, 1H, HAr), 7.05 (d, J = 10.1 Hz, 1H, C1H), 6.88 (s, 1H, C5′H), 5.72 (d, J = 10.1 Hz, 1H, C2H), 1.43 (s, 3H, C3Me), 1.36 (s, 3H, C3Me).
13C NMR (101 MHz, DMSO-d6) δ 180.99 (C=O), 161.24 (C6), 157.79 (C4a), 157.49 (t, 2JC-F = 22.1 Hz, CF2-CO2H), 152.54 (C-Ar), 149.47 (C-Ar), 140.89 (C11a), 138.02 (C12a), 134.53 (C10), 134.33 (p-CPh), 134.02, 132.13, 130.05, 129.41 (m-CPh, 2C), 128.67, 128.39, 128.13 (o-CPh, 2C), 127.93, 127.66, 127.10, 126.22, 125.74 (C8), 124.86 (i-CPh), 124.79 (C1), 122.62, 122.49 (C9), 118.57 (C7a), 117.83 (C11), 115.51 (C1), 105.80 (C5), 102.93 (C6a), 98.08 (C12b), 78.68 (C3), 41.87 (C5′), 27.99 (C3Me), 26.97 (C3Me), C3F7- group is not registered due to extensive C-F spin–spin coupling.
IR: 1683 (C=O), 1226, 1207, 1145 cm−1.
Calculated, %: C, 62.28; H, 3.70; N, 7.09. Founded, %: C, 62.24; H, 3.74; N, 7.12.
19F NMR (565 MHz, DMSO-d6) δ –80.10 (t, J = 8.3 Hz), –115.69 (q, J = 8.3 Hz), –126.15.
5-(3,6-bis(4-Methoxyphenyl)-2,5-dihydro-1,2,4-triazin-5-yl)-6-hydroxy-3,3-dimethyl-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9e. Yield 63% (504 mg).
1H NMR (400 MHz, DMSO-d6) δ 13.23 (br s, 1H, OH), 11.46 (s, 1H, N12H), 10.93 (br s, 1H, N2′H), 8.19–8.21 (m, 1H, C8H), 7.87 (d, J = 8.7 Hz, 2H, PMP), 7.85–7.81 (m, 3H, PMP + C11H), 7.81–7.85 (m, 1H, C10H), 7.32–7.35 (m, 1H, C9H), 7.21 (d, J = 8.7 Hz, 2H, PMP), 7.08 (d, J = 10.1 Hz, 1H, C1H), 6.98 (d, J = 8.6 Hz, 2H, PMP), 6.62 (s, 1H, C5′H), 5.69 (d, 1H, 10.1 Hz, C2H), 3.87 (s, 3H, -OMe), 3.73 (s, 3H, -OMe), 1.34 (s, 6H, C3Me2).
13C NMR (101 MHz, DMSO-d6) δ 180.94 (C=O), 163.89 (C-OMe), 161.63 (C6), 161.06 (C-OMe), 157.83 (C4a), 157.56 (t, 2JC-F = 21.8 Hz, CF2-CO2H), 151.38 (C-PMP), 148.94 (C-PMP’), 140.91 (C11a), 137.95 (C12a), 134.50 (C10), 130.01 (o-CPMP, 2C), 127.56 (o-CPMP, 2C), 125.65 (C8), 124.97 (i-CPMP), 124.77 (C2), 122.47 (C9), 118.61 (C7a), 117.86 (C11), 116.37 (i-CPMP), 115.51 (C1), 114.84 (m-CPMP, 2C), 114.17 (m-CPMP, 2C), 105.87 (C5), 102.96 (C6a), 97.97 (C12b), 78.56 (C3), 55.83 (OMe), 55.32 (OMe), 41.52 (C5′), 27.79 (C3Me), 27.12 (C3Me), C3F7- group is not registered due to extensive spin–spin coupling.
19F NMR (565 MHz, DMSO-d6) δ –80.10 (t, J = 8.1 Hz), –115.68 (q, J = 8.1 Hz), –126.15.
IR: 1680 (C=O), 1257, 1179, 1148 cm−1.
Calculated, %: C, 58.50; H, 3.90; N, 7.00. Founded, %: C, 58.45; H, 3.88; N, 6.91.
6-Hydroxy-3,3-dimethyl-5-(6-phenyl-3-(pyridin-2-yl)-2,5-dihydro-1,2,4-triazin-5-yl)-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9f. Yield 63% (467 mg).
1H NMR (400 MHz, DMSO-d6) δ 13.70 (br s, 1H, OH), 11.48 (br s, 1H, N12H), 11.38 (s, 1H, N2′H), 8.88–8.89 (m, 1H, HAr), 8.18–8.25 (m, 3H, C8H + HAr), 7.92–7.94 (m, 2H, HAr), 7.76–7.85 (m, 3H, C10H + HAr), 7.44–7.45 (m, 3H, HAr), 7.33–7.37 (m, 1H, HAr), 7.04 (d, J = 10.2 Hz, 1H, C1H), 6.73 (s, 1H, C5′H), 5.69 (d, J = 10.2 Hz, 1H, C2H), 1.38 (s, 3H, C3Me), 1.29 (s, 3H, C3Me).
13C NMR (101 MHz, DMSO-d6) δ 180.96 (C=O), 161.38 (C6), 157.88 (C4a), 157.55 (t, 2JC-F = 22.1 Hz, CF2-CO2H), 150.39, 149.83, 149.48, 142.30, 140.88 (C11a), 138.54 (C12a), 138.01 (γ-CPy), 134.51 (C10), 132.73, 131.40, 128.79 (β-CPy), 128.73 (m-CPh, 2C), 125.95 (o-CPh, 2C), 125.60 (C8), 124.77 (C2), 123.50 (β-CPy), 122.48 (C9), 118.61 (C7a), 117.83 (C11), 115.47 (C1), 105.66 (C5), 102.90 (C6a), 97.84 (C12b), 78.65 (C3), 41.80 (C5′), 27.61 (C3Me), 27.25 (C3Me), C3F7- group is not registered due to extensive C-F spin–spin coupling.
19F NMR (376 MHz, DMSO-d6) δ −80.10 (t, J = 8.3 Hz), −115.75 (q, J = 8.3 Hz), –126.14.
IR: 1682 (C=O), 1224, 1203, 1147 cm−1.
Calculated, %: C, 58.30; H, 3.53; N, 9.44. Founded, %: C, 58.16; H, 3.38; N, 9.24.
6-Hydroxy-5-(6-(4-methoxyphenyl)-3-(pyridin-2-yl)-2,5-dihydro-1,2,4-triazin-5-yl)-3,3-dimethyl-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9g. Yield 61% (470 mg).
1H NMR (400 MHz, DMSO-d6) δ 13.62 (br s, 1H, OH), 11.39 (s, 1H, N12H), 11.32 (br s, 1H, N2′H), 8.87–8.88 (m, 1H, HAr), 8.18–8.24 (m, 3H, C8H + HAr), 7.90 (d, J = 8.7, 2H, PMP), 7.77–7.83 (m, 3H, C10H + HAr), 7.34–7.37 (m, 1H, HAr), 7.04 (d, J = 10.0 Hz, 1H, C1H), 6.99 (d, J = 8.7 Hz, 2H, PMP), 6.68 (s, 1H, C5′H), 5.68 (d, J = 10.0 Hz, 1H, C2H), 3.75 (s, 3H), 1.35 (s, 3H, C3Me), 1.28 (s, 3H, C3Me).
13C NMR (101 MHz, DMSO-d6) δ 180.96 (C=O), 161.80 (C6), 161.17 (C-OMe), 157.92 (C4a), 157.57 (t, 2JC-F = 21.8 Hz, CF2-CO2H), 150.36, 149.57, 149.15, 142.37, 140.89 (C11a), 138.51 (C12a), 137.96 (γ-CPy), 134.52 (C10), 128.69 (β-CPy), 127.74 (o-CPMP, 2C), 125.66 (C8), 124.90 (i-CPMP), 124.78 (C2), 123.39 (β-CPy), 122.49 (C9), 118.61 (C7a), 117.84 (C11), 115.47 (C1), 114.20 (m-CPMP, 2C), 105.87 (C5), 102.94 (C6a), 97.90 (C12b), 78.60 (C3), 55.36 (OMe), 41.66 (C5′), 27.60 (C3Me), 27.15 (C3Me), C3F7- group is not registered due to extensive C-F spin–spin coupling.
19F NMR (376 MHz, DMSO-d6) δ –80.10 (t, J = 8.3 Hz), –115.72 (q, J = 8.3 Hz), –126.13.
IR: 1636 (C=O), 1253, 1209, 1176, 1142 cm−1.
Calculated, %: C, 57.59; H, 3.66; N, 9.08. Founded, %: C, 57.63; H, 3.58; N, 9.10.
5-(3-(Ethylthio)-2,5-dihydro-1,2,4-triazin-5-yl)-6-hydroxy-3,3-dimethyl-3,12-dihydro-7H-pyrano[2,3-c]acridin-7-one heptafluorobutanoate 9h. Yield 84% (544 mg).
1H NMR (600 MHz, DMSO-d6) δ 15.94 (s, 1H, CO2H), 12.81 (br s, 1H, OH), 11.50 (s, 1H, NH), 10.71 (br s, 1H, NH), 8.19–8.20 (m, 1H, C8H), 7.85–7.87 (m, 1H, C10H), 7.79–7.82 (m, 1H), 7.34–7.37 (m, 1H), 7.30 (s, 1H), 7.16 (d, J = 10.1 Hz, 1H, C1H), 5.79 (d, J = 10.1 Hz, 1H, C2H), 5.79 (s, 1H, C5′H), 3.25 (dq, J = 14.2, 7.2 Hz, 1H, CHHCH3), 3.17 (dq, J = 14.2, 7.2 Hz, 1H, CHHCH3), 1.50 (s, 3H, C3Me), 1.42 (s, 3H, C3Me), 1.26 (t, J = 7.2 Hz, 3H, CH2CH3).
13C NMR (151 MHz, DMSO-d6) δ 181.00 (C=O), 161.90 (C6), 158.36 (C4a), 158.20 (C3′), 157.63 (t, 2JC-F = 21.8 Hz, CF2-CO2H), 144.30 (C6′), 140.96 (C11a), 138.12 (C12a), 134.56 (C10), 125.98 (C8), 124.88 (C2), 122.51 (C9), 118.69 (C7a), 117.91 (qt, 1JC-F = 287 Hz, 2JC-F = 34.7 Hz, CF3), 117.90 (C11), 115.73 (C1), 109.07 (tq, 1JC-F = 263 Hz, 2JC-F = 35.9 Hz, CF2-CO2H), 104.05 (C5), 103.10 (C6a), 98.22 (C12b), 78.47 (C3), 42.96 (C5′), 27.56 (C3CH3), 26.92 (C3CH3), 24.95 (CH2CH3), 14.52 (CH2CH3), one -CF2- group is not registered due to extensive spin–spin coupling.
19F NMR (565 MHz, DMSO-d6) δ −80.14 (t, J = 8.1 Hz), −115.91 (q, J = 8.1 Hz), −126.21.
IR: 1680 (C=O), 1265, 1208, 1148 cm−1.
Calculated, %: C, 50.00; H, 3.57; N, 8.64. Founded, %: C, 50.12; H, 3.50; N, 8.35.

3.2. Anticancer Activity

  • Cell culture
The studies were carried out on cultured human glioblastoma cells (A172), breast cancer cells (Hs578T), and human embryonic kidney cells (HEK-293) obtained from the shared research facility “Vertebrate cell culture collection” (Institute of Cytology RAS, Russia). All cell lines were grown in Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and maintained in an incubator at 37 °C and 5% CO2-humidified atmosphere. Cells were passaged when reaching ≥ 90% confluence by gentle trypsinization with 0.25% trypsin.
  • Cell viability assay
Stock solutions of the tested compounds of concentration 0.0125 M were prepared by diluting the suspension into an appropriate volume of DMSO, and then diluting the obtained solution in the nutrient medium DMEM/F-12 (10% FBS) to obtain solutions of the tested concentration in the range from 0.5 to 64 μM, so that the effective DMSO concentration did not exceed 1%.
Cells were seeded in 96-well plates at a density of 4 × 103 per well, and incubated overnight at 37 ˚C prior to experimentation. After 24 h, test compounds were added to the wells in a given concentration range. Then the cells were incubated for 72 h, after which a solution of MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (at 5 mg/mL) was added to each well at 20 µL. After 2 h, the medium was removed from the wells. The formazan crystals were solubilized with 200 µL of a mixture of DMSO and isopropanol, 1:1, and the plates were shaken for 10 min. The absorbance was measured at 570 nm using a plate reader.
  • Quantification of apoptotic cell death
The extent of apoptosis and/or necrosis was assayed using an Annexin V-FITC apoptosis detection kit (G1511, Wuhan Servicebio Technology Co., Ltd., Wuhan, China) and propidium iodide (PI). The cells were incubated in 6-well plates and then cultured for 24 h to allow complete attachment to the surface of the plates. Subsequently, solutions of the test compounds and cisplatin were added at an IC50 concentration. After being cultured for 72 h, cells were washed with phosphate-buffered saline (PBS), then cells were stripped with 0.25% trypsin solution, centrifuged at 200 g for 5 min by centrifuge 2–16 KL (Sigma, Hamburg, Germany). The cell precipitate was re-suspended in 25 μL phosphate buffer. To the resulting suspension, 2,5 µL of each dye was added, pipetted, and incubated for 10 min in the dark. After that, 10 μL of the mixture was added to a cell counter slide (C100, RWD, Shenzhen, China) for automatic counting.
  • Mitochondrial membrane potential assay
The determination of transmembrane potential was measured using an anionic dye JC-1 (G1515-100T, Wuhan Servicebio Technology Co., Ltd., Wuhan, China). Cells were pre-dispersed into wells of a 96-well plate at a seeding concentration of 4 × 103 cells per well the day before addition of the test substances. Then, cells with the compounds were cultured for 72 h and incubated with 0.5 mL JC-1 working solution. Cells were incubated at 37 °C and 5% CO2 in a humidified incubator for 30 min. The cells were washed twice with JC-1 buffer solution, and the mitochondrial membrane potential (ΔΨm) of cells was detected with microscope XDS-3FL4 (OPTIKA, Ponteranica, Italy). DPB (1,3-diethylamidazolyl-2-platinate(II) 6-(4-(2-ethylhexyl)oxyphenyl)-2,2′-bipyridine) was used for comparison as a positive control at 0.737 μM concentration for 72 h.
  • Cell proliferative activity
Proliferative activity of cells was assessed using a cell proliferation assay kit Click-iT EdU-555 (G1602, Wuhan Servicebio Technology Co., Ltd., Wuhan, China). EdU (5-ethynyl-2-deoxyuridine) is a nucleoside analog of thymidine and is incorporated into DNA during active synthesis and replication.
Cells were pre-dispersed into the wells of a 96-well plate at a seeding concentration of 4 × 103 cells per well the day before the addition of the tested compounds, and incubated in a CO2 incubator for 24 h. Subsequently, suspensions of the test compounds were added at an IC50 concentration. After 24 h, EdU reagent was added at a 10 μM concentration, and the cultures were incubated for 2 h. Then, the cells were fixed with 10% formaldehyde and permeabilized with 0.5% Triton X-100. The cells were then incubated with the reaction fluorescent mixture iF555, and microscopically examined by XDS-3FL4 (OPTIKA, Ponteranica, Italy).
  • Statistical analysis
Statistical data processing was carried out in the RStudio program (Version 2023.09.1© 2009–2023 RStudio, PBC, Boston, MA, USA) using the R package (version 4.3.2). The cytotoxicity index (IC50) was calculated by plotting dose–response curves using the “drc” package [41].

4. Conclusions

A method for synthesizing a series of pyranoacridone derivatives modified with a 1,2,4-triazine moiety has been developed. The obtained compounds exhibit activity against glioblastoma A172 and breast cancer Hs578T cells in the micromolar range (IC50 value for lead compound 9g is 3.4 μM for glioblastoma A172; selectivity index, 7.59). The addition of the 1,2,4-triazine ring enhances cytotoxicity towards cancer cells and improves selectivity, thus representing a promising approach to improve the antitumor properties of acridones. Antitumor activity of compound 9g was demonstrated against other types of cancer (HCT116 and HuTu 80), and it was found that it is not related to the presence of the heptafluorobutyl anion. Additionally, the mechanism of anticancer activity was studied, revealing that the compound exhibits a non-apoptotic mechanism of anticancer activity.

Supplementary Materials

The following supporting information can be downloaded at https://www.mdpi.com/article/10.3390/ijms27041969/s1.

Author Contributions

Conceptualization, R.F.F. and I.A.K.; software, V.V.M. and I.I.B.; formal analysis, A.D.S., A.P.P. and E.V.N.; investigation, A.A.Z., A.V.P. and R.F.F.; writing—original draft preparation, I.A.K.; writing—review and editing, R.F.F.; funding acquisition, R.F.F. All authors have read and agreed to the published version of the manuscript.

Funding

This work was carried out with the financial support of the Ministry of Science and Higher Education of the Russian Federation (theme FEUZ-2026-0008).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Magalhaes, L.G.; Ferreira, L.L.G.; Andricopulo, A.D. Recent advances and perspectives in cancer drug design. An. Acad. Bras. Ciências 2018, 90, 1233–1250. [Google Scholar] [CrossRef]
  2. Available online: https://www.who.int/news-room/fact-sheets/detail/cancer (accessed on 17 September 2025).
  3. Amin Mir, M.; Banik, B.K. Sustainable healing: Natural compounds facilitating the future cancer treatment. World Dev. Sustain. 2025, 6, 100215. [Google Scholar] [CrossRef]
  4. Shaik, B.B.; Katari, N.K.; Jonnalagadda, S.B. Role of natural products in developing novel anticancer agents: A perspective. Chem. Biodivers. 2022, 19, e202200535. [Google Scholar] [CrossRef] [PubMed]
  5. Altmann, K.-H.; Gertscha, J. Anticancer drugs from nature—Natural products as a unique source of new microtubule-stabilizing agents. Nat. Prod. Rep. 2007, 24, 327–357. [Google Scholar] [CrossRef] [PubMed]
  6. Jones, S. Head-to-head: Docetaxel challenges paclitaxel. Eur. J. Cancer Suppl. 2006, 4, 4–8. [Google Scholar] [CrossRef]
  7. Michael, J.P. Acridone alkaloids. In The Alkaloids: Chemistry and Biology; Elsevier BV: Amsterdam, The Netherlands, 2017; pp. 1–108. [Google Scholar] [CrossRef]
  8. Motiur Rahman, A.F.M.; Liang, J.L.; Lee, S.H.; Son, J.K.; Jung, M.-J.; Kwon, Y.; Jahng, Y. 2,2-Dimethyl-2H-pyran-derived alkaloids I. Practical synthesis of acronycine and benzo[b]acronycine and their biological properties. Arch. Pharm. Res. 2008, 31, 1087–1093. [Google Scholar] [CrossRef]
  9. Kawaii, S.; Tomono, Y.; Katase, E.; Ogawa, K.; Yano, M.; Takemura, Y.; Ju-ichi, M.; Ito, C.; Furukawa, H. The antiproliferative effect of acridone alkaloids on several cancer cell lines. J. Nat. Prod. 1999, 62, 587–589. [Google Scholar] [CrossRef]
  10. Hughes, G.I.; Lahey, F.N.; Price, J.R.; Webb, L.J. Alkaloids of the Australian Rutaceæ. Nature 1948, 162, 223–224. [Google Scholar] [CrossRef]
  11. Yadav, T.T.; Murahari, M.; Peters, G.J.; Mayur, Y.C. A comprehensive review on acridone based derivatives as future anti-cancer agents and their structure activity relationships. Eur. J. Med. Chem. 2022, 239, 114527. [Google Scholar] [CrossRef]
  12. Tillequin, F.; Michel, S.; Skaltsounis, A.-L. Acronycine type alkaloids: Chemistry and biology. In Alkaloids: Chemical and Biological Perspectives; Pelletier, S.W., Ed.; Elsevier: Amsterdam, The Netherlands, 1998; Volume 12, Chapter 1, pp. 1–102. [Google Scholar] [CrossRef]
  13. Scarffe, J.H.; Beaumont, A.R.; Crowther, D. Phase I-II evaluation of acronine in patients with multiple myeloma. Cancer Treat. Rep. 1983, 67, 93–94. [Google Scholar]
  14. Elomri, A.; Mitaku, S.; Michel, S.; Skaltsounis, A.-L.; Tillequin, F.; Koch, M.; Pierré, A.; Guilbaud, N.; Léonce, S.; Kraus-Berthier, L.; et al. Synthesis and cytotoxic and antitumor activity of esters in the 1,2-dihydroxy-1,2-dihydroacronycine series. J. Med. Chem. 1996, 39, 4762–4766. [Google Scholar] [CrossRef] [PubMed]
  15. Magiatis, P.; Mitaku, S.; Skaltsounis, A.-L.; Tillequin, F.; Koch, M.; Pierré, A.; Atassi, G. Synthesis and biological activity of esters in the trans-1,2-dihydroxy-1,2-dihydroacronycine series. J. Nat. Prod. 1998, 61, 198–201. [Google Scholar] [CrossRef] [PubMed]
  16. Costes, N.; Michel, S.; Tillequin, F.; Koch, M.; Pierré, A.; Atassi, G. Chiral dihydroxylation of acronycine: Absolute configuration of natural cis-1,2-dihydroxy-1,2-dihydroacronycine and cytotoxicity of (1R,2R)-and (1S,2S)-1,2-diacetoxy-1,2-dihydroacronycine. J. Nat. Prod. 1999, 62, 490–492. [Google Scholar] [CrossRef] [PubMed]
  17. Do, Q.; Thi Mai, H.D.; Gaslonde, T.; Pfeiffer, B.; Léonce, S.; Pierré, A.; Michel, S.; Tillequin, F.; Dufat, H. Structure-activity relationships in the acronycine and benzo[b]acronycine series: Role of the pyran ring. Eur. J. Med. Chem. 2008, 43, 2677–2687. [Google Scholar] [CrossRef]
  18. Nguyen, Q.C.; Nguyen, T.T.; Yougnia, R.; Gaslonde, T.; Dufat, H.; Michel, S.; Tillequin, F. Acronycine derivatives: A promising series of anti-cancer agents. Anti-Cancer Agents Med. Chem. 2009, 9, 804–815. [Google Scholar] [CrossRef]
  19. Banerjee, J.; Kundu, I.; Zhang, S.; Wen, S.; Chattopadhyay, S.K. Synthesis and preliminary biophysical and cellular evaluation of some ring-enlarged analogues of the anti-tumor plant alkaloid acronycine. ACS Omega 2019, 4, 6106–6113. [Google Scholar] [CrossRef]
  20. Doan Thi Mai, H.; Gaslonde, T.; Michel, S.; Tillequin, F.; Koch, M.; Bongui, J.-B.; Elomri, A.; Seguin, E.; Pfeiffer, B.; Renard, P.; et al. Structure activity relationships and mechanism of action of antitumor benzo[b]pyrano [3,2-h]acridin-7-one acronycine analogs. J. Med. Chem. 2003, 46, 3072–3082. [Google Scholar] [CrossRef]
  21. David-Cordonnier, M.-H.; Laine, W.; Lansiaux, A.; Kouach, M.; Briand, G.; Pierré, A.; Hickman, J.A.; Bailly, C. Alkylation of guanine by S-23906-1, a novel potent antitumor compound derived from the plant alkaloid acronycine. Biochemistry 2002, 41, 9911–9920. [Google Scholar] [CrossRef]
  22. Santra, S.; Sharapov, A.D.; Fatykhov, R.F.; Potapova, A.P.; Khalymbadzha, I.A.; Valieva, M.I.; Kopchuk, D.S.; Zyryanov, G.V.; Bunev, A.S.; Melekhin, V.V.; et al. Xanthone-1,2,4-triazine and acridone-1,2,4-triazine conjugates: Synthesis and anticancer activity. Pharmaceuticals 2023, 16, 403. [Google Scholar] [CrossRef]
  23. Hari, G.S.; Lee, Y.R.; Wang, X.; Lyoo, W.S.; Kim, S.H. New Synthetic Routes to Acronycine, Noracronycine, and Their Analogues. Bull. Korean Chem. Soc. 2010, 31, 2406–2409. [Google Scholar] [CrossRef]
  24. Fatykhov, R.F.; Inyutina, A.K.; Khalymbadzha, I.A.; Sharapov, A.D.; Bobkina, M.V.; Chupakhin, O.N. Optimization of pyran ring annulation in the synthesis of acronycine and its derivatives. AIP Conf. Proc. 2019, 2063, 040014. [Google Scholar] [CrossRef]
  25. Khalymbadzha, I.A.; Fatykhov, R.F.; Butorin, I.I.; Sharapov, A.D.; Potapova, A.P.; Muthipeedika, N.J.; Zyryanov, G.V.; Melekhin, V.V.; Tokhtueva, M.D.; Deev, S.L.; et al. Bioinspired pyrano [2,3-f]chromen-8-ones: Ring C-opened analogues of Calanolide A: Synthesis and anti-HIV-1 evaluation. Biomimetics 2024, 9, 44. [Google Scholar] [CrossRef] [PubMed]
  26. Khalymbadzha, I.A.; Chupakhin, O.N.; Fatykhov, R.F.; Charushin, V.N.; Schepochkin, A.V.; Kartsev, V.G. Transition-metal-free cross-dehydrogenative coupling of triazines with 5,7-dihydroxycoumarins. Synlett 2016, 27, 2606–2610. [Google Scholar] [CrossRef]
  27. Potapova, A.P.; Taniya, O.S.; Valieva, M.I.; Khalymbadzha, I.A.; Sharapov, A.D.; Starnovskaya, E.S.; Kopchuk, D.S.; Gaviko, V.S.; Novikov, A.S.; Kalinichev, A.A.; et al. Synthesis and photophysical study of novel 3-triazinyl- and 3-pyridinyl-substituted coumarins. Opt. Mater. 2025, 158, 116471. [Google Scholar] [CrossRef]
  28. Fatykhov, R.F.; Khalymbadzha, I.A.; Sharapov, A.D.; Potapova, A.P.; Tsmokalyuk, A.N.; Gaviko, V.S. Heptafluorobutyric acid catalyzed cross-dehydrogenative coupling of 7-aminocoumarins with 1,2,4-triazines: A straightforward pathway to 3-triazinyl-7-aminocoumarins. Synthesis 2024, 56, 1958–1966. [Google Scholar] [CrossRef]
  29. Fatykhov, R.F.; Savchuk, M.I.; Starnovskaya, E.S.; Bobkina, M.V.; Kopchuk, D.S.; Nosova, E.V.; Zyryanov, G.V.; Khalymbadzha, I.A.; Chupakhin, O.N.; Charushin, V.N.; et al. Nucleophilic substitution of hydrogen–the Boger reaction sequence as an approach towards 8-(pyridin-2-yl)coumarins. Mendeleev Commun. 2019, 29, 299–300. [Google Scholar] [CrossRef]
  30. Fatykhov, R.F.; Khalymbadzha, I.A.; Sharapov, A.D.; Potapova, A.P.; Mochulskaya, N.N.; Tsmokalyuk, A.N.; Ivoilova, A.V.; Mozharovskaia, P.N.; Santra, S.; Chupakhin, O.N. MnO2-mediated oxidative cyclization of “formal” schiff’s bases: Easy access to diverse naphthofuro-annulated triazines. Molecules 2022, 27, 7105. [Google Scholar] [CrossRef]
  31. Kleszczyński, K.; Składanowski, A.C. Mechanism of cytotoxic action of perfluorinated acids. III. Disturbance in Ca2+ homeostasis. Toxicol. Appl. Pharmacol. 2011, 251, 163–168. [Google Scholar] [CrossRef]
  32. Kaczmarska, I.; Mokra, K.; Michałowicz, J. Perfluorooctanoic acid and its short-chain substitutes induce cytotoxic and prooxidative changes in human peripheral blood mononuclear cells: A comparative study. Int. J. Mol. Sci. 2025, 26, 5408. [Google Scholar] [CrossRef]
  33. Henry, N.D.; Fair, P.A. Comparison of in vitro cytotoxicity, estrogenicity and anti-estrogenicity of triclosan, perfluorooctane sulfonate and perfluorooctanoic acid. J. Appl. Toxicol. 2013, 33, 265–272. [Google Scholar] [CrossRef]
  34. Kozurkova, M. Acridine derivatives as inhibitors/poisons of topoisomerase II. J. Appl. Toxicol. 2022, 42, 544–552. [Google Scholar] [CrossRef]
  35. Iwona, G. ‘Acridines’ as new horizons in antifungal treatment. Molecules 2020, 25, 1480. [Google Scholar] [CrossRef]
  36. Herole, R.; Rakesh, K.; Rajendra, D. A review on acridine, xanthene and its derivatives: Synthesis, physical and pharmacological properties. Trop. J. Pharm. Life Sci. 2022, 9, 1–12. [Google Scholar] [CrossRef]
  37. Liang, X.; Wu, Q.; Luan, S.; Yin, Z.; He, C.; Yin, L.; Zou, Y.; Yuan, Z.; Li, L.; Song, X.; et al. A comprehensive review of topoisomerase inhibitors as anticancer agents in the past decade. Eur. J. Med. Chem. 2019, 171, 129–168. [Google Scholar] [CrossRef]
  38. Urvashee, G.; Kalyani, P.; Riya, S.; Manash, P.; Tirna, P.; Shah, A.; Aparoop, D. Recent advances on natural and non-natural xanthones as potential anticancer agents: A review. Med. Chem. 2023, 19, 757–784. [Google Scholar] [CrossRef] [PubMed]
  39. Gurjar, V.K. Recent advances in oxygen-containing heterocycles as anticancer agents. Int. J. Med. Pharm. Health Sci. 2024, 2, 80–93. [Google Scholar] [CrossRef]
  40. Reisch, J.; Herath, H.M.T.B.; Kumar, N.S. Natural product chemistry, 143. Convenient synthesis of isoacronycine and some other new acridone derivatives. Justus Liebigs Ann. Chem. 1991, 1991, 685–689. [Google Scholar] [CrossRef]
  41. Ritz, C.; Baty, F.; Streibig, J.C.; Gerhard, D. Dose-response analysis using R. PLoS ONE 2015, 10, e0146021. [Google Scholar] [CrossRef]
Ijms 27 01969 g001
Figure 1. Acronycine and its modified analogs with cytotoxic activity (IC50) against L1210 cell lines, μM.
Figure 1. Acronycine and its modified analogs with cytotoxic activity (IC50) against L1210 cell lines, μM.
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Figure 2. Comparison of our previous (blue) and present work (magenta).
Figure 2. Comparison of our previous (blue) and present work (magenta).
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Scheme 1. Possible pathways to compound 9. Blue arrows – initial strategy A for synthesis of 9; Red arrows – alternative strategy B.
Scheme 1. Possible pathways to compound 9. Blue arrows – initial strategy A for synthesis of 9; Red arrows – alternative strategy B.
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Scheme 2. Synthesized noracronycine derivatives.
Scheme 2. Synthesized noracronycine derivatives.
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Figure 3. Concentration–viability response curves of compounds 9b,d,e,g and 10 on A172, HEK-293, and Hs578T, mean ± SD.
Figure 3. Concentration–viability response curves of compounds 9b,d,e,g and 10 on A172, HEK-293, and Hs578T, mean ± SD.
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Figure 4. Comparison of the cytotoxicity index of the studied compounds in malignant cell cultures A172, Hs578T, and normal HEK293 cells, IC50 ± SE.
Figure 4. Comparison of the cytotoxicity index of the studied compounds in malignant cell cultures A172, Hs578T, and normal HEK293 cells, IC50 ± SE.
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Figure 5. Number of apoptotic cells for the A172 line after 72 h incubation with cisplatin and 9g at IC50 concentration.
Figure 5. Number of apoptotic cells for the A172 line after 72 h incubation with cisplatin and 9g at IC50 concentration.
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Figure 6. Staining of A172 cells with JC-1 dye. “Control +”—cells treated with DPB (1,3-diethylamidazolyl-2-platinate (II) 6-(4-(2-ethylhexyl)oxyphenyl)-2,2′-bipyridine) at 0.737 μM concentration for 72 h before staining (positive control), “9g”, and “Cisplatin”—cells after 24 h incubation with the test compound and cisplatin at an IC50 concentration, respectively. JC-1 produces green (in case of a decrease in ΔΨm of mitochondria) or red (if ΔΨm is normal) fluorescence. On the left are images obtained from a microscope, magnification 200×, on the right is a bar graph reflecting the ratio of the intensity of red and green fluorescence.
Figure 6. Staining of A172 cells with JC-1 dye. “Control +”—cells treated with DPB (1,3-diethylamidazolyl-2-platinate (II) 6-(4-(2-ethylhexyl)oxyphenyl)-2,2′-bipyridine) at 0.737 μM concentration for 72 h before staining (positive control), “9g”, and “Cisplatin”—cells after 24 h incubation with the test compound and cisplatin at an IC50 concentration, respectively. JC-1 produces green (in case of a decrease in ΔΨm of mitochondria) or red (if ΔΨm is normal) fluorescence. On the left are images obtained from a microscope, magnification 200×, on the right is a bar graph reflecting the ratio of the intensity of red and green fluorescence.
Ijms 27 01969 g006
Ijms 27 01969 g007
Figure 7. Staining of A172 cells with the fluorescent probe iF555. “Control −”—intact cells, “9g”, and “Control +”—cells after 24 h incubation with the studied compound and cisplatin at an IC50 concentration, respectively. (A)—bar graph showing the total number of cells, (B)—bar graph showing the percentage of cells in the stage of active DNA replication, (C)—images obtained with a microscope, magnification 100×.
Figure 7. Staining of A172 cells with the fluorescent probe iF555. “Control −”—intact cells, “9g”, and “Control +”—cells after 24 h incubation with the studied compound and cisplatin at an IC50 concentration, respectively. (A)—bar graph showing the total number of cells, (B)—bar graph showing the percentage of cells in the stage of active DNA replication, (C)—images obtained with a microscope, magnification 100×.
Ijms 27 01969 g007
Table 1. Optimization of the reaction conditions.
Table 1. Optimization of the reaction conditions.
Ijms 27 01969 i001
EntryConditionsYield, %
1MsOH (3 equiv.), AcOH, 80 °C, 2 h24
2BF3·Et2O (8 equiv.), MeOH, reflux, 2 h-
3TFA, reflux, 1 h-
4HFBA (3 equiv.), AcOH, 60 °C, 30 min.60
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MDPI and ACS Style

Zonov, A.A.; Fatykhov, R.F.; Khalymbadzha, I.A.; Sharapov, A.D.; Potapova, A.P.; Butorin, I.I.; Melekhin, V.V.; Paramonova, A.V.; Nosova, E.V. Anticancer Acridones, Part 2—Acronycine-Type Derivatives Modified with 2,5-Dihydro-1,2,4-Triazine Moiety: Synthesis and In Vitro Evaluation. Int. J. Mol. Sci. 2026, 27, 1969. https://doi.org/10.3390/ijms27041969

AMA Style

Zonov AA, Fatykhov RF, Khalymbadzha IA, Sharapov AD, Potapova AP, Butorin II, Melekhin VV, Paramonova AV, Nosova EV. Anticancer Acridones, Part 2—Acronycine-Type Derivatives Modified with 2,5-Dihydro-1,2,4-Triazine Moiety: Synthesis and In Vitro Evaluation. International Journal of Molecular Sciences. 2026; 27(4):1969. https://doi.org/10.3390/ijms27041969

Chicago/Turabian Style

Zonov, Andrey A., Ramil F. Fatykhov, Igor A. Khalymbadzha, Ainur D. Sharapov, Anastasia P. Potapova, Ilya I. Butorin, Vsevolod V. Melekhin, Anastasia V. Paramonova, and Emiliya V. Nosova. 2026. "Anticancer Acridones, Part 2—Acronycine-Type Derivatives Modified with 2,5-Dihydro-1,2,4-Triazine Moiety: Synthesis and In Vitro Evaluation" International Journal of Molecular Sciences 27, no. 4: 1969. https://doi.org/10.3390/ijms27041969

APA Style

Zonov, A. A., Fatykhov, R. F., Khalymbadzha, I. A., Sharapov, A. D., Potapova, A. P., Butorin, I. I., Melekhin, V. V., Paramonova, A. V., & Nosova, E. V. (2026). Anticancer Acridones, Part 2—Acronycine-Type Derivatives Modified with 2,5-Dihydro-1,2,4-Triazine Moiety: Synthesis and In Vitro Evaluation. International Journal of Molecular Sciences, 27(4), 1969. https://doi.org/10.3390/ijms27041969

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