Indenyl-thiazole and indenyl-formazan derivatives: Synthesis, anticancer screening studies, molecular-docking, and pharmacokinetic/ molin-spiration properties

Abstract

Two new series of thiazole and formazan linked to 5-Bromo-indan were synthesized, and their structures were assured based on all possible analytical techniques. The size of the tested derivatives was calculated from the XRD technique and found five derivatives 3, 10a, 14a, 15, and 16 on the nanosized scale. The two series were tested for their efficacy and toxicity as anti-colon and stomach cancers. Derivative 10d showed activity more than the two reference drugs used in the case of SNU-16. Surpislly, in the case of COLO205, five derivatives 4, 6c, 6d, 6e, and 10a are better than the two benchmarks used, and two derivatives, 14a and 14b more potent than cisplatin. All potent derivatives showed a strong fit with the active site of the two tested proteins (gastric cancer (PDB = 2BID) and colon cancer (PDB = 2A4L)) in the molecular docking study. The Pharmacophore and ADME studies of the new derivatives showed that most derivatives revealed promising bioactivity, which indicates the drug-likeness properties against kinase inhibitors, protease, and enzyme inhibitors. In addition, the ProTox-II showed that the four compounds 10d, 16, 6d, and 10a are predicted to have oral LD₅₀ values ranging from 335 to 3500 mg/kg in a rat model with (1 s,4 s)-Eucalyptol bearing the highest values and quercetin holding the lowest one.

1. Introduction

Different types of cancer are considered the most dangerous and deadly diseases globally [1, 2]. Numerous attempts have been made to find suitable ways to treat cancer. Chemotherapy is the primary approach in cancer treatment, where various natural and synthetic compounds are used to destroy cancer cells [3]. Despite the rapid advances in pharmacology and chemotherapeutic agents, the treatment of cancers remains a serious problem due to the toxicity, resistance, and lack selectivity of currently available anticancer drugs [4]. So far, the search is still going on for new compounds that are suitable and effective in treating different types of cancer [5–8]. On the other hand, nanoscale synthesis of heterocyclic materials is a scientific achievement due to its amazing applications in all medical and industrial fields [9, 10]. Heterocyclic-nanocomposites have effective biological activities due to their small size, allowing easy penetration of viral or microbial cell membranes [11, 12]. By browsing the heterocyclic compounds, we noted that thiazole derivatives spread in natural and industrial medicines, as they are a major part of cancer, histamine, microbes, and pressure medicines [13–15]. Several thiazoles with antiviral [16], antioxidant [17], antimicrobial [18], anticonvulsant [19], anti-inflammatory [20], and neuroprotective activities [21] are reported.

Fig 1 collected some drugs containing thiazole moiety. Another treasure of organic compounds is formazan derivatives with the general formula (-N = NC = NN -) [22] have been widely studied due to their numerous applications in the health and industrial field [23]. Multiple studies of the synthesis and applications of formazan derivatives showed that they have potent effects as antiparkinsonian [24], antiviral [25], antibacterial [26], antioxidant [27], anti-inflammatory [28], and anticancer activities [29]. Cyclic formazan derivatives are good indicators, especially for lithium-ion in blood [30]. Combining all the above findings with our research project concerning the synthesis of bioactive heterocyclic compounds [31–35], we focused on synthesizing a new series of thiazole and formazan derivatives and then investigating their anti-gastric and anti-colon cancer activities.

[Figure omitted. See PDF.]

Furthermore, several previous studies of compounds containing thiazole moiety proved the effectiveness of these compounds as anticancer drugs [36–40]. The scenario of cancer is one of the most important in medicinal chemistry, while thiazoles are very important in treating cancers. It was reported the importance of the thiazole ring as a scaffold present in a wide range of therapeutic agents, the medicinal chemists have been encouraged to synthesize a large number of novel antitumors bearing this heterocycle, which furnish extensive synthetic possibilities due to the presence of several reaction sites [41–45].

By the way, the insilico-studies gave a chance to check the activities of these compounds, molecular docking studies with MOE and pharmacokinetics studies by molinspiration are good ways to discover and confirm the biological activities of the compounds under investigation [46–48].

2. Experimental

2.1. Chemistry

Instruments (See S1 File)

2.2. Synthesis of 2-(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazine-1-carbothioamide (3)

In a 50 mL round Q.F. flask, we added 0.01 mole of 5-Bromo-indan-1-one 1 (2.11 g) with the same number of moles of thiosemicarbazide 2 (≈1g) in absolute ethanol (30 mL). The mixture was heated under reflux to dissolve the thiosemicarbazide; then, 1mL of concentrated HCl was added, and the reflux was completed to 2 h. The solid thiosemicarbazone 3 was filtered after the solution cold and crystallized from ethanol/dioxane to afford compound 3 as pale yellow, yield: 93%; M.p: 227–230°C; IR: 3430, 3366, 3147 (NH₂, NH), 2917 (sp³-CH), 1593 (C = N), 1464, 1289, 1171 cm^-1, ¹H NMR (850 MHz, CDCl₃): 2.84 (d, J = 8.5 Hz, 2H, CH₂), 3.15–3.20 (m, 2H, CH₂), 6.44 (s, 1H, NH), 7.33 (s, 1H, NH), 7.44 (d, J = 8.5 Hz, 1H, ArH), 7.54(s, 1H, ArH), 7.74 (d, J = 8.5 Hz, 1H, ArH), 8.69 (s, 1H, NH). ¹³C NMR (CDCl₃): 26.7 (CH₂), 28.3 (CH₂), 122.9, 125.7, 128.9, 130.8, 135.8, 150.4, 156.5, 178.7 (C = S). MS(m/z): 285 (M⁺ +2, 29), 284 (M⁺ +1, 6), 283 (M⁺, 27), 268 (15), 266 (14), 129 (10), 115 (67), 102 (31), 90 (8), 89 (42), 76 (39), 60 (100). Elemental Analysis (%): (C₁₀H₁₀BrN₃S; Mwt: 284.18) Calc. (Found): C, 42.27 (42.18); H, 3.55 (3.45); N, 14.79 (14.59)%.

2.3. Synthesis of (5-Bromo-indan-1-ylidene)-hydrazine (4)

In a 25 mL round Q.F. flask, we added 0.005 moles of 5-Bromo-indan-1-one 1 (1.06 g) in 20 mL EtOH and 1mL of NH₂NH₂.H₂O were added, followed by refluxing the whale mixture for 5h. After the reaction (TLC mentoring), the pale-yellow solid was collected and washed with methanol to afford pale yellow crystals, yielding 78%; M.p: 120°C. IR: 3344, 3250 (NH₂), 2918 (sp³-CH), 1469, 1351, 1286, 1169, 1107, 1055 cm^-1. ¹H NMR (850 MHz, CDCl₃) ¹H NMR(CDCl₃): 2.8–2.7 (m, 2H, CH₂), 3.0–3.11 (m, 2H, CH₂), 5.20 (s, 2H, NH₂), 7.37(d, J = 8.5 Hz, 1H, ArH), 7.45 (s, 1H, ArH), 7.5 (d, J = 8.5 Hz, 1H, ArH). ¹³C NMR (CDCl₃): 24.9 (CH₂), 28.1(CH₂), 122.0, 123.1, 128.5, 130.3, 137.7, 148.6, 155.5. MS(m/z): 225 (M⁺, 2), 208 (13), 143 (7), 129 (37), 115 (100), 113 (10), 102 (40), 89 (20), 83 (14), 77 (15), 75 (17), 71 (20), 63 (24), 57 (32), 55 (27). Elemental Analysis (%): (C₉H₉BrN₂; Mwt: 225.09) Calc. (Found): C, 48.02 (47.91); H, 4.03 (4.10); N, 12.45 (12.32)%.

The reaction of 2-(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazine-1-carbothioamide (3) or (5-Bromo-indan-1-ylidene)-hydrazine (4) with hydrazonoyl chlorides 5 or 13 or 13 and phenacyl bromide derivatives 9

The method for synthesis of derivatives 6a-e, 10a-d, 14a, b, 15, and 16 as illustrated in Fig 2.

[Figure omitted. See PDF.]

2-(2h4(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazineyl)-4-methyl-5-(phenyldiazenyl)-thiazole (6a)

Orange solid, Yield: 88%; M.p: 220–222°C; IR: 3113 (NH), 3036 (sp²-CH), 2918 (sp³-CH), 1615 (C = N), 1546, 1410, 1378, 1252, 1169, 1117 cm^-1. ¹H NMR (850 MHz, CDCl₃) 2.69 (s, 3H, CH₃), 3.12–3.21 (m, 4H, 2CH₂), 7.06 (t, J = 7.65Hz, 1H, ArH), 7.23 (d, J = 8.5 Hz, 2H, ArH), 7.37 (t, J = 6.8 Hz, 2H, ArH), 7.44 (s, 1H, ArH), 7.48 (d, J = 7.65 Hz, 1H, ArH), 7.57 (s, 1H, NH), 7.82 (d, J = 7.65 Hz, 1H, ArH). MS(m/z): 427 (M⁺ +1, 29), 426 (M⁺, 11), 425 (29), 285 (4), 129 (27), 115 (71), 112 (10), 102 (24), 92 (19), 77 (100), 65 (29). Elemental Analysis (%): (C₁₉H₁₆BrN₅S; Mwt: 426.34) Calc. (Found): C, 53.53 (53.29); H, 3.78 (3.69); N, 16.43 (16.37)%.

2-(2-(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazineyl)-4-methyl-5-(m-tolyldiazenyl)-thiazole (6b)

Green solid, Yield: 85%; M.p: 128–129°C; IR: 3400 (NH), 3048 (sp²-CH), 2918 (sp³-CH), 1595 (C = N), 1506, 1266, 1189, 1057 cm^-1. ¹H NMR(850 MHz, CDCl₃): 2.36 (s, 3H, CH₃), 2.39 (s, 3H, CH₃), 2.66–3.16 (m, 4H, 2CH₂), 6.87–7.57 (m, 7H, ArH), 7.62 (s, 1H, NH). ¹³C NMR (CDCl₃): 18.4 (CH₃), 21.5 (CH₃), 28.4 (CH₂), 29.6 (CH₂), 111.4, 114.7, 124.1, 124.9, 129.0, 129.3, 130.0, 130.5, 130.9, 134.1, 139.5, 143.2, 156.7, 170.0, 174.1. 177.5. MS(m/z): 440 (M⁺, 3), 439 (5), 129 (19), 115 (53), 106 (18), 102 (20), 91 (100), 78 (10), 77 (27), 65 (24), 51 (7). Elemental Analysis (%): (C₂₀H₁₈BrN₅S; Mwt 440.36) Calc. (Found): C, 54.55 (54.36); H, 4.12 (4.04); N, 15.90 (15.82)%.

2-(2-(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazineyl)-4-methyl-5-(p-tolyldiazenyl)thiazole (6c)

Orange solid, Yield: 87%; M.p: 130–132°C; IR: 3213 (NH), 3181 (sp²-CH), 2917 (sp³-CH), 1612 (C = N), 1500, 1281, 1171, 1117 cm^-1. ¹H NMR(850 MHz, CDCl₃): 2.35 (s, 3H, CH₃), 2.39 (s,3H, CH₃), 2.67–3.19 (m, 4H, 2CH2), 7.12–7.48 (m, 6H, ArH), 7.56 (s,1H, NH), 7.81 (d, J = 8.5Hz, 1H, ArH). ¹³C NMR (CDCl₃): 16.7 (CH₃), 22.7 (CH₃), 28.5 (CH₂), 29.7 (CH₂), 114.1, 124.1, 126.5, 128.1, 129.0, 130.0, 132.6, 136.7, 140.2, 152.7, 174.1, 177.4, (two carbons overlapped). MS(m/z): 440 (M⁺, 3), 135 (5), 129 (22), 115 (58), 106 (18), 102 (20), 91 (100), 77(26), 65 (20), 51 (8). Elemental Analysis (%): (C₂₀H₁₈BrN₅S; Mwt: 440.36) Calc. (Found): C, 54.55 (54.42); H, 4.12 (4.10); N, 15.90 (15.79)%.

2-(2-(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazineyl)-5-((3-chlorophenyl)-diazenyl)-4-methylthiazole (6d)

Red solid, Yield: 89%; M.p: 180°C; IR: 3400 (br. NH), 1600 (C = N), 1508, 1468, 1374, 1242,1173, 1081. ¹H NMR (DMSO-d₆) 2.56 (s, 3H, CH₃), 3.02–3.09 (m, 4H, 2CH₂), 6.99–7.62 (m, 7H, ArH), 7.64 (s, 1H, NH). ¹³C NMR (DMSO-d₆): 16.9 (CH₃), 28.5 (CH₂), 29.5 (CH₂), 114.1, 123.9, 122.0, 123.9, 125.0, 126.1, 129.6, 130.5, 130.9, 131.0, 131.3, 134.3, 136.4, 136.8, 153.6, 158.0. MS(m/z): 461 (M⁺+2, 4), 460 (M⁺+1, 2), 459 (M+, 3), 183 (5), 139 (6), 129 (34), 115 (100), 111 (92), 102 (37), 99 (29), 95 (5), 89 (20), 77 (14), 75 (36), 67 (37), 63 (27), 57 (7), 55 (7), 51 (11). Elemental Analysis (%): (C₁₉H₁₅BrClN₅S; Mwt: 460.78) Calc. (Found): C, 49.53 (49.48); H, 3.28 (3.19); N, 15.20 (15.08)%.

2-(2-(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazineyl)-5-((4-chlorophenyl)diazenyl)-4-methylthiazole (6e)

Red solid, Yield: 83%; M.p: 177–178°C; IR: 3305 (NH), 3174 (sp²-CH), 2919 (sp³-CH), 1598 (C = N), 1487, 1299, 1169, 1024 cm^-1. ¹H NMR(850 MHz, CDCl₃): 1.26–1.36 (m, 2H, CH₂), 2.68(s, 3H, CH₃), 3.13–3.20 (m, 2H, CH₂), 7.16 (d, J = 8.5 Hz, 2H, ArH), 7.32 (d, J = 8.5Hz, 1H, ArH), 7.40 (s, 1H, ArH), 7.47 (d, J = 8.5 Hz, 2H, ArH), 7.57 (s, 1H, NH), 7.80 (d, J = 8.5Hz, 1H, ArH). ¹³C NMR (CDCl₃): 16.7 (CH₃), 27.7 (CH₂), 28.5(CH₂), 115.2, 124.1, 126.7, 127.7, 129.0, 129.5, 130.6, 136.6, 141.2, 141.6, 152.8, 169.5, 174.6, 177.5. MS(m/z): 460 (M⁺, 1), 459 (2), 210 (3), 141 (5), 129 (34), 115 (96), 111 (100), 102 (38), 99 (29), 89 (19), 77 (11), 75 (33), 71 (14), 67 (32), 63(22), 51(9). Elemental Analysis (%): (C₁₉H₁₅BrClN₅S; Mwt: 460.78). Calc C, 49.53; H, 3.28; Br, 17.34; Cl, 7.69; N, 15.20; S, 6.96.

N-(5-Bromo-indan-1-ylidene)-N’-(4-phenyl-thiazol-2-yl)-hydrazine (10a)

Gray solid, Yield: 89%; M.p: 247–248°C; IR: 3400 (NH), 3052 (sp²-CH), 1616 (C = N), 1512, 1435, 1370, 1208, 1117, 1016 cm^-1. ¹H NMR (850 MHz, DMSO-d₆): 2.88 (Br.s, 2H,CH₂), 3.09 (Br.s, 2H,CH₂), 7.32 (s, 1H, thiazole-H), 7.39–7.53 (m, 7H, ArH), 7.60 (s, 1H, NH), 7.88 (d, J = 8.5Hz, 1H, ArH). ¹³C NMR (DMSO-d₆): 28.2 (CH₂), 28.5 (CH₂), 108.1, 122.8, 123.8, 126.0, 128.0, 128.6, 128.8, 129.2, 130.6, 133.4, 137.3, 150.9, 156.3, 165.8. MS(m/z): 384 (M⁺, 12), 383 (24), 210 (4), 176 (45), 148 (9), 134 (100), 121 (12), 115 (24), 102 (30), 89 (17), 77 (20), 63 (8), 51 (9). Elemental Analysis (%): (C₁₈H₁₄BrN₃S; Mwt: 384.30). Calc. (Found) C, 56.26 (56.09); H, 3.67 (3.56); N, 10.93 (10.84)%.

N-(5-Bromo-indan-1-ylidene)-N’-(4-p-tolyl-thiazol-2-yl)-hydrazine (10b)

Gray solid, Yield: 90%; M.p: 233–234°C; IR: 3200 (NH), 3065 (sp²-CH), 2917 (sp³-CH), 1613 (C = N), 1508, 1468, 1205, 1172, 1060, 1016 cm^-1. ¹H NMR (850 MHz, DMSO-d₆): 2.32 (s, 3H, CH₃), 2.89–3.11 (m, 4H, 2CH₂), 7. 22–7.77 (m, 9H, ArH, thiazole-H and NH). ¹³C NMR (DMSO-d₆): 21.2 (CH₃), 28.0 (CH₂), 28.5 (CH₂), 103.4, 122.7, 123.5, 125.9, 128.0, 1 29.1, 129.4 129.6, 130.6, 130.7, 137.3, 137.6, 150.7, 169.6. Mass (m/z): 400 (M⁺ +2, 22), 398 (M⁺, 44), 190(51), 162(10), 148(100), 129(29), 115(52), 102(39), 89(20), 75(10). MS(m/z): 399(M⁺ +2, 88), 397 (M⁺+1, 87), 339 (M+, 4), 289(5), 190(51), 174(9), 162(10), 148(100), 135(10), 129(29), 115(53), 102(38), 91(25), 77(18), 63(15), 51(11). Elemental Analysis (%): (C₁₉H₁₆BrN₃S; Mwt: 398.32). Calc. (Found): C, 57.29 (57.16); H, 4.05 (3.93); N, 10.55 (10.43)%.

N-(5-Bromo-indan-1-ylidene)-N’-[4-(4-bromo-phenyl)-thiazol-2-yl]-hydrazine (10c)

Gray solid, Yield: 91%; M.p: 230–232°C; IR: 3200 (NH), 3048 (sp²-CH), 2919 (sp³-CH), 1610 (C = N), 1491, 1368, 1205, 1104, 1001, 824 cm^-1. ¹H NMR (850 MHz, DMSO-d₆): 2.88 (t, J = 8.5Hz, 2H₁ CH₂), 3.10 (t, J = 8.5 Hz, 2H, CH₂), 7.41 (s, 1H, thiazole-H) 7.5–7.83 (m, 8H, Ar-H and NH).¹³C NMR (DMSO-d₆): 28.0 (CH₂), 28.4 (CH₂), 105.2, 120.9, 122.7, 123.5, 128.0, 129.1, 1 30.6, 132.0 134.4, 137.6, 150.7, 155.3, 169.9. MS(m/z): 463 (M⁺, 25), 461 (15), 256 (50), 210 (17), 182 (5), 174 (100), 155 (5), 146 (25), 133 (17), 129 (74), 120 (25), 115 (79), 102 (87), 95 (5), 89 (44), 75 (32), 63 (22), 57 (10), 51 (20). Elemental Analysis (%): (C₁₈H₁₃Br₂N₃S; Mwt: 463.19). Calc. (Found): C, 46.68 (46.52); H, 2.83 (2.78); N, 9.07 (9.01)%.

N-(5-Bromo-indan-1-ylidene)-N’-[4-(4-chloro-phenyl)-thiazol-2-yl]-hydrazine (10d)

Gray solid, Yield: 88%; M.p: 255–256°C; IR: 3250 (NH), 3050 (sp² CH), 2922 (sp³ CH), 1616 (C = N), 1491, 1371, 1272, 1208, 1172, 1094, 1010 cm^-1. ¹H NMR (850 MHz, DMSO-d₆): 2.88 (t, J = 8.5Hz, 2H, CH₂), 3.08 (t, J = 8.5 Hz, 2H, 2H₂), 7.39 (s, 1H, thiazole-H), 7.46 (t, J = 8.5 Hz, 2H, ArH), 7.52–7.55 (m, 3H, ArH), 7.60 (s, 1H, NH), 7.88–7.91 (m, 2H, ArH). ¹³C NMR (DMSO-d₆): 28.0 (CH₂), 28.4 (CH₂), 105.1, 122.8, 123.5, 127.7, 129.0, 129.1 130.6, 132.3, 137.5, 150.7, 155.4, 156.5, 165.9, 169.9. MS(m/z): 419 (M⁺ +2,100), 418 (M⁺+1,43), 417(M+, 76), 210 (67), 174 (21), 168 (19), 146 (5), 136(5), 129 (30), 115 (42), 111 (12), 102 (59), 89 (25), 75 (21), 63 (10), 51 (5). Elemental Analysis (%): (C₁₈H₁₃BrClN₃S; Mwt: 418.74). Calc. (Found) C, 51.63 (51.54); H, 3.13 (3.08); N, 10.03 (9.98)%.

N’-(5-bromo-2,3-dihydro-1H-inden-1-ylidene)-2-oxo-N’’-phenylpropanehydrazonhydrazide (Formazan, 14a)

Yellow solid, Yield: 89%; M.p: 190°C; IR: 3294, 3205 (2NH), 3025 (sp² CH), 1667 (C = O), 1600 (C = N), 1485, 1247, 1148 cm^-1. ¹H NMR(850 MHz, CDCl₃): 2.56 (s, 3H, CH₃), 2.78 (t, J = 8.5 Hz, 2H, CH₂), 3.16 (t, J = 8.5Hz, 2H, CH₂), 6.93 (t, J = 8.5Hz,1H, ArH), 7.17 (d, J = 8.5 Hz, 2H, ArH), 7.33 (t, J = 8.5Hz, 2H, ArH), 7.45–7.50 (m, 3H, ArH), 8.47 (s, 1H, NH), 11.62 (s, 1H, NH). MS(m/z): 384 (M⁺, 3), 212 (5), 210 (8), 176 (14), 115 (26), 105 (34), 102 (17), 92 (25), 77 (100), 70 (12), 65 (30). Elemental Analysis (%): (C₁₈H₁₇BrN₄O; Mwt: 385.27). Calc. (Found) C, 56.12 (56.03); H, 4.45 (4.31); N, 14.54 (14.42)%.

N’-(5-bromo-2,3-dihydro-1H-inden-1-ylidene)-N’’-(4-chlorophenyl)-2-oxopropanehydrazon-hydrazide (Formazan, 14b)

Yellow solid, Yield: 87%; M.p: 197–198°C; IR: 3331, 3318 (2NH), 1670(C = O), 1596(C = N), 1361, 1243, 1159 cm^-1. ¹H NMR(850 MHz, CDCl₃): 2.56 (s, 3H, CH₃), 2.83 (t, J = 8.5Hz, 2H, CH₂), 3.19 (t, J = 8.5Hz, 2H, CH₂), 7.10 (d, J = 8.5Hz, 2H, ArH), 7.3 (s, 1H, ArH), 7.45–7.53 (m, 4H, ArH), 8.50 (s, 1H, NH), 11.67 (s, 1H, NH). ¹³C NMR (CDCl₃): 23.5(CH₃), 25.1(CH₂), 28.2(CH₂), 114.1, 121.5, 124.2, 125.1, 128.9, 129.3, 130.7, 132.2, 136.4, 142.6, 149.2, 152.6, 192.9 (C = O). MS(m/z): 420 (M⁺+1, 3), 419 (M⁺, 4), 210 (76), 166 (8), 141 (63), 129 (45), 115 (59), 111 (100), 99 (45), 89 (16), 75 (40), 70 (27), 63 (23), 51 (10). Elemental Analysis (%): (C₁₈H₁₆BrClN₄O; Mwt: 419.71). Calc. (Found) C, 51.51 (51.42); H, 3.84 (3.75); N, 13.35 (13.29)%.

Ethyl 2-(2-(5-bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazineyl)-2-(2-phenyl-hydrazineylidene)acetate (Formazan, 15)

Red solid, Yield: 87%; M.p: 174–176°C IR: 3329, 3252 (2NH), 3050 (sp² CH), 2984 (sp³ CH), 1693 (C = O), 1602 (C = N), 1489, 1370, 1247, 1165 cm^-1. ¹H NMR(850 MHz, CDCl₃): 1.44 (t, J = 8.5Hz, 3H, CH₃), 2.78 (t, J = 8.5Hz, 2H, CH₂), 3.17 (t, J = 8.5Hz, 2H, CH₂), 4.36 (q, J = 8.5 Hz, 2H, CH₂), 6.89 (t, J = 8.5Hz, 1H, ArH), 7.15 (d, J = 8.5Hz, 2H, ArH), 7.30 (t, J = 8.5Hz, 2H, ArH), 7.48 (d, J = 8.5Hz, 2H, ArH), 7.50 (s, 1H, ArH), 8.37 (s, 1H, NH), 11.46 (s, 1H, NH). ¹³C NMR (CDCl₃): 14.2 (CH₃), 25.0 (CH₂), 28.2 (CH₂), 62.3 (CH₂), 112.9, 120.1, 121.6, 124.1, 124.6, 128.9, 129.2, 130.7, 136.4, 144.2, 149.1, 152.1, 162.6 (C = O). MS(m/z): 416 (M⁺ +2, 5), 415 (M⁺+1, 5), 414 (M⁺, 7), 206 (5), 129 (11), 115 (21), 105 (30), 92 (17), 89 (5), 77 (100), 65 (20), 51 (9). Elemental Analysis (%): (C₁₉H₁₉BrN₄O₂; Mwt: 415.29). Calc. (Found): C, 54.95 (54.81); H, 4.61 (4.53); N, 13.49 (13.29)%.

2-(2-(5-Bromo-2,3-dihydro-1H-inden-1-ylidene)hydrazineyl)-N-phenyl-2-(2-phenyl-hydrazineylidene)acetamide (Formazan, 16)

Yellow solid, Yield: 82%; M.p: 230–232°C IR: 3356, 3326, 3247 (3NH), 3052 (sp² CH), 1662 (C = O), 1602 (C = N), 1539, 1488, 1444, 1226, 1168 cm^-1. ¹H NMR(850 MHz, CDCl₃): 2.83 (m, 2H, CH₂), 3.19 (t, J = 8.5Hz, 2H, CH₂), 6.92–7.66 (m, 13H, ArH), 8.81 (s, 1H, NH), 9.13(s, 1H, NH), 11.30 (s, 1H, NH). ¹³C NMR (CDCl₃): 28.3 (CH₂), 29.7 (CH₂), 112.5 119.5, 119.9, 121.6, 124.2, 124.4, 128.9, 129.1, 129.4, 130.7, 136.4, 137.2, 144.2, 149.3, 153.1, 159.1 (C = O). MS(m/z): 463 (M⁺ +2, 1), 462 (M⁺+1, 1), 461 (M⁺,1), 147 (7), 129 (9), 120 (24), 115 (14), 107 (23), 102 (10), 92 (31), 77 (100), 65 (23), 51 (12). Elemental Analysis (%): (C₂₃H₂₀BrN₅O; Mwt: 462.35). Calc. (Found): C, 59.75 (59.63); H, 4.36 (4.29); N, 15.15 (15.05)%.

2.4. Biological studies

Anticancer screening.

A- Cell culture. The cells were obtained from the Egyptian Holding Company for Biological Products & Vaccines (VACSERA), Giza, Egypt, then maintained in the tissue culture unit. The cells were grown in RBMI‐1640 medium, supplemented with 10% heat-inactivated FBS, 50 units/mL of penicillin, and 50 mg/mL of streptomycin, and maintained at 23 in a humidified atmosphere containing 5% CO₂. The cells were maintained as monolayer culture by serial sub‐culturing. Cell culture reagents were obtained from Lonza (Basel, Switzerland). The anticancer activity of the rested compounds was evaluated against Gastric (SNU-16) Patch number (ATCC CRL-5822) (Gastric cancer) and COLO205 cells (colon cancer) [47, 49].

The sulforhodamine B (SRB) cytotoxicity assay.

Sulforhodamine B(SRB) assay method was used to evaluate cytotoxicity [50, 51]. Exponentially growing cells were collected using 0.25% Trypsin‐EDTA seeded in 96‐well plates at 1000‐2000 cells/well in RBMI‐1640 supplemented medium. After 24 h, cells were incubated for 72 h with various concentrations of the tested compounds. Following 72 h treatments, the cells would be fixed with 10% trichloroacetic acid for 1 h at 4°C. Wells were stained for 10 minutes at room temperature with 0.4% SRBC (Sulphorhodamine B) dissolved in 1% acetic acid. After that, the plates were air-dried for 24 h. At the same time, the dye was dissolved in Tris‐HCl and left for 5 min on the shaker at 1600 rpm. Each well’s optical density (OD) was measured spectrophotometrically at 564 nm; by using an ELISA microplate reader (ChroMate‐4300, FL, USA). The IC₅₀ values were calculated according to the equation for Boltzman sigmoidal concentration-response curve using the nonlinear regression fitting models (Graph Pad, Prism Version 9).

2.5. In-Silico ADME study

A-Molecular docking.

The docking analyses were characterized by the Molecular Environment (MOE) software. Chemdraw compound preparation, Chem 3d structures, Chem 3D 16 (Molecular Modeling and Analysis; Cambridge Soft Corporation) software, to dock the complexes toward the Colon cancer (PDB = 2A4L), Gastric cancer (PDB = PDB = 2BID). The crystal structure was downloaded from the PDB at www.rcsb.org. The following procedure was applied the water molecules, co-ligand, and metal ions were removed, and the final form was obtained after 3D protonation and the correction process. The MOE site finder generated the active binding sites to create the dummy sites as the binding pocket. The default docking parameters were triangle matcher for replacing the molecule and London dG for rescoring the docking scores. The higher negative values of the docking scores were presented along with 2D and 3D structures [52–54].

B-Pharmacophore and ADME studies.

A computational study of indane derivatives based on the thiazole nucleus was performed to predict ADME properties by the QikProp3.2 tool available in Schrödinger 9.0 version (USA) and Molinspiration online property calculation toolkit to get an idea of whether the compound has optimum pharmacokinetic properties to enter higher phases of the drug development process or not. Molinspiration strategy may be described as a complex balance of various molecular properties and structural features which conclude whether the appropriate molecule is related to the known drugs [55, 56].

2.6. Assessment of the safety profile

The ProTox-II software predicts different toxicity endpoints, such as acute toxicity, hepatotoxicity, carcinogenicity, and mutagenicity [50, 56]. The Pred-hERG (human Ether-a-go-go-Related Gene) software was used to assess cardiotoxicity. It depends on statistically significant and externally predictive quantitative structure-activity relationship (QSAR) models of hERG blockage closely associated with severe and potentially fatal cardiac dysrhythmia. The SDF (structure data file) and SMILES (simplified molecular-input line-entry system) strings were used throughout the generation process [45].

3. Results and discussion

3.1 Chemistry

This research article synthesized the targeted two starting compounds, thiosemicarbazone, and hydrazone of 5-Bromo-indan-1-one 3 and 4, as indicated in Scheme 1 from the condensation reaction 1 with each of thiosemicarbazone 2 and hydrazine hydrate in refluxing acidified ethanol or pure ethanol, respectively. The structure of derivatives 3 and 4 were confirmed based on their ¹H NMR, IR, Mass, and EA (elemental analysis) data. For instance, the ¹H NMR spectrum of derivatives 3 revealed all expected signals as follows: δ = 2.84, 3.15 (two CH₂), 6.44, 7.33, 8.69 (three NH), and 7.44–7.74 (three H_Ar) ppm. The appearance of three NH singlet signals in ¹H NMR of 3 is attributed to the two protons of the amino group being non-equivalent as one NH contributed to the H-bond with the C = N group. In addition, ¹³C NMR of the same compound 3 showed ten carbon signals at δ = 26.7 (CH₂), 28.3 (CH₂), 122.9, 125.7, 128.9, 130.8, 135.8, 150.4, 156.5 and 178.7 (C = S) ppm.

[Figure omitted. See PDF.]

To synthesize a new series of thiazole rings combined with indene, we subjected the thiosemicarbazone derivative 3 to react with hydrazonoyl chloride 5a-e in dioxane/Et₃N under reflux (Scheme 2). Such reaction products can be found in one of the three tautomeric forms 6–8. Based on the wavelength (λ_max) of the product in dioxane revealed for all derivatives, three absorption bands at 458–431, 331–323, and 259–241 nm (Table 1 & Fig 3). The appearance of λ_max at 458-431nm excluded the hydrazone tautomer 8 and proved the existence of one of the two azo-form 6 or 7 [32]. Double irradiation of the CH₃ group of thiazole derivative 6a or 7a at δ = 2.69 ppm in the NOE difference experiment revealed no enhancement for the NH signal, which means there is no electronic interaction between the protons of CH₃ and NH groups [57]. This result confirmed the tautomeric form 6 rather than structure 7. Recording the electronic absorption of the unsubstituted thiazole derivative 6a in different solvents with different polarities (Table 1), we noted no change in the absorption bands in three solvents (ethanol, acetone, and acetonitrile). Still, there is a strong bathochromic shift in the case of DMF and DMSO. This shift indicated the derivative 6a in two tautomers, 6a and 7a, in the high dielectric solvents DMF and DMSO.

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

Similarly, phenacyl bromide derivatives 9a-d were reacted with thiosemicarbazone derivative 3 to afford the thiazole derivatives 10 or 11, as illustrated in Scheme 3. However, spectral analysis data proved the isolated product’s structure as 10 rather than 11. This conclusion was observed from the ¹H & ¹³C NMR data (Fig 4). For instance, the ¹H NMR spectra of thiazole derivative 10d showed the singlet signal for the thiazole-H rather than the CH₂ of thiazole in isomer 11. In addition, the ¹³C NMR (Fig 4) was completely devoid of thiazole-CH₂ in the aliphatic region, but it revealed 14 carbon signals located in the aromatic region from 105 to 170 ppm.

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

Finally, conversion of the (5-Bromo-indan-1-ylidene)-hydrazine (4) to the formazan derivatives was achieved via the reaction of 4 with hydrazonoyl chloride 5a, 12, and 13 in dioxane/Et₃N under reflux for 5h (Scheme 4). As a result, the formed formazan derivatives 14–16 were assured based on all possible spectral techniques (See Experimental part).

[Figure omitted. See PDF.]

3.2. XRD analysis

The XRD of all synthesized thiosemicarbazone derivative 3, hydrazone 4, thiazole derivatives 6a-e, 10a-d, and formazan derivatives 14a, b, 15, and 16 were recorded over 10° < 2θ < 80° range to evaluate their crystallographic features (Fig 5A–5C). Only five derivatives, 3, 10a, 14a, 15, and 16, have crystalline shapes due to the appearance of sharp peaks in their XRD charts. In contrast, the other derivatives were amorphous. Debye–Scherrer equation [58] was used to calculate the size of the crystals of the five derivatives 3, 10a, 14a, 15, and 16, and the date was listed in Table 2. The results indicated that all five derivatives were synthesized on the nanometre scale.

[Figure omitted. See PDF.]

a. 4b. The XRD chart for compound 10a. 4c. The XRD chart for compound 16.

[Figure omitted. See PDF.]

3.3 Biological studies

Anticancer screening.

The scenario of cancer disease is one of the most challenging diseases around the world significantly. Colon and gastric cancer are familiar types in the middle east. Gastric cancer is the third deadliest tumor malignancy worldwide. A new series of substituted thiazoles were tested against two cell lines, SNU-16 for gastric carcinoma and Colo205 for colon carcinoma, with two benchmarks, cis-Pt and Sunitinib. Sunitinib is an effective drug for treating both colon and gastric cancer. The mechanism of action of cisplatin has been linked to its ability to crosslink with the purine bases on the DNA, interfering with DNA repair mechanisms, causing DNA damage, and subsequently inducing apoptosis in cancer cells [44].

For SNU-16, from the results obtained in Table 3 & Fig 6, compound 10d represented an excellent IC50 result better than the two benchmarks used, while compound 16 showed IC₅₀ better than cis-platin and near Sunitinib. For Colo205, IC₅₀ values of compounds 14a and 14b are better than cis-pt values, while IC₅₀ for compounds 4, 6c, 6d, 6e, and 10a are better than the two benchmarks used. The results obtained can be explained in detail and proved by pharmacophore studies. Apoptosis is an essential phenomenon in cytotoxicity induced by anticancer drugs. It is apparent that the molecular mechanisms by which anticancer drugs induce apoptosis are mediated by death receptor-dependent and -independent pathways, which are related to the release of cytochrome c through voltage-dependent anion channels in the mitochondrial inner membrane. The release of cytochrome c is the central gate in turning on/off apoptosis. It is regulated by the interaction of proapoptotic proteins, including Bid, Bax, and Bak, and anti-apoptotic proteins, including Bcl-2 and Bcl-XL, and a specific class of inhibitors of apoptosis proteins (IAPs) including Akt, survivin, and heat-shock proteins. Drug sensitivity can be enhanced by the introduction of proapoptotic genes and the inhibition of anti-apoptotic proteins. The signal transduction pathways triggered by the central gate in mitochondria play a critical role in anticancer drug-induced apoptosis. The modulation of signal transduction pathways targeting the proteins involved in these signal transduction pathways using antisense IAPs, and growth factor antibodies may be a good strategy for enhancing the therapeutic efficacy of anticancer drugs in cancer chemotherapy [46–48].

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

The compounds show good selectivity toward cancerous cells, represented by the “selectivity index.” The selectivity index was previously used to measure selective cytotoxic activity [50]. The selectivity index (SI) can be calculated from the ratio between IC50 of the compound on a normal cell and the compound on a cancer cell line. For example, the SI of compounds 6d, 10a, 10d, and 16 are 86.98, 76.82, 74.32, and 54.13, respectively.

3.4 Computational Insilico-studies

A-Molecular docking.

Colon and gastric carcinoma are now the most widely spread types of cancer. Therefore, molecular docking studies were performed for selected compounds. According to the results obtained from biological studies, compounds 10d and 16 have been chosen to carry out molecular docking studies on gastric cancer (PDB = 2BID) and colon cancer (PDB = 2A4L). According to the literature, the two proteins were selected [46, 47, 49]. Fig 7 explains 2D and 3D snapshots of the hydrophilicity interaction to PDB = 2BID and the results obtained for Gastric cancer (PDB = 2BID) receptor. The docking score energies for compounds 10d and 16 are -5.95 and -6.446 kcal/mol, respectively (Table 4). Furthermore, compound 10d interacts from BR18 with O-LEU46(A) by hydrogen donor. Also, it interacted by S26 with OPHE173(A) by hydrogen donor, and finally by 6-ring with 6-ring PHE24(A) by pi-pi interactions (Table 5).

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

Colon cancer from biological studies, compounds 6d and 10a have chosen to carry out molecular docking studies on colon cancer (PDB = 2A4L), (Fig 8) explain 2D and 3D snapshot of the hydrophilicity interaction to PDB = 2A4L the results obtained for Colon cancer (PDB = 2A4L) receptor. The docking score energies for compounds 6d and 10a are -7.279 and -6.63 kcal/mol, respectively (Table 6). In addition, compound 6d interacts from C10 with 5-ring HIS 125 (A) by H-pi. It was also interacted by 5-ring HIS 125 (A)N CYS 118 (A) by pi-H. At the same time, compound 10a interacted from 6-ring at N-ASN132 (A) by pi-H and from 5-ring at 5-ring HIS 125 (A) by pi-pi (Table 7).

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

B-Pharmacophore and ADME studies.

The predicted pharmacokinetic/Molinspiration properties of the new starting thiosemicarbazone, hydrazone, thiazole, and formazan derivatives 3–16, are given in Tables 8 & 9. With the help of Molinspiration virtual screening, most of the tested compounds showed good bioactivity as indicated from docking studies in Tables 4–7, which indicates the drug-likeness properties against kinase inhibitor, protease, and enzyme inhibitors.

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

The Calculated distribution of activity scores (version 2011.06) for GPCR ligands, kinase inhibitors, ion channel modulators, nuclear receptor ligands, protease inhibitors, and other enzyme targets compared with scores for about 100’000 average drug-like molecules. The score allows efficient separation of active and inactive molecules [52–57].

C-1- Pred-hERG. Chemically similar compounds often bind biologically diverse protein targets, and protein structures do not always recognize identical ligands. Pharmacological and off-target relationships among proteins and a ligand set similarity help to improve the machine learning confidence by interpolating the output prediction equalized by the compound similarity criteria. This pipeline help to improve the predictions of off-target drug effects, reducing the false-negative error. The chemical similarity is one of the essential concepts in cheminformatics. One commonly used to calculate these similarity algorithm measures is the 2D Tanimoto algorithm. The resulting Tanimoto coefficient is fingerprint-based, encoding each molecule to a fingerprint "bit" position (MACCS), with each bit recording the presence ("1") or absence ("0") of a fragment of the molecule (Figs 9–13).

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

Probability map of HERG of 10d, 16, 6d, and 10a. The more depth lines and the more intense green color imply a higher positive contribution of an atom or a fragment to the hERG blockage, while pink means that it contributes to decreasing hERG blockage and gray means no contribution.

C-2. Pro-ToxII. The ProTox-II (Fig 14 & Table 10) showed that the four compounds are predicted to have oral LD₅₀ values ranging from 335 to 3500 mg/kg in a rat model with (1 s,4 s)-Eucalyptol bearing the highest values and quercetin holding the lowest one [57, 58].

[Figure omitted. See PDF.]

[Figure omitted. See PDF.]

4. Conclusion

Herein, the new series of thiazole and formazan linked to 5-Bromo-indan and their structures were assured based on all possible analytical techniques. Five derivatives were precipitated on a nanosized scale. The anticancer activity of the tested derivatives indicated that one derivative, 10d showed activity more than the two reference drugs 49used in the case of SNU-16, while the IC50 of five derivatives was better than the two benchmarks used in the case of COLO205. All potent derivatives showed a strong fit with the active site of the two tested proteins (gastric cancer (PDB = 2BID) and colon cancer (PDB = 2A4L)) in the molecular docking study. The Pharmacophore and ADME studies of the new derivatives showed that most derivatives revealed promising bioactivity, which indicates the drug-likeness properties against kinase inhibitors, protease, and enzyme inhibitors. In addition, the ProTox-II showed that the four compounds 10d, 16, 6d, and 10a are predicted to have oral LD₅₀ values ranging from 335 to 3500 mg/kg in a rat model with (1 s,4 s)-Eucalyptol bearing the highest values and quercetin holding the lowest one. Pred-hERG model played an important rule as SAR tool with predication of the most common like drugs like the new series synthesized with the interpretation of Probability of toxicity for compounds.

Supporting information

S1 File. All spectral charts and the pictures of instruments were listed in the suplementry file.

https://doi.org/10.1371/journal.pone.0274459.s001

(DOCX)

Citation: Alfaifi GH, Farghaly TA, Magda H. Abdellattif (2023) Indenyl-thiazole and indenyl-formazan derivatives: Synthesis, anticancer screening studies, molecular-docking, and pharmacokinetic/ molin-spiration properties. PLoS ONE 18(3): e0274459. https://doi.org/10.1371/journal.pone.0274459

About the Authors:

Ghaidaa H. Alfaifi

Roles: Conceptualization, Data curation, Formal analysis, Methodology

Affiliation: Chemistry Department, Faculty of Applied Sciences, Umm Al-Qura University, Makkah, Saudi Arabia

Thoraya A. Farghaly

Roles: Data curation, Investigation, Project administration, Writing – review & editing

E-mail: [email protected], [email protected]

Affiliation: Chemistry Department, Faculty of Applied Sciences, Umm Al-Qura University, Makkah, Saudi Arabia

Magda H. Abdellattif

Roles: Funding acquisition, Software, Validation, Visualization, Writing – original draft

Affiliation: Department of Chemistry, College of Science, Taif University, Taif, Saudi Arabia

ORICD: https://orcid.org/0000-0002-8562-4749

References

1. Lagergren P, Schandl A, Aaronson NK et al. Cancer survivorship: an integral part of Europe’s research agenda. Mol. Oncol. 13(3), 624–635 (2018).

2. (a) M. S. Malik, R. A. Alsantali, A. Y. A. Alzahrani, Q. M. S. Jamal, E. M. Hussein, K. A. Alfaidi, M. M. Al-Rooqi, R. J. Obaid, M. A. Alsharif, S. F. Adil, R. S. Jassas, Z. Moussa, S. A. Ahmed. Multicomponent synthesis, cytotoxicity, and computational studies of novel imidazopyridazine-based N-phenylbenzamides. Journal of Saudi Chemical Society, 2022, 26, 101449; (b) M. S. Malik, R. I. Alsantali, Q. M. S. Jamal, Z. S. Seddigi, M. Morad, M. A. Alsharif, E. M. Hussein, R. S. Jassas, M. M. Al-Rooqi, Z. Abduljaleel, A. O. Babalgith, H. M. Altass, Z. Moussa, S. A. Ahmed. New Imidazole-Based N-Phenylbenzamide Derivatives as Potential Anticancer Agents: Key Computational Insights. Front. Chem., 2022, 9, 808556. https://doi.org/10.3389/fchem.2021.808556

3. (a) O. A. Azher, A. Hossan, R. A. Pashameah, A. Alsoliemy, A. Alharbi, T. M. Habeebullah, N. M. El-Metwaly. Synthesis, anticancer evaluation, and molecular modeling study of new 2-(phenylamino)pyrazolo[1,5-a]pyrimidine analogues. Arabian Journal of Chemistry, 2023, 16, 104437; (b) P. Yadav, A. Kumar, I. Althagafi, V. Nemaysh, R. Rai, R. Pratap. The Recent Development of Tetrahydro-Quinoline/Isoquinoline Based Compounds as Anticancer Agents Current Topics in Medicinal Chemistry, 2021, 21, 1587-1622; (c) R. Shah, T. M. Habeebullah, F. Saad, I. Althagafi, A. Y. Al-dawood, A. M. Al-Solimy, Z. A. Al-Ahmed, F. Al-Zahrani, T. A. Farghaly, N. El-Metwaly. Characterization of new Co(II) complexes and photographic monitoring for their toxic impact on breast cancer cells according to simulation study. Applied Organometallic Chemistry, 2020, 34, e5886

4. Awasthi R, Roseblade A, Hansbro PM, Rathbone MJ, Dua K, Bebawy M. Nanoparticles in cancer treatment: opportunities and obstacles. Curr. Drug Targets 19(1), 1696–1709 (2018). pmid:29577855

5. Shaaban M. R., Farghaly T. A., Alsaedi A. M. R., Abdulwahab H. G. Microwaves assisted synthesis of antitumor agents of novel azoles, azines, and azoloazines pendant to phenyl sulfone moiety and molecular docking for VEGFR-2 kinase. Journal of Molecular Structure 1249 (2022) 131657

6. Almehmadi S. J., Alsaedi A. M.R., Harras M. F., Farghaly T. A. Synthesis of a new series of pyrazolo[1,5-a]pyrimidines as CDK2 inhibitors and anti-leukemia. Bioorganic Chemistry 117 (2021) 105431.

7. Mahmoud H. K., Gomha S. M., Farghaly T. A., Awad H. M. Synthesis of thiazole Linked imidazo[2,1-b]thiazoles as Anticancer agents. Polycyclic aromatic compounds, 2021, VOL. 41, NO. 8, 1608–1622.

8. Alsaedi A. M. R., Almehmadi S. J., Farghaly T. A., Harras M. F., Khalil K. D. VEGFR2 and hepatocellular carcinoma inhibitory activities of trisubstituted triazole derivatives Journal of Molecular Structure 1250 (2022) 131832.

9. Sengel-Turk C. T., Gumustas M., Uslu B., Ozkan S. A. Chapter 10 - Nanosized Drug Carriers for Oral Delivery of Anticancer Compounds and the Importance of the Chromatographic Techniques. Nano- and Microscale Drug Delivery Systems Design and Fabrication, 2017, 165–195.

10. Hemalatha K., Madhumitha G., Kajbafvala A., Anupama N., Sompalle R., Roopan S. M. Function of Nanocatalyst in Chemistry of Organic Compounds Revolution: An Overview. Journal of Nanomaterials, 2013, ID 341015 | https://doi.org/10.1155/2013/341015

11. Alsaedi A. M. R., Farghaly T. A., Shaaban M. R. Fluorinated azole anticancer drugs: Synthesis, elaborated structure elucidation and docking studies Arabian J. Chem. (2022) 15, 103782.

12. Althagafi I. I., Abouzied A. S., Farghaly T. A., Al-Qurashi N. T., Alfaifi M. Y., Shaaban M. R., et al. Novel Nano-sized bis-indoline Derivatives as Antitumor Agents 391. J. Heterocyclic Chem., 56, 391–399 (2019).

13. Ayati A, Emami S, Asadipour A, Shafiee A, Foroumadi A. Recent applications of 1,3-thiazole core structure in the identification of new lead compounds and drug discovery. Eur. J. Med. Chem. 97, 699–718 (2015). pmid:25934508

14. Keating GM. Dasatinib: a review in chronic myeloid leukaemia and Ph+ acute lymphoblastic leukaemia. Drugs 77(1), 85–96 (2017). pmid:28032244

15. Lino CI, Gonc¸alves de Souza I, Borelli BM et al. Synthesis, molecular modeling studies and evaluation of antifungal activity of a novel series of thiazole derivatives. Eur. J. Med. Chem. 151, 248–260 (2018). pmid:29626797

16. Osman H, Yusufzai SK, Khan MS et al. New thiazolyl-coumarin hybrids: design, synthesis, characterization, x-ray crystal structure, antibacterial and antiviral evaluation. J. Mol. Struct. 1166, 147–154 (2018).

17. Djukic M, Fesatidou M, Xenikakis I et al. In vitro antioxidant activity of thiazolidinone derivatives of 1,3-thiazole and 1,3,4-thiadiazole. Chem. Biol. Interact. 286, 119–131 (2018).

18. Bondock S, Naser T, Ammar YA. Synthesis of some new 2-(3-pyridyl)-4,5-disubstituted thiazoles as potent antimicrobial agents. Eur. J. Med. Chem. 62, 270–279 (2013). pmid:23357308

19. Ahangar N, Ayati A, Alipour E, Pashapour A, Foroumadi A, Emami S. 1-[(2-arylthiazol-4-yl) methyl] azoles as a new class of anticonvulsants: design, synthesis, in vivo screening, and in silico drug-like properties. Chem. Biol. Drug Des. 78(5), 844–852 (2011).

20. Sharma RN, Xavier FP, Vasu KK, Chaturvedi SC, Pancholi SS. Synthesis of 4-benzyl-1,3-thiazole derivatives as potential anti-inflammatory agents: an analogue-based drug design approach. J. Enzyme Inhib. Med. Chem. 24(3), 890–897 (2009). pmid:19469712

21. Mishra CB, Kumari S, Tiwari M. Thiazole: a promising heterocycle for the development of potent CNS active agents. Eur. J. Med. Chem. 92, 1–34 (2015). pmid:25544146

22. Shawali A. S., Samy N. A. Functionalized formazans: A review on recent progress in their pharmacological activities. Journal of Advanced Research (2015) 6, 241–254. pmid:26257923

23. YAĞLIOĞLU AŞ, ŞENÖZ H. Synthesis of novel 5-substituted phenyl-3-(p-isopropylphenyl)-1-phenylformazan and their biological activities. Turkish Journal of Chemistry. 2017 Dec 20;41(6):883–891.

24. Khanna R, Palit G, Srivastava VK, Shanker K. Newer heterocycles of phenothiazine and their antiparkinsonian activity. Indian J Chem 1990;29B:556–60.

25. Misra VS, Dhar S, Chowdhary BL. Synthesis of some newer formazans and tetrazolium salts as antiviral agents. Pharmazie 1978;33(12):790–2. pmid:746062

26. Tandel DC, Desai CM, Patel D, Naik B, Matjad S. Synthesis of quinolinyldiarylformazans as antitubercular/antibacterial agents. Oriental J Chem 2001;17(3):529–30.

27. Bhosale JD, Shirolkar AR, Pete UD, Zade CM, Mahajan DP, Hadole CD, et al. Synthesis, characterization and biological activities of novel substituted formazans of 3,4-dimethyl-1Hpyrrole-2-carbohydrazide derivatives. J Pharm Res 2013;7(7):582–7.

28. Kalsi R, Pande K, Bhalla TN, Parmar SS, Barthwal JP. Novel formazans as potent anti-inflammatory and analgesic agents. Pharmacology 1988;37:218–24. pmid:3264073

29. Bhardwaj SD, Jolly VS. Synthesis, anti-HIV and anticancer activities of some new formazans. Asian J Chem 1997;9:48–51.

30. Khattab T, Haggag KM. Synthesis and spectral properties of symmetrical and asymmetrical 3-cyano-1, 5-diarylformazan dyestuffs for dyeing polyester fabrics. Egyptian Journal of Chemistry. 2017 Dec 1;60.

31. Farghaly T. A., Abbas E. M. H., Al-Soliemy A. M., Sabour R., and Shaaban M. R., Novel sulfonyl thiazolyl-hydrazone derivatives as EGFR inhibitors: Design, synthesis, biological evaluation and molecular docking studies, Bioorganic Chemistry 121 (2022): 105684. pmid:35183860

32. Alsharekh M.M., Althagafi I.I., Shaaban M.R. et al. Microwave-assisted and thermal synthesis of nanosized thiazolyl-phenothiazine derivatives and their biological activities. Res Chem Intermed 45, 127–154 (2019). https://doi.org/10.1007/s11164-018-3594-7

33. Alhasani M. A., Farghaly T. A., and El-Ghamry H. A., Mono and bimetallic complexes of pyrazolone based ligand: Synthesis, characterization, antitumor and molecular docking studies, Journal of Molecular Structure 1249 (2022): 131607.

34. Al-Soliemy A. M., Sabour R., and Farghaly T. A. Pyrazoles and Fused Pyrimidines: Synthesis, Structure Elucidation, Antitubercular Activity and Molecular Docking Study, Medicinal Chemistry 18 (2022): 181–198. pmid:33761862

35. Almehmadi S. J., Alsaedi A. M. R., Harras M. F., and Farghaly T. A. Synthesis of a new series of pyrazolo[1,5-a]pyrimidines as CDK2 inhibitors and anti-leukemia, Bioorganic Chemistry 117 (2021): 105431.

36. Abbas E. M. H., Gomha S. M., Farghaly T. A., Abdalla M. M. Synthesis of new thiazole derivatives as antitumor agents. Current Organic Synthesis, 2016, 13(3), 456–465.

37. Gomha S. M., Farghaly T. A., Alqurashi N. T., Abdou H. Y., Mousa E. K. Synthesis, Molecular docking and anticancer evaluation of new arylazothiazoles. Current of Organic Synthesis, 2017, 14, 620–631.

38. Santana T. I., Barbosa M. O., Gomes P. A. T. M., Cruz A. C. N., Silva T. G., Leite A. C. L. Synthesis, anticancer activity and mechanism of action of new thiazole derivatives. European Journal of Medicinal Chemistry, 2018, 144, 874–886. pmid:29329071

39. Shaaban M. R., Farghaly T. A., Alsaedi A. M. R. Synthesis, Antimicrobial and Anticancer Evaluations of Novel Thiazoles Incorporated Diphenyl Sulfone Moiety. Polycyclic Aromatic Compounds, 2022, inpress. https://doi.org/10.1080/10406638.2020.1837887

40. Farghaly T. A., Masaret G. S., Muhammad Z. A., Harras M. F. Discovery of thiazole-based-chalcones and 4-hetarylthiazoles as potent anticancer agents: Synthesis, docking study and anticancer activity. Bioorganic Chemistry, 2020, 98, 103761. pmid:32200332

41. Morigi Rita, Locatelli Alessandra, Leoni Alberto and Rambaldi Mirella, Recent Patents on Thiazole Derivatives Endowed with Antitumor Activity, Recent Patents on Anti-Cancer Drug Discovery, Volume 10, Issue 3, 2015, Page: [280–297], pmid:26152151

42. Prabodh ChanderSharma Kushal KumarBansal, Archana Sharma, Diksha Sharma, Aakash Deep, Thiazole-containing compounds as therapeutic targets for cancer therapy, European Journal of Medicinal Chemistry, Volume 188, 15 February 2020, 112016, pmid:31926469

43. Altıntop M.D.; Sever B.; Akalın Çiftçi G.; Özdemir A. Design, Synthesis, and Evaluation of a New Series of Thiazole-Based Anticancer Agents as Potent Akt Inhibitors. Molecules 2018, 23, 1318. pmid:29857484

44. Dasari Shaloam, Bernard Paul, Tchounwou, Cisplatin in cancer therapy: Molecular mechanisms of action, European Journal of Pharmacology, Volume 740, 5 October 2014, Pages 364–378, https://doi.org/10.1016/j.ejphar.2014.07.025

45. Magda H. Abdellattif, Hossam Nada, and Ahmed Elkamhawy, Synthesis, Biological Evaluation, and In Silico Studies of New Heterocycles Incorporating 4,5,6,7-Tetrabromophthalimide Moiety as Potential Antibacterial and Anticancer Agents https://doi.org/10.3987/COM-21-14535

46. Kim R., Tanabe K., Uchida Y. et al. Current status of the molecular mechanisms of anticancer drug-induced apoptosis. Cancer Chemother Pharmacol 50, 343–352 (2002). https://doi.org/10.1007/s00280-002-0522-7

47. Guzmán L., Villalón K., Marchant M.J. et al. In vitro evaluation and molecular docking of QS-21 and quillaic acid from Quillaja saponaria Molina as gastric cancer agents. Sci Rep 10, 10534 (2020). https://doi.org/10.1038/s41598-020-67442-3

48. Abdellattif MH, Abdel-Rahman AAH, Arief MMH, Mouneir SM, Ali A, Hussien MA, et al. (2021) Novel 2-Hydroselenonicotinonitriles and Selenopheno[2, 3-b]pyridines: Efficient Synthesis, Molecular Docking-DFT Modeling and Antimicrobial Assessment. Front. Chem. 9:672503. pmid:34041224

49. Govindarasu Mydhili, Ganeshan Shalini, Ansari Mohammad Azam, Alomary Mohammad N., AlYahya Sami, Alghamdi Saad, et al, In silico modeling and molecular docking insights of kaempferitrin for colon cancer-related molecular targets, Journal of Saudi Chemical Society (2021) 25, 101319, https://doi.org/10.1016/j.jscs.2021.101319

50. Razak S., Afsar T., Bibi N. et al. Molecular docking, pharmacokinetic studies, and in vivo pharmacological study of indole derivative 2-(5-methoxy-2-methyl-1H-indole-3-yl)-N′-[(E)-(3-nitrophenyl) methylidene] acetohydrazide as a promising chemoprotective agent against cisplatin induced organ damage. Sci Rep 11, 6245 (2021). pmid:33737575

51. Liebowitz L. D.; Ashbee H. R.; Evans E. G. V.; Chong Y.; Mallatova N.; Zaidi M.; et al. Diagn. Microbiol. Infect. Dis. 2002, 4, 27‐33.

52. Howsaui H.B.; Basaleh A.S.; Abdellattif M.H.; Hassan W.M.I.; Hussien M.A. Synthesis, Structural Investigations, Molecular Docking, and Anticancer Activity of Some Novel Schiff Bases and Their Uranyl Complexes. Biomolecules 2021, 11, 1138. pmid:34439805

53. Lobinski R., Edmonds J., Suzuki K., and Uden P. (2000). Species-selective determination of selenium compounds in biological materials. Pure Appl. Chem. 72, 447–462.

54. Lipinski C. A., Lombardo F., Dominy B. W., and Feeney P. J. (1997) Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv. Drug Deliv. Rev. 23, 3–25.

55. Murad H.A.S., Alqurashi T.M.A. & Hussien M.A. Interactions of selected cardiovascular active natural compounds with CXCR4 and CXCR7 receptors: a molecular docking, molecular dynamics, and pharmacokinetic/toxicity prediction study. BMC Complement Med Ther 22, 35 (2022). pmid:35120520

56. Elkamhawy A.; Ammar U.M.; Paik S.; Abdellattif M.H.; Elsherbeny M.H.; Lee K.; et al. Scaffold Repurposing of In-HouseSmall Molecule Candidates Leads to Discovery of First-in-Class CDK-1/HER-2 Dual Inhibitors: In Vitro and In Silico Screening. Molecules 2021, 26, 5324. pmid:34500757

57. Farghaly T. A., Abdallah M. A., Mahmoud H. K. Regio- and Site Selectivity in the Reactions of Hydrazonoyl Chlorides with 3-substituted Indolin-2-one Derivatives. J. Heterocycl. Chem., 2017, 54, 1450–1456.

58. Cullity B. D.; Elements of X-ray Diffraction, second ed.Addison-Wesley, Massachusetts, 1978.