2-Thiopyrimidine/chalcone hybrids: design, synthesis, ADMET prediction, and anticancer evaluation as STAT3/STAT5a inhibitors

Figures & data

Figure 1. The designed strategy for 2-TP/chalcone hybrids as new anticancer STAT inhibitors.

Scheme 1. Synthesis of 2-thiopyrimidine derivatives 4a–f.

Scheme 2. Synthesis of chloroacetyl aminochalcone derivatives 8a–c.

Scheme 3. Synthesis of the target compounds 9a–r.

Table 1. Cytotoxicity results of pyrimidine/chalcone hybrids 9a–r against three different cancer cell lines.

Compound	R	R^1	R^2	R^3	(IC50 μM)±SD
					K-562	MCF-7	HT-29
9a	OCH3	CH3	H	H	18.40 ± 0.76	3.56 ± 0.14	2.20 ± 0.04
9b	OCH3	CH3	CH3	H	3.62 ± 0.08	22.45 ± 1.57	16.36 ± 0.76
9c	OCH3	CH3	OCH3	OCH3	11.42 ± 0.04	4.25 ± 0.13	8.70 ± 0.28
9d	OCH3	NO2	H	H	0.77 ± 0.03	14.16 ± 0.74	25.92 ± 1.67
9e	OCH3	NO2	OCH3	H	7.05 ± 0.28	18.74 ± 0.92	7.31 ± 0.36
9f	OCH3	NO2	OCH3	OCH3	1.37 ± 0.03	5.77 ± 0.22	9.41 ± 0.43
9g	OCH3	OCH2CH2Cl	H	H	3.17 ± 0.11	10.40 ± 0.64	10.55 ± 0.47
9h	OCH3	OCH2CH2Cl	OCH3	H	7.07 ± 0.31	7.70 ± 0.32	18.77 ± 0.88
9i	OCH3	OCH2CH2Cl	OCH3	OCH3	9.71 ± 0.41	11.47 ± 0.81	11.47 ± 0.23
9j	Cl	CH3	H	H	4.26 ± 0.06	6.26 ± 0.24	6.26 ± 0.34
9k	Cl	CH3	OCH3	H	9.95 ± 0.29	28.65 ± 1.39	5.61 ± 0.18
9l	Cl	CH3	OCH3	OCH3	42.60 ± 1.99	13.46 ± 0.62	2.37 ± 0.07
9m	Cl	NO2	H	H	3.86 ± 0.14	3.90 ± 0.09	7.90 ± 0.22
9n	Cl	NO2	OCH3	H	1.05 ± 0.02	17.36 ± 0.75	2.10 ± 0.06
9o	Cl	NO2	OCH3	OCH3	12.35 ± 0.72	3.62 ± 0.084	3.62 ± 0.08
9p	Cl	OCH2CH2Cl	H	H	1.74 ± 0.04	11.64 ± 0.49	11.64 ± 0.63
9q	Cl	OCH2CH2Cl	OCH3	H	5.77 ± 0.16	6.81 ± 0.25	6.06 ± 0.29
9r	Cl	OCH2CH2Cl	OCH3	OCH3	10.67 ± 0.77	1.37 ± 0.07	4.74 ± 0.27
Cisplatin					2.31 ± 0.09	6.62 ± 0.29	1.12 ± 0.06
Erlotinib					9.85 ± 0.51	10.64 ± 0.58	9.20 ± 0.41

Figure 2. Cytotoxicity (IC₅₀) of the most active derivatives and cisplatin against WI38 cell line.

Table 2. STAT3 and STAT5a inhibitory activity of compounds 9a, 9d, 9f, 9n, 9r and reference drug pacritinib.

Compound/no.	Inhibition IC50 (µM)
	STAT3	STAT5a
9a/MCF7	242.53 ± 9.24	83.78 ± 3.28
9d/K562	160.01 ± 4.59	116.31 ± 4.13
9f/K562	244.74 ± 11.07	77.65 ± 2.91
9n/K562	113.31 ± 3.22	50.75 ± 1.26
9r/MCF7	148.69 ± 3.81	63.24 ± 1.57
Pacritinib/MCF7	79.47 ± 2.17	54.35 ± 1.09
Pacritinib/K562	65.49 ± 2.55	69.81 ± 1.82

Table 3. Biological properties, prediction, and drug likeness of the target compounds.

Compound	GPCR ligand	Ion channel modulator	Kinase inhibitor	Nuclear receptor ligand	Protease inhibitor	Drug likeness score
9a	−0.43	−1.03	−0.58	−0.69	−0.43	0.49
9b	−0.56	−1.27	−0.78	−0.90	−0.51	0.54
9c	−0.73	−1.55	−1.01	−1.17	−0.62	0.96
9d	−0.61	−1.25	−0.83	−0.92	−0.53	0.07
9e	−0.78	−1.53	−1.07	−1.17	−0.65	0.11
9f	−0.99	−1.86	−1.35	−1.50	−0.80	0.49
9g	−0.69	−1.44	−0.81	−1.01	−0.56	0.83
9h	−0.88	−1.75	−1.08	−1.29	−0.70	0.79
9i	−1.11	−2.09	−1.39	−1.64	−0.87	1.18
9j	−0.39	−0.94	−0.52	−0.63	−0.44	0.82
9k	−0.50	−1.16	−0.68	−0.81	−0.50	0.80
9l	−0.64	−1.41	−0.89	−1.06	−0.58	1.07
9m	−0.55	−1.13	−0.75	−0.83	−0.52	0.43
9n	−0.69	−1.39	−0.95	−1.06	−0.62	0.40
9o	−0.88	−1.69	−1.21	−1.36	−0.74	0.63
9p	−0.61	−1.31	−0.71	−0.91	−0.54	1.14
9q	−0.79	−1.60	−0.96	−1.17	−0.66	1.05
9r	−1.00	−1.92	−1.24	−1.49	−0.81	1.40

Figure 3. Drug likeness score value (1.40) for compound 9r.

Table 4. Pharmacokinetic properties assessment of the target synthesised compounds 9a–r.

Compound	Absorption				Distribution		Metabolism (CYP) and excretion
	HIA (%)	SP LogP (cm/h)	Caco2 (nm/sc)	MDCK (nm/sc)	BBB (c.brain/c.blood)	PPB %	2C19	2C9	2D6	3A4	Pgp Inh.
9a	97.50	−1.72	54.34	0.05	0.03	98.31	No	Yes	No	Yes	Inh.
9b	97.36	−1.73	54.31	0.06	0.03	99.63	No	Yes	No	Yes	Inh.
9c	97.25	−1.74	54.26	0.06	0.03	97.83	No	Yes	No	Yes	Inh.
9d	98.78	−2.26	28.65	0.04	0.49	92.53	No	Yes	No	Yes	Inh.
9e	99.22	−2.25	29.60	0.04	0.45	96.23	No	Yes	No	Yes	Inh.
9f	99.39	−2.22	30.34	0.04	0.38	96.95	No	Yes	No	Yes	Inh.
9g	97.69	−2.09	37.98	0.05	0.02	92.10	No	Yes	No	Yes	Inh.
9h	97.54	−2.03	38.68	0.05	0.02	95.16	No	Yes	No	Yes	Inh.
9i	97.42	−1.96	39.36	0.05	0.02	95.17	No	Yes	No	Yes	Inh.
9j	97.95	−1.74	51.82	0.06	0.08	90.94	No	Yes	No	Yes	Inh.
9k	97.83	−1.74	52.19	0.07	0.05	94.17	No	Yes	No	Yes	Inh.
9l	97.69	−1.73	52.52	0.07	0.04	94.05	No	Yes	No	Yes	Inh.
9m	97.49	−2.28	33.88	0.04	0.37	93.03	No	Yes	No	Yes	Inh.
9n	97.96	−2.28	35.90	0.04	0.50	91.43	No	Yes	No	Yes	Inh.
9o	98.56	−2.27	37.81	0.04	0.57	92.17	No	Yes	No	Yes	Inh.
9p	98.11	−2.14	42.99	0.05	0.05	92.90	No	Yes	No	Yes	Inh.
9q	97.99	−2.10	43.47	0.05	0.03	91.65	No	Yes	No	Yes	Inh.
9r	97.86	−2.05	43.93	0.05	0.03	91.71	No	Yes	No	Yes	Inh.

Table 5. Toxicity assessment of the target synthesised compounds 9a–r.

Compound	AMES	Carcino-Mouse	Carcino-Rat	hERG-inhibition
9a	Mutagen	Negative	Negative	Medium
9b	Non-mutagen	Negative	Negative	Medium
9c	Non-mutagen	Negative	Negative	Medium
9d	Mutagen	Negative	Negative	Medium
9e	Mutagen	Negative	Negative	Medium
9f	Mutagen	Negative	Negative	Medium
9g	Non-mutagen	Negative	Negative	Medium
9h	Non-mutagen	Negative	Negative	Medium
9i	Non-mutagen	Negative	Negative	Medium
9j	Mutagen	Negative	Negative	Medium
9k	Mutagen	Negative	Negative	Medium
9l	Non-mutagen	Negative	Negative	Medium
9m	Mutagen	Negative	Negative	Medium
9n	Mutagen	Negative	Negative	Medium
9o	Mutagen	Negative	Negative	Medium
9p	Non-mutagen	Negative	Negative	Medium
9q	Non-mutagen	Negative	Negative	Medium
9r	Non-mutagen	Negative	Negative	Medium

Figure 4. SAR study of the target compounds 9a–r.