Selective inhibition of carbonic anhydrase IX and XII by coumarin and psoralen derivatives

Figures & data

Table 1. Chemical, analytical, and physical data of derivatives EMAC10157 a-b-d-g and EMAC10160 a-b-d-g.

Compound	R	C, %; H, %		M.P., °C	Yield, %^a	Aspect
		Calc.	Found
EMAC10157a	4-CH3	C, 71.07; H, 6.20	C, 71.10; H, 6.19	132–133	80	White crystals
EMAC10157b	4-OCH3	C, 68.48; H, 5.98	C, 68.74; H, 5.97	127–130	89	Pale brown crystals
EMAC10157d	4-F	C, 67.60; H, 5.44	C, 67.79; H, 5.45	164–166	90	White crystals
EMAC10157g	4-C6H5	C, 74.36; H, 5.82	C, 74.55; H, 5.45	138–140	71	White crystals
EMAC10160a	4-CH3	C, 74.24; H, 5.98	C, 74.19; H, 5.99	226–229	98	Grey crystals
EMAC10160b	4-OCH3	C, 71.41; H, 5.75	C, 71.68, H, 5.76	202–205	97	Green crystals
EMAC10160d	4-F	C, 70.58; H, 5.18	C, 70.30; H, 5.16	135–139	92	Grey crystal
EMAC10160g	4-C6H5	C, 77.24; H, 5.62	C, 77.52; H, 5.60	248–250	97	Pale brown crystal

a Yields are referred to the last step of the synthetic pathway.

Table 2. ¹H NMR and ¹³C NMR data of derivatives EMAC10157a-b-d-g and EMAC10160a-b-d-g.

Compound	^1H NMR and ^13C NMR δ (ppm)
EMAC10157a	1H NMR (400 MHz, DMSO) δ 7.93 (d, 2H, J = 8), 7.61 (d, 1H, J = 9.2), 7.39 (d, 2H, J = 8), 7.00 (d, 1H, J = 8.8), 5.74 (s, 2H), 3.68 (s, 2H), 3.62 (s, 3H), 2.83–2.79 (m, 2H), 2.41 (s, 3H), 2.37 (s, 3H), 1.64–1.55 (m, 2H), 0.95–0.91 (t, 3H). 13C NMR (100 MHz, DMSO) δ 193.70, 170.64, 160.88, 158.24, 150.61, 149.69, 144.37, 131.78, 129.34 (2C), 127.96 (2C), 123.92, 116.97, 115.87, 113.65, 108.77, 70.55, 51.82, 32.37, 24.34, 21.70, 21.22, 15.10, 13.96.
EMAC10157b	1H NMR (400 MHz, DMSO) δ 8.01 (d, 2H, J = 8.8), 7.61 (d, 1H, J = 8), 7.09 (d, 2H, J = 8), 6.98 (d, 1H, J = 8), 5.70 (s, 2H), 3.86 (s, 3H), 3.68 (s, 2H), 3.62, (s, 3H), 2.82–2.79 (m, 2H), 2.36 (s, 3H), 1.64–1.55 (m, 2H), 0.95–0.91 (t, 3H) 13C NMR (100 MHz, DMSO) δ 192.48, 170.64, 163.61, 160.89, 158.29, 150.60, 149.69, 130.23 (2C), 127.15 123.90, 116.96, 115.84, 114.05 (2C), 113.61, 108.76, 70.35, 55.60, 51.81, 32.36, 24.34, 21.70, 15.09, 13.96.
EMAC10157d	1H NMR (400 MHz, DMSO) δ 8.13–8.10 (m, 2H), 7.62 (d, 1H, J = 9.2), 7.45–7.40 (m, 2H), 7.03 (d, 1H, J = 9.2), 5.77 (s, 2H), 3.68 (s, 2H), 3.62 (s, 3H), 2.83–2.79 (m, 2H), 2.37 (s, 3H), 1.64–1.55 (m, 2H), 0.95–0.91 (t, 3H). 13C NMR (100 MHz, DMSO) δ 192.90, 170.65, 166.59, 164.08, 160.89, 158.15, 150.60, 149.70, 131.01 (2C), 123.94, 116.98, 116.01, 115.91 (2C), 113.70, 108.79, 70.55, 51.82, 32.36, 24.33, 21.70, 15.10, 13.95.
EMAC10157g	1H NMR (400 MHz, DMSO) δ 8.12 (d, 2H, J = 8.4), 7.88 (d, 2H, J = 8.4), 7.78 (d, 2H, J = 7.2), 7.65–7.61 (m, 1H), 7.53 (t, 2H, J = 7.2), 7.45 (t, 1H, J = 7.2), 7.06–7.02 (m, 1H), 5.82 (s, 2H), 3.68 (s, 2H), 3.62 (s, 3H), 2.85–2.81 (m, 2H),2.37 (s, 3H), 1.64–1.58 (m, 2H), 0.96–0.92 (t, 3H). 13C NMR (100 MHz, DMSO) δ 193.78, 170.63, 160.88, 158.22, 150.62, 149.68, 145.17, 138.77, 133.07, 129.10 (2C), 128.61 (2C), 127.02 (2C), 126.96 (2C), 123.94, 116.99, 115.90, 113.68, 108.81, 73.94, 70.69, 51.81, 32.37, 24.37, 21.73, 15.11, 13.98.
EMAC10160a	1H NMR (400 MHz, DMSO) δ 12.47 (bs, 1H), 8.41 (s, 1H), 8.03 (s, 1H), 7.69 (d, 2H, J = 8), 7.34 (d, 2H, J = 7.6),3.65 (s, 2H), 3.05–3.01 (m, 2H), 2.51 (s, 3H), 2.38 (s, 3H), 1.78–1.69 (m, 2H), 0.97–0.94 (t, 3H). 13C NMR (100 MHz, DMSO) δ 171.49, 160.72, 154.93, 149.65, 147.36, 143.62, 137.11, 129.72 (2C), 127.82, 127.11 (2C), 122.21, 121.45, 117.83, 116.71, 114.20, 112.93, 32.90, 24.85, 21.93, 20.79, 15.64, 13.83.
EMAC10160b	1H NMR (400 MHz, DMSO) δ 12.47 (bs, 1H), 8.37 (s, 1H), 8.01 (s,1H), 7.73 (d, 2H, J = 8), 7.10 (d, 2H, J = 8.8), 3.82 (s, 3H), 3.65 (s, 2H), 3.04–3.01 (m, 2H), 2.51 (s, 3H), 1.71–1.68 (m, 2H), 0.97–0.94 (t, 3H). 13C NMR (100 MHz, DMSO) δ 171.50, 160.73, 158.92, 154.89, 149.66, 147.34, 143.17, 128.47 (2C), 123.00, 122.31, 121.18, 117.79, 116.66, 114.62 (2C), 114.16, 112.89, 55.17, 32.90, 24.84, 21.92, 15.65, 13.82.
EMAC10160d	1H NMR (400 MHz, DMSO) δ 12.47 (bs, 1H), 8.46 (s, 1H), 8.03 (s, 1H), 7.87–7.85 (m, 2H), 7.39–7.35 (m, 2H), 3.65 (s, 2H), 3.05–3.01 (m, 2H), 2.52 (s, 3H), 1.78–1.69 (m, 2H), 0.97–0.94 (t, 3H). 13C NMR (100 MHz, DMSO) δ 171.48, 162.91, 160.68, 160.51, 154.88, 149.67, 147.42, 129.23, 129.23, 127.21, 121.97, 120.59, 117.90, 116.81, 116.16, 115.94, 114.16, 112.98, 32.90, 27.45, 24.84, 21.92, 15.67, 13.82.
EMAC10160g	1H NMR (400 MHz, DMSO) δ 12.49 (bs, 1H), 8.54 (s, 1H), 8.13 (s, 1H), 7.92 (d, 2H, J = 8), 7.84 (d, 2H, J = 8.4), 7.75 (d, 2H, J = 7.2), 7.51 (t, 2H, J = 7.2), 7.40 (t, 1H, J = 7.2), 3.67 (s, 2H), 3.07–3.04 (m, 2H), 2.55 (s, 3H), 1.80–1.71 (m, 2H), 0.99–0.95 (t, 3H). 13C NMR (100 MHz, DMSO) δ 171.49, 160.71, 155.01, 149.69, 147.44, 144.21, 139.56, 139.42, 129.93, 129.01 (2C), 127.73 (2C), 127.59, 127.37 (2C), 126.54 (2C), 122.04, 121.13, 117.92, 116.84, 114.35, 113,03, 32.92, 24.88, 21.94, 15.71, 13.85.

Figure 1. Newly synthesised coumarin and psoralen derivatives.

Figure 2. (a) Esterase activity of carbonic anhydrase on compound EMAC10157g³⁴. (b, c) Oral bioavailability radar profile.

Scheme 1. Synthetic pathway to compounds EMAC10157 a-b-d-g and EMAC10160 a-b-d-g. Reagents and conditions: (i) dimethylacetylsuccinate, H₂SO₄ 98% R.T.; (ii) α-halogeno arylketone, dry acetone, K₂CO₃, reflux; (iii) NaOH 1 N, reflux.

Table 3. Inhibition data towards hCA I, II, IX, and XII of compounds EMAC10157a-b-d-g and EMAC10160 a-b-d-g.

Compound	R	Ki (nM)*
		hCA I	hCA II	hCA IX	hCA XII
EMAC10157a	4-CH3	>10,000	>10,000	247.7	350.3
EMAC10157b	4-OCH3	>10,000	>10,000	352.7	324.2
EMAC10157d	4-F	>10,000	>10,000	239.5	257.1
EMAC10157g	4-C6H5	>10,000	>10,000	135.2	283.1
EMAC10160a	4-CH3	>10,000	>10,000	467.3	758.1
EMAC10160b	4-OCH3	>10,000	>10,000	489.3	859.4
EMAC10160d	4-F	>10,000	>10,000	379.7	460.0
EMAC10160g	4-C6H5	>10,000	>10,000	397.7	550.0
AAZ		250	12	25	5.7

* Standard error (from three different assays) were in the range of plus/minus 10% of the reported values.

Figure 3. 3D representation of the putative binding mode obtained by docking experiments. (a) hCA-IX – EMAC10157g, (b) hCA-IX – EMAC10157g-openE, and (c) hCA-IX – EMAC10157g-openZ and the relative 2D representation of the complexes stabilising interactions with the binding site residues represented with different colour depending on their chemical-physical properties: green, hydrophobic; cyan, polar; violet, positive; red, negative charged residues; grey, metal atoms. Instead, magenta arrows indicate the formation of a hydrogen bond between protein and ligand, while grey lines indicate the interaction with the complexed ion.

Figure 4. 3D representation of the putative binding mode obtained by docking experiments. (a) hCA-XII – EMAC10157g, (b) hCA-XII – EMAC10157g-openE, and (c) CA-XII- EMAC10157g-openZ and the relative 2D representation of the complexes stabilising interactions with the binding site residues.

Supuran CT. Coumarin carbonic anhydrase inhibitors from natural sources. J Enzyme Inhibition Med Chem 2020;35:1462–70.

 PubMed Web of Science ®Google Scholar