Mono- and di-thiocarbamate inhibition studies of the δ-carbonic anhydrase TweCAδ from the marine diatom Thalassiosira weissflogii

Figures & data

Figure 1. Monothiocarbamates (MTCs) 1–15 investigated as CA inhibitors⁴.

Figure 2. Dithiocarbamates (DTCs) 16–36 investigated as CA inhibitors^1,2.

Table 1. TweCAδ, hCA I, and hCA II Inhibition Data with MTCs 1–15, DTCs 16–36, and acetazolamide (AAZ, 5-acetamido-1,3,4-thiadiazole-2-sulphonamide) as standard drug, by a stopped-flow CO₂ hydrase assay.

	RR^1NCOS^− Na^+ (1–15)		RR^1NCS2M (16–36)
			KI (nM)^a
No.	R	R^1	TweCAδ	hCA I	hCA II
1	n-Pr	n-Pr	806.7	>2000	46.7
2	Et	n-Bu	783.3	700	>2000
3	n-Bu	n-Bu	1142	909	>2000
4	i-Bu	i-Bu	>20,000	681	43.0
5	Me	CH2COOEt	>20,000	827	44.5
6		–(CH2CH2)–O–(CH2CH2)–	>20,000	569	>2000
7	H	–N(CH2CH2)N(CH3)CH2CH2–	487	>2000	35.0
8		– (CH2CH2)–NH-(CH2CH2)–	483	876	22.4
9		–(CH2CH2)-N(CH2CONHC6H11)–(CH2CH2)–	129	949	45.9
10	Me	CH2Ph	>20,000	>2000	>2000
11	H	CH2CH2Ph	997	>2000	43.7
12		HCH2CH2(3,4-di-MeO-C6H4)	67.7	891	26.7
13		– (CH2CH2)–N(3-Cl-C6H4)– (CH2CH2)–	1505	686	>2000
14		–(CH2CH2)–N(4-F-C6H4)–(CH2CH2)–	1498	895	46.8
15		–(CH2CH2)-N(4-CF3-C6H4)– (CH2CH2)–	1152	>2000	43.6
16	Me2N(CH2)2	H	8406	85.9	35.8
17	HO(CH2)3	H	8691	706	41.7
18	HO(CH2)4	H	7168	295	24.3
19	HO(CH2)5	H	8597	66.5	17.3
20		H	>20,000	494	48.7
21	(R)	H	>20,000	240	18.9
22	(S)	H	7995	615	65.9
23	–(CH2)5–	–	>20,000	252	30.1
24	–(CH2)3–CH(OH)CH2–	–	>20,000	428	60.7
25	–(CH2)4–CH(COONa)–	–	>20,000	485	80.1
26	–(CH2)3–CH(COONa)CH2–	–	8429	290	45.4
27	(R)–(CH2)3–CH(COONa)CH2–	–	93.6	496	80.5
28	(S) -(CH2)3–CH(COONa)CH2–	–	556	109	8.9
29	–(CH2)2–CH(COONa)(CH2)2–	–	8980	337	78.7
30	–(CH2)3-CH(NHAc)CH2–	–	783	910	47.9
31	–(CH2)3-CH(NHBoc)CH2–	–	9239	683	13.2
32	–CH(Me)CH2-O-(CH2)2–	–	>20,000	434	60.2
33	–CH(COONa)CH2-O– (CH2)2–	–	>20,000	84.7	78.5
34	–	–(CH2)2 N(CH2CONHC6H11)(CH2)2–	791	415	67.2
35	Ph(CH2)2	H	897	425	107
36	–	H2NO2SC6H4(CH2)2H	704	97.5	48.1
AAZ	–	–	83	250	12.1

^a Mean ± standard error (from three different assays), by a stopped-flow technique (errors were in the range of ±5–10% of the reported values).

Figure 3. Multialignment of the TweCAδ amino acid sequence with those of bacterial (HpylCA, α-CA from Helicobacter pylori, SspCA, α-CA from Sulfurihydrogenibium yellowstonensis) and human (hCA I and II) α-class enzymes. The zinc ligands of the α-CAs and the putative zinc ligands of TweCAδ are evidenced in red, whereas amino acid residues involved in the catalytic inhibition/mechanism (e.g. His64 and Asp106, hCA I numbering) are shown in green and blue, respectively.

a) Vullo D, Durante M, Di Leva FS, et al. Monothiocarbamates strongly inhibit carbonic anhydrases in vitro and possess intraocular pressure lowering activity in an animal model of glaucoma. J Med Chem 2016;59:5857–67. b) Nocentini A, Vullo D, Del Prete S, et al. Inhibition of the β-carbonic anhydrase from the dandruff-producing fungus Malassezia globosa with monothiocarbamates. J Enzyme Inhib Med Chem 2017;32:1064–70.

 PubMed Web of Science ®Google Scholar

a) Carta F, Aggarwal M, Maresca A, et al. Dithiocarbamates: a new class of carbonic anhydrase inhibitors. Crystallographic and kinetic investigations. Chem Commun (Camb) 2012;48:1868–70. b) Carta F, Aggarwal M, Maresca A, et al. Dithiocarbamates strongly inhibit carbonic anhydrases and show antiglaucoma action in vivo. J Med Chem 2012;55:1721–30. c) Monti SM, Maresca A, Viparelli F, et al. Dithiocarbamates are strong inhibitors of the beta-class fungal carbonic anhydrases from Cryptococcus neoformans, Candida albicans and Candida glabrata. Bioorg Med Chem Lett 2012;22:859–62. d) Maresca A, Carta F, Vullo D, Supuran CT. Dithiocarbamates strongly inhibit the β-class carbonic anhydrases from Mycobacterium tuberculosis. J Enzyme Inhib Med Chem 2013;28:407–11.

 PubMed Web of Science ®Google Scholar

a) Syrjänen L, Tolvanen ME, Hilvo M, et al. Characterization, bioinformatic analysis and dithiocarbamate inhibition studies of two new α-carbonic anhydrases, CAH1 and CAH2, from the fruit fly Drosophila melanogaster. Bioorg Med Chem 2013;21:1516–21. b) Winum JY, Supuran CT. Recent advances in the discovery of zinc-binding motifs for the development of carbonic anhydrase inhibitors. J Enzyme Inhib Med Chem 2015;30:321–4. c) Bozdag M, Carta F, Vullo D, et al. Dithiocarbamates with potent inhibitory activity against the Saccharomyces cerevisiae β-carbonic anhydrase. J Enzyme Inhib Med Chem 2016;31:132–6. d) Bozdag M, Carta F, Vullo D, et al. Synthesis of a new series of dithiocarbamates with effective human carbonic anhydrase inhibitory activity and antiglaucoma action. Bioorg Med Chem 2015;23:2368–76. e) Vullo D, Del Prete S, Nocentini A, et al. Dithiocarbamates effectively inhibit the β-carbonic anhydrase from the dandruff-producing fungus Malassezia globosa. Bioorg Med Chem 2017;25:1260–5. f) Aspatwar A, Hammarén M, Koskinen S, et al. β-CA-specific inhibitor dithiocarbamate Fc14-584B: a novel antimycobacterial agent with potential to treat drug-resistant tuberculosis. J Enzyme Inhib Med Chem 2017;32:832–40.

 PubMed Web of Science ®Google Scholar