Sulfonamides with Potent Inhibitory Action and Selectivity against the α‑Carbonic Anhydrase from <i>Vibrio cholerae</i>

By using <i>N</i>-α-acetyl-l-lysine or GABA scaffolds and the conversion of the terminal amino group to the guanidine one, benzenesulfonamides incorporating water solubilizing moieties were synthesized. The new compounds were medium potency inhibitors of the cytosolic carbonic anhydrase (CA, EC 4.2.1.1) isoforms I and II, and highly effective, nanomolar inhibitors of the pathogenic bacterial α-CA from <i>Vibrio cholerae</i>. These sulfonamides possess good selectivity for inhibiting the bacterial over the mammalian isoforms and may be used as tools to understand the role of bacterial CAs in pathogenesis

Benzoxaboroles as Efficient Inhibitors of the β‑Carbonic Anhydrases from Pathogenic Fungi: Activity and Modeling Study

A series of 6-substituted benzoxaboroles were investigated as inhibitors of the β-class carbonic anhydrase from three pathogenic fungi (<i>Cryptococcus neoformans</i>, <i>Candida glabrata</i>, and <i>Malassezia globosa</i>). Independently from the nature of the substituents on the phenyl of the urea/thiourea group, all reported derivatives showed nanomolar inhibitory activities against Can2 and CgNce103 vs micromolar inhibition against MgCA. Selectivity over human CA I and CA II was noticed. The observed structure–activity relationship trends have been rationalized by modeling study of selected compounds into the active site of Can2 and MgCA. The present letter demonstrates that benzoxaborole chemotype may offer interesting opportunities for the inhibition of β-CA from pathogenic fungi and for the development of antifungal agents with a new mechanism of action

Effect of CA II addition to IGL-1 solution in hepatic function.

<p>Hepatic function expressed as bile output (A) and percentage of bromosulfophthalein (BSP) excretion in bile (B) after 120 min of fatty liver normothermic <i>ex vivo</i>perfusion. CA II supplementation (IGL-1+CAII) significantly enhanced bile production and BSP clearance compared to the IGL-1 group. Ctr 2: Livers flushed and perfused <i>ex-vivo</i> without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h-normothermic <i>ex vivo</i> perfusion. IGL-1+CAII: livers preserved in IGL-1 solution (4°C, 24 h) enriched with CA II (10 ug/ml) and subjected to 2h- normothermic <i>ex vivo</i> perfusion; * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.</p

Cloning, Characterization, and Sulfonamide and Thiol Inhibition Studies of an α‑Carbonic Anhydrase from <i>Trypanosoma cruzi</i>, the Causative Agent of Chagas Disease

An α-carbonic anhydrase (CA, EC 4.2.1.1) has been identified, cloned, and characterized from the unicellular protozoan <i>Trypanosoma cruzi</i>, the causative agent of Chagas disease. The enzyme (TcCA) has a very high catalytic activity for the CO₂ hydration reaction, being similar kinetically to the human (h) isoform hCA II, although it is devoid of the His64 proton shuttle. A large number of aromatic/heterocyclic sulfonamides and some 5-mercapto-1,3,4-thiadiazoles were investigated as TcCA inhibitors. The aromatic sulfonamides were weak inhibitors (<i>K</i>_I values of 192 nM to 84 μM), whereas some heterocyclic compounds inhibited the enzyme with <i>K</i>_I values in the range 61.6–93.6 nM. The thiols were the most potent in vitro inhibitors (<i>K</i>_I values of 21.1–79.0 nM), and some of them also inhibited the epimastigotes growth of two <i>T. cruzi</i> strains in vivo

Effect of CA II addition to IGL-1 solution on AMPK activation and endoplasmic reticulum stress (ERS).

<p>Western blotting and densitometric analysis of pAMPK (A); GRP78 (B); p-PERK (C); p-eIF (D); IRE1alpha (E) and ATF6 (F) and immunohistochemistry of CHOP (G) in steatotic liver after 120 min of normothermic <i>ex vivo</i> perfusion. CA II supplementation (IGL-1+CAII) promoted AMPK activation and induced significant decreases in ERS parameters Without affecting CHOP expression. Ctr2: Livers flushed and perfused <i>ex-vivo</i> without cold preservation; IGL-1: livers preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h-normothermic <i>ex vivo</i> perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h-normothermic <i>ex vivo</i> perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.</p

Hepatic injury and CA II expression and activity after cold ischemia and normothermic reperfusion.

<p>(A) Hepatic injury measured as AST and ALT levels after 2h-normotermic <i>ex-vivo</i> perfusion. CA II supplementation of IGL-1 attenuated AST/ALT levels, in contrast to the IGL-1 group. (B) Hematoxylin and eosin staining. The IGL-1 group presented increased hepatic damage compared to the control group (Ctr 2), while CA II addition to IGL-1 diminished hepatic damage compared to the IGL-1 group; (C) CA II protein expression by Western blot analyses. CA II expression was increased in the IGL-1+CAII group compared to Ctr 2 and IGL-1 groups; (D) CA II activity levels in fatty livers after 2h-normothermic reperfusion. CA II activity levels decreased in livers preserved in IGL-1 and IGL-1+CAII groups when compared to the control group (Ctr 2); ATP quantitation: ATP levels was partially restored in liver preserved in IGL-1 supplemented with CA II. Ctr 2: Livers flushed and perfused <i>ex-vivo</i> without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and then subjected to 2h-normothermic <i>ex vivo</i> perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h-normothermic <i>ex vivo</i> perfusion * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.</p

Effect of CA II addition to IGL-1 solution in hepatic function.

<p>Hepatic function expressed as bile output (A) and percentage of bromosulfophthalein (BSP) excretion in bile (B) after 120 min of fatty liver normothermic <i>ex vivo</i>perfusion. CA II supplementation (IGL-1+CAII) significantly enhanced bile production and BSP clearance compared to the IGL-1 group. Ctr 2: Livers flushed and perfused <i>ex-vivo</i> without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h-normothermic <i>ex vivo</i> perfusion. IGL-1+CAII: livers preserved in IGL-1 solution (4°C, 24 h) enriched with CA II (10 ug/ml) and subjected to 2h- normothermic <i>ex vivo</i> perfusion; * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.</p

Effects of CA II addition to IGL-1 solution in liver apoptosis.

<p>Liver apoptosis, measured as cleaved caspase 9/ total caspases 9 (A), cleaved caspase 3/ total caspase 3 (B) and TUNEL assay (C) in steatotic livers after 120 min of normothermic “ex vivo” perfusion. CA II addition to IGL-1 solution reduced activation of liver caspases 3 and 9 and diminished TUNEL-positive cells. Ctr 2: Liver flushed and perfused “ex-vivo” without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h- normothermic “ex vivo” perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h-normothermic “ex vivo” perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.</p

MAPK activation changes after enrichment of IGL-1 solution with CA II.

<p>Western blotting and densitometric analysis of p-38 (A); p-ERK (B) and p-JNK (C) in steatotic liver after 120 min of normothermic “ex vivo” perfusion. CA II addition to IGL-1 solution prevented MAPK activation. Ctr 2: Liver flushed and perfused “ex-vivo” without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h-normothermic “ex vivo” perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h- normothermic “ex vivo” perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.</p

Novel Carbonic Anhydrase Inhibitors with Dual-Tail Core Sulfonamide Show Potent and Lasting Effects for Glaucoma Therapy

Glaucoma, a leading cause of irreversible vision loss worldwide, is characterized by elevated intra­ocular pressure (IOP), a well-established risk factor across all its forms. We present the design and synthesis of 39 novel carbonic anhydrase inhibitors by a dual-tailed approach, strategically crafted to interact with distinct hydrophobic and hydrophilic pockets of CA active sites. The series was investigated against the CA isoforms implicated in glaucoma (hCA II, hCA IV, and hCA XII), and the X-ray crystal structures of compounds 25a, 25f, and 26a with CA II, along with 14b in complex with a hCA XII mimic, were determined. Selected compounds (14a, 25a, and 26a) underwent evaluation for their ability to reduce IOP in rabbits with ocular hypertension. Derivative 26a showed significant potency and sustained IOP-lowering effects, surpassing the efficacy of the drugs dorzolamide and bimatoprost. This positions compound 26a as a promising candidate for the development of a novel anti-glaucoma medication