Identification of a selective inhibitor of murine intestinal alkaline phosphatase (ML260) by concurrent ultra-high throughput screening against human and mouse isozymes

Abstract

Alkaline phosphatase (AP) isozymes are present in a wide range of species from bacteria to man and are capable of dephosphorylation and transphosphorylation of a wide spectrum of substrates in vitro. In humans, four AP isozymes have been identified—one tissue-nonspecific (TNAP) and three tissue-specific—named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) APs. Modulation of activity of the different AP isozymes may have therapeutic implications in distinct diseases and cellular processes. For instance, changes in the level of IAP activity can affect gut mucosa tolerance to microbial invasion due to the ability of IAP to detoxify bacterial endotoxins, alter the absorption of fatty acids and affect ectopurinergic regulation of duodenal bicarbonate secretion. To identify isozyme selective modulators of the human and mouse IAPs, we developed a series of murine duodenal IAP (Akp3-encoded dIAP isozyme), human IAP (hIAP), PLAP, and TNAP assays. High throughput screening and subsequent SAR efforts generated a potent inhibitor of dIAP, ML260, with specificity for the Akp3-, compared to the Akp5- and Akp6-encoded mouse isozymes.

2-Benzylidenebenzofuran-3(2H)-ones as a new class of alkaline phosphatase inhibitors: synthesis, SAR analysis, enzyme inhibitory kinetics and computational studies

“…3.1.3.1) are membrane-bound ectonucleotidases/hydrolase enzymes, which are ubiquitous in nature from bacteria to mammals, with the ability to perform signicant physiological functions that accelerate the hydrolysis of esters containing phosphate groups. [1][2][3][4][5][6][7][8][9] They are responsible for the effective removal of phosphate groups from alkaloids, nucleotides and proteins in alkaline medium. 10 APs are dimeric non-specic enzymes with regard to the substrate, and thus hydrolyze various substrates.…”

Isonicotinohydrazones as inhibitors of alkaline phosphatase and ecto‐5′‐nucleotidase

“…L‐phenylalanine, imidazole, histamine, and theophylline are uncompetitive inhibitors of alkaline phosphatase, while phosphate, phosphoethanolamine, and phenylphosphonate inhibit alkaline phosphatase competitively . Levamisole derivatives have also been found to be selective inhibitors of TNAP, while selective inhibitors of IAP such as ML260 ( N ‐(2,5‐dimethylphenyl)‐2‐(2‐oxo‐2,3‐dihydro‐1,3‐benzoxazole‐6‐sulfonamide acetamide) have also been reported …”

Synthesis and docking studies of N‐(5‐(alkylthio)‐1,3,4‐oxadiazol‐2‐yl)methyl)benzamide analogues as potential alkaline phosphatase inhibitors

“…GCAP is also known as placental like ALP. Hyper alkaline phosphates level causes sepsis (risk factors of cancer), whereas diffuse metastatic liver cancer (Ardecky et al, 2014;Jeesook Park & Kim, 2013). Tissue-nonspecific ALP is not expressed in any particular tissue therefore, called as the tissue-nonspecific.…”

“…If IAP gets elevated then it causes problems such as obesity caused by inappropriate deposition of fats within the cell (Buchet, Millán, & Magne, 2013), neurological disorders (Fonta, Barone, Martinez, & Négyessy, 2015;Pan et al, 2017;Sebastian-Serrano et al, 2015), chronic kidney disease (Debray et al, 2013), breast cancer, prostate cancer (Lin, Huang, Zeng, & Wu, 2019;Miao et al, 2004;Ooi, Shiraki, Morishita, & Nobori, 2007), congestion or obstruction of the biliary tract, also damage kidney, and liver. Hyper alkaline phosphates level causes sepsis (risk factors of cancer), whereas diffuse metastatic liver cancer (Ardecky et al, 2014;Jeesook Park & Kim, 2013). ALP isozymes potent inhibitors are involved to cure the bones disease; atherosclerosis, renal osteodystrophy, osteoarthritis, rickets (Millán, 2013;Yang, Oh, & Pandher, 2011), cholecystitis (inflammation of the gall bladder), and myocardial infarction.…”