title: Angiotensin converting enzyme versus angiotensin converting enzyme-2 selectivity of MLN-4760 and DX600 in human and murine bone marrow-derived cells.
abstract: Angiotensin-converting enzymes, ACE and ACE2, are key members of renin angiotensin system. Activation of ACE2/Ang-(1-7) pathway enhances cardiovascular protective functions of bone marrow-derived stem/progenitor cells. The current study evaluated the selectivity of ACE2 inhibitors, MLN-4760 and DX-600, and ACE and ACE2 activities in human (hu) and murine (mu) bone marrow cells. Assays were carried out in hu and mu mononuclear cells (MNCs) and huCD34(+) cells or mu-lineage-depleted (muLin(-)) cells, human-recombinant (rh) enzymes, and mu-heart with enzyme-specific substrates. ACE or ACE2 inhibition by racemic MLN-4760, its isomers MLN-4760-A and MLN-4760-B, DX600 and captopril were characterized. MLN-4760-B is relatively less efficacious and less-selective than the racemate or MLN-4760-A at hu-rhACE2, and all three of them inhibited 43% rhACE. In huMNCs, MLN-4760-B detected 63% ACE2 with 28-fold selectivity over ACE. In huCD34(+) cells, MLN-4760-B detected 38% of ACE2 activity with 63-fold selectivity. In mu-heart and muMNCs, isomer B was 100- and 228-fold selective for ACE2, respectively. In muLin(-) cells, MLN-4760-B detected 25% ACE2 activity with a pIC50 of 6.3. The racemic mixture and MLN-4760-A showed lower efficacy and poor selectivity for ACE2 in MNCs and mu-heart. ACE activity detected by captopril was 32% and 19%, respectively, in huCD34(+) and muLin(-) cells. DX600 was less efficacious, and more selective for ACE2 compared to MLN-4760-B in all samples tested. These results suggest that MLN-4760-B is a better antagonist of ACE2 than DX600 at 10 µm concentration in human and murine bone marrow cells, and that these cells express more functional ACE2 than ACE.
authors: Department of Pharmaceutical Sciences, North Dakota State University, Fargo, ND, USA.
authors: Shrinidh
authors: S
authors: Joshi
authors: Core Synthesis Facility, North Dakota State University, Fargo, ND, USA.
authors: Narayanaganesh
authors: N
authors: Balasubramanian
authors: Department of Pharmaceutical Sciences, North Dakota State University, Fargo, ND, USA; Core Synthesis Facility, North Dakota State University, Fargo, ND, USA.
authors: Goutham
authors: G
authors: Vasam
authors: Department of Pharmaceutical Sciences, North Dakota State University, Fargo, ND, USA. Electronic address: Yagna.Jarajapu@ndsu.edu.
authors: Yagna Pr
authors: YP
authors: Jarajapu
mesh_terms: D000368:Aged; D000595:Amino Acid Sequence; D000085962:Angiotensin-Converting Enzyme 2; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000378:metabolism*; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D006801:Humans; D007093:Imidazoles / Q000378:metabolism*; D007930:Leucine / Q000031:analogs & derivatives* / Q000378:metabolism; D008297:Male; D051379:Mice; D008875:Middle Aged; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: C486469:2-(1-carboxy-2-(3-(3,5-dichlorobenzyl)-3H-imidazol-4-yl)ethylamino)-4-methylpentanoic acid; D000806:Angiotensin-Converting Enzyme Inhibitors; C556151:DX600 peptide; D007093:Imidazoles; D010455:Peptides; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; C000705328:Ace2 protein, mouse; D000085962:Angiotensin-Converting Enzyme 2; D007930:Leucine
keywords: ACE; ACE2; CD34(+) cells; DX600; Lineage-depleted cells; MLN-4760