title: Novel substrate-based inhibitors of human glutamate carboxypeptidase II with enhanced lipophilicity.
abstract: Virtually all low molecular weight inhibitors of human glutamate carboxypeptidase II (GCPII) are highly polar compounds that have limited use in settings where more lipophilic molecules are desired. Here we report the identification and characterization of GCPII inhibitors with enhanced liphophilicity that are derived from a series of newly identified dipeptidic GCPII substrates featuring nonpolar aliphatic side chains at the C-terminus. To analyze the interactions governing the substrate recognition by GCPII, we determined crystal structures of the inactive GCPII(E424A) mutant in complex with selected dipeptides and complemented the structural data with quantum mechanics/molecular mechanics calculations. Results reveal the importance of nonpolar interactions governing GCPII affinity toward novel substrates as well as formerly unnoticed plasticity of the S1' specificity pocket. On the basis of those data, we designed, synthesized, and evaluated a series of novel GCPII inhibitors with enhanced lipophilicity, with the best candidates having low nanomolar inhibition constants and clogD > -0.3. Our findings offer new insights into the design of more lipophilic inhibitors targeting GCPII.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic , Flemingovo náměstí, Praha 6, Czech Republic.
authors.forename: Anna
authors.initials: A
authors.lastname: Plechanovová
authors.forename: Youngjoo
authors.initials: Y
authors.lastname: Byun
authors.forename: Glenda
authors.initials: G
authors.lastname: Alquicer
authors.forename: L'ubica
authors.initials: L
authors.lastname: Skultétyová
authors.forename: Petra
authors.initials: P
authors.lastname: Mlčochová
authors.forename: Adriana
authors.initials: A
authors.lastname: Němcová
authors.forename: Hyung-Joon
authors.initials: HJ
authors.lastname: Kim
authors.forename: Michal
authors.initials: M
authors.lastname: Navrátil
authors.forename: Ronnie
authors.initials: R
authors.lastname: Mease
authors.forename: Jacek
authors.initials: J
authors.lastname: Lubkowski
authors.forename: Martin
authors.initials: M
authors.lastname: Pomper
authors.forename: Jan
authors.initials: J
authors.lastname: Konvalinka
authors.forename: Lubomír
authors.initials: L
authors.lastname: Rulíšek
authors.forename: Cyril
authors.initials: C
authors.lastname: Bařinka
mesh_terms: D000954:Antigens, Surface / Q000235:genetics; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004151:Dipeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D043425:Glutamate Carboxypeptidase II / Q000037:antagonists & inhibitors* / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011789:Quantum Theory; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D000954:Antigens, Surface; D004151:Dipeptides; D008024:Ligands; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II