Structure-based design of hyaluronidase inhibitors

摘要: Hyaluronan and hyaluronidases have been used in several medical fields for many years some play a role in, e.g., meningitis, septicaemia, arthroses cancer. To further investigate the function of hyaluronan (patho)physiological processes, selective potent hyaluronidase inhibitors are required. Originated by lack such compounds, main goal this thesis was prediction lead-like structures de novo ligand design. For design with programme LUDI, homology model S. agalactiae strain 4755 (hylB4755) constructed starting from two bacterial hyaluronidases. Screening LeadQuest®, Accelrys adapted ChemACX (ChemACXF) databases resulted 1275 hits. 13 out 19 selected compounds were active on hylB4755 milli- submillimolar range. 1,3-Diacetylbenzimidazole-2-thione identified to be one most (IC50 values 5 µM 160 at physiological pH optimum pH, respectively). To validate usage aforementioned virtual screening, distribution six (physico)chemical properties (molecular weight, log P, numbers H-bond donors acceptors, rotatable bonds rings) within these analysed respect drug-likeness. The analysis revealed that LeadQuest® Vol. 1&2 1-3 suitable compound selections screening drug-like molecules. Furthermore, raw pre-filtering database elimination reactive entities outside certain molecular weight range significantly different property distributions, though covering essential pharmacological space. For bovine testicular (BTH) enzyme based crystal bee venom using MODELLER. Filtering Lead-Quest® ChemACXF ca. 5500 testing inhibition; none inhibited BTH. Additionally, ligand-based approach performed. superposition sites BTH model, chitinases A B complex very good overlap amino acids involved catalysis co-crystallised ligands. By considering substructures mimicking proposed intermediate hyaluronic acid hydrolysis introducing substituents suggested interact site enzyme, as potential BTH. Due weak inhibition pneumoniae vitamin C, more hydrophobic derivative L-ascorbic acid-6-hexadecanoate investigated proved inhibitor hylB4755, 4 µM, 100 56 respectively). binding mode determined X-ray analysis, supporting hypothesis additional interactions contribute higher affinity. predicted flexible docking FlexX suggesting alternative modes. That seems clearly favoured where long alkyl chain favourably interacts an extended, strongly channel. The hyaluronate lyase-inhibitor complexes indole moiety sulfamic 1-decyl-2-(4-sulfamoyloxy-phenyl)-1H-indol-6-yl ester C portion bind exactly same region catalytic site. aliphatic both dunk surface crevice. Using programmes LUDI GRID, regions donor, acceptor moieties may transferred into 3D pharmacophore model. known SAR 2-phenylindole derivatives observed led suggestions about benzoxazole-2-thione derivatives. Based structure lyase substrate-based hexasaccharide, novel benzoxazole-2-thiones 3-substituted N-propanoyl groups putative inhibitors. This strategy confirmed activity 3-phenylpropanoyl which potently inhibits IC50 value 15 µM.