title: A new potential spreading factor: Streptomyces koganeiensis hyaluronidase. A comparative study with bovine testes hyaluronidase and recombinant human hyaluronidase of the HA degradation in ECM.
abstract: BACKGROUND
  Recombinant human hyaluronidase has been used in the interstitial matrix to promote the dispersion of therapeutics. The production and isolation of an extracellular hyaluronidase from Streptomyces koganeiensis (rHyal_Sk) has recently been described.
  
  
  METHODS
  The specificity of rHyal_Sk has been assessed against heparan sulfate, chondroitin sulfates and sulfated HAs. The oligomers generated by HA degradation have been investigated by MALDI-TOF MS analysis. rHyal_Sk has been compared with BTH and PH20 in vitro, against cross-linked HA (ACP) and HA-aggrecan complex, and in vivo, by means of a diffusion assay in nude mice.
  
  
  RESULTS
  Depolymerization of HA by rHyal_Sk gave tetra-, hexa- and octasaccharides in high yields. The reaction mechanism and the high HA specificity were demonstrated. The in vivo diffusion assay, supported by the in vitro tests, evidenced an initially enhanced enzymatic activity of rHyal_Sk compared to BTH and PH20.
  
  
  CONCLUSIONS
  rHyal_Sk, compared to BTH and PH20, showed higher substrate specificity and no inhibition from GAGs sulfate, together with a superior performance for HA depolymerization in ECM. As better predictive tests for the in vivo activity of hyaluronidase we developed two assays based on the degradation of ACP or of the HA-aggrecan complex.
  
  
  CONCLUSIONS
  rHyal_Sk is a new potential spreading factor for intradermal drug administration. Hyaluronidases of distinct classes, that show equivalent activities in a common turbidimetric assay, could have different potencies and dose-efficacies in vivo which influences the therapeutic effect. The new proposed in vitro tests are designed to obtain a predictive characterization of the enzyme activity in vivo.
authors.affiliation: Fidia Farmaceutici S.p.A., via Ponte della Fabbrica 3/A, 35031 Abano Terme, (PD), Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Pavan
authors.affiliation: Fidia Farmaceutici S.p.A., via Ponte della Fabbrica 3/A, 35031 Abano Terme, (PD), Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Beninatto
authors.affiliation: Fidia Farmaceutici S.p.A., via Ponte della Fabbrica 3/A, 35031 Abano Terme, (PD), Italy.
authors.forename: Devis
authors.initials: D
authors.lastname: Galesso
authors.affiliation: Fidia Farmaceutici S.p.A., via Ponte della Fabbrica 3/A, 35031 Abano Terme, (PD), Italy.
authors.forename: Susi
authors.initials: S
authors.lastname: Panfilo
authors.affiliation: Fidia Farmaceutici S.p.A., Contrada Pizzuta snc, 96017 Local Unit Noto, (SR), Italy.
authors.forename: Susanna
authors.initials: S
authors.lastname: Vaccaro
authors.affiliation: Fidia Farmaceutici S.p.A., Contrada Pizzuta snc, 96017 Local Unit Noto, (SR), Italy.
authors.forename: Luciano
authors.initials: L
authors.lastname: Messina
authors.affiliation: Fidia Farmaceutici S.p.A., via Ponte della Fabbrica 3/A, 35031 Abano Terme, (PD), Italy. Electronic address: cguarise@fidiapharma.it.
authors.forename: Cristian
authors.initials: C
authors.lastname: Guarise
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D002417:Cattle; D005260:Female; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009844:Oligosaccharides / Q000737:chemistry*; D011133:Polysaccharide-Lyases / Q000737:chemistry*; D013302:Streptomyces / Q000201:enzymology*; D013737:Testis / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D009844:Oligosaccharides; D006820:Hyaluronic Acid; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase
keywords: Diffusion assay; Glycosaminoglycan; Hyaluronidase; Sulfated HA