Improved Surface Display of Human Hyal1 and Identification of Testosterone Propionate and Chicoric Acid as New Inhibitors

Abstract

Degradation of high molecular weight hyaluronic acid (HA) in humans is mainly catalyzed by hyaluronidase Hyal1. This enzyme is involved in many pathophysiological processes and therefore appears an interesting target for drug discovery. Until now, only a few inhibitors of human Hyal1 are known due to obstacles in obtaining active enzymes for inhibitor screening. The aim of the present work was to provide a convenient enzyme activity assay and show its feasibility by the identification of new inhibitors. By autodisplay, Escherichia coli F470 can present active Hyal1 on its surface. In this study, the inducible expression of Hyal1 on the cell surface of E. coli under the control of a rhamnose-dependent promoter (Prha) was performed and optimized. Enzyme activity per single cell was increased by a factor of 100 compared to the constitutive Hyal1 surface display, as described before. An activity of 6.8 × 10−4 mU per single cell was obtained under optimal reaction conditions. By this modified activity assay, two new inhibitors of human Hyal1 were identified. Chicoric acid, a natural compound belonging to the phenylpropanoids, showed an IC50 value of 171 µM. The steroid derivative testosterone propionate showed and IC50 value of 124 ± 1.1 µM. Both values were in the same order of magnitude as the IC50 value of glycyrrhizic acid (177 µM), one of the best known inhibitors of human Hyal1 known so far. In conclusion, we established a new enzyme activity assay for human Hyal1 and identified new inhibitors with this new assay method.

Hyaluronic Acid-Modified and Doxorubicin-Loaded Gold Nanoparticles and Evaluation of Their Bioactivity

“…Recent studies have demonstrated that the surface of cancer cells over-express the CD44 receptors on their surface, to which HA strongly binds [ 25 , 26 , 27 , 28 , 29 , 30 ]. Therefore, HA can be utilized as a target ligand for tumor-targeted drug delivery [ 31 , 32 , 33 , 34 , 35 ]. The structure of HA can be modified to produce a variety of derivatives, which not only maintains its stability, but also enhances its bio-compatibility.…”

Extraction Optimization, Antioxidant, Cosmeceutical and Wound Healing Potential of Echinacea purpurea Glycerolic Extracts

“…Previous research shows that chicoric and caftaric acid have excellent antihyaluronidase activity [ 33 ]. Furthermore, chicoric acid, the main ingredient of TPA-opt, was found to inhibit human hyaluronidase 1, the enzyme that degrades high molecular weight hyaluronic acid [ 34 ]. Other caffeic acid derivatives may also add to the beneficial effect on wound healing.…”

Plant Natural Products: A Promising Source of Hyaluronidase Enzyme Inhibitors

“…In a study, Lengers and team found chicoric acid (IC 50 = 171 μM) and testosterone propionate (IC 50 = 124 ± 1.1 μM) as strong inhibitors of Hyal1 expressed over the surface of Escherichia coli F470 which was comparable to that of glycyrrhizic acid (IC 50 = 177 μM) [68].…”