title: Nitric oxide-donating genistein prodrug: design, synthesis, and bioactivity on MC3T3-E1 cells.
abstract: To find a more potent alternative with less estrogen-related side effects for hormone replacement therapy, we designed and synthesized a nitric oxide (NO)-releasing prodrug of genistein, named NO-donating genistein (NO-G). The characteristics of NO-G were determined by melting point, NMR spectroscopy, and mass spectrometric analysis. HPLC has been used to test the new prodrug's stability. The releasing capacity of NO-G was tested by Griess reagent in vitro. The bioactivities of NO-G on proliferation, differentiation, and mineralization of the osteoblastic cell line MC3T3-E1 were determined by MTT assay, flow cytometric analysis, measurement of the alkaline phosphatase (ALP) activity and the secreted osteocalcin (OCN), and Alizarin Red-S staining. The product showed 1H NMR spectra and relative molecular mass in agreement with the designed structure, and it was stable in buffer solution. NO-G continually released low level NO within 5 h in MC3T3-E1 cells. NO-G caused a significant elevation of cell growth, ALP activity, and OCN secretion in both dose- and time-dependent manner. Furthermore, the Alizarin Red-S staining showed that NO-G promoted mineralization of MC3T3-E1 cells. These effects were all significantly greater than those of its parent drugs. The results suggested that NO-G might be a novel drug for the treatment of postmenopausal osteoporosis.
authors.affiliation: Department of Pharmacology, Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, and Department of Cardiology, Tangdu Hospital, Shaanxi, China.
authors.forename: Jiepin
authors.initials: J
authors.lastname: Wang
authors.forename: Fujun
authors.initials: F
authors.lastname: Shang
authors.forename: Ru
authors.initials: R
authors.lastname: Jiang
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Siwang
authors.initials: S
authors.lastname: Wang
authors.forename: Jin
authors.initials: J
authors.lastname: Hou
authors.forename: Menglei
authors.initials: M
authors.lastname: Huan
authors.forename: Qibing
authors.initials: Q
authors.lastname: Mei
mesh_terms: D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002087:Butyrates / Q000737:chemistry; D049109:Cell Proliferation / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004355:Drug Stability; D005434:Flow Cytometry; D019833:Genistein / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D051379:Mice; D015394:Molecular Structure; D009566:Nitrates / Q000737:chemistry*; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000737:chemistry / Q000494:pharmacology*; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D048789:Phytoestrogens / Q000737:chemistry / Q000494:pharmacology; D011355:Prodrugs / Q000737:chemistry / Q000494:pharmacology*; D016196:S Phase / Q000187:drug effects; D013997:Time Factors
chemical_list: D002087:Butyrates; D009566:Nitrates; D020030:Nitric Oxide Donors; D048789:Phytoestrogens; D011355:Prodrugs; C523358:genistein 7,4'-(bromo)butyrate; C523359:genistein 7,4'-(nitrooxy)butyrate; D015675:Osteocalcin; D009569:Nitric Oxide; D019833:Genistein; D000469:Alkaline Phosphatase