title: Hyaluronic acid-nimesulide conjugates as anticancer drugs against CD44-overexpressing HT-29 colorectal cancer in vitro and in vivo.
abstract: Carrier-mediated drug delivery systems are promising therapeutics for targeted delivery and improved efficacy and safety of potent cytotoxic drugs. Nimesulide is a multifactorial cyclooxygenase 2 nonsteroidal anti-inflammatory drug with analgesic, antipyretic and potent anticancer properties; however, the low solubility of nimesulide limits its applications. Drugs conjugated with hyaluronic acid (HA) are innovative carrier-mediated drug delivery systems characterized by CD44-mediated endocytosis of HA and intracellular drug release. In this study, hydrophobic nimesulide was conjugated to HA of two different molecular weights (360 kDa as HA with high molecular weight [HAH] and 43kDa as HA with low molecular weight [HAL]) to improve its tumor-targeting ability and hydrophilicity. Our results showed that hydrogenated nimesulide (N-[4-amino-2-phenoxyphenyl]methanesulfonamide) was successfully conjugated with both HA types by carbodiimide coupling and the degree of substitution of nimesulide was 1%, which was characterized by1H nuclear magnetic resonance 400 MHz and total correlation spectroscopy. Both Alexa Fluor®647 labeled HAH and HAL could selectively accumulate in CD44-overexpressing HT-29 colorectal tumor area in vivo, as observed by in vivo imaging system. In the in vitro cytotoxic test, HA-nimesulide conjugate displayed >46% cell killing ability at a nimesulide concentration of 400 µM in HT-29 cells, whereas exiguous cytotoxic effects were observed on HCT-15 cells, indicating that HA-nimesulide causes cell death in CD44-overexpressing HT-29 cells. Regarding in vivo antitumor study, both HAL-nimesulide and HAH-nimesulide caused rapid tumor shrinkage within 3 days and successfully inhibited tumor growth, which reached 82.3% and 76.4% at day 24 through apoptotic mechanism in HT-29 xenografted mice, without noticeable morphologic differences in the liver or kidney, respectively. These results indicated that HA-nimesulide with improved selectivity through HA/CD44 receptor interactions has the potential to enhance the therapeutic efficacy and safety of nimesulide for cancer treatment.
authors.affiliation: Department of Chemistry.
authors.forename: You-Sin
authors.initials: YS
authors.lastname: Jian
authors.affiliation: Department of Chemistry.
authors.forename: Ching-Wen
authors.initials: CW
authors.lastname: Chen
authors.affiliation: PhD Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, Taichung.
authors.forename: Chih-An
authors.initials: CA
authors.lastname: Lin
authors.affiliation: Department of Chemistry.
authors.forename: Hsiu-Ping
authors.initials: HP
authors.lastname: Yu
authors.affiliation: Preclinical Development Research Department, Holy Stone Healthcare Co., Ltd., Taipei.
authors.forename: Hua-Yang
authors.initials: HY
authors.lastname: Lin
authors.affiliation: Department of Chemistry.
authors.forename: Ming-Yuan
authors.initials: MY
authors.lastname: Liao
authors.affiliation: Department of Chemistry.
authors.forename: Shu-Huan
authors.initials: SH
authors.lastname: Wu
authors.affiliation: Department of Chemistry.
authors.forename: Yan-Fu
authors.initials: YF
authors.lastname: Lin
authors.affiliation: Department of Chemistry; PhD Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, Taichung; Research Center for Sustainable Energy and Nanotechnology; Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung, Taiwan.
authors.forename: Ping-Shan
authors.initials: PS
authors.lastname: Lai
mesh_terms: D000272:Adipates / Q000737:chemistry; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000188:drug therapy* / Q000473:pathology; D053903:DNA Breaks, Double-Stranded; D016503:Drug Delivery Systems; D005260:Female; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000737:chemistry*; D020287:In Situ Nick-End Labeling; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008970:Molecular Weight; D066244:Proton Magnetic Resonance Spectroscopy; D013449:Sulfonamides / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000272:Adipates; D000970:Antineoplastic Agents; C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D013449:Sulfonamides; C029900:adipic acid; D006820:Hyaluronic Acid; D016650:Fluorescein-5-isothiocyanate; C012655:nimesulide
keywords: CD44; COX-2 inhibitor; colorectal cancer; hyaluronic acid; nimesulide