Natural product myricetin is a pan-KDM4 inhibitor which with poly lactic-co-glycolic acid formulation effectively targets castration-resistant prostate cancer.

BACKGROUND: Castration-resistant prostate cancer (CRPC) with sustained androgen receptor (AR) signaling remains a critical clinical challenge, despite androgen depletion therapy. The Jumonji C-containing histone lysine demethylase family 4 (KDM4) members, KDM4A‒KDM4C, serve as critical coactivators of AR to promote tumor growth in prostate cancer and are candidate therapeutic targets to overcome AR mutations/alterations-mediated resistance in CRPC. METHODS: In this study, using a structure-based approach, we identified a natural product, myricetin, able to block the demethylation of histone 3 lysine 9 trimethylation by KDM4 members and evaluated its effects on CRPC. A structure-based screening was employed to search for a natural product that inhibited KDM4B. Inhibition kinetics of myricetin was determined. The cytotoxic effect of myricetin on various prostate cancer cells was evaluated. The combined effect of myricetin with enzalutamide, a second-generation AR inhibitor toward C4-2B, a CRPC cell line, was assessed. To improve bioavailability, myricetin encapsulated by poly lactic-co-glycolic acid (PLGA), the US food and drug administration (FDA)-approved material as drug carriers, was synthesized and its antitumor activity alone or with enzalutamide was evaluated using in vivo C4-2B xenografts. RESULTS: Myricetin was identified as a potent α-ketoglutarate-type inhibitor that blocks the demethylation activity by KDM4s and significantly reduced the proliferation of both androgen-dependent (LNCaP) and androgen-independent CRPC (CWR22Rv1 and C4-2B). A synergistic cytotoxic effect toward C4-2B was detected for the combination of myricetin and enzalutamide. PLGA-myricetin, enzalutamide, and the combined treatment showed significantly greater antitumor activity than that of the control group in the C4-2B xenograft model. Tumor growth was significantly lower for the combination treatment than for enzalutamide or myricetin treatment alone. CONCLUSIONS: These results suggest that myricetin is a pan-KDM4 inhibitor and exhibited potent cell cytotoxicity toward CRPC cells. Importantly, the combination of PLGA-encapsulated myricetin with enzalutamide is potentially effective for CRPC.

Pholiota nameko Polysaccharides Promotes Cell Proliferation and Migration and Reduces ROS Content in H2O2-Induced L929 Cells.

Pholiota nameko, a type of edible and medicinal fungus, is currently grown extensively for food and traditional medicine in China and Japan. It possesses various biological activities, such as anti-inflammatory, anti-hyperlipidemia and antitumor activities. However, P. nameko has rarely been discussed in the field of dermatology; identifying its biological activities could be beneficial in development of a new natural ingredient used in wound care. To evaluate its in vitro wound healing activities, the present study assessed the antioxidant and anti-collagenase activities of P. nameko polysaccharides (PNPs) prepared through fractional precipitation (40%, 60% and 80% (v/v)); the assessments were conducted using reducing power, hydroxyl radical scavenging activity, dichloro-dihydro-fluorescein diacetate and collagenase activity assays. The ability of PNPs to facilitate L929 fibroblast cell proliferation and migration was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scratch assays. The findings indicated that, among all fractions, PNP-80 showed the best antioxidant and anti-collagenase activity, as measured by their reducing power (IC50 of PNP-80 was 2.43 ± 0.17 mg/mL), the hydroxyl radical scavenging (IC50 of PNP-80 was 2.74 ± 0.11 mg/mL) and collagenase activity assay, and significantly reduced cellular ROS content, compared with that of H2O2-induced L929 cells. Moreover, PNP-80 significantly promoted L929 fibroblast proliferation and migration, compared with the control group. Overall, we suggested that PNP-80 could be a promising candidate for further evaluation of its potential application on wound healing.

Characterization and Antioxidant Activities of Yellow Strain Flammulina velutipes (Jinhua Mushroom) Polysaccharides and Their Effects on ROS Content in L929 Cell.

Yellow strain Flammulina velutipes, which is known as Jinhua mushroom in Taiwan, has become popular among customers due to its distinct texture that is utterly different from white strain F. velutipes. However, there has been little study on the physicochemical properties, antioxidant activities, and biological functions of yellow strain F. velutipes polysaccharides (FVYs). The specific aims of this study are to evaluate and compare the physicochemical properties, antioxidant activities, and biological functions of FVYs and white strain F. velutipes polysaccharides (FVWs) in order to select the strain appropriate for cosmetic ingredient. The FVYs and FVWs were prepared by fractional precipitation (40%, 60%, and 80%). According to the results, FVY-80 showed the greatest antioxidant activities based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) (IC50 = 2.22 mg/mL) and 2,2' -azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical assay (IC50 = 2.04 mg/mL). None of the fractions exhibited cytotoxicity toward L929 cell under a concentration of 500 µ g/mL. FVY-80 significantly reduced the reactive oxygen species (ROS) content in L929 cell by 55.96%, as compared with the H2O2-induced L929 cell, according to the dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay. In conclusion, we suggest that FVY-80 is the best source for a cosmetics ingredient.

Dual Kit/Aur Inhibitors as Chemosensitizing Agents for the Treatment of Melanoma: Design, Synthesis, Docking Studies and Functional Investigation.

Melanoma is the most serious form of skin cancer but its medication is still far from being safe and thoroughly effective. The search of novel therapeutic approaches represents therefore a health emergency to push through eagerly. In this study, we describe a novel class of dual c-Kit/Aur inhibitors, characterized by a 1,2,4-triazole core and developed by a structure-based optimization of a previously developed hit, and report the evidence of their significance as drug candidates for the treatment of melanoma. Compound 6a, merging the best inhibitory profile against the target kinases, showed anti-proliferative efficacy against the human melanoma cell lines A2058, expressing the BRAF V600D mutation, and WM266-4, expressing BRAF V600E. Significantly, it displayed also a highly synergistic profile when tested in combination with vemurafenib, thus proving its efficacy not only per se but even in a combination therapy, which is nowadays acknowledged as the cornerstone approach of the forthcoming tumour management.

Ipomoelin, a jacalin-related lectin with a compact tetrameric association and versatile carbohydrate binding properties regulated by its N terminus.

Many proteins are induced in the plant defense response to biotic stress or mechanical wounding. One group is lectins. Ipomoelin (IPO) is one of the wound-inducible proteins of sweet potato (Ipomoea batatas cv. Tainung 57) and is a Jacalin-related lectin (JRL). In this study, we resolved the crystal structures of IPO in its apo form and in complex with carbohydrates such as methyl α-D-mannopyranoside (Me-Man), methyl α-D-glucopyranoside (Me-Glc), and methyl α-D-galactopyranoside (Me-Gal) in different space groups. The packing diagrams indicated that IPO might represent a compact tetrameric association in the JRL family. The protomer of IPO showed a canonical ß-prism fold with 12 strands of ß-sheets but with 2 additional short ß-strands at the N terminus. A truncated IPO (ΔN10IPO) by removing the 2 short ß-strands of the N terminus was used to reveal its role in a tetrameric association. Gel filtration chromatography confirmed IPO as a tetrameric form in solution. Isothermal titration calorimetry determined the binding constants (K(A)) of IPO and ΔN10IPO against various carbohydrates. IPO could bind to Me-Man, Me-Glc, and Me-Gal with similar binding constants. In contrast, ΔN10IPO showed high binding ability to Me-Man and Me-Glc but could not bind to Me-Gal. Our structural and functional analysis of IPO revealed that its compact tetrameric association and carbohydrate binding polyspecificity could be regulated by the 2 additional N-terminal ß-strands. The versatile carbohydrate binding properties of IPO might play a role in plant defense.

Crystal structure of tarocystatin-papain complex: implications for the inhibition property of group-2 phytocystatins.

Tarocystatin (CeCPI) from taro (Colocasia esculenta cv. Kaohsiung no. 1), a group-2 phytocystatin, shares a conserved N-terminal cystatin domain (NtD) with other phytocystatins but contains a C-terminal cystatin-like extension (CtE). The structure of the tarocystatin-papain complex and the domain interaction between NtD and CtE in tarocystatin have not been determined. We resolved the crystal structure of the phytocystatin-papain complex at resolution 2.03 Å. Surprisingly, the structure of the NtD-papain complex in a stoichiometry of 1:1 could be built, with no CtE observed. Only two remnant residues of CtE could be built in the structure of the CtE-papain complex. Therefore, CtE is easily digested by papain. To further characterize the interaction between NtD and CtE, three segments of tarocystatin, including the full-length (FL), NtD and CtE, were used to analyze the domain-domain interaction and the inhibition ability. The results from glutaraldehyde cross-linking and yeast two-hybrid assay indicated the existence of an intrinsic flexibility in the region linking NtD and CtE for most tarocystatin molecules. In the inhibition activity assay, the glutathione-S-transferase (GST)-fused FL showed the highest inhibition ability without residual peptidase activity, and GST-NtD and FL showed almost the same inhibition ability, which was higher than with NtD alone. On the basis of the structures, the linker flexibility and inhibition activity of tarocystatins, we propose that the overhangs from the cystatin domain may enhance the inhibition ability of the cystatin domain against papain.