Prostate Metathesis

detection, 254 nm; flow rate, 1.0 ml/min; column temperature, 40 °C; mobile phase, linear gradient of 20–100% acetonitrile aqueous solution per 25 min followed by 100% acetonitrile for 20 min.Isolated metabolites from HPLC effluents were subjected to mass spectrometric analysis using a Finnegan Mat TSQ-70 with atmospheric pressure chemical ionization, positive mode.After staining nuclei with DAPI, cells were visualized using a fluorescence microscope (IX70; Olympus, Tokyo, Japan). After the incubation, total RNA was isolated from PZ-HPV-7 using SV Total RNA Isolation System (Promega) according to the manufacturer's instructions.

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The organic phase was recovered and dried down under reduced pressure.

The resultant residue was dissolved in acetonitrile and applied to HPLC under the following conditions: column, YMC-Pack ODS-AM (5 μm; 4.6×300 mm; YMC Co., Kyoto, Japan); u.v.

Optical absorbance at 570 nm was measured using a microplate reader (Varioskan; Thermo Scientific, MA, Waltham, USA).

The effect of 25(OH)D were produced in PZ-HPV-7 cells.

All cells were grown in a 37 °C humidified incubator with an atmosphere of 5% CO for 24 h.

After the incubation, metabolites were extracted using chloroform/methanol (3:1, v/v) from a mixture of cells and medium.

is not suitable as a therapeutic agent for cancer treatment.

Accordingly, the analogs that are less calcemic but exhibit potent anti-proliferative activity have potential as therapeutic agents.

Unfortunately this route has proved difficult to use for the programmed synthesis because of the lack of chemoselectivity at a decisive strategic level.

A second approach was developed starting from commercial 2-methyl-cyclopentane-1,3-dione, providing the required racemic precursor of the metathesis reaction in 10 steps and 8% yield.