Study On The Bioactive Constituents Of The Needles Of Pinus Sylvestris L.

Pinus Sylvestris (Pinus L. ) is a kind of large green arbor year occouring mainly in Dongbei Province, China. The needles of P. Sylvestris have anti– inflammatory, oxidation resisting and antivirus effects. Its major chemical constituents, including diterpenes, naphtha, flavones and so on, have been reported. Pharmacological experiments indicate that the P.E. and EtOAc extracts of this plant have significant antitumor and immunosuppressive activities. In order to search the bioactive consitituents from nature source, we began to study the chemical constituents from the needles of P. Sylvestris. By repeated column chromatography, including Silica gel, Toyopearl HW-40C and preparative HPLC, three new (1-3) and twenty-three compounds (4-26) were isolated from the EtOH extract of P. Sylvestris. By means of ¹H-NMR,¹³C-NMR and 2D-NMR spectra, the structures of new compounds were elucidated as 15-ethyl-18-methyl pinifolate (1), 15-Acetoxy-labda-8(17),13E-dien- 18-al (2), 18-Hydroxy-labd-8(17),13E-dien-15-acetate (3). Compound 4 (15- Acetoxy-labd-8(17)-en-18-oic acid) is a new natural product, and seventeen compounds were isolated from P. Sylvestris for the first time as followed: 18α-Acetoxylabd-8(17)-en-15-oic acid (8), 3β-Hydroxy-8(17),E-13-labdadien-15- oic acid (10), Dehydroabietic acid (11), 7α-Hydroxydehydro-abietic acid(12), 7β-Hydroxydehydro-abietic acid (13), 7α,18-dihydroxydehydro-abietanol (14), pimarol (16), pimaric acid (17), isopimarol (18), isopimaric acid (19), 7-oxo-13-epi-pimara-8,15-dien-18-oic acid (20),α-Cadinol (21), T-Cadinol (22), eudesm-11-en-4α-ol (23), spathulenol (24), oplopanone (25) andβ-daucosterol (26).Our previous study showed that the P.E and EtOAc extracts revealed significant antitumor activities, and the further research for isolated compounds were screened by cytotoxity effects with Hela and A549 cells. Compound 12 revealed significant inhibitory effect on Hela and A549, compounds 7 and 13 showed positively inhibitory effect on Hela, and compounds 14 and 15 revealed positively inhibitory effect on A549.In the lymphocyte transformation experimention, the P.E. and EtOAc extracts revealed the significant inhibitory effect to the concanavalin (Con A) control group (p<0.01, n=6) and positively showed immunosuppressive activity. It is necessary to take a further research on the immunosuppressive constituents from P. Sylvestris.