Introduction
!Matrix metalloproteinases (MMPs) are a family of zinc-and calcium-dependent endopeptidases, which are grouped into the metzincin clan of metallopeptidases (MPs) together with other families such as ADAMs/adamalysins, astacins, fragilysins, and serralysins. MMPs are found throughout the animal and plant kingdoms, where their distribution is consistent with a Darwinian tree-based pathway [1]. Furthermore, polyplication has led to the presence of several paralogous MMP genes in the same organism: 24 in humans, 26 in sea urchins, 26 in zebrafish, 7 in sea squirts, and 2 in fruit flies. Overexpression of MMPs may cause various inflammatory, malignant, and degenerative symptoms [2]. Among the MMPs studied, gelatinase-A (MMP-2), gelatinase-B (MMP-9), and collagenase-1 (MMP-1) were reported to be responsible for the signal transduction of dermal photoaging [3], and inhibition of the activities of these two enzymes could potentially slow down skin aging. Thus, the search for bioactive compounds that can regulate the activities of MMP-1, -2, and -9 from natural resources is one of the key steps in delaying skin aging. [16]. In a preliminary biological evaluation, crude extracts of R. rosea roots exhibited inhibitory activities toward MMP-2 and collagenase at a concentration of 100 µg/mL [17]. An investigation of the active principles of this plant was thus undertaken by using a bioassayguided method. Of the tested water, n-butanol, and ethyl acetate layers, the ethyl acetate layer Abstract ! Based on the significant inhibitory activity toward matrix metalloproteinase-2 and collagenase noticed in preliminary studies, crude extracts of Rhodiola rosea were partitioned and chromatographed sequentially to afford three new compounds, 1,2,3,6-tetra-O-galloyl-4-O-p-hydroxybenzoyl-β-D-glucopyranoside (1), (E)-creoside I (2), and (R,Z)-2-methylhept-2-ene-1,6-diol (3), along with twenty-four known compounds (4-27). Their structures were determined by spectroscopic data analyses. All isolated compounds were subjected to bioactivity assays. In these, 1 specifically inhibited matrix metalloproteinase-2 activity with an IC 50 value of 16.3 ± 1.6 µM, while its analogue 1,2,3,6-tetra-O-galloyl-β-D-glucopyranonoside (17) inhibited matrix metalloproteinase-2 with an IC 50 value of 23.0 ± 4.8 µM. In the collagenase activity assay, the inhibitory effects of 1 and 17 at concentrations of both 20 and 40 µM were more potent than those of the positive control, 1,10-phenanthroline. In order to realize whether 17 could penetrate from the epidermal layer into the basal and dermal layers of the human skin to inhibit the activity of matrix metalloproteinase-2 and collagenase or not, a transdermal penetration test in nude and white mice skins was performed. Penetration percentages of 17 quantified by LC-MS were 27.8 % and 74.8 % in 24 hours, respectively.
Supporting information available online at