Structural Identification of a New Tri-p-coumaroylspermidine with Serotonin Transporter Inhibition from Safflower

Abstract

Safflower (Carthamus tinctorius L.), a member of the family Compositae, geologically distributes in all regions in China, and is utilized for producing herbal medicines for the treatment of inflammatory diseases, hyperlipemia, arteriosclerosis, gynecological disorders, and osteoporosis. 1) To date, it has been known to contain flavonoids, 2) cartharmin, safflower yellow A, quinochalone, safflomin, cartorimine, alkaloids, 3) lignans, fatty acids, 4) triterpene alcohols, 5) and serotonin derivatives.6-8) Additionally, several coumaroylspermidine analogs also were found in safflower, i.e., a principal analog N 1 ,N 5 ,N 10 -(E)-tri-p-coumaroylspermidine (EEE) and its three cis-trans isomers [N 1 ,N 5 9) indicating that that some other geometric isomers of tri-p-coumaroylspermidine could exist in the natural plant. In our previous preliminary screening, we discovered that safflower extracts (especially ethyl acetate extract (HE) and n-butyl alcohol fraction) had regulatory actions on the monoamine transports in transporter-transgenic Chinese hamster ovary (CHO) cells, that is, activations of dopamine (DA)/norepinephrine (NE) uptake and an inhibition of serotonin (5HT) uptake, 10) implying that there may be some constituent(s) acting as serotonin transporter (SERT) blocker(s). In this study, we thereby chose to isolate and characterize the structurally unknown, SERT-targeting compound(s) from the HE fraction by using an activity-directed extraction method and a series of NMR techniques. Finally, one of the active isolates was identified and elucidated as N 1 ,N 5 -(Z)-N 10 -(E)-tri-p-coumaroylspermidine (CX), a new tri-p-coumaroylspermidine as postulated previously by us. Results and DiscussionThe ethyl acetate extract of safflower (Carthamus tinctorius L.) was separated by column chromatography on silica gel and Sephadex LH-20 in combination with a concomitant bioactivity-tracing technique (detailed paradigm described in Supplemental) to afford a bioactive compound CX (designated as carthamide X by us) (Fig. 1). HPLC showed that the purity of CX sample was 95.6% (while the content of CX in HE was only 0.028%). Due to the sensitivity, validity, and feasibility, 10) our screening platform previously established by using isotope-ligand-detection methods is suggested suitable for tracing some structurally unknown/unstable compounds with a sparse content in natural plants when combined with phytochemical techniques, which could, however, be hard to be isolated by traditional phytochemical method alone.The We previously reported that safflower (Carthamus tinctorius L.) ethyl acetate extract (HE) possessed an inhibitory action on serotonin (5HT) uptake in Chinese hamster ovary (CHO) cells expressing 5HT transporter (SERT) (S6 cells). Here, HE was adopted to go through an activity-guided isolation, and then an ingredient with potent SERT inhibitory action was obtained, which was elucidated as N

Antifungal Polyamides of Hydroxycinnamic Acids from Sunflower Bee Pollen

“…29 Several methylene proton signals at δ 1.55− 1.95 (internal), δ 3.09−3.38 (bound to end amine nitrogen), and δ 3.18−3.58 (bound to internal amine nitrogen) and the corresponding carbon signals at δ 25.5−29.8, 37.8−38.3, and δ 39.7−47.7 confirmed the presence of spermidine molecules conjugated with phenolic acids. 30 Thus, the main compound found in the methanol/ethanol soluble fractions was tri-pcoumaroylspermidine 31 accompanied by other HCAA of spermidine and putrescine. LC−MS analyses confirmed exact masses of 584.27540 [M + H] + and m/z 438.23864 [M + H] + corresponding to tri-and di-p-coumaroylspermidines.…”

Natural Deep Eutectic Solvents as a New Extraction Media for Phenolic Metabolites in Carthamus tinctorius L.

“…The identification of the compounds (Figure 1) corresponding to each peak was confirmed by comparing NMR, HRESI-MS data obtained by UPLD-ESI-TOF-MS, previous papers, and 2D NMR spectra. 32,33,37 Compounds corresponding to four peaks were isolated and identified as N 1 ,N 10 ,N 5 -(Z)-tri-p-coumaroylspermidine (4), N 1 -(E)-N 5 ,N 10 -(Z)-tri-p-coumaroylspermidine (5), N 1 ,N 10 -(E)-N 5 -(Z)-tri-p-coumaroylspermidine (6), and N 1 ,N 10 ,N 5 -(E)-tri-p-coumaroylspermidine (8) (Figure 2). The compound 7 (a mixture of two compounds) and a small peak between the compounds 4 and 5 in HPLC chromatogram all have the same UV spectrum with a UV max at around 300 nm and the same molecular weight as the above-described tri-pcoumaroylspermidines, probably corresponding thus to other stereoisomers of tri-p-coumaroylspermidine.…”

Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast

“…For example, spermidine hydroxycinnamate conjugates have antioxidant [31, 32], antibacterial [33], antiviral [34], insecticidal [35] and antiparasitic [36] activities. Other studies have demonstrated the inhibitory activity of polysubstituted p -coumaroyl polyamines on brain transporters for which a genetic polymorphism is associated with psychological disorders [37, 38]. For this experiment, we selected the BAHDs p -coumaroyl-CoA:spermidine (AtSCT) and sinapoyl-CoA:spermidine (AtSDT) transferases from Arabidopsis [39], and p -coumaroyl-CoA:agmatine transferase (HvACT) from barley ( Hordeum vulgare ) [40].…”