Composition for improving contact dermatitis containing pellitorine as an active ingredient
A technology of contact dermatitis and composition, applied in the field of contact dermatitis improvement composition, capable of solving imperfections and other problems
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Embodiment 1
[0067] Example 1. Isolation of pellitorine compound from Caulis japonica extract
[0068] After pulverizing the dried Caulis chinensis (10.2kg), extracting with 80% acetone at room temperature for 3 times and filtering, the extract was concentrated under reduced pressure to obtain 340g of extract. After the extract was concentrated under reduced pressure, 70% methanol-hexane (MeOH-Hexane), 70% methanol-chloroform (MeOH-Chloroform) was used to carry out solvent fractionation, and the fractionation was divided into 3 layers (hexane layer, chloroform layer, 70 % methanol layer). Among them, silica gel (Silica gel) column chromatography was used for the hexane layer, and the concentration of the solvent was increased according to hexane-ethyl acetate = 1:0 to 0:1. After performing TLC, it was fractionated into 17 layers. Among them, for Fr.9 The layer is subjected to DaiSogel ODS-B and medium pressure liquid chromatography (MPLC) (60% MeOH→100% MeOH, gradient system) column chr...
Embodiment 2
[0069] Example 2. Identification of the structure of pellitorine compound
[0070] Determination of the pellitorine compound obtained in Example 1 1 H and 13 C-nuclear magnetic resonance spectrum (NMR spectrum), its result is as follows:
[0071] ① 1 H-NMR (600MHz, CDCl 3 )
[0072] :δ H 7.18(1H,dd,J=10.8,15.0Hz,H-3'),6.12(1H,dd,J=10.8,15.0Hz,H-4'),6.05(1H,dt,J=6.6,15.0Hz ,H-5'),5.90(1H,br s,N-H),5.81(1H,d,J=15.0Hz,H-2'),3.15(2H,t,J=6.6Hz,H-1), 2.13(2H,m,H-6'),1.80(1H,m,H-2'),1.41(2H,m,H-7'),1.29(4H,m,H-8',9') ,0.92(6H,d,J=6.6Hz,CH 3 -3,4),0.88(3H,t,J=7.2Hz,CH 3 -10')
[0073] ② 13 C-NMR (150MHz, CDCl 3 )
[0074] :δ c 166.5(C-1'), 143.0(C-5'), 141.1(C-3'), 128.2(C-4'), 121.8(C-2'), 46.9(C-1), 32.8(C -6'),31.3(C-8'),28.5(C-7'),28.5(C-2),22.4(C-9'),20.1(C-3,4),13.9(C-10 ')
[0075] The pellitorine compound obtained in Example 1 was determined to be (E,E)-N-(2-methylpropyl)2,4-decadienamide represented by the following Chemical Formula 1 from the above resul...
Embodiment 3
[0078] Example 3. Evaluation of the effect of pellitorine compound on regulating immune cell activity
[0079] 3-1. Isolation of splenocytes
[0080] In order to perform the experiments described below, splenocytes need to be isolated from experimental animals. That is, 6-7 week-old Balb / C mice were used as experimental animals provided by Orient Bio (Seongnam, Korea) Co., Ltd. The experimental animals were kept at 22-24° C. and the humidity was 50%, and sufficient feed and water were supplied. Spleens excised from Balb / C mice were sufficiently crushed with a mesh, and connective tissue and the like were removed with a cell strainer (SPL, Korea), followed by centrifugation to obtain splenocytes. 10 ml of ACK solvent was added to the obtained splenocytes, reacted at room temperature for 5 minutes, and after red blood cells were removed, the splenocytes were separated by centrifugation twice.
[0081] 3-2. Determination of the ability of pellitorine compounds to inhibit...
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