Novel application of diarylheptanoids compound
A technology of diaryl heptane and compounds, which is applied in the field of diaryl heptane compounds, can solve the problems of no specific treatment and achieve the effects of treating sepsis, treating inflammation, and inhibiting the production of nitric oxide
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Embodiment 1 2
[0030] Example 1 Extraction, separation and identification of diaryl heptane compounds
[0031] 1.1 Instruments and materials
[0032] Perkin-Elmer FT-IR infrared spectrophotometer, KBr pellets; nuclear magnetic resonance experiments using BrukerAVANCE 400 nuclear magnetic resonance; using ES-TOFMS and ES-MS mass spectrometry using Bruker microTOF and aFinnigan LC-Q II mass spectrometer; JASCO-1020 polarimeter; high performance liquid chromatography: Agilent 1260HPLC, the preparative column uses Kromasil 100-10-C18; Merck silica gel 60 (fine than 0.063 mm) and cross-linked dextran LH-20 from Pharmacia.
[0033] The rhizomes of Curcuma comosa are in Nakhon Pathom Sakon Nakhon and Prachin Buri, Thailand, and voucher specimens (Apichart Suksamrarn, Nos. 052 and 074) are deposited at the Faculty of Science, Ramkhamhaeng University.
[0034] 1.2 Separation and extraction of compounds
[0035] The rhizomes of C.comosa were sliced, air-dried, ground and sequentially impregnated wit...
Embodiment 2
[0115] Example 2 The activity of diarylheptane compounds in inhibiting NO production.
[0116] 1 material
[0117] Compounds (1-37, 39-49); mouse RAW264.7 macrophages; endotoxin (LPS, 0.5 μg / mL); BAY (10 μM, positive control) NO detection kit (Beyotime, Haimen, China).
[0118] 2 main instruments
[0119] Fluorescent microplate reader (Thermo Scientific, Waltham, MA, USA)
[0120] 3 methods
[0121] 3.1 Determination of NO production by LPS-stimulated RAW264.7 macrophages
[0122] RAW264.7 macrophages were seeded in 96-well plates, and the seeding volume per well was 1×10 4 After the cells were inoculated and adhered to the wall, the compounds to be tested (1-37, 39-49) were added to incubate for 2 hours, and then LPS (final concentration: 0.5 μg / mL) was added to continue to incubate for 24 hours. Finally, take the 96-well plate culture medium and use the NO kit to detect the NO content by measuring its OD 550 To represent. Three replicate wells were taken for each expe...
Embodiment 3 2
[0132] Example 3 Inhibitory effect of diarylheptane compounds on macrophage proliferation
[0133] 1 material
[0134] Compound (1-37,39-49); mouse RAW264.7 macrophages; Alamar-Blue reagent.
[0135] 2 main instruments
[0136] Fluorescent microplate reader (Thermo Scientific, Waltham, MA, USA)
[0137] 3 methods
[0138] 3.1 Effects of compounds (1-37,39-49) on the proliferation of RAW264.7 macrophages
[0139] Inoculate RAW264.7 macrophages in 96-well plate, inoculate 1×10 per well 4 After the cells are inoculated and attached to the wall, the compounds to be tested (1-37, 39-49) are added and incubated for 24 hours. Then add Am-Blue cell proliferation and activity detection reagent (SunBio TM ) 10 μL and incubate in the incubator for 2-6 hours. When the color of the culture medium changed from indigo blue to pink, the relative fluorescence unit (RFU value) of each well was measured with a fluorescent microplate reader, the excitation light wavelength was 560nm, and t...
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