Studies On Homoisoflavonoids And Effective Fraction Of Danggui-beimu-kushen Formula

Homoisoflavonoids represent a particular group of flavonoids with one more carbon atom than the ordinary isoflavonoids on their skeleton. More than200homoisoflavonoids have been discovered and studied. Most of them are obtained from Liliaceae and Leguminosae plants, and some are synthetic products. Caesalpinia sappan, Polygonatum cyrtonema and Polygonatum odoratum are plants which contain homoisoflavonoids. In the first part of this thesis,11homoisoflavonoids, were isolated from the EtOAc-soluble fraction of the95%ethanol extract of the above three plants and identified by various modern spectroscopic techniques. Structures of the11homoisoflavonoids were determined as (3R)-5,7-dihydroxy-8-methyl-3-(2’-hydroxy-4’-methoxy-benzyl)-chroman-4-one (1), methylophiopogonanone B (2),5,7-dihydroxy-6-methyl-8-methoxy-3-(4’-hy-droxy-benzyl)-chroman-4-one (3),5,7-dihydroxy-6,8-dimet-hyl-3-(4’-hydroxy-benzyl)-chroman-4-one (4), sappanone A (5), intricatinol (6), bonducellin (7),7-hydroxy-3-(4’-hydroxy-benzylidene)-chroman-4-one (8),3-deoxysappanchalcone (9), protosappanin A (10), brazilin (11). Compound1was identified as a new compound and the absolute configuration at C-3was determined as 3R by theoretical TD-DFT calculation of its ECD spectrum.In the second part of this thesis, the effective fraction of Danggui-beimu-kushen Formula, recorded in Golden Chamber, was studied. Firstly, for the extraction and purifition of effective fractions from each plant material, Angelica sinensis oil was extracted by SFE-CO2, and total alkaloids of Fritillaria thunbergii and Sophora flavescens were extracted by70%ethanol and further purified by solvent partition and macroporous resin, respectively. The contents of effective components were determined to be above50%in all the corresponding effective fractions. Secondly, the aerosol of the combined effective fractions was prepared on the basis of formula screening and process optimizing. Several bottles of metered dose inhaler (MDI) were then prepared and their qualities were evaluated by some indices, such as the total number of press, the weight per press, droplet distribution, the main content per press, etc. The best formula was established as6g plant materials for each of Angelica sinensis, Fritillaria thunbergii, and Sophora flavescens,3.2ml of anhydrous alcohol,0.8ml of propanediol,0.05g of tween-80, and10.5g of R134a as the propellant. All indices listed in the Chinese Pharmacopoeia (2010) were evaluated and among them the total number of press and droplet distribution met the standard while the weight per press and the main content per press were below the standard because of the low accuracy of the valve. Finally, an early-stage COPD mouse model was established by passive smoking technique and applied for the evaluation of pharmacological effect of the combined fractions. Both the intragastric and aerosol administrations showed inhibitive activities against pulmonary inflammation and mucus hypersecretion. The aerosol administration also showed inhibitive effects against several inflammatory cytokines, including TNF-a, while the intragastric administration didn’t have inhibitive effects on TNF-a. The results showed that the combined effective fractions of the Danggui-beimu-kushen Formula could be a promising TCM new drug candidate.