Studies On The Phloroglucinol Derivatives And Antifungal Activity Of Dryopteris Crassirhizoma Nakai

Dryopteris crassirhizoma Nakai, which is the dry root and petiole residue of Dryopterideceae of Dryopteris Adanson, is harvested in autumn and dried after slashing the petioles, fibrous root and filtering sediment. It is originally recorded from Shennong’s Herbal. It tastes at first light and slightly astringent, followed by increasing bitterness. It is cold and slightly poisonous, used as styptic and as treatments for fever and intestinal worms. Dryopteris crassirhizoma Nakai, mainly distributed in North China such as Hebei Province, Inner Mongolia, Manchurian and other areas, is recorded by the 2015 edition of “People’s Republic of China Pharmacopoeia”. As a common traditional Chinese medicine herb with a variety of pharmacological activities, Dryopteris crassirhizoma Nakai has anti-viral, anti-malarial, anti-tumor, anti-cancer, deworming, styptic, anti-bacteria and other clinical applications.At present, the research on Dryopteris crassirhizoma Nakai focuses mainly on the anti-bacterial infection, anti-malaria, anti-tumor and other direction, yet rarely on the inhibition of fungi, especially common dermatophytes. Our previous study found that, phloroglucinols inhibited superficial fungal infection significantly and Dryopteris crassirhizoma Nakai was abundant in phloroglucinols. Therefore, in this study, extraction and separation of phloroglucinols activities from Dryopteris crassirhizoma Nakai were investigated. The anti-fungal activities of the purified constituents were studied.The main research contents were as follows:1. Monomeric compounds were isolated from Dryopteris crassirhizoma Nakai by the method of silica gel column chromatography, Sephadex LH-20 gel, recrystallization, ODS, HPLC and other modern separation techniques. Their structures were elucidated by their physical chemical constants, chromogenic reaction and spectroscopic analysis.2. The anti-fungal activities of 3 kinds of phloroglucinol were investigated by applying inhibition experiment on Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum gypseum. And corresponding MICs were determined through the M38-A2 Micro Dilution Method.The results were as follows:1. 12 monomeric compounds were isolated from Dryopteris crassirhizoma Nakai and structures were elucidated by their physical chemical properties, chromogenic reaction and spectroscopic analysis. These monomeric compounds were 10 phloroglucinols: Albaspidin AA(Y2), Trisflavaspidic acid ABB(Y4), Albaspidin AP(Y5), Flavaspidic acid PB(Y6), Aspidin AB(Y7), Albaspidin PP(Y8), Filixic acid ABP(Y9), Desapidin AB(Y10), Albaspidin PB(Y11), Flavaspidic acid AB(Y12); and 1 fatty acid compound,(10E,12E)-9-oxooctadeca-10,12-dienoic acid(Y1); 1 saturated aliphatic alcohol, Pentacosanol(Y3).2. The MICs of 3 monomeric compounds to Trichophyton mentagrophytes were all higher than 160μg/m L; The MICs of 3 monomeric compounds to Trichophyton rubrum were as follows: Trisflavaspidic acid ABB(160μg/m L), Albaspidin AA(60μg/m L), Flavaspidic acid AB(70μg/m L); The MICs of 3 monomeric compounds to Microsporum canis were as follows: Trisflavaspidic acid ABB(140μg/m L), Albaspidin AA(90μg/mL), Flavaspidic acid AB(80μg/m L).The conclusions were as follows:12 monomeric compounds were isolated from Dryopteris crassirhizoma Nakai, including 10 phloroglucinols, 1 fatty acid compound and 1 saturated aliphatic alcohol. Compound Y1, Y10 and Y11 were isolated from this herb firstly; all 3 monomeric compounds were effective in anti-fungi activity test. For Trichophyton rubrum, order of anti-fungal activity was: Albaspidin AA>Flavaspidic acid AB>Trisflavaspidic acid ABB and for Microsporum canis was: Flavaspidic acid AB>Albaspidin AA>Trisflavaspidic acid ABB.