Studies with tissue cultures of the Chinese herbal plant, Tripterygiumwilfordii. Isolation of metabolites of interest in rheumatoid arthritis, immunosuppression, and male contraceptive activity

Abstract

A detailed study of metabolites produced by the plant cell culture line of Tripterygium wilfordii, a Chinese herbal plant, is presented. Eighteen compounds within the diterpene and triterpene families have been isolated and fully characterized. Of these, five are novel compounds, and their structures were determined by a combination of spectral analysis, chemical correlation and single crystal X-ray diffraction. The interest of these compounds in the treatment of rheumatoid arthritis, skin allergies, and for male contraception is noted. On rapporte une etude dCtaillCe des metabolites produits par une IignCe de culture cellulaire du Tripterygium wilfordii, un plante galenique chinoise. On a isole et caractCrisC 18 composks des familles des di-et des triterpknes. Cinq de ces produits sont de nouveaux composCs; on a determine leur structure par une combinaison d'analyses spectrales, de corrClations chimiques et de diffraction des rayons X par des cristaux uniques. On a not6 l'intCr&t de ces composCs pour le traitement de l'arthrite rhumatismale, d'allergies de la peau et comme contraceptifs pour les males.[Traduit par la rkdaction]Tripterygium wilfordii Hook f is a perennial twining vine of the family Celastraceae, which is cultivated in many parts of southern China such as Zhejiang, Anhui, Jiangxi, Fujian, and Guangdong provinces and also in Taiwan. The herb is commonly known in China as Lei Gong Teng (Thunder God vine) or Mang Cao (rank grass). Its use in Chinese traditional medicine dates back many centuries, and it is first mentioned in the Saint Peasant's Scripture of Materia Medica (1), written about two thousand years ago, as being used for the treatment of fever, chills, oedema, and carbuncle. Chinese gardeners used the powdered root to protect their crops from chewing insects. Most recently, crude extracts and refined extracts (a so-called multi-glycoside extract, or GTW) have been used increasingly to treat such disorders as rheumatoid arthritis, ankylosing spondilitis, and a variety of dermatological disorders (2-4).In 1972, Morris Kupchan et al. first isolated the novel diterpenoid triepoxides, tripdiolide ( I ) , triptolide (2), and triptonide (2a) from the roots of Triptetygium wilfordii (5) and showed that 1 and 2 have significant antileukaemic activity. The biological properties of 1 and 2 stimulated studies in our laboratory directed at developing a plant cell culture line that would hopefully produce these compounds in higher yield and under laboratory controlled conditions. The earlier studies, with these objectives in mind, have been published (6, 7) and the stable cell line, coded as TRP4a, was shown to produce 1 in yields 40 times that of the living plant.As these studies were underway, we became aware of more recent research data from various laboratories in the People's Republic of China that revealed an increasing interest in the clinical use of extracts from T. wilfordii plants. Various disorders (rheumatoid arthritis, skin disorders) have been 'Author to whom correspondence may be address...

Biosynthesis of Diterpenoids in Tripterygium Adventitious Root Cultures

“…In our hands, the production of triptolide in adventitious root cultures also has been reliable (grown continuously for more than 2 years) and readily scalable (from 50-to 500-mL volumes within weeks). Others have scaled up Tripterygium cultures to 10-to 20-L bioreactors (Kutney et al, 1992;Miao et al, 2014); therefore, it appears that typical shortcomings of tissue culture (reliability and scalability) have already been addressed.…”

“…Tissue cultures represent a promising alternative for the production of high-value plant metabolites. Early studies with T. wilfordii suspension cultures were designed to unravel the structures of small-molecule constituents (Kutney et al, 1981(Kutney et al, , 1992(Kutney et al, , 1993Kutney and Han, 1996;Nakano et al, 1997Nakano et al, , 1998. Similarly, hairy root cultures were initially employed for phytochemical investigations (Nakano et al, 1998).…”

Cytotoxic and Anti-HIV Principles from the Rhizomes of Begonia nantoensis

“…18,19) Fraction 4 was chromatographed on silica gel column with a gradient of i-Pr 2 O-MeOH (49 : 1 to pure MeOH) to give 2-(2-hydroxytricosanoylamino)-1,3,4-hexadecanetriol (28 mg), 20) methylparaben (1 mg), 21) p-hydroxybenzaldehyde (4 mg), 21) and trans-docosanylferulate (15 mg). 22) Fraction 5, on repeated chromatography on silica gel column with a gradient of i-Pr 2 O-MeOH (49 : 1 to pure MeOH) afforded cucurbitacin B (4, 60 mg), 23) dihydrocucurbitacin B (5, 5 mg), 24) cucurbitacin E (6, 18 mg), 4) dihydrocucurbitacin E (7, 4 mg), 23) cucurbitacin I (8, 3 mg), 4) dihydrocucurbitacin I (1 mg), 4) 3b,22a-dihydroxyolean-12-en-29-oic acid (10, 5 mg), 25) indole-3-carboxaldehyde (15 mg), 26) indole-3-carboxylic acid (11, 4 mg), 27) 5,7-dihydroxychromone (12, 4 mg), 28) (S)-N-(1-hydroxymethyl-2-phenylethyl)-benzamide (5 mg), 29) vanillic acid (4 mg), 9) piperonylic acid (1 mg), 30) benzoic acid (20 mg), 31) and caffeic acid (2 mg). 15) The concentrated EtOAc layer (16 g) was subjected to column chromatography over silica gel and eluted with a gradient of i-Pr 2 O-MeOH (5 : 1 to pure MeOH) to give nine fractions.…”

Epikatonic acid from Austroplenckia populnea: structure elucidation by 2D NMR spectroscopy and X‐ray crystallography

“…The structures of compounds 1 and 6 were established with one-dimensional (1D) and twodimensional (2D) NMR techniques; compounds 2, 3 and 4 were identified by comparison through thin-layer chromatography with authentic samples and literature data 1,2 and compound 5 only with literature data. 3 The NMR data Epikatonic acid structure elucidation 367 for 6 were published by Kutney et al 4 but without chemical shift attribution.…”