Selective cholinesterase inhibition by lanostane triterpenes from fruiting bodies of Ganoderma lucidum

Two new lanostane triterpenes, named methyl ganoderate A acetonide (1) and n-butyl ganoderate H (2), were isolated from the fruiting bodies of Ganoderma lucidum together with 16 known compounds (3-18). Extensive spectroscopic and chemical studies established the structures of these compounds as methyl 7β,15α-isopropylidenedioxy-3,11,23-trioxo-5α-lanost-8-en-26-oate (1) and n-butyl 12β-acetoxy-3β-hydroxy-7,11,15,23-tetraoxo-5α-lanost-8-en-26-oate (2). Because new compounds exhibiting specific anti-acetylcholinesterase activity are being sought as possible drug candidates for the treatment of Alzheimer's and related neurodegenerative diseases, compounds 1-18 were examined for their inhibitory activities against acetylcholinesterase and butyrylcholinesterase. All of the compounds exhibited moderate acetylcholinesterase-inhibitory activity, with IC(50) values ranging from 9.40 to 31.03μM. In contrast, none of the compounds except lucidadiol (13) and lucidenic acid N (14) exhibited butyrylcholinesterase-inhibitory activity at concentrations up to 200μM. These results indicate that these lanostane triterpenes are preferential inhibitors of acetylcholinesterase and may be suitable drug candidates.     

A distinctive ribonuclease from fresh fruiting bodies of the medicinal mushroom Ganoderma lucidum

A ribonuclease with an N-terminal sequence distinct from other mushroom ribonucleases was isolated from fresh fruiting bodies of the medicinal mushroom Ganoderma lucidum. The ribonuclease was adsorbed on DEAE-cellulose and Q-Sepharose, and unadsorbed on CM-Sepharose. It possessed a molecular mass of 42 kDa as judged by gel filtration by fast protein liquid chromatography on Superdex 75 and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its molecular mass was similar to that of straw mushroom ribonuclease but much higher compared with those of other mushroom ribonucleases. The ribonuclease was unique among mushroom ribonucleases in that it exhibited the highest potency toward poly(U), followed by poly(A). Its activity toward poly(G) and poly(C) was about one-half of that toward poly(A) and one-quarter of that toward poly(U). A pH of 4.0 and a temperature of 60 degrees C were required for optimal activity of the enzyme. The optimum pH was low compared with those reported for other mushroom ribonucleases.     

Ganodermin, an antifungal protein from fruiting bodies of the medicinal mushroom Ganoderma lucidum

A 15-kDa antifungal protein, designated ganodermin, was isolated from the medical mushroom Ganoderma lucidum. The isolation procedure utilized chromatography on DEAE-cellulose, Affi-gel blue gel, CM-Sepharose and Superdex 75. Ganodermin was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-Sepharose. Ganodermin inhibited the mycelial growth of Botrytis cinerea, Fusarium oxysporum and Physalospora piricola with an IC50 value of 15.2 microM, 12.4 microM and 18.1 microM, respectively. It was devoid of hemagglutinating, deoxyribonuclease, ribonuclease and protease inhibitory activities.     

The triterpenes and steroids from the fruiting body Ganoderma duripora

Two new triterpenes (1–2) and seven steroids (3–9) were isolated from the fruiting bodies of Ganoderma duripora. Structure elucidation of these compounds was generated by a combination of spectroscopic means (HRTOFMS, ¹H and ¹³C NMR). There are significant differences between the compounds isolated from Ganoderma duripora and from the other Ganoderma species, suggesting that we need to reconsider the classification of G. duripora according to the taxonomy of chemistry.     

Purification and characterization of lectin from fruiting body of Ganoderma lucidum: lectin from Ganoderma lucidum

A novel 114 kDa hexameric lectin was purified from the fruiting bodies of the mushroom Ganoderma lucidum. Biochemical characterization revealed it to be a glycoprotein having 9.3% neutral sugar and it showed hemagglutinating activity on pronase treated human erythrocytes. The lectin was stable in the pH range of 5-9 and temperature up to 50 degrees C. The hemagglutinating activity was inhibited by glycoproteins that possessed N-as well as O-linked glycans. Chemical modification of the G. lucidum lectin revealed contribution of tryptophan and lysine to binding activity. The thermodynamics of binding of bi- and triantennary N-glycans to G. lucidum lectin was studied by spectrofluorimetry. The lectin showed very high affinity for asialo N-linked triantennary glycan and a preference for asialo glycans over sialylated glycans. The binding was accompanied with a large negative change in enthalpy as well as entropy, indicating primarily involvement of polar hydrogen, van der Waals and hydrophobic interactions in the binding.     

The anti-androgen effect of ganoderol B isolated from the fruiting body of Ganoderma lucidum

The anti-androgenic activity of the ethanol extract of the fruiting body of Ganoderma lucidum has been previously reported. Ganoderol B with 5alpha-reductase inhibitory activity and the ability to bind to androgen receptor (AR) can inhibit androgen-induced LNCaP cell growth and suppress regrowth of the ventral prostate induced by testosterone in rats. The down-regulation of AR signaling by ganoderol B provides an important mechanism for its anti-androgenic activity. In view of the fact that PSA (prostatic specific antigen, a well-accepted prognostic indicator of prostate cancer) is down-regulated, an important implication of this study is that ganoderol B intervention strategy aimed at toning down the amplitude of androgen signaling could be helpful in controlling morbidity of prostate cancer. In conclusion, our result suggests that ganoderol B might be useful in prostate cancer and benign prostatic hyperplasia (BPH) therapy through suppressing the function of androgen and its receptor.     

灵芝子实体中两个新的天然三萜类化学成分的分离、纯化和鉴定

本文采用硅胶和MCI柱层析的方法,从灵芝Ganodermalucidum子实体中分离纯化三萜类化合物。从灵芝子实体的氯仿萃取层中,分离纯化到灵芝属中的2个新天然产物,运用现代NMR技术分析确定了它们的结构,分别为methyl7β-hydroxy-3,11,15,23-tetraoxo-5α-lanost-8-en-26-oate(methylganoderateD)(Ⅰ)和methyl12β-acetoxy-3,7,11,15-tetraoxo-5α-lanost-8-en-24-oate(methyllucidenateD)(Ⅱ)。     

Structural elucidation of the polysaccharide moiety of a glycopeptide (GLPCW-II) from Ganoderma lucidum fruiting bodies

A water-soluble glycopeptide (GLPCW-II) was isolated from the fruiting bodies of Ganoderma lucidum by DEAE-Sepharose Fast-Flow and Sephacryl S-300 High Resolution Chromatography. The glycopeptide had a molecular weight of 1.2x10(4)Da (determined by HPLC), and consisted of approximately 90% carbohydrate and approximately 8% protein as determined using the phenol-sulfuric acid method and the BCA protein assay reagent kit, respectively. The polysaccharide moiety was composed mainly of D-Glc, L-Fuc, and D-Gal in the ratio of 1.00:1.09:4.09. To facilitate structure-activity studies, the structure of the GLPCW-II polysaccharide moiety was elucidated using 1H and 13C NMR spectroscopy including COSY, TOCSY, HMBC, HSQC, and ROESY, combined with GC-MS of methylated derivatives, and shown to consist of repeating units with the following structure: [Formula: see text].     

硬孔灵芝的化学成分研究

采用硅胶柱层析法进行分离纯化,从硬孔灵芝Ganoderma duropora的氯仿萃取物中分离得到甾类化合物8种。根据波谱数据,化合物1-8结构分别被鉴定为:麦角甾醇、麦角甾-7,22-二烯-3β-醇、麦角甾-7,22-二烯-3-酮、6,9-环氧麦角甾-7,22-二烯-3β-醇、过氧麦角甾醇、3,5-二羟基麦角甾-7,22-二烯-6-酮、β-谷甾醇和胡萝卜苷。     

A new class of natural glycopeptides with sugar moiety-dependent antioxidant activities derived from Ganoderma lucidum fruiting bodies

A water-soluble glycopeptide (PGY), fractionated and purified from the aqueous extract of the fruiting bodies of Ganoderma lucidum via two-step dialysis, anion exchange, and gel permeation chromatography, was constituted of two moieties of carbohydrate and peptide. Carbohydrate characterization with component analysis, methylation analysis, periodate oxidation, Smith degradation, enzymic hydrolysis, and IR and NMR experiments demonstrated that the carbohydrate moiety possessed a backbone of approximately 33 (1-->3)-linked beta-d-glucopyranosyl residues and side chains, at positions 6, of single alpha-l-arabinofuranosyl residues for every three Glcp residues in the main chain. On the basis of the results of amino acid composition and trypsin digestion, the peptide moiety, shown to consist of Arg, Ser, Ala, and Gly in a ratio of 1:1:2:2, exhibited the sequence of Ser-Arg-[(Ala)2(Gly)2] and was O-attached to the carbohydrate moiety via Ser. To contribute toward our understanding of the structure-activity relationship, a series of expected derivatives generated from PGY by trypsin digestion, debranching, and NaIO4 oxidation following reduction experiments, including PTC, DB-PGY, and PPP, were obtained. All of them, as well as PGY and a reference compound (butylated hydroxytoluene, BHT), were evaluated with two conventional antioxidant testing systems of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radical scavenging and found to have their respective antioxidant activities in a concentration-dependent manner. Comparable radical-scavenging activities observed between PTC and PGY demonstrated that the removal of Ala and Gly in a peptide moiety did not result in the variation of biological functions of PGY. However, it was very interesting to note that the scavenging activity of PPP was higher for DPPH radicals, with an SC(50) value of 116.4 +/- 5.1 microg/mL, and lower for superoxide radicals, with an SC(50) value of 205.2 +/- 14.4 microg/mL, than that of PGY with corresponding SC(50) values of 133.5 +/- 5.5 and 140.5 +/- 7.7 microg/mL, and, moreover, DB-PGY displayed the weakest scavenging potency in the tested samples, indicating that the carbohydrate moiety, in particular the side chain of nonreducing end units of Araf residues as the functional region, played a vital role in the structure and antioxidant activity. In addition, compared with the SC(50) value of BHT, PGY showed lower DPPH radical-scavenging activity but possessed higher superoxide radical-quenching potency, suggesting that it could be presented as a possible new source of natural antioxidants.     

Meroterpenoids from the fruiting bodies of higher fungus Ganoderma resinaceum

Phytochemical investigation on the fruiting bodies of Ganoderma resinaceum led to the isolation of five new meroterpenoids, namely ganoresinains A–E (1–5), and four known analogues (6–9). The new compounds were identified by extensive analyses of spectroscopic data (NMR, MS, UV, and IR) and comparison with the literature data. Compound 1 and 6 were isolated as enantiomeric mixture, which were separated over analytical chiral HPLC chromatography. Compounds 6–9 were isolated from G. resinaceum for the first time.     

Two new nortriterpenoids from the fruiting bodies of Ganoderma daqingshanense

Two new nortriterpenoids named daqingshones A (1) and B (2), along with two known ganderic acids (3 and 4), were isolated from the fruiting bodies of Ganoderma daqingshanense. Their structures were elucidated by spectroscopic data including MS, 1D and 2D NMR. Compound 2 exhibited weak anti-acetylcholinesterase (AChE) activity with IC₅₀ value of 39.2 μM.     

A new nortriterpenoid from the fruiting bodies of Ganoderma tropicum

Ganoderma tropicum has been widely used by the local folks for coronary heart disease treatment, liver protection, and sleep aid. In order to discover natural active components and tap the medical potential of G. tropicum, the chemical investigation of its fruiting bodies was carried out. This study led to the isolation of a new nortriterpenoid named 26-nor-11,23-dioxo-5α-lanost-8-en-3β,7β,15α,25-tetrol (1) and a known nortriterpenoid lucidone D (2). The structure of the new nortriterpenoid was elucidated by spectroscopic techniques including UV, IR, MS, 1D and 2D NMR spectroscopy.     

An unstructured kinetic model for the improvement of triterpenes production by Ganoderma lucidum G0119 based on nitrogen source effect

The kinetics of cell growth and triterpenes production for liquid submerged fermentation of the medicinal mushroom Ganoderma lucidum were investigated. A kinetic model was developed based on the Logistic and Luedeking-Piret equations for cell growth, substrate consumption and triterpene formation. The kinetic parameters of the model were optimized by specifically designed Runge-Kutta genetic algorithms. The mathematical model simulated the experimental data well and was capable of explaining the behavior of triterpenes production. The predictions of the kinetics from this model are very good both for normal fermentation kinetics under nitrogen limitation as well as for predictions of transitions to sluggish fermentations. The resulting model is very useful for scaling up liquid submerged fermentation of the mushroom G. lucidum and its application to the industrial production of triterpene.     

两个赤芝子实体多糖的理化特性分析及结构鉴定

赤芝子实体多糖P32A分子量为506,322,P32B分子量为287,389,两种多糖均以己糖为主构成。P32A由鼠李糖、木糖、甘露糖和葡萄糖组成,四种单糖的摩尔比为：4．3：2．6：6．3：11．4;P32B亦由鼠李糖、木糖、甘露糖和葡萄糖组成,摩尔比为：1．2：1．0：2．1：9．2。该结果显示,P32A与P32B具有较类似的糖组成,且都以葡萄糖和甘露糖为主要单糖组分。根据^13C NMR和甲基化的结果分析知P32A可能含有l→4连接的葡萄糖构成的主链,非还原末端均由葡萄糖构成,此外P32A中还有l,4-连接的甘露糖和l,3—连接的鼠李糖。P32B可能以l→3连接形成主链,部分l,3-连接葡萄糖的6位有分支,该多糖也含有由葡萄糖构成的非还原末端,与P32A类似,P32B中还含有l,4-连接的甘露糖,不同的是不含l,3—连接的鼠李糖。     

Lanostane triterpenoids from cultivated fruiting bodies of the basidiomycete Ganoderma orbiforme

A new 3,4-seco-27-norlanostane triterpene, ganoboninketal D (1), a new lanostane, (24S)-3-oxo-7α,24,25-trihydroxylanosta-8-ene (2), together with six known lanostanes (3–8), were isolated from cultivated fruiting bodies of the basidiomycete Ganoderma orbiforme. The structures were elucidated on the basis of NMR spectroscopic and mass spectrometry data, and the structures 1 and 2 were further confirmed by chemical correlations to ganoboninketal C and ganodermanondiol, respectively. All isolated compounds were inactive in the antitubercular and antimalarial activity assays against Mycobacterium tuberculosis H37Ra and Plasmodium falciparum K1, respectively, except that compound 8 exhibited weak antitubercular activity (MIC 50 μg/ml).     

灵芝子实体醇提取物的毒理研究

本文主要研究灵芝子实体醇提物的毒理学并评价其安全性,利用小鼠急性毒性试验和遗传毒性试验(Ames试验、小鼠骨髓细胞微核试验、小鼠精子畸形试验)对灵芝醇提物的毒性进行考察。试验结果显示,小鼠对灵芝醇提取物的最大耐受量是15 000mg/(kg?bw);灵芝醇提取物在有或无S9的条件下,对鼠伤寒沙门氏菌的突变型菌株TA97、TA98、TA100及TA102均无潜在致突变性;灵芝醇提取物在小鼠骨髓细胞微核试验和小鼠精子畸形试验中均呈阴性反应,不同剂量的样品组与阴性对照组之间无显著性差异。该结果表明灵芝醇提取物基本无毒性,属实际无毒物质。本研究为灵芝醇提取物的产品开发和应用提供了科学研究。     

灵芝多糖对顺铂引起的呕吐具抑制作用

以雄性昆明小鼠为实验材料,研究灵芝多糖对顺铂引起呕吐的抑制作用。实验分为5组,生理盐水+生理盐水组:腹腔注射0.9% NaCl两次,格拉司琼+顺铂组:腹腔注射格拉司琼和顺铂,生理盐水+顺铂组:腹腔注射0.9% NaCl和顺铂,灵芝多糖+生理盐水组:腹腔注射灵芝多糖和0.9% NaCl,灵芝多糖+顺铂组:腹腔注射灵芝多糖和顺铂,两种成分的注射间隔为30min。上述处理每天1次,连续5d,比较各组小鼠对高岭土的摄取量和脑内Fos蛋白表达水平。结果显示:顺铂可增加小鼠高岭土摄取量和脑内Fos蛋白表达水平,而格拉司琼和灵芝多糖可减少小鼠对高岭土的摄取量并降低脑内Fos蛋白表达水平。因此认为灵芝多糖可有效抑制顺铂引起的恶心呕吐。     

Inhibitory effect on NO production of phenolic compounds from Myristica fragrans

Three new phenolics: ((7S)-8'-(benzo[3',4']dioxol-1'-yl)-7-hydroxypropyl)benzene-2,4-diol (1), ((7S)-8'-(4'-hydroxy-3'-methoxyphenyl)-7-hydroxypropyl)benzene-2,4-diol (2) and ((8R,8'S)-7-(4-hydroxy-3-methoxyphenyl)-8'-methylbutan-8-yl)-3'-methoxybenzene-4',5'-diol (3), along with four known compounds (4-7) were isolated from the seeds of Myristica fragrans. Their chemical structures were established mainly by 1D and 2D NMR techniques and mass spectrometry. Their anti-inflammatory activity was evaluated against LPS-induced NO production in macrophage RAW264.7 cells.