Advances on the microbial synthesis of plant-derived diterpenoids / 生物工程学报

Natural plant-derived diterpenoids are a class of compounds with diverse structures and functions. These compounds are widely used in pharmaceuticals, cosmetics and food additives industries because of their pharmacological properties such as anticancer, anti-inflammatory and antibacterial activities. In recent years, with the gradual discovery of functional genes in the biosynthetic pathway of plant-derived diterpenoids and the development of synthetic biotechnology, great efforts have been made to construct a variety of diterpenoid microbial cell factories through metabolic engineering and synthetic biology, resulting in gram-level production of many compounds. This article summarizes the construction of plant-derived diterpenoid microbial cell factories through synthetic biotechnology, followed by introducing the metabolic engineering strategies applied to improve plant-derived diterpenoids production, with the aim to provide a reference for the construction of high-yield plant-derived diterpenoid microbial cell factories and the industrial production of diterpenoids.

Chemical constituents of diterpenoids from Boswellia carterii / 中国中药杂志

This paper explored the chemical constituents of Boswellia carterii by column chromatography on silica gel, Sephadex LH-20, ODS column chromatography, and semi-preparative HPLC. The structures of the compounds were identified by physicochemical properties and spectroscopic data such as infrared radiation(IR), ultra violet(UV), mass spectrometry(MS), and nuclear magnetic resonance(NMR). Seven diterpenoids were isolated and purified from n-hexane of B. carterii. The isolates were identified as(1S,3E,7E,11R,12R)-11-hydroxy-1-isopropyl-4,8,12-trimethyl-15-oxabicyclo[10.2.1]pentadeca-3,7-dien-5-one(1),(1R,3S,4R,7E,11E)-4,8,12,15,15-pentamethyl-14-oxabicyclo[11.2.1]hexadeca-7,11-dien-4-ol(2), incensole(3),(-)-(R)-nephthenol(4), euphraticanoid F(5), dilospirane B(6), and dictyotin C(7). Among them, compounds 1 and 2 were new and their absolute configurations were determined by comparison of the calculated and experimental electronic circular dichroisms(ECDs). Compounds 6 and 7 were obtained from B. carterii for the first time.

Diterpenoid constituents in Pseudolarix amabilis and their antitumor activities in vitro / 中国中药杂志

By various chromatographic techniques and extensive spectroscopic methods, 17 abietane diterpenoids were isolated from the dichloromethane fraction of the 95% ethanol cold-soak extracts of the seeds of Pseudolarix amabilis, namely pseudoamaol A(1), 12α-hydroxyabietic acid(2), 12-methoxy-7,13-abietadien-18-oic acid(3), 13-hydroxy-8,11,13-podocarpatrien-18-oic acid(4), 15-hydroxy-7,13-abietadien-12-on-18-oic acid(5), 8(14)-podocarpen-13-on-18-oic acid(6), holophyllin K(7), metaglyptin B(8), 7α-hydroxydehydroabietinsaure-methylester(9), 7-oxodehydroabietic acid(10), 15-hydroxy-7-oxodehydroabietinsaure-methy-lester(11), 15-methoxydidehydroabietic acid(12), 7-oxo-15-hydroxy-dehydroabietic acid(13), 15-hydroxydehydroabietic acid(14), 8,11,13-abietatriene-15,18-diol(15), 8,11,13-abietatriene-15-hydroxy-18-succinic acid(16), and 7β-hydroxydehydroabie-tic acid(17). Compound 1 was a new compound. The isolated compounds were evaluated for their antitumor activities(HepG2, SH-SY5Y, K562), and compounds 8 and 17 showed potential cytotoxic activity against K562 cells, with IC_(50) values of 26.77 and 37.35 μmol·L~(-1), respectively.

Antimalarial and neuroprotective ent-abietane diterpenoids from the aerial parts of Phlogacanthus curviflorus / 中国天然药物

Six new ent-abietane diterpenoids, abientaphlogatones A-F (1-6), along with two undescribed ent-abietane diterpenoid glucosides, abientaphlogasides A-B (7-8) and four known analogs were isolated from the aerial parts ofPhlogacanthus curviflorus (P. curviflorus). The structures of these compounds were determined using high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), one-dimensional and two-dimensional nuclear magnetic resonance (NMR) spectroscopy, electronic circular dichroism (ECD) spectra, and quantum chemical calculations. Notably, compounds 5 and 6 represented the first reported instances of ent-norabietane diterpenoids from the genus Phlogacanthus. In the β-hematin formation inhibition assay, compounds 2, 4, 7-10, and 12 displayed antimalarial activity, with IC50 values of 12.97-65.01 μmol·L-1. Furthermore, compounds 4, 5, 8, and 10 demonstrated neuroprotective activity in PC12 cell injury models induced by H2O2 and MPP+.

Abietane diterpenoids and iridoids from Caryopteris mongolica / 中国天然药物

Six new abietane diterpenoids (1-6) and five undescribed iridoids (7-11) have been isolated from the aerial parts of Caryopteris mongolica. The intricate structural characterization of these compounds was meticulously undertaken using an array of advanced spectroscopic techniques. This process was further enhanced by the application of DP4+ probability analyses and electronic circular dichroism (ECD) calculations. Following isolation and structural elucidation, the cytotoxicity of these compounds was evaluated. Among them, compound 3 stood out, displaying significant cytotoxic activity against HeLa cells with an IC50 value of 7.83 ± 1.28 μmol·L-1. Additionally, compounds 1, 2, 4, 9, and 10 manifested moderate cytotoxic effects on specific cell lines, with IC50 values ranging from 11.7 to 20.9 μmol·L-1.

Diterpenoids of Tripterygium wilfordii decrease inflammatory response of macrophage by inhibiting multiple signaling pathways / 中国药理学通报

Aim To study the anti-inflammatory activ¬ity of diterpenes from Tripterygium wilfordii on lipopo- lysaccharide ( LPS)-induced macrophage and its mech¬anism. Methods MTT assay was used to detect the cytotoxicity of compounds. The Griess method was used to detect the NO on LPS-induced RAW264. 7 cells. ELISA was applied to determine the contents of inter- leukin 6 (IL-6) , tumor necrosis factor a ( TNF-a ) , interleukin lp (IL-lfj) and interleukin 18 (IL-18) in cell culture supernatant. Western blot was used to de¬tect IkBcx, .INK, ERK, p38, STAT3 and their phos-phorylation in LPS-induced RAW264.7, as well as the effect on COX-2, iNOS, NLRP3, caspase-1 , cleaved- caspase-1. Flow cytometry was employed to detect the effects of compounds on the phagocytosis of RAW 264. 7 cells. Results Hypoglicin II (1) and ent-pimara-8 (14) , 15-diene-19-ol (6) , two diterpenoid compounds from Tripterygium wilfordii could effectively inhibit the expression of inflammatory mediators ( COX-2 and iN- OS) and inflammatory cytokines (IL-6, IL-lp, IL- 18) in LPS-induced RAW264. 7 cells. Further re¬search found that the phosphorylation of IkBcx , JNK, ERK, P38, STAT3 and NLRP3 was all inhibited; however, there was no significant effect on the expres¬sion of IkBcx, JNK, ERK, P38 and STAT3. At the same time, they also inhibited the phagocytosis of mac-rophages. Conclusions The anti-inflammatory mecha¬nism of Tripterygium wilfordii diterpenoids 1 and 6 might be through inhibiting the production of NLRP3 inflammatory bodies, inflammatory mediators (COX-2 and iNOS) and inflammatory cytokines (IL-6, IL-lp and IL-18) , which is closely related to inhibiting the activation of MAPK, NF-kB and STAT3 pathway.

Research progress in tigliane-type macrocyclic diterpenoids / 中国中药杂志

Tigliane type macrocyclic diterpenoids with special structures and diverse bioactivities are mainly extracted from plants of Euphorbiaceae and Thymelaeaceae. According to the different functional groups, they can be classified into types of phorbol esters, C-4 deoxyphorbol esters, C-12 deoxyphorbol esters, C-16 or C-17 substituted phorbol esters and others. Most of them present promising antiviral activities and cytotoxic activities and are expected to be developed as candidates for anti-AIDS, anti-tuberculosis, and anti-tumor clinical trials, demonstrating great potential for the application in healthcare. This paper reviews 115 novel tigliane-type diterpenoids discovered since 2013 and summarize their chemical structures and bioactivities, aiming to lay a foundation for further development and utilization of these compounds and provide new ideas for the development of clinical drugs.

Research progress on Rhododendron molle in treatment of rheumatoid arthritis / 中国中药杂志

Rheumatoid arthritis(RA), as a chronic autoimmune disease, has a high incidence and disability rate, causing significant suffering to patients. Due to its complex pathogenesis, it has not been fully elucidated to date, and its treatment remains a challenging problem in the medical field. Although western medicine treatment options have certain efficacy, they require prolonged use and are expensive. Additionally, they carry risks of multiple infections and adverse reactions like malignancies. The Chinese herbal medicine Rhododendron molle is commonly used in folk medicine for its properties of dispelling wind, removing dampness, calming nerves, and alleviating pain in the treatment of diseases like rheumatic bone diseases. In recent years, modern clinical and pharmacological studies have shown that the diterpenoids in R. molle are effective components, exhibiting immune-regulatory, anti-inflammatory, and analgesic effects. This makes it a promising candidate for treating RA with a broad range of potential applications. However, R. molle has certain toxic properties that hinder its clinical application and lead to the wastage of its resources. This study reviewed recent research progress on the mechanism of R. molle in preventing and treating RA, focusing on its chemical components, anti-inflammatory and analgesic properties and summarized the adverse reactions associated with R. molle, aiming to offer new ideas for finding natural remedies for RA and methods to reduce toxicity while enhancing the effectiveness of R. molle. The study seeks to clarify the safety and efficacy of R. molle and its extracts, providing a theoretical basis for its application prospects and further promoting the development and utilization of R. molle resources.

Metabolism of four diterpenoids of Euphorbia fischeriana in liver microsomes of rats / 中国药房

OBJECTIVE To study the metabolites of four diterpenoids of Euphorbia fischeriana in liver microsomes of rats and to investigate its metabolic regularity. METHODS In vitro incubation system of liver microsomes of rats was built. The jolkinolide A，jolkinolide B ，17-hydroxyl jolkinolide A and 17-hydroxyl jolkinolide B were added into incubation system of liver microsomes in rats activated by reduced nicotinamide adenine dinucleotide phosphate ，incubated at 37 ℃ for 30 min，and then terminated the reaction with acetonitrile. Taking the negative group （adding acetonitrile firstly and then starting incubation for 30 min）as the reference，the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used ；Anaylyst®TF 1.7.1、PeakView® 2.2，MetabolitePilot 1.5 and MasterView 1.2 software were used to speculate and identify the fragmentation law of mass spectrometry and metabolites. RESULTS Four diterpenoids were easy to lose neutral fragments such as H 2O and CO in secondary mass spectrometry. Jolkinolide A and 17-hydroxyl jolkinolide A showed similar metabolism pathway ，including dihydroxylation，dehydrogenation，and monohydroxylation ；six and five metabolites were identified respectively. Jolkinolide B and 17-hydroxyl jolkinolide B showed similar metabolism pathway ，including monohydroxylation ，hydration and isomerization. Five metabolites were identified. CONCLUSIONS Both jolkinolide A and 17-hydroxyl jolkinolide A produce the metabolites of hydroxylation and dehydrogenation in liver microsomes of rats ；both jolkinolide B and 17-hydroxyl jolkinolide B produce the metabolites of hydroxylation ，hydration and isomerization in liver microsomes of rats. The metabolites of four diterpenoids are phase Ⅰ metabolites.

Cost-utility analysis of pertuzumab combined with trastuzumab and docetaxel in first-line treatment of HER 2- positive metastatic breast cancer / 中国药房

OBJECTIVE To study the metabolites of four diterpenoids of Euphorbia fischeriana in liver microsomes of rats and to investigate its metabolic regularity. METHODS In vitro incubation system of liver microsomes of rats was built. The jolkinolide A，jolkinolide B ，17-hydroxyl jolkinolide A and 17-hydroxyl jolkinolide B were added into incubation system of liver microsomes in rats activated by reduced nicotinamide adenine dinucleotide phosphate ，incubated at 37 ℃ for 30 min，and then terminated the reaction with acetonitrile. Taking the negative group （adding acetonitrile firstly and then starting incubation for 30 min）as the reference，the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used ；Anaylyst®TF 1.7.1、PeakView® 2.2，MetabolitePilot 1.5 and MasterView 1.2 software were used to speculate and identify the fragmentation law of mass spectrometry and metabolites. RESULTS Four diterpenoids were easy to lose neutral fragments such as H 2O and CO in secondary mass spectrometry. Jolkinolide A and 17-hydroxyl jolkinolide A showed similar metabolism pathway ，including dihydroxylation，dehydrogenation，and monohydroxylation ；six and five metabolites were identified respectively. Jolkinolide B and 17-hydroxyl jolkinolide B showed similar metabolism pathway ，including monohydroxylation ，hydration and isomerization. Five metabolites were identified. CONCLUSIONS Both jolkinolide A and 17-hydroxyl jolkinolide A produce the metabolites of hydroxylation and dehydrogenation in liver microsomes of rats ；both jolkinolide B and 17-hydroxyl jolkinolide B produce the metabolites of hydroxylation ，hydration and isomerization in liver microsomes of rats. The metabolites of four diterpenoids are phase Ⅰ metabolites.

Two new labdane diterpenoids from the leaves of Callicarpa formosana Rolfe / 药学学报

Two new labdane diterpenoids were isolated from 95% ethanol extract of the leaves of Callicarpa formosana Rolfe by using silica gel column, MCI column, ODS column and HPLC. Their structures were elucidated by HR-ESI-MS, NMR and ECD spectral data. All of them are new compounds, named 13E-6β-hydroxylabda-8(17),13-dien-15-oic acid (1) and 13E-7α-hydroxylabda-8(17),13-dien-15-oic acid (2). Compounds 1 and 2 were tested for antioxidant activity, and none of them had obvious activity.

A new labdane-type diterpenoids from Callicarpa nudiflora / 中国中药杂志

The purpose of the research is to study the bioactive constituents of Callicarpa nudiflora. From the 65% ethanol extract of C. nudiflora leaves, ten compounds were isolated by macroporous adsorption resin, Sephadex LH-20, ODS, silica gel, and preparative HPLC. These compounds were identified as callicapene M6(1), sterebin A(2), isomartynoside(3), crenatoside(4), luteolin-7-O-neohesperidoside(5), apigenin-7-O-β-D-neohesperidoside(6), isoacteoside(7), acteoside(8),(7R)-campneoside I(9), and(7S)-campneoside I(10) on the basis of NMR, HR-ESI-MS, and optical rotation data. Compound 1 was obtained as a new compound. Compounds 2 and 4 were isolated from the genus Callicarpa for the first time. Compounds 9 and 10 were isolated from C. nudiflora for the first time.

Research progress on chemical constituents from Chloranthus plants and their biological activities / 中国中药杂志

The genus Chloranthus has 13 species and 5 varieties in China, which can be found in the southwest and northeast regions. Phytochemical studies on Chloranthus plants have reported a large amount of terpenoids, such as diterpenoids, sesquiterpenoids, and sesquiterpenoid dimers. Their anti-inflammation, anti-tumor, antifungal, antivirus, and neuroprotection activities have been confirmed by previous pharmacological research. Herein, research on the chemical constituents from Chloranthus plants and their biological activities over the five years was summarized to provide scientific basis for the further development and utilization of Chloranthus plants.

A review: biosynthesis of plant-derived labdane-related diterpenoids / 中国天然药物

Plant-derived labdane-related diterpenoids (LRDs) represent a large group of terpenoids. LRDs possess either a labdane-type bicyclic core structure or more complex ring systems derived from labdane-type skeletons, such as abietane, pimarane, kaurane, etc. Due to their various pharmaceutical activities and unique properties, many of LRDs have been widely used in pharmaceutical, food and perfume industries. Biosynthesis of various LRDs has been extensively studied, leading to characterization of a large number of new biosynthetic enzymes. The biosynthetic pathways of important LRDs and the relevant enzymes (especially diterpene synthases and cytochrome P450 enzymes) were summarized in this review.

JYD01, a newly synthesized diterpenoid derivative, induces apoptosis in human gastric cancer cells through detaching hexokinase II from mitochondria / 中国药理学通报

Aim To investigate the effect of JYD01, an ent-kaurane diterpenoid analog, on detaching hexokinase II (HK II) from mitochondria, and discuss the underlying mechanism of anti-gastric cancer cell proliferation. Methods MTT assay was performed to measure the effect of JYD01 on the growth capacity of human gastric cancer cell lines MGC-803 and BGC-823. The glycolysis of MGC-803 cells in response to JYD01 was analyzed using a Seahorse XFp extracellular flux analyzer by real-time measurements of the extracellular acidification rate (ECAR, indicative of glycolysis). The effect of JYD01 in mitochondrial membrane potential (MMP) and apoptosis was observed by a fluorescence microscopy. The apoptotic rate and the quantitative analysis of MMP falling of cell lines treated with JYD01 were analyzed by flow cytometry. The proteins were determined by Western blot. Results JYD01 observably inhibited the growth of MGC-803 and BGC-823 cells in a dose-dependent manner. JYD01 induced a dose-dependent detachment of HK II from mitochondria of MGC-803 cells, effectively reduced glycolysis, and caused the drop of MMP leading to the release of cytochrome c. 1, 2 and 4 μmol · L

Taxane diterpenoids from the leaves and stems of Taxus wallichiana var. mairei / 中国药科大学学报

@#The chemical constituents of the CHCl3 extracts from the leaves and stems of Taxus wallichiana var. mairei were investigated. Twelve taxane diterpenoids were isolated and identified by spectroscopic analysis as 2-deacetoxytaxinine E (1),2-deacetoxytaxinine J (2),7-deacetoxytaxinine J (3),taxinine J (4),7,2′-didesacetoxyaustrospicatine (5),N-methyltaxol C (6),2-deacetoxydecinnamoyl taxinine J (7),taxol (8),7-epi-taxol (9),7-epi-10- deacetoxytaxol (10),cephalomannine (11),7-epi-cephalomannine (12). Compounds 1 and 3 were isolated from the leaves and stems of Taxus wallichiana var. mairei for the first time.

Study on Chemical Constituents of Diterpenoids from Buyi Medicine Isodon coetsa / 中国药房

OBJECTIVE：To st udy the chemical constituents of diterpenoids from Buyi medicine Isodon coetsa ，and to provide reference for the development and utilization of the medicinal resources. METHODS ：The 95％ methanol extract of Buyi medicine I. coetsa were isolated and purified with silica gel column ，Sehadex LH- 20 gel column and MCI column. The structures of the compounds were obtained by spectral analysis （mass spectrum ，hydrogen spectrum and carbon spectrum ），and then compared with active components of Miao medicine “Isodon flavidus ”. RESULTS & CONCLUSIONS ：Ten diterpenoids were obtained from I. coetsa，including rabdocoetsin B （Ⅰ），megathgrin B （Ⅱ），rabdocoetsin A （Ⅲ），enanderianin N （Ⅳ），rabdocoetsin D （Ⅴ）， megathyrin A （Ⅵ），lophanic acid （Ⅶ），rubesanolide D （Ⅷ），excisanin D （Ⅸ），excisanin K （Ⅹ）. The compounds Ⅶ，Ⅸ and Ⅹ were isolated from this specie for the first time. Compound Ⅶ（lophanic acid ）is a common active component in the Buyi medicine I. coetsa and Miao medicine “I. flavidus ”.

Anti-tumor Constituents from Rhizome of Euphorbia fischeriana / 中国实验方剂学杂志

Objective::To screen out active fraction from Euphorbia fischeriana, separate active components from E. fischeriana and explore structure-activity relationships, in order to analyze and identify chemical compositions of petroleum ether fraction from E. fischerian ethanol extract by ultra-performance liquid chromatography with quadrupole-time-of flight mass spectrometry (UPLC-Q-TOF-MS). Method::The anti-tumor activities of petroleum ether, ethyl acetate, n-butanol and water extracts from E. fischeriana were tested by methyl thiazolyl tetrazolium(MTT)method. A variety of modern chromatographic separation methods were used to separate active compounds from petroleum ether layer. Compounds were isolated. Their structures were identified by NMR technique. The structure-activity relationships between anti-tumor activities and structures of compounds were investigated. UPLC-Q-TOF-MS technique was used to identify the structures of petroleum ether extract from E. fischeriana. Mass spectrometry was performed in the positive ion mode using ESI ion sources. Result::Six compounds were isolated from petroleum ether fraction. They were jolkinolide A, jolkinolide B, 17-hydroxyjolkinolide A, 17-hydroxyjolkinolide B, euphopilolide and atis-16-en-13(s)-hydroxy-3, 14-dione. A total of 23 peaks were identified based on the comparison of retention times, accurate masses and fragmentation patterns with available standard compounds and literatures. Among them, there were 19 diterpenoids, 2 polyphenols, 1 fatty acid and 1 triterpenoid. Peaks No.18 and No.21 were tentatively identified as new compounds. Conclusion::The petroleum ether fraction showed a potential anti-tumor activity. The structure-activity relationships were discussed. UPLC-Q-TOF-MS technology can be used to quickly and accurately identify the structures, so as to provide a reference for its quality evaluation and active ingredient research.

Chemical constituents from Isodon amethystoides distributed in Libo / 中草药

Objective: To investigate the chemical constituents from Isodon amethystoides distributed in Libo. Methods: Chemical constituents were isolated and purified by chromatography with silica gel, MCI gel CHP 20P, and Sephadex LH-20. The structures of all compounds were elucidated by physicochemical properties, comprehensive spectral data and references. Results: Eighteen known compounds were isolated from the methanol extract of I. amethystoides, which were elucidated as lophanic acid (1), rubesanolide D (2), fladin A (3), 3,4-secoisopimara-4(18),7,15-triene-3-oic acid methylester (4), 3,4-secoisopimara-4(18),7, 15-triene-3-oic acid (5), isopimara-7,15-dien-3-one (6), isopimara-7,15-dien-3β-ol (7), alpiniol (8), (-)-clovane-2,9-diol (9), ursolic acid (10), oleanolic acid (11), epimaslinic acid (12), 7α-hydroxysitosterol (13), β-sitosterol (14), stigmasterol (15), sesamin (16), paulownin acetate (17) and dibutyl phthalate (18). Conclusion: Compound 4 is a new natural product, and compounds 2-9, 15-18 are isolated from I. amethystoides for the first time.

Study on chemical constituents of Abies chensiensis / 中草药

Objective: To study the chemical constituents of the sterns and leaves parts of Abies chensiensis. Methods: The isolation and purification were carried out by HP-20 macroporous resin, Sephadex LH-20, silica gel column chromatography, semi-preparative HPLC. Their structures were elucidated on the basis of physicochemical properties and spectroscopic data. Results: Twenty-four compounds were isolated and elucidated from the sterns and leaves parts of A. chensiensis, their structures were identified as 7,14,24-mariesatrien-26,23-olide-3α,23-diol (1), (23R)-3α-hydroxy-9,19-cyclo-9β-lanost-24-en-26,23-olide (2), 7-oxocallitrisic acid (3), 23-hydroxy-3-oxomariesia-8 (9),14,24-trien-26,23-olide (4), neoabiestrine E (5), 3-oxo-9β-lanosta-7,24-dien-26,23R-olide (6), 7,14,22Z,24-mariesatetraen-26,23-olide-3-one (7), (+)-rel-3α-hydroxy-23-oxocycloart-25 (27)-en-26-oic acid (8), cycloart-25-en- 3β,24-diol (9), neoabiestrine H (10), 3-oxo-24,25,26,27-tetranolanost-8-en-23-oic acid (11), abiesadine C (12), 13β-epidioxy-8 (14)-abieten-18-oic acid (13), dehydroabietic acid (14), 15-hydroxydehydroabietic acid (15), methyl 18-methoxydehydroabietate (16), 7β-hydroxy dehydroabietic acid (17), dehydroabietan-18-ol (18), centdaroic acid (19), abietic acid (20), 6,8,11,13-abi-etatrien-18-oic acid (21), abiesadine B (22), abiesadine N (23), 12β-hydroxyabietic acid (24). Conclusion: All compounds are isolated from the plants of A. chensiensis for the first time, and compound 11 is isolated from the Abies genus for the first time.