Identification and Molecular Characterization of the Alkaloid Biosynthesis Gene Family in Dendrobium catenatum

As one of the main active components of Dendrobium catenatum, alkaloids have high medicinal value. The physicochemicalproperties, conserved domains and motifs, phylogenetic analysis, and cis-acting elements of the genefamily members in the alkaloid biosynthesis pathway of D. catenatum were analyzed by bioinformatics, and theexpression of the genes in different years and tissues was analyzed by qRT-PCR. There are 16 gene families,including 25 genes, in the D. catenatum alkaloid biosynthesis pathway. The analysis of conserved domains andmotifs showed that the types, quantities, and orders of domains and motifs were similar among members ofthe same family, but there were significant differences among families. Phylogenetic analysis indicated that thegene family members showed some evolutionary conservation. Cis-acting element analysis revealed that therewere a large number of light-responsive elements and MYB (v-myb avian myeloblastosis viral oncogene homolog)-related elements in these genes. qRT-PCR showed that expressions of gene family members involved in alkaloidsynthesis were different in different years and tissues of D. catenatum. This study provides a theoretical basisfor further exploration of the regulatory mechanisms of these genes in the alkaloid biosynthesis of D. catenatum.

Deeply functional identification of TCS1 alleles provides efficient technical paths for low-caffeine breeding of tea plants

Caffeine is an important functional component in tea,which has the effect of excitement and nerve stimulation,but excessive intake can cause insomnia and dysphoria.Therefore,the production of tea with low-caffeine content can meet the consumption needs of certain people.Here,in addition to the previous alleles of the tea caffeine synthase(TCS1)gene,a new allele(TCS1h)from tea germplasms was identified.Results of in vitro activity analysis showed that TCS1h had both theobromine synthase(TS)and caffeine synthase(CS)activities.Site-directed mutagenesis experiments of TCS1a,TCS1c,and TCS1h demonstrated that apart from the 225th amino acid residue,the 269th amino acid also determined the CS activity.GUS histochemical analysis and dual-luciferase assay indicated the low promoter activity of TCS1e and TCS1f.In parallel,insertion and deletion mutations in large fragments of alleles and experiments of site-directed mutagenesis identified a key cis-acting element(G-box).Furthermore,it was found that the contents of purine alkaloids were related to the expression of corresponding functional genes and alleles,and the absence or presence and level of gene expression determined the content of purine alkaloids in tea plants to a certain extent.In summary,we concluded TCS1 alleles into three types with different functions and proposed a strategy to effectively enhance low-caffeine tea germplasms in breeding practices.This research provided an applicable technical avenue for accelerating the cultivation of specific low-caffeine tea plants.

New Neuraminidase Inhibitory Alkaloids from the Mangrove Soil-Derived Fungus Arthrinium sp.SCSIO 41305

New alkaloid,(E)-2-(hydroxyimino)-4-methylpentanamide(1)and a new cyclopentano[b]pyridine,4-hydroxy-7-methyl-6,7-dihydro-5H-cyclopenta[c]pyridin-5-one(2),together with ten known compounds(3–12)were isolated from the mangrove soil-derived fungus Arthrinium sp.SCSIO 41305.Extensive spectroscopic analysis and X-Ray crystallographic analysis were used to elucidate the structure of(E)-2-(hydroxyimino)-4-methylpentanamide(1),including its absolute configuration.All the isolated compounds(1–12)were evaluated for their antimicrobial and enzyme inhibitory activities against acetylcholinesterase(ACh E),neuraminidase(NAs),and phosphatidylinositol 3-kinase(PI3K).Among them,compounds 1 and 3 exhibited strong neuraminidase inhibitory activity with IC_(50)values of 12.04,1.92μmol L^(-1)(IC_(50)20μmol L^(-1)for oseltamivir acid),while compounds 5,6,8,and 10showed moderate neuraminidase inhibitory activity,and compounds 6–10 displayed weak enzyme inhibitory activities against PI3K.

Metabolic and transcriptomic analyses elucidate a novel insight into the network for biosynthesis of carbohydrate and secondary metabolites in the stems of a medicinal orchid Dendrobium nobile

Dendrobium nobile is an important medicinal and nutraceutical herb.Although the ingredients of D.nobile have been identified as polysaccharides,alkaloids,amino acids,flavonoids and bibenzyls,our understanding of the metabolic pathways that regulate the synthesis of these compounds is limited.Here,we used transcriptomic and metabolic analyses to elucidate the genes and metabolites involved in the biosynthesis of carbohydrate and several secondary metabolites in the stems of D.nobile.A total of 1005 metabolites and 31,745 genes were detected in the stems of D.nobile.The majority of these metabolites and genes were involved in the metabolism of carbohydrates(fructose,mannose,glucose,xylulose and starch),while some were involved in the metabolism of secondary metabolites(alkaloids,β-tyrosine,ferulic acid,4-hydroxybenzoate and chrysin).Our predicted regulatory network indicated that five genes(AROG,PYK,DXS,ACEE and HMGCR) might play vital roles in the transition from carbohydrate to alkaloid synthesis.Correlation analysis identified that six genes(ALDO,PMM,BGLX,EGLC,XYLB and GLGA) were involved in carbohydrate metabolism,and two genes(ADT and CYP73A) were involved in secondary metabolite biosynthesis.Our analyses also indicated that phosphoenol-pyruvate(PEP) was a crucial bridge that connected carbohydrate to alkaloid biosynthesis.The regulatory network between carbohydrate and secondary metabolite biosynthesis established will provide important insights into the regulation of metabolites and biological systems in Dendrobium species.

Design,Synthesis and Bioactivity Study of Marine Alkaloid Neobacillamide-A Derivatives

The Janus kinases(JAKs)are a family of intracellular tyrosine kinases that play an essential role in many basic biological processes,such as apoptosis and inflammation.Thus any dysfunction of the proteins in this pathway may lead to a variety of diseases,including cancer and diseases that affect the immune system,such as severe combined immune deficient(SCID).Marine biological resources have become an important source in new drug research and development due to their diversity,complexity and speciality.In this study,Marine alkaloid Neobacillamide A was isolated from the greedy and stubborn sponge symbiotic Bacillus atrophicus C89 in the South China Sea.Totally 24 novel marine alkaloid Neobacillamide A derivatives were designed and synthesized,which were evaluated for their inhibitory activity against JAK/STAT signaling pathway and their cytotoxicity to A549 cells.Compounds 13c,13o,14d,14g and 14h showed potent JAK/STAT inhibition capability(concentration of 25μmol L^(-1),all inhibitory potencies were above 60%),especially compound 14g exhibited superior JAK/STAT inhibition effect(89.70%inhibition).In addition,all these compounds with a concentration of 25μmol L^(-1)displayed weak or no cytotoxicity to A549 cells,which means that these Neobacillamide A derivatives with JAK/STAT inhibition capability may have potential anti-inflammatory function.

Discovery of prenylated indole alkaloid and natural xanthone from cold-seep sediment derived fungus Talaromyces funiculosus SD-523

A new prenylated indole alkaloid 11,17-epi-mangrovamide A(1),a new natural occurring product,1,7-dihydroxy-6-methyl-8-hydroxymethyl-xanthone(2),two known alkaloids,mangrovamide A(3)and mangrovamide G(4),and four known polyketide derivatives(5–8)were isolated and identified from the cold-seep sediment derived fungal strain Talaromyces funiculosus SD-523.Their structures were elucidated by combination of nuclear magnetic resonance(NMR),high resolution electrospray ionization mass spectroscopy(HRESIMS),quantum chemical electronic circular dichroism(ECD),and DP4+probability analysis as well as by comparison of the data with literature reports.All isolated compounds were tested for antibacterial activities.

A Pair of New Spirocyclic Alkaloid Enantiomers with TrxR Inhibitory Activities Were Isolated from Marine-Derived Aspergillus ruber TX-M4-1

One new spirocyclic alkaloid,5-isopentenyl-cryptoechinuline D(1),along with 11 known compounds(2–12),were iso-lated from a marine fungus Aspergillus ruber TX-M4-1.The structures of compounds 1–12 were elucidated by spectroscopic evi-dences.Compound 1 was initially isolated as an enantiomer,and further separation of 1 by chiral HPLC afforded a pair of enantio-mers,including(-)-5-isopentenyl-cryptoechinuline D(1a)and(+)-5-isopentenyl-cryptoechinuline D(1b).Their absolute configura-tions were elucidated by ECD spectroscopic data.Compounds 1a,5 and 10 could inhibit thioredoxin reductase(TrxR)activity with IC50 values of 6.2,36.3 and 18.6μmol L^(-1),respectively.Surface plasmon resonance(SPR)study also demonsrated the interactions between compounds 6,8 and Niemann-Pick C1 Like 1(NPC1L1)respectively,which indicate that compounds 6 and 8 are potential NPC1L1 inhibitors.

Biological Activity and Molecular Mechanism of Bullatine G

Bullatine G has many biological effects such as anti-inflammatory,anti-anxiety,anti-tumor,anti-arrhythmia and anti-heart failure effect.In order to provide a theoretical basis for the further study and clinical application of bullatine G,this article reviews the biological activity of bullatine G in recent years.

Effects of Different Alkaloids in Coptis chinensis on Inhibiting TGEV Proliferation

[Objectives]To study the effects of different alkaloids in Coptis chinensis on inhibiting the proliferation of Transmissible gastroenteritis virus(TGEV).[Methods]The components and content of the main alkaloids in the extract of C.chinensis were analyzed.The main alkaloids were selected as drugs to inhibit the proliferation of TGEV.The maximum non-toxic concentration of Columbamine,Jatrorrhizine,Epiberberine,Coptisine,Palmatine,and Berberine was screened.The protective rate of each drug on TGEV-infected ST cells was determined,and the transcriptional inhibitory effect of the drug on TGEV N gene was detected by fluorescent quantitative PCR.[Results]The extract of C.chinensis mainly contains 6 alkaloids:Columbamine,Jatrorrhizine,Epiberberine,Coptisine,Palmatine,and Berberine,accounting for 2.03%,8.88%,9.21%,15.07%,14.63%,and 50.18%,respectively.In the range of the safe concentration,Jatrorrhizine,Palmatine,and Coptisine had better protective effects on ST cells infected with TGEV;compared with the Columbamine group,the cell protection rate was significantly different(P<0.05);compared with the Berberine group,the difference was extremely significant(P<0.01).The Coptisine and Palmatine groups had significant inhibitory effects on the transcription of TGEV N gene,and the difference was extremely significant compared with the virus group(P<0.05).[Conclusions]Jatrorrhizine and Palmatine in C.chinensis are the main components to inhibit the proliferation of TGEV.

Phytochemical Screening of Some Medicinal Plants in Al Jouf, KSA

The utilization of ethnobotanical and phytochemical investigations in the discovery of novel medications is beneficial. Screening for phytochemicals is an important step in detecting the bioactive ingredients of medicinal plants which are used in conventional therapy. For the first time, 23 medicinal plants utilized in Saudi Arabian traditional therapy were examined. From August 2020 to July 2021, ethnobotanical fieldwork was conducted. There was some plant species identified, divided into pertinent families. Standard procedures were used to screen these medicinal plants for the occurrence of glycosides, alkaloids, saponins, resins, saponins, tannins, and flavonoids. Among the medicinal plants used, the most common phytochemicals were alkaloids (95.65%), glycosides (86.96%), saponin (82.61%), tannins (73.91%), flavonoids (56.52%), and resin (52.17%). The least widely distributed chemicals, on the other side, were resins. Trigonella foenum-graecum L., Pimpinella anisum L., and Cuminum cyminum L. seeds were shown to contain all six categories of secondary metabolites. The ethnographic importance of these medicinal plants is consistent with the content of secondary metabolites.

微波加工对附子中酯型生物碱的影响

[目的]探讨微波加工对附子中双、单酯型生物碱含量的影响,为附子新型炮制品及药食两用加工品的开发提供参考。[方法]通过正交设计、直观分析和方差分析,探讨烫漂时间(A)、干燥温度(B)和微波时间(C)对附子中双、单酯型生物碱含量的影响。[结果]微波时间对双酯型生物碱含量具有极显著影响(P<0.01),对单酯型生物碱含量具有显著影响(P<0.05);烫漂时间和干燥温度仅对双酯型生物碱含量具有显著影响(P<0.05)。最佳的因素水平组合为A3B3C3,即烫漂时间为3 min,干燥温度为60℃,微波时间为5 min;在此条件下,双酯型生物碱含量为590.33μg/g,单酯型生物碱含量为802.14μg/g。[结论]微波加工能够降低附子中双酯型生物碱含量,增加单酯型生物碱含量。

苦地丁总生物碱对小鼠Ⅱ型胶原关节炎的影响

目的探讨苦地丁对小鼠Ⅱ型胶原关节炎的影响。方法小鼠尾根部皮内注射Ⅱ型胶原乳剂建立小鼠关节炎模型(CIA),苦地丁总生物碱(50,100,200mg·kg-1组灌胃给药,每日1次;模型组和空白组灌胃给予等量蒸馏水;实验结束时考察各组临床积分、发病时间、血清中抗-CⅡ抗体含量、胶原所致耳廓DTH;ELISA法检测PGE2、TNF-α、IL-1的水平。结果苦地丁剂量依赖性延迟CIA小鼠的发病时间,减少关节的肿胀和红斑,缓解CIA小鼠体质量减轻,降低小鼠血清中抗-CⅡ抗体水平,抑制CⅡ诱导的小鼠耳廓迟发型超敏反应;明显抑制巨噬细胞释放肿瘤坏死因子(TNF-α)、白介素1(IL-1),但对前列腺素(PGE2)的产生无明显影响。结论苦地丁对小鼠CIA有明显的抑制作用。

土豆中配糖碱的测定(综述)

在土豆中存在的主要配糖碱(glyco-alkaloid)有α—茄碱(α—solanine)和α—卡茄碱(α—Chaconine)及其代谢物(图1),其中前二者占95～99%。配糖碱具有特殊的苦味,其含量与土豆品种,栽培条件及收获后处理与贮存有关,而且,物理或化学的代射抑制、光照射等都可能产生配糖碱。这些配糖碱具有急性毒性和可能的慢性毒性,现普遍认为。

不同理化因子对黄连愈伤组织生长和生物碱合成的影响

不同理化因子对黄连愈伤组织生长和生物碱合成的影响张浩１陈炜晁若冰王雪耕２庄燕黎２李小华２（华西医科大学药学院成都６１００４１）古蔺野连（又称串珠连Ｃｏｐｔｉｓｇｕｌｉｎｅｎｓｉｓ）为黄连属植物中的一个野生珍稀品种，只生长在我国西南少数山区，其根茎可作...

A review on medicinal importance,pharmacological activity and bioanalytical aspects of beta-carboline alkaloid "Harmine"

Harmine,a beta-carboline alkaloid,is widely distributed in the plants,marine creatures,insects, mammalians as well as in human tissues and body fluids.Harmine was originally isolated from seeds of Peganum harmal in 1847 having a core indole structure and a pyridine ring.Harmine has various types of pharmacological activities such as antimicrobial,antifungal,antitumor,cytotoxic, antiplasmodial,antioxidaant,antimutagenic,antigenotoxic and hallucinogenic properties.It acts on gamma-aminobutyric acid type A and monoamine oxidase A or B receptor,enhances insulin sensitivity and also produces vasorelaxant effect.Harmine prevents bone loss by suppressing osteoclastogenesis.The current review gives an overview on pharmacological activity and analytical techniques of harmine,which may be useful for researcheres to explore the hidden potential of harmine and and will also help in developing new drugs for the treatment of various diseases.

Pyrrolizidine alkaloids-induced hepatic sinusoidal obstruction syndrome: Pathogenesis, clinical manifestations, diagnosis,treatment, and outcomes

Hepatic sinusoidal obstruction syndrome (HSOS) can be caused by the intake of pyrrolizidine alkaloids (PAs). To date, PAs-induced HSOS has not been extensively studied. In view of the difference in etiology of HSOS between the West and China, clinical profiles, imaging findings, treatment, and outcomes of HSOS associated with hematopoietic stem cell transplantation or oxaliplatin might be hardly extrapolated to PAs-induced HSOS. Reactive metabolites derived from PAs form pyrrole-protein adducts that result in toxic destruction of hepatic sinusoidal endothelial cells. PAs-induced HSOS typically manifests as painful hepatomegaly, ascites, and jaundice. Laboratory tests revealed abnormal liver function tests were observed in most of the patients with PAs-induced HSOS. In addition, contrast computed tomography and magnetic resonance imaging scan show that patients with PAs-induced HSOS have distinct imaging features, which reveal that radiological imaging provides an effective noninvasive method for the diagnosis of PAs-induced HSOS. Liver biopsy and histological examination showed that PAs-induced HSOS displayed distinct features in acute and chronic stages. Therapeutic strategies for PAs-induced HSOS include rigorous fluid management, anticoagulant therapy, glucocorticoids, transjugular intrahepatic portosystemic shunt, liver transplantation, etc. The aim of this review is to describe the pathogenesis, clinical profiles, diagnostic criteria, treatment, and outcomes of PAs-induced HSOS.

Qualitative and Quantitative Analysis of Alkaloids in Cortex Phellodendri by HPLC-ESI-MS/MS and HPLC-DAD

A combined method of high performance liquid chromatograph-elecrtrospray-ionization mass spectrometer（HPLC-ESI-MS/MS） coupled with a photodiode array detector（HPLC-DAD） and principal component analysis（PCA） was applied to the qualitative and quantitative analyses of alkaloids in Cortex Phellodendri（CP） samples, and to the differentiation of two species of CP, Cortex Phellodendri Chinensis（CPC） and Cortex Phellodendri Amurensis（CPA）. Twenty-two peaks appeared in the HPLC-MS base peak chromatogram of CP detected by the HPLC-ESI-MS/MS analysis, and the alkaloids were identified according to the MSn data, the known MS fragmentation rules and the literature data. Five alkaloids including berberine, palmatine, jatrorrhizine, phellodendrine and magnoflorine were simultaneously determinated by the HPLC-DAD. Berberine was the primary component in all CP samples, and the contents of berberine and palmatine were exploited to be two critical parameters for effective discrimination between the two species of CP. The average content of berberine in CPC（58.75 mg/g） was higher than that in CPA（9.16 mg/g）, while the content of palmatine was less, only 0.25 mg/g in CPC and 4.19 mg/g in CPA. With the use of PCA, samples datasets were separated successfully into two different clusters corresponding to the two species, and berberine, pahnatine, phellodendrine and magnoflorine contribute most to the above mentioned calssifying . The proposed method oroved to be a useful tool in the aualitv control of Chinese herbal medicines.

Effect of processing on the alkaloids in Aconitum tubers by HPLC-TOF/MS

According to the Chinese Pharmacopoeia 2015, only processed Aconitum tubers can be clinically applied, and the effect of processing is unclear. This research aimed to explore the effect of processing on cardiac efficacy of alkaloids in Aconitum tubers. First, the chemical ingredients in unprocessed and processed Aconitum tubers were identified and compared by using high performance liquid chromatography time-of-flight mass spectrometry(HPLC-TOF/MS) and multivariate pattern recognition methods. Then the representative alkaloids in Aconitum tubers, aconitine, benzoylaconine, and aconine, which belong to diester-diterpenoid alkaloids,monoester-diterpenoid alkaloids, and amine-diterpenoid alkaloids, respectively, were selected for further validation of attenuated mechanism. Subsequent pharmacological experiments with aconitine, benzoylaconine,and aconine in SD rats were used to validate the effect of processing on cardiac functions. After processing the Aconitum tubers, it was found that the contents of diester-diterpenoid alkaloids were reduced, and those of monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids were increased, suggesting that diesterditerpenoid alkaloids were transformed into monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids.Through further decocting the aconitine in boiling water, it was confirmed that the three alkaloids could be progressively transformed. Pharmacological experiments with aconitine, benzoylaconine, and aconine in SD rats showed that aconitine at a dose of 0.01 mg/kg and aconine at a dose of 10 mg/kg enhanced the cardiac function, while benzoylaconine at a dose of 2 mg/kg weakened the cardiac function. The effect of processing is attributed to the transformation of the most toxic diester-diterpenoid alkaloids into less toxic monoesterditerpenoid alkaloids and amine-diterpenoid alkaloids.

Hepatoprotective and antioxidant activity of a mangrove plant Lumnitzera racemosa

Objective:To identify the hepatoprotective and in vitro antioxidant activity of Lumnitzera racemosa(L.racemosa)leaf extract.Methods:Animals in Group 1 served as vehicle control,Group 2 served as liepatoloxin(CCL_4 treated)group,Group 3 served as positive control(Silyinarin)group,and Group 4,5 and 6 served as(75,150 and 300 mg/kg bw p.o.)L.raremosa leaf extract treated groups.Moreover,in vitro antioxidant DPPH,hydroxyl radical scavenging activity(HRSA),NO,ferric reducing antioxidant power(FRAP),lipid hydroperoxide(LPO)and super oxide dismutase(SOD)were also analyzed for the leaf extract.Results:The levels of the serum parameters such as serum glutamic oxaloacetic transaminase(SCOT),serum glutamic pyruvic transaminase(SGPT),alkaline phosphatase(ALP),bilirubin,cholesterol(CHL),sugar and lactate dehydrogenase(LDU)were significantly increased in CCL_4 treated rats when compared with the control group(P<0.05).But the L.racemosa leaf extract treated rats showed maximum reduction of SGOT[(210.36±19.63)IU/L],SGPT[(82.37±13.87)IU/L],ALP[(197.63±23.43)IU/L],bilurubin[(2.15±0.84)mg/dL],cholesterol[(163.83±15.63)mg/dL],sugar[(93.00±7.65)mg/dL]and LDH[(1134.00±285.00)IU/L]were observed with the high dose(300 mg/kg bw)of leaf extract treated rats.Histopathological scores showed that,no visible changes were observed with high dose(300 mg/kg bw)of leaf extract treated rats except few mild necrosis.The IC_(50)values were observed as(56.37±4.87)μg/mL,(57.68±1.98)μg/mL,(64.I5±2.90)μg/mL,(61.94±3.98)μg/mL,(94.53±1.68)μg/mL and(69.7±2.65)μg/mL for DPPH,HRSA,NO,FRAP,LPO and SOD radical scavenging activities,respectively.Conclusions:In conclusion,the hepatoprotective effect of the L.racemosa leaf extract might be due to the presence of phenolic groups,terpenoids and alkaloids and in vitro antioxidant properties.

A New Dendrobine-Type Alkaloid from Dendrobium nobile

A new dendrobine-type alkaloid has been isolated from stems of Dendrobium nobile lindl.. Its structure has been identified on the basis of spectroscopic method.