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Hyoscyamine (Hyos) and scopolamine (SCP) are drugs widely used as antimuscarinic to treat diseases such as Parkinson’s or to calm schizoid patients. In this study, with the aim of enhancing tropane...

N-methylputrescine is the precursor of nicotine and pharmaceutical tropane alkaloids such as hyoscyamine. Putrescine N-methyltransferase (PMT) catalyzes the N-methylation of putrescine to form N-methylputrescine. While the role of PMT in nicotine biosynthesis is clear, knowledge of PMT in the biosynthesis of tropane alkaloids (TAs) and the regulation of polyamines remains limited. We characterized a PMT gene from Hyoscyamus niger, designated HnPMT that was specifically expressed in roots, especially in the secondary roots and dramatically induced by methyl jasmonate (MeJA). The GUS gene was specifically expressed in Arabidopsis roots or in the vascular tissues, including pericycles and endodermis, of the H. niger hairy root cultures, when it was driven by the 5'-flanking promoter region of HnPMT. The recombinant HnPMT was purified for enzymatic assays. HnPMT converted putrescine to form N-methylputrescine, as confirmed by LC-MS. The kinetics analysis revealed that HnPMT had high affinity with putrescine but low catalytic activity, suggesting that it was a rate-limiting enzyme. When HnPMT was suppressed in the H. niger plants by using the VIGS approach, the contents of N-methylputrescine and hyoscyamine were markedly decreased, but the contents of putrescine, spermidine and a mixture of spermine and thermospermine were significantly increased; this suggested that HnPMT was involved in the biosynthesis of tropane alkaloids and played a competent role in regulating the biosynthesis of polyamines. Functional identification of HnPMT facilitated the understanding of TA biosynthesis and thus implied that the HnPMT-catalyzed step might be a target for metabolic engineering of the TA production in H. niger.

This Article describes the development of a second-generation catalyst system for the transannular C–H functionalization of alicyclic amines. Pyridine- and quinoline-carboxylate ligands are shown to be highly effective for increasing the reaction rate, yield, and scope of Pd-catalyzed transannular C–H arylation reactions of azabicyclo[3.1.0]hexane, azabicyclo[3.1.1]heptane, azabicyclo[3.2.1]octane, and piperidine derivatives. Mechanistic studies reveal that the pyridine/quinoline-carboxylates play a role in impeding both reversible and irreversible catalyst decomposition pathways. These ligands enable the first reported examples of the transannular C–H arylation of the ubiquitous tropane, 7-azanorbornane, and homotropane cores. Finally, the pyridine/quinoline-carboxylates are shown to promote both transannular C–H arylation and transannular C–H dehydrogenation on a homotropane substrate.

Scopolia lurida, a medicinal plant native to the Tibetan Plateau, is among the most effective producers of pharmaceutical tropane alkaloids (TAs). The hyoscyamine 6β-hydroxylase genes of Hyoscyamus niger (HnH6H) and S. lurida (SlH6H) were cloned and respectively overexpressed in hairy root cultures of S. lurida, to compare their effects on promoting the production of TAs, especially the high-value scopolamine. Root cultures with SlH6H/HnH6H overexpression were confirmed by PCR and real-time quantitative PCR, suggesting that the enzymatic steps defined by H6H were strongly elevated at the transcriptional level. Tropane alkaloids, including hyoscyamine, anisodamine and scopolamine, were analyzed by HPLC. Scopolamine and anisodamine contents were remarkably elevated in the root cultures overexpressing SlH6H/HnH6H, whereas that of hyoscyamine was more or less reduced, when compared with those of the control. These results also indicated that SlH6H and HnH6H promoted anisodamine production at similar levels in S. lurida root cultures. More importantly, HnH6H-overexpressing root cultures had more scopolamine in them that did SlH6H-overexpressing root cultures. This study not only provides a feasible way of overexpressing H6H to produce high-value scopolamine in engineered root cultures of S. lurida but also found that HnH6H was better than SlH6H for engineering scopolamine production.

Ionic liquids (ILs) have been widely used in the field of extraction of natural bioactive compounds because of their advantages compared to traditional organic solvents. In this study, the new ‘like dissolves like’ mode was designed and seven types of tropine-based ionic liquids were used to extract tropane alkaloids from radix physochlainae, and then the relationship between their performance and structures together with the effects of main extraction conditions were explored. It was found that 0.05 mol L−1 [C3tr][PF6] aqueous solution had the ideal selectivity and high extraction efficiency of 95.1% at 75 °C when the extraction time was 55 min and the solid–liquid ratio was 1 : 35, which was superior to that of 85% ethanol–water and 0.1% hydrochloric acid–water. There was no decomposition and racemization of products occurring in the mixture solution when above extraction solvent was applied. In addition, the extraction behavior and mechanism using an ionic liquid aqueous solution was tentatively studied through thermodynamics experiments, near-infrared/infrared spectroscopy (NIR/IR), scanning electron microscopy (SEM), and thermogravimetric analysis (TG), and subsequent back-extraction could be efficiently used to further separate alkaloids and ILs. In the developed ‘like dissolves like’ mode, the extraction process of target alkaloids was found to be endothermic and spontaneous through the specific interaction between them and the solvent molecules with the same nucleus.

The consumption of animal products is rapidly increasing and, therefore, the use of feedstuffs in farms is in demand in developing countries. Feed cereals represent more than 48% of total feed material and they might be contaminated by Solanaceae plants, which contain toxic metabolites such as tropane alkaloids. A wide-scope analysis of tropane alkaloids in feed, including atropine, scopolamine, tropinone, anisodamine, tropane, homatropine, tropine, littorine, physoperuvine, pseudotropine, apoatropine and aposcopolamine, has been performed using liquid chromatography coupled to an Exactive-Orbitrap analyzer. An extraction method based on pressurized liquid extraction with methanol/water/formic acid (75/25/0.4; v/v/v) as a solvent extraction, followed by a solid phase extraction step using Strata-X-C cartridges and a clean-up step with chitosan has been used. The method was fully validated and recoveries from 70 to 109% with an intraday precision lower than or equal to 15% were achieved and limits of quantification (LOQs) ranged from 5 to 25 μg kg−1 for all the compounds. More than 40 samples belonging to several classes of animal feed (cow, rabbit, sheep, chicken and pig) were analysed, showing the occurrence of tropane alkaloids in 18 out of 45 samples, with concentration ranging from 5 (atropine) to 222 μg kg−1 (scopolamine), highlighting the need of analytical methods for the control of TAs in animal feeds.

Compared with silver nanoparticles, gold nanoparticles have excellent chemical resistance and biocompatibility, while silver nanoparticles have more excellent local surface plasmon resonance characteristics than gold. In this article, we developed a simple method for growing island-shaped gold nanoparticles on the surface of silver nanowires (NWs), so that the prepared Ag@Au NWs have both the advantages of Au and Ag without their disadvantages. Furthermore, by changing the reaction parameters, we achieved the regulation of the density of island-shaped gold nanoparticles on the Ag NWs surface, thereby optimizing its LSPR performance. We use the prepared Ag@Au NWs as the base material for highly sensitive Surface enhanced Raman scattering (SERS) detection. By using 4-aminothiophenol (4-ATP) as the test molecule to explore the uniformity of the prepared SERS substrate, the detection limit of 4-ATP reached 10-9 M. And 10 points were randomly selected for testing and the RSD was 7.4%, indicating the Ag@Au NWs SERS substrates had excellent uniformity. Furthermore, we have achieved highly sensitive SERS detection of 5 tropane alkaloids including L -Hyoscyamine, Anisodamine, Scopolamine Hydrobromide, scopolamine methyl bromide and scopolamine butyrate bromide. The limit of detection is 10-9 M, 10-10 M, 10-9 M, 10-9 M and 10-10 M. Furthermore, by analyzing the obtained SERS spectra, we realized the differentiation of 5 kinds of tropane alkaloids by their characteristic peaks.

Atropa belladonna, one of the most important medicinal plants in China, is used to produce anticholinergic tropane alkaloids. Weeds severely reduce the agricultural productivity of A. belladonna in fields, and the low content of high-value scopolamine in planta limits its economic value. In this study, homozygous lines of A. belladonna, with glyphosate resistance and high yield of scopolamine, were developed using metabolic engineering technology for the first time. Two homozygous transgenic lines (T2GH03 and T2GH06), harboring G2-EPSPS and HnH6H, were established and tested in the field. Glyphosate resistance assays demonstrated that the two homozygous transgenic lines could resist commercial recommended dose of glyphosate. Alkaloid analysis showed that the transgenic lines produced more scopolamine than wild-type plants. T2GH03 and T2GH06 produced scopolamine at the levels of 5.45 mg/g dry weight (DW) and 7.04 mg/g DW respectively in leaves, and 1.06 mg/g DW to 1.07 mg/g DW respectively in roots. Compared with wild-type plants, the scopolamine contents of homozygous transgenic lines increased by over 10 folds in leaves and nearly 5 folds in roots. To sum up, Atropa belladonna’s new varieties with glyphosate resistance and high-yield scopolamine are developed, which are highly valuable for industrial production of scopolamine.

Curcumin (CUR) is a natural compound that exhibits anti-inflammatory, anti-bacterial, and other biological properties. However, its application as an effective drug is problematic due to its poor oral bioavailability, solubility in water, and poor absorption from the gastrointestinal tract. The aim of this work is to synthesize monocarbonyl analogs of CUR based on the 9-methyl-9-azabicyclo[3.2.1]nonan-3-one (pseudopelletierine, granatanone) scaffold to improve its bioavailability. Granatane is a homologue of tropane, whose structure is present in numerous naturally occurring alkaloids, e.g., l-cocaine and l-scopolamine. In this study, ten new pseudopelletierine-derived monocarbonyl analogs of CUR were successfully synthesized and characterized by spectral methods and X-ray crystallography. Additionally, in vitro test of the cytotoxicity and anti-inflammatory properties of the synthesized compounds were performed.

Transgenic hairy roots of Datura spp., established using strain A4 of Agrobacterium rhizogenes, are genetically stable and produce high levels of tropane alkaloids. To increase biomass and tropane alkaloid content of this plant tissue, four Pseudomonas strains, Pseudomonas fluorescens P64, P66, C7R12, and Pseudomonas putida PP01 were assayed as biotic elicitors on transgenic hairy roots of Datura stramonium, Datura tatula, and Datura innoxia. Alkaloids were extracted from dried biomass, and hyoscyamine and scopolamine were quantified using liquid chromatography-tandem mass spectrometry analysis. D. stramonium and D. innoxia biomass production was stimulated by all Pseudomonas spp. strains after a 5-d treatment. All strains of P. fluorescens increased hyoscyamine yields compared to untreated cultures after both 5 and 10 d of treatment. Hyoscyamine yields were highest in D. tatula cultures exposed to a 5-d treatment with C7R12 (16.633 + 0.456 mg g−1 dry weight, a 431% increase) although the highest yield increases compared to the control were observed in D. stramonium cultures exposed to strains P64 (511% increase) and C7R12 (583% increase) for 10 d. D. innoxia showed the highest scopolamine yields after elicitation with P. fluorescens strains P64 for 5 d (0.653 + 0.021 mg g−1 dry weight, a 265% increase) and P66 for 5 and 10 d (5 d, 0.754 + 0.0.031 mg g−1 dry weight, a 321% increase; 10 d 0.634 + 0.046 mg g−1 dry weight, a 277% increase). These results show that the Pseudomonas strains studied here can positively and significantly affect biomass and the yields of hyoscyamine and scopolamine from transgenic roots of the three Datura species.

Awareness about pyrrolizidine alkaloids (PAs) and tropane alkaloids (TAs) in food was recently raised by the European Food Safety Authority stressing the lack of data and gaps of knowledge required to improve the risk assessment strategy. The present study aimed at the elaboration and validation of a method to determine PAs and TAs in honey. QuEChERS sample treatment and liquid chromatography coupled to hybrid high resolution mass spectrometry, were used. The method resulted in good linearity (R 2 > 0.99) and low limits of detection and quantification, ranging from 0.04 to 0.2 µg kg −1 and from 0.1 to 0.7 µg kg −1 respectively. Recoveries ranged from 92.3 to 114.8% with repeatability lying between 0.9 and 15.1% and reproducibility between 1.1 and 15.6%. These performances demonstrate the selectivity and sensitivity of the method for simultaneous trace detection and quantification of PAs and TAs in honey, verified through the analysis of forty commercial samples.

The title compound is a salt containing a protonated endo-3-amino­tropane cage and a novel anionic copper(II) complex, [CuCl3(NO3)(H2O)]2−., The structure of a salt of diprotonated endo-3-amino­tropane crystallized with a copper(II) anionic cluster is reported, viz. (C8H18N2)[CuCl3(NO3)(H2O)]. Neither ion in the salt has been structurally characterized previously. In the crystal, the ions pack together to form a three-dimensional structure held together by a network of inter­molecular N—H⋯O, O—H⋯Cl and N—H⋯Cl hydrogen-bonding inter­actions. Selective crystallization of the title compound can be considered as a simple method for the separation of the exo and endo isomers of 3-amino­tropane.

Tropane alkaloids (TAs), especially hyoscyamine and scopolamine, are important precursors for anticholinergic and antispasmodic drugs. Hyoscyamine and scopolamine are currently obtained at commercial scale from hybrid crosses of Duboisia myoporoides × Duboisia leichhardtii plants. In this study, we present a global investigation of the localization and organization of TA biosynthesis in a Duboisia myoporoides R. Br. wild-type line. The tissue-specific spatial distribution of TAs within D. myoporoides is presented, including quantification of the TAs littorine, 6-hydroxy hyoscyamine, hyoscyamine, scopolamine and, additionally, hyoscyamine aldehyde as well as scopolamine glucoside. Scopolamine (14.77 ± 5.03 mg g-1), and to a lesser extent hyoscyamine (3.01 ± 1.54 mg g-1) as well as 6-hydroxy hyoscyamine (4.35 ± 1.18 mg g-1), are accumulated in leaves during plant development, with the highest concentration of total TAs detected in 6-month-old plants. Littorine, an early precursor in TA biosynthesis, was present only in the roots (0.46 ± 0.07 mg g-1). During development, the spatial distribution of all investigated alkaloids changed due to secondary growth in the roots. Transcripts of pmt, tr-I and cyp80f1 genes, involved in early stages of TA biosynthesis, were found to be most abundant in the roots. In contrast, the transcript encoding hyoscyamine 6β-hydroxylase (h6h) was highest in the leaves of 3-month-old plants. This investigation presents the spatial distribution of biochemical components as well as gene expression profiles of genetic factors known to participate in TA biosynthesis in D. myoporoides. The results of this investigation may aid in future breeding or genetic enhancement strategies aimed at increasing the yields of TAs in these medicinally valuable plant species.

Dopamine transporter (DAT) blockers like cocaine and many other abused and therapeutic drugs bind and stabilize an inactive form of the transporter inhibiting reuptake of extracellular dopamine (DA). The resulting increases in DA lead to the ability of these drugs to induce psychomotor alterations and addiction, but paradoxical findings in animal models indicate that not all DAT antagonists induce cocaine-like behavioral outcomes. How this occurs is not known, but one possibility is that uptake inhibitors may bind at multiple locations or in different poses to stabilize distinct conformational transporter states associated with differential neurochemical endpoints. Understanding the molecular mechanisms governing the pharmacological inhibition of DAT is therefore key for understanding the requisite interactions for behavioral modulation and addiction. Previously, we leveraged complementary computational docking, mutagenesis, peptide mapping, and substituted cysteine accessibility strategies to identify the specific adduction site and binding pose for the crosslinkable, photoactive cocaine analog, RTI 82, which contains a photoactive azide attached at the 2β position of the tropane pharmacophore. Here, we utilize similar methodology with a different cocaine analog N-[4-(4-azido-3-I-iodophenyl)- butyl]-2-carbomethoxy-3- (4-chlorophenyl) tropane, MFZ 2–24, where the photoactive azide is attached to the tropane nitrogen. In contrast to RTI 82, which crosslinked into residue Phe319 of transmembrane domain (TM) 6, our findings show that MFZ 2–24 adducts to Leu80 in TM1 with modeling and biochemical data indicating that MFZ 2–24, like RTI 82, occupies the central S1 binding pocket with the (+)-charged tropane ring nitrogen coordinating with the (−)-charged carboxyl side chain of Asp79. The superimposition of the tropane ring in the three-dimensional binding poses of these two distinct ligands provides strong experimental evidence for cocaine binding to DAT in the S1 site and the importance of the tropane moiety in competitive mechanisms of DA uptake inhibition. These findings set a structure-function baseline for comparison of typical and atypical DAT inhibitors and how their interactions with DAT could lead to the loss of cocaine-like behaviors.

Moallem E, Ghasemipirbalouti A, Nejadsattari T, Iranbakhsh A, Mehregan I. 2017. Evaluation of expression analysis of putrescine n-methyltransferase gene during different stages of growth in the medicinal plant Physalis divaricata (Solanaceae). Biodiversitas 18: 1430-1437. Physalis divaricata (Solanaceae) is one of the most prevalent weeds in summer crops. Putrescine Nmethyltransferase (PMT) is a key enzyme in the biosynthesis of nicotine, tropane alkaloids atropine, scopolamine, cocaine, and calystegines. The present study set to compare PMT gene expression during different growth stages of Ph. divaricata using RT-qPCR assay. RNA extraction was performed from root and leaf samples of a total number of 40 individuals Ph. divaricata at different growth stages (late vegetative and fruiting stages) collected from southwestern Iran. RT-qPCR of cDNA reversely synthesized from RNA was carried out using SYBR®Premix Ex TaqTM II kit. PMT gene expression levels were analyzed using ΔΔCT method. The results showed that expression level of PMT in late vegetative stage samples was significantly higher compared to fruiting stage samples. The expression level of PMT similarly changed in root and leaf samples. Direct visualization of alkaloids in different tissues using Wagner histochemical tests showed more concentrations of alkaloids in leaf idioblasts and root stele.

Tropane alkaloids are a wide group of substances that comprises more than 200 compounds occurring especially in the Solanaceae family. The main aim of this study is the development of a method for the analysis of the principal tropane alkaloids as atropine, scopolamine, anisodamine, tropane, tropine, littorine, homatropine, apoatropine, aposcopolamine, scopoline, tropinone, physoperuvine, pseudotropine and cuscohygrine in cereals and related matrices. For that, a simple solid-liquid extraction was optimized and a liquid chromatographic method coupled to a single stage Exactive-Orbitrap was developed. The method was validated obtaining recoveries in the range of 60-109% (except for some compounds in soy), precision values (expressed as relative standard deviation) lower than 20% and detection and quantification limits equal to or lower than 2 and 3μg/kg respectively. Finally, the method was applied to the analysis of different types of samples as buckwheat, linseed, soy and millet, obtaining positives for anisodamine, scopolamine, atropine, littorine and tropinone in a millet flour sample above the quantification limits, whereas atropine and scopolamine were detected in a buckwheat sample, below the quantification limit. Contaminated samples with Solanaceaes seeds (Datura Stramonium and Brugmansia Arborea) were also analysed, detecting concentrations up to 693μg/kg (scopolamine) for contaminated samples with Brugmansia seeds and 1847μg/kg (atropine) when samples were contaminated with Stramonium seeds.

Tetraploidy improves overexpression of h6h and scopolamine production of H. muticus, while in H. senecionis, pmt overexpression and elicitation can be used as effective methods for increasing tropane alkaloids. The effects of metabolic engineering in a polyploid context were studied by overexpression of h6h in the tetraploid hairy root cultures of H. muticus. Flow cytometry analysis indicated genetic stability in the majority of the clones, while only a few clones showed genetic instability. Among all the diploid and tetraploid clones, the highest level of h6h transgene expression and scopolamine accumulation was interestingly observed in the tetraploid clones of H. muticus. Therefore, metabolic engineering of the tropane biosynthetic pathway in polyploids is suggested as a potential system for increasing the production of tropane alkaloids. Transgenic hairy root cultures of Hyoscyamus senecionis were also established. While overexpression of pmt in H. senecionis was correlated with a sharp increase in hyoscyamine production, the h6h-overexpressing clones were not able to accumulate higher levels of scopolamine than the leaves of intact plants. Applying methyl jasmonate was followed by a sharp increase in the expression of pmt and a drop in the expression of tropinone reductase II (trII) which consequently resulted in the higher biosynthesis of hyoscyamine and total alkaloids in H. senecionis.

Aromatic amino acid aminotransferases (AAA-ATs) catalyze the reversible transamination reactions of proteinogenic and non-proteinogenic aromatic amino acids to corresponding keto acids and vice versa. The products of plant AAA-ATs serve as key precursors of many primary and secondary metabolites that are crucial for both plant and human metabolism and physiology. In most microbes, l-tyrosine (Tyr) and l-phenylalanine (Phe) aminotransferases (Tyr and Phe-ATs) catalyze the final steps of Phe and Tyr biosynthesis. On the other hand, plants use different pathways to synthesize Tyr and Phe via arogenate, in which prephenate-specific aminotransferases (PPA-ATs) catalyze the committed step in the plastids. Plant Tyr and Phe-ATs, unlike microbial counterparts, often prefer the reverse reactions and metabolize Tyr and Phe to their respective aromatic keto acids, which serve as precursors of various plant natural products (e.g. benzenoid volatiles, tocochromanols, plastoquinone, and tropane and benzylisoquinoline alkaloids). Unlike plastidic PPA-ATs, plant Tyr/Phe-ATs are localized outside of the plastids, have broad substrate specificity, and interlink Tyr and Phe metabolism. l-Tryptophan (Trp) aminotransferases (Trp-ATs) are involved in biosynthesis of the plant hormone auxin. Although significant advancement has been made on biochemical, molecular, and genetic characterizations of plant AAA-ATs, there are still many critical knowledge gaps, which are highlighted in the current review.

The objective of this study was to investigate the effects of 2,3,4-trimethoxy-N'-(8-metyl-8-azabicyclo[3.2.1.] octan-3-ylidene) benzohydrazide hydrochloride (LK-933), a compound of the tropane group with anxiolytic and antimigraine activity, on the levels of monoamines and their metabolites in different brain structures of outbred mice using HPLC–ED. Administration of LK-933 caused an increase in the composite characteristics that describe the rate of dopamine transformation into its metabolites dioxyphenylacetic acid and homovanillic acid in the frontal cortex by 114 and 181%, respectively. The levels of homovanillic acid in the same structure increased by 49%. These data implicate the dopaminergic pathway in the mechanism of LK-933 action.

Cocaine, a widely abused psychostimulant, inhibits the dopamine transporter (DAT) by trapping the protein in an outward-open conformation, whereas atypical DAT inhibitors such as benztropine have low abuse liability and prefer less outward-open conformations. Here, we use a spectrum of computational modeling and simulation approaches to obtain the underlying molecular mechanism in atomistic detail. Interestingly, our quantum mechanical calculations and molecular dynamics (MD) simulations suggest that a benztropine derivative JHW007 prefers a different stereoisomeric conformation of tropane in binding to DAT compared to that of a cocaine derivative, CFT. To further investigate the different inhibition mechanisms of DAT, we carried out MD simulations in combination with Markov state modeling analysis of wild-type and Y156F DAT in the absence of any ligand or the presence of CFT or JHW007. Our results indicate that the Y156F mutation and CFT shift the conformational equilibrium toward an outward-open conformation, whereas JHW007 prefers an inward-occluded conformation. Our findings reveal the mechanistic details of DAT inhibition by JHW007 at the atomistic level, which provide clues for rational design of atypical inhibitors.

BACKGROUND Tropane alkaloids are known to play a role in plant defence. By blocking acetylcholine receptors, they exert insecticidal and deterrent effects against herbivore insects. Carbamates are an important class of chemical insecticides that also inhibit acetyl cholinesterase. The objective of this work was to synthesise a series of tropane alkaloids bearing a carbamate group, and to evaluate their effects against the pest Ascia monuste. The effects of the most active compounds were evaluated on the A. monuste predator Solenopsis saevissima and on the pollinator Tetragonisca angustula. RESULTS The synthesis of carbamate-tropane alkaloids was accomplished in 4–5 steps from commercially available ketones. Results from bioassays showed that compounds 6a, 10a and 14a presented higher activities against second-instar larvae of A. monuste, with LD50 values of 1.01, 3.76 and 1.92 µg substance mg−1 insect, and TL50 values of 7.0, 15.0 and 5.0 h respectively. These compounds were also tested for their selectivity in favour of S. saevissima and T. angustula. Compound 6a, which showed the highest activity against A. monuste, also showed lower toxicity against S. saevissima. CONCLUSION Tropane alkaloid derivatives bearing a carbamate group show potential for the development of novel insecticides against A. monuste. © 2017 Society of Chemical Industry

Background Chronic obstructive pulmonary disease (COPD) is a heterogeneous disorder characterized by usually progressive development of airflow obstruction that is not fully reversible. While most patients will experience symptoms throughout the day or in the morning upon awakening, many patients do not experience their symptoms as constant but report variability in symptoms during the course of the day or over time. Symptom variability adversely affects patients' health status and increases the risk of COPD exacerbations. Methods We examined data from the literature on symptom variability and control in patients with COPD, with focus on the use of inhaled bronchodilator therapy with long-acting muscarinic antagonist agents (LAMA) plus long-acting β 2 -agonists (LABA); in particular twice-daily fixed-dose combination LAMA/LABA therapy with aclidinium/formoterol. Results Correct diagnosis and assessment of COPD requires comprehensive clinical and functional evaluation and consideration of individual needs to support the clinical decisions necessary for effective long-term management. Combining bronchodilators from different and complementary pharmacological classes with distinct mechanisms of action can increase the magnitude of bronchodilation as opposed to increasing the dose of a single bronchodilator. Conclusions The use of inhaled bronchodilator therapy with LAMA/LABA fixed-dose combinations in patients with stable COPD is supported by current evidence. This treatment approach provides robust effects on lung function and symptom control and may improve patients' adherence to treatment. Administration of the long-acting bronchodilators aclidinium and formoterol as twice daily fixed-dose aclidinium/formoterol 400/12 μg has the potential to control symptoms throughout the 24 h in patients with stable moderate-to-severe COPD.

Yeast has been engineered to convert simple sugars and amino acids into drugs that inhibit a neurotransmitter molecule. The work marks a step towards making the production of these drugs more reliable and sustainable. An engineered strain of yeast produces tropane alkaloids.

In this study, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a broad-spectrum monoclonal antibody for tropane alkaloids (TAs) was established for the rapid screening of atropine, scopolamine, homatropine, apoatropine, anisodamine, anisodine and L-hyoscyamine residues in pig urine, pork and cereal flour samples through a simple sample preparation procedure. The half inhibitory concentrations of atropine, homatropine, L-hyoscyamine, apoatropine, scopolamine, anisodamine and anisodine were 0.05, 0.07, 0.14, 0.14, 0.24, 5.30 and 10.15 ng mL−1, respectivelyThe detection and quantitative limits of this method for TAs in samples were 0.18–73.18 and 0.44–74.77 μg kg−1. The spiked recoveries ranged from 69.88% to 147.93%, and the coefficient of variations were less than 14%. Good correlation (R2 = 0.9929) between the results of the ic-ELISA and the high performance liquid chromatography-tandem mass spectrometry support the reliability of the developed ic-ELISA method.

Alkaloids are a large cluster of molecules found in Mother Nature all over the world. They are all secondary compounds and collection of miscellaneous elements and biomolecules, derived from amino acids or from transamination. This diverse chemical group is categorized, based on the amino acids that deliver their nitrogen atom and part of their skeleton. Alkaloids from a similar origin or having the same basic nucleus may have dissimilar biosynthetic pathways and different biological activity. They are derived from l -phenylalanine, l -tyrosine, anthranilic acid or acetate, l -histidine, l -ornithine, nicotinic acid, and l -lysine. Apart from other types of alkaloids, indole, tropane, and isoquinoline alkaloids are very important. People from all over the world are using them in their everyday life. Alkaloids can also have an animal origin, which may be endogenous or exogenous. This chapter highlights the phytochemistry of the main representatives among the diverse group of alkaloids. This chapter also focuses on their extraction, purification, fractionation, identification, and quantification procedures. A fundamental understanding of the biological activity of important alkaloids is also highlighted in this chapter. With the potential of revealing new compounds and diverse pharmacological properties, alkaloids still embrace a great potential for the future of drug discovery.

Higher plants produce divergent classes of specialized (so called secondary) metabolites, such as alkaloids, phenolics, terpenoids, and so on. Among them, alkaloids, which contain nitrogen atom often in heterocyclic ring, are most biologically active and used as poisons and/or also pharmaceuticals. Whereas natural resources are commonly used for the preparation for these chemicals, shortage of natural resources in quantity and quality need more advanced technology for the production. Recent progresses in biochemistry, molecular biology and recombinant technology provide new way for the production of specialized metabolites without chemical synthesis. Metabolic engineering in target plants and reconstruction of biosynthetic pathway in heterologous hosts, especially microbes using synthetic biology, offers great potentials to improve the productivity and quality of these metabolites. In this report, recent progresses in the isolation and identification of molecular tools for metabolic engineering and synthetic biology are critically summarized and also some achievement of alkaloid engineering in benzylisoquinoline alkaloids (BIAs) such as berberine, morphine, noscapine and sanguinarine, monoterpenoid indole alkaloids (MIAs) such as vinca alkaloids, nicotine and tropane alkaloids, steroidal glycoalkaloids such as solanine and so on are summerized.

Egyptian henbane (Hyoscyamus muticus L.) plants are rich sources of alkaloids used in pharmaceutical products. Recently, rising efforts have been devoted to reducing mineral fertilizer supply, production cost, and environmental pollution via decreasing the doses of nitrogenous fertilizers and adopting biofertilizer farming systems. Two field experiments were conducted to examine the potential role of N fixing bacteria Azotobacter spp. and Azospirillum spp. on the growth, mineral status, tropane alkaloids, leaf anatomy, and seed yield of Egyptian henbane grown with different levels of mineral nitrogen fertilizer, i.e., 25%, 50%, and 100% of the recommended dose, equal to 30, 60, and 120 kg N ha&minus, 1. N fertilizer improved growth, mineral elements, tropane alkaloids, seed yield, and yield components of Egyptian henbane, which showed a gradually rising trend as the rate of N fertilizer increased. High doses of N fertilizer presumably elicited favorable changes in the anatomical structure of Egyptian henbane leaves. The application of 50% N dose plus N fixing bacteria affected Egyptian henbane trials similarly to 100% of recommended N dose. In conclusion, the N fixing bacteria proved to be a sustainable tool for a two-fold reduction in the recommended dose of mineral N fertilizer and the sustainable management of Egyptian henbane nutrition.

The medicinal use of tropane alkaloids is well established and has a long history of application, according to the anticholinergic activity of these compounds. Among them, hyoscyamine, anisodamine (6β-hydroxyhyoscyamine), and scopolamine are the most important ones from a therapeutical point of view. Although the chemical synthesis of these alkaloids is possible, its complexity, costs, and low quality of the compounds obtained make it a nonviable strategy for a commercial production. For the reason mentioned above, tropane alkaloids are still extracted from plants grown in greenhouses belonging to species of Solanaceae family including Hyoscyamus niger L., Anisodus tanguticus, Scopolia tangutica Maxim, Atropa belladonna, and several Datura species. In the last years, considerable effort has been devoted to develop cost-effective strategies for their production. Tropane alkaloid production by in vitro culture techniques, such as hairy roots, is an interesting alternative since it guarantees a stable and continuous production throughout the year, independent of the presence of pathogens as well as the environmental conditions. This chapter will focus and discuss the establishment of hairy root cultures for the production of tropane alkaloids in order to replace the isolation from the natural source. In addition, recent progress achieved in the application of elicitation strategies for improving their release to the culture medium will be analyzed.