Spirolactone Ring Research Articles

A new phenanthrene with a spirolactone ring from Dendrobium ochreatum

Dendroochreatene (1), a new phenanthrene derivative with a spirolactone ring, was isolated from the whole Dendrobium ochreatum plant together with 11 known compounds (2–12). The structure of the new compound was elucidated spectroscopically and phenolic compounds were firstly reported from D. ochreatum. Moscatilin (4), major compound isolated from D. ochreatum, was found to be cytotoxic toward H460 lung-cancer cells, with an IC50 value of 147.3 ± 0.9 µM, while loddigesiinol C (7), C-α-methoxy derivative was inactive.

Structure-diversified terpenoids from Salvia prattii and their protective activity against alcoholic liver diseases

Eleven previously unreported compounds (1–11), including five diterpenoids (1–5) and six sesquiterpenoids (6–11), together with two known diterpenoids (12–13), have been isolated from the roots of Salvia prattii. Their structures were comprehensively elucidated through spectroscopic methods, and their configurations were established using computational 13C nuclear magnetic resonance and electronic circular dichroism. Compound 1 was found to be an abietane-type diterpenoid with a novel rearrangement generated from the cleavage of the C-4/5 chemical bond, 20-methyl shift, and the rearrangement of the C-10 side chain. Compounds 2–3 were the third and fourth examples of arrangement seco-norabietanes with a spiro-lactone ring. We evaluated all compounds for their protective effects against alcoholic liver diseases (ALD). Compound 2 exhibited potential protective activity and hence can be used as a novel anti-ALD candidate.

Synthesis of two novel fluorescein appended dipyromethanes (DPMs): Naked-eye chemosensors for fluoride, acetate and phosphate anions

Two novel fluorescein based dipyromethane probes DPM1 and DPM2 were constructed from a simple and easily accessible fluorescein dye, and were fully characterized by means of standard spectroscopic techniques. Studies of the DPM1 and DPM2, investigated using UV–vis spectroscopy with different anions in CH3CN/DMSO, displayed selective colorimetric sensing towards fluoride, acetate and phosphate anions. Moreover, the fluorescence analysis was also accomplished with DPM1/DPM2, revealing the enhancement in the emission spectra of the fluoride, acetate and phosphate anions. From the studies, we noticed that the DPM1/DPM2 having spirolactone ring is non-fluorescent, which upon addition of the fluoride, phosphate and acetate, underwent ring-opening, resulting in the generation of strong fluorescence. The solid state images divulges that DPM1 shows green fluorescence and DPM2 displayed dark green fluorescence upon irradiating with UV-light of wavelength 254 nm. We infer that such types of naked eye sensors might reveal the potential applications in the selective and sensitive detection of fluoride/acetate/phosphate, and/or other biologically significant anions in nearby future.

Secondary Metabolites from Dendrobium nobile and Their Activities Induce Metabolites Apoptosis in OSC-19 Cells.

To identify potential drug candidates, secondary metabolites of Dendrobium nobile were performed. As a result, two previously undescribed phenanthrene derivatives with a spirolactone ring (1 and 2), along with four known compounds, N-trans-cinnamoyltyramine (3), N-trans-p-coumaroyltyramine (4), N-trans-feruloyltyramine (5), and moscatilin (6), were isolated from Dendrobium nobile. The structures of the undescribed compounds were elucidated using NMR spectroscopy, electronic circular dichroism (ECD) calculations, and extensive spectroscopic data analysis. The cytotoxic effects of compounds on human tongue squamous cells OSC-19 were determined using MTT at concentrations of 2.5 μM, 5 μM, 10 μM, and 20 μM. Compound 6 exhibited potent inhibitory activity against OSC-19 cells with an IC50 of 1.32 μM. Migration assays and western blot assays demonstrated that compound 6 effectively inhibited migration by down-regulating MMP2 and MMP9 at concentrations of 0.5 μM and 1 μM. To investigate its effect on apoptosis, we performed AO/PI staining, flow cytometry, and WB experiments. The results showed that increasing concentrations led to increased red fluorescence, decreased green fluorescence, increased apoptosis rate, decreased expression of bcl-2, caspase 3, caspase 9, and parp proteins, and increased bax expression. Furthermore, the phosphorylation of JNK and P38 was activated, suggesting that compound 6 may induce apoptosis via the MAPK pathway.

Anti-inflammatory maistemonine-class alkaloids of Stemona japonica

Three hitherto undescribed Stemona alkaloids, named stemajapines A–C (1–3), along with six known alkaloids (4–9), were isolated and identified from the roots of Stemona japonica (Blume) Miq. (Stemonaceae). Their structures were established by the analysis of the mass data, NMR spectra, and computational chemistry. Stemjapines A and B were degraded maistemonines without spiro-lactone ring and skeletal methyl from maistemonine. Concurrence of alkaloids 1 and 2 revealed an undescribed way to form diverse Stemona alkaloids. Bioassay results disclosed the anti-inflammatory natural constituents stemjapines A and C with IC50 values of 19.7 and 13.8 µM, respectively, compared to positive control dexamethasone with 11.7 µM. The findings may point out a new direction of Stemona alkaloids inaddition to its traditional antitussive and insecticide activities.

Mercury ion-selective fluorescent probe based on indazole fused rhodamine and cell imaging application

Developing selective and sensitive probes for mercury ion (Hg2+) is of great significance because Hg2+ exhibits high toxicity and poses a potential threat to public health and the environment. Herein, a newly designed indazole fused rhodamine dye InRh has been prepared, which shows stable fluorescent signals in water at a wide pH range of 1.0–10.0. Based on the dye InRh, a novel fluorescent probe InRhH for Hg2+ is further developed by modifying the spirolactone ring of InRh to form a closed spirolactam ring. The probe shows selective fluorescence “off–on” signal changes to Hg2+ with a visible color change and response time of 40 min in H2O/CH3CN (pH = 6.0, v/v = 4/1), and the detection limit for Hg2+ is measured to be 0.33 nM. The recognition mechanism for Hg2+ can be ascribed to the binding interactions between Hg2+ and probe in a 2: 1 coordination mode, which promotes the occurrence of spirolactam ring opening and further hydrolysis. In addition, the InRhH can be applied for test strip experiments and tracing 10−6 M level Hg2+ with visible color change. Furthermore, the suitable cytotoxicity allows the probe to apply for sensing Hg2+ in living HeLa cells by fluorescence imaging.

A dual-response fluoran-phenothiazine hybrid fluorescent probe for selective sensing of Fe3+ and ClO− and cell imaging application

Bifunctional fluorescent probes with dual-emission response attract extensive attention. A novel fluorescent probe FP, a hybrid of fluoran and phenothiazine, has been designed and synthesized for selective sensing of Fe3+ and ClO− with dual-emission changes, which involes mechanisms of Fe3+-promoted spirolactone ring opening and ClO−-induced oxidation of phenothiazine moiety, respectively. In addition, the detection limits for Fe3+ and ClO− were estimated to be 49.1 and 35.9 nM, respectively. Significantly, FP can be employed as an tracer for the detection of Fe3+ ions within living HeLa cells by fluorescence imaging.

A Deep-Red to Near Infrared (NIR) Fluorescent Probe Based on a Sulfur-Modified Rhodamine Derivative with Two Spirolactone Rings.

We report the synthesis, characterization, and photophysical and ion-binding properties of deep-red to near-infrared (NIR) fluorescent rhodamine derivatives, bearing two spirolactone rings and substitution of the oxygen atoms in the xanthene ring with sulfur atoms (1-S). The diastereoisomeric cis- and trans-forms of the rhodamine derivative were separated and the cis-form (cis-1-S) was structurally characterized by X-ray crystallography. Upon treatment with Hg2+ ion, cis-1-S was converted into the dual spirolactone ring-opened species, resulting in significant color change and fluorescence enhancement. Substitution of the oxygen atoms with sulfur and extended π-conjugation across the fused six-membered rings upon the two rings-opening processes in the presence of Hg2+ ion led to a significant red-shift of absorption (623 nm) and fluorescence (706 nm) peaks, compared to the ordinary rhodamine. Furthermore, the intracellular Hg2+ -sensing properties of the cis-1-S have been studied by confocal microscopy.

Rhodol-conjugated polymersome sensor for visual and highly-sensitive detection of hydrazine in aqueous media

Hydrazine is a hazardous environmental pollutant, which contaminates land, air and water posturing a severe risk to human health. For the first-hand estimation, a qualitative approach (colorimetric) for recognition of hydrazine could suffice. However, for accurate measurement, under the threshold limit value (TLV), a quantitative technique is desired. We report the polymersome-based sensor for visual detection and quantification of hydrazine in water. The rhodol-functionalized amphiphilic hyperbranched multiarm copolymer (HSP-RDL) was self-assembled into vesicles. The HSP-RDL vesicle probe exhibited high sensitivity and selectivity for hydrazine recognition in presence of various competitive species such as cations, anions, and neutral species. The fast responsive pink color change from colorless could be visualized with naked eye due to spirolactone ring opening by hydrazinolysis triggered strong fluorescence emission. The vesicle probe could detect hydrazine in water with a limit of detection (LOD) value of 2 nM (0.0652 ppb), which is lower than TLV (10 ppb) given by USEPA (United States Environmental Protection Agency). Furthermore, the vesicle probe could quantify hydrazine (recovery ≥ 99 %) in a wastewater sample collected from Huangpu river. The membrane-permeable characteristics of HSP-RDL led hydrazine detection in live cells through confocal fluorescence microscopy.

Novel π-extended hybrid xanthene dyes with two spirolactone rings for optoelectronic and biological applications

The design, synthesis, and photophysical properties of organic fluorophores have attracted considerable research interest due to the utility of these compounds for various optoelectronic, analytical, and biological applications. In this study, we synthesized two novel π-extended red-emitting hybrid xanthene dyes, each of which has two spirolactone rings and combines a seminaphthofluorescein moiety and a seminaphthorhodafluor moiety in a single molecule. The photophysical properties of the dyes in methanol in the presence of acid, base, and metal cations were investigated. Mono-ring-opened seminaphthofluorescein and seminaphthorhodafluor forms of the dyes could be obtained by the addition of OH- or H+, respectively. Owing to the changes in the absorbance spectra of the mono-ring-opened forms induced by addition of H+ and OH-, the dyes could perform simultaneously the functions of an XOR gate and an INHIBIT gate, with the absorbances at 510 and 560 nm as outputs, respectively, and could act as half-subtractors with H+ and OH- as inputs. Furthermore, stepwise ring-opening could be induced by Hg2+ ions in methanol. In water, the dyes existed in double-ring-opened forms that emitted deep-red fluorescence and were mitochondria-targetable, suggesting that these chromophores might be useful as fluorescence tracers in biological applications. Because the absorption and fluorescence properties of these fluorophores can be regulated via their two spirolactone rings, we expect that these compounds will find utility in various optoelectronic, analytical, and biological applications.

Discovery of an Extended Austinoid Biosynthetic Pathway in Aspergillus calidoustus.

Filamentous fungi produce a wide range of natural products that are commonly used in various industrial contexts (e.g., pharmaceuticals and insecticides). Meroterpenoids are natural products of interest because of their various biological activities. Among the meroterpenoids, there is a group of insecticidal compounds known as the austinoids. These compounds have also been studied because of their intriguing spiro-lactone ring formation along with various modifications. Here, we present an extension of the original austinol/dehydroaustinol biosynthesis pathway from Aspergillus nidulans in the recently identified filamentous fungus Aspergillus calidoustus. Besides the discovery and elucidation of further derivatives, genome mining led to the discovery of new putative biosynthetic genes. The genes involved in the biosynthesis of later austinoid products were characterized, and among them was a second polyketide synthase gene in the A. calidoustus cluster that was unusual because it was a noninterative polyketide synthase producing a diketide. This diketide product was then loaded onto the austinoid backbone, resulting in a new insecticidal derivative, calidodehydroaustin.

Fused-Fluoran Leuco Dyes with Large Color-Change Derived from Two-Step Equilibrium: iso-Aminobenzopyranoxanthenes.

Fluorans are popular leuco dyes that are used in various applications, such as carbonless-copy papers and thermal papers. Here, we describe unique leuco dyes in which two fluoran units are fused into a C2h structure (iso-aminobenzopyranoxanthenes; iso-ABPXs). iso-ABPXs exhibited a large two-step color change (colorless/pink and pink/blue-green) due to opening-closing of two spirolactone rings. The two-step equilibrium among the colorless, pink, and blue-green forms could be well controlled by adjusting acid concentration, solvent, and/or temperature.

A Molecular Chameleon with Fluorescein and Rhodamine Spectroscopic Behaviors.

A new class of fluorescein/rhodamine hybrids with two spirolactone rings was reported to exhibit dual-output fluorescent behaviors independently. Isolation and characterization for two diastereomers, trans-RhOH and cis-RhOH, have been made and their X-ray crystal structures determined. In a basic environment, the spirolactone ring on the hydroxyl side will be opened to give a fluorescein-like optical output with the lowest absorptions at 485 and 530 nm emission. On the other hand, a rhodamine-like optical output, i.e., 528 nm absorption and 575 nm emission, will be switched on by a H(+) or a Hg(2+) ion, attributed to the spirolactone ring opening on the amino side. In a methanol-buffer system with different pH values, the corresponding pKa values for the hydroxyl and amino groups were determined as 5.7 and 2.3, respectively. Selective Hg(2+)-sensing properties have also been discussed, and log Ks values of about 3.60 and 3.73 were determined. Confocal microscopic images of Caenorhabditis elegans incubated with RhOH were found to show enhanced fluorescent intensity with a Hg(2+) ion, demonstrating the potential application of RhOH for in vivo biological imaging.

A rhodamine B-based fluorescent sensor toward highly selective mercury (II) ions detection

This work presented the design, syntheses and photophysical properties of a rhodamine B-based fluorescence probe, which exhibited a sensitive and selective recognition towards mercury (II). The chemosensor RA (Rhodamine- amide- derivative) contained a 5-aminoisophthalic acid diethyl ester and a rhodamine group, and the property of spirolactone of this chemosensor RA was detected by X-ray crystal structure analyses. Chemosensor RA afforded turn-on fluorescence enhancement and displayed high brightness for Hg2+, which leaded to the opening of the spirolactone ring and consequently caused the appearance of strong absorption at visible range, moreover, the obvious and characteristic color changed from colorless to pink was observed. We envisioned that the chemosensor RA exhibited a considerable specificity with two mercury (II) ions which was attributed to the open of spirolactone over other interference metal ions.

Analysis of Chemical Equilibrium of Silicon-Substituted Fluorescein and Its Application to Develop a Scaffold for Red Fluorescent Probes

Fluorescein is a representative green fluorophore that has been widely used as a scaffold of practically useful green fluorescent probes. Here, we report synthesis and characterization of a silicon-substituted fluorescein, i.e., 2-COOH TokyoMagenta (2-COOH TM), which is a fluorescein analogue in which the O atom at the 10' position of the xanthene moiety of fluorescein is replaced with a Si atom. This fluorescein analogue forms a spirolactone ring via intramolecular nucleophilic attack of the carboxylic group in a pH-dependent manner. Consequently, 2-COOH TM exhibits characteristic large pH-dependent absorption and fluorescence spectral changes: (1) 2-COOH TM is colorless at acidic pH, whereas fluorescein retains observable absorption and fluorescence even at acidic pH, and the absorption maximum is also shifted; (2) the absorption spectral change occurs above pH 7.0 for 2-COOH TM and below pH 7.0 for fluorescein; (3) 2-COOH TM shows a much sharper pH response than fluorescein because of its pKa inversion, i.e., pKa1 > pKa2. These features are also different from those of a compound without the carboxylic group, 2-Me TokyoMagenta (2-Me TM). Analysis of the chemical equilibrium between pH 3.0 and 11.0 disclosed that 2-COOH TM favors the colorless and nonfluorescent lactone form, compared with fluorescein. Substitution of Cl atoms at the 4' and 5' positions of the xanthene moiety of 2-COOH TM to obtain 2-COOH DCTM shifted the equilibrium so that the new derivative exists predominantly in the strongly fluorescent open form at physiological pH (pH 7.4). To demonstrate the practical utility of 2-COOH DCTM as a novel scaffold for red fluorescent probes, we employed it to develop a probe for β-galactosidase.

Photophysical and Laser Properties of Cassettes based on a BODIPY and Rhodamine Pair

This work deals with the synthesis and the photophysical and laser properties of new BODIPY-rhodamine cassettes. These dyads differ in their rigid and conjugated spacer group (phenyl or acetylenephenyl) and in their linking positions (meta or para). The photophysical properties of these cassettes are controlled by the formation/opening of the spirolactone ring, which, in turn, switches off/on an energy-transfer process between the chromophores. Herein, we thoroughly describe the influence of the attached spacer group, as well as the distance and orientation between the donor-acceptor pair, on the excitation energy transfer. The observed fast dynamics and efficiency suggest that the process mainly takes place "through-bond", although the "through-space" mechanism also contributes to the whole process. As a result, efficient laser emission from the rhodamine is achieved upon excitation of the BODIPY, in particular for the cassette that contains an acetylenephenyl spacer group in a para disposition.

Spiro-Ring Formation is Catalyzed by a Multifunctional Dioxygenase in Austinol Biosynthesis

Austinol, a fungal meroterpenoid derived from 3,5-dimethylorsellinic acid, has a unique chemical structure with a remarkable spiro-lactone ring system. Despite the recent identification of its biosynthetic gene cluster and targeted gene-deletion experiments, the process for the conversion of protoaustinoid A (2), the first tetracyclic biosynthetic intermediate, to the spiro-lactone preaustinoid A3 (7) has remained enigmatic. Here we report the mechanistic details of the enzyme-catalyzed, stereospecific spiro-lactone ring-forming reaction, which is catalyzed by a non-heme iron-dependent dioxygenase, AusE, along with two flavin monooxygenases, the 5'-hydroxylase AusB and the Baeyer-Villiger monooxygenase AusC. Remarkably, AusE is a multifunctional dioxygenase that is responsible for the iterative oxidation steps, including the oxidative spiro-ring-forming reaction, to produce the austinol scaffold.

Preparation and antitrypanosomal activity of secochiliolide acid derivatives

Secochiliolide acid (1) isolated from the Patagonian shrub Nardophyllum bryoides, was used as a scaffold for the preparation of a series of nine derivatives. Compound 1 and its derivatives were tested against Trypanosoma cruzi epimastigotes grown in liquid media. It was first observed that secochiliolide acid (1) inhibited the proliferation of the parasites, with an IC50 of 2μg/mL. Six of the synthesized derivatives were also active with IC50’s between 2 and 7μg/mL which are comparable to that of the commercial drug benznidazole (2.5μg/mL). These results indicate that the carboxyl group is not essential for the bioactivity of 1, while the presence of the tetrasubstituted exocyclic double bond seems to be important. Moreover, the presence of the furan and spirolactone rings is not essential for the bioactivity per se, but is important in combination with other structural fragments present in the molecule.

Design, synthesis, and structure–activity relationships of novel spiro-piperidines as acetyl-CoA carboxylase inhibitors

Spiro-lactone (S)-1 is a potent acetyl-CoA carboxylase (ACC) inhibitor and was found to be metabolically liable in human hepatic microsomes. To remove one of the risk factors in human study by improving the metabolic stability, we focused on modifying the spiro-lactone ring and the benzothiophene portion of the molecule. Spiro-imide derivative 8c containing a 6-methylthieno[2,3-b]pyridine core exhibited potent ACC inhibitory activity and favorable pharmacokinetic profiles in rats.

An efficient total synthesis of Stemonamine

A short and efficient approach to the Stemona alkaloids has been developed. The representative members, Stemonamine, has been synthesized from the known key intermediate 1 in 7 steps and 47.3% overall yield featured by a novel ketone-ester condensation to forge the spirolactone ring.