One-Pot Fluorescent Labeling Protocol for Complex Hydroxylated Bioactive Natural Products

Haldar, Saikat; Kumar, Santosh; Kolet, Swati P.; Patil, Harshal S.; Kumar, Dhiraj; Kundu, Gopal C.; Thulasiram, Hirekodathakallu V.

doi:10.1021/jo401559t

Cited by 7 publications

(9 citation statements)

References 39 publications

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“…Instead, elimination of the C-14 acetoxyl group had occurred. It was later found that Yamaguchi esterification protocol (2,4,6-trichlorobenzoyl chloride/TEA/DMAP/THF) [ 21 ] could successfully give ANDRO-NBD in moderate yield (54%).…”

Section: Resultsmentioning

confidence: 99%

Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

Hsu¹,

Hsu

Liu

et al. 2016

PLoS ONE

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Andrographolide (ANDRO) is a lactone diterpenoid compound present in the medicinal plant Andrographis paniculata which is clinically applied for multiple human diseases in Asia and Europe. The pharmacological activities of andrographolide have been widely demonstrated, including anti-inflammation, anti-cancer and hepatoprotection. However, the pharmacological mechanism of andrographolide remains unclear. Therefore, further characterization on the kinetics and molecular targets of andrographolide is essential. In this study, we described the synthesis and characterization of a novel fluorescent andrographolide derivative (ANDRO-NBD). ANDRO-NBD exhibited a comparable anti-cancer spectrum to andrographolide: ANDRO-NBD was cytotoxic to various types of cancer cells and suppressed the migration activity of melanoma cells; ANDRO-NBD treatment induced the cleavage of heat shock protein 90 (Hsp90) and the downregulation of its client oncoproteins, v-Src and Bcr-abl. Notably, ANDRO-NBD showed superior inhibitory effects to andrographolide in all anticancer assays we have performed. In addition, ANDRO-NBD was further used as a fluorescent probe to investigate the uptake kinetics, cellular distribution and molecular targets of andrographolide. Our data revealed that ANDRO-NBD entered cells rapidly and its fluorescent signal could be detected in nucleus, cytoplasm, mitochondria, and lysosome. Moreover, we demonstrated that ANDRO-NBD was covalently bound to several putative target proteins of andrographolide, including NF-κB and hnRNPK. In summary, we developed a fluorescent andrographolide probe with comparable bioactivity to andrographolide, which serves as a powerful tool to explore the pharmacological mechanism of andrographolide.

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Section: Resultsmentioning

confidence: 99%

Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

Hsu¹,

Hsu

Liu

et al. 2016

PLoS ONE

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“…All other chemicals of analytical grade were procured locally, in India. Extraction of neem limonoids AzaA from the neem seed kernel was performed as described by Alam et al 7-Nitrobenzo-2-oxa-1,3-diazole (NBD)-based labeled AzaA was used for gel electrophoresis experiments …”

Section: Materials and Methodsmentioning

confidence: 99%

“…Extraction of neem limonoids AzaA from the neem seed kernel was performed as described by Alam et al 39 7-Nitrobenzo-2-oxa-1,3-diazole (NBD)-based labeled AzaA was used for gel electrophoresis experiments. 40 Insect Feeding and Collection of Tissues. Actively feeding H. armigera larvae collected from fields were transferred and maintained on artificial diet (AD).…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Azadirachtin-A from Azadirachta indica Impacts Multiple Biological Targets in Cotton Bollworm Helicoverpa armigera

et al. 2019

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Azadirachtin-A (AzaA) from the Indian neem tree ( Azadirachta indica ) has insecticidal properties; however, its molecular mechanism remains elusive. The “targeted and nontargeted proteomic profiling”, metabolomics, matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) imaging, gene expression, and in silico analysis provided clues about its action on Helicoverpa armigera . Fourth instar H. armigera larvae fed on AzaA-based diet (AzaD) suffered from significant mortality, growth retardation, reduced larval mass, complications in molting, and prolonged development. Furthermore, death of AzaD-fed larvae was observed with various phenotypes like bursting, blackening, and half-molting. Liquid chromatography–mass spectrometry (LC–MS) data showed limited catabolic processing of ingested AzaA and dramatic alternations of primary metabolism in H. armigera . MALDI-TOF imaging indicated the presence of AzaA in midgut of H. armigera . In the gut, out of 79 proteins identified, 34 were upregulated, which were related to digestion, immunity, energy production, and apoptosis mechanism. On the other hand, 45 proteins were downregulated, including those from carbohydrate metabolism, lipid metabolism, and energy transfer. In the hemolymph, 21 upregulated proteins were reported to be involved in immunity, RNA processing, and mRNA-directed protein synthesis, while 7 downregulated proteins were implicated in energy transfer, hydrolysis, lipid metabolism, defense mechanisms, and amino acid storage-related functions. Subsequently, six target proteins were identified using labeled AzaA that interacted with whole insect proteins. In silico analysis suggests that AzaA could be efficiently accommodated in the hydrophobic pocket of juvenile hormone esterase and showed strong interaction with active site residues, indicating plausible targets of AzaA in H. armigera . Quantitative polymerase chain reaction analysis suggested differential gene expression patterns and partly corroborated the proteomic results. Overall, data suggest that AzaA generally targets more than one protein in H. armigera and hence could be a potent biopesticide.

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“…On the other hand, though the thiolysis of NBD probes in H 2 S detection has been successful, [31][32][33][34][35][36][37][38][39][40][41][42] the rising issue whether NBD-based dyes are stable in bioimaging should be addressed, because endogenous H 2 S exists in vivo and NBDbased dyes were widely used in bioimaging. [44][45][46][47][48] Herein, we prepared a series of NBD amines and studied their thiolysis reactions in buffer solution. The reaction rates of NBD amines towards H 2 S varied from 43.2 to 0.05 M −1 s −1 or smaller values dependent on the chemical structures.…”

Section: Introductionmentioning

confidence: 99%

Investigation of thiolysis of NBD amines for the development of H₂S probes and evaluating the stability of NBD dyes

Song

et al. 2016

Org. Biomol. Chem.

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In order to evaluate the thiolysis of NBD (7-nitro-1,2,3-benzoxadiazole) amines for development of HS probes, herein we investigated the reactivity and selectivity of a series of NBD amines for the first time. The piperazinyl- and piperidyl-based NBD probes could react efficiently with micromolar HS in buffer (pH 7.4), while such NBD(S) (nitrobenzothiadiazole) derivatives showed much slow thiolysis even in the presence of millimolar HS. Low reactivity was also observed for thiolysis of these ethylamino-, ethanolamino- and anilino-based NBD probes. Therefore, almost all NBD amines used in bioimaging should be stable, in consideration of the presence of only micromolar endogenous HS in vivo. Moreover, the piperazinyl-NBD derivatives could be efficient in the development of fluorescent HS probes and for directly visualizing HS by paper-based detection.

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One-Pot Fluorescent Labeling Protocol for Complex Hydroxylated Bioactive Natural Products

Cited by 7 publications

References 39 publications

Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

Azadirachtin-A from Azadirachta indica Impacts Multiple Biological Targets in Cotton Bollworm Helicoverpa armigera

Investigation of thiolysis of NBD amines for the development of H₂S probes and evaluating the stability of NBD dyes

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One-Pot Fluorescent Labeling Protocol for Complex Hydroxylated Bioactive Natural Products

Cited by 7 publications

References 39 publications

Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

Development of a Bifunctional Andrographolide-Based Chemical Probe for Pharmacological Study

Azadirachtin-A from Azadirachta indica Impacts Multiple Biological Targets in Cotton Bollworm Helicoverpa armigera

Investigation of thiolysis of NBD amines for the development of H2S probes and evaluating the stability of NBD dyes

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Investigation of thiolysis of NBD amines for the development of H₂S probes and evaluating the stability of NBD dyes