Search Results (29)

Ginkgo biloba seeds are a rich source of flavonoids and the unique terpene lactones—ginkgolides and bilobalide, known for their neuroprotective and cognitive-improving effects. However, unlike the widely used leaves, the seeds contain substantial levels of ginkgolic acid and ginkgotoxin (4′-O-methylpyridoxine), an antivitamin B₆ compound. At high concentrations, ginkgotoxin exhibits neurotoxicity, potentially inducing seizures, respiratory distress, and loss of consciousness, thus limiting the safe application of Ginkgo seed-derived products. This study aimed to develop a simple yet effective extraction protocol that reduces ginkgotoxin levels in Ginkgo biloba seed extracts while preserving their beneficial phytochemicals. A multistep liquid–liquid extraction approach employing sequential polar and non-polar solvents was implemented. Following each extraction stage, fractions were analyzed using ultra-high-performance liquid chromatography coupled with mass spectrometry (UHPLC–MS). The concentrations of flavonoids, ginkgolides, bilobalide, ginkgolic acid, and ginkgotoxin were quantified to evaluate detoxification efficiency and phytochemical retention. Compared with conventional single-step extraction using 70% methanol, this multistep protocol markedly reduced ginkgotoxin and ginkgolic acid to near-undetectable levels, while preserving detectable concentrations of major flavonoids and terpene trilactones. The findings demonstrate that multistep extraction represents a promising and practical strategy for minimizing ginkgotoxin in Ginkgo biloba seed extracts without compromising their beneficial phytochemical composition. This approach provides a sound basis for developing safer, functionally active Ginkgo-based products. Full article

Ginkgolic acids (GAs), the principal toxic constituents in Ginkgo biloba, pose health risks including cytotoxicity, allergenicity, and pro-inflammatory effects, limiting the application of Ginkgo resources in the food and health product industries. Developing efficient and environmentally friendly removal methods is essential. The endophytic fungus Fusarium sp. DLT-118, isolated from Ginkgo biloba, degraded 96.47% of GAs in Ginkgo biloba leaf extract (GE) at an initial concentration of 1 mg/mL within 7 days at 28 °C, while concurrently enhancing the antioxidant activity of GE, as indicated by a reduction in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging IC₅₀ from 755.7 μg/mL to 544.6 μg/mL. Morphological and oxidative stress analyses showed critical cellular adaptations and stress responses under degradation conditions. Integrated multi-omics analysis indicated that GE stress induced the remodeling of fungal amino acid, lipid, and energy metabolism, as well as the adjustment of membrane and transport functions, to facilitate GAs detoxification. Cytotoxicity assays indicated no significant cytotoxicity of the degradation products towards human normal lung epithelial cells (Beas-2B) and gastric mucosal epithelial cells (GES-1). These findings highlight Fusarium sp. DLT-118 as a promising agent for the efficient removal of GAs, offering a potential strategy for the production of GA-reduced Ginkgo-based food and health products. Full article

Ginkgolic acid (GA) exhibits various biological activities, but its role in vascular restenosis remains unreported. GA (13:0) is a relatively abundant natural congener. This study aims to investigate and clarify the effects and mechanisms of GA (13:0) on vascular smooth muscle cell (VSMC) proliferation and migration in vitro, as well as on balloon injury-induced vascular restenosis in rats. The results showed that GA (13:0) significantly inhibited VSMC proliferation, migration, and intimal thickening both in vitro and in vivo. Moreover, GA (13:0) reduced the expression of cyclin D1, cyclin E1, CDK2, and CDK4, as well as cyclin D1-CDK4 and cyclin E1-CDK2 binding, leading to G0/G1 arrest. Additionally, GA (13:0) suppressed vimentin expression and actin cytoskeleton polymerization and altered F-actin morphology. Comparative proteomics identified tectonic family member 1 (TCTN1) as a potential molecular target of GA (13:0). GA (13:0) reduced TCTN1 expression both in vitro and in vivo. Crucially, TCTN1 overexpression notably reversed the inhibitory effects of GA (13:0) on VSMC proliferation, migration, intimal thickening, expression and binding of cell cycle-related proteins, and vimentin expression. Concurrently, TCTN1 overexpression also reversed GA (13:0)-induced F-actin depolymerization and rearrangement and G0/G1 arrest. GA (13:0) significantly inhibited TCTN1 co-localization with vimentin and actin in vitro and in vivo. Furthermore, we found that CCCTC binding factor (CTCF) binds to the 162–176 site of the TCTN1 promoter to regulate TCTN1 transcription, and CTCF knockout significantly down-regulated TCTN1 protein levels. This study reveals that GA (13:0) inhibits TCTN1 transcription and expression, hindering G1/S transition, vimentin expression, and F-actin rearrangement, thereby suppressing vascular restenosis. Full article

This study established an efficient in vitro regeneration system using stem nodes from root collar suckers as explants. Subsequently, regenerated shoots were used to establish an in vitro medicinal production protocol that achieved ginkgolic acid production. The self-developed Ginkgo biloba medium (GBM), first reported in this study, was pivotal to system establishment. The plantlet propagation system showed that the bases of stem nodes dipped in GBM with 2 mg·L⁻¹ 6-benzyladenine (BA) and 0.2 mg·L⁻¹ 1-naphthaleneacetic acid (NAA) achieved near-complete axillary bud induction (99.56%). Adventitious shoot induction reached 82.22% (3.5 shoots/explant) using GBM with 0.2 mg·L⁻¹ BA, 0.02 mg·L⁻¹ kinetin (Kin) and 0.2 g·L⁻¹ proline (Pro). Maximum adventitious shoot elongation (92.22%, average 3.35 cm) was observed on GBM containing 0.1 mg·L⁻¹ zeatin (ZT) and 0.01 mg·L⁻¹ BA. After 3-week preculture with 15 mg·L⁻¹ phloroglucinol (PG), treatment with 0.6 mg·L⁻¹ indole-3-butyric acid (IBA) and 0.2% activated carbon (AC) yielded 96.67% rooting (6.19 roots/explant) and 85% acclimatization survival. For medicinal resource production, bud cluster induction at 94.44% (20.89 buds/explant) on GBM with 1 mg·L⁻¹ BA, 0.03 mg·L⁻¹ Kin, and 0.2 g·L⁻¹ Pro. Leaf organs in GBM with 0.3 mg·L⁻¹ BA, 0.01 mg·L⁻¹ Kin, 0.01 mg·L⁻¹ IBA, 0.3 g·L⁻¹ Pro, and 0.01 mg·L⁻¹ glutamine (Gln) accumulated 20.64 g fresh weight and 41.910 mg·g⁻¹ DW ginkgolic acids, representing a 4.93-fold increase over mother plants. This system enables large-scale Ginkgo biloba L. propagation and provides an in vitro strategy for producing medicinal compounds in endangered plants. Full article

Organic anion transporter 3 (OAT3), expressed at the basolateral membrane of kidney proximal tubule cells, facilitates the elimination of numerous metabolites, environmental toxins, and clinically important drugs. An earlier investigation from our laboratory revealed that OAT3 expression and transport activity can be upregulated by SUMOylation, a post-translational modification that covalently conjugates SUMO molecules to substrate proteins. Topotecan is a semi-synthetic derivative of the herbal extract camptothecin, approved by the FDA to treat several types of cancer. Ginkgolic acid (GA) is one of the major components in the extract of Ginkgo biloba leaves that has long been used in food supplements for preventing dementia, high blood pressure, and supporting stroke recovery. Both topotecan and GA have been shown to affect protein SUMOylation. In the current study, we tested our hypothesis that topotecan and GA may regulate OAT3 SUMOylation, expression, and transport function. Our data show that the treatment of OAT3-expressing cells with topotecan or GA significantly decreases the SUMOylation of OAT3 by 50% and 75%, respectively. The same treatment also led to substantial reductions in OAT3 expression and the OAT3-mediated transport of estrone sulfate, a prototypical substrate. Such reductions in cell surface expression of OAT3 correlated well with an increased rate of OAT3 degradation. Mechanistically, we discovered that topotecan enhanced the association between OAT3 and the SUMO-specific protease SENP2, a deSUMOylation enzyme, which contributed to the significant decrease in OAT3 SUMOylation. In conclusion, this study unveiled a novel role of topotecan and GA in inhibiting OAT3 expression and transport activity and accelerating OAT3 degradation by suppressing OAT3 SUMOylation. During comorbidity therapies, the use of topotecan or Ginkgo biloba extract could potentially decrease the transport activity of OAT3 in the kidneys, which will in turn affect the therapeutic efficacy and toxicity of many other drugs that are substrates for the transporter. Full article

Ginkgo biloba leaves (GBLs), which comprise many phytoconstituents, also contain a toxic substance named ginkgolic acid (GA). Our previous research showed that heating could decarboxylate and degrade GA into ginkgols with high levels of bioactivity. Several methods are available to measure GA in GBLs, but no analytical method has been developed to measure ginkgols and GA simultaneously. Hence, for the first time, an HPLC-DAD method was established to simultaneously determine GA and ginkgols using acetonitrile (0.01% trifluoroacetic acid, v/v) as mobile phase A and water (0.01% trifluoroacetic acid, v/v) as mobile phase B. The gradient elution conditions were: 0–30 min, 75–90% phase A; 30–35 min, 90–90% phase A; 35–36 min, 90–75% phase A; 36–46 min, 75–75% phase A. The detection wavelength of GA and ginkgol were 210 and 270 nm, respectively. The flow rate and injection volume were 1.0 mL/min and 50 μL, respectively. The linearity was excellent (R² > 0.999), and the RSD of the precision, stability, and repeatability of the total ginkgols was 0.20%, 2.21%, and 2.45%, respectively, in six parallel determinations. The recoveries for the low, medium, and high groups were 96.58%, 97.67%, and 101.52%, respectively. The limit of detection of ginkgol C13:0, C15:1, and C17:1 was 0.61 ppm, 0.50 ppm, and 0.06 ppm, respectively. The limit of quantification of ginkgol C13:0, C15:1, and C17:1 was 2.01 ppm, 1.65 ppm, and 0.20 ppm, respectively. Finally, this method accurately measured the GA and ginkgol content in ginkgo leaves and ginkgo tea products (ginkgo black tea, ginkgo dark tea, ginkgo white tea, and ginkgo green tea), whereas principal component analysis (PCA) was performed to help visualize the association between GA and ginkgols and five different processing methods for GBLs. Thus, this research provides an efficient and accurate quantitative method for the subsequent detection of GA and ginkgols in ginkgo tea. Full article

Ellagic acid, known for its various biological activities, is widely used. Ellagic acid from pomegranate peels is safe for consumption, while that from gallnuts is only suitable for external use. However, there is currently no effective method to confirm the source of ellagic acid. Therefore, this study establishes an analysis method using ultra-high-performance liquid chromatography–electrospray ionization–high-resolution mass spectrometry (UHPLC-ESI-HR-MS) to identify the components of crude ellagic acid extracts from pomegranate peels and gallnuts. The analysis revealed that there was a mix of components in the crude extracts, such as ellagic acid, palmitic acid, oleic acid, stearic acid, and 9(10)-EpODE. Furthermore, it could be observed that ellagic acid extracted from gallnuts contained toxic substances such as anacardic acid and ginkgolic acid (15:1). These components could be used to effectively distinguish the origin of ellagic acid from pomegranate peels or gallnuts. Additionally, a rapid quantitative analysis method using UHPLC-ESI-MS with multiple reaction monitoring (MRM) mode was developed for the quality control of ellagic acid products, by quantifying anacardic acid and ginkgolic acid (15:1). It was found that one of three ellagic acid health care products contained ginkgolic acid (C15:1) and anacardic acid at more than 1 ppm. Full article

The decolorization process plays a pivotal role in refining Ginkgo ketone ester by primarily eliminating ginkgolic acids, a toxic component. Presently, the conventional testing method involves sending samples for analysis, causing delays that impact formulation production. Hence, the development of a rapid process control method becomes imperative. This study introduces a swift detection approach for three ginkgolic acids during Ginkgo ketone ester’s decolorization. Initially, an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method assessed ginkgolic acid C13:0, ginkgolic acid C15:1, and ginkgolic acid C17:1 concentrations in 91 decolorized solution samples, establishing reference values. Subsequently, using a portable Raman spectrometer, Raman spectra of the decolorized liquid within the 3200–200 cm⁻¹ wavelength range were collected. Ultimately, employing partial least squares regression (PLSR) and ResNeXt50 deep learning algorithms, two quantitative calibration models correlated the ginkgolic acid content to Raman spectral data. Both models exhibited high predictive accuracy, with the ResNeXt50 model demonstrating superior performance. The prediction set correlation coefficients ($R_p^2$) for ginkgolic acid C13:0, ginkgolic acid C15:1, and ginkgolic acid C17:1 were 0.9962, 0.9971, and 0.9974, respectively, with root mean square error of prediction (RMSEP) values of 0.0144, 0.0130, and 0.0122 μg/mL. In contrast, the PLSR model yielded $R_p^2$ values of 0.9862, 0.9839, and 0.9480, with RMSEP values of 0.0273, 0.0305, and 0.0545 μg/mL for the three ginkgolic acids. The ResNeXt50 model not only showcased higher precision but also enhanced interpretability, as analyzed through gradient-weighted class activation mapping (Grad-CAM). The integration of Raman spectroscopy and the ResNeXt50 quantitative calibration model furnishes a real-time and precise approach to monitor ginkgolic acid content in the decolorized solution during Ginkgo ketone ester preparation. This significant advancement establishes a robust framework for implementing quality control measures in the decolorization process. Full article

Ginkgo biloba L. (Ginkgoacea) contains an abundance of beneficial compounds and has demonstrated positive clinical effects in the management of metabolic syndrome. Recent studies have emphasized its efficacy against type 2 diabetes mellitus (T2DM), including improvements in diabetic nephropathy and retinopathy. Particularly noteworthy are ginkgolic acid analogs, which have shown potential in combating T2DM by inhibiting protein tyrosine phosphatases (PTPs), facilitating glucose uptake, and influencing signaling pathways. In this study, we isolated six derivatives of ginkgolic acid from the MeOH extract of G. biloba leaves with the guidance of liquid chromatography–mass spectrometry (LC/MS). We determined the chemical structures of these isolated compounds as 2-hydroxy-6-(10′-hydroxypentadec-11′(E)-en-1-yl) benzoic acid (1), 2-hydroxy-6-(11′-hydroxypentadec-9′(E)-en-1-yl) benzoic acid (2), 2-hydroxy-6-tridecylbenzoic acid (3), 2-hydroxy-6-pentadecylbenzoic acid (4), 2-hydroxy-6-(12′-hydroxyheptadec-13′(E)-en-1-yl) benzoic acid (5), and 2-hydroxy-6-(11-hydroxyundecyl) benzoic acid (6) using NMR spectroscopic data and LC/MS analysis. To assess their potential for addressing T2DM, we subjected the isolated compounds (1–6) to tests measuring their inhibitory activity against six PTPs: PTPN11, PTPN2, PTP1B, DUSP9, PTPRS, and PTPN9. Among these compounds, compounds 3–5 displayed enzyme inhibition exceeding 90% against all six PTPs. In conclusion, ginkgolic acid derivatives, acting as PTP inhibitors relevant to insulin resistance, hold promise as potential therapeutic candidates for the prevention and treatment of T2DM. Full article

Drugs that cure COVID-19 have been marketed; however, this disease continues to ravage the world without becoming extinct, and thus, drug discoveries are still relevant. Since M^pro has known advantages as a drug target, such as the conserved nature of the active site and the absence of homologous proteins in the body, it receives the attention of many researchers. Meanwhile, the role of traditional Chinese medicine (TCM) in the control of epidemics in China has also led to a focus on natural products, with the hope of finding some promising lead molecules through screening. In this study, we selected a commercial library of 2526 natural products from plants, animals and microorganisms with known biological activity for drug discovery, which had previously been reported for compound screening of the SARS CoV-2 S protein, but had not been tested on M^pro. This library contains compounds from a variety of Chinese herbs, including Lonicerae Japonicae Flos, Forsythiae Fructus and Scutellariae Radix, which are derived from traditional Chinese medicine prescriptions that have been shown to be effective against COVID-19. We used the conventional FRET method for the initial screening. After two rounds of selection, the remaining 86 compounds were divided into flavonoids, lipids, phenylpropanoids, phenols, quinones, alkaloids, terpenoids and steroids according to the skeleton structures, with inhibition rates greater than 70%. The top compounds in each group were selected to test the effective concentration ranges; the IC₅₀ values were as follows: (−)–gallocatechin gallate (1.522 ± 0.126 μM), ginkgolic acid C15:1 (9.352 ± 0.531 μM), hematoxylin (1.025 ± 0.042 μM), fraxetin (2.486 ± 0.178 μM), wedelolactone (1.003 ± 0.238 μM), hydroxytyrosol acetate (3.850 ± 0.576 μM), vanitiolide (2.837 ± 0.225 μM), β,β–dimethylacrylalkannin (2.731 ± 0.308 μM), melanin (7.373 ± 0.368 μM) and cholesteryl sodium sulfate (2.741 ± 0.234μM). In the next step, we employed two biophysical techniques, SPR and nanoDSF, to obtain K_D/K_obs values: hematoxylin (0.7 μM), (−)–gallocatechin gallate (126 μM), ginkgolic acid C15:1 (227 μM), wedelolactone (0.9770 μM), β,β–dimethylacrylalkannin (1.9004 μM,), cholesteryl sodium sulfate (7.5950 μM) and melanin (11.5667 μM), which allowed better assessments of the binding levels. Here, seven compounds were the winners. Then, molecular docking experiments were specially performed by AutoDock Vina to analyze the mode of interactions within M^pro and ligands. We finally formulated the present in silico study to predict pharmacokinetic parameters as well as drug-like properties, which is presumably the step that tells humans whether the compounds are drug-like or not. Moreover, hematoxylin, melanin, wedelolactone, β,β–dimethylacrylalkannin and cholesteryl sodium sulfate are in full compliance with the “Lipinski” principle and possess reasonable ADME/T properties, they have a greater potential of being lead compounds. The proposed five compounds are also the first to be found to have potential inhibitory effects on SARS CoV-2 M^pro. We hope that the results in this manuscript may serve as benchmarks for the above potentials. Full article

Plant-derived natural compounds are widely used as alternative medicine in healthcare throughout the world. Ginkgolic acids, the phenolic compounds isolated from the leaves and seeds of Ginkgo biloba, are among the chemicals that have been explored the most. Ginkgolic acids exhibit cytotoxic activity against a vast number of human cancers in various preclinical models in vitro and in vivo. Additionally, the pharmacological activities of ginkgolic acids are also involved in antidiabetic, anti-bacteria, anti-virus, anti-fibrosis, and reno/neuroprotection. Autophagy as a highly conserved self-cleaning process that plays a crucial role in maintaining cellular and tissue homeostasis and has been proven to serve as a protective mechanism in the pathogenesis of many diseases, including neurodegenerative diseases, cancer, and infectious diseases. In this review, we surveyed the pharmacological activities of the major three forms of ginkgolic acids (C13:0, C15:1, and C17:1) that are linked to autophagic activity and the mechanisms to which these compounds may participate. A growing body of studies in last decade suggests that ginkgolic acids may represent promising chemical compounds in future drug development and an alternative remedy in humans. Full article

Ginkgols are active constituents from Ginkgo biloba L. (GB) and have pharmacological activities, such as antibacterial and antioxidant activities. In our previous report, only five ginkgols were separated. However, ginkgol C17:1 had two isomers, for which their separation, identification, and bioactivities have not yet been investigated. Hence, this research reports the successful isolation of six ginkgol homologs with alkyl substituents—C17:1-Δ¹², C15:1-Δ⁸, C13:0, C17:2, C17:1-Δ¹⁰, and C15:0—for the first time using HPLC. This was followed by the identification of their chemical structures using Fourier transform infrared (FTIR), ultraviolet (UV), gas chromatography and mass spectrometry (GC-MS), carbon-13 nuclear magnetic resonance (¹³C-NMR), and proton nuclear magnetic resonance (¹H-NMR) analysis. The results showed that two ginkgol isomers, C17:1-Δ¹² and C17:1-Δ¹⁰, were obtained simultaneously from the ginkgol C17:1 mixture and identified entirely for the first time. That aside, the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay showed that the six ginkgol homologs possessed significant antiproliferation effects against HGC and HepG2 cells. Furthermore, the ginkgols with unsaturated side chains (C17:2, C15:1-Δ⁸, C17:1-Δ¹², and C17:1-Δ¹⁰) exhibited more potent inhibitory effects than ginkgols with saturated side chains (C13:0, C15:0). In addition, unsaturated ginkgol C15:1-Δ⁸ showed the most potent cytotoxicity on HepG2 and HGC cells, of which the half-maximal inhibition concentrations (IC₅₀) were 18.84 ± 2.58 and 13.15 ± 2.91 μM, respectively. The IC₅₀ for HepG2 and HGC cells for the three unsaturated ginkgols (C17:1-Δ¹⁰, C17:2 and C17:1-Δ¹²) were ~59.97, ~60.82, and ~68.97 μM for HepG2 and ~30.97, ~33.81, and ~34.55 μM for HGC cells, respectively. Comparing the ginkgols’ structure–activity relations, the findings revealed that the position and number of the double bonds of the ginkgols with 17 side chain carbons in length had no significant difference in anticancer activity. Full article

Ginkgo tea and ginkgo wine are two familiar Ginkgo biloba leaf extract (GBE) drinks in the form of dietary supplements (DS) used for healthcare in east Asia. Nevertheless, a comprehensive evaluation of their safety and efficacy is still lacking. In this study, GBE drinks were prepared from naturally newly senescent yellow leaves (YL) and green leaves (GL) in autumn. Their total flavonoids, antioxidant capacity and prescribed ingredients were investigated. In brief, the proportions of total flavonoids, total flavonol glycosides (TFs), total terpene trilactones (TTLs) and ginkgolic acids in the GBE drinks all did not meet the standards of worldwide pharmacopoeias. Specifically, the levels of TFs in the ginkgo tea prepared from YL were significantly higher than that prepared from GL. Further analyses revealed a substandard ratio of isorhamnetin/quercetin and an accumulation of leaf-age-related compounds, which were both unqualified. The proportions of specific TTLs varied between the ginkgo tea and ginkgo wine, although no significant differences were detected in terms of the total levels of TTLs. Noticeably, numerous biflavones and thousands of times over the limiting concentration of ginkgolic acids, including newly identified types, were only detected in ginkgo wine. Finally, the use of the GBE drinks as DSs was comprehensively evaluated according to the acceptable daily intake. This study showed the limited healthcare effects of GBE drinks despite their powerful antioxidant capacity. Full article

A preliminary ethnopharmacological survey, achieved in French Polynesia, led to the collection of the most cited plants among 63 species used to treat “infectious” diseases, with a description of their medicinal uses. Bibliographical investigations and antimicrobial screening permitted the selection of the botanical species Syzygium malaccense (Myrtaceae) for phytochemical analysis. Leaves of Syzygium malaccense were usually used in mixture with rhizomes of Curcuma longa to treat infectious diseases such as cystitis. The methanolic plant extracts were tested in vitro with an agar microdilution method on 33 bacteria strains and 1 yeast to obtain their Minimal Inhibitory Concentration (MIC), and cytotoxicity against HepG2 cells were evaluated. Antimicrobial synergistic effects of methanolic plant extracts from leaves of Syzygium malaccense and rhizomes from Curcuma longa were also evaluated. The bio-guided isolation of leaf extract from Syzygium malaccense led to the identification of seven alkyl-salicylic acids (anacardic acids or ginkgolic acids C15:0, C15:1, C17:0, C17:1, C17:2, C17:3 and C19:1) described for the first time in this species. All compounds were tested against Staphylococcus aureus (18.75 < MIC < 75.0 µg/mL), Streptococcus pyogenes (2.34 < MIC < 18.75 µg/mL) and Pseudomonas aeruginosa (MIC = 150 µg/mL), and their structure–activity relationships were discussed. The methanolic extract and salicylic derivatives from S. malaccense showed an interesting antimicrobial activity against Gram+ bacteria, without toxicity on hepG2 cells at 400 μg/mL. Moreover, these antibacterial compounds have already been studied for their anti-inflammatory activity, which supports the therapeutic interest of S. malaccense against infectious diseases. Full article

Ginkgolic acid (C13:0) (GA), isolated from Ginkgo biloba, is a potential therapeutic agent for type 2 diabetes. A series of GA analogs were designed and synthesized for the evaluation of their structure–activity relationship with respect to their antidiabetic effects. Unlike GA, the synthetic analog 1e exhibited improved inhibitory activity against PTPN9 and significantly stimulated glucose uptake via AMPK phosphorylation in differentiated 3T3-L1 adipocytes and C2C12 myotubes; it also induced insulin-dependent AKT activation in C2C12 myotubes in a concentration-dependent manner. Docking simulation results showed that 1e had a better binding affinity through a unique hydrophobic interaction with a PTPN9 hydrophobic groove. Moreover, 1e ameliorated palmitate-induced insulin resistance in C2C12 cells. This study showed that 1e increases glucose uptake and suppresses palmitate-induced insulin resistance in C2C12 myotubes via PTPN9 inhibition; thus, it is a promising therapeutic candidate for treating type 2 diabetes. Full article