Introduction

Medicinal plants play a crucial role in the development of pharmaceutical and cosmetic products, serving as a rich source of bioactive compounds. These natural compounds, including flavonoids, terpenoids, and phenolic acids, are known for their potent antioxidant and anti-inflammatory properties.1,2 Flavonoids, as one of the largest groups of polyphenolic compounds, have been extensively studied for their ability to scavenge free radicals and regulate oxidative stress. Oxidative stress occurs when an imbalance between free radical production and antioxidant defenses leads to cellular damage, lipid peroxidation, and inflammation.3 This condition is a major contributor to chronic diseases such as cancer, cardiovascular disorders, and neurodegenerative diseases.4 Antioxidant compounds from plants have been shown to mitigate these effects by enhancing the body’s endogenous defense systems.5 Moreover, flavonoids and terpenoids are increasingly recognized for their enzyme inhibitory properties, offering therapeutic potential against various pathophysiological processes.6,7 These attributes highlight the significance of exploring phytochemicals for their pharmacological and cosmetic applications.

Artocarpus heterophyllus, commonly known as jackfruit, is a tropical plant that has been traditionally used for its nutritional and medicinal properties. Various parts of the plant, particularly its leaves, have been utilized in treating skin disorders, inflammation, and oxidative stress-related conditions.8,9 The leaves of Artocarpus heterophyllus are rich in flavonoids and terpenoids, which are bioactive compounds with diverse therapeutic benefits. Research has shown that these compounds exhibit antioxidant and anti-inflammatory activities, supporting the traditional use of the plant in addressing oxidative stress.10 Furthermore, terpenoids and phenolic acids found in the leaves are associated with regulating key enzymes involved in inflammation and tissue remodeling. In addition to their therapeutic roles, Artocarpus heterophyllus extracts have potential applications in cosmetics, particularly in controlling pigmentation and improving skin health.11,12 The plant’s ability to address both health and cosmetic concerns underscores its versatility as a natural remedy. Exploring the phytochemical composition of Artocarpus heterophyllus leaves is essential to understanding their bioactive potential.13

This study focuses on analyzing the bioactive compounds present in Artocarpus heterophyllus leaf extracts obtained using ethyl acetate and ethanol solvents. The choice of these solvents is based on their polarity, which influences the range and efficiency of compound extraction. Techniques such as Thin Layer Chromatography (TLC) and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) were employed to identify and characterize the major compounds in the extracts. The antioxidant activity of the extracts was evaluated to assess their potential in mitigating oxidative stress, a key factor in chronic diseases and aging.14 Additionally, the tyrosinase inhibitory activity of the extracts was studied for its relevance in cosmetic applications, particularly in skin pigmentation control.15,16 The study also explores the correlation between the chemical profiles of the extracts and their observed bioactivities. By understanding these properties, the research seeks to highlight the therapeutic and cosmetic applications of Artocarpus heterophyllus extracts. These findings are expected to provide insights into the plant’s potential as a source of natural bioactive compounds.

By integrating traditional knowledge with scientific research, this study aims to establish the significance of Artocarpus heterophyllus leaf extracts in health and skincare. The investigation highlights the role of solvent selection in optimizing the extraction of bioactive compounds. The study’s focus on antioxidant and tyrosinase inhibitory activities aligns with current trends in developing natural alternatives for therapeutic and cosmetic applications. These efforts contribute to the growing interest in plant-based solutions for managing oxidative stress and pigmentation-related concerns.17,18Artocarpus heterophyllus, with its rich phytochemical profile, emerges as a promising candidate for further exploration in these fields. Utilizing advanced analytical techniques, including LC-MS/MS, molecular docking, and molecular dynamics (MD) simulations, the study provides a comprehensive understanding of the phytochemical composition and its bioactivities. These findings not only validate the ethnopharmacological uses of Artocarpus heterophyllus but also support its relevance in modern therapeutic and cosmetic innovations (Video S1). Future research will focus on clinical evaluations and optimizing formulations to enhance bioavailability and efficacy, paving the way for its integration into health and skincare solutions.

Material and Methods General

TLC analyses were conducted using 20×20 cm aluminum sheets coated with silica gel 60 F254 (Sigma-Aldrich, St. Louis, USA) and 0.25 mm silica gel G-25 UV254 pre-coated glass TLC plates (Sigma-Aldrich, St. Louis, USA), suitable for both vacuum column chromatography and preparative TLC procedures. Terpenoids and steroids were visualized on TLC plates by applying Liebermann-Burchard reagent as a staining agent. For compound profiling, a UHPLC Vanquish system coupled with a Q Exactive Plus Orbitrap High-Resolution Mass Spectrometer (Thermo Scientific, Waltham, MA, USA) was used to perform LC-MS/MS analysis. NMR data were collected using a Bruker Avance III™ HD 600 MHz NMR spectrometer (Bruker BioSpin GmbH, Rheinstetten, Germany), with ^1H spectra recorded at 500 MHz and ^13C spectra at 150 MHz, using TMS as the reference standard in deuterated chloroform, and chemical shifts reported in δ (ppm). Antioxidant testing utilized DPPH radical scavenging assays, where a microplate reader recorded absorbance at 517 nm. The assay was performed in 70% ethanol, with Trolox as the standard, and results were reported as µmol Trolox equivalents (TE) per gram of sample. For the tyrosinase inhibition assay, absorbance was measured at 475 nm using a multi-well plate reader. Solutions were prepared with L-DOPA in a 50 mM phosphate buffer (pH 6.8), and kojic acid was used as a positive control for reference in evaluating enzyme inhibition activity.

Plant Material

The Artocarpus heterophyllus leaves were collected from Kota Bandung, West Java, Indonesia, in August 2024. The plant specimen was formally identified and authenticated at the Herbarium Bandungense, housed within the School of Life Sciences and Technology, Bandung Institute of Technology. The identification process was completed under voucher number 8366/IT1.C11.2/TA.00/2024, and the specimen was subsequently deposited in the herbarium’s collection for documentation and future reference. The identification and deposition process were officially validated by the Deputy Dean of Resources, Angga Dwiartama, S.Si., M.Si., Ph.D., ensuring the credibility and accuracy of the botanical identification.

Extraction and Isolation

The leaves of Artocarpus heterophyllus (1.6 kg) were extracted using the maceration method with ethyl acetate (16 L) as the solvent over a period of 3–5 days. The resulting filtrate was evaporated to yield a viscous ethyl acetate extract, with a total weight of 6.078 g for batch 1 and 25.573 g for batch 2. This extract was then monitored using Thin Layer Chromatography (TLC) to confirm the presence of the desired compounds. Additionally, a separate extraction was performed using ethanol as the solvent. A total of 1 kg of Artocarpus heterophyllus leaves was extracted in 10 L of ethanol using the maceration method for 3–5 days. The ethanol filtrate was evaporated to yield a viscous ethanolic extract, resulting in 15.8 g for batch 1 and 26.4579 g for batch 2. This ethanolic extract was also monitored using TLC to identify the compound components. TLC monitoring revealed several fractions with specific Rf patterns, which were further isolated. The identified compounds exhibited distinct characteristics in TLC, suggesting the presence of compound groups such as flavonoids, phenolic acids, and terpenoids/steroids.19,20 In the ethyl acetate extract, a fraction with a specific Rf value of 0.52 (brown color with sulfuric acid-methanol) was identified and collected as a colorless powder. In the ethanol extract, a fraction with an Rf value of approximately 0.48 (brown color with sulfuric acid-methanol) was isolated and collected in the form of a colorless powder. All isolated compounds from both extracts were further analyzed using LC-MS/MS for further characterization.

LC-MS/MS Analysis

Each isolated compound (1 mg) was dissolved in 1 mL of methanol (LC-MS grade, Chromasolv®). A 1-microliter Artocarpus heterophyllus leaves extract aliquot was injected into the column. A gradient elution was performed using 0.1% formic acid (v/v) in water alongside acetonitrile with 0.1% formic acid (v/v), starting with a ratio of 95:5, shifting to 60:40 from 1.00 to 8.00 minutes, then 0:100 from 8.00 to 13.00 minutes, and concluding with 95:5. The flow rate was maintained at 0.3 mL/min. Instrument settings included: acquisition time of 0.00–16.00 minutes, mass range of 50.00–1200.00 m/z, scan interval of 0.100 s, low CE of 6 eV, high CE ranging from 10–40 eV, cone voltage at 30 V, collision energy at 6 eV, acquisition mode ESI (+), capillary voltage at 2 kV, source temperature set to 120 °C, desolvation temperature at 500 °C, cone gas flow at 50 L/h, desolvation gas flow at 1000 L/h, sample temperature set to 20 °C, and column temperature at 40 °C.21 Data were processed with UNIFI software (version 1.8, Waters Corporation, Milford, MA, USA), utilizing a screening solution workflow to facilitate automated data handling and positive identifications.22 The results were compared against a database containing over 1200 compounds, cross-referenced by chemical structure, molecular formula, and molecular weight from multiple online resources.

Multi Targeted Molecular Docking Study

For the molecular docking analysis, various crystal structures were utilized, including human tyrosinase-related protein 1 (PDB ID: 5M8Q), Melanocyte Inducing Transcription Factor (MITF) (PDB ID: 5T3Q), Matrix Metalloproteinase-1 (MMP-1, PDB ID: 1HFC), Matrix Metalloproteinase-2 (MMP-2, PDB ID: 1HOV), Matrix Metalloproteinase-8 (MMP-8, PDB ID: 5H8X), Matrix Metalloproteinase-9 (MMP-9, PDB ID: 2OW1), Matrix Metalloproteinase-13 (MMP-13, PDB ID: 2OZR), TGF-Beta receptor type 1 kinase (PDB ID: 6B8Y), fibroblast collagenase (PDB ID: 1CGL), and elastase (PDB ID: 3HGP).23–31 The selected receptor crystal structures were chosen based on their roles in oxidative stress, inflammation, and pigmentation-related disorders, aligning with the bioactivities of the tested compounds. Proteins underwent refinement via the Protein Preparation Wizard in Schrödinger’s Maestro 2020–3 software (Schrödinger, New York, NY, USA), where missing hydrogens were added, and partial charges were assigned using the OPLS_2005 (Optimized Potentials for Liquid Simulations 2005) forcefield.32,33 Proteins were prepped in a restrained minimization state with hydrogens and heavy atoms. The 2D chemical structures of compounds from Artocarpus heterophyllus leaves extract, identified through LC-MS/MS, were converted into 3D forms using the LigPrep Module in Maestro Schrödinger 2020–3 with OPLS_2005, adjusting pH to 7.4 through Epik. LigPrep ensured appropriate protonation states, tautomeric forms, and ionization, with bond orders validated for accuracy. To define the docking region, the grid box was aligned by selecting the native ligand from the protease receptor to centralize docked compounds within the same spatial dimensions as the binding region. The docking procedure was performed in extra precision (XP) mode through Glide, applying the OPLS_2005 forcefield under flexible ligand and fixed receptor parameters. Each ligand’s efficacy as a receptor inhibitor was assessed by scoring docked poses using the Molecular Mechanics-Generalized Born Surface Area (MM-GBSA) methodology.34

Multi Targeted Molecular Dynamics Simulation

GROMACS 2024.4 was utilized to perform molecular dynamics (MD) simulations on the protein-Artocarpin and protein-Sitosterol complexes, each simulated over 50 ns.35,36 A dodecahedral unit cell was defined with a 1 nm buffer between the protein surface and box edge, solvated using the Simple Point Charge (SPC) water model. The CHARMM36 (Chemistry at HARvard Macromolecular Mechanics 36) was selected for target topologies, and the system underwent energy minimization with the steepest descent integrator for 50,000 steps, targeting a force convergence threshold of 1000 kcal/mol/nm.37 Following this, each protein-ligand complex was equilibrated under both NVT (constant number of particles, volume, and temperature) and NPT (constant number of particles, pressure, and temperature) conditions for 1000 ps, with temperature and pressure stabilization set to 300 K and 1 bar using the Berendsen thermostat and Parrinello-Rahman barostat, respectively. MD simulations were conducted for 50 ns with coordinate snapshots saved every 2 fs, balancing between runtime efficiency and data capture. The stability observed in fluctuation graphs confirmed successful equilibration. GROMACS analytical tools were employed for structural and conformational analyses across systems, while MM-PBSA (Molecular Mechanics-Poisson Boltzmann Surface Area) calculations were used to estimate binding free energies from the snapshots.38

DPPH Radical Scavenging Activity

The DPPH radical scavenging activity was assessed by combining 100 µL of the Artocarpus heterophyllus leaves extract with 100 µL of DPPH solution (0.1 mM in 70% ethanol) and incubating the mixture in the dark at 25–27 °C for 30 minutes. Distilled water was used as a control and as a substitute for the sample. Absorbance was measured at 517 nm using a microplate reader, and scavenging activity was calculated as follows: Scavenging activity (%) = [(Acontrol − Asample) / Acontrol] × 100, where Acontrol is the absorbance of the DPPH solution alone, and Asample is the absorbance of the sample mixed with DPPH.39 The activity was reported as IC50, representing the concentration of the sample needed to inhibit 50% of the initial DPPH concentration. The IC50 value was obtained from a linear regression graph, and results were expressed in µmol Trolox equivalents (TE) per gram of sample for each fraction tested.

Tyrosinase Enzyme Inhibitory Activity

The tyrosinase inhibition activity of Artocarpus heterophyllus leaves extract was determined by preparing a reaction mixture in a multi-well plate. A 3.2 mM L-DOPA solution in 50 mM phosphate buffer (pH 6.8) was prepared. Then, 10 µL of various concentrations of Artocarpus heterophyllus leaves extract was diluted with 40 µL of 50 mM K₃PO₄ buffer (pH 6.8) and combined with 70 µL of tyrosinase solution (150 units/mL). This mixture was incubated for 10 minutes. Afterward, 80 µL of the L-DOPA solution (3.2 mM) was added to each well and left to incubate for another 10 minutes. The absorbance was read at 475 nm using a multi-well plate reader. Kojic acid was used as a positive control to benchmark inhibition. Tyrosinase inhibition activity was calculated with the following formula: Tyrosinase inhibitory activity (%) = × 100, where Ablank is the absorbance of deionized water, Acolour is the absorbance of the sample mixed with phosphate buffer, Acontrol represents the absorbance of the buffer-tyrosinase-L-DOPA mixture without the sample, and Asample is the absorbance from the reaction solution.40 The IC50 values were calculated similarly to those in the DPPH analysis.

Results Extraction of Artocarpus Heterophyllus Leaves

The extraction of Artocarpus heterophyllus leaves using ethyl acetate and ethanol through maceration yielded different concentrations of bioactive compounds. Ethyl acetate extraction produced 0.38% yield (6.078 g from 1.6 kg of leaves), while ethanol extraction achieved a higher yield of 1.58% (15.8 g from 1 kg of leaves). Thin Layer Chromatography (TLC) and LC-MS/MS analyses revealed that both extracts were dominated by flavonoids and terpenoids/steroids, though in varying proportions. Flavonoids constituted 50% of the ethyl acetate extract and 35% of the ethanol extract, while terpenoids/steroids made up 25% and 30%, respectively (Figure 1). Phenolic acids were also present in moderate amounts, with a slightly higher concentration in the ethanol extract. Hydrocarbons and other compounds appeared in minimal quantities across both extracts, indicating that the primary components were flavonoids and terpenoids/steroids. These findings underscore the selective extraction capabilities of ethyl acetate and ethanol, emphasizing the influence of solvent polarity. Both extracts thus show a high potential for yielding bioactive compounds, with ethanol demonstrating a broader range of extractable compounds.

Figure 1 Percentage composition of compound categories in ethyl acetate and ethanol extracts of Artocarpus heterophyllus leaves.

The prominent presence of flavonoids and terpenoids/steroids in both ethyl acetate and ethanol extracts suggests significant potential for isolating these bioactive compounds from Artocarpus heterophyllus leaves. Flavonoids are widely known for their antioxidant properties, while terpenoids and steroids have various therapeutic applications, highlighting the extracts’ value in pharmaceutical contexts. This study recommends using liquid-liquid extraction with solvents of varying polarity, such as n-hexane and water, to further enhance isolation of these compounds by optimizing selectivity based on solubility. Such targeted extraction methods may yield purer fractions, allowing for more effective pharmacological studies on individual compounds. Additionally, the presence of phenolic acids, although moderate, contributes to the overall antioxidant potential of the extracts.41,42 These results emphasize the leaves of Artocarpus heterophyllus as a viable source of beneficial phytochemicals, particularly flavonoids and terpenoids.

Isolation and LC-MS/MS Identification From Isolates of Artocarpus Heterophyllus Leaves

The ethyl acetate and ethanol extracts of Artocarpus heterophyllus leaves were analyzed to identify their compound composition, focusing on key compounds based on retention time, molecular mass, and molecular formula (Table 1, Figure 2). In the ethyl acetate extract, Cintramide emerged as the most abundant compound, comprising 46.61% of the extract at a retention time of 7.48 minutes, indicating its strong affinity for ethyl acetate. Other significant compounds included Licoflavone C and 3,4,5-trimethoxy cinnamic acid, with compositions of 19.14% and 10.53%, respectively. These compounds, especially flavonoids and cinnamic acid derivatives, are notable for their antioxidant and anti-inflammatory properties, which suggest that the ethyl acetate extract may possess significant bioactivity. The presence of smaller amounts of compounds like Sitosterol and Artocarpin, though less abundant, adds to the diverse composition of the extract. Each of these compounds was carefully identified using TLC and LC-MS/MS, ensuring accurate classification. The predominance of these bioactive molecules in the ethyl acetate extract makes it a valuable source for further isolation studies.

Table 1 Compounds Identified by LC-MS/MS From the Isolates of Artocarpus Heterophyllus Ethyl Acetate and Ethanol Extracts

Figure 2 LC-MS/MS chromatogram of compounds identified from the isolates of Artocarpus heterophyllus ethyl acetate and ethanol extracts.

In contrast, the ethanol extract exhibited a unique chemical profile with several complex compounds that differed from those found in the ethyl acetate extract. The primary compound in the ethanol extract was (2s)-2-[(4-phenyl)formamido]pentanedioic acid, accounting for 44.14% of the total composition and detected at a retention time of 8.79 minutes. This compound’s presence in high concentration underscores the ethanol extract’s potential for therapeutic applications, particularly due to the bioactive properties often associated with such structures. Other prominent compounds included Licoflavone C (14.57%) and (2s)-3--2-methylpropanoic acid (8.48%), which contribute to the extract’s diversity. Smaller amounts of compounds, such as Asterric acid and Artocarpanone, added further complexity to the extract’s profile, indicating a wide range of potential bioactivities. The varied composition of the ethanol extract suggests a strong potential for therapeutic use.

In addition, the differences between the ethyl acetate and ethanol extracts highlight the influence of solvent choice on extraction efficiency and compound profile.43,44 Ethyl acetate appears to be more effective at extracting simple bioactive compounds, with Cintramide, Licoflavone C, and cinnamic acid derivatives making up the bulk of the extract. Meanwhile, ethanol effectively extracted larger, more complex molecules such as (2s)-2-[(4-phenyl)formamido]pentanedioic acid, along with other unique flavonoids and phenolic compounds. These differences in compound profiles suggest that both extracts hold promise for various therapeutic applications, with ethyl acetate potentially targeting antioxidant and anti-inflammatory effects, while ethanol may focus on more complex pharmacological activities. Both extracts’ bioactive potential, especially as natural sources of antioxidants and therapeutic agents.

Multi Targeted Molecular Docking Study

The molecular docking simulations of compounds identified from Artocarpus heterophyllus extracts provide valuable insights into their potential as enzyme inhibitors targeting fibroblast collagenase, elastase, MITF, MMP-1, and MMP-2. Table 2 reveals that several compounds, particularly Cycloaltilisin 7 and Sitosterol, exhibited strong binding affinities with low MMGBSA binding energies, suggesting stable interactions with these enzymes. Cycloaltilisin 7 stood out with binding energies as low as −10.71 kcal/mol, indicating its robust inhibitory potential across multiple receptors. Similarly, Sitosterol showed consistent low binding energy values, especially −10.71 kcal/mol, positioning it as a promising candidate for enzyme inhibition. Artocarpin and Atocarpesin displayed moderate binding affinities, with binding energies around −8.87 and −8.72 kcal/mol, suggesting potential effectiveness in enzyme inhibition. Other compounds, like Licoflavone C-1 and Artocarpanone-3, exhibited notable binding energies, implying they may contribute to inhibition, although with slightly weaker affinities than Cycloaltilisin 7 and Sitosterol. This diversity of binding affinities across compounds highlights the broad bioactive potential of Artocarpus heterophyllus extracts. The results suggest a promising foundation for further exploration of these compounds in enzyme inhibition applications.

Table 2 Molecular Docking Simulations of Metabolites Identified in Artocarpus Heterophyllus Extracts on Fibroblast Collagenase, Elastase, MITF, MMP-1, and MMP-2 Predicted Potential Inhibitory Compounds

Table 3 provides additional insights into the inhibitory potential of compounds from Artocarpus heterophyllus ethyl acetate and ethanol extracts, with Sitosterol and Cycloaltilisin 7 showing exceptional binding affinities. Sitosterol demonstrated the strongest interaction, with binding energies as low as −13.00 kcal/mol, highlighting its high affinity and potential as a powerful enzyme inhibitor. Cycloaltilisin 7 also maintained low binding energies across multiple targets, reaching −11.19 kcal/mol, which reinforces its inhibitory capability. Other compounds, such as Artocarpanone-3 and Licoflavone C-1, displayed binding energies of −9.88 and −10.62 kcal/mol, suggesting they could contribute to enzyme inhibition, although with slightly less potency. Compounds like (2s)-2-[(4-phenyl)formamido]pentanedioic acid and additional phenolic acids. These findings underscore the complementarity of extraction methods in isolating distinct phytochemical profiles. Ethanol extraction proved especially effective, yielding a more diverse profile of bioactive compounds, including phenolic acids, which are essential in managing oxidative stress. Artocarpin, one of the primary compounds identified in the ethanol extract, and Sitosterol, found in lower concentrations in the ethyl acetate extract, are key contributors to the therapeutic value of the extract. Artocarpin’s recognized antioxidant and anti-inflammatory properties support its potential in pharmaceutical applications targeting inflammatory and oxidative stress-related diseases.53,54 Meanwhile, Sitosterol’s structural qualities position it as an ideal candidate for enzyme inhibition.55 This solvent-selective approach highlights how extraction methods influence bioactive composition, guiding targeted extraction of specific therapeutic compounds. By focusing on Artocarpin and Sitosterol, the study demonstrates how Artocarpus heterophyllus leaf extracts may serve as a source of valuable compounds for health applications.

Molecular docking simulations further emphasized the enzyme inhibition potential of Artocarpin and Sitosterol, showing their strong binding affinities with enzymes such as MMP-1, MMP-2, and MMP-13. These enzymes are key players in tissue remodeling and inflammation, processes often overactive in chronic inflammatory conditions and certain cancers. Sitosterol’s notably low binding energy with MMP-13 suggests a stable interaction, positioning it as a potent inhibitor with potential to disrupt MMP-related pathological pathways. Artocarpin, though slightly less potent than Sitosterol, still displayed significant binding affinity with MMP enzymes, indicating that it could modulate enzyme activity in diseases involving excessive MMP activity, such as chronic inflammation, arthritis, and cancer metastasis. To verify these initial findings, molecular dynamics (MD) simulations were conducted, providing a more comprehensive picture of how Artocarpin and Sitosterol behave over time when bound to their targets. Low Root Mean Square Deviation (RMSD) values for Artocarpin suggest a stable interaction with MMPs, as minimal structural deviation implies a steady binding. Additionally, both compounds exhibited low Radius of Gyration (Rg) values, indicating compact binding conformations essential for effective enzyme inhibition.56 The MD results, therefore, reinforce the potential of Artocarpin and Sitosterol as stable, natural MMP inhibitors, laying a foundation for their further exploration as therapeutic agents.

Beyond enzyme inhibition, the antioxidant and tyrosinase inhibitory activities of Artocarpin and Sitosterol offer additional therapeutic and cosmetic benefits. Artocarpin, particularly in the ethanol extract, displayed significant antioxidant activity, effectively scavenging free radicals, though slightly less potent than ascorbic acid. This antioxidant capacity suggests its utility in managing oxidative stress, a contributing factor in numerous chronic conditions, including aging-related diseases and cancer. Sitosterol, on the other hand, exhibited impressive tyrosinase inhibition, even outperforming kojic acid—a standard tyrosinase inhibitor widely used in cosmetic applications. Tyrosinase inhibition is crucial for managing melanin production and hyperpigmentation, making Sitosterol valuable for formulations aimed at achieving even skin tone. The polar nature of ethanol likely enhanced the extraction of tyrosinase-inhibiting compounds, making the ethanol extract particularly promising for pigmentation control in cosmetic applications.57,58 These results underscore the versatility of Artocarpin and Sitosterol, suggesting that they could be used in both skincare and health products. For Artocarpin, studies have shown limited solubility in aqueous environments, with an estimated bioavailability of less than 20%, which may restrict its effectiveness unless advanced delivery systems such as nanoparticles or encapsulation are applied to enhance its absorption. Sitosterol, on the other hand, demonstrates moderate bioavailability, typically ranging between 30% and 40% in oral formulations, though its hydrophobic nature necessitates solubilization techniques to improve systemic uptake. Addressing these bioavailability challenges is essential to maximize their therapeutic and cosmetic potential, paving the way for their incorporation into innovative and effective formulations.

Additionally, in vivo studies are essential to evaluate the impact of Artocarpin and Sitosterol under physiological conditions, focusing on oxidative stress mitigation, enzyme inhibition, melanin regulation, and anti-inflammatory effects. Advanced formulation strategies, such as nanoemulsions or liposomal delivery systems, could significantly enhance the bioavailability and stability of these compounds, ensuring prolonged activity and precise delivery to target sites. These technologies can also reduce potential side effects by minimizing off-target interactions, which is critical for their application in pharmaceutical and cosmetic products.59,60 The ability to combine these advanced formulations with natural bioactive compounds like Artocarpin and Sitosterol offers a promising avenue for creating more effective treatments. Furthermore, validating these compounds through clinical trials will provide definitive evidence of their therapeutic and cosmetic potential, especially in addressing age-related conditions, pigmentation disorders, and inflammatory diseases. Ultimately, this study lays the groundwork for leveraging Artocarpus heterophyllus leaf extracts in modern health and skincare solutions.

Conclusion

In conclusion, this study highlights the significant bioactive potential of Artocarpus heterophyllus leaf extracts for therapeutic and cosmetic applications. Artocarpin and Sitosterol, the primary bioactive compounds identified, exhibit notable antioxidant and tyrosinase inhibition activities. Ethanol extracts demonstrated stronger tyrosinase inhibition with an IC50 value of 177.24 ppm, while ethyl acetate extracts showed superior antioxidant activity with an IC50 value of 117.64 ppm. Molecular docking and molecular dynamics (MD) simulations further validated the potential of these compounds, with Artocarpin showing stable binding interactions with MMP-13, underscoring its role as a therapeutic agent targeting enzyme inhibition. These findings reveal the potential of Artocarpus heterophyllus leaf extracts in addressing oxidative stress, inflammation, and hyperpigmentation-related disorders. Future research should focus on conducting comprehensive in vivo studies to confirm these bioactivities under physiological conditions and optimizing advanced delivery systems, such as nanoemulsions or liposomal formulations, to enhance the bioavailability and stability of these compounds. By integrating these approaches, Artocarpus heterophyllus leaf extracts could pave the way for innovative plant-based solutions in modern therapeutic and cosmetic applications.

Acknowledgments

This work was supported by a grant from the Ministry of Education, Culture, Research, and Technology (Kemendikbudristek) of Indonesia under Contract Number: 305/B.04/Rek/VI/2024. We extend our gratitude for their funding and support, which made this research possible.

Disclosure

Dr. Lelly Yuniarti, S.Si., M.Kes; Dr. Maya Tejasari, dr., M.Kes; Dr. R. Anita Indriyanti, dr., M.Kes; and apt. Taufik Muhammad Fakih, S.Farm., M.S.Farm report a patent “Optimization of Extraction and Identification Protocols for Bioactive Compounds in Jackfruit Leaves (Artocarpus heterophyllus) Utilizing High-Resolution Liquid Chromatography Mass Spectrometry (LC-HRMS) for Anti-Aging Application [S00202410299]” issued. The authors declare no other conflicts of interest regarding the publication of this article.

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