Nungruthai Suphrom

The inhibition of steroid 5-alpha reductase (S5AR), a key mechanism for managing dihydrotestosterone-dependent conditions, has been demonstrated in teak (Tectona grandis L.f.) leaf extracts. Our recent clinical study confirmed the effectiveness of a hair growth formulation containing teak leaf extract in males with androgenic alopecia. However, significant variability in S5AR inhibitory activity among teak leaf samples from different regions underscores the need for quality control of raw materials. This study applied a metabolomics approach to investigate the influence of leaf age, harvesting period, and geographic origin on chemical composition and S5AR inhibitory activity, as well as to identify active S5AR inhibitors. Geographic origin emerged as the primary determinant of variations in chemical profiles and S5AR inhibitory activity. Using orthogonal partial least squares analysis, six diterpenoid S5AR inhibitors were identified, including four compounds reported for the first time as S5AR inhibitors: rhinocerotinoic acid, 7-oxo-8-labden-15-oic acid, 8-hydroxy-labd-13-en-15-oic acid, and a novel diterpene, 7-hydroxy-labd-8,13-dien-15-oic acid. These findings highlight the potential of metabolomics as a powerful tool for discovering bioactive compounds and optimizing raw material selection. By prioritizing proven geographic sources, consistent bioactivity can be achieved, supporting the therapeutic potential of teak leaves in managing S5AR-related conditions.

Context: Marine sponges from the genus Aaptos produce compounds, often alkaloids, with a wide range of bioactivities. Aaptos suberitoides exhibits anticancer, antioxidant, antibacterial proteasome inhibitory, and receptor activator of NF-kappaB ligand (RANKL) inhibitory activities. Alkaloids from marine sponges have shown potency as cholinesterase inhibitors, one of the targets in Alzheimer’s disease treatment. Aims: To isolate, identify, and investigate the potency of alkaloids from the water fraction of marine sponge A. suberitoides as cholinesterase inhibitors. Methods: Bioassay-guided isolation was employed to obtain active alkaloids. The cholinesterase inhibitory assay was conducted based on Ellman’s method with slight modification. Identification of isolated compounds was carried out based on NMR and MS data. The identified compounds were also subjected to molecular docking analysis as well as an ADMET study. Results: Bioassay-guided isolation of water fraction yielded aaptamine (1) and a mixture containing five benzo[de][1,6]-naphthyridine derivatives, identified based on NMR and LC-MS/MS spectroscopy. Aaptamine (1) inhibited both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) with IC50 values of 0.23 and 1.38 µg/mL, respectively. Likewise, the benzo[de][1,6]-naphthyridine mixture of N-demethylaaptanone (2), aaptanone (3), demethylaaptamine (4), aaptosine (5) and 8,9,9-trimethoxy-9H-benzo[de][1,6]-naphthyridine (6) recorded IC50 values of 0.39 and 1.61 µg/mL, respectively. Computational molecular docking analysis calculated the S-score and RMSD values of aaptamine (1) to be the best compared to galantamine for both AChE (PBD: 4EY6) and BChE (PDB: 4BDS) with values of -6.65 kcal/mol, 0.57 Å and -5.98 kcal/mol, 1.05 Å, respectively. Conclusions: These findings suggest that benzo[de][1,6]-naphthyridine alkaloids from A. suberitoides can be good candidates as cholinesterase inhibitors.

The leaf extract of Tectona grandis L.f. has shown potential as a 5α-reductase inhibitor, with two bioactive markers, namely (+)-eperua-8,13-dien-15-oic acid (1) and (+)-eperua-7,13-dien-15-oic acid (2), used for extract standardization. The purpose of this research was to investigate the in vitro skin penetration behavior of 1 and 2 in T. grandis leaf ethanolic extract solution and ready-to-use extract in propylene glycol (PG), and secondly, to determine their physicochemical properties, including partition coefficients and solubility. The appropriate vehicle for the in vitro skin penetration study was evaluated using the shake-flask method. The in vitro skin penetration study was conducted using the Franz diffusion cell model, and the amounts of the two active compounds in the extracts were analyzed using the HPLC method. Compounds 1 and 2 showed poor solubility in distilled water, whereas their solubility in HEPES buffer with 2% w/v of Tween 20 was significantly greater. The partition coefficient (log P_o/w) value for 1 was 5.77 ± 0.07, and for 2, it was 5.66 ± 0.02, indicating that both compounds are hydrophobic. After 24 h of an in vitro skin penetration study, 1 in both extracts showed significantly higher cumulative amounts (%) compared to 2. These findings suggest that 1 is more hydrophobic and readily penetrates the stratum corneum. When a PG enhancer was added, high cumulative amount trends of 1 and 2 in the ethanolic extract and extract in PG in the receiver compartment were detected after 24 h. These studies provide important insights that will guide the further development of products with T. grandis extracts for treating hair loss.

The study aimed to investigate the antioxidant and cholinesterase inhibitory activities of the leaves and stems of Artocarpus sericicarpus and to analyse the phenolic compounds in the extracts. The modified Ellman’s method was used to determine the cholinesterase inhibitory activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes. The antioxidant properties were evaluated using DPPH and ABTS methods. The total phenolic content (TPC) was measured by spectrometric assay, and compound identification was carried out by LC-MS/MS analysis. The results showed that the leaf and stem extracts of A. sericicarpus exerted significant inhibitory effects against AChE and BChE, as well as antioxidant activities. The stem ethanolic extract exhibited the highest potency against AChE and BChE with IC50 values of 5.81 and 11.46 µg/mL, respectively. The leaf and stem ethanolic extracts gave higher antioxidant activities and TPC compared to the water-based extracts. The LC-MS/MS analysis indicated the presence of phenolic compounds, such as flavones, flavonols, flavanones, prenylated chalcones, and xanthones in the extracts.

Tectona grandis has been reported for its composition of two 5α-reductase inhibitors, specifically (+)-eperua-8,13-dien-15-oic acid (1) and (+)-eperua-7,13-dien-15-oic acid (2). These compounds exhibit potential as markers, enabling the standardization of leaf extracts during pre-formulation and formulation development. This study examined the effects of solvent system, pH, temperature and light on the stability of compounds 1 and 2 in the ethanolic extract and the ethanolic extract in PG with PEG-40 as a solubilizer. The remaining amount of the compounds at all test conditions was analyzed using HPLC. Compounds 1 and 2 exhibited greater stability in acidic solutions compared to neutral or basic environments. Their degradations were susceptible to light, particularly compound 1 of the ethanolic extract in PG. Both compounds in the solution form of the extract degraded rapidly at high temperature, whereas the present of PG minimized their degradation when the light was protected. According to the Q10 method, the calculated shelf-life of the extracts was 1.48 years. The extracts should be stored or prepared as a non-aqueous form or otherwise in a slightly acidic solution with appropriate light protection measures. The pre-formulation information obtained from this study holds significant value in informing the design and development of forthcoming anti-hair loss medications. HIGHLIGHTS GRAPHICAL ABSTRACT

The objectives of this study were (1) to investigate the effect of extracts from some plants in the families Nelumbonaceae and Nymphaeaceae on phosphodiesterase 5 (PDE5) and arginase, which have been used in erectile dysfunction treatment, and (2) to isolate and identify the compounds responsible for such activities. The characterization and quantitative analysis of flavonoid constituents in the active extracts were performed by HPLC. Thirty-seven ethanolic extracts from different parts of plants in the genus Nymphaea and Victoria of Nymphaeaceae and genus Nelumbo of Nelumbonaceae were screened for PDE5 and arginase inhibitory activities. The ethanolic extracts of the receptacles and pollens of Nelumbo nucifera Gaertn., petals of Nymphaea cyanea Roxb. ex G.Don, Nymphaea stellata Willd., and Victoria amazonica (Poepp.) Sowerby and the petals and receptacles of Nymphaea pubescens Willd. showed IC₅₀ values on PDE5 of less than 25 μg/mL while none of the extracts showed effects on arginase. The most active extract, N. pubescens petal extract, was fractionated to isolate and identify the PDE5 inhibitors. The results showed that six flavonoid constituents including quercetin 3’-O-β-xylopyranoside (1), quercetin 3-methyl ether 3’-O-β-xylopyranoside (2), quercetin (3), 3-O-methylquercetin (4), kaempferol (5) and 3-O-methylkaempferol (6) inhibited PDE5 with IC₅₀ values at the micromolar level.

Background. Cassia spectabilis is a flowering plant containing various metabolites that provide potential for pharmacological activities. The current study aimed to investigate the ethanolic and water extracts of C. spectabilis as cholinesterase inhibitor as one of the target treatments for Alzheimers disease. The chemical composition of the extracts was also studied to determine which components are responsible for the bioactivity. Methods. The cholinesterase inhibitory activity assay was carried out by the modified Ellmans method against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). LC-MS/MS analysis was carried out to investigate the chemical profiles of the extracts, followed by a molecular networking study by GNPS. Results. Both extracts showed inhibition against AChE and BChE in a dose-dependent manner, with the higher potency exhibited by the ethanolic extract with IC50 values of 7.88 and 3.78μg/mL. The chemical analysis and molecular networking study of the flower extracts revealed similarity between the ethanolic and water extracts. Piperidine alkaloids were identified in both extracts, while the sphingolipid compounds were found in the ethanolic extract. Conclusion. The water and ethanolic extracts of C. spectabilis flowers displayed potency for Alzheimers disease treatment. The presence of piperidine alkaloids in the extract may be responsible for the cholinesterase inhibitory activity. The higher potency of the ethanolic extract compared to the water extract is possibly due to the higher amount of piperidine alkaloids in the ethanolic extract. Further study is needed to quantify the concentration of alkaloids in the extracts.

Steroid 5α-reductase plays a crucial role in catalyzing the conversion of testosterone to dihydrotestosterone, which is involved in many androgen-dependent disorders. Leaf-hexane extract from Tectona grandis L.f. has shown promise as a 5α-reductase inhibitor. The objectives of this current study were to isolate and identify 5α-reductase inhibitors from T. grandis leaves and to use them as the bioactive markers for standardization of the extract. Three terpenoid compounds, (+)-eperua-8,13-dien-15-oic acid (1), (+)-eperua-7,13-dien-15-oic acid (2), and lupeol (3), were isolated and evaluated for 5α-reductase inhibitory activity. Compounds 1 and 2 exhibited potent 5α-reductase inhibitory activity, while 3 showed weak inhibitory activity. An HPLC method for the quantitative determination of the two potent inhibitors (1 and 2), applicable for quality control of T. grandis leaf extracts, was also developed. The ethanolic extract showed a significantly higher content of 1 and 2 than found in the hexane extract, suggesting that ethanol is a preferable extraction solvent. This study is the first reported isolation of 5α-reductase inhibitors (1 and 2) from T. grandis leaves. The extraction and quality control methods that are safe and useful for further development of T. grandis leaf extract as an active ingredient for hair loss treatment products are also reported.

Triple negative breast cancer is one of the most aggressive breast cancer type with abilities of early metastasis and chemoresistance. The tropical plant Citrus hystrix DC. has been reported to promote many biological activities including anticancer. However, the effect of C. hystrix against triple negative breast cancer has not yet been identified. This study aimed to evaluate the anticancer properties of C. hystrix leaf extract and its bioactive constituents citronellol and citronellal against the triple negative breast cancer MDA-MB-231 cell line. C. hystrix leaves were powdered and sequentially macerated. The in vitro anticancer effects of C. hystrix leaf extracts, and its bioactive constituents (citronellol and citronellal) were evaluated against MDA-MB-231 cell line using cytotoxic MTT assay, cell proliferation, wound scratch migration, colony formation, cell cycle, apoptosis assay, Hoechst staining, RT-qPCR, and Western blot analysis. Results showed that crude hexane extract, citronellol, and citronellal significantly reduced cell proliferation, colony formation, and cell migration by inducing cell cycle arrest, while also inducing apoptosis in MDA-MB-231 cells through inhibition of anti-apoptotic Bcl-2 expression, leading to activation of the caspase-3-dependent pathway. This study is the first report to demonstrate the effect of C. hystrix, citronellol, and citronellal against triple negative breast cancer MDA-MB-231 cells.