Pinumin A,a Stilbenoid from the Roots of Pinus massoniana with Nitric Oxide Inhibitory Effect

Abstract

Objectives

This work aimed to isolate and characterize the chemical constituents of the roots of Pinus massoniana Lamb. and to evaluate their anti-inflammatory activity by assessing the inhibition of nitric oxide release in LPS-induced macrophages.

Methods

The compounds were isolated after extensive separation and purification. Structural elucidation of the new compound was performed using a combination of NMR spectroscopy, experimental circular dichroism (CD) spectroscopy, and computational CD analysis, whereas known compounds were identified by comparing their spectroscopic data with those reported in the literature. All compounds were evaluated for nitric oxide inhibition using the Griess assay.

Results

One new compound, namely pinumin A (1), was isolated alongside thirteen known compounds: cedrusinin (2), (7S,8R)-dihydrodehydrodiconiferyl alcohol (3), (2R,3R)-2,3-bis(4-hydroxy-3-methoxybenzyl)butyrolactone (4), nortrachelogenin (5), nectandrin B (6), machilin I (7), (+)-tsugacetal (8), (‒)-pinoresinol (9), pinostilbene (10), pinosylvin (11), resveratrol (12), (S)-8-hydroxycarvotanacetone (13) and isosclerone (14). Compounds 1, 5, and 8 demonstrated inhibition of LPS-induced NO production, with IC₅₀ values of 36.9 μM, 47.9 μM, and 39.3 μM, respectively.

Conclusion

One new compound, namely pinumin A (1) and thirteen known compounds were isolated from the roots of Pinus massoniana. Compounds 1, 5, and 8 demonstrated inhibition of LPS-induced NO production, with IC₅₀ values of 36.9 μM, 47.9 μM, and 39.3 μM, respectively. These results provide a reference for the subsequent discovery of anti-inflammatory active substances.

Keywords

pinaceae lignans nitric oxide release stilbenoid anti-inflammatory activity

Introduction

Chronic inflammation plays a role in the pathogenesis of a wide range of chronic diseases, such as Alzheimer's disease, cardiovascular disorders, and so on.¹ This persistent, low-grade inflammation promotes tissue damage and disease progression through the sustained release of harmful mediators like pro-inflammatory reactive oxygen species, cytokines and chemokines. Critically, its pathological overproduction of nitric oxide (NO) during chronic inflammation exerts significant detrimental effects. This pathological level of NO can cause DNA damage, inhibit protein function, induce mitochondrial dysfunction, and reactions with other reactive molecules to form more destructive reactive nitrogen species (RNS), such as peroxynitrite anions (ONOO⁻). These mechanisms exacerbate oxidative stress and tissue injury, profoundly participating in the pathophysiology of chronic diseases.^2,3

Natural products are important for drug research and development because of their complex and diverse molecular structures.⁴ Pinus massoniana Lamb. (Pinaceae) is an important tree species in China, and its different parts are rich in a variety of bioactive constituents. Its nodes,⁵ root bark,⁶ branches,^7–10 needles,^11,12 and pollen^13,14 were phytochemically investigated. These works report diterpenoids,^5–7 triterpenoids,⁸ procyanidins,^9,10 flavonoids,¹¹ polysaccharides,¹² phenylmethanoids,¹⁵ and phenylpropanoids¹³ as main metabolites.

The root of P. massoniana, commonly known as “Song geng” in Mandarin is native to numerous provinces across southern China. In folk medicinal material, it is used as a rheumatid-relieving, for detoxification and to treat falls and injuries. Furthermore, the roots of P. massoniana serve as the natural and essential substrate for Poria cocos, a key medicinal and edible fungus. The development of the Poria sclerotium is entirely dependent on nutrients absorbed from the pine root. This sclerotium is then directly processed into various traditional Chinese functional foods, such as Fuling biscuits and soups. Therefore, the pine root demonstrates its dietary importance indirectly but fundamentally by being the primary material foundation for this widely consumed food-medicine.¹⁴

Diterpenoids from P. massoniana significantly suppressed lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) overproduction in macrophages.⁷ Additionally, pine pollen demonstrated multi-target anti-inflammatory effects by directly inhibiting the production of inflammatory mediators and modulating key signaling pathways to regulate inflammation-related gene expression.¹⁶ Therefore, a systematic investigation into the chemical constituents and bioactivities of P. massoniana might be of value for identifying novel NO inhibitors.⁶ The roots were phytochemically investigated once, with the study focusing on the root bark.⁶ Herein, we report the first comprehensive investigation of the entire root. In this work, a total of fourteen compounds were isolated from the ethyl acetate extract of the roots of P. massoniana. They include one new (1) and thirteen known compounds. Herein, we detail the purification, structure analysis, and anti‒inflammatory evaluation of the compounds.

Materials and Methods

General Experimental Procedures

The key data for structure elucidation were collected on the following instruments: optical rotations on a Horiba SEPA-300 polarimeter; UV, IR (KBr pellets), and NMR (¹H, ¹³C, 2D) spectra were acquired on a Shimadzu UV-2401PC spectrophotometer, a Bruker Tensor 27 spectrometer, and a Bruker DRX-500 spectrometer, respectively. High-resolution mass spectrometry was conducted using an Applied Biosystems Mariner 5140 instrument.

To isolate and purify the constituents, TLC and column chromatography were conducted on pre-coated silica gel GF254 plates and silica gel (200‒300 mesh; Qingdao Marine Chemical), respectively, while Sephadex LH-20 was also employed. Final purification was performed using a Sep Sciences MH-LC52 medium-high pressure preparative liquid chromatography system (Sep Sciences, Beijing, China, Thermo column, 5 μm, 10 × 250 mm). All solvents met or exceeded the purity of analytical grade (≥95%).

Plant Material

The roots of Pinus massoniana Lamb. were collected from Mountain Dong ‘an County, Hunan Province, China, on July 15, 2024. The species was identified by Prof. X. J. Zhou from a specimen of the whole plant. A corresponding voucher sample (ZXJ-0157) is kept in the research laboratory at Hunan University of Chinese Medicine for future reference.

Extraction and Isolation

Extraction of the plant material (6.0 kg dried roots of P. massoniana) was performed using 60% aqueous ethanolic solvent (2 × 24 L). The next step was to concentrate the extract under reduced pressure. The material was then successively extracted with petroleum ether and EtOAc (6 × 4.0 L for each solvent). The petroleum ether extract showed no significant inhibitory activity against LPS-induced NO production in preliminary screening and was therefore not subjected to further phytochemical investigation.

Column chromatographic separation of the EtOAc extract (78.5 g) over silica gel employing a gradient of petroleum ether/EtOAc (15:1, 5:1, 3:1, 1:1, 1:2, 1:4) gave eight fractions (Frs. 1-8). Four subfractions (Frs. 4-1 to 4-4) were obtained from Fr. 4 (3.43 g) by Sephadex LH-20 column chromatography (MeOH). Frs. 4-4 (660 mg) was separated using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 60% aqueous MeOH to give compound 10 as a colorless oil (2.1 mg; t_R 24.49 min) and compound 11 as a yellow amorphous solid (15.2 mg; t_R 30.03 min). Fractionation of Fr. 4-1 (2.15 g) on a silica gel column using a stepwise petroleum ether/EtOAc gradient (8:1, 4:1, 2:1, 1:1, 1:2, 1:4) gave five subfractions (Frs. 4-1-1-4-1-5). Frs. 4-1-2 (820 mg) was further fractionated by Sephadex LH-20 column (MeOH), giving five subfractions (Frs. 4-1-2-1-4-1-2-5). Frs. 4-1-2-3 (280 mg) was separated using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 20% aqueous acetonitrile to give compound 13 as a colorless oil (3.9 mg; t_R 28.88 min). Frs. 4-1-2-4 (140 mg) was purified using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 25% aqueous acetonitrile to give compound 14 as a yellow amorphous solid (2.1 mg; t_R 28.74 min). Frs. 4-1-4 (630 mg) was further fractionated over Sephadex LH-20 column (MeOH) and then purified using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 40% aqueous acetonitrile to give compound 1 as a white amorphous solid (3.0 mg; t_R 22.12 min), compound 8 as a white amorphous solid (2.0 mg; t_R 28.32 min), compound 6 as a colorless oil (2.0 mg; t_R 51.33 min) and compound 7 as a colorless oil (4.0 mg; t_R 58.55 min). Separation of Fr. 5 (3.7 g) by RP-C18 chromatography, using a MeOH/H₂O gradient (30:70 to 100:0), afforded ten subfractions (Frs. 5-1-5-10). Frs. 5-1 (870 mg) was further fractionated over Sephadex LH-20 (MeOH), yielding four subfractions (Frs. 5-1-1－5-1-4). Frs. 5-1-2 (230 mg) was purified using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 45% aqueous MeOH to give compound 5 as a white amorphous solid (3.1 mg; t_R 25.53 min). Frs. 5-1-4 (43.6 mg) was also purified using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 60% aqueous MeOH to give compound 12 as a colorless oil (2.1 mg; t_R 34.72 min). Frs. 5-2 (69.7 mg) was fractionated over Sephadex LH-20 (MeOH) and then purified using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 45% aqueous acetonitrile to yield compound 9 as a white amorphous solid (7.7 mg; t_R 32.45 min) and compound 4 as a white amorphous solid (8.5 mg; t_R 37.72 min). Frs. 6 (3.96 g) was fractionated over Sephadex LH-20 (MeOH) to give four subfractions (Frs. 6-1－6-4). Fr. 6-2 (2.84 g) was separated over silica gel using a petroleum ether/EtOAc gradient (6:1, 4:1, 2:1, 1:1, 1:2, 1:4) to give five subfractions (Frs. 6-2-1－6-2-5). Frs. 6-2-3 (430 mg) was separated into six subfractions (Frs. 6-2-3-1－6-2-3-6) using Sephadex LH-20 (MeOH). Frs. 6-2-3-4 (80.0 mg) was purified using a medium and high-pressure preparation liquid phase system (MH-LC), eluting with 50% aqueous MeOH to give compound 3 as a yellow amorphous solid (18.4 mg; t_R 16.53 min) and compound 2 as a white amorphous solid (4.0 mg; t_R 24.13 min).

No other secondary metabolites were found in sufficient amount or purity to be characterized in the other subfractions of the EtOAc extract.

Pinumin A (1): white solid; mp 222 °C. [α]²⁵_D ‒7.9 (c 0.20, MeOH); IR (KBr) ν_max 3429, 2923, 2853, 1738, 1652, 1601, 1457, 1384, 1281, 1157, 1061, 843, 754, 701 cm^‒1; ¹H NMR and ¹³C NMR data, see Table 1 ; HRESIMS m/z 267.1001 [M + Na]⁺ (calcd for C₁₅H₁₆O₃Na⁺, 267.0997).

Table 1.

¹H (600 MHz) and ¹³C NMR (151 MHz) Spectroscopic Data of 1 in CD₃OD.

Position	δ_C, Type	δ_H (J in Hz)
1	140.5, C	–
2	106.1, CH	6.16, br s
3	160.6, C	–
4	98.9, CH	6.18, t (2.2)
5	157.8, C	–
6	108.8, CH	6.22, br s
7	45.7, CH₂	2.96, dd (13.4, 7.3) 2.85, dd (13.4, 6.4)
8	75.1, CH	4.82, t (6.9)
3-OMe	54.1, CH₃	3.67, s
1′	144.4, C	–
2′	125.9, CH	7.30, overlapped
3′	127.7, CH	7.30, overlapped
4′	126.8, CH	7.24, m
5′	127.7, CH	7.30, overlapped
6′	125.9, CH	7.30, overlapped

ECD Calculation

We used Sybyl-X 2.0 to sample the conformations and used the MMFF94S force field (energy cutoff value: 5 kcal/mol) to obtain nine low-energy conformations. These structures were optimized and frequencies calculated in ORCA 5.0.1. using the B3LYP-D3(BJ)/6-31G* method and CPCM (methanol solvent)¹⁷ confirming that they are all local minima (no imaginary frequencies). Then, TD-DFT calculations were performed using the PBE0/def2-TZVP method in CPCM (methanol solvent)¹⁸ and the excitation energies, oscillator strengths, and rotatory strengths for the sixty lowest-energy-excited states were calculated. The ECD spectrum was simulated using a Gaussian function (σ = 0.30).¹⁹ The Gibbs free energy of the conformation was calculated from the thermal correction of B3LYP-D3(BJ)/6-31G and the electron energy of wB97M-V/def2-TZVP. Finally, the Boltzmann-weighed spectrum was simulated based on the relative ΔG values and compared with experimental data applied to the configurational assignment of the chiral center.

Biological Assays

A density of 10⁵ cells/mL was used to seed RAW 264.7 cells (Cangsha, Abiowell, China) in 96-well plates. Then, the cells were subjected to a 24-h culture period at 37 °C within a 5% CO₂ incubator. To evaluate anti-inflammatory activity, cells were first primed with the compounds for 1 h after the initial 24 h culture, followed by LPS (1 μg/mL) challenge for 24 h. The inhibitory effect on NO generation was determined by measuring absorbance at 540 nm using the Griess reagent. A series of standard solutions were prepared with NaNO₂ as the standard to draw the standard curve. Based on the measured absorbance values, the corresponding nitrite concentration was determined, from which the inhibition rate of NO release was calculated. The positive control was dexamethasone, while cell viability was determined by the CCK-8 method. All experiments were independently performed five times, and the data are expressed as the mean ± SD. Statistical significance was determined using a student's t-test, with a p-value of ≤ 0.05 considered significant.

Results and Discussion

Compound 1 was isolated as a white amorphous solid. Its molecular formula (C₁₅H₁₆O₃) was deduced from the sodium adduct at m/z 267.1001 in the HRESIMS spectrogram, with a degree of unsaturation of 8. The IR spectrum implied the presence of hydroxy groups (3429 cm^‒1) and benzene rings (1652, 1600 and 1560 cm^‒1). The ¹H NMR spectrum showed three aromatic proton signals at δ_H 6.22 (1H, s), 6.18 (1H, t, J = 2.2 Hz), and 6.16 (1H, s), suggesting the presence of a 13,5-trisubstituted benzene ring ( Table 1 ). The ¹³C and HSQC spectroscopic data indicated the presence of fifteen aromatic carbons. They include two oxygenated aromatic quaternary carbons (δ_C 160.6 and 157.8), two aromatic quaternary carbons (δ_C 144.4 and 140.5), eight aromatic methine carbons (δ_C 127.7, 126.8, 125.5, 125.5, 108.8, 106.1 and 98.9), one oxygenated methine carbon (δ_C 75.1), and one methoxy carbon (δ_C 54.1). The planar structure of 1 was elucidated using key COSY correlations of H-3′/H-4′/H-5′ and H₂-7/H-8, and key HMBC correlations of H-2 to C-4 and C-7; H-6 to C-2, C-4 and C-7; H-8 to C-1, C-1′, C-2′ and C-6′; and of 3-OMe to C-3 (Figure 1). The spectroscopic data suggested that compound 1 closely resembled 3-O-methyldihydropinosylvin,²⁰ except for H-8, which was substituted by a hydroxy group. This conclusion was supported by the ¹H and ¹³C NMR spectroscopic data of 1 (Table 1). Comparison of the calculated ECD spectrum for the (8S) enantiomer with the experimental ECD spectrum of 1 showed a good match ( Figure 2 ), thereby confirming the 8S absolute configuration of compound 1. Compound 1 was thus named pinumin A.

Figure 1.

Key COSY (bold) and HMBC (arrows) correlations for compound 1.

Figure 2.

Experimental and calculated ECD spectra of 1.

The known compounds were identified as cedrusinin (2),²¹ (7S,8R)-dihydrodehydrodiconiferyl alcohol (3),²² (2R,3R)-2,3-bis(4-hydroxy-3-methoxybenzyl) butyrolactone (4),²³ nortrachelogenin (5),²⁴ nectandrin B (6),²⁵ machilin I (7),²⁵ (+)-tsugacetal (8),²⁶ (‒)-pinoresinol (9),²⁷ pinostilbene (10),²⁸ pinosylvin (11),²⁸ resveratrol (12),²⁹ (S)-8-hydroxycarvotanacetone (13)³⁰ and isosclerone (14).³¹ fig 3.

Figure 3.

Skeletal (shorthand) structures of the isolated compounds (1-14).

As the roots of P. massoniana are used in the treatment of conditions related to chronic inflammation such as rheumatic swelling and pain, the isolated compounds were tested for LPS-induced NO production in RAW 264.7 cells using the Griess method. Dexamethasone was used as a positive control. At a concentration of 100 μM, compounds 1‒14 inhibited LPS-induced NO production in RAW264.7 cells. Compounds 1, 5, and 8 demonstrated the most potent activity, with inhibition rates exceeding 50% (see Supplementary Table S1). Consequently, the IC₅₀ values were determined, with a parallel assessment of cell viability via the CCK-8 method. The results revealed that Compounds 1, 5, and 8 moderate demonstrated inhibition of LPS-induced NO production, with respective IC₅₀ values of 36.9, 47.9 and 39.3 μM. The positive control, dexamethasone, exhibited potent activity with an IC₅₀ value of 117.1 nM.

However, this study has several limitations. The anti-inflammatory activity was evaluated only in an LPS-induced RAW 264.7 macrophage model, focusing on NO suppression without elucidating the underlying mechanism. Therefore, a deeper understanding of their mechanisms, selectivity, and in vivo efficacy is required before their therapeutic potential can be pursued, necessitating future work on comprehensive pharmacokinetic profiling, toxicity assessment, and structure optimization. (Table 2) (Figure 4).

Figure 4.

Inhibitory curves of compounds 1, 5 and 8 on LPS-induced NO production in RAW 264.7 cells.

Table 2.

Inhibition of NO Production in LPS-Induced RAW 264.7 Cells by Compounds 1, 5, and 8.

Compounds	Concentration (μM)	NO Production (μM)	Inhibition Rate (%)	IC₅₀ [95% CI] (SD)
Control		2.6 ± 0.1
LPS		65.4 ± 1.5
	1	48.4 ± 1.6	27.0 ± 2.6
	25	41.1 ± 1.4	38.7 ± 2.2
1	50	25.1 ± 0.7	61.4 ± 1.4	39.6 μM[36.4–42.7](2.6 μM)
	75	18.9 ± 1.1	74.1 ± 1.7
	100	14.4 ± 0.7	81.2 ± 1.2
	1	51.5 ± 1.6	22.1 ± 2.6
	25	42.8 ± 0.9	35.9 ± 1.4
5	50	31.5 ± 1.2	53.9 ± 1.9	36.9 μM[35.1–38.7](2.1 μM)
	75	24.4 ± 0.8	65.2 ± 1.3
	100	16.3 ± 0.7	78.1 ± 1.2
	1	48.3 ± 1.4	28.0 ± 2.3
	25	37.9 ± 1.7	43.8 ± 2.8
8	50	29.3 ± 1.5	57.3 ± 2.3	47.9 μM[47.0–48.7](1.9 μM)
	75	22.3 ± 1.1	68.5 ± 1.7
	100	15.8 ± 0.9	78.8 ± 1.5

Data are mean IC₅₀ [95% CI] (SD of IC₅₀ values, n = 5), derived from four-parameter logistic (4PL) curve fits of five independent replicates, the overall mean and CI were calculated using a t-distribution model.

Conclusion

In this study, one new compound, namely pinumin A, and thirteen known compounds were isolated from the roots of P. massoniana. Compounds 1, 5, and 8 exhibited weak NO release inhibition ability. These results lend scientific support to the traditional use of the roots of P. massoniana for treating rheumatism, detoxification, and traumatic injuries, ailments often involving inflammatory pathways mediated by excessive NO release. The discovery of pinumin A enriches the structural diversity of phenolic compounds, particularly stilbenoids, from Pinus species, and highlights the chemical potential of not fully explored plant parts in an otherwise well-studied tree. Furthermore, these results provide a reference for the subsequent discovery of anti-inflammatory active substances.³²

Supplemental Material

sj-doc-1-npx-10.1177_1934578X261433098 - Supplemental material for Pinumin A, a Stilbenoid from the Roots of Pinus massoniana with Nitric Oxide Inhibitory Effect

Supplemental material, sj-doc-1-npx-10.1177_1934578X261433098 for Pinumin A, a Stilbenoid from the Roots of Pinus massoniana with Nitric Oxide Inhibitory Effect by Hongyu Jiang, Yuxin Peng, Guangyun Su, Xining Sun, Xuejing Li and Xiaojiang Zhou in Natural Product Communications

Footnotes

Acknowledgments

This work was supported by Science and Medicine Joint Fund of Hunan Provincial National Natural Science Foundation, China (No. 2024JJ8160).

ORCID iD

Xiaojiang Zhou

Ethical Statement

This study did not involve any human participants, human data, animal experiments, or animal tissues. The research solely focused on the phytochemical investigation of plant material (Pinus massoniana). Therefore, ethical approval from an institutional review board or ethics committee was not required for this work.

Funding

The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by the Medicine Joint Fund of Hunan Provincial National Natural Science Foundation, (grant number 2024JJ8160).

Declaration of Conflicting Interests

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Supplemental Material

Supplementary data (NMR spectra and HRESIMS) for compound 1 are available in the online version.

References

Ferguson

. Chronic inflammation and mutagenesis. Mutat Res. 2010;690(1-2):3-11. doi: 10.1016/j.mrfmmm.2010.03.007

Nathan

. Points of control in inflammation. Nature. 2002;420(6917):846-852. doi:10.1038/nature01320

Förstermann

Sessa

. Nitric oxide synthases: regulation and function. Eur Heart J. 2012;33(7):829-837d. doi:10.1093/eurheartj/ehr304

Hong

. Role of natural product diversity in chemical biology. Curr Opin Chem Biol. 2011;15(3):350-354. doi: 10.1016/j.cbpa.2011.03.004

Chen

Zhang

, et al. Hepatoprotective diterpenes from the nodes of Pinus massoniana. J Asian Nat Prod Res. 2025;27(7):995-1004. doi:10.1080/10286020.2025.2506527

Ding

Zhang

, et al. Diterpenoids from the root bark of Pinus massoniana and evaluation of their phosphodiesterase type 4D inhibitory activity. J Nat Prod. 2020;83(4):1229-1237. doi: 10.1021/acs.jnatprod.9b01269

Zhang

, et al. Two new abietane diterpenoids from Pinus massoniana. J Asian Nat Prod Res. 2026;28(3):405-412. doi:10.1080/10286020.2025.2509768

Gao

, et al. Isolation and characterization of triterpenoids from the stem barks of Pinus massoniana. Holzforschung. 2017;71(9):697-703. doi:10.1515/hf-2016-0228

Zhou

Alania

Reis

, et al. Seco B-type oligomers from Pinus massoniana expand the procyanidin chemical space and exhibit dental bioactivity. J Nat Prod. 2022;85(12):2753-2768. doi: 10.1021/acs.jnatprod.2c00664

10.

Zhou

Alania

Reis

, et al. Rare A-type, spiro-type, and highly oligomeric proanthocyanidins from Pinus massoniana. Org Lett. 2020;22(14):5304-5308. doi: 10.1021/acs.orglett.0c01439

11.

Shen

Theander

. Flavonoid glycosides from needles of Pinus massoniana. Phytochemistry. 1985;24(1):155-158. doi:10.1016/S0031-9422(00)80826-2

12.

Chu

Yang

Lin

, et al. Chemical composition, antioxidant activities of polysaccharide from pine needle (Pinus massoniana) and hypolipidemic effect in high-fat diet-induced mice. Int J Biol Macromol. 2019;125:445-452. doi: 10.1016/j.ijbiomac.2018.12.082

13.

Zhang

Gong

Yang

, et al. Antioxidant constituents from Pinus massoniana (Pinaceae). Plant Divers Resour. 2013;35(5):615-622. doi:10.7677/ynzwyj201312104

14.

Gao

Liu

Sun

Cui

. Species diversity, taxonomic classification and ecological habits of polypore fungi in China. Mycology. 2024;16(2):419-544. doi:10.1080/21501203.2024.2384567

15.

Sun

Zheng

Zhong

Liu

. Chemical constituents of pollen from Pinus massoniana. Zhong Cao Yao. 2010;41(4):530-532.

16.

Lee

Kim

Choi

. Antioxidant and antiinflammatory activity of pine pollen extract in vitro. Phytother Res. 2009;23(1):41-48. doi:10.1002/ptr.2525

17.

Neese

Wennmohs

Becker

Riplinger

. The ORCA quantum chemistry program package. J Chem Phys. 2020;152(22):224108. doi:10.1063/5.0004608

18.

Superchi

Scafato

Gorecki

Pescitelli

. Absolute configuration determination by quantum mechanical calculation of chiroptical Spectra: basics and applications to fungal metabolites. Curr Med Chem. 2018;25(2):287-320. doi:10.2174/0929867324666170310112009

19.

Bruhn

Schaumlöffel

Hemberger

Bringmann

. Specdis: quantifying the comparison of calculated and experimental electronic circular dichroism spectra. Chirality. 2013;25(4):243-249. doi:10.1002/chir.22138

20.

Hanawa

Yamada

Nakashima

. Phytoalexins from Pinus strobus bark infected with pinewood nematode, Bursaphelenchus xylophilus. Phytochemistry. 2001;57(2):223-228. doi:10.1016/s0031-9422(00)00514-8

21.

Sinkkonen

Karonen

Liimatainen

Pihlaja

. Lignans from the bark extract of Pinus sylvestris L. Magn Reson Chem. 2006;44(6):633-636. doi:10.1002/mrc.1780

22.

Kuang

Xia

Yang

Wang

Lü

. Lignan constituents from Chloranthus japonicus Sieb. Arch Pharm Res. 2009;32(3):329-334. doi:10.1007/s12272-009-1303-1

23.

Jin

Zhao

Nakamura

Akao

Kakiuchi

Hattori

. Isolation and characterization of a human intestinal bacterium, Eubacterium sp. ARC-2, capable of demethylating arctigenin, in the essential metabolic process to enterolactone. Biol Pharm Bull. 2007;30(5):904-911. doi:10.1248/bpb.30.904

24.

Tang

Peng

Wen

, et al. Chemical constituents of Artemisia frigida. Chem Nat Compd. 2022;58(4):735-737. doi:10.1007/s10600-022-03780-0

25.

Filleur

Bail

Duroux

Simon

Chulia

. Antiproliferative, anti-aromatase, anti-17beta-HSD and antioxidant activities of lignans isolated from Myristica argentea. Planta Med. 2001;67(8):700-704. doi:10.1055/s-2001-18349

26.

Fang

Wei

Cheng

Wang

. (+)-Tsugacetal, a lignan acetal from Tsuga chinensis. Phytochemistry. 1985;24(6):1363-1365. doi:10.1016/S0031-9422(00)81134-6

27.

Schöettner

Reiner

Tayman

FSK

. (+)-Neoolivil from roots of Urtica dioica. Phytochemistry. 1998;46(6):1107-1109. doi:10.1016/S0031-9422(97)00401-9

28.

Kırmızıbekmez

Aru

Křoustková

, et al. Cytotoxic stilbenoids, hetero- and homodimers of homoisoflavonoids from Prospero autumnale. J Nat Prod. 2025;88(3):458-468. doi: 10.1021/acs.jnatprod.4c01263

29.

Cheng

Fang

Chen

Zhou

Yang

Liu

. Structure-activity relationship studies of resveratrol and its analogues by the reaction kinetics of low density lipoprotein peroxidation. Bioorg Chem. 2006;34(3):142-157. doi:10.1016/j.bioorg.2006.04.001

30.

Sousa

Leitão

Neves

Cavaleiro

Mendes

. Standardised comparison of limonene-derived monoterpenes identifies structural determinants of anti-inflammatory activity. Sci Rep. 2020. Published 2020 Apr 29;10(1):7199. doi:10.1038/s41598-020-64032-1

31.

Cimmino

Villegas-Fernández

Andolfi

Melck

Rubiales

Evidente

. Botrytone, a new naphthalenone pentaketide produced by Botrytis fabae, the causal agent of chocolate spot disease on Vicia faba. J Agric Food Chem. 2011;59(17):9201-9206. doi:10.1021/jf202089y

32.

Shimomura

Sashida

Oohara

. Lignans from Machilus thunbergii. Phytochemistry. 1987;26(5):1513-1515. doi:10.1016/S0031-9422(00)81847-6

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

6.35 MB