Use of urushiol derivatives in dry lacquer extract in preparation of drugs for resisting myocardial fibrosis

Abstract

The application provides application of urushiol derivatives in a lacquer extract in preparation of anti-myocardial fibrosis drugs. The urushiol derivative is shown as a structural general formula I. In the formula I, R1 represents H or -OH, R2 represents a straight-chain alkyl of C 10 -C 20 , or a straight-chain alkyl of C 10 -C 20 containing at least one substituent of a double bond, C=O, COOH, OH, or -Q-R4, wherein Q represents a straight-chain alkyl of C 10 , and R4 represents a substituted or unsubstituted benzene ring, or -QCO-R4, wherein Q represents a straight-chain alkyl of C 10 , and R4 represents a substituted or unsubstituted benzene ring, or -QCOO-R5, wherein Q represents a straight-chain alkyl of C 10 , and R5 represents C1-C6 alkyl; and R3 represents at least one of H, -OH, HCO-, CH3CO-, CH3CH2CO-. The results of a pharmacological activity experiment show that the compound has better inhibition activity on myocardial fibrosis.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to application of a laccaic acid derivative in an extract of dry lacquer in preparation of a drug for resisting myocardial fibrosis. BACKGROUND

[0002] Toxicodendron vernicifluum (Stokes) F.A.Barkl., Rhus verniciflua Stokes is a deciduous tree of Toxicodendron of Anacardiaceae. There are more than 20 species of Toxicodendron, mainly distributed in eastern Asia and North America to Central America, and also distributed in Korea, India and Japan. There are 15 species of Toxicodendron in China, which are important and unique economic tree species. The lacquer tree produces the highest amount of lacquer, such as Sichuan Chengkou lacquer in central south China, Maoba lacquer in Hubei, Dafang lacquer in Guizhou, Zhenxiong lacquer in Yunnan, An Kang lacquer (Niuwang lacquer) in Shaanxi, and “Yan lacquer” in east China, which are the best Chinese lacquer.

[0003] Dry lacquer (Toxicoendri Resina) is the resin of T. vernicifluum of Anacardiaceae, which is warm in nature, pungent in taste, and slightly toxic. It enters the liver, spleen and stomach channels. It has the effects of breaking blood stasis, removing obstruction, removing accumulation and killing insects. It has been included in all versions of Chinese Pharmacopoeia. It has the effects of breaking blood stasis, removing obstruction, removing accumulation and killing insects. In the early 1960s, dry lacquer was introduced into Pingxiao tablets, a pure traditional Chinese medicine preparation, mainly for its effective component urushiol, which has the effects of breaking blood stasis, removing obstruction, removing accumulation and killing insects. It is a traditional Chinese medicine for promoting blood circulation to remove blood stasis and relieving pain and resolving mass, which has been used in clinical treatment of cancer since 1958 and has certain effect in the treatment of lung cancer, liver cancer, esophageal cancer and bone tumor. Dahuang Ejia Worm Pill, which is composed of 12 ingredients including dry lacquer, is also widely used in clinical practice, involving more than 40 diseases, and has the effects of promoting blood circulation to remove blood stasis, softening and resolving mass, and relieving middle and tonifying deficiency. The traditional Chinese medicine dry lacquer lacks systematic research on its chemical components, and the relationship between its traditional effect of promoting blood circulation to remove blood stasis and its structure is rarely studied. There are no specific qualitative and quantitative detection indexes in the quality standard of dry lacquer in the current version (2020 version) of Chinese Pharmacopoeia, which makes it difficult to effectively control the quality of dry lacquer and its decoction pieces, and brings hidden dangers to clinical safe medication. Moreover, the research on the chemical components of traditional Chinese medicine dry lacquer is not comprehensive, and the relationship between its traditional effect of promoting blood circulation to remove blood stasis and its structure is even less studied.

[0004] Myocardial fibrosis is a common pathological process of heart failure, hypertension, coronary heart disease and various myocardial diseases at a certain stage, which is manifested as proliferation of myocardial interstitial myocardial fibroblasts, deposition of extracellular matrix, cell death and vascular regeneration. Myocardial fibrosis can affect myocardial metabolism, leading to ventricular remodeling and finally causing heart failure.

[0005] There is no research on the active ingredient of dry lacquer for anti-myocardial fibrosis drugs. SUMMARY

[0006] The present application provides a laccaic acid derivative in dry lacquer extract and its application in the medical field.

[0007] The laccaic acid derivative in the dry lacquer extract has a general structure as shown in formula I:

[0008]

[0009] In formula I, R1 represents H or -OH,

[0010] R2 represents a straight chain alkyl group of C5-C 20 (preferably C 10 -C 20 ) straight chain alkyl group, or

[0011] C5-C 20 (preferably C 10 -C 20 ) straight chain alkyl group (if two or more substituents are contained, the two or more substituents can be the same or different), or

[0012] -Q-R4, wherein Q represents a straight chain alkyl group of C6-C 10 , and R4 represents a substituted or unsubstituted benzene ring, or

[0013] -QCO-R4, wherein Q represents a straight chain alkyl group of C6-C 10 , and R4 represents a substituted or unsubstituted benzene ring, or

[0014] -QCOO-R5, wherein Q represents a straight chain alkyl group of C6-C 10 , and R5 represents a C1-C6 alkyl group,

[0015] wherein the substituents on the substituted benzene ring are C1-C6 alkyl groups;

[0016] R3 represents at least one of H, -OH, HCO-, CH3CO-, CH3CH2CO-.

[0017] The laccaic acid derivative in the dry lacquer extract is any one of the following compounds:

[0018]

[0019] The application of the laccaic acid derivative shown in formula I above in the preparation of an anti-myocardial fibrosis drug also belongs to the protection scope of the present application.

[0020] The application also provides an anti-myocardial fibrosis drug containing the urushibara derivative shown in formula I.

[0021] The application constructs an in-vitro myocardial fibrosis model by stimulating neonatal SD rat myocardial fibroblasts with 20 ng / mL TGF-β1 for 48 h, and detects the relative activity of the cells by CCK-8, with tanshinone II A as a positive drug. The results show that the compounds 1*, 2*, 5*, 6*, 8*, 10*, 12*, 14*, 19*, 20*, 21*, 29, 30, 33, 37 have better myocardial fibrosis inhibition activity and show concentration dependence; the myocardial fibrosis inhibition activity of the compound 35 shows poor concentration dependence; the myocardial fibrosis inhibition activity of the compounds 9*, 16* and 18* is high at a low concentration of 10 μM. Figure 24 BRIEF DESCRIPTION OF DRAWINGS

[0022] The accompanying drawings are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the principles of the application. In the drawings:

[0023] Figure 1 The structure of the compound 1* in Example 1 of the application, the main 1 H- 1 H COSY and HMBC correlation diagrams;

[0024] Figure 2 The EI-MS main fragments of the compound 1* in Example 1 of the application;

[0025] Figure 3 The structure of the compound 2* in Example 1 of the application, the main 1 H- 1 H COSY and HMBC correlation diagrams;

[0026] Figure 4 The structure of the compound 3* in Example 1 of the application, the main 1 H- 1 H COSY and HMBC correlation diagrams;

[0027] Figure 5 The structure of the compound 5* in Example 1 of the application, the main 1 H- 1 H COSY and HMBC correlation diagrams;

[0028] Figure 6 The HREI-MS main fragments of the compound 5* in Example 1 of the application;

[0029] Figure 7 The structure of the compound 6* in Example 1 of the application, the main​1 H- 1 H COSY and HMBC correlation diagrams;

[0030] Figure 8 EI-MS main fragments of compound 6* in Example 1 of the present application;

[0031] Figure 9 structure of compound 8* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlation diagrams;

[0032] Figure 10 structure of compound 9* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlation diagrams;

[0033] Figure 11 structure of compound 10* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlation diagrams;

[0034] Figure 12 structure of compound 12* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlation diagrams;

[0035] Figure 13 structure of compound 14* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlation diagrams;

[0036] Figure 14 structure of compound 16* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlation diagrams;

[0037] Figure 15 HREI-MS and EI-MS main fragments of compound 16* in Example 1 of the present application;

[0038] Figure 16 structure of compound 18* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlation diagrams;

[0039] Figure 17 HREI-MS and EI-MS main fragments of compound 18* in Example 1 of the present application;

[0040] Figure 18 Structure of compound 19* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlations;

[0041] Figure 19 HREI-MS and EI-MS main fragments of compound 19* in Example 1 of the present application;

[0042] Figure 20 Structure of compound 20* in Example 1 of the present application, main 1 H- 1 H COSY and HMBC correlations;

[0043] Figure 21 HREI-MS and EI-MS main fragments of compound 20* in Example 1 of the present application;

[0044] Figure 22 Structure of compound 21* in Example 1 of the present application; main 1 H- 1 H COSY and HMBC correlations;

[0045] Figure 23 HREI-MS and EI-MS main fragments of compound 21* in Example 1 of the present application;

[0046] Figure 24 Activity screening of small molecules (20 μM) based on TGF-β induced fibrosis model in Example 2 of the present application (compared with blank group: ### P<0.001; compared with model group: *P<0.05, **P<0.01, ***P<0.001). DETAILED DESCRIPTION

[0047] The preferred embodiments of the present application will be described herein below with reference to the drawings; it should be understood that the preferred embodiments described herein are intended to explain and describe the present application, and are not intended to limit the present application.

[0048] Example 1

[0049] The dry lacquer T. vermicifluum 50.3 kg was refluxed with 8 times amount of 95% ethanol for 3 times, each time for 2 h, and the filtrate was combined and the solvent was recovered under reduced pressure to obtain a total extract 2.2 kg. The extract was suspended in water and extracted with petroleum ether, ethyl acetate, n-butanol and water for 3 times, respectively, and the solvent was recovered. Finally, the petroleum ether extract part was 1123 g, the ethyl acetate part was 780 g, the n-butanol part was 77 g and the water part was 185 g.

[0050] The petroleum ether extract and ethyl acetate extract of dry lacquer were analyzed by HPLC with DAD detection. The UV absorption of the laccaic acid (λ max 210, 280 nm) and terpenoids (λ max 210 nm) were found. The petroleum ether extract and ethyl acetate extract of dry lacquer were analyzed by TLC. Most of the spots showed dark spots under UV light at 254 nm, and a few of the spots showed blue or sky blue fluorescence under UV light at 365 nm, indicating that the dry lacquer mainly contained laccaic acid and terpenoids, and a small amount of flavonoids. The 95% ethanol extract of dry lacquer was analyzed by LC-MS / MS, and it was found that the laccaic acid mainly produced characteristic fragments of m / z 122 and 106. Therefore, the separation strategy of TLC, HPLC and LC-MS / MS was adopted for the laccaic acid and terpenoids in dry lacquer.

[0051] The petroleum ether extract sample (1123 g) was mixed with silica gel, and then subjected to column chromatography on silica gel (100-200 mesh) for rough separation. The elution system was determined by pre-experiment to be nine gradient systems of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v), and each gradient elution was 3 column volumes. The chromatography was terminated by TLC detection, the flow fractions were combined, and the solvent was recovered, finally obtaining six sub-flow fractions (A-F).

[0052] Flow fraction B was subjected to column chromatography on silica gel under reduced pressure for further separation, and the elution system was nine gradient systems of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v). Four sub-flow fractions B1-B4 were finally obtained by combining the flow fractions according to their TLC behaviors. Flow fractions B1-B2 were observed as oily liquids, which were speculated to be low-polarity aliphatic compounds, so they were not processed temporarily. The separation work started from flow fraction B3 which was slightly more polar. B3 was separated by ODS reverse-phase column chromatography, and was eluted with methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in gradient, and the same components were combined to obtain flow fractions B3-a-B3-h. B3-g was purified by semi-preparative liquid chromatography (acetonitrile-water, 90:10, 3 mL / min) to obtain compounds 19* (8 mg), 20* (8.9 mg) and 21* (7.3 mg). B6-h was purified by semi-preparative liquid chromatography (acetonitrile-water, 95:5, 3 mL / min) to obtain compounds 22* (7.1 mg), 23* (8.5 mg) and 24* (9.2 mg).

[0053] Fractions C were re-fractionated by column chromatography on silica gel under reduced pressure using nine gradient systems of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) and combined on the basis of their thin layer behavior to yield nine sub-fractions C1-C9. C6 was first separated by column chromatography on Sephadex LH-20 gel (dichloromethane-methanol, 1:1) to yield five sub-fractions C6-a-C6-e: C6-b was purified by semi-preparative liquid chromatography on ODS (acetonitrile-water, 60:40, 3 mL / min) to yield compound 13 (8.6 mg); C6-c was purified by semi-preparative liquid chromatography on ODS (acetonitrile-water, 65:35, 3 mL / min) to yield compound 34 (7.8 mg), 36 (10.9 mg); C6-d was purified by semi-preparative liquid chromatography on ODS (acetonitrile-water, 70:30, 3 mL / min) to yield compound 37 (9.2 mg). Fractions D were separated by column chromatography on ODS using a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) and combined on the basis of their thin layer behavior to yield six sub-fractions D1-D6. D4 was re-fractionated by column chromatography on silica gel under reduced pressure using nine gradient systems of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) and combined on the basis of their thin layer behavior to yield three sub-fractions D4-a-D4-c. D4-a was purified by semi-preparative liquid chromatography on ODS (acetonitrile-water, 66:34, 3 mL / min) to yield compound 26 (9.5 mg), 27 (10.2 mg), 28 (8.7 mg), 29 (6.8 mg), 30 (9.2 mg). D4-c was purified by semi-preparative liquid chromatography on ODS (acetonitrile-water, 68:32, 3 mL / min) to yield compound 31 (7.7 mg), 32 (9.8 mg), 33 (8.4 mg). D5 was re-fractionated by column chromatography on silica gel under reduced pressure using nine gradient systems of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) and combined on the basis of their thin layer behavior to yield three sub-fractions D5-a-D5-c. D5-b was separated by column chromatography on ODS using a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) and combined to yield three fractions D5-b1-D5-b3. D5-b2 was purified by semi-preparative liquid chromatography on ODS (acetonitrile-water, 74:26 3 mL / min) to yield compound 16* (9.6 mg).D5-c was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) sequentially, and the fractions with the same component were combined to give six fractions D5-c1-D5-c6. D5-c6 was purified by semi-preparative HPLC (acetonitrile-water, 80:20, 3 mL / min) to give compound 1* (7.5 mg).

[0054] Fraction E was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) sequentially, and the fractions with the same component were combined to give seven sub-fractions El-E7. E6 was further separated by silica gel column chromatography under reduced pressure using a gradient of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) sequentially, and the fractions with the same component were combined to give three sub-fractions E6-a-E6-c. E6-b was purified by semi-preparative HPLC (acetonitrile-water, 66:34, 3 mL / min) to give compound 9* (8.9 mg) and 25 (10.3 mg). The ethyl acetate fraction (780 g) was first separated by silica gel column chromatography (100-200 mesh) using a gradient of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) sequentially, and the fractions with the same component were combined to give six sub-fractions (A-F).

[0055] Fraction B was further separated by silica gel column chromatography under reduced pressure using a gradient of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) sequentially, and the fractions with the same component were combined to give four sub-fractions Bl-B4. B4 was first separated by Sephadex LH-20 gel column chromatography (dichloromethane-methanol, 1:1) to give two sub-fractions B4-a-B4-b. B4-a was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) sequentially, and the fractions with the same component were combined to give five fractions B4-a1-B2-a5. B4-a4 was purified by semi-preparative HPLC (acetonitrile-water, 55:45, 3 mL / min) to give compound 35 (10.9 mg).

[0056] Fractions C were separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) and pooling fractions according to their thin layer behavior to give five sub-fractions C1-C5. C3 was first separated by Sephadex LH-20 gel column chromatography (dichloromethane-methanol, 1:1) to give two sub-fractions C3-a-C3-b. C3-b was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) and pooling fractions according to their thin layer behavior to give three fractions C3-b1-C3-b3. C3-b2 was purified by semi-preparative HPLC (acetonitrile-water, 55:45, 3 mL / min) to give compound 15* (6.9 mg). Fraction C4 was further separated by reduced pressure silica gel column chromatography eluting with a gradient of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) and pooling fractions according to their thin layer behavior to give three sub-fractions C4-a-C4-c. C4-a was purified by semi-preparative HPLC (acetonitrile-water, 55:45, 3 mL / min) to give compound 14* (9.5 mg). C4-c was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) and pooling fractions according to their thin layer behavior to give four fractions C4-c1-C4-c4. C4-c3 was purified by semi-preparative HPLC (acetonitrile-water, 60:40, 3 mL / min) to give compound 8* (10.2 mg), 18* (7.3 mg). Fraction C5 was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) and pooling fractions according to their thin layer behavior to give three fractions C5-a-C4-c. C5-c was purified by semi-preparative HPLC (acetonitrile-water, 65:35, 3 mL / min) to give compound 10* (11.2 mg), 11* (8.9 mg).

[0057] Fractions D were separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in order and pooling fractions according to their thin layer behavior, resulting in five sub-fractions D1-D5. D4 was further fractionated by silica gel column under reduced pressure using nine gradient systems of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) in order and pooling fractions according to their thin layer behavior, resulting in three sub-fractions D4-a-D4-c. D4-b was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in order and pooling fractions according to their thin layer behavior, resulting in three fractions D4-b1-D4-b3. D4-b2 was purified by semi-preparative liquid chromatography (acetonitrile-water, 65:35, 3 mL / min) to give compound 4* (7.6 mg). D5 was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in order and pooling fractions according to their thin layer behavior, resulting in four fractions D6-a-D6-d. D6-b was purified by semi-preparative liquid chromatography (acetonitrile-water, 68:32, 3 mL / min) to give compounds 12* (11.3 mg), 17* (7.3 mg).

[0058] Fractions E were separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in order and pooling fractions with identical thin layer behavior to give four sub-fractions E1-E4. E2 was further fractionated by silica gel column chromatography under reduced pressure using a gradient of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) in nine gradient systems in order and pooling fractions with identical thin layer behavior to give three sub-fractions E2-a-E2-c. E2-c was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in order and pooling fractions with identical thin layer behavior to give three fractions E2-c1-E2-c3. E2-c2 was purified by semi-preparative HPLC (acetonitrile-water, 55:45, 3 mL / min) to give compound 38 (8.8 mg). E3 was first separated by Sephadex LH-20 gel column chromatography (dichloromethane-methanol, 1:1) to give three sub-fractions E3-a-E3-c. E3-c was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in order and pooling fractions with identical thin layer behavior to give four fractions E3-c1-E3-c4. E3-c2 was purified by semi-preparative HPLC (acetonitrile-water, 60:40, 3 mL / min) to give compound 2* (4.6 mg), 3* (6.3 mg), 39 (7.8 mg). E3-c3 was purified by semi-preparative HPLC (acetonitrile-water, 65:35, 3 mL / min) to give compound 4* (7.5 mg). E4 was further fractionated by silica gel column chromatography under reduced pressure using a gradient of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) in nine gradient systems in order and pooling fractions with identical thin layer behavior to give three sub-fractions E4-a-E4-c. E4-b was separated by ODS reverse phase column chromatography eluting with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) in order and pooling fractions with identical thin layer behavior to give four fractions E4-b1-E4-b4. E4-b3 was purified by semi-preparative HPLC (acetonitrile-water, 70:30, 3 mL / min) to give compound 5* (7.5 mg) and 6* (8.5 mg).

[0059] Fractions F were re-segmented by flash column chromatography on silica gel with a gradient of petroleum ether-ethyl acetate (90:1, 70:1, 50:1, 30:1, 20:1, 10:1, 5:1, 1:1 and 0:1, v / v) to give three sub-fractions F1-F3 based on their TLC behavior. F3 was separated by ODS reverse phase column chromatography with a gradient of methanol / water (10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, v / v) to give three fractions F3-a-F3-c. F3-b was purified by semi-preparative HPLC (acetonitrile-water, 60:40, 3 mL / min) to give compound 7* (7.6 mg).

[0060] List of Urushiol Compounds Isolated from Dry Lac

[0061]

[0062] Structure Elucidation of Compound 1*

[0063] Compound 1* was obtained as a yellow oil. HRESIMS gave the pseudomolecular ion peak at m / z 275.2009 [M-H] – (calcd for C 18 H 27 O2, 275.2011), combined with 13 C NMR data, determined its molecular formula as C 18 H 28 O2, with five degrees of unsaturation. 1D NMR combined with HSQC (Table 1) analysis gave a signal of an ortho-trisubstituted benzene ring [δ H 6.70 (3H, overlapped, H-4,5,6) / δ C 122.2, 120.2, 113.0; δ C 129.5, 141.9, 143.2], a set of methylene long carbon chain signals [δ H 1.21-1.35 (12H, m, H-3',4',5',6',7',8') / δ C 29.3, 29.6, 29.6, 29.6, 29.7, 29.7; δ H 1.60 (2H, m, H-2') / δ C 29.7; δ H 1.96 (2H, m, H-9') / δ C 32.7; δ H 2.59 (2H, t, J = 7.8 Hz, H-1') / δ C29.9], two olefinic signals [δ H 5.41 (2H, m, H-10', 11') / δ C 124.6, 131.8] and one methyl signal [δ H 1.64 (3H, m, H-12') / δ C 18.1]. These signals are characteristic of catechol-type urushiol compounds. 1 H- 1 H COSY spectrum (Fig. 2), Figure 1 ), two olefinic signals (δ H 5.41) were correlated with H-9' and H-12', suggesting the double bond located at C-10' and C-11' of the long carbon chain. In the HMBC spectrum (Fig. 3), Figure 1 ), H-9' (δ H 1.96) was correlated with C-8', C-10' and C-11', and H-12' (δ H 1.64) was correlated with C-10' and C-11', further confirming the double bond located at C-10' and C-11'; meanwhile, the chemical shifts of C-9' (δ C 32.7) and C-12' (δ C 18.1) determined the double bond as trans configuration. In addition, EI-MS analysis revealed the key molecular fragments m / z 55.05 and 221.17 (Fig. 4), Figure 2 ), further confirming the double bond located at C-10' and C-11' of the long carbon chain. Finally, the HMBC correlation of H-1' (δ H 2.59) with C-2, C-3, C-4 and C-2' proved the long carbon chain attached at C-3 of the benzene ring. Therefore, compound 1* was identified as (E)-3-(dodec-10-enyl)benzene-1,2-diol, as shown in Fig. 5, named as toxicodenol A. Figure 1

[0064] Table 1 H (500 MHz) and C (125 MHz) NMR data of compound 1* (test solvent: CDC13; δ: ppm) 1 13

[0065]

[0066] a The assignments could be interchangeable.

[0067] Structure identification of compound 2* (Toxicodenol B) ​​​

[0068] Compound 2*was a yellow oil. HRESIMS gave the pseudomolecular ion peak m / z 263.1651 [M-H] – (calcd for C 16 H 23 O3,263.1647), combined 13 C NMR data, the molecular formula was determined as C 16 H 24 O3, with 5 degrees of unsaturation. 1D NMR combined with HSQC (Table 2) analysis gave one ortho-trisubstituted benzene ring signal [δ H 6.70 (3H, overlapped, H-4,5,6) / δ C 122.1, 120.1, 113.0; δ C 129.5, 142.2, 143.3], one set of methylene long carbon chain signals [δ H 1.24-1.35 (8H, m, H-3',4',5',6') / δ C 29.5, 29.5, 29.2, 29.3; δ H 1.55 (2H, m, H-7') / δ C 23.9; δ H 1.61 (2H, m, H-2') / δ C 29.8; δ H 2.41 (2H, t, J = 7.4 Hz, H-8') / δ C 43.9; δ H 2.60 (2H, t, J = 8.0 Hz, H-1') / δ C 29.9], one methyl proton signal [δ H 2.14 (3H, s, H-10') / δ C 30.1] and one carbonyl carbon signal (δ C 210.5), which are characteristic signals of ortho-diphenol type urushiol compounds. In the HMBC spectrum ( Figure 3 ), H-8' (δ H 2.41, t, J = 7.4 Hz) was correlated to C-7', C-6' and C-9' (δ C 210.5), H-10' (δ H 2.14, 3H, s) was also correlated to C-9', which proved the substitution of carbonyl group at C-9'. Finally, H-1' (δ H2.60, t, J = 8.0 Hz) and HMBC signals associated with C-2, C-3, C-4 and C-2' proved the long carbon chain attached at C-3 of the phenyl ring. Thus, compound 2* was identified as 10-(2,3-dihydroxyphenyl)decan-2-one, as shown in Figure 3 Figure 1, named toxicodenol B.

[0069] Table 2.1D NMR data of compounds 2* and 3* 1 H (500 MHz) and 13 C (125 MHz) NMR data (test solvent: CDC13; δ: ppm)

[0070]

[0071] a The assignments could be interchangeable

[0072] Structure identification of compound 3* (Toxicodenol C)

[0073] Compound 3* was a yellow oil. HRESIMS gave a quasi-molecular ion peak at m / z 277.1808 [M-H] – (calcd for C 17 H 25 O3, 277.1804), combined with 13 C NMR data, determined its molecular formula as C 17 H 26 O3, with 5 degrees of unsaturation. The 1D NMR of compound 3* was similar to that of compound 2*, with the main difference being the number of methylene long carbon chains. In compound 3*, 1D NMR combined with HSQC (Table 2) analysis also gave a signal of an ortho-trisubstituted phenyl ring [δ H 6.69 (3H, overlapped, H-4,5,6) / δ C 121.0, 120.0, 113.0; δ C 129.5; δ C 142.3; δ C 143.4], a set of methylene long carbon chain signals [δ H 1.24-1.34 (10H, m, H-3',4',5',6',7') / δ C 29.5, 29.5, 29.4, 29.2, 29.5; δ H 1.55 (2H, m, H-8') / δ C 23.9; δ H1.59 (2H, m, H-2') / δ C 29.8; δ H 2.42 (2H, t, J = 7.4 Hz, H-9') / δ C 43.9; δ H 2.59 (2H, t, J = 7.8 Hz, H-1') / δ C 29.9], one methyl proton signal [δ H 2.14 (3H, s, H-10') / δ C 30.0] and one carbonyl carbon signal (δ C 210.5), which are characteristic signals of catechol-type urushiol compounds. In the HMBC spectrum ( Figure 4 ), H-9' (δ H 2.42, t, J = 7.4 Hz) is correlated to C-8', C-7' and C-10' (δ C 210.5), H-11' (δ H 2.14, 3H, s) is correlated to C-10', which confirms that the carbonyl group is located at C-10' of the long carbon chain. Finally, the HMBC signal of H-1' (δ H 2.59, t, J = 7.8 Hz) is correlated to C-2, C-3, C-4 and C-2', which confirms that the long carbon chain is attached at C-3 of the benzene ring. Compound 3* is identified as 11-(2,3-dihydroxyphenyl)undecan-2-one, as shown in Figure 4 , named as toxicodenol C.

[0074] Structural identification of compound 5* (Toxicodenol E)

[0075] Compound 5* is a yellow oil. HRESIMS gives the quasi-molecular ion peak m / z 361.2749 [M-H] – (calcd for C 23 H 37 O3, 361.2743), combined with 13 C NMR data, confirms its molecular formula as C 23 H 38 O3, with 5 degrees of unsaturation. 1D NMR combined with HSQC (Table 3) analysis gives one ortho-trisubstituted benzene ring signal [δ H 6.68 (3H, overlapped, H-4, 5, 6) / δ C 121.9, 119.9, 112.9; δ C 129.5; δ C 142.3; δ C143.4], a set of methylene long carbon chain signals [δ H 1.26-1.35(18H,m,H-3′,4′,5′,6′,7′,8′,14′,15′,16′) / δ C 29.7,29.6,29.5,29.4,29.0,25.4,29.3,31.8,22.8; δ H 1.59(2H,m,H-2′) / δ C 29.8; δ H 1.59(2H,m,H-9′) / δ C 37.5; δ H 2.05(2H,m,H-13′) / δ C 27.8; δ H 2.59(2H,t,J=7.5Hz,H-1′) / δ C 29.9], two olefin signals [δ H 5.35(1H,m,H-11′) / δ C 132.2; δ H 5.49(1H,m,H-12′) / δ C 132.9], an oxygen-containing methine proton signal [δ H 4.45(1H,m,H-10′) / δ C 68.2] and a methyl proton signal [δ H 0.88(3H,m,H-17′) / δ C [14.2] is a characteristic signal of catechol-type urushiol compounds. 1 H- 1 In the H COSY spectrum ( Figure 5 ), the proton signals of two olefinic properties [δ H 5.35 (1H, m, H-11′); δ H 5.49(1H,m,H-12′)] and H-10′(δ H 4.45, 1H, m) and H-13′ (δ H The correlation between 2.05, 2H, and m suggests that the double bond is connected to an oxygen-containing methine group. In the HMBC spectrum ( Figure 5 The correlation between H-10′ and C-8′, C-9′, and C-12′, and between H-13′ and C-11′ and C-14′, further confirms that the double bond is linked to an oxygen-containing methine group. Additionally, based on C-13′ (δ... C The chemical shift at 27.8) confirmed the double bond to be in cis configuration. In the NOE spectrum, H-10′(δ) H 4.45, 1H, m) and H-13′ (δ HThe correlation between 2.05, 2H, and m also confirmed the cis configuration of the double bond. To determine the position of the double bond in the long carbon chain, we used HREI-MS analysis and found that the key molecular fragments had m / z values ​​of 97.10119, 123.04397, and 291.42780 (m / z). Figure 6 This indicates that the double bond is located at C-11′ and C-12′ of the long carbon chain. Combined with high-resolution numerical analysis, it is deduced that the oxygen-containing substituent at C-10′ is a hydroxyl group. Finally, through H-1′ (δ... H 2.59) The correlation between the HMBCs of C-2, C-3, C-4, and C-2' confirms that a long carbon chain is attached to the C-3 position of the benzene ring. The optical rotation of compound 5* was measured to be 0, indicating that it is a racemic mixture. Unfortunately, it could not be resolved by chiral column chromatography, making separation difficult. Therefore, compound 5* was identified as (Z)-3-(10-hydroxyheptadec-11-enyl)benzene-1,2-diol, as... Figure 5 As shown, it is named toxicodenol D.

[0076] Table 3 Compounds 5* and 6* 1 H (500MHz) and 13 C (125MHz) NMR data (test solvent: CDCl3; δ: ppm)

[0077]

[0078] Structural identification data of compound 6* (Toxicodenol F)

[0079] Compound 6* is a yellow oily substance. HRESIMS gives it a quasi-molecular ion peak at m / z 379.2853 [M–H]. – (calcdfor C 23 H 39 O4,379.2847), combined with 13 C10 NMR data determined its molecular formula to be C10. 23 H 40 O4, with an unsaturation degree of 4. 1D NMR combined with HSQC (Table 3) analysis showed a signal of an ortho-trisubstituted benzene ring [δ]. H 6.69(3H,overlapped,H-4,5,6) / δ C 122.1, 120.1, 113.0; δ C 129.5; δ C 142.2; δ C 143.3], a set of methylene long carbon chain signals [δ H1.25-1.34 (20H, m, H-3', 4', 5', 6', 7', 8', 13', 14', 15', 16') / δ C 29.8, 29.8, 29.8, 29.5, 29.3, 25.6, 25.6, 29.3, 31.9, 22.8; δ H 1.60 (2H, m, H-2') / δ C 29.6; δ H 1.46 (4H, m, H-9', H-12') / δ C 33.6; δ H 2.60 (2H, t, J = 7.3 Hz, H-1') / δ C 29.8], two oxymethylene proton signals [δ H 3.43 (2H, m, H-10', 11') / δ C 74.7] and one methyl proton signal [δ H 0.88 (3H, t, J = 6.2 Hz, H-17') / δ C 14.3], are characteristic signals of catechol-type urushiborols. 1 H- 1 H COSY spectrum ( Figure 7 ), two oxymethylene proton signals [δ H 3.43 (2H, m, H-10', 11')] are correlated with H-9' and H-12' (δ H 1.46, 4H, m), combined with the high-resolution data, we deduced the presence of vicinal diol structure on the long carbon chain. To determine the position of the vicinal diol on the long carbon chain, we used EI-MS analysis, and found the key molecular fragments m / z 115.16, 123.20 and 265.44 ( Figure 8 ), which indicated that the vicinal diol was located at C-10' and C-11' of the long carbon chain. Finally, the HMBC correlation of H-1' (δ H 2.60) with C-2, C-3, C-4 and C-2' proved that the long carbon chain was attached at C-3 of the benzene ring. The optical rotation value of compound 6* was 0, indicating that it was a racemate, unfortunately, we failed to separate it by chiral column, which was difficult to separate. Therefore, compound 6* was identified as 3-(10, 11-dihydroxyheptadecyl) benzene-1, 2-diol, as shown in Figure 7 , named as toxicodenol F.

[0080] Structure identification of compound 8* (Toxicodenol H)

[0081] Compound 8* was a yellow oil. HRESIMS gave the pseudo-molecular ion peak m / z 311.2017 [M-H] – (calcd for C 21 H 27 O2,311.2011), combined with 13 C NMR data, determined its molecular formula as C 21 H 28 O2, with 8 unsaturations. 8* was similar to 7* in 1D NMR, the main difference between the two compounds was that 8* had one more methylene in the long carbon chain. In compound 8*, 1D NMR combined with HSQC analysis gave one ortho-trisubstituted benzene ring signal [δ H 6.71 (3H, overlapped, H-4,5,6) / δ C 122.1, 120.1, 113.0; δ C 129.5; δ C 142.1; δ C 143.2] and a set of 9 methylene long carbon chain signals [δ H 1.29-1.34 (10H, m, H-3",4",5",6",7") / δ C 29.6, 29.7, 29.7, 29.6, 29.5; δ H 1.60 (4H, m, H-2", H-8") / δ C 31.7; δ H 2.60 (4H, t, J = 7.5 Hz, H-1",9") / δ C 29.9, 36.1]. Unlike the characteristic signals of ortho-diphenol type urushiol compounds, it did not have long carbon chain terminal methyl proton signals, but had one more monosubstituted benzene ring signal [δ H 7.17 (1H, m, H-4') / δ C 125.7; δ H 7.19 (2H, m, H-3',5') / δ C 128.5; δ H 7.29 (2H, m, H-2',6') / δ C 128.3; δ C 143.1]. In the HMBC spectrum ( Figure 9 ), H-1" (δ H 2.60, t, J = 7.5 Hz) was correlated to C-3, C-4 and C-2", H-9" (δ H2.60, t, J = 7.5 Hz) associated with C-1', C-2' and C-6' proved the long carbon chain with two benzene rings at both ends, one at C-3 and the other at C-1' of the benzene ring, respectively. Thus, compound 8* was identified as 3-(9-phenylnonyl)benzene-1,2-diol, as shown in Figure 9 Figure 1, named toxicodenol H.

[0082] Structure identification of compound 9* (Toxicodenol I)

[0083] Compound 9* was a yellow oil. HRESIMS gave the quasi-molecular ion peak m / z 339.2325 [M - H] – (calcd for C 23 H 31 O2, 339.2324), combined with 13 C NMR data, determined its molecular formula as C 23 H 32 O2, with 8 degrees of unsaturation. 1D NMR combined with HSQC (Table 4) analysis gave a signal of an ortho-trisubstituted benzene ring [δ H 6.71 (3H, overlapped, H-4,5,6) / δ C 122.2, 120.2, 113.0; δ C 129.5; δ C 142.1; δ C 143.2] and a set of 10 methylene long carbon chain signals [δ H 1.26-1.37 (12H, m, H-3",4",5",6",7",8") / δ C 29.8, 29.8, 29.8, 29.7, 29.7, 29.7; δ H 1.56 (2H, m, H-9") / δ C 30.4; δ H 1.60 (2H, m, H-2") / δ C 29.9; δ H 2.58 (2H, m, H-10") / δ C 33.5; δ H 2.60 (2H, m, H-1") / δ C 29.8] and a methyl proton signal [δ H 2.30 (3H, s, H3-2') / δ C 19.4]. Different from the characteristic signals of the above catechol-type urushols, it had one more disubstituted benzene ring signal [δ H7.12(4H,m,H-3′,4′,5′,6′) / δ C 130.2, 125.8, 125.9, 128.9; δ C 136.0; δ C 141.3]. In the HMBC spectrum ( Figure 10 ), H-1″(δ H 2.60,m) is associated with C-2, C-4 and C-2″, H-10″ (δ H 2.58,m) is associated with C-1′, C-2′, and C-6′, proving that the long carbon chain is connected to a benzene ring at each end, located at C-3 and C-1′ of the benzene ring, respectively. Meanwhile, H3-2′ (δ H The HMBC correlation of 2.30,s) with C-1′, C-2′, C-3′, and C-10″ determined that the methyl group was located at C-2′ of the benzene ring. Therefore, compound 9* was identified as 3-(10-(o-tolyl)decyl)benzene-1,2-diol, as... Figure 10 As shown, it is named toxicodenol I.

[0084] Table 4. Compounds 9* and 10* 1 H (500MHz) and 13 C (125MHz) NMR data (test solvent: CDCl3; δ: ppm)

[0085]

[0086] Continued from Table 4 for compounds 9* and 10* 1 H (500MHz) and 13 C (125MHz) NMR data (test solvent: CDCl3; δ: ppm)

[0087]

[0088] Structural identification of compound 10* (Toxicodenol J)

[0089] Compound 10* is a yellow oily substance. HRESIMS gives it a quasi-molecular ion peak at m / z 339.2328 [M–H]. – (calcdfor C 23 H 31 O2,339.2324), combined 13 C10 NMR data determined its molecular formula to be C10. 23 H 32 O2, with an unsaturation degree of 8. 1D NMR combined with HSQC (Table 4) analysis yielded a signal of an ortho-trisubstituted benzene ring [δ]. H6.71(3H,overlapped,H-4,5,6) / δ C 122.2, 120.2, 113.0; δ C 129.2; δ C 140.6; δ C 142.0] and a set of 9 methylene long carbon chain signals [δ H 1.27-1.37(10H,m,H-3″,4″,5″,6″,7″) / δ C 29.7,29.7,29.7,29.7,29.9; δ H 1.58(4H,m,H-2″,8″) / δ C 29.9; δ H 2.60(4H,m,H-1″,9″) / δ C 29.9]. Unlike the characteristic signals of catechol-type urushiol compounds, it has an additional signal of a disubstituted benzene ring [δ]. H 7.13(4H,m,H-3′,4′,5′,6′) / δ C 128.4, 126.0, 125.9, 129.5; δ C 141.4; δ C 141.9] and an ethyl signal [δ H 2.66(2H,q,J=7.6Hz) / δ C 25.6; δ H 1.22(3H,t,J=7.6Hz) / δ C 15.5]. 1 H- 1 In the H COSY spectrum ( Figure 11 ), the methylene proton signal of the ethyl group [δ H 2.66(2H,q,J=7.6Hz) / δ C 25.6] and methyl proton signal [δ H 1.22(3H,t,J=7.6Hz) / δ C [15.5] Related, verifying the presence of ethyl substitution in the compound. In the HMBC spectrum ( Figure 11 ), methylene proton signal [δ H 2.66(2H,q,J=7.6Hz) / δ C [25.6] Related to C-2′ and C-3′, methyl proton signal [δ] H 1.22(3H,t,J=7.6Hz) / δ C [15.5] is also related to C-2′, confirming that the ethyl group is located at C-2′ of the benzene ring. Finally, H-1″(δ H2.60, m) associated with C-3, C-4 and C-2", H-9" (δ H 2.60, m) associated with C-1' and C-8", indicating a long carbon chain with a benzene ring at each end, and located at C-3 and C-1', respectively. Thus, compound 10* was identified as 3-(9-(2-ethylphenyl)nonyl)benzene-1,2-diol, as shown, named toxicodenol J. Figure 11

[0090] Structure identification of compound 12* (Toxicodenol L)

[0091] Compound 12* was a yellow oil. HRESIMS gave the quasi-molecular ion peak m / z 353.2124 [M-H] – (calcd for C 23 H 29 O3, 353.2117), combined with 13 C NMR data, determined its molecular formula as C 23 H 30 O3, with 9 degrees of unsaturation. 1D NMR combined with HSQC analysis gave a signal of an ortho-trisubstituted benzene ring [δ H 6.71 (3H, overlapped, H-4,5,6) / δ C 122.2, 120.2, 113.0; δ C 129.2; δ C 142.1; δ C 143.2], a set of methylene long carbon chain signals [δ H 1.25-1.35 (10H, m, H-3",4",5",6",7") / δ C 29.4, 29.6, 29.6, 29.5, 29.4; δ H 1.60 (2H, m, H-2") / δ C 29.9; δ H 1.68 (2H, m, H-8") / δ C 24.5; δ H 2.60 (2H, t, J = 7.5 Hz, H-1") / δ C 29.9; δ H 2.88 (2H, t, J = 7.4 Hz, H-9") / δ C 41.8]. Unlike the characteristic signals of the ortho-diphenol type urushiol compounds, it has no long carbon chain end methyl proton signal, but has an additional disubstituted benzene ring signal [δ H ​7.16 (1H, d, J = 7.8 Hz, H-6') / δ C 128.4; δ H 7.25 (1H, d, J = 7.8 Hz, H-3') / δ C 132.0; δ H 7.30 (1H, t, J = 7.8 Hz, H-5') / δ C 125.8; δ H 7.36 (1H, t, J = 7.8 Hz, H-4') / δ C 131.2; δ C 137.9; δ C 138.4], one methyl proton signal [δ H 2.48 (3H, s) / δ C 21.4] and one carbonyl signal (δ C 205.5). In the HMBC spectrum ( Figure 12 ), H-1" (δ H 2.60) is correlated to C-2, C-3, C-4 and C-2", which proves that the long carbon chain is connected to the C-3 position of the ortho-diphenol benzene ring at one end. Meanwhile, H-8", H-9" and H-6' are all HMBC correlated to the carbonyl (δ C 201.1), which deduces that the carbonyl is located at the C-10" position of the long carbon chain and is connected to the C-1' position of the disubstituted benzene ring. Finally, the HMBC correlation of the methyl proton signal [δ H 2.48 (3H, s)] to C-1', C-3' indicates that the methyl is located at the C-2' position of the benzene ring. Therefore, compound 12* is identified as 10-(2,3-dihydroxyphenyl)-1-(o-tolyl)decan-1-one, as shown in Figure 12 , named as toxicodenol L.

[0092] Structure identification of compound 14* (Toxicodenol N)

[0093] Compound 14* is a yellow oil. HRESIMS gives the quasi-molecular ion peak m / z 263.1651 [M - H] – (calcd for C 16 H 23 O3, 263.1647), combined with 13 C NMR data, determines its molecular formula as C 16 H 24 O3, with 5 degrees of unsaturation. 1D NMR combined with HSQC (Table 5) analysis gives one meta-substituted benzene ring signal [δ H6.65(1H,d,J=7.7Hz,H-6) / δ C 112.7; δ H 6.67(1H,s,H-2) / δ C 115.4; δ H 6.74(1H,d,J=7.7Hz,H⁻⁴) / δ C 121.0; δ H 7.13(1H,t,J=7.7Hz,H⁻⁵) / δ C 129.5; δ C 144.8; δ C 155.7], a set of methylene long carbon chain signals [δ H 1.30-1.32(6H,m,H-3',H-4',H-5') / δ C 29.1,29.1,29.1; δ H 1.60(4H,m,H-2',H-6') / δ C 31.2, 25.0; δ H 2.29(2H,t,J=7.5Hz,H-7') / δ C 34.5; δ H 2.55(2H,t,J=7.7Hz,H⁻¹') / δ C 35.8], a carbonyl signal (δ C 174.3, C-8'), a hydroxymethylene proton signal [δ H 4.14(2H,q,J=7.1Hz,H-9') / δ C 60.4] and a methyl proton signal [δ H 1.25(3H,t,J=7.1Hz) / δ C 14.4,H-10'] is a characteristic signal of monophenolic compounds. 1 H- 1 In the H COSY spectrum ( Figure 13 ), δ-oxymethylene proton signal H 4.14) and methyl proton signal (δ H Based on the correlation with 1.25 and combined with high-resolution numerical values, the C-9′ terminal of the long carbon chain was determined to be an oxyethyl group. In the HMBC spectrum ( Figure 13 ), H-9′(δ H 4.14) and H-7′(δ H 2.29) are both related to carbonyl (δ) C (174.3) Related evidence shows that the carbonyl group is located at the C-8′ position of the long carbon chain, and the ethoxy group is connected to the long carbon chain through the carbonyl group. Finally, through H-1′ (δ H2.55) HMBC correlation with C-2, C-3, C-4 and C-2' proved the long carbon chain was attached at the C-3 position of the benzene ring. Thus, compound 14* was identified as ethyl 8-(3-hydroxyphenyl)octanoate, as shown in Figure 13 Figure 2, named toxicodenol N. LC / MS Figure 14 ) was further used to confirm 14* as a natural product, not an artificial product.

[0094] Table 5 H (500 MHz) and 1 C (125 MHz) NMR data of compound 14* (test solvent: CDC13; δ: ppm) 13

[0095]

[0096] 2.3.16 Structure identification of compound 16* (Toxicodenol P)

[0097] Compound 16* was a yellow oil. HRESIMS gave the quasi-molecular ion peak m / z 317.2480 [M-H] – (calcd for C 21 H 33 O2, 317.2481), combined with 13 C NMR data, determined its molecular formula as C 21 H 34 O2, with 5 degrees of unsaturation. 1D NMR combined with HSQC (Table 6) analysis gave one meta-substituted benzene ring signal [δ H 6.64 (1H, d, J = 7.7 Hz, H-6) / δ C 112.7; δ H 6.66 (1H, s, H-2) / δ C 115.4; δ H 6.72 (1H, d, J = 7.7 Hz, H-4) / δ C 120.7; δ H 7.13 (1H, t, J = 7.7 Hz, H-5) / δ C 129.4; δ C 144.8; δ C 155.9], and one set of methylene long carbon chain signals [δ H 1.30-1.35 (12H, m, H-3', H-4', H-5', H-6', H-13', H-14') / δ C 22.8, 27.9, 29.1, 29.2, 29.3, 29.6; δ H ​1.49(2H,m,H-12′) / δ C 36.4; δ H 1.59(2H,m,H-2′) / δ C 31.2; δ H 2.04(2H,q,J=6.8Hz,H-7′) / δ C 27.5; δ H 2.23(2H,t,J=6.6Hz,H-10′) / δ C 35.5; δ H 2.25(2H,t,J=7.5Hz,H-1′) / δ C 35.8], two olefin signals [δ H 5.41(1H,m,H-9′) / δ C 125.1; δ H 5.56(1H,m,H-8′) / δ C 133.8], a hydroxymethyl proton signal [δ H 3.65(1H,m,H-11′) / δ C 71.9] and a methyl proton signal [δ H 0.90(3H,t,J=6.9Hz,H-15′) / δ C [14.2] is a characteristic signal of monophenolic compounds. 1 H- 1 In the H COSY spectrum ( Figure 14 ), the proton signal (δ) of two olefinic properties H 5.41, 5.56) are related to H-7′ and H-10′, according to C-7′(δ C The chemical shift at 27.5) confirmed the double bond to be in the cis configuration. Simultaneously, the hydroxymethyl (δ) H 3.65) and H-10'(δ) H 2.23) and H-12'(δ H 1.49) all have COSY correlation. In the HMBC spectrum ( Figure 15 ), H-10'(δ H 2.23) and C-8'(δ) C 133.8), C-9'(δ) C 125.1), C-11'(δ C 71.9) and C-12'(δ C36.4) are all relevant, proving that the oxymethimide and the double bond are located adjacent to each other on the long carbon chain. HREI-MS analysis yielded key molecular fragments 87.8051, 108.05687, and 261.18503, determining the double bond position to be at C-8', 9'. Combined with high-resolution numerical values ​​and chemical shift values ​​[δ...], H 3.65(1H,m,H-11′) / δ C 71.9], determine if there is a hydroxyl substitution at C-11' ( Figure 15 Finally, through H-1′(δ) H 2.55) correlated with the HMBCs of C-2, C-3, C-4, and C-2', confirming the long carbon chain attached to the C-3 position of the benzene ring. The optical rotation of compound 16* was measured to be 0, indicating it is a racemic mixture. Unfortunately, it could not be resolved by chiral column chromatography, making separation difficult. Therefore, compound 16* was identified as (Z)-3-(11-hydroxypentadec-8-enyl)phenol, as... Figure 14 As shown, it is named toxicodenol Q.

[0098] Table 6 Compound 16* 1 H (500MHz) and 13 C (125MHz) NMR data (test solvent: CDCl3; δ: ppm)

[0099]

[0100] Structural identification of compound 18* (Toxicodenol R)

[0101] Compound 18* is a yellow oily substance. HRESIMS gives it a quasi-molecular ion peak at m / z 329.2484 [M–H]. – (calcdfor C 22 H 33 O2,329.2481), combined 13 C10 NMR data determined its molecular formula to be C10. 22 H 34 O2, with an unsaturation degree of 6. 1D NMR combined with HSQC analysis yielded a signal for a benzene ring with 1, 2, 4-three-position substitution [δ]. H 6.80(1H,s,H-3) / δ C 117.2; δ H 6.83(1H,d,J=8.0Hz,H⁻⁵) / δ C 120.7; δ H 7.45 (1H,d,J=8.0Hz,H⁻⁶) / δ C 133.8; δ C118.9; δ C 153.9; δ C 161.9], a set of methylene long carbon chain signals [δ H 1.25-1.36(16H,m,H-3′,H-4′,H-5′,H-10′,H-11′,H-12′,H-13′,H-14′) / δ C 22.8,29.1,29.3,29.3,29.5,29.8,29.8,31.9; δ H 1.61(2H,m,H-2′) / δ C 30.8; δ H 2.01(4H,m,H-6′,H-9′) / δ C 27.3, 27.4; δ H 2.61(2H,t,J=7.8Hz,H-2′) / δ C 36.6], two olefin proton signals [δ H 5.34(2H,m,H-7′,H-8′) / δ C 129.9, 130.1], an aldehyde matrix sub-signal [δ H 9.83(1H,s) / δ C 196.0], a methyl proton signal [δ H 0.88(3H,t,J=6.5Hz,H-15′) / δ C 14.3] and an active hydrogen signal (δ H 11.06,s) is a characteristic signal of urushiol compounds. 1 H- 1 In the H COSY spectrum ( Figure 16 ), the proton signal (δ) of two olefinic properties H 5.34) is related to H-6′ and H-9′, according to C-6′(δ C 27.3) and C-9'(δ C The chemical shift at 27.4) confirmed the double bond to be in cis configuration. In the HMBC spectrum ( Figure 16 ), aldehyde matrix signal (δ H 9.83) is correlated with C-1, and the active hydrogen signal (δ) H 11.06) is associated with C-1, C-2 and C-3, H-1′(δ) H2.61) and C-3, C-4, C-5 and C-2', it was deduced that the aldehyde group, the hydroxyl group and the long carbon chain were located at C-1, C-2 and C-4 of the benzene ring, respectively. Finally, the double bond was determined to be located at C-7' and C-8' by HREI-MS analysis of key molecular fragments m / z 136.05197, 191.10670, 231.13831. Figure 17 ). Therefore, compound 18* was identified as (Z)-2-hydroxy-4-(pentadec-7-enyl)benzaldehyde, as shown in Figure 16 , named as toxicodenol R.

[0102] Structure identification of compound 19* (Toxicodenol S)

[0103] Compound 19* was a yellow oil. HRESIMS gave the quasi-molecular ion peak m / z 373.2744 [M-H] – (calcd for C 24 H 37 O3, 373.2743), combined with 13 C NMR data, determined its molecular formula as C 24 H 38 O3, with 6 degrees of unsaturation. 1D NMR combined with HSQC (Table 7) analysis gave a signal of a 1,2,3,4-tetra-substituted benzene ring [δ H 6.81 (1H, d, J = 8.0 Hz, H-6) / δ C 121.4; δ H 7.06 (1H, d, J = 8.0 Hz, H-5) / δ C 123.9; δ C 118.8, 137.6, 142.4, 148.0], a set of methylene long carbon chain signals [δ H 1.25-1.36 (20H, m, H-3', H-4', H-5', H-10', H-11', H-12', H-13', H-14', H-15', H-16') / δ C 22.8, 29.2, 29.7, 29.7, 29.7, 29.7, 29.7, 29.8, 29.9, 31.9; δ H 1.61 (2H, m) / δ C 29.4; δ H 2.01 (4H, q, J = 5.7 Hz) / δ C 27.3, 27.4; δ H 2.61 (2H, t, J = 7.8 Hz, H-1') / δ C30.5], two olefin proton signals [δ H 5.34(2H,m,H-7′,H-8′) / δ C 130.0, 130.1], an aldehyde matrix sub-signal [δ H 9.82(1H,s) / δ C 196.4], a methyl proton signal [δ H 0.88(3H,t,J=6.8Hz) / δ C 14.3] and two active hydrogen signals (δ H 5.63 s; δ H 11.13,s) is a characteristic signal of urushiol compounds. 1 H- 1 In the H COSY spectrum ( Figure 19 ), the proton signal (δ) of two olefinic properties H 5.34) is related to H-6′ and H-9′, according to C-6′(δ C 27.3) and C-9'(δ C The chemical shift at 27.4) confirmed the double bond to be in cis configuration. In the HMBC spectrum ( Figure 18 ), aldehyde matrix signal (δ H 9.82) is correlated with C-1 and C-2, and the active hydrogen signal (δ) H 11.13) is associated with C-1, C-2, and C-3, and another active hydrogen signal (δ) H 5.63) is associated with C-2, C-3 and C-4, H-1′(δ) H 2.01) was correlated with C-3, C-4, C-5, and C-2', leading to the deduction that the aldehyde group, two hydroxyl groups, and long carbon chain are located at C-1, C-2, C-3, and C-4 positions on the benzene ring, respectively. Finally, EI-MS analysis of key molecular fragments (m / z 153.13, 221.22, and 247.14) confirmed that the double bonds are located at C-7' and C-8'. Figure 19 Therefore, compound 19* was identified as (Z)-4-(heptadec-7-enyl)-2,3-dihydroxybenzaldehyde, as... Figure 18 As shown, it is named toxicodenol S.

[0104] Table 7 Compound 19* 1 H (500MHz) and 13 C (125MHz) NMR data (test solvent: CDCl3; δ: ppm)

[0105]

[0106] Structure identification of compound 20*

[0107] Compound 20*was a yellow oil. HRESIMS gave a pseudomolecular ion peak at m / z 387.2901 [M-H] – (calcd for C 25 H 39 O3,387.2899), combined with 13 C NMR data, determined its molecular formula as C 25 H 40 O3, with six degrees of unsaturation. 1D NMR combined with HSQC (Table 8) analysis gave a 1,2,3,4-tetra-substituted benzene ring signal [δ H 6.69 (1H, d, J = 8.3 Hz, H-6) / δ C 120.1; δ H 7.21 (1H, d, J = 8.3 Hz, H-5) / δ C 120.9; δ C 117.7; δ C 136.2; δ C 143.0; δ C 149.2], a set of methylene long carbon chain signals [δ H 1.25-1.35 (20H, m, H-3', H-4', H-5', H-6', H-7', H-8', H-9', H-10', H-15', H-16') / δ C 22.8, 29.1, 29.4, 29.6, 29.7, 29.8, 29.8, 29.8, 29.9, 31.9; δ H 1.61 (2H, m, H-2') / δ C 29.6; δ H 2.01 (4H, q, J = 5.7 Hz, H-11', H-14') / δ C 27.3, 27.3; δ H 2.66 (2H, t, J = 7.8 Hz, H-1') / δ C 30.3], two olefinic proton signals [δ H 5.35 (2H, m, H-12', H-13') / δ C 130.0, 130.1], one carbonyl signal (δ C 204.6), two methyl proton signals [δ H 0.88 (3H, t, J = 6.1 Hz, H-17') / δ C 14.3; δ H 2.60 (3H, s) / δC 26.6] and two active hydrogen signals (δ H 5.75, s; δ H 12.52, s) are characteristic signals of urushiol compounds. 1 H- 1 HCOSY spectrum (Fig. 2) of 20*, Figure 20 ), two olefinic proton signals (δ H 5.35) are associated with H-11' and H-14', and the chemical shifts of C-11'(δ C 27.3) and C-14'(δ C 27.3) confirm the cis configuration of the double bond. In the HMBC spectrum (Fig. 3) of 20*, Figure 20 ), H-5 (δ H 7.21), H-6 (δ H 6.69) and a methyl proton signal (δ H 2.60) are all associated with the carbonyl group (δ C 204.6), indicating that the carbomethyl group is located at C-1 of the benzene ring. The active hydrogen signal (δ H 12.52) is associated with C-1, C-2 and C-3, and another active hydrogen signal (δ H 5.75) is associated with C-2, C-3 and C-4, H-1'(δ H 2.66) is associated with C-3, C-4 and C-2', it is deduced that the two hydroxyl groups and the long carbon chain are located at C-2, C-3 and C-4 of the benzene ring, respectively. Finally, the key molecular fragments m / z 43.01777, 166.06233, 331.22647 and 345.24213 determined by HREI-MS analysis confirm that the double bond is located at C-12' and C-13' ( Figure 21 ). Therefore, compound 20* is identified as (Z)-1-(4-(heptadec-12-enyl)-2,3-dihydroxyphenyl)ethan-1-one, as shown in Figure 20 , named as toxicodenol T.

[0108] Table 8 H (500 MHz) and 1 C (125 MHz) NMR data of compound 20* (test solvent: CDC13; δ: ppm) 13

[0109]

[0110] a The assignments could be interchangeable

[0111] ​Structure identification of compound 21*

[0112] Compound 21* was a yellow oil. HRESIMS gave the pseudomolecular ion peak at m / z 387.2900 [M-H] – (calcd for C 25 H 39 O3,387.2899), combined with 13 C NMR data, determined its molecular formula as C 25 H 40 O3, with 6 degrees of unsaturation. In compound 21*, 1D NMR combined with HSQC (Table 9) analysis gave a 1,2,3,4-tetra-substituted benzene ring signal [δ H 6.68 (1H, d, J = 8.2 Hz, H-6) / δ C 120.1; δ H 7.21 (1H, d, J = 8.2 Hz, H-5) / δ C 120.8; δ C 117.7; δ C 136.1; δ C 142.8; δ C 149.2], a set of methylene long carbon chain signals [δ H 1.25-1.35 (20H, m, H-3', H-4', H-5', H-6', H-7', H-8', H-9', H-10', H-15', H-16') / δ C 22.7, 29.1, 29.3, 29.6, 29.6, 29.6, 29.6, 29.6, 29.7, 31.8; δ H 1.61 (2H, m, H-2') / δ C 29.4; δ H 1.95 (4H, m, H-11', H-14') / δ C 32.7, 32.7; δ H 2.66 (2H, t, J = 7.7 Hz, H-1') / δ C 30.2], two olefinic proton signals [δ H 5.38 (2H, m, H-12', H-13') / δ C 130.4, 130.4], one carbonyl signal (δ C 204.5), two methyl proton signals [δ H 0.87 (3H, t, J = 6.1 Hz, H-17') / δ C 14.2; δ H 2.60 (3H, s) / δC 26.5] and two active hydrogen signals (δ H 5.75 s; δ H 12.52, s), is a characteristic signal of urushiol compounds. The 1D NMR spectra of 21* and 20* are similar; the main difference between the two compounds lies in the relative configuration of their double bonds. 1 H- 1 In the H COSY spectrum ( Figure 21 ), two olefin proton signals (δ H 5.38) is related to H-11′ and H-14′, according to C-11′(δ C 32.7) and C-14'(δ C The chemical shift at 32.7) confirmed the double bond to be in the trans configuration. In the HMBC spectrum ( Figure 22 H-6(δ) H 6.68) and methyl (δ) H 2.60) are both related to carbonyl (δ) C (204.5) is related, proving that the carbonyl methyl group is located at the C-1 position of the benzene ring. The active hydrogen signal (δ) H 12.52) is associated with C-1, C-2, and C-3, and another active hydrogen signal (δ) H 5.75) is associated with C-2, C-3 and C-4, H-1′(δ H 2.66) was associated with C-3, C-4, and C-2', leading to the deduction that the two hydroxyl groups and the long carbon chain were located at C-2, C-3, and C-4 positions on the benzene ring, respectively. Finally, EI-MS analysis of the key molecular fragments (m / z 57.07 and 83.10) confirmed that the double bonds were located at C-12' ​​and C-13'. Figure 23 Therefore, compound 21* was identified as (E)-1-(4-(heptadec-12-enyl)-2,3-dihydroxyphenyl)ethan-1-one, as... Figure 22 As shown, it is named toxicodenol U.

[0113] Table 9 Compound 21* 1 H (500MHz) and 13 C (125MHz) NMR data (test solvent: CDCl3; δ: ppm)

[0114]

[0115] a The assignments could be interchangeable

[0116] Compound 4. Toxicodenol D

[0117]

[0118] Yellow oil; UV (MeOH) λmax208, 240, 272 nm; IR (KBr) v max 210, 276 nm; IR (KBr) v max 3370, 2925, 2854, 1733, 1712, 1455, 1437, 1279, 1197, 734 cm –1 ; HRESIMS m / z 293.1759 [M-H] - (calcd for C 17 H 25 O4: 293.1753).

[0119] Compound 7. Toxicodenol G

[0120]

[0121] Yellow oil; UV (MeOH) λmax212, 360 nm; IR (KBr) v max 212, 360 nm; IR (KBr) v max 3369, 2924, 2852, 1703, 1475, 1453, 1279, 1195, 734, 697 cm –1 ; HRESIMS m / z 297.1855 [M-H] - (calcd for C 20 H 25 O2: 297.1855).

[0122] Compound 11. Toxicodenol K

[0123]

[0124] Yellow oil; UV (MeOH) λmax208, 240, 272 nm; IR (KBr) v max 208, 240, 272 nm; IR (KBr) v max 3369, 2925, 2853, 1682, 1595, 1475, 1280, 1204, 734, 690 cm –1 ; HRESIMS m / z 339.1963 [M-H] - (calcd for C 22 H 27 O3: 339.1960).

[0125] Compound 13. Toxicodenol M

[0126]

[0127] Yellow oil; UV (MeOH) λmax max 208, 230, 272 nm; IR (iTR) v max 3379, 2922, 2852, 1588, 1455, 1270, 1154, 779, 693 cm –1 ; HRESIMS m / z 357.3155 [M-H] - (calcd for C 25 H 41 O: 357.3157); EI-MS major fragments m / z 108.14, 261.30, 315.35.

[0128] Compound 15. Toxicodenol O

[0129]

[0130] Yellow oil; UV (MeOH) λmax max 216, 248, 304 nm; IR (iTR) v max 3353, 2926, 2854, 2158, 2024, 1683, 1585, 1451, 1279 cm –1 ; HRESIMS m / z 315.2326 [M-H] - (calcd for C 21 H 31 O2: 315.2324); HREI-MS major fragments m / z 136.05191, 217.12241; EI-MS major fragments m / z 85.18, 121.16, 136.20, 191.32.

[0131] Compound 17. Toxicodenol Q

[0132]

[0133] Yellow oil; UV (MeOH) λmax max 206, 240, 278, 320 nm; IR (iTR) v max 3396, 2926, 2853, 1682, 1597, 1587, 1449, 1270, 1216, 1178, 1155, 781, 751, 691 cm –1 ; HRESIMS m / z 309.1860 [M-H] - (calcd for C 21 H25 O2: 309.1855).

[0134] Compound 22. Toxicodenol V

[0135]

[0136] Yellow oil; UV (MeOH) λ max 206, 275, 340 nm; IR (iTR) v max 2922, 2852, 1737, 1638, 1433, 1317, 1277, 1232, 875, 786 cm –1 ; HRESIMS m / z 401.3054 [M-H] - (calcd for C 26 H 41 O3: 401.3056); HREI-MS major fragments m / z 57.03352, 180.07797, 345.27847; EI-MS major fragments m / z 57.07, 83.10, 180.12, 291.27, 345.35, 373.34, 388.33.

[0137] Compound 23. Toxicodenol W

[0138]

[0139] Yellow oil; UV (MeOH) λ max 212, 276, 342 nm; IR (iTR) v max 2920, 2850, 1737, 1463, 1178, 719 cm –1 ; HRESIMS m / z 389.3054 [M-H] - (calcd for C 25 H 41 O3: 389.3056).

[0140] Compound 24. 3-Hexylbenzene-1,2-diol

[0141]

[0142] Yellow oil; 1 HNMR (500 MHz, CDC13): δ H6.73 (3H, overlapped, H-4,5,6), 2.60 (2H, t, J = 7.9 Hz, H-1'), 1.61 (2H, m, H-2'), 1.26-1.38 (6H, m, H-3',4',5'), 0.89 (3H, t, J = 6.8 Hz, H-6'); 13 CNMR (125 MHz, CDC13): δ C 143.1 (C-1), 141.9 (C-2), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 31.8 (C-1'), 29.9 (C-4'), 29.8 (C-2'), 29.4 (C-3'), 22.8 (C-5'), 14.3 (C-6'); (-)-ESI-MS m / z 193 [M-H] - .

[0143] Compound 25, 3-Heptylbenzene-1,2-diol

[0144]

[0145] yellow oil; 1 HNMR (500 MHz, CDC13): δ H 6.71 (3H, overlapped, H-4,5,6), 2.60 (2H, t, J = 8.0 Hz, H-1'), 1.61 (2H, m, H-2'), 1.26-1.38 (8H, m, H-3',4',5',6'), 0.88 (3H, t, J = 7.3 Hz, H-7'); 13 CNMR (125 MHz, CDC13): δ C 143.0 (C-1), 141.9 (C-2), 129.4 (C-3), 122.1 (C-4), 120.1 (C-5), 112.9 (C-6), 31.8 (C-5'), 29.9 (C-2'), 29.8 (C-1'), 29.7 (C-3'), 29.5 (C-4'), 22.2 (C-6'), 14.1 (C-7'); (-)-ESI-MS m / z 207 [M-H] - .

[0146] Compound 26, 3-Octylbenzene-1,2-diol

[0147]

[0148] yellow oil; 1HNMR (500 MHz, CDC13): δ 6.71 (3H, overlapped, H-4,5,6), 2.60 (2H, t, J = 7.9 Hz, H-1'), 1.61 (2H, m, H-2'), 1.24-1.37 (10H, m, H-3',4',5',6',7'), 0.88 (3H, t, J = 6.8 Hz, H-8'); H 6.71 (3H, overlapped, H-4,5,6), 2.60 (2H, t, J = 7.9 Hz, H-1'), 1.61 (2H, m, H-2'), 1.24-1.37 (10H, m, H-3',4',5',6',7'), 0.88 (3H, t, J = 6.8 Hz, H-8'); 13 CNMR (125 MHz, CDC13): δ 143.2 (C-1), 142.0 (C-2), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 32.0 (C-6'), 29.9 (C-1'), 29.9 (C-4'), 29.9 (C-5'), 29.8 (C-2'), 29.7 (C-3'), 22.8 (C-7'), 14.3 (C-8'); C 143.2 (C-1), 142.0 (C-2), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 32.0 (C-7'), 29.9 (C-1'), 29.9 (C-6'), 29.8 (C-2'), 29.8 (C-5'), 29.7 (C-3'), 29.7 (C-4'), 22.8 (C-8'), 14.3 (C-9'); (-)-ESI-MS m / z 235 [M-H] - .

[0149] Compound 27, 3-Nonylbenzene-1,2-diol

[0150]

[0151] yellow oil; 1 HNMR (500 MHz, CDC13): δ 6.71 (3H, overlapped, H-4,5,6), 2.60 (2H, t, J = 7.9 Hz, H-1'), 1.61 (2H, m, H-2'), 1.24-1.37 (10H, m, H-3',4',5',6',7'), 0.88 (3H, t, J = 6.8 Hz, H-8'); H 6.71 (3H, overlapped, H-4,5,6), 2.60 (2H, t, J = 7.9 Hz, H-1'), 1.61 (2H, m, H-2'), 1.24-1.37 (10H, m, H-3',4',5',6',7'), 0.88 (3H, t, J = 6.8 Hz, H-8'); 13 CNMR (125 MHz, CDC13): δ 143.2 (C-1), 142.0 (C-2), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 32.0 (C-7'), 29.9 (C-1'), 29.9 (C-6'), 29.8 (C-2'), 29.8 (C-5'), 29.7 (C-3'), 29.7 (C-4'), 22.8 (C-8'), 14.3 (C-9'); C 143.2 (C-1), 142.0 (C-2), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 32.0 (C-7'), 29.9 (C-1'), 29.9 (C-6'), 29.8 (C-2'), 29.8 (C-5'), 29.7 (C-3'), 29.7 (C-4'), 22.8 (C-8'), 14.3 (C-9'); (-)-ESI-MS m / z 235 [M-H] - .

[0152] Compound 28, 3-Decylbenzene-1,2-diol

[0153]

[0154] Yellow oil; 1 HNMR (500 MHz, CDC13): δ H 6.72 (3H, overlapped, H-4,5,6), 2.61 (2H, t, J = 7.8 Hz, H-1'), 1.62 (2H, m, H-2'), 1.27-1.38 (14H, m, H-3',4',5',6',7',8',9'), 0.88 (3H, t, J = 6.9 Hz, H-10'); 13 CNMR (125 MHz, CDC13): δ C 143.2 (C-1), 142.0 (C-2), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 32.1 (C-8'), 29.9 (C-1'), 29.9 (C-7'), 29.8 (C-2'), 29.8 (C-3'), 29.8 (C-5'), 29.8 (C-6'), 29.7 (C-4'), 22.8 (C-9'), 14.3 (C-10'); (-)-ESI-MS m / z 249 [M-H] - .

[0155] Compound 29, 3-Undecylbenzene-1,2-diol

[0156]

[0157] Yellow oil; 1 HNMR (500 MHz, CDC13): δ H 6.72 (3H, overlapped, H-4,5,6), 2.61 (2H, t, J = 7.8 Hz, H-1'), 1.62 (2H, m, H-2'), 1.27-1.38 (14H, m, H-3',4',5',6',7',8',9'), 0.88 (3H, t, J = 6.9 Hz, H-10'); 13 CNMR (125 MHz, CDC13): δ C143.2 (C-1), 142.0 (C-2), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 32.1 (C-9'), 29.9 (C-1'), 29.9 (C-8'), 29.8 (C-2'), 29.8 (C-3'), 29.8 (C-6'), 29.8 (C-7'), 29.7 (C-4'), 29.7 (C-5'), 22.8 (C-10'), 14.3 (C-11'); (-)-ESI-MS m / z 263 [M-H] - .

[0158] Compound 30, (Z)-3-(Pentadec-8-enyl)benzene-1,2-diol

[0159]

[0160] yellow oil; 1 HNMR (500 MHz, CDC13): δ H 6.73 (3H, overlapped, H-4,5,6), 5.37 (2H, m, H-8',9'), 2.61 (2H, t, J = 7.8 Hz, H-1'), 2.03 (4H, q, J = 6.4 Hz, H-7',10'), 1.62 (2H, m, H-2'), 1.27-1.38 (16H, m, H-3',4',5',6',11',12',13',14'), 0.90 (3H, t, J = 6.7 Hz, H-15'); 13 CNMR (125 MHz, CDC13): δ C 143.1 (C-1), 142.0 (C-2), 130.1 (C-8'), 129.9 (C-9'), 129.5 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 31.9 (C-13'), 29.9 (C-1'), 29.9 (C-2'), 29.8 (C-3'), 29.8 (C-11'), 29.8 (C-12'), 29.7 (C-4'), 29.7 (C-5'), 29.5 (C-6'), 27.6 (C-7'), 27.7 (C-10'), 22.8 (C-14'), 14.3 (C-15'); (-)-ESI-MS m / z 317 [M-H] - ; EI-MS major fragments m / z 85.95, 111.14, 123.08, 233.22.

[0161] Compound 31, 3-Heptadecylbenzene-1,2-dio

[0162]

[0163] yellow oil; 1 HNMR (500 MHz, CDC13): δ H 6.72 (3H, overlapped, H-4,5,6), 2.59 (2H, t, J = 7.8 Hz, H-1'), 1.59 (2H, m, H-2'), 1.26-1.35 (24H, m, H-3',4',5',6',7',8',9',10',11',12',13',14'), 0.89 (3H, t, J = 6.8 Hz, H-17'); 13 CNMR (125 MHz, CDC13): δ C 144.9 (C-1), 142.9 (C-2), 128.5 (C-3), 123.2 (C-4), 122.2 (C-5), 114.0 (C-6), 31.9 (C-15'), 29.9 (C-1'), 29.9 (C-13'), 29.8 (C-2'), 29.8 (C-4'), 29.8 (C-5'), 29.8 (C-6'), 29.7 (C-7'), 29.7 (C-8'), 29.7 (C-9'), 29.7 (C-10'), 29.6 (C-11'), 29.6 (C-12'), 29.4 (C-3'), 29.3 (C-14'), 22.7 (C-16'), 14.3 (C-17'); (-)-ESI-MS m / z 347 [M-H] - .

[0164] Compound 32, (Z)-3-(Heptadec-10-enyl)benzene-1,2-diol

[0165]

[0166] yellow oil; 1 HNMR (500 MHz, CDC13): δ H6.71 (3H, overlapped, H-4,5,6), 5.36 (2H, m, H-10',11'), 2.60 (2H, t, J = 7.9 Hz, H-1'), 2.02 (4H, q, J = 6.5 Hz, H-9',12'), 1.62 (2H, m, H-2'), 1.28-1.38 (16H, m, H-3',4',5',6',7',8',13',14',15',16'), 0.89 (3H, t, J = 6.8 Hz, H-17'); 13 CNMR (125 MHz, CDC13): δ C 143.2 (C-1), 142.0 (C-2), 130.0 (C-8'), 130.0 (C-9'), 129.9 (C-3), 122.2 (C-4), 120.2 (C-5), 113.0 (C-6), 31.9 (C-15'), 29.9 (C-1'), 29.9 (C-2'), 29.9 (C-3'), 29.8 (C-8'), 29.8 (C-11'), 29.8 (C-12'), 29.7 (C-4'), 29.7 (C-5'), 29.5 (C-6'), 29.5 (C-7'), 27.6 (C-9'), 27.7 (C-12'), 22.8 (C-16'), 14.3 (C-17'); (-)-ESI-MS m / z 345 [M-H] - ; EI-MS major fragments m / z 85.99, 111.12, 123.10, 261.25.

[0167] Compound 33, (E)-3-(Heptadec-10-enyl)benzene-1,2-diol

[0168]

[0169] yellow oil; 1 HNMR (500 MHz, CDC13): δ H 6.69 (3H, overlapped, H-4,5,6), 5.38 (2H, m, H-10',11'), 2.59 (2H, t, J = 7.9 Hz, H-1'), 1.97 (4H, q, J = 6.5 Hz, H-9',12'), 1.58 (2H, m, H-2'), 1.25-1.36 (16H, m, H-3',4',5',6',7',8',13',14',15',16'), 0.88 (3H, t, J = 6.8 Hz, H-17'); 13 CNMR (125 MHz, CDC13): δC 143.3 (C-1), 142.2 (C-2), 130.5 (C-8'), 130.1 (C-9'), 129.5 (C-3), 122.0 (C-4), 120.0 (C-5), 112.8 (C-6), 32.7 (C-9'), 31.9 (C-12'), 29.9 (C-15'), 29.9 (C-1'), 29.9 (C-2'), 29.9 (C-3'), 29.8 (C-8'), 29.8 (C-11'), 29.8 (C-12'), 29.7 (C-4'), 29.7 (C-5'), 29.5 (C-6'), 29.5 (C-7'), 22.8 (C-16'), 14.2 (C-17'); (-)-ESI-MS m / z 345 [M-H] - ; EI-MS major fragments m / z 85.97, 111.14, 123.08, 275.33.

[0170] Compound 34. 3-Nonylphenol

[0171]

[0172] yellow oil; 1 HNMR (500 MHz, CDC13): δ H 7.14 (1H, t, J = 7.7 Hz, H-5), 6.77 (1H, d, J = 7.7 Hz, H-6), 6.67 (1H, d, J = 7.7 Hz, H-4), 6.65 (1H, s, H-2), 2.56 (2H, t, J = 7.7 Hz, H-1'), 1.61 (2H, m, H-2'), 1.27-1.34 (12H, m, H-3', 4', 5', 6', 7', 8'), 0.88 (3H, t, J = 6.8 Hz, H-9'); 13 CNMR (125 MHz, CDC13): δ C 155.4 (C-1), 144.9 (C-3), 129.4 (C-5), 120.9 (C-4), 115.3 (C-6), 112.5 (C-2), 35.8 (C-1'), 31.9 (C-2'), 31.3 (C-7'), 29.7 (C-3'), 29.6 (C-4'), 29.6 (C-5'), 29.5 (C-6'), 22.7 (C-8'), 14.2 (C-9'); (-)-ESI-MS m / z 219 [M-H] - .

[0173] Compound 35. 3-Pentadecylphenol

[0174]

[0175] Yellow oil; 1 HNMR (500 MHz, CDC13): δ H 7.14 (1H, t, J = 7.7 Hz, H-5), 6.76 (1H, d, J = 7.7 Hz, H-6), 6.66 (1H, d, J = 7.7 Hz, H-4), 6.64 (1H, s, H-2), 2.55 (2H, t, J = 7.8 Hz, H-1'), 1.98 (2H, m, H-2'), 1.25-1.32 (24H, m, H-3',4',5',6',7',8',9',10',11',12',13',14'), 0.88 (3H, t, J = 6.7 Hz, H-15'); 13 CNMR (125 MHz, CDC13): δ C 156.4 (C-1), 143.7 (C-3), 129.9 (C-5), 120.6 (C-4), 113.2 (C-6), 114.5 (C-2), 35.9 (C-1'), 31.7 (C-13'), 30.9 (C-2'), 29.9 (C-3'), 29.9 (C-4'), 29.9 (C-12'), 29.9 (C-5'), 29.8 (C-6'), 29.8 (C-11'), 29.7 (C-7'), 29.7 (C-8'), 29.6 (C-9'), 29.6 (C-10'), 22.7 (C-14'), 14.2 (C-15'); (-)-ESI-MS m / z 303 [M-H] - .

[0176] Compound 36. (E)-3-(Pentadec-8-enyl)phenol

[0177]

[0178] Yellow oil; 1 HNMR (500 MHz, CDC13): δ H7.15 (1H, t, J = 7.6 Hz, H-5), 6.76 (1H, d, J = 7.6 Hz, H-6), 6.66 (1H, d, J = 7.6 Hz, H-4), 6.64 (1H, s, H-2), 5.36 (2H, m, H-8', 9'), 2.56 (2H, t, J = 7.8 Hz, H-1'), 2.02 (4H, q, J = 6.4 Hz, H-7', 10'), 1.59 (2H, m, H-2'), 1.26-1.35 (16H, m, H-3', 4', 5', 6', 11', 12', 13', 14'), 0.89 (3H, t, J = 6.7 Hz, H-15'); 13 CNMR (125 MHz, CDC13): δ C 155.5 (C-1), 145.1 (C-3), 130.0 (C-8'), 129.9 (C-9'), 129.5 (C-5), 121.1 (C-4), 115.4 (C-6), 112.6 (C-2), 35.9 (C-1'), 33.7 (C-7'), 33.6 (C-10'), 31.9 (C-13'), 31.4 (C-2'), 29.9 (C-3'), 29.9 (C-4'), 29.9 (C-12'), 29.8 (C-5'), 29.8 (C-6'), 29.8 (C-11'), 22.7 (C-14'), 14.2 (C-15'); (-)-ESI-MS m / z 301 [M-H] - ; EI-MS major fragments m / z 71.10, 85.96, 108.11, 125.14.

[0179] Compound 37. (Z)-3-(Heptadec-8-enyl)phenol

[0180]

[0181] yellow oil; 1 HNMR (500 MHz, CDC13): δ H7.15 (1H, t, J = 7.7 Hz, H-5), 6.76 (1H, d, J = 7.7 Hz, H-6), 6.66 (1H, d, J = 7.7 Hz, H-4), 6.64 (1H, s, H-2), 5.39 (2H, m, H-8', 9'), 2.56 (2H, t, J = 7.8 Hz, H-1'), 1.97 (4H, q, J = 6.2 Hz, H-7', 10'), 1.60 (2H, m, H-2'), 1.26-1.34 (20H, m, H-3', 4', 5', 6', 11', 12', 13', 14', 15', 16'), 0.89 (3H, t, J = 6.8 Hz, H-17'); 13 CNMR (125 MHz, CDC13): δ C 155.5 (C-1), 144.9 (C-3), 130.4 (C-8'), 130.3 (C-9'), 129.4 (C-5), 120.9 (C-4), 115.3 (C-6), 112.4 (C-2), 35.8 (C-1'), 32.6 (C-15'), 31.5 (C-2'), 29.9 (C-3'), 29.9 (C-4'), 29.9 (C-5'), 29.8 (C-13'), 29.8 (C-12'), 29.8 (C-6'), 29.7 (C-11'), 29.7 (C-14'), 27.7 (C-7'), 27.6 (C-10'), 22.7 (C-16'), 14.2 (C-17'); (-)-ESI-MS m / z 329 [M-H] - ; EI-MS major fragments m / z 57.09, 108.10, 149.16, 177.08, 217.12, 273.30.

[0182] Compound 38. 8-(3-Hydroxyphenyl)octanoic acid

[0183]

[0184] yellow oil; 1 HNMR (500 MHz, CDC13): δ H7.12 (1H, t, J = 7.6 Hz, H-5), 6.73 (1H, d, J = 7.6 Hz, H-6), 6.66 (1H, d, J = 7.6 Hz, H-4), 6.63 (1H, s, H-2), 2.54 (2H, t, J = 7.7 Hz, H-1'), 2.35 (2H, t, J = 7.5 Hz, H-7'), 1.61 (2H, m, H-2'), 1.57 (2H, m, H-6'), 1.29-1.32 (6H, m, H-3', 4', 5'); 13 CNMR (125 MHz, CDC13): δ C 179.9 (C-8'), 155.5 (C-1), 144.7 (C-3), 129.4 (C-5), 120.8 (C-4), 115.3 (C-6), 113.1 (C-2), 35.7 (C-1'), 34.1 (C-7'), 31.3 (C-2'), 29.9 (C-3'), 29.8 (C-4'), 29.7 (C-5'), 24.6 (C-6'); (-)-ESI-MS m / z 235 [M-H] - .

[0185] Compound 39. 9-(3-Hydroxyphenyl)nonanoic acid

[0186]

[0187] yellow oil; 1 HNMR (500 MHz, CDC13): δ H 7.12 (1H, t, J = 7.6 Hz, H-5), 6.73 (1H, d, J = 7.6 Hz, H-6), 6.66 (1H, d, J = 7.6 Hz, H-4), 6.63 (1H, s, H-2), 2.54 (2H, t, J = 7.7 Hz, H-1'), 2.35 (2H, t, J = 7.5 Hz, H-7'), 1.61 (2H, m, H-2'), 1.57 (2H, m, H-6'), 1.29-1.32 (6H, m, H-3', 4', 5'); 13 CNMR (125 MHz, CDC13): δ C179.8 (C-9'), 155.6 (C-1), 144.9 (C-3), 129.5 (C-5), 120.9 (C-4), 115.5 (C-6), 112.6 (C-2), 35.8 (C-1'), 34.2 (C-8'), 31.3 (C-2'), 29.4 (C-3'), 29.3 (C-4'), 29.3 (C-5'), 29.2 (C-5'), 24.8 (C-7'); (-)-ESI-MS m / z 249 [M-H] - .

[0188] Example 2, TGF-β-induced cardiac fibrosis activity screening

[0189] 1. Isolation and treatment of cardiac fibroblasts

[0190] Cardiac fibroblasts were isolated from neonatal SD rats using mixed enzyme digestion (0.06% trypsin and 0.04% collagenase type II dissolved in D-PBS without Ca 2+ and Mg 2+ ). The 2-6th generation cells were cultured with serum-free medium for 24 h, and then stimulated with 20 ng / mL TGF-β1 (PeproTech, USA, batch number: 0616354) for 48 h by differential adhesion of fibroblasts and cardiomyocytes for 90 min. The cells and supernatant were collected for further experiments. Finally, the positive drug tanshinone IIA and the test compound were diluted with DMSO, added with 10% fetal bovine serum, 1% penicillin and streptomycin, and obtained serial solutions of different concentrations (10, 20, 40 μM) for subsequent experiments.

[0191] 2. CCK-8 detection of cell relative viability

[0192] The 2-3th generation cardiac fibroblasts in the logarithmic growth phase were cultured in a 96-well plate at a density of 10 4 cells / well, and the cells were cultured in a 37°C constant temperature and 5% CO2 saturated incubator for 24 h using complete medium containing 10% fetal bovine serum. The serum was taken overnight to make the cells in the stationary growth phase, and then divided into a control group, a TGF-β (20 ng / mL -1 ) group, and a TGF-β (20 ng / mL -1)+ compound (10, 20, 40 μM) administration group, 5 replicates in each group, intervention time 48 h. Then discard the culture medium, add 100 μL culture medium and 10 μL CCK-8 solution to each well in the dark. Put the 96-well plate into a 37℃, 5% CO2 incubator for 4 h, then measure the OD value at 450 nm with a microplate reader. The cell proliferation rate is calculated according to the following formula: cell proliferation rate = average value of OD value of each intervention group cell / average value of OD value of control group cell. Take the control group as 100% reference value, compare the values of other groups to get the relative proliferation rate.

[0193] The in vitro myocardial fibrosis model was constructed by stimulating neonatal SD rat myocardial fibroblasts with 20 ng / mL TGF-β1 for 48 h, and the relative cell viability was detected by CCK-8, with tanshinone IIA as a positive drug. The results showed that compounds 1*, 2*, 5*, 6*, 8*, 10*, 12*, 14*, 19*, 20*, 21*, 29, 30, 33 and 37 had good inhibitory activity on myocardial fibrosis and showed concentration dependence; compound 35 had poor concentration dependence on inhibitory activity of myocardial fibrosis; compounds 9*, 16* and 18* had high inhibitory activity at low concentration of 10 μM. Figure 24

[0194] Obviously, those skilled in the art can make various modifications and variations to the present application without departing from the spirit and scope of the present application. Thus, if these modifications and variations of the present application fall within the scope of the claims of the present application and their equivalent technologies, the present application also intends to include these modifications and variations.​

Claims

1. A urushibara derivatives as follows: ###0001### 。 2. Use of the urushibara derivatives of claim 1 for the manufacture of a medicament for the prevention of myocardial fibrosis.

3. A medicament for the prevention of myocardial fibrosis, said medicament comprising at least one of the urushibara derivatives of claim 1.

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