A furanocoumarin compound, a preparation method and application thereof

Abstract

The application belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to a furanocoumarin compound, a preparation method and application thereof. The furanocoumarin compound provided by the application has a structure shown in formula I. The furanocoumarin compound provided by the application can inhibit the release of inflammatory factors, has an antioxidation effect, can inhibit the NF-kappa B and MAPK signal pathways, thereby playing an anti-inflammatory role, and is used for preparing anti-inflammatory drugs. Moreover, the furanocoumarin compound is extracted from radix angelicae dahuricae, is natural in origin, and is simple and efficient in the preparation method. Formula I.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine extraction technology, specifically relating to a furanocoumarin compound, its preparation method and application. Background Technology

[0002] Angelica dahurica is the dried root of Angelica dahurica or Angelica dahurica var. hainanensis, both belonging to the Apiaceae family. It is warm in nature and pungent in taste, and enters the stomach, large intestine, and lung meridians. Angelica dahurica has the effects of dispelling wind and dampness, clearing the orifices and relieving pain, reducing swelling and draining pus. It is used to treat colds with headaches, supraorbital neuralgia, nasal congestion, sinusitis, toothache, leukorrhea, and sores and swellings. The chemical components of Angelica dahurica include coumarins and volatile oils, among which the main coumarins are imperatorin, eugenol, and isoeugenol. Current research mainly focuses on coumarins, and no reports have been found on furanocoumarin compounds. Summary of the Invention

[0003] In view of this, the purpose of this invention is to provide a furanocoumarin compound, its preparation method and application. The furanocoumarin compound extracted from Angelica dahurica by this invention has good anti-inflammatory effects.

[0004] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a furanocoumarin compound having the structure shown in Formula I: Formula I.

[0005] This invention also provides a method for preparing the furanocoumarin compounds described in the above technical solution, comprising the following steps: Angelica root powder and extraction solvent were mixed and subjected to ultrasonic extraction. The resulting extract was concentrated under reduced pressure to obtain an extract. The extract was initially separated by a macroporous resin column, and the resulting active substances were then separated by silica gel column chromatography and semi-preparative liquid chromatography to obtain the furanocoumarin compounds.

[0006] Preferably, the particle size of the Angelica dahurica root powder is 10-60 mesh.

[0007] Preferably, the extraction solvent includes an organic alcohol or ethyl acetate; the organic alcohol includes one or more of ethanol, methanol and propanol.

[0008] Preferably, the ultrasonic extraction power is 200~400W, the frequency is 20~30Hz, the temperature is 50~70℃, and the time is 30~60min.

[0009] Preferably, the pressure for vacuum concentration is 10~30 mmHg, the temperature is 35~45℃, and the time is 40~80 min.

[0010] Preferably, the resin in the macroporous resin column is AB-8 resin.

[0011] Preferably, the eluent used in the silica gel column chromatography includes petroleum ether and ethyl acetate, or petroleum ether and dichloromethane, or dichloromethane and methanol.

[0012] Preferably, the conditions for the semi-preparative liquid chromatography separation include: Chromatographic column: C18 column, dimensions: 250mm × 10mm, packing material particle size: 5~10µm. The mobile phase composition by volume ratio is: acetonitrile:water = 55~60:40~45. Flow rate: 1~2 mL / min Column temperature: 20~25℃ Detection wavelength: 254nm Injection volume: 500~1500µL Concentration of the injection solution: 0.5~2 mg / mL.

[0013] The present invention also provides the application of the furanocoumarin compounds described in the above technical solutions or the furanocoumarin compounds prepared by the preparation methods described in the above technical solutions in the preparation of anti-inflammatory drugs.

[0014] This invention provides a furanocoumarin compound having the structure shown in Formula I: Formula I.

[0015] The furanocoumarin compounds provided by this invention can inhibit the release of inflammatory factors, have antioxidant effects, and also inhibit the NF-κB and MAPK signaling pathways, thereby exerting anti-inflammatory effects and can be used to prepare anti-inflammatory drugs. Moreover, the furanocoumarin compounds are extracted from Angelica dahurica, making them a natural source, and the preparation method is simple and efficient. Attached Figure Description

[0016] Figure 1 The effects of compounds in Angelica dahurica on the secretion levels of NO, TNF-α, IL-6 and IL-1β in LPS-induced RAW 264.7 macrophages; Figure 2 The effect of furanocoumarin compounds in Angelica dahurica on the viability of LPS-induced RAW 264.7 macrophages; Figure 3 The effects of compounds 2 and 3 on the LPS-induced NF-κB signaling pathway in RAW264.7 cells; Figure 4 The effects of compounds 2 and 3 on the MAPK signaling pathway in LPS-induced RAW264.7 cells. Detailed Implementation

[0017] This invention provides a furanocoumarin compound having the structure shown in Formula I: Formula I.

[0018] As one embodiment, the furanocoumarin compound having the structure shown in Formula I is named 2-(Z)-butenoic-2-methyl-2-[(8S)-8,9-dihydro-2-oxo-2H-furo[2,3-h]chromen-8-yl]allylester, with the molecular formula: C 19 H 18 O5, precise molecular weight: 326.1154, molar mass: 326.3480, appearance: white amorphous powder.

[0019] The furanocoumarins in Angelica dahurica have anti-inflammatory effects, mainly because these compounds possess certain biological activities and can exert their anti-inflammatory effects through multiple mechanisms. Specific reasons include: 1. Inhibition of inflammatory cytokine release: Furanocoumarins can inhibit the release of some key inflammatory cytokines, such as tumor necrosis factor (TNF-α) and interleukins (IL-1β, IL-6), which play important roles in the inflammatory response. By inhibiting the synthesis or release of these cytokines, the inflammatory response can be reduced.

[0020] 2. Antioxidant effect: Furanocoumarins have strong antioxidant capabilities, can scavenge free radicals, and reduce oxidative stress. Oxidative stress is an important factor leading to chronic inflammation and tissue damage, and antioxidant effects can effectively reduce inflammatory responses.

[0021] 3. Inhibition of NF-κB and MAPK signaling pathways: By inhibiting these signaling pathways, furanocoumarins can reduce the expression of inflammation-related genes and reduce inflammatory responses.

[0022] This invention also provides a method for preparing the furanocoumarin compounds described in the above technical solution, comprising the following steps: Angelica root powder and extraction solvent were mixed and subjected to ultrasonic extraction. The resulting extract was concentrated under reduced pressure to obtain an extract. The extract was initially separated by a macroporous resin column, and the resulting active substances were then separated by silica gel column chromatography and semi-preparative liquid chromatography to obtain the furanocoumarin compounds.

[0023] Unless otherwise specified, the present invention does not have special requirements on the source of raw materials used, and commercially available products well known to those skilled in the art can be used.

[0024] In one embodiment, the particle size of the Angelica dahurica root powder is 10-60 mesh, and in a specific embodiment, it is 40-60 mesh. This particle size range can increase the extraction contact area, improve the permeability of organic solvents, and promote the release of furanocoumarin compounds, thereby improving the extraction efficiency.

[0025] As one implementation method, before mixing the Angelica dahurica root powder and the extraction solvent, the method further includes: drying the Angelica dahurica root powder to a constant weight; the drying is sun drying; the constant weight is defined as the difference between two consecutive weighings being less than 0.1g; the moisture content of the dried Angelica dahurica root powder is ≤10%, and in a specific embodiment it is ≤5%.

[0026] In one embodiment, the extraction solvent includes an organic alcohol or ethyl acetate; the organic alcohol includes one or more of ethanol, methanol, and propanol, with ethanol being used in a specific embodiment; the ethanol is used in the form of a 70% (v / v) aqueous solution. Compared to other extraction solvents, ethanol has better solubility and can extract compounds of moderate polarity; at the same time, ethanol has low toxicity and high safety, making it suitable for the extraction of medicinal plants; its volatility is moderate, facilitating concentration after extraction; furthermore, ethanol has a wide range of applications in plant extraction with abundant experimental support, thus making it an ideal solvent for extracting furanocoumarin compounds from Angelica dahurica.

[0027] In one embodiment, the mass ratio of the Angelica dahurica root powder to the extraction solvent is 1:10~20, and in a specific embodiment it is 1:15.

[0028] In one implementation method, the ultrasonic extraction power is 200-400W, specifically 200-300W in this embodiment; the frequency is 20-30kHz, specifically 20-25kHz in this embodiment; the temperature is 50-70℃, specifically 50-60℃ in this embodiment; and the time is 30-60min, specifically 30-40min in this embodiment. Ultrasonic extraction accelerates the dissolution process through mechanical vibration and cavitation effects, disrupts plant cell walls, and increases the permeability of the extraction solvent, thereby improving the extraction efficiency of furanocoumarin compounds. Ultrasonic extraction can also shorten the extraction time, reduce the impact of high temperatures on heat-sensitive components, and maintain the stability of the compounds, making it an effective method for extracting furanocoumarin compounds from Angelica dahurica.

[0029] In one implementation method, the ultrasonic extraction is repeated 2 to 4 times, and in a specific embodiment, it is repeated 3 times.

[0030] In one implementation method, the pressure for vacuum concentration is 10-30 mmHg, specifically 20 mmHg in this embodiment; the temperature is 35-45°C, specifically 40°C in this embodiment; and the time is 40-80 min, specifically 50-60 min in this embodiment. This invention, through vacuum concentration, allows the extraction solvent to evaporate at a lower temperature, reducing the degradation of heat-sensitive components, and ultimately concentrating the solution to approximately 10-20% of its original volume, yielding a viscous extract. This invention ensures that the final extract reaches the ideal concentration or viscosity through vacuum concentration, facilitating subsequent processing.

[0031] In one embodiment, the extract is initially separated by passing it through a macroporous resin column: The extract was dissolved in water, and the resulting solution was passed through a macroporous resin column for adsorption. After adsorption equilibrium was reached, the macroporous resin column was eluted with an eluent. The resulting eluent was concentrated under reduced pressure and dried to obtain the active substance.

[0032] In one embodiment, the concentration of the loading solution is 5-20 mg / mL, specifically 10 mg / mL in this embodiment; the loading volume is 50-200 mL, specifically 100 mL in this embodiment; the loading flow rate is 1-3 BV / h, specifically 2 BV / h in this embodiment; the resin in the macroporous resin column is AB-8 resin; the eluent is an ethanol solution; the volume concentration of the ethanol solution is 30%; the elution time is 30 min-2 h, specifically 1 h in this embodiment; the eluent flow rate is 1-5 mL / min, specifically 3 mL / min in this embodiment; the pressure for vacuum concentration is 10-30 mmHg, specifically 20 mmHg in this embodiment; the time is 40-80 min, specifically 50-60 min in this embodiment; the drying temperature is 35-45℃, specifically 40℃ in this embodiment; the drying time is 2-12 h, specifically 6-8 h in this embodiment.

[0033] In one embodiment, the silica gel column used for the silica gel column chromatography is a glass column packed with 200-300 mesh silica gel; the eluent used for the silica gel column chromatography includes petroleum ether and ethyl acetate, or petroleum ether and dichloromethane, or dichloromethane and methanol, with petroleum ether and ethyl acetate being used in a specific embodiment; the volume ratio of petroleum ether to ethyl acetate is 1-5:1; the specific steps of the silica gel column chromatography are as follows: the active substance is dissolved in dichloromethane or ethyl acetate to obtain an active substance solution; the active substance solution is adsorbed onto the silica gel column, and the adsorbed silica gel column is eluted with the eluent used for silica gel column chromatography; the resulting eluent is collected, the same components are combined, and concentrated under reduced pressure; the concentration of the active substance solution is 50-200 mg / mL, with a specific embodiment... The concentration is 100 mg / mL; the elution time is 2-4 h, specifically 3 h in this embodiment; the elution procedure is as follows: elute sequentially with a petroleum ether and ethyl acetate eluent at a volume ratio of 5:1 for 30-60 min, elute with a petroleum ether and ethyl acetate eluent at a volume ratio of 3:1 for 30-60 min, and elute with a petroleum ether and ethyl acetate eluent at a volume ratio of 1:1 for 60-120 min until the target compound is completely eluted; the flow rate of the eluent used in the silica gel column chromatography is 2-5 mL / min, specifically 3 mL / min in this embodiment; the pressure for the reduced pressure concentration is 10-30 mmHg, specifically 20 mmHg in this embodiment, and the time is 40-80 min, specifically 50-60 min in this embodiment.

[0034] As one embodiment, the specific steps of the semi-preparative liquid chromatography separation are as follows: dissolving the substance obtained from silica gel column chromatography in methanol or acetonitrile to obtain an injection solution; injecting the injection solution into a semi-preparative liquid chromatograph, monitoring each fraction eluted from the chromatographic column, determining the position of the furanocoumarin compounds based on the retention time, and collecting the corresponding fractions through a fractionating distiller or an automatic collection system; the obtained fractions are concentrated under reduced pressure or freeze-dried to remove the solvent, and then the furanocoumarin compounds are obtained; the monitoring methods include ultraviolet detection, fluorescence detection, or mass spectrometry detection.

[0035] As one implementation method, the conditions for the semi-preparative liquid chromatography separation include: Chromatographic column: C18 column, dimensions: 250mm × 10mm, packing particle size: 5~10µm, 10µm in the specific embodiment. The mobile phase composition, by volume ratio, is acetonitrile:water = 55~60:40~45, with a specific ratio of 55:45 or 60:40 in the concrete examples. Flow rate: 1~2 mL / min, 2 mL / min in the specific example. Column temperature: 20~25℃, 22℃ in the specific embodiment. Detection wavelength: 254nm Injection volume: 500~1500µL, 1000µL in the specific example. The concentration of the injection solution is 0.5~2 mg / mL, and in the specific example it is 1 mg / mL.

[0036] In one embodiment, the pressure of the vacuum concentration is 10~30 mmHg, specifically 20 mmHg in this embodiment, and the time is 40~80 min, specifically 50~60 min in this embodiment; the temperature of the freeze drying is -40~-60℃, specifically -50℃ in this embodiment, and the time is 12~24 h, specifically 18~20 h in this embodiment.

[0037] In semi-preparative liquid chromatography (SLC), the process of identifying the target fraction typically involves several key steps. First, the mixture is separated using a liquid chromatography column (e.g., a C18 column), selecting an appropriate mobile phase (e.g., acetonitrile:water = 55:45) to ensure effective separation of the target compound from other components. Then, the target compound is identified in real-time based on changes in the detection signal by monitoring the individual fractions eluting from the column, typically using methods such as UV detection, fluorescence detection, or mass spectrometry. The identification of the target compound relies primarily on its retention time, i.e., the time required for the target substance to elute from the sample into the column; this characteristic can be used to accurately locate the target fraction. Finally, once the target fraction is identified, it is collected using a fractionator or an automated collection system. This invention, through parameter settings in semi-preparative liquid chromatography, helps optimize separation efficiency, and by monitoring retention time and detection signals, the fraction of the target compound can be accurately collected. The mobile phase used in this invention is suitable for separating moderately polar compounds, achieving a balance between good resolution and fluidity.

[0038] As one embodiment, the furanocoumarin compounds obtained by the preparation method of the furanocoumarin compounds have a purity of ≥95%, specifically 97%, 97.5% or 96.5% in the embodiments, and a yield of 0.005%~0.01%, specifically 0.008%, 0.0092% or 0.0095% in the embodiments (based on the weight of dried Angelica dahurica root powder).

[0039] The separation and extraction technology of this invention effectively separates and extracts furanocoumarin compounds from Angelica dahurica using a variety of techniques, including semi-preparative HPLC, reduced pressure concentration, and macroporous resin column chromatography, and has the following technical advantages: 1. High-efficiency separation: Semi-preparative HPLC (e.g., C18 column, acetonitrile:water = 55:45 mobile phase) can accurately separate complex chemical components in Angelica dahurica, especially furanocoumarins. The combination of acetonitrile and water provides good resolution, effectively separating the target compound from other impurities and ensuring the purity of the separation.

[0040] 2. Improved extraction efficiency: Elution with macroporous resin (such as AB-8 column) and 30 vol% ethanol not only helps remove nonpolar impurities but also effectively retains the target compound (furanocoumarins). Appropriate elution conditions ensure high recovery rates and high purity of furanocoumarin compounds.

[0041] 3. Avoiding thermal degradation: Reduced pressure concentration technology removes solvents at lower temperatures by reducing pressure and temperature, thus minimizing solvent evaporation and degradation of heat-sensitive components. This helps maintain the stability of furanocoumarins and prevents them from being destroyed at high temperatures.

[0042] 4. Simplified operation and reduced time: Optimized separation processes (such as combining semi-preparative HPLC with resin column chromatography) reduce the waste of time and solvents in traditional extraction methods, improving work efficiency. Simultaneously, using semi-preparative HPLC can reduce interference from other complex impurities, further enhancing the quality of the extract.

[0043] The present invention also provides the application of the furanocoumarin compounds described in the above technical solutions or the furanocoumarin compounds prepared by the preparation methods described in the above technical solutions in the preparation of anti-inflammatory drugs.

[0044] The present invention does not specifically limit the application method of the furanocoumarin compounds in the preparation of anti-inflammatory drugs; any application method known in the art can be used.

[0045] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, but they should not be construed as limiting the scope of protection of the present invention.

[0046] Example 1 Furanocoumarin compounds have the structure shown in Formula I: Formula I, its preparation method is as follows: Angelica root powder with a particle size of 40-60 mesh was dried to constant weight by sun drying (the difference between two consecutive weighings was less than 0.1g, and the moisture content was ≤5%). It was then mixed with a 70% ethanol aqueous solution at a mass ratio of 1:15 and subjected to repeated ultrasonic extraction 3 times. The ultrasonic extraction power was 300W, the frequency was 20kHz, the temperature was 60℃, and the ultrasonic extraction time was 30min each time. The resulting extract was concentrated under reduced pressure at 40℃ and 20mmHg for 1h to obtain the extract. The extract was dissolved in water, and the resulting column solution was adsorbed onto a macroporous resin column. After adsorption equilibrium was reached, the adsorbed macroporous resin column was eluted with an eluent. The resulting eluent was concentrated under reduced pressure at a flow rate of 3 mL / min and a pressure of 20 mmHg for 60 min, and then dried at 40 °C for 6 h to obtain the active substance. The concentration of the column solution was 10 mg / mL, the loading volume was 100 mL, the loading flow rate was 2 BV / h, the resin in the macroporous resin column was AB-8 resin, and the eluent was a 30% (v / v) ethanol solution; the elution time was 1 h, and the eluent flow rate was 3 mL / min. The active substance was dissolved in dichloromethane to obtain an active substance solution. The active substance solution was adsorbed onto a silica gel column. The adsorbed silica gel column was eluted with the eluent used in silica gel column chromatography. The resulting eluent was collected, and the same components were concentrated under reduced pressure (20 mmHg) for 60 min to obtain the substance obtained by silica gel column chromatography. The silica gel column used for silica gel column chromatography was a glass column packed with 200-300 mesh silica gel. The eluent used was petroleum ether and ethyl acetate. The concentration of the active substance solution was 100 mg / mL. The elution time was 3 h. The elution program was as follows: elution was performed sequentially with an eluent in a volume ratio of petroleum ether to ethyl acetate of 5:1 for 60 min, with an eluent in a volume ratio of petroleum ether to ethyl acetate of 3:1 for 60 min, and with an eluent in a volume ratio of petroleum ether to ethyl acetate of 1:1 for 60 min until the target compound was completely eluted. The flow rate of the eluent was 3 mL / min. The substance obtained by silica gel column chromatography was dissolved in methanol to obtain an injection solution. This injection solution was then injected into a semi-preparative liquid chromatograph. The individual fractions eluting from the column were monitored, and the positions of the furanocoumarin compounds were determined based on their retention times. The corresponding fractions were collected using a fractionating distiller or an automatic collection system. The obtained fractions were concentrated under reduced pressure (20 mmHg) for 60 min to remove the solvent, yielding the furanocoumarin compounds. These compounds were then freeze-dried at -50℃ for 20 h, yielding a purity of 97% and a yield of 0.008% (based on the weight of dried Angelica dahurica root powder). The monitoring method used was ultraviolet (UV) detection. The semi-preparative liquid chromatographic separation conditions were as follows: Chromatographic column: C18 column, dimensions: 250 mm × 10 mm, particle size: 10 µm, mobile phase composition (volume ratio): acetonitrile:water = 55:45, flow rate: 2 mL / min, column temperature: 22 °C, detection wavelength: 254 nm, injection volume: 1000 µL. The concentration of the injection solution was 1 mg / mL. The purity of the obtained furanocoumarin compounds was 97%, and the yield was 0.008%.

[0047] Example 2 The difference from Example 1 is that the ultrasonic extraction time is 60 min, while the other conditions are the same as in Example 1.

[0048] Test results: Purity of furanocoumarins: 97.5%; Yield: 0.0095%; Furanocoumarin content in extract: 2.15 mg / g Conclusion: Extending the sonication time to 60 min increased the extraction efficiency by about 18%, but the purity did not change much, indicating that a sonication time of 60 min can extract the target compound more fully.

[0049] Example 3 The difference from Example 1 is that the ultrasonic extraction power was 400W and the time was maintained for 30 minutes, while other conditions were the same as in Example 1.

[0050] Test results: Purity of furanocoumarins: 96.5%; Yield: 0.0092%; Furanocoumarin content in the extract: 2.08 mg / g Conclusion: Increasing the ultrasonic power to 400W improved the extraction efficiency by about 15%, but the purity decreased slightly (possibly due to the co-extraction of some impurities caused by the high power), proving that the 400W power is within an acceptable range.

[0051] Comparative Example 1 The traditional reflux extraction method was employed. Specifically, 100g of Angelica dahurica root powder was added to 1500mL of 95% ethanol, and the mixture was refluxed at 80℃ for 2 hours. This extraction was repeated three times. The extracts were combined and concentrated under reduced pressure to obtain a paste. This paste was then subjected to preliminary separation using macroporous resin column chromatography, silica gel column chromatography, and semi-preparative liquid chromatography. The final product contained furanocoumarin compounds with a purity of 89% and a yield of 0.005%.

[0052] Comparative Example 2 Ultrasonic-assisted extraction was employed, but the ultrasonic parameters were not optimized. The specific procedure was as follows: 100g of Angelica dahurica root powder was added to 1500mL of 95% ethanol, and ultrasonic extraction was performed for 30min at 150W, 15kHz, and 80℃. This extraction was repeated three times. The extracts were combined and concentrated under reduced pressure to obtain a paste. This paste was then subjected to preliminary separation using macroporous resin column chromatography, silica gel column chromatography, and semi-preparative liquid chromatography. The final purity of the furanocoumarin compounds was 92%, with a yield of 0.006%.

[0053] Comparative Example 3 Optimized ultrasonic extraction was employed, but preliminary separation using a macroporous resin column was not performed. Specifically, 100g of Angelica dahurica root powder was added to 1500mL of 95% ethanol, and ultrasonic extraction was performed for 30min at 300W, 20kHz, and 60℃. This extraction was repeated three times. The extracts were combined and concentrated under reduced pressure to obtain a paste, which was then directly separated by silica gel column chromatography and semi-preparative liquid chromatography. The final product contained furanocoumarin compounds with a purity of 88% and a yield of 0.007%.

[0054] The comparative experiments of Example 1 and Comparative Examples 1-3 show that although the traditional reflux extraction method (Comparative Example 1) is simple to operate, its extraction efficiency is low, and prolonged high-temperature heating easily leads to the degradation of heat-sensitive components, resulting in low purity and yield of the final product. When ultrasound-assisted extraction was used without parameter optimization (Comparative Example 2), although purity and yield improved, the extraction efficiency was still unsatisfactory because the ultrasound power and frequency were not at their optimal values. When the preliminary separation step using a macroporous resin column was omitted (Comparative Example 3), although the yield slightly improved, the burden of subsequent purification increased due to the ineffective removal of impurities, resulting in a significant decrease in the purity of the final product. Only by using the complete process flow of this invention (Example 1), i.e., optimizing the ultrasonic extraction parameters and combining preliminary separation with macroporous resin column, silica gel column chromatography, and semi-preparative liquid chromatography, can high purity (97%) and a relatively high yield (0.008%) of furanocoumarin compounds be obtained.

[0055] In summary, by optimizing the extraction process and purification procedure, this invention can efficiently prepare high-purity furanocoumarin compounds, which is significantly superior to traditional methods and other improved methods.

[0056] Performance testing Figure 1 The effects of compounds from Angelica dahurica on the secretion levels of NO, TNF-α, IL-6, and IL-1β in LPS-induced RAW 264.7 macrophages were investigated. The control group (con / control) represented the normal group, and the LPS group (LPS-induced inflammation) represented the lipopolysaccharide group. The compounds were: 2-Methyl-,2-[(8S)-8,9-dihydro-2-oxo-2H-furo[2,3-h]chromen-8-yl]allyl ester-2-(Z)-butenoic acid (1); (-)-(3'S,4'S)-3'-acetoxy-4'-angeloyloxy-3',4'-dihydroseselin (2); praeruptorin D (3); decursin (10); and (+)-trans-khellactone (13). (13), comp.20 is isoimperatorin (20), comp.28 is skimmin (28), and comp.29 is vanillic acid (29).

[0057] from Figure 1It can be found that furanocoumarins in Angelica dahurica can significantly inhibit the secretion of NO, TNF-α, IL-1β and IL-6 in LPS-induced RAW 264.7 macrophages, and have anti-inflammatory effects.

[0058] Figure 2The study investigated the effects of compounds from Angelica dahurica on LPS-induced RAW 264.7 macrophage activity. The control group (con) was untreated, and the LPS group (inflammation induced by LPS) was used. The compounds were: 1. 2-Methyl-,2-[(8S)-8,9-dihydro-2-oxo-2H-furo[2,3-h]chromen-8-yl]allyl ester-2-(Z)-butenoic acid (1); 2. (-)-(3'S,4'S)-3'-acetoxy-4'-angeloyloxy-3',4'-dihydroseselin (2); 3. praeruptorin D (3); 4. praeruptorin A (4); and 5. praeruptorin D (4). B(5), comp.6 is peuformosin(6), comp.7 is (+)-samidin(7), comp.8 is (±)-cis-4'-tigloylkhellactone(8), comp.9 is 7-methoxy-8-β-glucopyranosyl coumarin(9), comp.10 is decursin(10), comp.11 is hyuganin D(11), comp.12 is selinidin(12), comp.13 is (+)-trans-khellactone(13), comp.14 is (-)-cis-khellactone(14), comp.15 is angenomalin(15), comp.16 is imperatorin(16), comp.17 is isobyakangelicin hydrate-3'-ethylether (17), comp.18 is neobyakangelicol (18), comp.19 is phellopterin (19), comp.20 is isoimperatorin (20), comp.21 is marmesin (21), comp.22 is bergapten (22), comp.23 is adicardin (23), comp.24 is umbelliferone (24), comp.25 is scopoletin (25), comp.26 is isoscopoletin (26), comp.27 is fraxidin 8-O-β-D-glucopyranoside (27), comp.28 is skimmin (28), comp.29 is vanillic acid (29), comp.Comp. 30 is 4-(2-hydroxy-1-methoxyethyl)phenol (30), comp. 31 is daucosterol (31), comp. 32 is 3-O-feruloylquinicacid methyl ester (32), comp. 33 is (11S,16S,Z)-Octadeca-9,17-dien-12,14-diyne-1,11,16-triol (33), comp. 34 is oplopantriol A (34), and comp. 35 is n-tetradecanyl oleate (35).

[0059] from Figure 2 It can be seen that furanocoumarin compounds in Angelica dahurica can significantly enhance the activity of LPS-induced RAW 264.7 macrophages within a certain concentration range, indicating that they have the effect of protecting macrophages from inflammatory damage.

[0060] Figure 3 The effects of compounds 2 and 3 on LPS-induced NF-κB signaling pathway in RAW 264.7 cells were investigated. LPS was a lipopolysaccharide group, compound 2 was (-)-(3'S,4'S)-3'-acetoxy-4'-angeloyloxy-3',4'-dihydroseselin, and compound 3 was praeruptorin D. From... Figure 3 It can be seen that compounds 2 and 3 can significantly inhibit the activation of the NF-κB signaling pathway in LPS-induced RAW264.7 cells, indicating that their anti-inflammatory effect is related to the inhibition of the NF-κB signaling pathway.

[0061] Figure 4 The effects of compounds 2 and 3 on LPS-induced MAPK signaling pathway in RAW 264.7 cells were investigated. LPS was a lipopolysaccharide compound, compound 2 was (-)-(3'S,4'S)-3'-acetoxy-4'-angeloyloxy-3',4'-dihydroseselin, and compound 3 was praeruptorin D. From... Figure 4 It can be seen that compounds 2 and 3 can inhibit LPS-induced phosphorylation of the MAPK signaling pathway (including p38, ERK1 / 2 and JNK) in RAW 264.7 cells, further confirming that they exert anti-inflammatory effects by inhibiting the MAPK signaling pathway.

[0062] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A furanocoumarin compound, characterized in that, It has the structure shown in Equation I: Equation I.

2. The method for preparing the furanocoumarin compound according to claim 1, characterized in that, Includes the following steps: Angelica root powder and extraction solvent were mixed and subjected to ultrasonic extraction. The resulting extract was concentrated under reduced pressure to obtain an extract. The extraction solvent was a 70% (v / v) aqueous ethanol solution. The extract was initially separated by a macroporous resin column, and the resulting active substances were then separated by silica gel column chromatography and semi-preparative liquid chromatography to obtain the furanocoumarin compounds. The resin in the macroporous resin column is AB-8 resin.

3. The preparation method according to claim 2, characterized in that, The particle size of the Angelica dahurica root powder is 10-60 mesh.

4. The preparation method according to claim 2, characterized in that, The ultrasonic extraction was performed at a power of 200-400W, a frequency of 20-30Hz, a temperature of 50-70℃, and a time of 30-60min.

5. The preparation method according to claim 2, characterized in that, The pressure for vacuum concentration is 10~30 mmHg, the temperature is 35~45℃, and the time is 40~80 min.

6. The preparation method according to claim 2, characterized in that, The eluent used in the silica gel column chromatography is selected from petroleum ether and ethyl acetate, or petroleum ether and dichloromethane, or dichloromethane and methanol.

7. The preparation method according to claim 2, characterized in that, The conditions for the semi-preparative liquid chromatography separation include: Chromatographic column: C18 column, dimensions: 250mm × 10mm, packing material particle size: 5~10µm. The mobile phase composition by volume ratio is: acetonitrile:water = 55~60:40~45. Flow rate: 1~2 mL / min Column temperature: 20~25℃ Detection wavelength: 254nm Injection volume: 500~1500µL, concentration of injection solution: 0.5~2mg / mL.

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