An adenosine compound in purslane and its extraction and separation method and use

Abstract

The present invention belongs to the technical field of traditional Chinese medicine extraction and separation, and particularly relates to an adenosine compound in purslane, and its extraction, separation method and use. The adenosine compound has a molecular formula of C 12 H 15 N5O5, and is named N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9 H -purin-6-yl)acetamide 5-hydroxypicolinamide. The present invention also provides a method for extracting and separating the above adenosine compound, which sequentially uses water decoction extraction, macroporous resin column chromatography, ODS column chromatography and high performance liquid chromatography for separation, purification and preparation. Its structure is determined to be a new natural product by methods such as mass spectrometry, hydrogen nuclear magnetic resonance spectroscopy, carbon nuclear magnetic resonance spectroscopy and two-dimensional nuclear magnetic resonance spectroscopy. The adenosine compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9 H -purin-6-yl)acetamide 5-hydroxypicolinamide has anti-inflammatory activity, anti-cholinesterase activity and antioxidant activity, and the compound and its salts or derivatives can be used as raw materials for drug development and pharmacological activity research.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine extraction and separation, and in particular relates to an adenosine compound in purslane and an extraction and separation method and application thereof. Background Art

[0002] Purslane comes from the dried aerial parts of Portulaca oleracea L., a plant of the Portulacaceae family. It is one of the wild plants with both medicinal and edible properties designated by the Ministry of Health of my country. It is also known as long-life vegetable and five-element grass. As a traditional Chinese medicine in my country, Purslane has a history of thousands of years of use. It is widely distributed, rich in resources, and has very strong adaptability and tenacious vitality. The 2020 edition of the "Pharmacopoeia of the People's Republic of China" contains the dried aerial parts of Purslane as medicine. It tastes sour and cold, and enters the liver and large intestine meridians. It has the effects of clearing away heat and detoxifying, cooling blood and stopping bleeding, and stopping dysentery. It is used for heat-toxic bloody dysentery, carbuncle, furuncle, eczema, erysipelas, snake and insect bites, blood in the stool, hemorrhoids, and metrorrhagia.

[0003] Purslane contains a variety of chemical components, mainly including: alkaloids, flavonoids, organic acids, terpenes, coumarins, polysaccharides, amino acids and minerals. Among them, there are many types of alkaloid compounds in Purslane, such as uracil, adenine, norepinephrine, dopamine, adenosine, N,N-dicyclohexylurea, purslanamide, allantoin, N-trans-feruloyltyramine, p-hydroxyphenylethylamine, Oleracein AE, Oleraindole AG, Oleraisoindole, Portulaceramide A, etc. The chemical components contained in Purslane are closely related to its diverse pharmacological effects. Modern pharmacological studies have shown that Purslane has antioxidant, neuroprotective, anti-inflammatory, antibacterial, anti-tumor, hypoglycemic, liver-protecting and immune-enhancing effects. There are many kinds of chemical components in Purslane and their pharmacological activities are diverse. The development and separation of compounds in Purslane can provide a basis for in-depth research on Purslane. Summary of the invention

[0004] In view of the above problems, the present invention provides an adenosine compound extracted from Portulaca oleracea. Studies have shown that the adenosine compound of the present invention has anti-inflammatory activity, anticholinesterase activity and antioxidant activity. At the same time, a simple, rapid, environmentally friendly and high-purity extraction and separation method for the compound of the present invention is provided.

[0005] To achieve the above object, the present invention provides an adenosine compound having a molecular formula of C 12 H 15N5O5, named N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide according to its structure, has the following chemical formula:

[0006]

[0007] The present invention also provides a method for extracting and separating adenosine compounds, which specifically comprises the following steps:

[0008] Step 1: Take dried purslane, decoct in water to extract, concentrate the extract, cool to room temperature, and set aside the liquid medicine.

[0009] Step 2: The concentrated solution in step 1 is subjected to a macroporous resin, and eluted with water and ethanol of different concentrations, and the 30% ethanol portion is recovered under reduced pressure to obtain an extract, and a concentrate is obtained for use.

[0010] Step 3: Separate the concentrate in step 2 through an ODS column (Octadecylsilyl, octadecylsilane bonded silica gel filler) using a methanol-water gradient elution, detect by thin layer chromatography, develop color, combine the 30% methanol elution portion and evaporate to dryness to obtain a concentrate for use.

[0011] Step 4: The concentrate obtained in step 3 is separated by ODS column chromatography again, using methanol-water gradient elution, detected by thin layer chromatography, color development, and the 50% methanol elution portion is combined and evaporated to dryness to obtain a concentrate for use.

[0012] Step 5: The concentrate obtained in step 4 is separated and prepared by HPLC (high performance liquid chromatography), and isocratic elution is performed using methanol-0.1% formic acid (volume percentage) as the mobile phase to finally obtain the compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide described in the present invention.

[0013] Furthermore, in step 1, the water decocting and extraction are performed twice, each time for 2 hours, and the amount of water used is 8 to 16 times the amount of the medicinal material.

[0014] Furthermore, in step 2, the macroporous resin is AB-8 macroporous resin, and the volume ratio of ethanol to water is 30:70, 50:50, 70:30 and 100:0 for gradient elution.

[0015] Furthermore, in step 3, the volume ratio of methanol to water is 10:0, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40 to 70 μm.

[0016] Furthermore, in step 4, the volume ratio of methanol to water is 10:0, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40 to 70 μm.

[0017] Furthermore, the ODS pretreatment process in step 3 and step 4 is: soaking in methanol for 24 hours, after loading on the column, washing with methanol until there is no turbidity when dripped into water, and then balancing with the initial mobile phase.

[0018] Furthermore, in step 5, the volume ratio of methanol to 0.1% formic acid is 10:90, and the retention time of the compound is 8.141 min.

[0019] The present invention also provides a use of the adenosine compounds separated from the purslane medicinal material as described above in the preparation of anti-inflammatory drugs, anticholinesterase drugs and antioxidant drugs.

[0020] Compared with the prior art, the invention has the beneficial effects.

[0021] The separation and pharmacological activity research of the adenosine compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide in purslane described in the present invention has not been reported in plants; the present invention provides the adenosine compound derived from purslane and a method for extracting and separating the compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention, which successively adopts water decoction extraction, macroporous resin column chromatography, ODS column chromatography and high performance liquid chromatography for separation, purification and preparation, and successfully extracts and separates an adenosine compound. The method The method has only five steps, the operation method is simple and fast, water is used for extraction, the process method is environmentally friendly, and the purity of the compound separated by the method is high, greater than 90%; in addition, studies have shown that the compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide has anti-inflammatory activity, anti-cholinesterase activity and antioxidant activity; therefore, the new natural compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide and its salts and derivatives separated by the present invention can be used as raw materials for drug development and pharmacological activity research. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention 1 H-NMR spectrum.

[0023] Figure 2 The N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention 13 C-NMR spectrum. DETAILED DESCRIPTION

[0024] The following examples will help to understand the present invention, but these examples are only for illustrating the present invention, and the present invention is not limited to these contents. The operating methods in the examples are all conventional operating methods in the art.

[0025] Example 1.

[0026] The present invention provides an adenosine compound, the molecular formula of which is C 12 H 15 N5O5, named N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide, has the chemical structure:

[0027]

[0028] Table 1 Adenosine compounds

[0029] NMR data of N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide. The solvent used for NMR is DMSO-d6.

[0030] Table 1 NMR data of N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide

[0031]

[0032]

[0033] N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)aceta mide: brown powder, easily soluble in water, slightly soluble in methanol. After being spotted on a silica gel thin layer plate, the spots appear orange-yellow when sprayed with dilute potassium bismuth iodide test solution, indicating that the compound is an alkaloid component. 1 H-NMR signals showed that δ H 8.34(1H,s,H-2),δ H 8.23 (1H, s, H-7), δ H 2.95(3H,s,H-12),δ H 5.88(1H,d,J=6.24Hz,H-2'), δ H 4.60(1H,t,J=5.46,11.04Hz,H-3'), δ H 4.13 (1H, m, H-4'), δ H 3.96(1H,m,H-5'),δ H 3.67(1H,dd,J=3.54,12.12Hz,H-6'),δ H 3.54(1H,dd,J=3.54,12.12Hz,H-6'); 13 C-NMR signals showed that C-2(δ C 155.09), C-4(δ C 152.49), C-5(δ C 119.95), C-7(δ C 139.73), C-9(δ C 148.06), C-11(δ C 163.56), C-12(δ C 27.04), C-2(δ C 155.09), C-2'(δ C 87.95), C-3'(δ C 73.51), C-4'(δ C 70.72), C-5'(δ C 85.97), C-6'(δ C 61.73). The above signals were compared with the literature, except for δ H 2.95(3H,s,H-12),C-11(δ C 163.56) and C-12 (δ C27.04), the remaining signals are consistent with the adenosine structure displacement reported in Chemistry of natural compounds. 2007, 43(2): 181. Based on the adenosine structure core, it is speculated that the amino position is substituted with an acetyl group. Based on the above information, it can be determined that this new natural product has the above structure.

[0034] The present invention also provides a method for extracting and separating N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide, which comprises the following specific steps:

[0035] Step 1: Weigh 250 kg of dried purslane medicinal materials, use water decoction to extract, the amount used is 10 times the medicinal material, extract twice, each time for 2 hours, combine the extracts, heat and concentrate, cool to room temperature, and obtain the medicinal solution for use.

[0036] Step 2: evaporate the drug solution obtained in step 1 to dryness, separate it through AB-8 macroporous resin column chromatography, use ethanol-water (30:70, 50:50, 70:30 and 100:0, v:v) gradient elution, collect the 30% ethanol portion, recover it to the extract under reduced pressure, and obtain the concentrate for use.

[0037] Step 3: Separate the concentrate in step 2 through a pretreated ODS column with a filler particle size of 40 to 70 μm, and use methanol-water (10:0, 30:70, 50:50, 70:30 and 100:0, v:v) gradient elution, detect by thin layer chromatography, develop color, combine the 30% methanol elution fractions, concentrate to dryness under reduced pressure, and set aside.

[0038] Step 4: The obtained product in step 3 is separated by pre-treated ODS column chromatography, the filler particle size is 40-70 μm, and gradient elution is performed with methanol-water (10:0, 30:70, 50:50, 70:30 and 100:0, v / v), and detection is performed by thin layer chromatography, color development, and the 50% methanol elution part is combined, concentrated to dryness under reduced pressure, and used for standby. The ODS pretreatment process is to soak in methanol for 24 hours, after loading the column, wash with methanol until there is no turbidity when dripped into water, and then balance with the initial mobile phase.

[0039] Step 5: The fraction obtained in step 4 was separated and prepared by HPLC, with methanol and 0.1% formic acid in a volume ratio of 10:90 as the mobile phase and detection wavelengths of 210 and 254 nm to obtain the adenosine compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention; the purity was determined by normalization method to be 96%.

[0040] Example 2.

[0041] Anti-inflammatory effect of N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)ac etamide

[0042] 1 Main Materials

[0043] 1.1 Drugs and reagents: The new natural product used in the experiment was prepared by the above method with a purity of 96%. DMEM high-glucose medium, fetal bovine serum (Hyclone, USA); penicillin and streptomycin (Hangzhou Sijiqing Company); LPS (Sigma, USA); IL-1β ELISA kit (Cayman, USA); cell lysate.

[0044] 1.2 Cell line: RAW264.7 macrophages (ATCC cell bank, USA).

[0045] 1.3 Grouping: Divided into control group, LPS group and experimental group.

[0046] 2 Experimental methods.

[0047] 2.1 Cell culture: Add 10% fetal bovine serum and 1% antibiotics (100 U / mL penicillin and 100 μg / mL streptomycin) to DMEM high-glucose medium and store at 4°C.

[0048] 2.2 CCK-8 assay for cell viability: The three groups of RAW264.7 macrophages in logarithmic growth phase were inoculated in 96-well culture plates at a cell density of 1×10 4 / mL, 100μL per well, 37℃, 5% CO2, after overnight culture, different concentrations of the compound of the present invention (5μM~50μM) were added to the experimental group, and after 1h of incubation, 1μg / mL of LPS was added to the LPS group and the experimental group, and a zero adjustment group (culture solution containing DMSO solvent) was set up, and 3 replicates were set up in each group to investigate the effect of the addition of drugs on the cells. After the cells in the above groups were cultured for 24h, 10μL of CCK-8 was added to the cells in each well, and after further incubation for 4h at 37℃, 5% CO2, the absorbance of each well was measured at a wavelength of 450nm by an enzyme marker.

[0049] 2.3ELISA method for determination of inflammatory factor IL-1β: RAW264.7 macrophages in the logarithmic growth phase were inoculated in 24-well culture plates at a cell density of 1×10 5 / mL, 1mL per well, cultured overnight at 37°C and 5% CO2, the experimental group added the compound of the present invention N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide (1μM~20μM) and cultured for 1h, then LPS (final concentration of 1μg / mL) was added to each well and incubated for 24h, and each group was treated with 3 wells. The content of IL-1β was determined by ELISA.

[0050] 3 Experimental results.

[0051] The experimental results show that the compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention has no effect on the proliferation of LPS-induced macrophages RAW264.7; it can effectively inhibit the secretion of excessive inflammatory cytokine IL-1β produced by LPS-induced macrophages RAW264.7 in a concentration-dependent manner.

[0052] The results of the relative cell survival rate experiment are shown in Table 3.

[0053] Table 2 Effect of compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide on the relative survival rate of RAW264.7 macrophages.

[0054]

[0055] The results of ELISA determination of inflammatory factor IL-1β are shown in Table 3.

[0056] Table 3 Compounds

[0057] Effect of N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide on the IL-1β content secreted by RAW264.7 cells induced by LPS (mean ± SD, n = 3).

[0058]

[0059]

[0060] Note: * P<0.05 compared with the control group.# P<0.05 compared with LPS group.

[0061] Example 3.

[0062] Anticholinesterase Effect of N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide

[0063] 1Main materials.

[0064] 1.1 Drugs and reagents: The new natural products used in the experiment were prepared by the above method with a purity of 96%. Physostigmine (Shanghai Hanxiang Biotechnology Co., Ltd.), acetylcholine iodide (ATCI) and acetylcholinesterase (AChE) (Dalian Meilun Biotechnology Co., Ltd.), dithiodinitroformic acid (DTNB) (Shanghai Jinshui Biotechnology Co., Ltd.), disodium hydrogen phosphate and sodium dihydrogen phosphate (Shanghai Sinopharm Reagent Co., Ltd.).

[0065] 1.2 Experimental instruments and equipment: HBS-1096A96-well microplate reader (Nanjing Detie Experimental Equipment Co., Ltd.), 1 / 100,000 balance (METTLER, Switzerland), HH-4 digital display constant temperature water bath (Jiangsu Jintan Ronghua Instrument Manufacturing Co., Ltd.).

[0066] 2 Experimental methods.

[0067] This experiment determines the anticholinesterase activity of each compound according to the modified Ellman method. The compound of the present invention and physostigmine were accurately weighed and prepared into five series of sample solutions with concentrations of 31.25μM, 62.5μM, 125μM, 250μM and 500μM with methanol. The specific operation is as follows: 140μL of phosphate buffered saline (0.1M, pH=8.0, containing 0.1mol / L disodium hydrogen phosphate and sodium dihydrogen phosphate), 20μL of sample solution, 15μL of AChE (0.2U / mL) were added to a 96-well ELISA plate, and after incubation at 37°C for 10 minutes, 10μL of ATCI (4mmol / L) and 10μL of DTNB (15mmol / L) were added, and after incubation at 37°C for 20 minutes, the 96-well plate was placed in an ELISA instrument, and the absorbance value of each group of samples was measured at a wavelength of 405nm. Among them, methanol was used instead of the sample solution as the blank group, and physostigmine was used instead of the sample solution as the positive control group. The cholinesterase inhibition rate of each compound was calculated according to the following formula (A represents absorbance):

[0068] Inhibition rate (%) = (A 空白 -A 样品 ) / A 空白 ×100%

[0069] 3 Experimental results.

[0070] The experimental results show that the new natural product N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention exhibits certain anticholinesterase activity, and the inhibitory effect on cholinesterase increases with the increase of the compound concentration, showing a dose-dependent trend.

[0071] The anticholinesterase activity of the new natural product N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention is shown in Table 4.

[0072] Table 4 Anticholinesterase activity of compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide.

[0073]

[0074] Example 4.

[0075] Antioxidant effect of N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)ac etamide.

[0076] 1 Main Materials

[0077] 1.1 Drugs and reagents: The new natural products used in the experiment were prepared by the above method with a purity of 96%. 1,1-Diphenyl-2-picrylhydrazyl free radical (DPPH) (Sigma, USA), butylated hydroxyanisole (BHA) (Shanghai Xiangrui Biological Co., Ltd.), methanol (chromatographic grade, Tianjin Kaixin Chemical Industry Co., Ltd.)

[0078] 1.2 Experimental instruments and equipment: Hitachi UV-3010 ultraviolet-visible spectrophotometer (Hitachi, Japan), 1 / 100,000 balance (METTLER, Switzerland).

[0079] 2 Experimental methods.

[0080] This experiment uses the DPPH free radical scavenging method to determine the antioxidant activity of each compound. Accurately weigh the compound of the present invention and BHA, and use methanol to prepare five series of sample solutions with concentrations of 12.5μM, 25μM, 50μM, 100μM and 200μM. In addition, the DPPH solution is prepared as needed, and an appropriate amount of DPPH is accurately weighed and used to prepare a solution with a concentration of 80μM with methanol. The whole process is protected from light. The specific operation is as follows: 1mL of the sample solution is fully mixed with 1mL of the DPPH solution, and placed at room temperature and in a light-proof environment for 10min. After setting the detection wavelength of the ultraviolet spectrophotometer to 517nm, the absorbance value of the sample is measured. Among them, methanol is used instead of the sample solution as the blank group, BHA is used instead of the sample solution as the positive control group, and a mixed solution of 1mL of methanol and 1mL of the sample solution is used as the control group. The DPPH scavenging rate of each compound is calculated according to the following formula (A represents absorbance):

[0081] DPPH clearance rate (%) = (1-(A 样品 -A 对照 ) / A 空白 )×100%

[0082] 3 Experimental results.

[0083] The experimental results show that the new natural product N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention exhibits certain antioxidant activity, and the antioxidant effect increases with the increase of the compound concentration, showing a dose-dependent trend.

[0084] The antioxidant activity of the new natural product N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide of the present invention is shown in Table 5.

[0085] Table 5 Antioxidant activity of compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide.

[0086]

[0087]

[0088] In summary, the present invention provides a compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide and an extraction and separation method thereof, which uses water decoction extraction, macroporous resin column chromatography, ODS column chromatography and high performance liquid chromatography for separation, purification and preparation, and successfully extracts and separates an adenosine compound. The operation method is simple, rapid and environmentally friendly, and the compound separated by the method has a high purity. Since the obtained compound is extracted from the commonly used traditional Chinese medicine Portulaca oleracea, it has anti-inflammatory activity, anticholinesterase activity and antioxidant activity, so the new natural product and its salt and derivatives of the present invention can provide ideas for drug development and have broad development prospects.

Claims

1. A method for extracting and separating adenosine compounds from purslane medicinal materials, characterized in that: The specific steps include: Step 1: Take dried purslane medicinal material, decoct it in water, concentrate the extract, cool it to room temperature, and obtain the medicinal solution for later use; Step 2: The concentrated solution in step 1 is subjected to a macroporous resin, and eluted with water and ethanol of different concentrations, and the 30% ethanol portion is recovered under reduced pressure to obtain an extract, and a concentrate is obtained for standby use; Step 3: Separate the concentrate in step 2 through an ODS column, use a methanol-water gradient elution, detect by thin layer chromatography, develop color, combine the 30% methanol elution portion and evaporate to dryness to obtain a concentrate for use; Step 4: The concentrate obtained in step 3 is separated by ODS column chromatography, eluted by methanol-water gradient, detected by thin layer chromatography, and color developed. The 50% methanol elution portion is combined and evaporated to dryness to obtain a concentrate for use; Step 5: The concentrate obtained in step 4 is separated and prepared by HPLC, and isocratic elution is performed using volume percentage methanol-0.1% formic acid as the mobile phase to finally obtain the adenosine compound N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide; The molecular formula of the adenosine compound is: 12 H 15 N5O5, named according to the structural formula, is N-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acetamide, and its chemical structure is: 。 2. The extraction and separation method according to claim 1, characterized in that: In the step 1, the extraction is performed by decoction twice, each time for 2 hours, and the amount of water used is 8 to 16 times the amount of the medicinal material.

3. The extraction and separation method according to claim 1, characterized in that: In the step 2, the macroporous resin is AB-8 macroporous resin, and the volume ratio of ethanol to water is 30:70, 50:50, 70:30 and 100:0 for gradient elution.

4. The extraction and separation method according to claim 1, characterized in that: In the step 3, the volume ratio of methanol to water is 10:0, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40-70 μm.

5. The extraction and separation method according to claim 1, characterized in that: In the step 4, the volume ratio of methanol to water is 10:0, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40-70 μm.

6. The extraction and separation method according to claim 1, characterized in that: In the step 5, the volume ratio of methanol to 0.1% formic acid is 10:90, and the retention time of the compound is 8.141 min.

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