Humulus lupulus xanthohumol derivative as well as preparation method and application thereof

Abstract

The invention provides a xanthohumol derivative in humulus lupulus as well as a preparation method and application thereof.The xanthohumol derivative is obtained by detecting and analyzing female flowers and fruits of humulus lupulus by thin-layer chromatography and analytical high-performance liquid chromatography, extracting with an organic solvent, and concentrating to obtain the xanthohumol derivative. And then separating through a solvent extraction method, macroporous adsorption resin column chromatography, normal-phase silica gel column chromatography, medium-pressure preparative chromatography, Sephadex LH-20 gel column chromatography and semi-preparative high performance liquid chromatography to obtain 22 new xanthohumol derivatives. 23 monomers are determined to be new xanthohumol derivatives through methods of high-resolution mass spectrum, nuclear magnetic resonance spectrum, circular dichroism spectrum, quantum chemistry calculation and the like, and structural identification is carried out on the new xanthohumol derivatives. Besides, the minimum concentration (MIC90) of Mycobacterium tuberculosis (Mtb) H37Ra is inhibited to evaluate the inhibition effect of the xanthohumol derivative on the Mtb H37Ra strain, and the xanthohumol derivative can be used for preparing antituberculosis drugs.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to xanthohumol derivatives in hops and preparation methods and uses thereof. Background Art

[0002] Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb), a chronic infectious disease that has long plagued mankind, is still one of the major public health problems facing the world. It is a preventable and curable disease. However, according to data released by the World Health Organization, in 2022, tuberculosis will still be the world's second largest single infectious cause of death, second only to the coronavirus, and the number of deaths caused is almost twice that of HIV. Tuberculosis is caused by Mycobacterium tuberculosis, and the emergence of multidrug resistance, extensive drug resistance and complete drug resistance has made most drugs on the market ineffective, which has brought new challenges to the treatment of tuberculosis. Therefore, it is necessary to develop anti-tuberculosis drugs with new mechanisms of action to solve the serious problem of drug resistance.

[0003] Hops (Humulus lupulus L.), also known as Hop (English common name Hop), Humulus lupulus, and Hops, are perennial climbing herbs belonging to the genus Humulus of the subfamily Cannabinoides. They are one of the indispensable raw materials for beer brewing. They give beer a unique bitterness and aroma, and have the ability to preserve and clarify wort. They are known as the soul of beer. In addition, hops are also an important traditional medicinal material. Originated from Europe, they grow wild in Xinjiang, my country, and are cultivated in Northeast China, North China, Shandong and other places. Xinjiang has sufficient sunlight and a large temperature difference between day and night, and is the main distribution area of ​​wild hops in my country. Its female cone spike inflorescence is not only an additive for brewing beer, but also widely used as a medicine around the world. Hops have the effects of strengthening the stomach, digesting food, diuresis, calming the nerves, relieving cough and reducing phlegm. They can be used to treat loss of appetite, abdominal distension, tuberculosis, pleurisy, insomnia, hysteria, edema, cystitis, etc. As a national medicine (traditional Chinese medicine), it is included in many classics and is a Xinjiang specialty plant resource that can be used as both medicine and food. Among them, the first volume of the 1977 edition of the Chinese Pharmacopoeia records Sanhesu ointment and Sanhesu capsules (the main raw material is hop extract), which have antibacterial and anti-inflammatory effects and are used to treat pulmonary tuberculosis, tuberculous pleurisy, and leprosy.

[0004] Xanthohumol is a simple isopentenyl-containing chalcone (accounting for about 0.1% to 1% of the dry weight of hops). In fact, hops are the only source of natural isopentenyl-containing flavonoids found so far, such as xanthohumol and isoxanthohumol, which exist in plants in the form of aglycones. Hops are one of the main raw materials for beer brewing, which can give beer a refreshing bitter taste and aroma. Beer has become the main dietary source of xanthohumol and other isopentenyl-containing flavonoids. The hard resin component of hops, xanthohumol and its derivative, isoxanthohumol, are also beneficial to the stability of beer foam. It is well known that flavonoid compounds have a wide range of physiological and pharmacological activities, including choleretic, anti-ulcer, hepatoprotective, antispasmodic, diuretic, antiviral, etc. Isopentenyl-containing flavonoids also have anti-proliferative and anti-cancer effects.

[0005] However, up to now, there has been no report on the anti-mycobacterium tuberculosis effect of xanthohumol derivatives in hops.

[0006] Among them, the disclosed invention patent: A xanthohumol derivative and its preparation method and use (patent number: ZL2020 10957790.8) is the preliminary work of this subject. This type of derivative uses 1″,2″-dihydroxanthohumol C, xanthohumol C, 1″,2″-dihydroxanthohumol K as raw materials, and obtains a series of compounds through chemical synthesis modification and conducts preliminary anti-inflammatory activity tests. Based on the previous work, the present invention uses phytochemical methods to track and separate chemical components similar to xanthohumol in hops according to ultraviolet absorption characteristics to obtain 22 monomer compounds. Summary of the invention

[0007] The present invention aims to provide xanthohumol and its derivatives in hops, and a preparation method and use thereof. The xanthohumol derivatives are detected and analyzed from the female flowers and fruits of hops by thin layer chromatography and analytical high performance liquid chromatography, extracted with an organic solvent, and then separated by solvent extraction, macroporous adsorption resin column chromatography, normal phase silica gel column chromatography, medium pressure preparative chromatography, Sephadex LH-20 gel column chromatography, and semi-preparative high performance liquid chromatography to obtain 22 new xanthohumol derivatives. The 23 monomers are determined to be new xanthohumol derivatives by high resolution mass spectrometry, nuclear magnetic resonance spectroscopy, circular dichroism spectroscopy and quantum chemical calculation, and their structures are identified. In addition, the minimum concentration (MIC) for inhibiting Mycobacterium tuberculosis (Mtb) H37Ra is used to determine the new xanthohumol derivatives. 90 ) to evaluate the inhibitory effect of xanthohumol derivatives on Mtb H37Ra strain, which can be used to prepare anti-tuberculosis drugs.

[0008] The xanthohumol derivative compound in hops of the present invention has the structural formula:

[0009]

[0010] in:

[0011] The compound of formula (1) is named: 2″R-2′,3′-dihydropyran-4′,4-dihydroxy-6-methylchalcone;

[0012] The compound of formula (2) is named: 2″S-2′,3′-dihydropyran-4′,4-dihydroxy-6-methylchalcone;

[0013] The compound of formula (3) is named: 2″S-3′,4′-dihydropyran-2′,4-dihydroxy-6-methylchalcone;

[0014] The compound of formula (4) is named: 2″R-3′,4′-dihydropyran-2′,4-dihydroxy-6-methylchalcone;

[0015] The compound of formula (5) is named: 2″S-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone;

[0016] The compound of formula (6) is named: 2″R-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone;

[0017] The compound of formula (7) is named: 2″S-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone;

[0018] The compound of formula (8) is named: 2″R-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone;

[0019] The compound of formula (9) is named: 2S, 2″R-7, 8-dihydropyran-4′-hydroxy-5-methyldihydroflavone;

[0020] The compound of formula (10) is named: 2S, 2″S-7, 8-dihydropyran-4′-hydroxy-5-methyldihydroflavone;

[0021] The compound of formula (11) is named: 2S, 2″R-7, 8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone;

[0022] The compound of formula (12) is named: 2S,2″S-7,8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone;

[0023] The compound of formula (13) is named: 2′S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone;

[0024] The compound of formula (14) is named: 2′R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone;

[0025] The compound of formula (15) is named: 2′S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone;

[0026] The compound of formula (16) is named: 2′R-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone;

[0027] The compound of formula (17) is named: 2″R-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone;

[0028] The compound of formula (18) is named: 2″S-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone;

[0029] The compound of formula (19) is named: 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone;

[0030] The compound of formula (20) is named: 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone;

[0031] The compound of formula (21) is named: 6,7-pyran-2-isopropyl-5-hydroxychromone;

[0032] The compound of formula (22) is named: 2'S-8-geranyl-5,7,4'-trihydroxydihydroflavone.

[0033] The method for preparing the xanthohumol derivatives in hops is carried out according to the following steps:

[0034] a. Take female hops as raw materials, dry and crush them, and then use 5-10 times the volume of 50-99% ethanol aqueous solution, anhydrous ethanol, 50-99% methanol aqueous solution or anhydrous methanol to perform percolation, cold soaking or heating reflux extraction at room temperature, and concentrate to obtain a crude extract of hops;

[0035] b. Disperse the crude extract obtained in step a with water, add petroleum ether or n-hexane, chloroform, and n-butanol in sequence, extract for 3-5 times, and concentrate the chloroform extract to obtain a chloroform extract concentrate;

[0036] c. Separating the chloroform extract obtained in step b by macroporous adsorption resin column chromatography, normal phase silica gel column chromatography, medium pressure preparative chromatography, Sephadex LH-20 gel column chromatography and semi-preparative high performance liquid chromatography to obtain compound 1-22;

[0037] The macroporous adsorption resin column chromatography method used is normal pressure or pressurized column chromatography, the filler used is macroporous adsorption resin, the eluent used is 10-90% methanol aqueous solution or ethanol aqueous solution, and isocratic elution or gradient elution is adopted; the normal phase silica gel column chromatography method used is normal pressure column chromatography, the eluent is n-hexane-ethyl acetate mixed solvent, and gradient elution is adopted; the medium pressure preparation chromatography method uses reverse phase C-18 filler, the eluent is methanol aqueous solution or acetonitrile aqueous solution with a volume concentration of 30-100%, and isocratic elution or gradient elution is adopted; the Sephadex LH-20 gel column chromatography method used is normal pressure column chromatography, the eluent is a mixture of methanol, chloroform and water solvents, and isocratic elution is adopted; the preparation high performance liquid chromatography method used is pressurized column chromatography, the eluent is methanol aqueous solution with a volume concentration of 40-99% or acetonitrile aqueous solution with a volume concentration of 30-99%, and isocratic elution or gradient elution is adopted.

[0038] The invention discloses a use of the xanthohumol derivatives in hops in preparing anti-tuberculosis drugs.

[0039] The xanthohumol derivative in hops of the present invention is obtained by separation and purification from plants, and can also be synthesized by chemical modification method.

[0040] The structure of the xanthohumol derivative compound in hops described in the present invention is determined by high-resolution mass spectrometry, one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy, circular dichroism spectroscopy and quantum chemical calculation, and the structural identification process is as follows:

[0041] Compound formula (1) is 2″R-2′,3′-dihydropyran-4′,4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 332nm; ECD (MeOH) 210 (Δε+22.3), 297 (Δε+6.09), 337 (Δε-3.18); the quasi-molecular ion peak [M+H] + m / z 371.1488 (calculated value 371.1416) confirmed its molecular formula to be C 21 H 23 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and it was named as 2″R-2′,3′-dihydropyran-4′,4-dihydroxy-6-methylchalcone. 1 H and 13 C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0042] Compound formula (2) is 2″S-2′,3′-dihydropyran-4′,4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 332nm; ECD (MeOH) 210 (Δε-22.3), 297 (Δε-6.09), 337 (Δε+3.18); the quasi-molecular ion peak [M+H] in its high-resolution mass spectrum + m / z 371.1488 (calculated value 371.1416) confirmed its molecular formula to be C 21 H 23 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and it was named as 2″S-2′,3′-dihydropyran-4′,4-dihydroxy-6-methylchalcone. 1 H and 13 C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0043] Compound formula (3) is 2″S-3′,4′-dihydropyran-2′,4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 372nm; ECD (MeOH) 219 (Δε+9.45), 253 (Δε-6.84), 310 (Δε-9.01), 378 (Δε+3.14); through its high-resolution mass spectrum, the quasi-molecular ion peak [MH] - m / z 369.1340 (calculated value 369.1416) confirmed its molecular formula to be C 21 H 21 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and named as 2″S-3′,4′-dihydropyran-2′,4-dihydroxy-6-methylchalcone. 1 H and 13 C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0044] Compound formula (4) is 2″R-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 372nm; ECD (MeOH) 219 (Δε-9.45), 253 (Δε+6.84), 310 (Δε+9.01), 378 (Δε-3.14); through its high-resolution mass spectrum, the quasi-molecular ion peak [MH] -m / z 369.1340 (calculated value 369.1416) confirmed its molecular formula to be C 21 H 21 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and it was named as 2″R-3′,4′-dihydropyran-2′,4-dihydroxy-6-methylchalcone. 1 H and 13 C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0045] Compound formula (5) is 2″S-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 376nm; ECD (MeOH) 217 ​​(Δε+18.26), 313 (Δε+4.53), 368 (Δε-0.14); through its high-resolution mass spectrum, the quasi-molecular ion peak [MH] - m / z 387.1448 (calculated value 387.1552) confirmed its molecular formula to be C 21 H 23 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and named as 2″S-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone. 1 H and 13 C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0046] Compound formula (6) is 2″R-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 376nm; ECD (MeOH) 218 ​​(Δε-31.19), 286 (Δε-1.42), 366 (Δε+5.38); through its high-resolution mass spectrum quasi-molecular ion peak [MH] - m / z 387.1448 (calculated value 387.1552) confirmed its molecular formula to be C 21 H 23 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and named as 2″R-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone. 1 H and 13C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0047] Compound formula (7) is 2″S-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 372nm; ECD (MeOH) 207 (Δε+8.05), 270 (Δε+3.24), 308 (Δε+0.24), 362 (Δε-1.89); by the quasi-molecular ion peak [MH] in its high-resolution mass spectrum - m / z 369.1314 (calculated value 369.1416) confirmed its molecular formula to be C 21 H 21 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and named as 2″S-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone. 1 H and 13 C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0048] Compound formula (8) is 2″R-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV (ACN) 372nm; ECD (MeOH) 207 (Δε-8.43), 270 (Δε+3.26), 311 (Δε-1.03), 363 (Δε+1.18); through its high-resolution mass spectrum, the quasi-molecular ion peak [MH] - m / z 369.1314 (calculated value 369.1416) confirmed its molecular formula to be C 21 H 21 O6; According to 1 H, 13 The structure of the product was confirmed by C NMR and 2D NMR data and it was named as 2″R-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone. 1 H and 13 C NMR assignments are shown in Table 1 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0049] Compound formula (9) is 2S, 2″R-7,8-dihydropyran-4′-hydroxy-5-methyldihydroflavone; yellow amorphous powder, UV (ACN) 217, 290nm; ECD (MeOH) 217 ​​(Δε+17.20), 236 (Δε+6.91), 289 (Δε-27.2), 334 (Δε+9.37); the quasi-molecular ion peak [MH] in its high-resolution mass spectrum - m / z 369.1331 (calculated value 369.1416) confirmed its molecular formula to be C 21 H 22 O6; According to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2S,2″R-7,8-dihydropyran-4′-hydroxy-5-methyldihydroflavone. 1 H and 13 C NMR assignments are shown in Table 2 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0050] Compound formula (10) is 2S, 2″S-7, 8-dihydropyran-4′-hydroxy-5-methyldihydroflavone; yellow amorphous powder, UV (ACN) 217, 290nm; ECD (MeOH) 217 ​​(Δε-22.60), 236 (Δε-11.10), 289 (Δε+30.17), 335 (Δε-11.26); through its high-resolution mass spectrum, the quasi-molecular ion peak [MH] - m / z 369.1331 (calculated value 369.1416) confirmed its molecular formula to be C 21 H 22 O6; According to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2S,2″S-7,8-dihydropyran-4′-hydroxy-5-methyldihydroflavone. 1 H and 13 C NMR assignments are shown in Table 2 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0051] Compound formula (11) is 2S, 2″R-7,8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone, yellow amorphous powder, UV (ACN) 218, 290nm; ECD (MeOH) 214 (Δε+1.83), 239 (Δε+1.14), 289 (Δε-4.65), 326 (Δε+1.37); through its high-resolution mass spectrum, the quasi-molecular ion peak [MH] - m / z 371.1488 (calculated value 371.1416) confirmed its molecular formula to be C 21 H 23 O6; According to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2S,2″R-7,8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone. 1 H and 13 C NMR assignments are shown in Table 2 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0052] Compound formula (12) is 2S, 2″S-7, 8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone; yellow amorphous powder, UV (ACN) 218, 290nm; ECD (MeOH) 215 (Δε-6.77), 236 (Δε-2.72), 291 (Δε+4.71), 327 (Δε-1.11); through its high-resolution mass spectrum, the quasi-molecular ion peak [MH] - m / z 371.1488 (calculated value 371.1416) confirmed its molecular formula to be C 21 H 23 O6; According to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2S,2″S-7,8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone. 1 H and 13 C NMR assignments are shown in Table 2 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0053] Compound formula (13) is 2′S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; yellow amorphous powder, UV (ACN) 210, 251, 299 nm; ECD (MeOH) 212 (Δε+4.97), 257 (Δε-3.15), 295 (Δε+0.45); the quasi-molecular ion peak [M+H] +m / z 305.1382 (calculated value 305.1311) confirmed its molecular formula to be C 15 H 21 O5; according to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2′S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone. 1 H and 13 C NMR assignments are shown in Table 3 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0054] Compound formula (14) is 2′R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone, yellow amorphous powder, UV (ACN) 210, 251, 299 nm; ECD (MeOH) 212 (Δε-2.92), 256 (Δ+3.12), 285 (Δε-1.44); through the quasi-molecular ion peak [M+H] in its high-resolution mass spectrum + m / z 305.1382 (calculated value 305.1311) confirmed its molecular formula to be C 15 H 21 O5; according to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2′R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone. 1 H and 13 C NMR assignments are shown in Table 3 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0055] Compound formula (15) is 2′S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone, yellow amorphous powder, UV (ACN) 210, 231, 251, 299 nm; ECD (MeOH) 210 (Δε+5.52), 230 (Δε+3.17), 259 (Δε-3.06); through the quasi-molecular ion peak [M+H] in its high-resolution mass spectrum + m / z 319.1541 (calculated value 319.1467) confirmed its molecular formula to be C 18 H 23 O5; according to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2′S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone. 1 H and 13C NMR assignments are shown in Table 2 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0056] Compound (16) is 2′R-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone, yellow amorphous powder, UV (ACN) 210, 231, 251, 299 nm; ECD (MeOH) 209 (Δε+5.52), 230 (Δ+3.17), 259 (Δε+3.06); the quasi-molecular ion peak [M+H] in its high-resolution mass spectrum is + m / z 319.1541 (calculated value 319.1467) confirmed its molecular formula to be C 18 H 23 O5;;According to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2′R-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone. 1 H and 13 C NMR assignments are shown in Table 3 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0057] Compound formula (17) is 2″R-7,8-dihydropyran-4′-hydroxy-5-methyl isoflavone; yellow amorphous powder, UV (ACN) 259nm; ECD (MeOH) 218 ​​(Δε+4.58), 250 (Δε+1.35); through its high-resolution mass spectrum, the quasi-molecular ion peak [M+H] + m / z 369.1325 (calculated value 369.1260) confirmed its molecular formula to be C 21 H 21 O6; According to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2″R-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone. 1 H and 13 C NMR assignments are shown in Table 2 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0058] Compound formula (18) is 2″S-7,8-dihydropyran-4′-hydroxy-5-methyl isoflavone; yellow amorphous powder, UV (ACN) 259nm; ECD (MeOH) 215 (Δε-2.96), 248 (Δ-0.42); through its high-resolution mass spectrum, the quasi-molecular ion peak [M+H] + m / z 369.1325 (calculated value 369.1260) confirmed its molecular formula to be C 21 H 21 O6; According to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2″S-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone. 1 H and 13 CNMR attribution see Table 2 [600MHz( 1 H), 150MHz( 13 C), CD3OD];

[0059] Compound (19) is 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone; yellow amorphous powder, UV (ACN) 210, 259, 299 nm; through its high-resolution mass spectrum quasi-molecular ion peak [MH] - m / z 287.1288 (calculated value 287.1362) confirmed its molecular formula to be C 17 H 19 O4; according to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone. 1 H and 13 C NMR assignments are shown in Table 3 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0060] Compound (20) is 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone; yellow amorphous powder, UV (ACN) 202, 259, 299 nm; through its high-resolution mass spectrum quasi-molecular ion peak [MH] - m / z 301.1443 (calculated value 301.1518) confirmed its molecular formula to be C 18 H 21 O4; according to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone. 1 H and13 C NMR assignments are shown in Table 3 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0061] Compound (21) is 6,7-pyran-2-isopropyl-5-hydroxychromone; yellow amorphous powder, UV (ACN) 218, 253, 337 nm; through its high-resolution mass spectrum, the quasi-molecular ion peak [M+H] + m / z 303.1226 (calculated value 303.1154) confirmed its molecular formula to be C 17 H 19 O5; according to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 6,7-pyran-2-isopropyl-5-hydroxychromone. 1 H and 13 C NMR assignments are shown in Table 3 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0062] Compound (22) is 2′S-8-geranyl-5,7,4′-trihydroxyflavone; yellow amorphous powder, UV (ACN) 293,355nm; through its high-resolution mass spectrum quasi-molecular ion peak [MH] - m / z 407.11856 (calculated value 407.1937) confirmed its molecular formula to be C 25 H 27 O5; according to 1 H, 13 Its structure was confirmed by C NMR and 2D NMR data and named as 2′S-8-geranyl-5,7,4′-trihydroxydihydroflavone; 1 H and 13 C NMR assignments are shown in Table 2 [600 MHz ( 1 H), 150MHz( 13 C), CD3OD];

[0063] Table 1. Compounds of Formula 1-8 1 H data [δ(ppm), J(Hz)]

[0064]

[0065] Table 2. Compounds of Formula 9, 10, 11, 12, 17, 18 and 22 1 H and 13 C NMR data [δ(ppm), J(Hz)]

[0066]

[0067]

[0068] Table 3. Compounds of Formulas 13-16, 19, 20 and 21 1 H and 13 C NMR data [δ(ppm), J(Hz)]

[0069] BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Figure 1 The compounds of formula (1) and (2) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0071] Figure 2 The compounds of formula (1) and (2) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0072] Figure 3 The measured and calculated circular dichroism spectra of the compounds of formula (1) and (2) of the present invention;

[0073] Figure 4 The compounds of formula (3) and (4) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0074] Figure 5 The compounds of formula (3) and (4) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0075] Figure 6 The measured and calculated circular dichroism spectra of the compounds of formula (3) and (4) of the present invention;

[0076] Figure 7 The compounds of formula (5) and (6) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0077] Figure 8 The compounds of formula (5) and (6) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0078] Fig. 9 The measured and calculated circular dichroism spectra of the compounds of formula (5) and (6) of the present invention;

[0079] Fig.10The compounds of formula (7) and (8) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0080] Fig.11 The compounds of formula (7) and (8) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0081] Fig.12 The measured and calculated circular dichroism spectra of the compounds of formula (7) and (8) of the present invention;

[0082] Fig.13 The compounds of formula (9) and (10) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0083] Fig.14 The compounds of formula (9) and (10) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0084] Fig.15 The measured and calculated circular dichroism spectra of the compounds of formula (9) and (10) of the present invention;

[0085] Fig.16 The compounds of formula (11) and (12) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0086] Fig.17 The compounds of formula (11) and (12) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0087] Fig.18 The measured and calculated circular dichroism spectra of the compounds of formula (11) and (12) of the present invention;

[0088] Fig.19 The compounds of formula (13) and (14) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0089] Fig. 20 The compounds of formula (13) and (14) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0090] Fig.21 The measured and calculated circular dichroism spectra of the compounds of formula (13) and (14) of the present invention;

[0091] Fig. 22The compounds of formula (15) and (16) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0092] Fig.23 The compounds of formula (15) and (16) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0093] Fig.24 The measured and calculated circular dichroism spectra of the compounds of formula (15) and (16) of the present invention;

[0094] Fig.25 The compounds of formula (17) and (18) of the present invention are 1 H NMR (600 MHz, CD3OD) spectrum;

[0095] Fig.26 The compounds of formula (17) and (18) of the present invention are 13 C NMR (150 MHz, CD3OD) spectrum;

[0096] Fig. 27 The measured and calculated circular dichroism spectra of the compounds of formula (17) and (18) of the present invention;

[0097] Fig.28 The compound of the present invention is 1 H NMR (600 MHz, CD3OD) spectrum;

[0098] Fig.29 The compound of the present invention is 13 C NMR (150 MHz, CD3OD) spectrum;

[0099] Fig.30 The compound of the present invention is of formula (20) 1 H NMR (600 MHz, CD3OD) spectrum;

[0100] Fig.31 The compound of the present invention is of formula (20) 13 C NMR (150 MHz, CD3OD) spectrum;

[0101] Fig.32 The compound of the present invention is 1 H NMR (600 MHz, CD3OD) spectrum;

[0102] Fig.33 The compound of the present invention is 13 C NMR (150 MHz, CD3OD) spectrum;

[0103] Fig.34 The compound of the present invention is 1 H NMR (600 MHz, CD3OD) spectrum;

[0104] Fig.35 The compound of the present invention is 13 C NMR (150 MHz, CD3OD) spectrum. DETAILED DESCRIPTION

[0105] All reagents used were analytical grade, and methanol and acetonitrile in HPLC were of HPLC grade (Merck, Germany). HPD-300 macroporous adsorption resin: produced by Beijing Solebow Technology Co., Ltd.; normal phase silica gel (100-200 mesh): produced by Qingdao Ocean Chemical Plant; Sephadex LH-20 gel: produced by General Electric Healthcare Group, USA; reversed phase silica gel ODS: produced by Merck, Germany; thin layer chromatography silica gel is HSGF254: produced by Yantai Huangwu Silica Gel Development and Pilot Plant; HPLC (Dionex, USA) was configured as follows: P680HPLC pump, ASI-100 autosampler, TCC-100 column oven, UVD170U ultraviolet detector (four wavelengths), quaternary solvent system, online degasser, Chameleon chromatography workstation. The preparative high performance liquid chromatography (Dionex, USA) was configured as follows: P680HPLC pump, UVD170U ultraviolet detector (four wavelengths), quaternary solvent system, online degasser, Chameleon chromatography workstation; mass spectra were measured using a quadrupole-time-of-flight hybrid mass spectrometer (Applied Biosystems, USA); nuclear magnetic resonance was measured using a VARIAN VNMRS 600 MHz nuclear magnetic resonance spectrometer; and ECD spectra were obtained using a Chirascan spectropolarimeter (UK).

[0106] Hops were collected from Changji City, Xinjiang Uygur Autonomous Region in October 20 and identified as female fruits of hops (Humulus lupulus L.) by Associate Researcher Lu Chunfang from the Xinjiang Institute of Physics and Chemistry, Chinese Academy of Sciences.

[0107] Example 1

[0108] a. Take 10 kg of hops, crush them, and then use 50 L of 99% ethanol aqueous solution at room temperature for cold extraction three times, and evaporate the solvent under reduced pressure to obtain a crude hop extract;

[0109] b. Dispersing the crude extract obtained in step a with water, adding petroleum ether, chloroform and n-butanol for extraction three times in sequence, combining the chloroform layers and evaporating under reduced pressure to obtain the chloroform layer extract;

[0110] c. Separate the chloroform extract obtained in step b by using a normal pressure macroporous adsorption resin column chromatography, and use a volume ratio of 10-100% methanol aqueous solution for gradient elution. The fractions are analyzed by silica gel thin layer chromatography, and the same fractions are combined to obtain 10 components Fr.1-Fr.10; (HPLC-DAD) ultraviolet tracking method is used to focus on separating the fractions with strong absorption at 370nm, and components Fr.5, Fr.6, Fr.7 was separated by medium pressure preparative chromatography, isocratically eluted with a methanol aqueous solution of 10-100% by volume, and the obtained sub-fractions were further separated by Sephadex LH-20 gel column chromatography with a methanol aqueous solution of 95% by volume, and by semi-preparative high performance liquid chromatography, with a methanol aqueous solution of 40-99% by volume, using a gradient elution to obtain the compound of formula (1): 2"R-2', 3'-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (2): 2"S-2', 3'-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (3): 2"S-3', 4'-dihydropyran-2', 4-dihydroxy-6-methyl chalcone; the compound of formula (4): 2"R-3', 4'-dihydropyran-2', 4-dihydroxy-6-methyl chalcone chalcone; the compound of formula (5) is: 2"S-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (6) is: 2"R-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (7) is: 2"S-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (8) is: 2"R-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (9) is: 2S, 2"R-7,8-dihydropyran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (10) is: 2S, 2"S-7,8-dihydropyran -4′-hydroxy-5-methyl dihydroflavone; the compound of formula (11) is: 2S, 2″R-7,8-dihydrofuran-4′-hydroxy-5-methyl dihydroflavone; the compound of formula (12) is: 2S, 2″S-7,8-dihydrofuran-4′-hydroxy-5-methyl dihydroflavone; the compound of formula (13) is: 2′S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (14) is: 2′R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (15) is: 2′S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone; the compound of formula (16) is: 2′R-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone The compound of formula (17) is: 2″R-7,8-dihydropyran-4′-hydroxy-5-methyl isoflavone; the compound of formula (18) is: 2″S-7,8-dihydropyran-4′-hydroxy-5-methyl isoflavone; the compound of formula (19) is: 6-isoprenyl-2-isopropyl-5,7-dihydroxy chromone; the compound of formula (20) is: 8-isoprenyl-2-sec-butyl-5,7-dihydroxy chromone; the compound of formula (21) is: 6,7-pyran-2-isopropyl-5-hydroxy chromone; the compound of formula (22) is: 2′S-8-geranyl-5,7,4′-trihydroxy dihydroflavone. .

[0111] Example 2

[0112] a. Take 10 kg of hops, crush them, and use 60 L of 50% ethanol aqueous solution, reflux extract at 80 ° C for 3 times, and evaporate the solvent under reduced pressure to obtain a crude hop extract;

[0113] b. Dispersing the crude extract obtained in step a with water, adding n-hexane, chloroform and n-butanol in sequence for extraction 4 times, combining the chloroform layers and evaporating under reduced pressure to obtain a chloroform extract;

[0114] c. Separate the chloroform extract obtained in step b by using a normal pressure macroporous adsorption resin column chromatography, and use a volume ratio of 10-100% ethanol aqueous solution for gradient elution. The fractions are analyzed by silica gel thin layer chromatography, and the same fractions are combined to obtain 10 components Fr.1-Fr.10; (HPLC-DAD) ultraviolet tracking method is used to focus on separating the fractions with strong absorption at 370nm, and components Fr.5, Fr.6, Fr.7 is separated by medium pressure preparative chromatography, gradient elution with acetonitrile-water solution of 10-100% by volume, and the obtained sub-fractions are further separated by Sephadex LH-20 gel column chromatography with 95% methanol-water, and by semi-preparative high performance liquid chromatography, gradient elution with acetonitrile-water solution of 30-99% by volume, to obtain the compound of formula (1): 2″R-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (2): 2″S-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (3): 2″S-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone; the compound of formula (4): 2″R-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone; The compound of formula (5) is: 2"S-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (6) is: 2"R-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (7) is: 2"S-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (8) is: 2"R-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (9) is: 2S, 2"R-7,8-dihydropyran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (10) is: 2S, 2"S-7,8-dihydropyran-4 '-hydroxy-5-methyl dihydroflavone; the compound of formula (11) is: 2S, 2"R-7,8-dihydrofuran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (12) is: 2S, 2"S-7,8-dihydrofuran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (13) is: 2'S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (14) is: 2'R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (15) is: 2'S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone; the compound of formula (16) is: 2'R-6,7-dihydro Pyran-2-sec-butyl-5-hydroxychromone; the compound of formula (17) is: 2″R-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (18) is: 2″S-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (19) is: 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone; the compound of formula (20) is: 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone; the compound of formula (21) is: 6,7-pyran-2-isopropyl-5-hydroxychromone; the compound of formula (22) is: 2′S-8-geranyl-5,7,4′-trihydroxydihydroflavone. .

[0115] Example 3

[0116] a. Take 10 kg of hops, crush them, and then percolate and extract them three times with 90 L of anhydrous ethanol at room temperature, and evaporate the solvent under reduced pressure to obtain a crude hop extract;

[0117] b. Dispersing the crude extract obtained in step a with water, sequentially adding n-hexane, chloroform and n-butanol for extraction 5 times, combining the chloroform layers and evaporating under reduced pressure to obtain a chloroform extract;

[0118] c. Separate the chloroform extract obtained in step b by using a normal pressure macroporous adsorption resin column chromatography, using a volume ratio of 10-100% methanol-water solution gradient elution, analyze the fractions by silica gel thin layer chromatography, combine the same fractions, and obtain 10 components Fr.1-Fr.10; use (HPLC-DAD) ultraviolet tracking method to focus on separating the fractions with strong absorption at 370nm, and separate components Fr.5, Fr.6, Fr.7 is separated by medium pressure preparative chromatography, gradient elution with acetonitrile aqueous solution at a volume concentration of 10-100%, and the obtained sub-fractions are further separated by Sephadex LH-20 gel column chromatography with 95% methanol-water, and by semi-preparative high performance liquid chromatography, gradient elution with methanol aqueous solution at a volume concentration of 40-99%, to obtain the compound of formula (1): 2″R-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (2): 2″S-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (3): 2″S-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone; the compound of formula (4): 2″R-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone; The compound of formula (5) is: 2"S-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (6) is: 2"R-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (7) is: 2"S-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (8) is: 2"R-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (9) is: 2S, 2"R-7,8-dihydropyran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (10) is: 2S, 2"S-7,8-dihydropyran-4 '-hydroxy-5-methyl dihydroflavone; the compound of formula (11) is: 2S, 2"R-7,8-dihydrofuran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (12) is: 2S, 2"S-7,8-dihydrofuran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (13) is: 2'S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (14) is: 2'R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (15) is: 2'S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone; the compound of formula (16) is: 2'R-6,7-dihydro Pyran-2-sec-butyl-5-hydroxychromone; the compound of formula (17) is: 2″R-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (18) is: 2″S-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (19) is: 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone; the compound of formula (20) is: 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone; the compound of formula (21) is: 6,7-pyran-2-isopropyl-5-hydroxychromone; the compound of formula (22) is: 2′S-8-geranyl-5,7,4′-trihydroxydihydroflavone. .

[0119] Example 4

[0120] a. Take 10 kg of hops, crush them, and then reflux extract them with 90 L of 70% methanol aqueous solution at 80° C. for 3 times, and evaporate the solvent under reduced pressure to obtain a crude hop extract;

[0121] b. Dispersing the crude extract obtained in step a with water, sequentially adding n-hexane, chloroform and n-butanol for extraction 4 times, combining the chloroform layers and evaporating under reduced pressure to obtain a chloroform extract;

[0122] c. Separate the chloroform extract obtained in step b by normal pressure macroporous adsorption resin column chromatography, and use a volume ratio of 10-100% ethanol-water solution gradient elution. The fractions are analyzed by silica gel thin layer chromatography, and the same fractions are combined to obtain 10 components Fr.1-Fr.10; (HPLC-DAD) ultraviolet tracking method is used to focus on the fractions with strong absorption at 370nm, and components Fr.5, Fr.6, Fr.7 is separated by medium pressure preparative chromatography, isocratically eluted with a methanol-water solution having a volume concentration of 10-100%, and the obtained sub-fractions are further separated by Sephadex LH-20 gel column chromatography with 95% methanol-water, and by preparative high performance liquid chromatography with an acetonitrile aqueous solution having a volume concentration of 30-99%, using a gradient elution to obtain the compound of formula (1): 2″R-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (2): 2″S-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (3): 2″S-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone; the compound of formula (4): 2″R-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone; The compound of formula (5) is: 2"S-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (6) is: 2"R-4', 5'-dihydrofuran-6', 4-dihydroxy-6-methyl chalcone; the compound of formula (7) is: 2"S-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (8) is: 2"R-2,3-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (9) is: 2S, 2"R-7,8-dihydropyran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (10) is: 2S, 2"S-7,8-dihydropyran-4 '-hydroxy-5-methyl dihydroflavone; the compound of formula (11) is: 2S, 2"R-7,8-dihydrofuran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (12) is: 2S, 2"S-7,8-dihydrofuran-4'-hydroxy-5-methyl dihydroflavone; the compound of formula (13) is: 2'S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (14) is: 2'R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (15) is: 2'S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone; the compound of formula (16) is: 2'R-6,7-dihydro Pyran-2-sec-butyl-5-hydroxychromone; the compound of formula (17) is: 2″R-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (18) is: 2″S-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (19) is: 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone; the compound of formula (20) is: 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone; the compound of formula (21) is: 6,7-pyran-2-isopropyl-5-hydroxychromone; the compound of formula (22) is: 2′S-8-geranyl-5,7,4′-trihydroxydihydroflavone. .

[0123] Example 5

[0124] a. Take 10 kg of hops, crush them, and extract them three times with 50 L of 99% cold soaking liquid, and evaporate the solvent under reduced pressure to obtain a crude hop extract;

[0125] b. Dispersing the crude extract obtained in step a with water, sequentially adding petroleum ether, chloroform and n-butanol for extraction 4 times, combining the chloroform layers and evaporating under reduced pressure to obtain a chloroform extract;

[0126] c. Separate the chloroform extract obtained in step b by pressurized macroporous adsorption resin column chromatography, and use a volume ratio of 10-100% ethanol-water gradient elution. The fractions are analyzed by silica gel thin layer chromatography (TLC), and the same fractions are combined to obtain 8 components (Fr.1-Fr.8); separate component Fr.4 by medium pressure preparative chromatography, and use a volume ratio of 10-100% acetonitrile-water gradient elution to obtain 6 components Fr.4.1-Fr.4.6; separate component Fr.4.3. The semi-preparative high performance liquid chromatography was used for separation, with a volume concentration of 30-99% acetonitrile aqueous solution, and gradient elution was used to obtain the compound of formula (1): 2"R-2', 3'-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (2): 2"S-2', 3'-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; the compound of formula (3): 2"S-3', 4'-dihydropyran-2', 4-dihydroxy-6-methyl chalcone; the compound of formula (4): 2"R-3', 4'-dihydropyran-2', 4-dihydroxy-6-methyl chalcone; the compound of formula (5): 2"S-4' , 5′-dihydrofuran-6′, 4-dihydroxy-6-methyl chalcone; the compound of formula (6) is: 2″R-4′, 5′-dihydrofuran-6′, 4-dihydroxy-6-methyl chalcone; the compound of formula (7) is: 2″S-2,3-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (8) is: 2″R-2,3-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (9) is: 2S, 2″R-7,8-dihydropyran-4′-hydroxy-5-methyl dihydroflavone; the compound of formula (10) is: 2S, 2″S-7,8-dihydropyran-4′-hydroxy-5-methyl dihydroflavone The compound of formula (11) is: 2S, 2″R-7, 8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone; the compound of formula (12) is: 2S, 2″S-7, 8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone; the compound of formula (13) is: 2′S-6, 7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (14) is: 2′R-6, 7-dihydropyran-2-isopropyl-5-hydroxychromone; the compound of formula (15) is: 2′S-6, 7-dihydropyran-2-sec-butyl-5-hydroxychromone; the compound of formula (16) is: 2′R-6, 7-dihydropyran-2- sec-butyl-5-hydroxychromone; the compound of formula (17) is: 2″R-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (18) is: 2″S-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; the compound of formula (19) is: 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone; the compound of formula (20) is: 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone; the compound of formula (21) is: 6,7-pyran-2-isopropyl-5-hydroxychromone; the compound of formula (22) is: 2′S-8-geranyl-5,7,4′-trihydroxydihydroflavone. .

[0127] Example 6

[0128] The use of any one of the xanthohumol derivative compounds isolated from hops in Examples 1-5 in the preparation of anti-tuberculosis drugs is described by taking unlabeled self-luminous Mycobacterium tuberculosis as an example:

[0129] Experimental methods:

[0130] The minimum inhibitory concentration of the compound was determined using unlabeled autoluminescent Mycobacterium tuberculosis (Autoluminescent H37Ra). The H37Ra strain was inoculated in 7H9 medium, and 200 μL of culture medium was taken for detection. When the relative light unit (RLU) reached 2 million, 25 μL of samples with different concentration gradients (4-64 μg / mL) were added. DMSO was used as a blank control and rifampicin (1 μg / ml) was used as a positive control. Each sample was repeated 3 times. The RLU value was detected after 7 days of administration. The minimum inhibitory concentration was defined as: RLU drug / RLU DMSO <10%, experimental results:

[0131] Table 4. Anti-Mycobacterium tuberculosis activity test results of compounds of formula (1) to formula (22)

[0132]

[0133] Note: MIC lux It was defined as the lowest concentration that inhibited greater than 90% RLU compared with the negative control (DMSO);

[0134] It can be seen from Table 3 that the compounds of formula (1) to formula (22) all have different degrees of anti-Mycobacterium tuberculosis activity and can be used to prepare anti-tuberculosis drugs.

Claims

1. A xanthohumol derivative in hops, characterized in that The structural formula of the derivative is: in: The compound of formula (1) is named: 2″R-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (2) is named: 2″S-2′, 3′-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (3) is named: 2″S-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone; the compound of formula (4) is named: 2″R-3′, 4′-dihydropyran-2′, 4-dihydroxy-6-methyl chalcone Ketone; the compound of formula (5) is named: 2″S-4′, 5′-dihydrofuran-6′, 4-dihydroxy-6-methyl chalcone; the compound of formula (6) is named: 2″R-4′, 5′-dihydrofuran-6′, 4-dihydroxy-6-methyl chalcone; the compound of formula (7) is named: 2″S-2,3-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; the compound of formula (8) is named: 2″R-2,3-dihydropyran-4′, 4-dihydroxy-6-methyl chalcone; The compound of formula (9) is named: 2S, 2″R-7, 8-dihydropyran-4′-hydroxy-5-methyldihydroflavone; The compound of formula (10) is named: 2S,2″S-7,8-dihydropyran-4′-hydroxy-5-methyldihydroflavone; The compound of formula (11) is named: 2S, 2″R-7, 8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone; The compound of formula (12) is named: 2S,2″S-7,8-dihydrofuran-4′-hydroxy-5-methyldihydroflavone; The compound of formula (13) is named: 2′S-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; The compound of formula (14) is named: 2′R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone; The compound of formula (15) is named: 2′S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone; The compound of formula (16) is named: 2′R-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone; The compound of formula (17) is named: 2″R-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; The compound of formula (18) is named: 2″S-7,8-dihydropyran-4′-hydroxy-5-methylisoflavone; The compound of formula (19) is named: 6-isoprenyl-2-isopropyl-5,7-dihydroxychromone; The compound of formula (20) is named: 8-isoprenyl-2-sec-butyl-5,7-dihydroxychromone; The compound of formula (21) is named: 6,7-pyran-2-isopropyl-5-hydroxychromone; The compound of formula (22) is named: 2'S-8-geranyl-5,7,4'-trihydroxydihydroflavone.

2. The method for preparing xanthohumol derivatives from hops according to claim 1, characterized in that Follow these steps: a. Take female hops as raw materials, dry and crush them, and then use 5-10 times the volume of 50-99% ethanol aqueous solution, anhydrous ethanol, 50-99% methanol aqueous solution or anhydrous methanol to perform percolation, cold soaking or heating reflux extraction at room temperature, and concentrate to obtain a crude extract of hops; b. Disperse the crude extract obtained in step a with water, add petroleum ether or n-hexane, chloroform, ethyl acetate, and n-butanol in sequence, extract for 3-5 times, and concentrate the chloroform extract to obtain a chloroform extract concentrate; c. Separating the chloroform extract obtained in step b by macroporous adsorption resin column chromatography, normal phase silica gel column chromatography, medium pressure preparative chromatography, Sephadex LH-20 gel column chromatography and semi-preparative high performance liquid chromatography to obtain compounds 1 to 22; The macroporous adsorption resin column chromatography method used is normal pressure or pressurized column chromatography, the filler used is macroporous adsorption resin, the eluent used is 10-90% methanol aqueous solution or ethanol aqueous solution, and isocratic elution or gradient elution is adopted; the normal phase silica gel column chromatography method used is normal pressure column chromatography, the eluent is n-hexane-ethyl acetate mixed solvent, and gradient elution is adopted; the medium pressure preparation chromatography method uses reverse phase C-18 filler, the eluent is methanol aqueous solution or acetonitrile aqueous solution with a volume concentration of 30-100%, and isocratic elution or gradient elution is adopted; the Sephadex LH-20 gel column chromatography method used is normal pressure column chromatography, the eluent is a mixture of methanol, chloroform and water solvents, and isocratic elution is adopted; the preparation high performance liquid chromatography method used is pressurized column chromatography, the eluent is methanol aqueous solution with a volume concentration of 40-99% or acetonitrile aqueous solution with a volume concentration of 30-99%, and isocratic elution or gradient elution is adopted.

3. Use of the xanthohumol derivatives in hops according to claim 1 in the preparation of anti-tuberculosis drugs.

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