A xanthohumol derivative in hops, a preparation method and use thereof

Abstract

The present application provides a kind of humulon derivative in hops and preparation method and use, the humulon derivative is from the female flower fruit of hops using thin layer chromatography and analytical high performance liquid chromatography detection analysis, with organic solvent extraction, then through solvent extraction method, macroporous adsorption resin column chromatography, normal phase silica gel column chromatography, medium pressure preparation chromatography, Sephadex LH-20 gel column chromatography, semi-preparative high performance liquid chromatography is separated, 22 new humulon derivatives are obtained, 23 monomers are determined as new humulon derivatives by high resolution mass spectrometry, nuclear magnetic resonance spectroscopy, circular dichroism spectrum and quantum chemistry calculation method, and its structure identification is carried out.In addition, the minimum concentration (MIC 90 ) of inhibiting Mycobacterium tuberculosis (Mtb) H37Ra is used to evaluate the inhibitory effect of humulon derivative on Mtb H37Ra strain, which can be used for preparing anti-tuberculosis drugs.

Description

Technical Field

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

[0002] Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is a chronic infectious disease that has long plagued humanity and remains 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 remained the second largest single infectious cause of death in the world, second only to the coronavirus, and the number of deaths caused was 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 address the serious problem of drug resistance.

[0003] Hops (Humulus lupulus L.), also known as hops (commonly known in English as hop), lupulus, and hops, are perennial climbing herbs belonging to the genus Humulus in the subfamily Cannabaceae. They are an essential ingredient in beer brewing, imparting its unique bitterness and aroma, while also preserving and clarifying wort. They are known as the "soul" of beer. Hops are also an important traditional medicinal ingredient. Native to Europe, they grow wild in Xinjiang, my country, and are cultivated in Northeast China, North China, and Shandong. Xinjiang, with its abundant sunlight and large temperature swings between day and night, is the primary distribution area for wild hops in my country. Its female cones are not only an additive in beer brewing but also widely used as a medicine worldwide. Hops have stomachic, digestive, diuretic, calming, cough-relieving, and expectorant properties. They are used to treat loss of appetite, abdominal distension, tuberculosis, pleurisy, insomnia, hysteria, edema, and cystitis. They are included in numerous traditional Chinese medicines and are a unique Xinjiang botanical resource used both as a medicine and a food. Among them, the first part 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 structure of isoprenyl chalcone (about 0.1% to 1% of hops dry weight). In fact, hops is the only source of natural isoprenyl flavonoids found so far, such as xanthohumol and isoxanthohumol, which exist in plants in the form of glycoside. Hops is 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 isoprenyl flavonoids. The hard resin component of hops, xanthohumol and its derivative isoxanthohumol, is 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 cholagogue, anti-ulcer, liver protection, spasmolysis, diuresis, anti-virus, etc.; isoprenyl flavonoids also have anti-proliferation and anti-cancer effects.

[0005] However, so far, there has been no report on the effect of xanthohumol derivatives in hops on Mycobacterium tuberculosis.

[0006] Among them, the published invention patent: a xanthohumol derivative and its preparation method and use (patent number: ZL2020 10957790.8) is the previous work of this subject, and the derivative is obtained by chemical synthesis and modification method using 1″, 2″-dihydroxanthohumol C, xanthohumol C and 1″, 2″-dihydroxanthohumol K as raw materials, a series of compounds are obtained and preliminary anti-inflammatory activity test is carried out. The present invention is based on the previous work, and the chemical components similar to the structure of xanthohumol in hops are tracked and separated according to the ultraviolet absorption characteristics to obtain 22 monomer compounds by using phytochemical method. SUMMARY

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

[0008] The xanthohumol derivative compound in hops provided by the present application has the following structural formula:

[0009]

[0010] wherein:

[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'-dihydropyran-6',4-dihydroxy-6- methylchalcone;

[0016] The compound of formula (6) is named: 2"R-4',5'-dihydropyran-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- methylflavanone;

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

[0021] The compound of formula (11) is named: 2S,2"R-7,8-dihydropyran-4'-hydroxy-5- methylflavanone;

[0022] The compound of formula (12) is named: 2S,2"S-7,8-dihydropyran-4'-hydroxy-5- methylflavanone;

[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 as 2'S-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone;

[0026] The compound of formula (16) is named as 2'R-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone;

[0027] The compound of formula (17) is named as 2"S-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone;

[0028] The compound of formula (18) is named as 2"S-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone;

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

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

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

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

[0033] The preparation method of the humulon derivative in the hops is carried out according to the following steps:

[0034] a. Taking the female flower fruit of hops as raw material, drying and crushing, and then carrying out percolation, cold soaking or heating reflux extraction at room temperature by using 5-10 times amount of volume concentration of 50-99% ethanol aqueous solution, anhydrous ethanol, volume concentration of 50-99% methanol aqueous solution or anhydrous methanol, and then concentrating to obtain the crude extract of hops;

[0035] b. Dispersing the crude extract obtained in step a with water, and then adding petroleum ether or n-hexane, chloroform and n-butanol for 3-5 times of extraction, respectively, and then concentrating the chloroform extract to obtain the chloroform extract infusion;

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

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

[0038] The use of the xanthohumol derivative in the beer in the preparation of an anti-tuberculosis drug.

[0039] The xanthohumol derivative in the beer according to the application is obtained by separation and purification from plants, and can also be obtained by chemical modification method.

[0040] The xanthohumol derivative compound in the beer according to the application is determined in structure by using high resolution mass spectrometry, one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy, circular dichroism spectroscopy and quantum chemical calculation method, and the structure identification process is as follows:

[0041] The compound formula (1) is 2''R-2', 3'-dihydropyran-4', 4-dihydroxy-6-methyl chalcone; yellow amorphous powder, UV (ACN) 332 nm; ECD (MeOH) 210 (Δε+22.3), 297 (Δε+6.09), 337 (Δε-3.18); the molecular formula is C + m / z 371.1488 (calculated value is 371.1416) is determined by the quasi-molecular ion peak [M+H] 21 H 23 O6; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data, the structure is determined, and named as 2''R-2', 3'-dihydropyran-4', 4-dihydroxy-6-methyl chalcone, which 1 H and 13 C NMR assignment is shown in table 1 [600MHz( 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 is + 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 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); the quasi-molecular ion peak [MH] in its high-resolution mass spectrum is - 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″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); the quasi-molecular ion peak [MH] in its high-resolution mass spectrum is -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); the quasi-molecular ion peak [MH] in its high-resolution mass spectrum is - 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 it was named 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); the quasi-molecular ion peak [MH] in its high-resolution mass spectrum is - 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 it was named as 2″R-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone. 1 H and 13C NMR assignments are given in Table 1 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (C), CD3OD]; 13 H), 150 MHz (C), CD3OD];

[0047] Compound of formula (7) is 2"S-2,3-dihydro- pyran-4',4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV(ACN) 372 nm; ECD(MeOH) 207 (Δε + 8.05), 270 (Δε + 3.24), 308 (Δε + 0.24), 362 (Δε - 1.89); its molecular formula is C - m / z 369.1314 (calcd 369.1416) determined its molecular formula as C 21 H 21 O6; according to 1 H, 13 C NMR and 2D NMR data, it was identified as 2"S-2,3-dihydro- pyran-4',4-dihydroxy-6-methylchalcone, which 1 H and 13 C NMR assignments are given in Table 1 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (C), CD3OD]; 13 H), 150 MHz (C), CD3OD];

[0048] Compound of formula (8) is 2"R-2,3-dihydro- pyran-4',4-dihydroxy-6-methylchalcone; yellow amorphous powder, UV(ACN) 372 nm; ECD(MeOH) 207 (Δε - 8.43), 270 (Δε + 3.26), 311 (Δε - 1.03), 363 (Δε + 1.18); its molecular formula is C - m / z 369.1314 (calcd 369.1416) determined its molecular formula as C 21 H 21 O6; according to 1 H, 13 C NMR and 2D NMR data, it was identified as 2"R-2,3-dihydro- pyran-4',4-dihydroxy-6-methylchalcone, which 1 H and 13 C NMR assignments are given in Table 1 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (C), CD3OD]; 13 H), 150 MHz (C), CD3OD];

[0049] Compound of formula (9) is 2S,2"R-7,8-dihydrochroman-4'-hydroxy-5- methyldihydroflavanone; yellow amorphous powder, UV (ACN) 217, 290 nm; ECD (MeOH) 217 (Δε + 17.20), 236 (Δε + 6.91), 289 (Δε - 27.2), 334 (Δε + 9.37); its structure was determined by high resolution mass spectrum of the quasi-molecular ion peak [M-H] - m / z 369.1331 (calculated 369.1416) to determine its molecular formula as C 21 H 22 O6; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data to determine its structure, named 2S,2"R-7,8-dihydrochroman-4'-hydroxy-5-methyldihydroflavanone, which 1 H and 13 C NMR assignment is shown in Table 2 [600MHz ( 1 H), 150MHz ( 13 C), CD3OD];

[0050] Compound of formula (10) is 2S,2"S-7,8-dihydrochroman-4'-hydroxy-5- methyldihydroflavanone; yellow amorphous powder, UV (ACN) 217, 290 nm; ECD (MeOH) 217 (Δε - 22.60), 236 (Δε - 11.10), 289 (Δε + 30.17), 335 (Δε - 11.26); its structure was determined by high resolution mass spectrum of the quasi-molecular ion peak [M-H] - m / z 369.1331 (calculated 369.1416) to determine its molecular formula as C 21 H 22 O6; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data to determine its structure, named 2S,2"S-7,8-dihydrochroman-4'-hydroxy-5-methyldihydroflavanone, which 1 H and 13 C NMR assignment is shown in Table 2 [600MHz ( 1 H), 150MHz ( 13 C), CD3OD];

[0051] Compound of formula (11) is 2S,2"R-7,8-dihydrofuran-4'-hydroxy-5- methyldihydroflavanone, yellow amorphous powder, UV(ACN) 218, 290 nm; ECD(MeOH) 214(Δε + 1.83), 239(Δε + 1.14), 289(Δε - 4.65), 326(Δε + 1.37); its structure was determined by high resolution mass spectrum of the quasi-molecular ion peak [M-H] - m / z 371.1488 (calculated value 371.1416) to determine its molecular formula as C 21 H 23 O6; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data to determine its structure, named 2S,2"R-7,8-dihydrofuran-4'-hydroxy-5-methyldihydroflavanone, which 1 H and 13 C NMR assignment is shown in Table 2 [600MHz ( 1 H), 150MHz ( 13 C), CD3OD];

[0052] Compound of formula (12) is 2S,2"S-7,8-dihydrofuran-4'-hydroxy-5- methyldihydroflavanone; yellow amorphous powder, UV(ACN) 218, 290 nm; ECD(MeOH) 215(Δε - 6.77), 236(Δε - 2.72), 291(Δε + 4.71), 327(Δε - 1.11); its structure was determined by high resolution mass spectrum of the quasi-molecular ion peak [M-H] - m / z 371.1488 (calculated value 371.1416) to determine its molecular formula as C 21 H 23 O6; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data to determine its structure, named 2S,2"S-7,8-dihydrofuran-4'-hydroxy-5-methyldihydroflavanone, which 1 H and 13 C NMR assignment is shown in Table 2 [600MHz ( 1 H), 150MHz ( 13 C), CD3OD];

[0053] Compound of 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); its structure was determined by high resolution mass spectrum of 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 (14) is 2′R-6,7-dihydropyran-2-isopropyl-5-hydroxychromone, a yellow amorphous powder, UV (ACN) 210, 251, 299 nm; ECD (MeOH) 212 (Δε-2.92), 256 (Δ+3.12), 285 (Δε-1.44); the quasi-molecular ion peak [M+H] in its high-resolution mass spectrum is + 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 (15) is 2′S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone, a yellow amorphous powder, UV (ACN) 210, 231, 251, 299 nm; ECD (MeOH) 210 (Δε+5.52), 230 (Δε+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 C NMR and two-dimensional nuclear magnetic resonance data confirmed its structure and named it 2′S-6,7-dihydropyran-2-sec-butyl-5-hydroxychromone. 1 H and 13C NMR assignments are given in Table 2 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (13C), CD3OD]; 13 C), CD3OD];

[0056] Compound of formula (16) is 2'R-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone, yellow amorphous powder, UV(ACN) 210, 231, 251, 299 nm; ECD(MeOH) 209 (Δε +5.52), 230 (Δε +3.17), 259 (Δε +3.06); its molecular formula is C + m / z 319.1541 (calcd 319.1467) determined its molecular formula as C 18 H 23 O5; according to 1 H, 13 C NMR and 2D NMR data, it was identified as 2'R-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone, which 1 H and 13 C NMR assignments are given in Table 3 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (13C), CD3OD]; 13 C), CD3OD];

[0057] Compound of formula (17) is 2"R-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone; yellow amorphous powder, UV(ACN) 259 nm; ECD(MeOH) 218 (Δε +4.58), 250 (Δε +1.35); its molecular formula is C + m / z 369.1325 (calcd 369.1260) determined its molecular formula as C 21 H 21 O6; according to 1 H, 13 C NMR and 2D NMR data, it was identified as 2"R-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone, which 1 H and 13 C NMR assignments are given in Table 2 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (13C), CD3OD]; 13 C), CD3OD];

[0058] Compound formula (18) is 2"S-7,8-dihydro-4'-hydroxy-5-methyl isoflavone; yellow amorphous powder, UV(ACN) 259 nm; ECD(MeOH) 215(Δε -2.96), 248(Δ -0.42); its molecular formula is determined as C + m / z 369.1325 (calcd 369.1260) to determine its molecular formula as C 21 H 21 O6; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data to determine its structure, named 2"S-7,8-dihydro-4'-hydroxy-5-methyl isoflavone, which 1 H and 13 C NMR assignment is shown in Table 2 [600MHz ( 1 H), 150MHz ( 13 C), CD3OD];

[0059] Compound formula (19) is 6-prenyl-2-isopropyl-5,7-dihydroxychromone; yellow amorphous powder, UV(ACN) 210, 259, 299 nm; its molecular formula is determined as C - m / z 287.1288 (calcd 287.1362) to determine its molecular formula as C 17 H 19 O4; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data to determine its structure, named 6-prenyl-2-isopropyl-5,7-dihydroxychromone, which 1 H and 13 C NMR assignment is shown in Table 3 [600MHz ( 1 H), 150MHz ( 13 C), CD3OD];

[0060] Compound formula (20) is 8-prenyl-2-sec-butyl-5,7-dihydroxychromone; yellow amorphous powder, UV(ACN) 202, 259, 299 nm; its molecular formula is determined as C - m / z 301.1443 (calcd 301.1518) to determine its molecular formula as C 18 H 21 O4; according to 1 H, 13 C NMR and two-dimensional nuclear magnetic resonance data to determine its structure, named 8-prenyl-2-sec-butyl-5,7-dihydroxychromone, which 1 H and13 C NMR assignments are given in Table 3 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (C), CD3OD]; 13 C), CD3OD];

[0061] Compound of formula (21) is 6,7-pyranyl-2-isopropyl-5-hydroxychromone; yellow amorphous powder, UV(ACN) 218, 253, 337 nm; its molecular formula is C + m / z 303.1226 (calcd 303.1154) determined its molecular formula as C 17 H 19 O5; according to 1 H, 13 C NMR and 2D NMR data, its structure was determined and named as 6,7-pyranyl-2-isopropyl-5-hydroxychromone; its 1 H and 13 C NMR assignments are given in Table 3 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (C), CD3OD]; 13 C), CD3OD];

[0062] Compound of formula (22) is 2'S-8-geranyl-5,7,4'-trihydroxyflavanone; yellow amorphous powder, UV(ACN) 293, 355 nm; its molecular formula is C - m / z 407.11856 (calcd 407.1937) determined its molecular formula as C 25 H 27 O5; according to 1 H, 13 C NMR and 2D NMR data, its structure was determined and named as 2'S-8-geranyl-5,7,4'-trihydroxyflavanone; its 1 H and 13 C NMR assignments are given in Table 2 [600 MHz (1H), 150 MHz (13C), CD3OD]; 1 H), 150 MHz (C), CD3OD]; 13 C), CD3OD];

[0063] Table 1.1H NMR data [δ (ppm), J (Hz)] of compounds of formula 1-8 1 H data [δ (ppm), J (Hz)]

[0064]

[0065] Table 2.1H and C NMR data [δ (ppm), J (Hz)] of 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. Formula 13-16, 19, 20 and 21 compounds of the present invention 1 H and 13 C NMR data [δ (ppm), J (Hz)]

[0069] BRIEF DESCRIPTION OF DRAWINGS

[0070] Figure 1 Circular dichroism spectra of the present compounds of formula (1) and (2) as measured and calculated 1 H NMR (600 MHz, CD3OD) spectra;

[0071] Figure 2 Circular dichroism spectra of the present compounds of formula (1) and (2) as measured and calculated 13 C NMR (150 MHz, CD3OD) spectra;

[0072] Figure 3 Circular dichroism spectra of the present compounds of formula (3) and (4) as measured and calculated

[0073] Figure 4 Circular dichroism spectra of the present compounds of formula (3) and (4) as measured and calculated 1 H NMR (600 MHz, CD3OD) spectra;

[0074] Figure 5 Circular dichroism spectra of the present compounds of formula (3) and (4) as measured and calculated 13 C NMR (150 MHz, CD3OD) spectra;

[0075] Figure 6 Circular dichroism spectra of the present compounds of formula (5) and (6) as measured and calculated

[0076] Figure 7 Circular dichroism spectra of the present compounds of formula (5) and (6) as measured and calculated 1 H NMR (600 MHz, CD3OD) spectra;

[0077] Figure 8 Circular dichroism spectra of the present compounds of formula (5) and (6) as measured and calculated 13 C NMR (150 MHz, CD3OD) spectra;

[0078] Figure 9 Circular dichroism spectra of the present compounds of formula (5) and (6) as measured and calculated

[0079] Figure 10for the compounds of the present invention of formula (7) and (8) 1 H NMR (600 MHz, CD3OD) spectra;

[0080] Figure 11 for the compounds of the present invention of formula (7) and (8) 13 C NMR (150 MHz, CD3OD) spectra;

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

[0082] Figure 13 for the compounds of the present invention of formula (9) and (10) 1 H NMR (600 MHz, CD3OD) spectra;

[0083] Figure 14 for the compounds of the present invention of formula (9) and (10) 13 C NMR (150 MHz, CD3OD) spectra;

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

[0085] Figure 16 for the compounds of the present invention of formula (11) and (12) 1 H NMR (600 MHz, CD3OD) spectra;

[0086] Figure 17 for the compounds of the present invention of formula (11) and (12) 13 C NMR (150 MHz, CD3OD) spectra;

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

[0088] Figure 19 for the compounds of the present invention of formula (13) and (14) 1 H NMR (600 MHz, CD3OD) spectra;

[0089] Figure 20 for the compounds of the present invention of formula (13) and (14) 13 C NMR (150 MHz, CD3OD) spectra;

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

[0091] Figure 22for the compounds of the present invention of formula (15) and (16) 1 H NMR (600 MHz, CD3OD) spectra;

[0092] Figure 23 for the compounds of the present invention of formula (15) and (16) 13 C NMR (150 MHz, CD3OD) spectra;

[0093] Figure 24 Circular dichroism spectra measured and calculated for the compounds of the present invention of formula (15) and (16);

[0094] Figure 25 for the compounds of the present invention of formula (17) and (18) 1 H NMR (600 MHz, CD3OD) spectra;

[0095] Figure 26 for the compounds of the present invention of formula (17) and (18) 13 C NMR (150 MHz, CD3OD) spectra;

[0096] Figure 27 Circular dichroism spectra measured and calculated for the compounds of the present invention of formula (17) and (18);

[0097] Figure 28 for the compounds of the present invention of formula (19) 1 H NMR (600 MHz, CD3OD) spectra;

[0098] Figure 29 for the compounds of the present invention of formula (19) 13 C NMR (150 MHz, CD3OD) spectra;

[0099] Figure 30 for the compounds of the present invention of formula (20) 1 H NMR (600 MHz, CD3OD) spectra;

[0100] Figure 31 for the compounds of the present invention of formula (20) 13 C NMR (150 MHz, CD3OD) spectra;

[0101] Figure 32 for the compounds of the present invention of formula (21) 1 H NMR (600 MHz, CD3OD) spectra;

[0102] Figure 33 for the compounds of the present invention of formula (21) 13 C NMR (150 MHz, CD3OD) spectra;

[0103] Figure 34 Compound of formula (22) of the present application 1 H NMR (600MHz, CD3OD) spectrum;

[0104] Figure 35 Compound of formula (22) of the present application 13 C NMR (150MHz, CD3OD) spectrum. DETAILED DESCRIPTION

[0105] All reagents used are of analytical purity, and the methanol and acetonitrile used in high performance liquid chromatography are of high performance liquid chromatography grade (Merck, Germany). HPD-300 macroporous adsorption resin: produced by Beijing Solabio Science and Technology Co., Ltd.; normal phase silica gel (100-200 mesh): produced by Qingdao Marine Chemical Factory; Sephadex LH-20 gel: produced by General Electric Medical Group, USA; reverse phase silica gel ODS: produced by Merck, Germany; thin layer chromatography silica gel is HSGF254: produced by Yantai Huangwu Silica Gel Development and Test Factory; high performance liquid chromatography (Dionex, USA) is configured as follows: P680 HPLC pump, ASI-100 automatic sampler, TCC-100 column oven, UVD170U ultraviolet detector (four wavelengths), four-component solvent system, online degassing machine, and chameleon chromatography workstation. Preparative high performance liquid chromatography (Dionex, USA) is configured as follows: P680 HPLC pump, UVD170U ultraviolet detector (four wavelengths), four-component solvent system, online degassing machine, and chameleon chromatography workstation; mass spectrometry is determined by quadrupole-time-of-flight hybrid mass spectrometer (Applied Biosystems, USA); nuclear magnetic resonance is determined by VARIAN VNMRS 600MHz nuclear magnetic resonance spectrometer; ECD spectra are obtained using a Chirascan spectropolarimeter (UK).

[0106] Humulus lupulus L. was collected from Changji City, Xinjiang Uygur Autonomous Region in October 20 years, and identified by Dr. Lu Chunfang, Xinjiang Institute of Physico-Chemical Technology, Chinese Academy of Sciences, as the female fruit of Humulus lupulus L.

[0107] Example 1

[0108] a. Take 10 kg of Humulus lupulus L., crush it, and then extract it three times at room temperature by cold soaking with 50 L of 99% ethanol aqueous solution. Dry the solvent under reduced pressure to obtain the crude extract of Humulus lupulus L.;

[0109] b. Disperse the crude extract obtained in step a with water, and then extract it three times with petroleum ether, chloroform, and n-butanol in sequence. Combine the chloroform layers and then dry them under reduced pressure to obtain the chloroform layer extract.

[0110] c. The extract of chloroform from step b was subjected to column chromatography on a macroporous resin under normal pressure. The column was eluted with a gradient of 10-100% methanol in water. The fractions were analyzed by silica gel thin layer chromatography, and the same fractions were combined to obtain 10 fractions, Fr. 1-Fr. 10. The fractions with strong absorption at 370 nm were separated by HPLC-DAD, and the fractions Fr. 5, Fr. 6, and Fr. 10 were further separated by HPLC-MS.7 The compounds of formula (1) is 2"R-2',3'-dihydropyranyl-4',4-dihydroxy-6- methylchalcone; the compound of formula (2) is 2"S-2',3'-dihydropyranyl-4',4-dihydroxy- 6-methylchalcone; the compound of formula (3) is 2"S-3',4'-dihydropyranyl-2',4-dihydroxy- 6-methylchalcone; the compound of formula (4) is 2"R-3',4'-dihydropyranyl-2',4-dihydroxy- 6-methylchalcone; the compound of formula (5) is 2"S-4',5'-dihydropyranyl-6',4-dihydroxy- 6-methylchalcone; the compound of formula (6) is 2"R-4',5'-dihydropyranyl-6',4-dihydroxy- 6-methylchalcone; the compound of formula (7) is 2"S-2,3-dihydropyranyl-4',4-dihydroxy- 6-methylchalcone; the compound of formula (8) is 2"R-2,3-dihydropyranyl-4',4-dihydroxy- 6-methylchalcone; the compound of formula (9) is 2S,2"R-7,8-dihydropyranyl-4'-hydroxy-5- methylflavanone; the compound of formula (10) is 2S,2"S-7,8-dihydropyranyl-4'-hydroxy- 5-methylflavanone; the compound of formula (11) is 2S,2"R-7,8-dihydropyranyl-4'-hydroxy- 5-methylflavanone; the compound of formula (12) is 2S,2"S-7,8-dihydropyranyl-4'-hydroxy- 5-methylflavanone; the compound of formula (13) is 2'S-6,7-dihydropyranyl-2-isopropyl-5- hydroxychromone; the compound of formula (14) is 2'R-6,7-dihydropyranyl-2-isopropyl- 5-hydroxychromone; the compound of formula (15) is 2'S-6,7-dihydropyranyl-2-sec- butyl-5-hydroxychromone; the compound of formula (16) is 2'R-6,7-dihydropyranyl-2- sec-butyl-5-hydroxychromone; the compound of formula (17) is 2"R-7,8-dihydropyranyl- 4'-hydroxy-5-methylisoflavone; the compound of formula (18) is 2"S-7,8-dihydropyranyl- 4'-hydroxy-5-methylisoflavone; the compound of formula (19) is 6-prenyl-2-isopropyl-5,7- dihydroxychromone; the compound of formula (20) is 8-prenyl-2-sec-butyl-5,7- dihydroxychromone; the compound of formula (21) is 6,7-pyranyl-2-isopropyl-5- hydroxychromone; the compound of formula (22) is 2'S-8-geranyl-5,7,4'-trihydroxyflavanone.

[0111] Example 2

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

[0113] b. Disperse the crude extract obtained in step a with water, and successively extract it 4 times with n-hexane, chloroform, and n-butanol. Combine the chloroform layers and dry them under reduced pressure to obtain a chloroform extract infusion;

[0114] c. The extract of chloroform from step b was subjected to column chromatography on a macroporous resin under normal pressure. The column was eluted with a gradient of 10-100% ethanol in water. The fractions were analyzed by silica gel thin layer chromatography, and the same fractions were combined. Ten fractions were obtained, Fr. 1-Fr. 10. The fractions with strong absorption at 370 nm were separated by HPLC-DAD, and the fractions Fr. 5, Fr. 6, and Fr. 10 were further separated by HPLC-MS.7The compounds of formula (1) is: 2"R-2',3'-dihydropyranyl-4',4-dihydroxy-6- methylchalcone; the compound of formula (2) is: 2"S-2',3'-dihydropyranyl-4',4- dihydroxy-6-methylchalcone; the compound of formula (3) is: 2"S-3',4'-dihydropyranyl- 2',4-dihydroxy-6-methylchalcone; the compound of formula (4) is: 2"R-3',4'- dihydropyranyl-2',4-dihydroxy-6-methylchalcone; the compound of formula (5) is: 2"S-4',5'-dihydropyranyl-6',4-dihydroxy-6-methylchalcone; the compound of formula (6) is: 2"R-4',5'-dihydropyranyl-6',4-dihydroxy-6-methylchalcone; the compound of formula (7) is: 2"S-2,3-dihydropyranyl-4',4-dihydroxy-6-methylchalcone; the compound of formula (8) is: 2"R-2,3-dihydropyranyl-4',4-dihydroxy-6-methylchalcone; the compound of formula (9) is: 2S,2"R-7,8-dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compound of formula (10) is: 2S,2"S-7,8-dihydropyranyl-4'-hydroxy-5- methyldihydroflavone; the compound of formula (11) is: 2S,2"R-7,8-dihydropyranyl- 4'-hydroxy-5-methyldihydroflavone; the compound of formula (12) is: 2S,2"S-7,8- dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compound of formula (13) is: 2'S-6,7-dihydropyranyl-2-isopropyl-5-hydroxychromone; the compound of formula (14) is: 2'R-6,7-dihydropyranyl-2-isopropyl-5-hydroxychromone; the compound of formula (15) is: 2'S-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone; the compound of formula (16) is: 2'R-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone; the compound of formula (17) is: 2"R-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone; the compound of formula (18) is: 2"S-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone; the compound of formula (19) is: 6-prenyl-2-isopropyl-5,7-dihydroxychromone; the compound of formula (20) is: 8-prenyl-2-sec-butyl-5,7-dihydroxychromone; the compound of formula (21) is: 6,7-pyranyl-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 and extract 3 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. Disperse the crude extract obtained in step a with water, and sequentially extract 5 times with n-hexane, chloroform and n-butanol, combine the chloroform layers and evaporate the solvent under reduced pressure to obtain a chloroform extract infusion;

[0118] c. The extract of chloroform from step b was subjected to column chromatography on a macroporous resin under normal pressure. The column was eluted with a gradient of 10-100% methanol-water solution. The fractions were analyzed by silica gel thin layer chromatography. The same fractions were combined to obtain 10 fractions, Fr. 1-Fr. 10. The fractions with strong absorption at 370 nm were separated by HPLC-DAD. The fractions Fr. 5, Fr. 6, Fr. 7, Fr. 8, Fr. 9 and Fr. 10 were further separated by HPLC-MS.7The compounds of formula (1) is: 2"R-2',3'-dihydropyranyl-4',4-dihydroxy-6- methylchalcone; the compound of formula (2) is: 2"S-2',3'-dihydropyranyl-4',4- dihydroxy-6-methylchalcone; the compound of formula (3) is: 2"S-3',4'-dihydropyranyl- 2',4-dihydroxy-6-methylchalcone; the compound of formula (4) is: 2"R-3',4'- dihydropyranyl-2',4-dihydroxy-6-methylchalcone; the compound of formula (5) is: 2"S-4',5'-dihydropyranyl-6',4-dihydroxy-6-methylchalcone; the compound of formula (6) is: 2"R-4',5'-dihydropyranyl-6',4-dihydroxy-6-methylchalcone; the compound of formula (7) is: 2"S-2,3-dihydropyranyl-4',4-dihydroxy-6-methylchalcone; the compound of formula (8) is: 2"R-2,3-dihydropyranyl-4',4-dihydroxy-6-methylchalcone; the compound of formula (9) is: 2S,2"R-7,8-dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compound of formula (10) is: 2S,2"S-7,8-dihydropyranyl-4'-hydroxy-5- methyldihydroflavone; the compound of formula (11) is: 2S,2"R-7,8-dihydropyranyl- 4'-hydroxy-5-methyldihydroflavone; the compound of formula (12) is: 2S,2"S-7,8- dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compound of formula (13) is: 2'S-6,7-dihydropyranyl-2-isopropyl-5-hydroxychromone; the compound of formula (14) is: 2'R-6,7-dihydropyranyl-2-isopropyl-5-hydroxychromone; the compound of formula (15) is: 2'S-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone; the compound of formula (16) is: 2'R-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone; the compound of formula (17) is: 2"R-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone; the compound of formula (18) is: 2"S-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone; the compound of formula (19) is: 6-prenyl-2-isopropyl-5,7-dihydroxychromone; the compound of formula (20) is: 8-prenyl-2-sec-butyl-5,7-dihydroxychromone; the compound of formula (21) is: 6,7-pyranyl-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 extract them 3 times with 90 L of 70% methanol solution at 80°C under reflux. Dry the solvent under reduced pressure to obtain the hop crude extract;

[0121] b. Disperse the hop crude extract obtained in step a with water, and successively extract it 4 times with n-hexane, chloroform, and n-butanol. Combine the chloroform layers and dry them under reduced pressure to obtain the chloroform extract;

[0122] c. The extract of chloroform from step b was subjected to column chromatography on a macroporous resin under normal pressure. The column was eluted with a gradient of 10-100% ethanol-water solution. The fractions were analyzed by silica gel thin layer chromatography. The same fractions were combined to obtain 10 fractions, Fr. 1-Fr. 10. The fractions with strong absorption at 370 nm were separated by HPLC-DAD. The fractions Fr. 5, Fr. 6, Fr. 7, Fr. 8, Fr. 9 and Fr. 10 were further separated by HPLC-MS.7 The compounds of formula (1) is 2"R-2',3'-dihydropyranyl-4',4-dihydroxy-6- methylchalcone; the compound of formula (2) is 2"S-2',3'-dihydropyranyl-4',4-dihydroxy- 6-methylchalcone; the compound of formula (3) is 2"S-3',4'-dihydropyranyl-2',4-dihydroxy- 6-methylchalcone; the compound of formula (4) is 2"R-3',4'-dihydropyranyl-2',4-dihydroxy- 6-methylchalcone; the compound of formula (5) is 2"S-4',5'-dihydropyranyl-6',4-dihydroxy- 6-methylchalcone; the compound of formula (6) is 2"R-4',5'-dihydropyranyl-6',4-dihydroxy- 6-methylchalcone; the compound of formula (7) is 2"S-2,3-dihydropyranyl-4',4-dihydroxy- 6-methylchalcone; the compound of formula (8) is 2"R-2,3-dihydropyranyl-4',4-dihydroxy- 6-methylchalcone; the compound of formula (9) is 2S,2"R-7,8-dihydropyranyl-4'-hydroxy-5- methylflavanone; the compound of formula (10) is 2S,2"S-7,8-dihydropyranyl-4'-hydroxy- 5-methylflavanone; the compound of formula (11) is 2S,2"R-7,8-dihydropyranyl-4'-hydroxy- 5-methylflavanone; the compound of formula (12) is 2S,2"S-7,8-dihydropyranyl-4'-hydroxy- 5-methylflavanone; the compound of formula (13) is 2'S-6,7-dihydropyranyl-2-isopropyl-5- hydroxychromone; the compound of formula (14) is 2'R-6,7-dihydropyranyl-2-isopropyl- 5-hydroxychromone; the compound of formula (15) is 2'S-6,7-dihydropyranyl-2-sec- butyl-5-hydroxychromone; the compound of formula (16) is 2'R-6,7-dihydropyranyl-2- sec-butyl-5-hydroxychromone; the compound of formula (17) is 2"R-7,8-dihydropyranyl- 4'-hydroxy-5-methylisoflavone; the compound of formula (18) is 2"S-7,8-dihydropyranyl- 4'-hydroxy-5-methylisoflavone; the compound of formula (19) is 6-prenyl-2-isopropyl-5,7- dihydroxychromone; the compound of formula (20) is 8-prenyl-2-sec-butyl-5,7- dihydroxychromone; the compound of formula (21) is 6,7-pyranyl-2-isopropyl-5- hydroxychromone; the compound of formula (22) is 2'S-8-geranyl-5,7,4'-trihydroxyflavanone.

[0123] Example 5

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

[0125] b. Disperse the crude extract obtained in step a with water, and successively extract it four times with petroleum ether, chloroform, and n-butanol. Combine the chloroform layers and dry them by evaporation under reduced pressure to obtain a chloroform extract;

[0126] c. The extract of chloroform from step b was subjected to column chromatography on pressurized macroporous adsorption resin, eluted with gradient of 10-100% ethanol in water, and the fractions were analyzed by silica gel thin layer chromatography (TLC). The same fractions were combined to give 8 fractions (Fr. 1-Fr. 8). Fraction Fr. 4 was subjected to medium pressure preparative chromatography, eluted with gradient of 10-100% acetonitrile in water, to give 6 fractions Fr. 4.1-Fr. 4.6. Fraction Fr. 4.2 was subjected to medium pressure preparative chromatography, eluted with gradient of 10-100% acetonitrile in water, to give 2 fractions Fr. 4.2.1 and Fr. 4.2.2. Fraction Fr. 4.2.1 was subjected to medium pressure preparative chromatography, eluted with gradient of 10-100% acetonitrile in water, to give 2 fractions Fr. 4.2.1.1 and Fr. 4.2.1.2. Fraction Fr. 4.2.1.1 was subjected to medium pressure preparative chromatography, eluted with gradient of 10-100% acetonitrile in water, to give 2 fractions Fr. 4.2.1.1.1 and3The compounds of formula (1) are: 2"R-2',3'-dihydropyranyl-4',4-dihydroxy-6- methylchalcone; the compounds of formula (2) are: 2"S-2',3'-dihydropyranyl-4',4- dihydroxy-6-methylchalcone; the compounds of formula (3) are: 2"S-3',4'-dihydropyranyl- 2',4-dihydroxy-6-methylchalcone; the compounds of formula (4) are: 2"R-3',4'- dihydropyranyl-2',4-dihydroxy-6-methylchalcone; the compounds of formula (5) are: 2"S-4',5'-dihydropyranyl-6',4-dihydroxy-6-methylchalcone; the compounds of formula (6) are: 2"R-4',5'-dihydropyranyl-6',4-dihydroxy-6-methylchalcone; the compounds of formula (7) are: 2"S-2,3-dihydropyranyl-4',4-dihydroxy-6-methylchalcone; the compounds of formula (8) are: 2"R-2,3-dihydropyranyl-4',4-dihydroxy-6-methylchalcone; the compounds of formula (9) are: 2S,2"R-7,8-dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compounds of formula (10) are: 2S,2"S-7,8-dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compounds of formula (11) are: 2S,2"R-7,8-dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compounds of formula (12) are: 2S,2"S-7,8-dihydropyranyl-4'-hydroxy-5-methyldihydroflavone; the compounds of formula (13) are: 2'S-6,7-dihydropyranyl-2-isopropyl-5-hydroxychromone; the compounds of formula (14) are: 2'R-6,7-dihydropyranyl-2-isopropyl-5-hydroxychromone; the compounds of formula (15) are: 2'S-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone; the compounds of formula (16) are: 2'R-6,7-dihydropyranyl-2-sec-butyl-5-hydroxychromone; the compounds of formula (17) are: 2"R-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone; the compounds of formula (18) are: 2"S-7,8-dihydropyranyl-4'-hydroxy-5-methylisoflavone; the compounds of formula (19) are: 6-prenyl-2-isopropyl-5,7-dihydroxychromone; the compounds of formula (20) are: 8-prenyl-2-sec-butyl-5,7-dihydroxychromone; the compounds of formula (21) are: 6,7-pyranyl-2-isopropyl-5-hydroxychromone; the compounds of formula (22) are: 2'S-8-geranyl-5,7,4'-trihydroxydihydroflavone.

[0127] Example 6

[0128] Use of the xanthohumol derivative compound isolated from hops in any one of Examples 1-5 in the preparation of an anti-tuberculosis drug, taking the example of autoluminescent Mycobacterium tuberculosis:

[0129] Experimental method:

[0130] Autoluminescent H37Ra was used to determine the minimum inhibitory concentration of the compound, the H37Ra strain was inoculated in 7H9 medium, 200 μL of culture solution was detected, when the relative light unit (RLU) reached 2 million, 25 μL of sample with different concentration gradients (4-64 μg / mL) was 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, and 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. Test results of compounds of formula (1) to formula (22) for anti-tuberculosis activity

[0132]

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

[0134] As can be seen from Table 3, the compounds of formula (1) to formula (22) all have different degrees of anti-tuberculosis activity and can be used for the preparation of 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-methylchalcone; The compound of formula (2) is named: 2″S-2′,3′-dihydropyran-4′,4-dihydroxy-6-methylchalcone; The compound of formula (3) is named: 2″S-3′,4′-dihydropyran-2′,4-dihydroxy-6-methylchalcone; The compound of formula (4) is named: 2″R-3′,4′-dihydropyran-2′,4-dihydroxy-6-methylchalcone; The compound of formula (5) is named: 2″S-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone; The compound of formula (6) is named: 2″R-4′,5′-dihydrofuran-6′,4-dihydroxy-6-methylchalcone; The compound of formula (7) is named: 2″S-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone; The compound of formula (8) is named: 2″R-2,3-dihydropyran-4′,4-dihydroxy-6-methylchalcone; The compound of formula (10) is named: 2S,2″S-7,8-dihydropyran-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.

2. The method for preparing xanthohumol derivatives from hops according to claim 1, characterized in that Follow these steps: a. Taking female hops as raw material, drying and crushing them, and then performing percolation, cold soaking or heating reflux extraction at room temperature with 5-10 times the volume concentration of 50-99% ethanol aqueous solution, anhydrous ethanol, 50-99% methanol aqueous solution or anhydrous methanol, and concentrating to obtain a crude hop extract; b. Dispersing the crude extract obtained in step a with water, sequentially adding petroleum ether or n-hexane, chloroform, and n-butanol for extraction 3-5 times, concentrating the chloroform extract to obtain a chloroform extract concentrate; c. Separate 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 1-8, compound 10, and compounds 13-21; Among them, the macroporous adsorption resin column chromatography 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 used is normal pressure column chromatography, the eluent is n-hexane-ethyl acetate mixed solvent, and gradient elution is adopted; the medium pressure preparative chromatography used uses reversed phase C-18 filler, the eluent is 30-100% methanol aqueous solution or acetonitrile aqueous solution by volume concentration, and isocratic elution or gradient elution is adopted; the Sephadex LH-20 gel column chromatography used is normal pressure column chromatography, the eluent is a mixture of methanol, chloroform and water solvents, and isocratic elution is adopted; the preparative high performance liquid chromatography used is pressurized column chromatography, the eluent is 40-99% methanol aqueous solution or 30-99% acetonitrile aqueous solution by volume concentration, and isocratic elution or gradient elution is adopted.

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

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