An ent-6,7-closed-labdane-type oridonin hydroxyl-modified derivative, a preparation method and application thereof

Abstract

The application discloses an ent-6,7-closed ormosaone-type oridonin hydroxyl modification derivative, a preparation method and application thereof, the derivative is an oridonin derivative with a modified A-ring and hydroxyl groups at positions 6, 7 and 14, has a structure shown in general formula I, and an optical active body or a racemate, a diastereoisomer mixture or a pharmaceutically acceptable salt thereof; the derivative has stronger antitumor activity, can significantly inhibit the proliferation of various malignant tumor cells such as breast cancer and liver cancer, and has good drug stability and solubility, low toxic and side effects, and has a very good clinical application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of oridonin hydroxyl modification derivatives, especially to a kind of ent-6,7-closed ring kaurene type oridonin hydroxyl modification derivatives, also relates to the preparation method and application of the above-mentioned oridonin hydroxyl modification derivatives. BACKGROUND

[0002] Oridonin belongs to enanti-kaurene type tetracyclic diterpene, is one of the main active ingredients of Rabdosia plant winter grass, with antibacterial, anti-inflammatory, antitumor and other physiological activities.Oridonin has been used for the treatment of various cancers, and has good inhibitory activity on pancreatic cancer, lung cancer, colorectal cancer and other tumor cells.

[0003] Research found that oridonin mainly through inducing cell cycle arrest, apoptosis, autophagy and its cross effect to play anticancer activity.The unique α-methylene cyclopentanone structure is confirmed as the core mechanism of anti-tumor and antibacterial activity, and the inhibition of tumor has been verified in in vitro and in vivo experiments, and shows excellent biological activity;In addition, the unique α-methylene cyclopentanone structure is confirmed as the core of anti-tumor and antibacterial activity, and the inhibition of tumor has been verified in in vitro and in vivo experiments, and shows excellent biological activity;In vitro experiment results show that oridonin has good inhibitory effect on human esophageal squamous cell carcinoma, sl80, Ehrlich ascites carcinoma, human hepatoma BEL-7402, reticulum sarcoma solid type, lung cancer SPCA-1, breast cancer and leukemia and other tumors.However, due to the poor water solubility of oridonin, unknown target and poor selectivity, its clinical application as an antitumor drug is severely limited, and new drug compounds with stronger antitumor activity and lower toxicity are urgently needed. SUMMARY

[0004] The purpose of the present application is to provide an ent-6,7-closed ring kaurene type oridonin hydroxyl modification derivative with more excellent inhibitory activity on various tumor cells, and to provide the preparation method of the derivative and its application in antitumor drugs.

[0005] Technical scheme: The present application discloses an ent-6,7-closed ring kaurene type oridonin hydroxyl modification derivative, which is an oridonin derivative with A-ring and hydroxyl modification at positions 6,7 and 14, and the structure is shown as formula I:

[0006]

[0007] Wherein: R is selected from hydrogen atom, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C 6-10aryl, C 3-8 cycloalkyl, oxirane, aziridine, thiirane, oxetane, tetrahydrofuran, tetrahydropyrrole, pyridine, pyrimidine, piperidine, morpholine, pyrrole, furan, imidazole, pyrazole, quinoline, indole, benzofuran;

[0008] or R is substituted by 1 to 4 identical or different R1: said R1is selected from the group consisting of hydrogen atom, halogen, hydroxyl, cyano, nitro, carboxyl, -NH2, sulfonyl, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylamino; said C1-C6alkyl, C1-C6alkoxy, C1-C6alkylamino is substituted by 1 or 2 or 3 substituents selected from the group consisting of halogen, hydroxyl, cyano, C1-C6alkyl, C3-C7cycloalkyl, C2-C6alkenyl, C2-C6alkynyl or phenyl;

[0009] n is 1, 2 or 3.

[0010] wherein said derivative is preferably: R is selected from the group consisting of C1-C6alkyl, C1-C6alkoxy, C 6-10 aryl, C 3-8 cycloalkyl, aziridine, thiirane, oxetane, tetrahydrofuran, pyridine, pyrimidine, piperidine, morpholine, pyrrole, furan, imidazole, pyrazole, quinoline, indole, benzofuran; or R is substituted by 1 to 4 identical or different R1: R1is selected from the group consisting of hydrogen atom, halogen, nitro, -NH2, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylamino.

[0011] wherein said derivative is further preferably: R is selected from the group consisting of C1-C6alkyl, C1-C6alkoxy, C 6-10 aryl, C 3-8 cycloalkyl, aziridine, thiirane, oxetane, pyridine, pyrimidine, furan, imidazole, pyrazole, quinoline, indole, benzofuran; or R is substituted by 1 to 4 identical or different R1: R1is selected from the group consisting of halogen, nitro, -NH2, C1-C6alkoxy, C1-C6alkylamino; n is 1 or 2.

[0012] wherein said derivative is any one of the following structures:

[0013]

[0014]

[0015] wherein said derivative also includes its optically active forms or racemates, diastereomeric mixtures, or pharmaceutically acceptable salts thereof.

[0016] The preparation method of the derivative comprises the following steps:

[0017] (1) Dissolve oridonin in anhydrous acetone, add 2,2-dimethoxypropane and p-toluenesulfonic acid, and reflux to react, cool to room temperature, extract, wash, dry, and concentrate to obtain compound II;

[0018] (2) Dissolve compound II in dichloromethane, add triethylamine, and drop methylsulfonyl chloride to react, cool to room temperature, extract, wash, dry, and concentrate to obtain compound III;

[0019] (3) Dissolve compound III in DMF, add lithium carbonate and lithium bromide to react, cool to room temperature, filter, wash, concentrate, spin dry, and quickly pass compound IV through a column of dichloromethane;

[0020] (4) Dissolve compound IV in tetrahydrofuran, add hydrochloric acid solution to react, spin dry, wash with water, concentrate, and perform column chromatography to obtain compound V;

[0021] (5) Dissolve compound V in dichloromethane, react at room temperature, add a small amount of water to quench the reaction after the reaction is completed, extract, wash, dry, filter, vacuum concentrate, and perform column chromatography to obtain compound VI;

[0022] (6) Dissolve compound VI and formaldehyde in a mixed solution of toluene and ethanol, add p-toluenesulfonic acid and heat to reflux to react, or dissolve compound VI in ethylene glycol dimethyl ether, add NaOH and methyl chloroacetate and heat to reflux to react; after the reaction is completed, vacuum concentrate, and perform column chromatography to obtain the ent-6,7-closed ring kaurene type oridonin hydroxyl modification derivative, and the reaction general formula is as follows:

[0023]

[0024] The application further discloses a pharmaceutical composition comprising the derivative, and an optically active body or racemate, a diastereoisomer mixture, or a pharmaceutically acceptable salt or mixture of the derivative, and a pharmaceutically acceptable auxiliary or carrier.

[0025] The ent-6,7-closed ring kaurene type oridonin hydroxyl modification derivative or the pharmaceutical composition can also be applied to preparation of an antitumor drug.

[0026] The tumor comprises triple-negative breast cancer, leukemia, colon cancer or liver cancer.

[0027] The application is that the ent-6,7-closed ring kaurene type oridonin hydroxyl modification derivative exerts anticancer activity by inducing cell cycle arrest, cell apoptosis, cell autophagy and cross effects thereof.

[0028] Definitions of terms used in connection with the present application: Unless otherwise indicated, the initial definition of a group or term provided herein is intended to be in effect through the entire specification of this patent; for terms not specifically defined herein, the meaning given to the term by persons of ordinary skill in the art in light of the disclosure and context should prevail. The term "substituted" means that a hydrogen atom in a molecule is replaced by a substituent or substituents.

[0029] The term "halogen" means a fluorine (F), chlorine (Cl), bromine (Br), iodine (I), or astatine (At) ion.

[0030] The term "alkyl" denotes a straight-chained or branched saturated hydrocarbon group having the stated number of carbon atoms. The term "C1-C6 alkyl" includes, but is not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, neopentyl, n-hexyl, i-hexyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and the like. 16 The term "fatty chain" means a straight-chained or branched saturated hydrocarbon group having 1 to 6 carbon atoms. C1-C6 alkyl includes, but is not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, neopentyl, n-hexyl, i-hexyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and the like.

[0031] The term "alkenyl" denotes a straight-chained, branched, or cyclic, non-aromatic hydrocarbon group having a main chain of 2 to 6 carbon atoms and at least one carbon-carbon double bond. Thus, "C2-C6 alkenyl" denotes an alkenyl group having 2 to 6 carbon atoms in the main chain. Alkenyl groups include ethenyl, propenyl, butenyl, 2-methylbutenyl, and cyclohexenyl, and the like. The straight-chained, branched, or cyclic portion of the alkenyl group can contain the double bond and, if a substituted alkenyl group is specified, this portion can be substituted.

[0032] The term "alkynyl" denotes a straight-chained, branched, or cyclic, non-aromatic hydrocarbon group having a main chain of 2 to 6 carbon atoms and at least one carbon-carbon triple bond. Thus, "C2-C6 alkynyl" denotes an alkynyl group having 2 to 6 carbon atoms in the main chain. Alkynyl groups include ethynyl, propynyl, butynyl, 3-methylbutynyl, and the like. The straight-chained, branched, or cyclic portion of the alkynyl group can contain the triple bond and, if a substituted alkynyl group is specified, this portion can be substituted.

[0033] The term "heterocycle" denotes a saturated cyclic group of 3 to 6 ring atoms, one or two of which are heteroatoms selected from N, O, or S(O) m (wherein m is an integer from 0 to 2) and the remaining ring atoms are C, wherein one or two of the C atoms can optionally be replaced by a carbonyl group. The ring of the heterocycle group can optionally be independently substituted by one, two, or three substituents.

[0034] The term "aromatic ring", "aryl" denotes a fully carbon monocyclic or fused polycyclic group of 6 to 10 carbon atoms, having a fully conjugated pi-electron system. Non-limiting examples of aromatic rings or aryl groups are phenyl, naphthyl, and anthryl.

[0035] The term "heteroaromatic ring", "heteroaryl" means a monocyclic or fused ring group of 5 to 10 ring atoms, containing one, two, three or four ring heteroatoms selected from N, O or S, with the remaining ring atoms being C, and additionally having a fully conjugated pi-electron system. Non-limiting examples of unsubstituted heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine and carbazole. The heteroaromatic ring or heteroaryl group can be substituted or unsubstituted. When substituted, the substituents are preferably one or more, more preferably one, two or three, yet more preferably one or two.

[0036] Invention principle: The ent-6,7-closed ring kaurene type oridonin hydroxyl modified derivatives of the present application are A-ring and 6,7,14 position hydroxyl modified oridonin derivatives. By sulfonyl modification of the 14 position hydroxyl, two hydrogen bond acceptors in the sulfonyl structure increase the binding force between the compound and the receptor. The 6,7 position hydroxyl is protected by cyclization technology to improve the liposolubility of the compound, achieve the enhancement of the antitumor activity, and reduce the toxicity to normal cells. The present application improves and optimizes the structure of oridonin, designs and synthesizes a series of ent-6,7-closed ring kaurene type oridonin hydroxyl modified derivatives with more excellent inhibitory activity on various tumor cells.

[0037] Beneficial effects: Compared with the prior art, the present application has the following remarkable advantages: (1) The ent-6,7-closed ring kaurene type oridonin hydroxyl modified derivatives of the present application have stronger antitumor activity, which can significantly inhibit the proliferation of various malignant tumor cells such as breast cancer and liver cancer; (2) The derivatives of the present application also have good pharmacokinetic stability and solubility, low toxicity and side effects, and great clinical application prospect. DETAILED DESCRIPTION

[0038] The technical solutions of the present application are further described below in combination with examples. The test materials used in the examples can be purchased through conventional channels.

[0039] Example 1

[0040] The compound 1 of the present application is shown as follows:

[0041]

[0042] The compound 1 of the present application can be synthesized according to the following reaction route:

[0043]

[0044] (1) Preparation of compound (II): oridonin (3 g, 8.23 mmol) was dissolved in 40 mL of anhydrous acetone, 2,2-dimethoxypropane 3 mL and a catalytic amount of p-toluenesulfonic acid (150 mg) were added, dry tube protection, reflux reaction for 10 min, the solution changed from white turbidity to clear, and 3 mL of 2,2-dimethoxypropane was added again, TLC monitoring reaction until the raw material disappeared, cooling to room temperature, saturated NaHCO3 solution was added, dichloromethane was extracted three times, 30 mL each time, saturated brine was washed twice, anhydrous sodium sulfate was dried, concentrated to get 3.17 g of white solid (yield 95%), which can be directly used for the next step reaction. 1 H NMR (500 MHz, CDCI3) δ 5.69 (d, J = 3.3 Hz, 2H), 4.12 (d, J = 11.0 Hz, 1H), 4.02 (d, J = 11.2 Hz, 1H), 3.84 (dd, J = 6.8, 5.1 Hz, 1H), 3.42-3.36 (m, 2H), 3.31-3.23 (m, 1H), 3.03 (d, J = 5.1 Hz, 1H), 2.47-2.37 (m, 1H), 2.32 (dddd, J = 8.1, 4.7, 3.3, 2.3 Hz, 1H), 1.77-1.71 (m, 1H), 1.74-1.66 (m, 1H), 1.70-1.63 (m, 1H), 1.66-1.57 (m, 2H), 1.60-1.49 (m, 1H), 1.51-1.42 (m, 1H), 1.40-1.30 (m, 1H), 1.00 (s, 5H), 0.94 (ddd, J = 6.7, 3.8, 2.6 Hz, 1H); 13 C NMR (125 MHz, CDCI3) δ 206.53, 153.58, 118.42, 100.94, 98.20, 74.35, 73.46, 72.30, 65.65, 61.19, 60.68, 50.88, 46.64, 41.35, 34.75, 34.15, 29.80, 29.04, 28.00, 25.83; ESI-MS m / z: 405.2272 calcd for C 23 H 33 O6[M+H] + 405.2278.

[0045] (2) Preparation of compound (III): The product of step 1 was dissolved in 30 mL of pre-dried dichloromethane, 2 mL of triethylamine was added, stirred in ice bath, 2 mL of methanesulfonyl chloride was added dropwise within 15 min, continued to stir for 1 h, TLC detected the disappearance of raw materials, added 80 mL of dichloromethane, washed twice with water, once with saturated brine, 20 mL each time, dried over anhydrous sodium sulfate, concentrated to give a light yellow solid 3 g (yield 79.4%). 1 H NMR (500 MHz, CDCI3) δ 5.69 (d, J = 3.3 Hz, 2H), 4.54 (dt, J = 6.4, 3.8 Hz, 1H), 4.12 (d, J = 11.0 Hz, 1H), 4.02 (d, J = 11.2 Hz, 1H), 3.84 (dd, J = 7.0, 5.1 Hz, 1H), 3.39 (dd, J = 4.6, 3.7 Hz, 1H), 3.03 (d, J = 5.1 Hz, 1H), 2.98 (s, 2H), 2.47 - 2.37 (m, 1H), 2.32 (dddd, J = 8.1, 4.7, 3.3, 2.3 Hz, 1H), 1.93 - 1.83 (m, 1H), 1.77 - 1.71 (m, 1H), 1.74 - 1.63 (m, 1H), 1.55 (s, 5H), 1.66 - 1.42 (m, 3H), 1.35 (ddd, J = 12.6, 9.3, 7.3 Hz, 1H), 1.00 (s, 4H), 0.94 (ddd, J = 7.0, 3.7, 2.4 Hz, 1H); 13 C NMR (125 MHz, CDCI3) δ 206.53, 153.58, 118.42, 100.94, 98.20, 81.40, 74.35, 72.30, 65.71, 60.77, 60.68, 50.22, 46.64, 40.60, 38.84, 36.70, 34.15, 29.80, 29.04, 26.60, 25.83, 25.54, 19.95; ESI-MS m / z: 483.2047 calcd for C 24 H 35 O8S[M+H] + 483.2042.

[0046] (3) Preparation of compound (IV): Compound (III) (3 g, 6.22 mmol) was dissolved in 25 mL of pre-dried DMF, lithium carbonate (4.59 g, 62.16 mmol), lithium bromide (5.4 g, 62.16 mmol) were added, reacted at 110 °C for 1 h, cooled to room temperature, filtered to remove lithium carbonate, lithium bromide with a sand funnel, the solid was washed with dichloromethane three times, the organic layers were combined, dichloromethane was added to 120 mL, washed with water twice, saturated brine once, washed with DMF to remove each time 30 mL, concentrated, the residual DMF was spin-dried with an oil pump, and the light yellow solid 2.08 g (yield 86.7%) was obtained by rapid column chromatography with dichloromethane. 1 H NMR (500 MHz, CDC13) δ 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 3.84 (dd, J = 7.0, 4.8 Hz, 1H), 3.42 - 3.35 (m, 2H), 3.03 (d, J = 4.8 Hz, 1H), 2.76 (dd, J = 10.2, 0.9 Hz, 1H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.2 Hz, 1H), 2.32 (dddd, J = 8.1, 4.7, 3.3, 2.3 Hz, 1H), 1.84 (ddd, J = 15.2, 5.1, 0.9 Hz, 1H), 1.68 - 1.57 (m, 2H), 1.60 - 1.49 (m, 1H), 1.39 (dddt, J = 7.7, 6.0, 4.0, 1.8 Hz, 1H), 1.38 - 1.28 (m, 1H), 1.07 (ddd, J = 7.0, 2.6, 1.8 Hz, 1H), 1.02 (s, 5H); 13 CNMR (125 MHz, CDC13) δ 206.53, 153.58, 131.28, 129.05, 118.42, 100.94, 98.20, 74.35, 71.57, 68.94, 60.75, 59.13, 49.37, 46.64, 41.44, 40.11, 34.58, 31.73, 29.80, 25.54, 23.17, 19.77; ESI-MS m / z: 387.2166 calcd for C 23 H 31 O5[M+H] + 387.2173.

[0047] (4) Preparation of compound (V): Compound (IV) (2.08 g, 5.38 mmol) was dissolved in 35 mL of tetrahydrofuran, 10 mL of 10% hydrochloric acid solution was added, stirred at room temperature for 1.5 h, the tetrahydrofuran was rotary evaporated, dichloromethane was added to 100 mL, washed with water three times, 25 mL each time, washed with saturated brine once, 25 mL each time, concentrated, and column chromatography was performed with dichloromethane:methanol = 100:1 to obtain 1.657 g (84.4% yield) of white solid. 1 H NMR (500 MHz, CDC13) δ 6.39 (d, J = 4.6 Hz, 1H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 1.8 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.92 (ddd, J = 5.7, 4.6, 3.7 Hz, 1H), 4.31 (s, 1H), 3.64 (dd, J = 8.2, 4.9 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 3.03 (d, J = 5.1 Hz, 1H), 3.02 - 2.96 (m, 1H), 2.76 (dd, J = 10.2, 0.9 Hz, 1H), 2.42 (dddd, J = 13.0, 8.3, 5.8, 2.5 Hz, 1H), 1.84 (ddd, J = 15.2, 5.1, 0.9 Hz, 1H), 1.68 - 1.49 (m, 2H), 1.39 (dddd, J = 6.4, 5.5, 2.7, 1.8 Hz, 1H), 1.38 - 1.28 (m, 1H), 1.07 (ddd, J = 8.2, 2.6, 1.8 Hz, 1H), 1.02 (s, 4H); 13 C NMR (125 MHz, CDC13) δ 208.70, 153.06, 131.28, 129.05, 119.64, 100.58, 73.25, 72.94, 66.78, 61.56, 60.13, 51.80, 44.95, 41.44, 40.11, 34.58, 31.73, 30.27, 23.17, 18.02; ESI-MS m / z: 347.1853 calcd for C 20 H 37 O5[M+H] + 347.1846.

[0048] (5) Preparation of compound (VI): Compound (V) (150 mg, 0.43 mmol) was dissolved in 10 mL of dichloromethane and reacted at room temperature for 30 min. The reaction was completed by TLC detection of the disappearance of the starting material. The reaction was quenched by adding a small amount of water, extracted three times with dichloromethane, washed with saturated brine, dried and filtered, concentrated under vacuum, and column chromatography was used to obtain 84.4 mg of pale yellow solid (yield 40%). 1 H NMR (500MHz, CDCl3) δ8.91(t,J=1.6Hz,1H),8.84(ddd,J=3.8,2.1,1.4Hz,1H),8.34(dt,J=8.2,2.0Hz,1H),7.59(dd,J=8.3,4.0Hz,1H),5.88(dtt, J=8.2,1.8,0.9Hz,1H),5.69(d,J=3.3Hz,2H),5.36(dt,J=8.1,4.9Hz,1H) ,4.89(dd,J=6.7,3.8Hz,1H),4.31(s,1H),3.64(dd,J=8.2,4.9Hz,1H),3. 38(dd,J=10.2,0.9Hz,1H),3.03(d,J=5.1Hz,1H),2.83-2.73(m,2H),2.42 (dddd,J=12.8,8.4,5.9,2.6Hz,1H),1.84(dddd,J=15.2,5.1,0.9Hz,1H),1 .68-1.49(m,3H),1.39(dddd,J=5.7,4.1,3.4,2.3Hz,1H),1.33(dddd,J=1 3.4,8.9,5.9,3.0Hz,1H),1.07(ddd,J=8.2,2.6,1.8Hz,1H),1.02(s,5H); 13 C NMR (125MHz, CDCl3) δ208.75,153.77,149.71,149.58,142.75,134.46,131.28,129.05,124.91,118.42,100. 54,83.61,72.94,66.78,61.83,60.13,51.46,46.02,41.44,40.11,34.58,31.73,30.27,23.17,18.02; ESI-MS m / z:488.1743calcd for C 25 H 30 NO7S[M+H] + 488.1750.

[0049] (6) Preparation of compound 1: Compound (VI) (30 mg, 0.06 mmol) and formaldehyde (2.5 mL, 0.07 mmol) were dissolved in a mixed solution of toluene and ethanol, a catalytic amount of p-toluenesulfonic acid (5 mg) was added, heated to reflux at 90 °C for 1.5 h, concentrated in vacuo, and column chromatography gave 20 mg of yellow solid (yield 65%). 1 H NMR (500 MHz, CDC13) δ 8.93-8.89 (m, 1H), 8.84 (ddd, J = 3.8, 2.1, 1.4 Hz, 1H), 8.34 (dt, J = 8.2, 2.0 Hz, 1H), 7.59 (dd, J = 8.3, 4.0 Hz, 1H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 5.24 (dd, J = 3.0, 1.7 Hz, 1H), 5.15 (d, J = 2.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.54 (dd, J = 7.3, 1.8 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83-2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88-1.80 (m, 1H), 1.68-1.49 (m, 3H), 1.43-1.24 (m, 3H), 1.02 (s, 4H); 13 C NMR (125 MHz, CDC13) δ 207.39, 153.58, 149.71, 149.58, 142.75, 134.46, 131.28, 129.05, 124.91, 118.42, 108.75, 92.92, 84.20, 76.93, 68.94, 60.34, 53.38, 48.51, 47.08, 42.12, 40.11, 34.53, 31.73, 29.80, 23.17, 19.77; ESI-MS m / z: 500.1737 calcd for C 27 H 32 N6O2 + [M+H] + 500.1744.

[0050] Example 2

[0051] The compound 2 of the present application and the preparation method are shown as follows:

[0052]

[0053] Reference is made to the synthesis method of compound 1 in Example 1.1 H NMR (500 MHz, CDC13) δ 7.50 (s, 1H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 5.24 (dd, J = 3.0, 1.7 Hz, 1H), 5.15 (d, J = 2.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 4.03 (s, 3H), 3.54 (dd, J = 7.3, 1.8 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.55 (m, 2H), 1.58 - 1.49 (m, 1H), 1.43 - 1.24 (m, 3H), 1.02 (s, 5H); 13 CNMR (125 MHz, CDC13) δ 207.39, 153.58, 134.79, 131.28, 129.05, 124.30, 120.31, 118.42, 108.75, 92.92, 85.55, 76.93, 68.94, 60.34, 53.38, 48.51, 47.08, 42.12, 40.11, 38.86, 34.53, 31.73, 29.80, 23.17, 19.77; ; ESI-MS m / z: 503.1846 calcd for C 25 H 31 N2O7S + [M+H] + 503.1839.

[0054] Example 3

[0055] The compound 3 of the present application and the preparation method are shown as follows:

[0056]

[0057] Reference to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 5.24 (d, J = 3.1 Hz, 1H), 5.15 (d, J = 2.9 Hz, 1H), 4.89 (dd, J = 7.0, 3.8 Hz, 1H), 3.54 (d, J = 6.2 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 3.01 (t, J = 8.5 Hz, 2H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.94 (d, J = 8.6 Hz, 1H), 1.93 - 1.80 (m, 2H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.10 (t, J = 8.6 Hz, 3H), 1.02 (s, 5H); 13 C NMR (125 MHz, CDC13) δ 206.53, 153.77, 130.65, 127.51, 118.42, 108.75, 92.92, 89.81, 79.49, 67.86, 60.34, 54.35, 49.64, 48.51, 46.02, 42.12, 40.81, 32.91, 30.27, 27.78, 18.54, 17.85, 13.09; ESI-MS m / z: 465.1498 calcd for C 27 H 32 N6O2[M+H] + 465.1505.

[0058] Example 4

[0059] The compound 4 of the present application and the preparation method are shown as follows:

[0060]

[0061] Reference to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 7.42 - 7.34 (m, 3H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 4.33 - 4.27 (m, 3H), 3.64 (dd, J = 8.2, 4.9 Hz, 1H), 3.38 (dd, J = 10.3, 0.9 Hz, 1H), 3.03 (d, J = 5.1 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.49 (m, 3H), 1.42 - 1.36 (m, 1H), 1.33 (dddd, J = 13.4, 8.8, 5.9, 3.0 Hz, 1H), 1.07 (ddd, J = 8.2, 2.6, 1.8 Hz, 1H), 1.02 (s, 5H); 13 C NMR (125 MHz, CDC13) δ 208.75, 153.77, 131.54, 131.28, 131.09, 129.41, 129.05, 128.19, 118.42, 100.54, 86.68, 72.94, 66.78, 61.83, 60.13, 58.00, 51.46, 46.02, 41.44, 40.11, 34.58, 31.73, 30.27, 23.17, 18.02; ESI-MS m / z: 501.1942 calcd for C 27 H 33 O7S + [M+H] + 501.1939.

[0062] Example 5

[0063] The compound 5 of the present application and the preparation method are shown as follows:

[0064]

[0065] Reference is made to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 8.79 (dd, J = 4.1, 1.7 Hz, 1H), 8.43 (d, J = 2.0 Hz, 1H), 8.10 (dd, J = 7.6, 1.8 Hz, 1H), 8.06 - 7.96 (m, 2H), 7.48 (dd, J = 7.6, 4.1 Hz, 1H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.02 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.2, 0.9 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.51 - 2.44 (m, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01 - 1.94 (m, 1H), 1.96 - 1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.18, 150.62, 149.62, 146.94, 139.15, 136.44, 136.09, 131.75, 127.94, 126.88, 125.89, 125.11, 122.68, 118.30, 103.63, 96.10, 83.22, 81.22, 66.32, 57.62, 51.66, 43.92, 43.29, 41.30, 40.81, 32.93, 31.24, 30.38, 28.55, 19.21; ESI-MS m / z: 550.1894 calcd for C 30 H 32 NO7S + [M+H] + 550.1901.

[0066] Example 6

[0067] The compound 6 of the present application and the preparation method are shown as follows:

[0068]

[0069] The synthesis method of compound 1 in Example 1 was referred to. 1 H NMR (500 MHz, CDC13) δ 7.86-7.80 (m, 2H), 7.71-7.64 (m, 2H), 5.74-5.65 (m, 2H), 5.49-5.40 (m, 2H), 5.02 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.3, 0.9 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.6, 2.9 Hz, 1H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01-1.94 (m, 1H), 1.96-1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78-1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.18, 149.62, 139.15, 135.88, 133.33, 130.00, 129.71, 125.89, 118.30, 103.63, 96.10, 83.21, 81.22, 66.32, 57.62, 51.66, 43.92, 43.29, 41.30, 40.81, 32.93, 31.24, 30.38, 28.55, 19.21; ESI-MS m / z: 577.0890 calcd for C 27 H 30 BrO7S + [M+H] + 577.0894.

[0070] Example 7

[0071] The compound 7 of the present application and the preparation method are shown as follows:

[0072]

[0073] The synthesis method of compound 1 in Example 1 was referred to. 1H NMR (500 MHz, CDC13) δ 7.68 - 7.61 (m, 3H), 7.63 - 7.54 (m, 2H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.02 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.2, 0.9 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01 - 1.94 (m, 1H), 1.96 - 1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H);13C NMR (125 MHz, CDC13) δ 202.18, 149.62, 139.15, 136.75, 134.13, 129.29, 128.66, 125.89, 118.30, 103.63, 96.10, 83.21, 81.22, 66.32, 57.62, 51.66, 43.92, 43.29, 41.30, 40.81, 32.93, 31.24, 30.38, 28.55, 19.21; ESI-MS m / z: 499.1785 calcd for C 27 H 31 O7S + [M+H] + 499.1791。

[0074] Example 8

[0075] The compound 8 of the present application and the preparation method are shown as follows:

[0076]

[0077] Reference is made to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 5.74-5.65 (m, 2H), 5.49-5.40 (m, 2H), 4.97 (dd, J = 6.8, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.2, 0.9 Hz, 1H), 3.34-3.26 (m, 1H), 3.30-3.24 (m, 1H), 2.51-2.44 (m, 1H), 2.10-1.81 (m, 8H), 1.80-1.63 (m, 5H), 1.56 (ddq, J = 12.3, 8.9, 6.3 Hz, 2H), 1.51-1.42 (m, 2H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.08, 149.53, 139.15, 125.89, 118.30, 104.19, 96.10, 81.27, 80.27, 66.32, 63.25, 57.33, 51.66, 43.89, 43.43, 41.30, 40.81, 32.93, 31.30, 30.38, 28.55, 27.42, 26.52, 25.57, 19.21; ESI-MS m / z: 505.2255 calcd for C 27 H 37 O7S + [M+H] + 505.2258.

[0078] Example 9

[0079] The compound 9 of the present application and the preparation method are shown as follows:

[0080]

[0081] Reference is made to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 8.05 - 7.98 (m, 2H), 7.93 - 7.87 (m, 2H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.02 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.91 (s, 2H), 3.80 (dd, J = 10.2, 0.9 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01 - 1.94 (m, 1H), 1.96 - 1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.18, 166.23, 149.62, 140.79, 139.15, 135.00, 130.55, 128.62, 125.89, 118.30, 103.63, 96.10, 83.21, 81.22, 66.32, 57.62, 52.05, 51.66, 43.92, 43.29, 41.30, 40.81, 32.93, 31.24, 30.38, 28.55, 19.21; ESI-MS m / z: 557.1840 calcd for C 29 H 33 O9S + [M+H] + 557.1844.

[0082] Example 10

[0083] The compound 10 of the present application and the preparation method are shown as follows:

[0084]

[0085] Reference to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 5.74-5.65 (m, 2H), 5.49-5.40 (m, 2H), 4.97 (dd, J = 6.8, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.3, 0.9 Hz, 1H), 3.39-3.24 (m, 2H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.7, 3.1 Hz, 1H), 2.00-1.93 (m, 1H), 1.96-1.91 (m, 2H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78-1.67 (m, 2H), 1.46 (dd, J = 11.2, 7.9 Hz, 2H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.08, 149.53, 139.15, 125.89, 118.30, 104.19, 96.10, 81.27, 80.24, 66.32, 57.33, 51.66, 43.89, 43.43, 41.30, 40.81, 37.35, 32.93, 31.30, 30.38, 28.55, 19.21, 10.29; ESI-MS m / z: 463.1785 calcd for C 24 H 31 O7S + [M+H] + 463.1789.

[0086] Example 11

[0087] The compound 11 of the present application and the preparation method are shown as follows:

[0088]

[0089] Reference to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 7.86-7.80 (m, 2H), 7.45-7.38 (m, 2H), 5.74-5.65 (m, 2H), 5.49-5.40 (m, 2H), 5.02 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.3, 0.9 Hz, 1H), 3.65-3.59 (m, 4H), 3.32 (dddd, J = 6.8, 5.8, 4.6, 2.9 Hz, 1H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01-1.94 (m, 1H), 1.96-1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78-1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.18, 172.64, 149.62, 139.15, 138.79, 135.12, 130.52, 127.90, 125.89, 118.30, 103.63, 96.10, 83.21, 81.22, 66.32, 57.62, 51.90, 51.66, 43.92, 43.29, 41.30, 40.81, 40.62, 32.93, 31.24, 30.38, 28.55, 19.21; ESI-MS m / z: 571.1996 calcd for C 30 H 35 O9S + [M+H] + 571.1998.

[0090] Example 12

[0091] The compound 12 of the present application and the preparation method are shown as follows:

[0092]

[0093] Reference is made to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 7.44 (d, J = 3.1 Hz, 1H), 6.99 (dd, J = 5.5, 1.6 Hz, 1H), 6.82 (dd, J = 5.5, 1.3 Hz, 1H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.11 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.3, 0.9 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01 - 1.94 (m, 1H), 1.96 - 1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 201.93, 149.65, 146.94, 144.73, 139.15, 125.89, 118.30, 115.25, 113.56, 104.19, 96.10, 82.19, 81.22, 66.32, 58.12, 51.66, 43.93, 43.31, 41.30, 40.81, 32.93, 31.27, 30.38, 28.55, 19.21; ESI-MS m / z: 489.1578 calcd for C 25 H 29 O8S + [M+H] + 489.1580.

[0094] Example 13

[0095] The compound 13 of the present application and the preparation method are shown as follows:

[0096]

[0097] Reference is made to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 7.31 - 7.23 (m, 3H), 7.18 (ddddt, J = 4.9, 4.0, 3.1, 1.9, 1.0 Hz, 2H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.04 (dd, J = 7.0, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.3, 1.0 Hz, 1H), 3.42 - 3.24 (m, 3H), 3.01 (ddt, J = 11.1, 10.2, 0.9 Hz, 2H), 2.51 - 2.44 (m, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.7, 3.1 Hz, 1H), 2.00 - 1.94 (m, 1H), 1.96 - 1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.8 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.08, 149.53, 139.15, 135.37, 129.63, 128.70, 127.16, 125.89, 118.30, 104.21, 96.10, 81.61, 81.27, 66.32, 57.25, 54.08, 51.66, 43.89, 43.46, 41.30, 40.81, 32.93, 31.26, 31.00, 30.38, 28.55, 19.21; ESI-MS m / z: 527.2098 calcd for C 29 H 35 O7S + [M+H] + 527.2091.

[0098] Example 14

[0099] The compound 14 of the present application and the preparation method are shown as follows:

[0100]

[0101] Reference is made to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 9.58 (s, 1H), 7.84-7.78 (m, 2H), 7.69-7.63 (m, 2H), 5.74-5.65 (m, 2H), 5.49-5.40 (m, 2H), 5.02 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.2, 0.9 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.6, 2.9 Hz, 1H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01-1.94 (m, 1H), 1.96-1.91 (m, 1H), 1.92 (d, J = 0.9 Hz, 1H), 1.91 (d, J = 0.7 Hz, 1H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.73 (ddt, J = 12.9, 8.6, 5.5 Hz, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.18, 169.62, 149.62, 143.62, 139.15, 130.38, 129.43, 125.89, 120.89, 118.30, 103.63, 96.10, 83.21, 81.22, 66.32, 57.62, 51.66, 43.92, 43.29, 41.30, 40.81, 32.93, 31.24, 30.38, 28.55, 24.01, 19.21; ESI-MS m / z: 556.2000 calcd for C 29 H 34 NO8S + [M+H] + 556.2004.

[0102] Example 15

[0103] The compound 15 of the present application and the preparation method are shown as follows:

[0104]

[0105] Reference is made to the synthesis method of compound 1 in Example 1. 1H NMR (500 MHz, CDC13) δ 8.77 (d, J = 6.8 Hz, 1H), 8.19 (t, J = 2.1 Hz, 1H), 7.74 (dd, J = 7.8, 2.1 Hz, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.22 - 7.17 (m, 1H), 6.64 (dd, J = 3.2, 2.2 Hz, 1H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.02 (dd, J = 6.9, 3.8 Hz, 1H), 4.84 (d, J = 2.9 Hz, 1H), 4.77 (dd, J = 2.9, 1.8 Hz, 1H), 4.64 (dd, J = 7.4, 1.7 Hz, 1H), 4.12 (dd, J = 10.4, 0.9 Hz, 1H), 3.80 (dd, J = 10.3, 0.9 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.47 (tdd, J = 5.7, 2.8, 1.7 Hz, 1H), 2.05 (dddd, J = 13.4, 8.8, 5.8, 3.1 Hz, 1H), 2.01 - 1.93 (m, 1H), 1.96 - 1.90 (m, 3H), 1.86 (ddt, J = 13.4, 8.6, 5.9 Hz, 1H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.18, 149.62, 139.15, 138.62, 130.84, 129.59, 125.89, 125.75, 123.58, 121.32, 118.30, 111.86, 103.63, 103.13, 96.10, 83.22, 81.22, 66.32, 57.62, 51.66, 43.92, 43.29, 41.30, 40.81, 32.93, 31.24, 30.38, 28.55, 19.21; ESI-MS m / z: 538.1894 calcd for C 29 H 32 NO7S + [M+H] + 538.1891.

[0106] Example 16

[0107] The compound 16 of the present application and the preparation method are shown as follows:

[0108]

[0109] Compound (VI) (30 mg, 0.06 mmol) was dissolved in 5 mL ethylene glycol dimethyl ether, NaOH (7 mg, 0.2 mmol) and methyl chloroacetate (0.08 mg, 0.07 mmol) were added, heated to reflux at 70 °C for 4 h, TLC detection of raw materials disappeared reaction was completed; a small amount of water was added to quench the reaction, extracted with dichloromethane three times, washed with saturated brine, dried and filtered, concentrated in vacuum, column chromatography to obtain white solid 15 mg (yield 47%). 1 H NMR (500 MHz, CDC13) δ 8.91 (t, J = 1.6 Hz, 1H), 8.84 (ddd, J = 3.8, 2.1, 1.4 Hz, 1H), 8.34 (dt, J = 8.2, 2.0 Hz, 1H), 7.59 (dd, J = 8.3, 4.0 Hz, 1H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.80 - 3.73 (m, 2H), 3.70 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.02 (s, 4H); 13 CNMR (125 MHz, CDC13) δ 207.74, 153.58, 149.71, 149.58, 142.75, 134.46, 131.28, 128.17, 124.91, 118.42, 101.75, 84.20, 68.94, 65.11, 63.61, 61.78, 59.10, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77; ESI-MS m / z: 514.1894 calcd for C 27 H 32 N6O2 [M+H] + 514.1903.

[0110] Example 17

[0111] The compound 17 of the present application and the preparation method are shown as follows:

[0112]

[0113] The synthesis method of compound 16 in Example 16 was referred to. 1 H NMR (500 MHz, CDC13) δ 7.50 (s, 1H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 4.03 (s, 3H), 3.80 - 3.73 (m, 2H), 3.70 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.3, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.02 (s, 5H); 13 CNMR (125 MHz, CDC13) δ 207.74, 153.58, 134.79, 131.28, 128.17, 124.30, 120.31, 118.42, 101.75, 85.55, 68.94, 65.11, 63.61, 61.78, 59.10, 51.79, 48.51, 47.08, 42.12, 40.99, 38.86, 32.99, 29.80, 29.75, 24.01, 19.77; ESI-MS m / z: 517.2003 calcd for C 26 H 37 N2O7S + [M+H] + 517.1997.

[0114] Example 18

[0115] The compound 18 of the present application and the preparation method are shown as follows:

[0116]

[0117] The synthesis method of compound 16 in Example 16 was referred to. 1H NMR (500 MHz, CDC13) δ 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 7.0, 3.8 Hz, 1H), 3.80 - 3.73 (m, 2H), 3.69 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 3.01 (t, J = 8.5 Hz, 2H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.94 (d, J = 8.6 Hz, 1H), 1.93 - 1.80 (m, 2H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.10 (t, J = 8.6 Hz, 3H), 1.02 (s, 5H); 13 C NMR (125 MHz, CDC13) δ 207.74, 153.58, 131.28, 128.17, 118.42, 101.75, 87.40, 68.94, 65.11, 63.61, 61.78, 59.10, 54.35, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77, 17.85, 13.09; ESI-MS m / z: 479.2098 calcd for C 25 H 35 O7S + [M+H] + 479.2090.

[0118] Example 19

[0119] The compound 19 of the present application and the preparation method are shown as follows:

[0120]

[0121] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 7.42 - 7.34 (m, 3H), 5.88 (dtt, J = 8.3, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 7.0, 3.8 Hz, 1H), 4.29 (t, J = 0.7 Hz, 2H), 3.80 - 3.73 (m, 2H), 3.69 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.84 (ddd, J = 15.2, 5.1, 0.9 Hz, 1H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.02 (s, 5H); 13 C NMR (125 MHz, CDC13) δ 207.74, 153.58, 131.54, 131.28, 131.09, 129.41, 128.19, 128.17, 118.42, 101.75, 87.40, 68.94, 65.11, 63.61, 61.78, 59.10, 58.00, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77; ESI-MS m / z: 527.2098 calcd for C 29 H 35 O7S + [M+H] + 527.2104.

[0122] Example 20

[0123] The compound 20 of the present application and the preparation method are shown as follows:

[0124]

[0125] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 9.06 (dd, J = 4.1, 1.7 Hz, 1H), 8.59 (d, J = 2.0 Hz, 1H), 8.26 (dd, J = 7.5, 1.6 Hz, 1H), 8.11 - 8.02 (m, 2H), 7.48 - 7.43 (m, 1H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.80 - 3.73 (m, 2H), 3.70 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.55 (m, 2H), 1.58 - 1.49 (m, 1H), 1.43 - 1.24 (m, 3H), 1.02 (s, 5H); 13 CNMR (125 MHz, CDC13) δ 207.74, 153.58, 150.81, 147.12, 137.42, 135.83, 132.01, 131.70, 131.28, 128.17, 127.22, 125.08, 121.20, 118.42, 101.75, 84.20, 68.94, 65.11, 63.61, 61.78, 59.10, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77; ESI-MS m / z: 564.2050 calcd for C 31 H 34 NO7S + [M+H] + 564.2093.

[0126] Example 21

[0127] The compound 21 of the present application and the preparation method are shown as follows:

[0128]

[0129] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 7.84 - 7.78 (m, 2H), 7.73 - 7.67 (m, 2H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.80 - 3.73 (m, 2H), 3.69 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.02 (s, 5H); 13 C NMR (125 MHz, CDC13) δ 207.74, 153.58, 140.08, 132.92, 131.28, 129.86, 129.84, 128.17, 118.42, 101.75, 84.20, 68.94, 65.11, 63.61, 61.78, 59.10, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77; ESI-MS m / z: 591.1047 calcd for C 28 H 32 BrO7S + [M+H] + 591.1045.

[0130] Example 22

[0131] The compound 22 of the present application and the preparation method are shown as follows:

[0132]

[0133] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 7.99 - 7.93 (m, 2H), 7.84 - 7.77 (m, 1H), 7.72 - 7.64 (m, 2H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.80 - 3.73 (m, 2H), 3.70 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.02 (s, 5H); 13 CNMR (125 MHz, CDC13) δ 207.74, 153.58, 138.83, 133.46, 131.28, 129.36, 128.17, 127.65, 118.42, 101.75, 84.20, 68.94, 65.11, 63.61, 61.78, 59.10, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77; ESI-MS m / z: 513.1942 calcd for C 28 H 33 O7S + [M+H] + 513.1947.

[0134] Example 23

[0135] The compound 23 of the present application and the preparation method are shown as follows:

[0136]

[0137] Refer to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.80 - 3.73 (m, 2H), 3.69 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.94 (p, J = 7.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 2.03 - 1.91 (m, 2H), 1.84 (ddd, J = 15.2, 5.1, 0.9 Hz, 1H), 1.78 - 1.24 (m, 11H), 1.02 (s, 4H); 13 C NMR (125 MHz, CDC13) δ 207.74, 153.58, 131.28, 128.17, 118.42, 101.75, 87.74, 68.94, 65.11, 63.61, 61.78, 60.93, 59.10, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 26.02, 24.42, 24.01, 23.52, 19.77; ESI-MS m / z: 519.2411 calcd for C 28 H 39 O7S + [M+H] + 519.2418.

[0138] Example 24

[0139] The compound 24 of the present application and the preparation method are shown as follows:

[0140]

[0141] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 8.06 - 8.00 (m, 2H), 7.95 - 7.89 (m, 2H), 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.85 (s, 2H), 3.80 - 3.73 (m, 2H), 3.69 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.68 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.02 (s, 4H); 13 C NMR (125 MHz, CDC13) δ 207.74, 167.06, 153.58, 143.39, 131.28, 131.26, 130.99, 130.74, 128.17, 118.42, 101.75, 84.20, 68.94, 65.11, 63.61, 61.78, 59.10, 52.18, 51.79, 48.51, 47.08, 42.12, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77; ESI-MS m / z: 571.1996 calcd for C 30 H 35 O9S + [M+H] + 571.2005.

[0142] Example 25

[0143] The compound 25 of the present application and the preparation method are shown as follows:

[0144]

[0145] Reference is made to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 5.88 (dtt, J = 8.2, 1.8, 0.9 Hz, 1H), 5.69 (d, J = 3.3 Hz, 2H), 5.36 (dt, J = 8.1, 4.9 Hz, 1H), 4.89 (dd, J = 6.9, 3.8 Hz, 1H), 3.80 - 3.73 (m, 2H), 3.70 - 3.62 (m, 2H), 3.54 (d, J = 7.7 Hz, 1H), 3.38 (dd, J = 10.2, 0.9 Hz, 1H), 2.83 - 2.73 (m, 2H), 2.42 (dddd, J = 12.8, 8.4, 5.9, 2.6 Hz, 1H), 1.88 - 1.80 (m, 1H), 1.72 (p, J = 7.8 Hz, 1H), 1.67 - 1.49 (m, 3H), 1.43 - 1.24 (m, 3H), 1.02 (s, 5H), 0.78 - 0.70 (m, 2H), 0.53 - 0.44 (m, 2H); 13 CNMR (125 MHz, CDC13) δ 207.74, 153.58, 131.28, 128.17, 118.42, 101.75, 87.74, 68.94, 65.11, 63.61, 61.78, 59.10, 51.79, 48.51, 47.08, 42.12, 41.79, 40.99, 32.99, 29.80, 29.75, 24.01, 19.77, 9.76; ESI-MS m / z: 477.1942 calcd for C 25 H 33 O7S + [M+H] + 477.1951.

[0146] Example 26

[0147] The compound 26 of the present application and the preparation method are shown as follows:

[0148]

[0149] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 7.86-7.80 (m, 2H), 7.45-7.38 (m, 2H), 5.74-5.65 (m, 2H), 5.49-5.40 (m, 2H), 5.04 (dd, J = 6.8, 3.8 Hz, 1H), 4.28 (d, J = 7.7 Hz, 1H), 4.12 (dd, J = 10.3, 0.9 Hz, 1H), 4.04 (ddd, J = 11.3, 4.9, 2.4 Hz, 1H), 3.97 (ddd, J = 11.7, 5.0, 2.3 Hz, 1H), 3.93-3.82 (m, 2H), 3.85-3.77 (m, 1H), 3.65-3.59 (m, 4H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.50 (tdd, J = 5.7, 2.5, 1.7 Hz, 1H), 2.05 (dddd, J = 13.5, 8.8, 5.8, 3.2 Hz, 1H), 2.01-1.81 (m, 5H), 1.78-1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 201.44, 172.64, 149.76, 139.03, 138.79, 135.12, 130.52, 127.90, 125.89, 118.23, 100.80, 82.92, 75.88, 66.39, 65.16, 63.06, 58.48, 51.90, 50.81, 43.86, 43.83, 41.23, 40.93, 40.62, 32.98, 31.21, 30.19, 28.54, 19.32; ESI-MS m / z: 585.2153 calcd for C 31 H 37 O9S + [M+H] + 585.2151.

[0150] Example 27

[0151] The compound 27 of the present application and the preparation method are shown as follows:

[0152]

[0153] Reference is made to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 7.44 (d, J = 3.1 Hz, 1H), 6.99 (dd, J = 5.5, 1.6 Hz, 1H), 6.82 (dd, J = 5.5, 1.3 Hz, 1H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.11 (dd, J = 7.0, 3.8 Hz, 1H), 4.28 (d, J = 7.7 Hz, 1H), 4.12 (dd, J = 10.3, 0.9 Hz, 1H), 4.04 (ddd, J = 11.3, 4.9, 2.4 Hz, 1H), 3.97 (ddd, J = 11.7, 5.0, 2.3 Hz, 1H), 3.88 (dddd, J = 21.1, 11.7, 5.0, 2.4 Hz, 2H), 3.80 (dd, J = 10.3, 0.8 Hz, 1H), 3.32 (dddd, J = 8.4, 6.6, 3.2, 2.4 Hz, 1H), 2.50 (tdd, J = 5.7, 2.5, 1.7 Hz, 1H), 2.05 (dddd, J = 13.6, 8.8, 5.8, 3.2 Hz, 1H), 2.01 - 1.81 (m, 5H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 201.96, 149.87, 146.94, 144.73, 139.03, 125.89, 118.23, 115.25, 113.56, 100.96, 81.91, 76.02, 66.39, 65.16, 63.06, 59.16, 50.81, 43.98, 43.79, 41.23, 40.93, 32.98, 31.24, 30.19, 28.54, 19.32; ESI-MS m / z: 503.1734 calcd for C 26 H 31 O8S + [M+H]+503.1736.

[0154] Example 28

[0155] The compound 28 of the present application and the preparation method are shown as follows:

[0156]

[0157] Reference is made to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 7.31 - 7.23 (m, 3H), 7.18 (ddddt, J = 5.0, 4.0, 3.1, 2.0, 1.0 Hz, 2H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.09 (dd, J = 7.1, 3.8 Hz, 1H), 4.28 (d, J = 7.7 Hz, 1H), 4.12 (dd, J = 10.2, 0.9 Hz, 1H), 4.04 (ddd, J = 11.3, 4.9, 2.4 Hz, 1H), 3.97 (ddd, J = 11.7, 5.0, 2.3 Hz, 1H), 3.93 - 3.82 (m, 2H), 3.85 - 3.77 (m, 1H), 3.42 - 3.23 (m, 3H), 3.01 (ddt, J = 11.1, 10.2, 0.9 Hz, 2H), 2.52 - 2.45 (m, 1H), 2.05 (dddd, J = 13.5, 8.8, 5.8, 3.2 Hz, 1H), 2.01 - 1.81 (m, 5H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 202.02, 149.76, 139.02, 135.37, 129.63, 128.70, 127.16, 125.89, 118.23, 101.27, 80.46, 76.05, 66.42, 65.16, 63.06, 58.34, 54.08, 50.81, 43.72, 43.59, 41.22, 40.93, 32.98, 31.22, 31.00, 30.19, 28.54, 19.35; ESI-MS m / z: 541.2255 calcd for C 30 H 37 O7S + [M+H] + 541.2259.

[0158] Example 29

[0159] The compound 29 of the present application and the preparation method are shown as follows:

[0160]

[0161] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 9.58 (s, 1H), 7.84-7.78 (m, 2H), 7.69-7.63 (m, 2H), 5.74-5.65 (m, 2H), 5.49-5.40 (m, 2H), 5.04 (dd, J = 6.8, 3.8 Hz, 1H), 4.28 (d, J = 7.7 Hz, 1H), 4.12 (dd, J = 10.2, 0.9 Hz, 1H), 4.04 (ddd, J = 11.3, 4.9, 2.4 Hz, 1H), 3.97 (ddd, J = 11.7, 5.0, 2.3 Hz, 1H), 3.88 (dddd, J = 21.1, 11.7, 5.0, 2.4 Hz, 2H), 3.80 (dd, J = 10.3, 0.8 Hz, 1H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.50 (tdd, J = 5.7, 2.5, 1.7 Hz, 1H), 2.05 (dddd, J = 13.6, 8.8, 5.8, 3.2 Hz, 1H), 2.01-1.81 (m, 5H), 1.73 (ddt, J = 12.9, 8.6, 5.5 Hz, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 201.44, 169.62, 149.76, 143.62, 139.03, 130.38, 129.43, 125.89, 120.89, 118.23, 100.80, 82.95, 75.88, 66.39, 65.16, 63.06, 58.48, 50.81, 43.86, 43.83, 41.23, 40.93, 32.98, 31.21, 30.19, 28.54, 24.01, 19.32; ESI-MS m / z: 570.2156 calcd for C 30 H 36 NO8S + [M+H] + 570.2158.

[0162] Example 30

[0163] The compound 30 of the present application and the preparation method are shown as follows:

[0164]

[0165] Reference to the synthesis method of compound 16 in Example 16. 1H NMR (500 MHz, CDC13) δ 8.77 (d, J = 6.8 Hz, 1H), 8.19 (t, J = 2.1 Hz, 1H), 7.74 (dd, J = 7.8, 2.1 Hz, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.22 - 7.17 (m, 1H), 6.64 (dd, J = 3.2, 2.2 Hz, 1H), 5.74 - 5.65 (m, 2H), 5.49 - 5.40 (m, 2H), 5.04 (dd, J = 6.8, 3.8 Hz, 1H), 4.28 (d, J = 7.7 Hz, 1H), 4.12 (dd, J = 10.3, 0.9 Hz, 1H), 4.04 (ddd, J = 11.3, 4.9, 2.4 Hz, 1H), 3.97 (ddd, J = 11.7, 5.0, 2.3 Hz, 1H), 3.93 - 3.77 (m, 3H), 3.32 (dddd, J = 6.8, 5.8, 4.5, 2.9 Hz, 1H), 2.50 (tdd, J = 5.7, 2.5, 1.7 Hz, 1H), 2.05 (dddd, J = 13.5, 8.8, 5.8, 3.2 Hz, 1H), 2.01 - 1.81 (m, 5H), 1.78 - 1.68 (m, 1H), 1.08 (s, 2H), 1.02 (s, 2H); 13 C NMR (125 MHz, CDC13) δ 201.44, 149.76, 139.03, 138.62, 130.85, 129.59, 125.89, 125.75, 123.58, 121.32, 118.23, 111.86, 103.13, 100.80, 82.95, 75.88, 66.39, 65.16, 63.06, 58.48, 50.81, 43.86, 43.83, 41.23, 40.93, 32.98, 31.21, 30.19, 28.54, 19.32; ESI-MS m / z: 552.2051 calcd for C 30 H 34 NO7S + [M+H] + 552.2050.

[0166] Example 31

[0167] Tablet formulation was prepared from compound 14 prepared in Example 14 according to the formulation in Table 1:

[0168] Table 1, tablet formulation

[0169]

[0170]

[0171] The tablet containing the ent-6,7-closed celangulin type oridonin hydroxyl modified derivative of the application is obtained.

[0172] Example 32

[0173] The ent-6,7-closed celangulin type oridonin hydroxyl modified derivative of the application is verified by pharmacological experiment:

[0174] Experimental equipment and reagents:

[0175] Ultra-clean workbench (Suzhou Aikelin Purification Equipment Co., Ltd.)

[0176] Constant temperature CO2 incubator (Japan SANYO)

[0177] Enzyme-linked immunoassay instrument (BIO-RAD, USA)

[0178] Inverted biological microscope (Japan OLYMPUS)

[0179] Reagent mixture of gentamicin (Nanjing Kayee Biotech Development Co., Ltd.)

[0180] Trypsin digestion solution (Nanjing Kayee Biotech Development Co., Ltd.)

[0181] PBS (Nanjing Kayee Biotech Development Co., Ltd.)

[0182] MTT (BIOSHARP)

[0183] DMSO (SIGMA)

[0184] Cell strain human breast cancer cell MCF-7

[0185] Human chronic myelogenous leukemia cell K562

[0186] Human colon cancer cell SW480

[0187] Human hepatocellular carcinoma cell SMMC-7721

[0188] Human breast cell MCF-10A

[0189] 1) Cell digestion, counting, making cell suspension with a concentration of 3×10 4 6 / mL, 100 uL of cell suspension was added to each well of 96-well plate (5×10 3 4 cells per well);

[0190] (2) The 96-well plate was placed in a 37℃, 5% CO2 incubator for 24 hours;

[0191] (3) Dilute the drug to the required concentration with complete culture medium, add 100 μL of the corresponding drug-containing culture medium to each well, and set up a negative control group, a solvent control group, and a positive control group.

[0192] (4) The 96-well plate was placed in a 37℃, 5% CO2 incubator for 72h; the 96-well plate was stained with MTT, λ=490nm, and the OD value was measured.

[0193] (5) Add 20 μL of MTT (5 mg / mL) to each well and continue incubation in an incubator for 4 hours;

[0194] (6) Discard the culture medium, add 150 μL of DMSO to each well to dissolve, and gently mix by shaking for 10 minutes; λ = 490 nm, read the OD value of each well with a microplate reader, and calculate the inhibition rate:

[0195] Cell inhibition rate % = 100% × (OD value of negative control group - OD value of compound group) / OD value of negative control group. The experimental results of the effect of the compound on the growth of human cancer cells and normal cells are shown in Table 2:

[0196] Table 2. Effects of some compounds on the growth of five human cell lines in vitro (IC50) 50 (Unit: μmol / ml)

[0197]

[0198] As shown in Table 2, oridonin derivatives inhibited all four types of cancer cells. Most derivatives exhibited stronger inhibitory activity than oridonin, and showed good activity against MCF-7 cells, with the greatest increase in inhibitory effect compared to oridonin, the best showing an activity more than 14 times that of oridonin. Furthermore, cyclization at positions 6 and 7 showed better activity than cyclization at positions 5; the introduction of oxygen and nitrogen heterocycles also improved activity and water solubility; and good safety was observed in normal breast cells MCF-10A.

[0199] The effects of the ent-6,7-closed-ring kaurene-type oridonin hydroxyl-modified derivative of the present invention on in vivo tumor growth were verified:

[0200] Female ICR mice (from Shanghai Sijie Experimental Animal Center) with a body weight of 18-22 g and successfully inoculated with breast cancer cells MCF-7 cells were selected and randomly divided into groups, 8 in each group. One group was a blank control, injected with the same volume of normal saline solution; another group was a positive drug control group, given 25 mg / kg of oridonin (purchased from Merck Company, Germany, with a goods number of 496919); the remaining groups were respectively given a certain dose of the compound. The solvents of oridonin and the above-mentioned compound were DMF: Tween 80: 5% normal saline = 10:2:88 (V:V:V), and the administration mode was 1 time per day, continuously for 21 days. After the experiment, the mice were killed and the tumors were surgically removed and weighed. The obtained data were statistically analyzed by SPSS 17.0:

[0201]

[0202] The results of the in-vivo inhibition of tumor growth of the compound are shown in Table 3:

[0203] Table 3, results of the in-vivo inhibition of tumor growth of the compound

[0204] Group Drug Dose Tumor inhibition rate 1 Saline - - 2 Compound 1 25 mg / kg 56.4％ 3 Compound 2 25 mg / kg 72.2％ 4 Compound 3 25 mg / kg 53.2％ 5 Compound 10 25 mg / kg 64.1％ 6 Compound 16 25 mg / kg 65.7％ 7 Compound 17 25 mg / kg 78.4％ 8 Compound 18 25 mg / kg 64.3％ 9 Compound 25 25 mg / kg 67.4％ 10 Oridonin 25 mg / kg 51.2％

[0205] As shown in Table 3, the oridonin derivatives have good in-vivo anti-tumor activity compared with oridonin, and are consistent with the in-vitro activity data.

[0206] The pharmacological test proves that the ent-6,7-cycloartane type oridonin hydroxyl modified derivative of the application has an effect of inhibiting the growth of cancer cells, and can be used for preparing an anti-tumor disease drug. Preferably, the treated tumor disease is breast cancer, leukemia, colon cancer and liver cancer; and the corresponding tumor cell strains used for the pharmacological test are MCF-7, K562, SW480 and SMC-7721 cell strains, which significantly inhibit the proliferation of breast cancer and liver cancer and other malignant tumor cells.

[0207] Example 33

[0208] Water-solubility experiment of the ent-6,7-cycloartane type oridonin hydroxyl modified derivative of the application:

[0209] 1. Experimental method

[0210] (1) Experimental instruments and conditions

[0211] HPLC model: Agilent 1100; chromatographic column: Lichrospher C18 (5 μm, 4.6*250 mm); column temperature: 37 ℃; flow rate: 1.0 ml / min; injection volume: 10 μL; detection wavelength: 210 nm.

[0212] (2) Experimental steps

[0213] Each sample was accurately weighed and prepared into standard solutions with concentrations of 0.01, 0.05, 0.1, 0.2, 0.4, 1.0 mg / mL using methanol, the main peak area was obtained by HPLC injection, and the standard curve was calculated. Then, an appropriate excess of the test compound (>10 mg) was dissolved in PBS, and incubated at 37°C on a constant temperature shaker for 24 hours. 10 μL of supernatant was taken by centrifugation and injected into HPLC to determine the main peak area of each compound, and the solubility was calculated by the standard curve. The experimental results are shown in Table 4.

[0214] 2. Experimental results

[0215] Table 4. Water solubility results of some compounds of the present application

[0216]

[0217] As can be seen from Table 4, compared with oridonin, the compounds designed in the present application have good water solubility, which is beneficial to drug development.

[0218] Example 34

[0219] Hepatic microsomal stability experiment of ent-6,7-closed ring kaurene type oridonin hydroxyl modified derivative of the present application:

[0220] The compound was co-incubated with human or mouse liver microsomes, and coenzyme NADPH was added to start the reaction. At incubation times of 0 min, 5 min, 15 min, 30 min, 60 min and 120 min, 20 μL of incubation solution was taken and transferred to 200 μL of acetonitrile containing an internal standard to terminate the reaction. After protein precipitation, centrifugation at 3700 rpm for 10 min, the supernatant was taken. The supernatant was diluted 1:1 with water and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS / MS) method. The detection results were processed using Graphpadprism 8.3.0, and the phase I metabolic stability parameters t 1 / 2 (min) of the compound in liver microsomes of different species were calculated.

[0221] Table 5. Hepatic microsomal stability of some compounds

[0222]

[0223] Therefore, the ent-6,7-closed ring kaurene type oridonin hydroxyl modified derivative of the present application can significantly inhibit the proliferation of various malignant tumor cells such as breast cancer and liver cancer, has extremely strong anti-tumor activity, has good pharmacokinetic stability and solubility, is beneficial to drug development, has low toxicity and side effects, and has great prospects in clinical application.

Claims

1. A hydroxyl-modified derivative of ent-6,7-closed-cyclic kaurene-type oridonin, characterized in that, The derivative is an oridonin derivative modified with an A-ring and hydroxyl groups at positions 6, 7, and 14, having the structure of Formula I, and also containing its pharmaceutically acceptable salt: ； The derivative is selected from any of the following structures: 。 2. An ent-6,7-closed-cyclic kaurene-type oridonin hydroxyl group A method for preparing modified derivatives, characterized by comprising the following steps: (1) Dissolve oridonin in anhydrous acetone, add 2,2-dimethoxypropane and p-toluenesulfonic acid, reflux the reaction, cool to room temperature after the reaction is completed, extract, wash, dry and concentrate to obtain compound II; (2) Compound II was dissolved in dichloromethane, triethylamine was added, and methanesulfonyl chloride was added dropwise to carry out the reaction. After the reaction was completed, the mixture was cooled to room temperature, extracted, washed, dried and concentrated to obtain compound III. (3) Dissolve compound III in DMF, add lithium carbonate and lithium bromide to react, cool to room temperature after the reaction is complete, filter, wash, concentrate, evaporate to dryness, and quickly pass dichloromethane through a column to obtain compound IV; (4) Compound IV was dissolved in tetrahydrofuran, and hydrochloric acid solution was added to react. After the reaction was completed, the solution was evaporated to dryness, washed with water, concentrated, and subjected to column chromatography to obtain compound V. (5) Compound V was dissolved in dichloromethane and reacted at room temperature. After the reaction was completed, a small amount of water was added to quench the reaction. The mixture was then extracted, washed, dried, filtered, concentrated under vacuum, and subjected to column chromatography to obtain compound VI. The reaction is carried out under reflux; or compound VI is dissolved in ethylene glycol dimethyl ether, and NaOH and methyl chloroacetate are added and the mixture is heated under reflux. After the reaction is completed, the mixture is concentrated under vacuum and column chromatography is performed to obtain ent-6,7-closed-ring kaurene-type oridonin hydroxyl-modified derivative I. The general reaction formula is as follows, where R is selected from C1-C6 alkyl, C1-C6 alkoxy, C 6-10 Aryl, C 3-8 Cycloalkyl, aziridine, thiocyclopropane, oxacyclobutane, pyridine, pyrimidine, furan, imidazole, pyrazole, quinoline, indole, benzofuran; or R is substituted by 1-4 identical or different R1s: R1 is selected from halogen, nitro, -NH2, C1-C6 alkoxy, C1-C6 alkylamino; n is 1 or 2; 。 3. A pharmaceutical composition, characterized in that, It comprises the derivative of claim 1, or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.

4. The ent-6,7-closed-cyclic kaurene-type oridonin hydroxyl group of claim 1 The use of the modified derivative or the pharmaceutical composition of claim 3 in the preparation of an antitumor drug, characterized in that, The tumor is a triple-negative breast cancer, leukemia, colon cancer, or liver cancer.

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