CN106243182B - Enoxolone-hydrogen sulfide donor reagent derivatives and its synthetic method and application - Google Patents
Enoxolone-hydrogen sulfide donor reagent derivatives and its synthetic method and application Download PDFInfo
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- CN106243182B CN106243182B CN201610621147.1A CN201610621147A CN106243182B CN 106243182 B CN106243182 B CN 106243182B CN 201610621147 A CN201610621147 A CN 201610621147A CN 106243182 B CN106243182 B CN 106243182B
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- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
Abstract
The invention discloses a kind of enoxolone-hydrogen sulfide donor reagent derivatives and its synthetic method and applications.The synthetic method of the derivative is：Extracting liquorice hypo acid, α, ω-two bromoalkane and alkali react in aprotic polar solvent, obtain compound 1；It takes compound 1, hydrogen sulfide donor reagent and alkali to react in aprotic polar solvent, obtains object crude product；Wherein, reaction carries out under conditions of being heated or not heated.Synthesis, which obtains the majority of compounds in derivative, has certain inhibitory activity to chronic myeloid leukemia cells K562, is expected to be used for corresponding anti-tumor drug and treats the preparation of the drug of chronic myelogenous leukemia.The obtained derivative that synthesizes has structure shown in the following general formula (I)：Wherein, n is 2~8；R isOr
Description
Technical field
The present invention relates to pharmaceutical technology fields, and in particular to a kind of enoxolone-hydrogen sulfide donor reagent derivatives and its
Synthetic method and application.
Background technique
Hydrogen sulfide is the new bioactivity gas molecule of one in succession after CO and NO, is the important angle for supporting life
Color has irreplaceable physiological regulatory action in vital movement, controls signal transduction process in various kinds of cell and plays product
The adjustment effect of pole.It is concentrated mainly on nerve, cardiovascular system about the potential treatment application of hydrogen sulfide at present, is such as treated high
Blood pressure, treatment heart ischemia disease, treat atherosclerosis, are combined with non-steroid anti-inflammatory drug, for reducing metabolism, prevent
Only conditions associated with hypoxia damage etc..
Hydrogen sulfide donor can hydrolyze in physiological conditions puts out H certainly2S discharges under cysteine (GSH) effect
H2The S sulphur atom that S, GSH can receive sulfide forms GSSH, is then urged by 3-mercaptopyruvate sulfurtransferase (3-MST)
Change and generates H2S。H2The biological effect of S and the research of signal path mechanism disclose it and are related to as signaling molecule in angiocarpy
The cell signalling of many organs such as system, nervous system, the circulatory system all has an impact to physiology courses multiple in body.
Enoxolone is the primary pharmacological activity substance of Radix Glycyrrhizae.In recent years, not with the research of enoxolone pharmacological action
Disconnected deeply pharmacological action is increasingly realized, including antitumor, anti-inflammatory, antiviral, treatment cardiovascular disease, immunological regulation, anti-
The multiple pharmacological effects such as oxidation.Using natural products as lead compound, structural modification is carried out to it and introduces corresponding active drug effect
Group, and then carry out the pharmacology activity research in corresponding field, it has also become the research hotspot of new drug development.
It has not yet to see enoxolone and derivative and its synthesis side of the hydrogen sulfide donor reagent through alkane chain link
The relevant report of method and application.
Summary of the invention
It is derivative that the technical problem to be solved in the present invention is to provide the enoxolone of a kind of structure novel-hydrogen sulfide donor reagents
Object and their synthetic method and application.
The present invention relates to the enoxolone with structure shown in the following general formula (I)-hydrogen sulfide donor reagent derivatives or its
Pharmaceutically acceptable salt：
Wherein,
N is 2~8；
R is
Enoxolone of the present invention-hydrogen sulfide donor reagent derivatives synthetic method is：Extracting liquorice hypo acid, α, ω-
Two bromoalkanes and alkali react in aprotic polar solvent, obtain compound 1；Compound 1, hydrogen sulfide donor reagent and alkali is taken to exist
It is reacted in aprotic polar solvent, obtains object crude product；Wherein, reaction carries out under conditions of being heated or not heated.
More specifically synthetic method includes the following steps：
1) extracting liquorice hypo acid, α, ω-two bromoalkane and alkali react in aprotic polar solvent, and gained reactant removes molten
Agent, residue is scattered in ethyl acetate, methylene chloride or ether, washed, after anhydrous sodium sulfate dries, filters, collects filter
Liquid obtains compound 1 after filtrate concentration；
2) compound 1, hydrogen sulfide donor reagent and alkali is taken to react in aprotic polar solvent, gained reactant removes molten
Agent, residue is scattered in ethyl acetate, methylene chloride or ether, washed, after anhydrous sodium sulfate dries, filters, collects filter
Liquid obtains object crude product after filtrate concentration.
In the step 1) and step 2) of above-mentioned specific synthetic method, the washing preferably successively uses hydrochloric acid, water, satisfies
It is washed with saline solution, or is successively washed with hydrochloric acid, saturated salt solution.
The structural formula of the compound 1 synthesized in synthetic method of the present invention is as follows：
Wherein, n is 2~8.
The compound 1 synthesized in the above method is the crude product of compound 1, in order to improve the purity of compound 1 simultaneously
It reduces in subsequent reactions and generates more by-products, preferably by the crude product of gained compound 1 through silica gel thin-layer chromatography or silica gel
Subsequent operation is used further to after column chromatographic purifying.When being carried out silica gel thin-layer chromatography or upper silica gel column chromatography, usually with by body
Product is than being 2~10：The eluent of 1 petroleum ether (PE) and ethyl acetate (EA) composition collects eluent, eluent decompression
Solvent is evaporated off, obtains object after purification.The petroleum ether of the composition eluant, eluent and the volume ratio of ethyl acetate be preferably 2~
5：1.
Prepared by the above method is the crude product of formula (I) compound, and existing conventional purification process can be used and carry out to it
It purifies to improve the purity of formula (I) compound.Silica gel thin-layer chromatography or silica gel column chromatography are generallyd use to be purified, will made
It is usually 2~10 with by volume ratio when the target compound crude product silica gel thin-layer chromatography or upper silica gel column chromatography that obtain：1 petroleum
The eluent of ether (PE) and ethyl acetate (EA) composition, collects eluent, and eluent evaporating solvent under reduced pressure obtains after purification
Object.The petroleum ether of the composition eluant, eluent and the volume ratio of ethyl acetate are preferably 2~5：1.
In synthetic method of the present invention, the α, ω-two bromoalkane can be 1,2- Bromofume, 1,3- dibromo
Propane, 1,4- dibromobutane, pentamethylene bromide, bis- bromooctane of 1,6- dibromo-hexane, 1,7- dibromo-heptane or 1,8-.
In synthetic method of the present invention, the alkali can be potassium carbonate, triethylamine, sodium carbonate, sodium bicarbonate, carbon
Potassium hydrogen phthalate or cesium carbonate.When alkali is when being selected as cesium carbonate, higher yield can be obtained；Comprehensively consider from cost and yield,
It is preferred that alkali is potassium carbonate.
In synthetic method of the present invention, the aprotic polar solvent can be n,N-Dimethylformamide
(DMF), the combination of one or more of toluene and pyridine, when the selection of aprotic polar solvent is more than above two
Combination when, the proportion between them can be any proportion.The dosage of the aprotic polar solvent is usually that can dissolve
Participate in the raw material of reaction.
In synthetic method of the present invention, the hydrogen sulfide donor reagent specifically can be p-hydroxybenzene -1 5-,
2- dithiole -3- thioketones (ADT-OH), (R)-lipoic acid (R-lipoic acid) or 4- hydroxythiobenzamide
(TBZ), their structural formula difference is as follows：
In synthetic method of the present invention, the enoxolone, α, the reaction of ω-two bromoalkane and alkali is preferably low
In or equal to carrying out under conditions of 40 DEG C, applicant has found in an experiment, can be with when reaction carries out under the conditions of 20~40 DEG C
Higher yield is obtained in a relatively short period of time, while side reaction is less；The compound 1, hydrogen sulfide donor reagent and alkali
Reaction carries out under conditions of being less than or equal to 65 DEG C, more preferably carries out under the conditions of 35~65 DEG C, in this way can be shorter
Time in obtain higher yield, and reduce the generation of by-product to the greatest extent.Under the conditions of above-mentioned limiting temperature, whether complete react
Thin-layer chromatography tracing detection can be passed through entirely.
In synthetic method of the present invention, the enoxolone, α, the ratio between ω-two bromoalkane and the amount of substance of alkali
For：1：1~5：0.5~3；The ratio between the compound 1, hydrogen sulfide donor reagent and amount of substance of alkali are：1：1~3：1~5.
It has been found that being added catalyst potassium iodide (KI) in the reaction of compound 1, hydrogen sulfide donor reagent and alkali
It can be further improved the yield of object.The additional amount of the potassium iodide is 0.1~1 times of the amount of 1 substance of compound.
Compared with prior art, the present invention provides a series of enoxolone of structure novels-hydrogen sulfide donor reagents to spread out
Biology and its synthetic method, meanwhile, it is white to tumor cells of hepatocellular carcinoma strain and chronic myelognous that applicant has also investigated these derivatives
The inhibitory activity of blood disease cell, the results showed that, wherein most compound has one to chronic myeloid leukemia cells K562
Fixed inhibitory activity is expected to be used for corresponding anti-tumor drug and treats the preparation of the drug of chronic myelogenous leukemia.
Specific embodiment
The present invention is described in further detail combined with specific embodiments below, content to better understand the invention, but
The present invention is not limited to following embodiments.
Enoxolone-hydrogen sulfide donor reagent derivatives of the present invention with structure shown in the following general formula (I) are pressed
Following synthetic routes are synthesized：
Wherein：
α, ω-two bromoalkane can be glycol dibromide, 1,3- dibromopropane, 1,4- dibromobutane, 1,5- dibromo penta
Alkane, bis- bromooctane of 1,6- dibromo-hexane, 1,7- dibromo-heptane or 1,8-；
Aprotic polar solvent can be N,N-dimethylformamide, toluene or pyridine；
Alkali can be potassium carbonate, triethylamine, sodium carbonate, sodium bicarbonate, saleratus or cesium carbonate；
Hydrogen sulfide donor reagent specifically can be 5- p-hydroxybenzene -1,2- dithiole -3- thioketones (ADT-OH),
(R)-lipoic acid (R-lipoic acid) or 4- hydroxythiobenzamide (TBZ), their structural formula difference are as follows：
N in compound 1 and compound 2 is 2~8；
R in compound 2 is
Embodiment 1：The synthesis of compound 1a
Enoxolone (500mg, 1.06mmol) is dissolved in anhydrous DMF (5mL), addition 1,6- Bromofume (2.43mL,
5.3mmol)、K2CO3(146.28mg, 1.06mmol), 30 DEG C of reactions are for 24 hours.Evaporating solvent under reduced pressure, residue disperse ethyl acetate
In (50mL), HCl (1N), water, saturated common salt water washing are successively used, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column
Chromatography (VPE:VEA=2:1) compound 1a (457mg, 75%, white solid), is obtained.
Yield:457mg, 75%, white solid；Rf=0.461 (Petroluem ether:EtOAc=2:1)
.M.p 190-192℃.1H NMR(500MHz,CDCl3)δ(ppm):5.69 (s, 1H, 12-H), 4.41 (dd, J=28.0,
5.9Hz,2H,OCH2), 3.53 (t, J=5.7Hz, 2H, OCH2), 3.21 (dd, J=11.0,5.2Hz, 1H, 3-H), 2.83-
2.71(m,1H,18-H),2.32(s,1H),2.21-0.63(m,20H),1.35,1.17,1.12,1.11,0.99,0.80and
0.79(7s,each 3H,7×CH3).13C NMR(125MHz,CDCl3)δ(ppm):200.3,176.2,169.2,128.8,
78.9,63.9,61.9,55.1,48.3,45.5,44.3,43.3,41.0,39.2,37.8,37.2,32.9,32.0,31.2,
29.2,28.6,28.2,27.4,26.6,23.5,18.8,17.6,16.5,15.7.HRMS(ESI)m/z:[M+H]+calcd
for C32H50BrO4,577.2893；found 577.2873.
Embodiment 2：The synthesis of compound 1b
Enoxolone (500mg, 1.06mmol) is dissolved in anhydrous DMF (5mL), addition 1,8- dibromobutane (2.94mL,
5.3mmol)、K2CO3(146.28mg, 1.06mmol), 30 DEG C of reactions are for 24 hours.Evaporating solvent under reduced pressure, residue disperse ethyl acetate
In (50mL), HCl (1N), water, saturated common salt water washing are successively used, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column
Chromatography (VPE:VEA=2:1) compound 1b (532mg, 83%, white solid), is obtained.
Yield:532mg, 83%, white solid；Rf=0.515 (Petroluem ether:EtOAc=2:1)
.M.p 82-84℃.1H NMR(500MHz,CDCl3)δ(ppm):5.61 (s, 1H, 12-H), 4.12 (t, J=6.3Hz, 2H,
OCH2), 3.43 (t, J=6.5Hz, 2H, OCH2), 3.21 (dd, J=11.1,5.1Hz, 1H, 3-H), 2.77 (d, J=13.5Hz,
1H,18-H),2.32(s,1H),2.13-0.61(m,24H),1.35,1.14,1.11,1.10and 0.98(5s,each 3H,5
×CH3),0.79(s,6H,2×CH3).13C NMR(125MHz,CDCl3)δ(ppm):200.3,176.5 169.3,128.6,
78.8,63.5,61.9,55.0,48.5,45.5,44.1,43.3,41.1,39.2,37.8,37.2,33.1,32.8,31.9,
31.2,29.4,28.6,28.2,27.4,26.5,23.5,18.8,17.6,16.5,15.7.HRMS(ESI)m/z:[M+H]+
calcd for C34H54BrO4,605.3206；found 605.3188.
Embodiment 3：The synthesis of compound 1c
Enoxolone (1.0g, 2.12mmol) is dissolved in anhydrous DMF (5mL), addition 1,6- dibromo-hexane (1.62mL,
10.62mmol)、K2CO3(293.0mg, 2.12mmol), 30 DEG C of reactions are for 24 hours.Evaporating solvent under reduced pressure, residue disperse ethyl acetate
In (50ml), HCl (1N), water, saturated common salt water washing are successively used, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column
Chromatography (VPE:VEA=5:2) compound 1c (929mg, 69%, white solid), is obtained.
Yield:929mg, 69%, white solid；Rf=0.452 (Petroluem ether:EtOAc=5:2)
.M.p 102-104℃.1H NMR(500MHz,CDCl3)δ(ppm):5.61(s,1H,12-H),4.08(m,2H,OCH2),3.39
(m,2H,CH2- Br), 3.20 (dd, J=11.1,5.2Hz, 1H, 3-H), 2.76 (d, J=13.5Hz, 1H, 18-H), 2.32 (s,
1H,10-H),2.09-0.69(m,35H),1.35,1.13,1.11,1.10and 0.98(5s,each 3H,5×CH3),0.79
(s,6H,2×CH3).13C NMR(125MHz,CDCl3)δ(ppm):200.3,176.6,169.4,128.6,61.9,78.8,
64.4,55.0,48.5,45.5,44.1,43.3,41.2,39.2,37.9,37.2,33.9,32.9,32.7 31.9,31.2,
28.7,28.5,28.2,27.8,27.4,26.6,26.5,25.4,23.5,18.8,17.6,16.5,15.7.HRMS(ESI)m/
z:[M+H]+calcd for C36H58BrO4,633.3519；found633.3525.
Embodiment 4：The synthesis of compound 1d
Enoxolone (1.0g, 2.12mmol) is dissolved in anhydrous DMF (5mL), addition 1,8-, bis- bromooctane (1.62mL,
10.62mmol)、K2CO3(293.0mg, 2.12mmol), 30 DEG C of reactions are for 24 hours.Evaporating solvent under reduced pressure, residue disperse ethyl acetate
In (50mL), HCl (1N), water, saturated common salt water washing are successively used, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column
Chromatography (VPE:VEA=5:2) compound 1d (984mg, 62%, white solid), is obtained.
Yield:984mg, 62%, white solid；Rf=0.500 (Petroluem ether:EtOAc=5:2)
.M.p 67-69℃.1H NMR(500MHz,CDCl3)δ(ppm):5.62(s,1H,12-H),4.07(m,2H,OCH2),3.39
(m,2H,CH2- Br), 3.21 (dd, J=11.1,5.2Hz, 1H, 3-H), 2.77 (d, J=13.6Hz, 1H, 18-H), 2.32 (s,
1H,10-H),2.10-0.69(m,35H),1.35,1.13,1.12,1.11and 0.99(5s,each 3H,5×CH3),0.79
(s,6H,2×CH3).13C NMR(125MHz,CDCl3)δ(ppm):200.3,176.6,169.4,128.6,78.9,64.6,
61.9,55.1,48.5,45.5,44.1,43.3,41.2,39.2,37.9,34.1,37.2,32.9,31.9,31.3,29.1,
28.7,28.8,28.7,28.6,28.3,28.2,27.4,26.6,26.5,26.0,23.5,18.8,17.6,16.5,
15.7.HRMS(ESI)m/z:[M+H]+calcd for C38H62BrO4,661.3831；found 661.3836.
Embodiment 5：The synthesis of compound 2a
Compound 1a (250mg, 0.43mmol) is dissolved in DMF (5mL), addition (R)-lipoicacid (88.58mg,
0.43mmol)、K2CO3(178.02mg, 1.29mmol), 50 DEG C of reactions are for 24 hours.Residue is dispersed in ethyl acetate (50mL), successively
With HCl (1N), water, saturated common salt water washing, anhydrous sodium sulfate is dried, filtered, filtrate decompression concentration, column chromatography for separation (VPE:
VEA=2:1) compound 2a (231mg, 76%, faint yellow solid), is obtained.
Yield:231mg, 76%, yellow solid；Rf=0.490 (Petroluem ether:EtOAc=2:1)
.M.p 64-66℃.1H NMR(500MHz,CDCl3)δ(ppm):5.62(s,1H,12-H),4.41-4.18(m,4H,2×
OCH2), 3.52 (dd, J=8.2,6.4Hz, 1H, 3-H), 3.25-2.98 (m, 3H), 2.75 (d, J=13.5Hz, 1H, 18-H),
2.44 (s, 1H), 2.33 (dd, J=14.7,7.2Hz, 3H), 2.14-0.60 (m, 26H), 1.34,1.13,1.10,1.09,
0.97,0.78and 0.77(7s,each 3H,7×CH3).13C NMR(125MHz,CDCl3)δ(ppm):200.1,176.2,
173.3,169.2,128.5,78.8,62.3,61.9,56.4,55.0,48.4,45.5,44.1,43.3,41.1,40.3,
39.2,38.6,37.8,37.2,34.6,34.0,32.8,31.9,31.2,28.8,28.7,28.3,28.2,27.3,26.5,
24.6,23.5,18.8,17.6,16.4,15.7,14.3.HRMS(ESI)m/z:[M+H]+calcdfor C40H62ClO6S2,
737.3676；found 737.3696.
Embodiment 6：The synthesis of compound 2b
Compound 1b (250mg, 0.41mmol) is dissolved in DMF (5mL) F, be added (R)-lipoicacid (85.12mg,
0.41mmol)、K2CO3(169.74mg, 1.23mmol), 50 DEG C of reactions are for 24 hours.Residue is dispersed in ethyl acetate (50mL), successively
With HCl (1N), water, saturated common salt water washing, anhydrous sodium sulfate is dried, filtered, filtrate decompression concentration, column chromatography for separation (VPE:
VEA=2:1) compound 2b (190mg, 63%, faint yellow solid), is obtained.
Yield:190mg, 63%, yellow solid；Rf=0.431 (Petroluem ether:EtOAc=2:1)
.M.p 60-62℃.1H NMR(500MHz,CDCl3)δ(ppm):5.60(s,1H,12-H),4.09(m,4H,2×OCH2),
3.54 (dd, J=8.2,6.4Hz, 1H, 3-H), 3.21-3.02 (m, 3H), 2.75 (dd, J=13.3,3.3Hz, 1H, 18-H),
2.45 (s, 1H), 2.30 (dd, J=9.8,4.7Hz, 3H), 2.15-0.61 (m, 30H), 1.34,1.13,1.10,1.09and
0.98(5s,each 3H,5×CH3),0.78(s,6H,2×CH3).13C NMR(125MHz,CDCl3)δ(ppm):200.2,
176.5,173.6,169.3,128.6,78.8,63.9,61.9,56.4,55.0,48.5,45.5,44.1,43.3,41.1,
40.3,39.2,38.6,37.8,37.2,34.7,34.1,32.8,31.9,31.2,28.8,28.7,28.5,28.2,27.4,
26.6,26.5,25.6,25.5,24.7,23.5,18.8,17.6,16.5,15.7.HRMS(ESI)m/z:[M+H]+calcd
for C42H66ClO6S2,765.3989；found 765.4008.
Embodiment 7：The synthesis of compound 2c
Compound 1c (500mg, 0.79mmol) is dissolved in DMF (5mL), addition (R)-lipoicacid (163.0mg,
0.79mmol)、K2CO3(327.58mg, 2.37mmol), 50 DEG C of reactions are for 24 hours.Residue is dispersed in ethyl acetate (50mL), successively
With HCl (1N), water, saturated common salt water washing, anhydrous sodium sulfate is dried, filtered, filtrate decompression concentration, column chromatography for separation (VPE:
VEA=3:1) compound 2c (402mg, 67%, faint yellow solid), is obtained.
Yield:402mg, 67%, yellow solid；Rf=0.339 (Petroluem ether:EtOAc=3:1)
.M.p 51-53℃.1H NMR(400MHz,CDCl3)δ(ppm):5.60(s,1H,12-H),4.05(m,4H,2×OCH2),
3.54 (dd, J=8.0,6.5Hz, 1H, 3-H), 3.25-3.03 (m, 3H), 2.75 (dd, J=10.1,3.4Hz, 1H, 18-H),
2.42(m,1H),2.29(m,2H),2.12-0.58(m,36H),1.35(s,3H,CH3),1.12–1.04(m,9H,3×CH3),
0.97(s,3H,CH3),0.77(s,6H,2×CH3).13C NMR(100MHz,CDCl3)δ(ppm):200.2,176.5,173.6,
169.3,128.6,78.8,64.4,61.9,56.4,55.0,48.5,45.5,44.1,43.3,41.2,40.3,39.2,38.5,
37.8,37.2,34.7,34.2,32.8,31.9,31.2,28.6,28.2,27.4,26.5,25.8,25.6,24.8,23.5,
18.8,17.6,16.4,15.7.HRMS(ESI)m/z:[M+H]+calcd for C44H71O6S2,759.4692；found
759.4696.
Embodiment 8：The synthesis of compound 2d
Compound 1d (500mg, 0.76mmol) is dissolved in DMF (5mL), addition (R)-lipoicacid (156.81mg,
0.76mmol)、K2CO3(315.12mg, 2.28mmol), 50 DEG C of reactions are for 24 hours.Residue is dispersed in ethyl acetate (50mL), successively
With HCl (1N), water, saturated common salt water washing, anhydrous sodium sulfate is dried, filtered, filtrate decompression concentration, column chromatography for separation (VPE:
VEA=3:1) compound 2d (475mg, 80%, faint yellow solid), is obtained.
Yield:475mg, 80%, yellow solid；Rf=0.578 (Petroluem ether:EtOAc=3:1)
.M.p 52-54℃.1H NMR(400MHz,CDCl3)δ(ppm):5.60(s,1H,H-12),4.13-3.97(s,4H,2×
OCH2), 3.53 (m, 1H, 3-H), 3.27 (m, 3H), 2.74 (d, J=13.5Hz, 1H, 18-H), 2.42 (dd, J=12.5,
6.2Hz, 1H), 2.28 (dd, J=13.7,6.3Hz, 3H), 2.10-0.60 (m, 40H), 1.12-1.05 (m, 9H, 2 × CH3),
0.96(s,6H,2×CH3),0.77(s,6H,2×CH3).13C NMR(100MHz,CDCl3)δ(ppm):200.2,176.5,
173.6,169.3,128.5,78.7,64.5,61.9,56.4,55.0,48.4,45.4,44.0,43.3,41.1,40.2,
39.2,38.5,37.8,37.1,34.6,34.1,32.8,31.9,31.2,29.1,28.8,28.7,28.6,28.5,28.2,
27.4,26.5,25.9,24.8,23.5,18.7,17.5,16.4,15.7.HRMS(ESI)m/z:[M+H]+calcd for
C46H75O6S2,787.5005；found 787.5019.
Embodiment 9：The synthesis of compound 2e
Compound 1c (500mg, 0.79mmol) is dissolved in DMF (5mL), addition ADT-OH (178.57mg, 0.79mmol),
K2CO3(327.58mg, 2.37mmol), KI (13.28mg, 0.08mmol), 65 DEG C of reactions are for 24 hours.Residue is dispersed in ethyl acetate
(50mL) successively uses HCl (1N), water, saturated common salt water washing, and anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column layer
Analysis separation (VPE:VEA=3:1) compound 2e (126mg, 21%, orange solids), is obtained.
Yield:126mg, 21%, orange solid；Rf=0.352 (Petroluem ether:EtOAc=3:1)
.M.p 92-94℃.1H NMR(400MHz,CDCl3)δ(ppm):7.60 (d, J=8.8Hz, 2H, Ar-H), 7.39 (s, 1H, Ar-
), H 6.96 (d, J=8.8Hz, 2H, Ar-H), 5.63 (s, 1H, 12-H), 4.22-3.97 (m, 4H, 2 × OCH2),3.23(dd,J
=10.9,5.3Hz, 1H, 3-H), 2.79 (d, J=13.5Hz, 1H, 18-H), 2.34 (s, 1H, 9-H), 2.13-0.69 (m,
29H),1.19-1.09(m,9H,3×CH3),1.01(s,6H,2×CH3),0.81(s,6H,2×CH3).13CNMR(100MHz,
CDCl3)δ(ppm):215.1,200.3,176.6,173.3,169.5,162.6,134.6,128.7,128.6,124.0,
115.6,78.8,68.3,64.4,61.9,55.0,48.6,45.5,44.1,43.3,41.2,39.2,37.8,37.2,32.9,
31.9,31.2,29.0,28.8,28.7,28.5,28.2,27.4,26.54,26.51,25.9,25.7,23.5,18.8,17.6,
16.5,15.7.HRMS(APCl)m/z:[M+H]+calcd for C45H63O5S3,779.3838；found 779.3820.
Embodiment 10：The synthesis of compound 2f
Compound 1d (500mg, 0.76mmol) is dissolved in DMF (5mL), addition ADT-OH (169.74mg, 0.76mmol),
K2CO3(315.12mg, 2.28mmol), KI (13.28mg, 0.08mmol), 65 DEG C of reactions are for 24 hours.Residue is dispersed in ethyl acetate
(50mL) successively uses HCl (1N), water, saturated common salt water washing, and anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column layer
Analysis separation (VPE:VEA=3:1) compound 2f (118mg, 19%, orange solids), is obtained.
Yield:118mg, 19%, orange solid；Rf=0.369 (Petroluem ether:EtOAc=3:1)
.M.p 87-89℃.1H NMR(400MHz,CDCl3)δ(ppm):7.60 (d, J=8.8Hz, 2H, Ar-H), 7.39 (s, 1H),
6.96 (d, J=8.8Hz, 2H, Ar-H), 5.64 (s, 1H, 12-H), 4.14-3.98 (m, 4H, 2 × OCH2), 3.22 (dd, J=
10.7,5.3Hz, 1H, 3-H), 2.78 (d, J=13.5Hz, 1H, 18-H), 2.34 (s, 1H, 9-H), 2.17-0.64 (m, 33H),
1.16-1.10(m,9H,3×CH3),1.00(s,3H,CH3),0.80(s,6H,2×CH3).13C NMR(100MHz,CDCl3)δ
(ppm):215.2,200.3,176.6,173.3,169.4,162.7,134.6,128.7,128.6,124.0,115.6,78.8,
68.5,64.6,61.9,55.1,48.5,45.5,44.1,43.3,41.2,39.2,37.9,37.2,32.9,31.9,31.3,
29.3,29.2,29.1,28.7,28.5,28.2,27.4,26.6,26.5,26.0,26.0,23.5,18.8,17.6,16.5,
15.7.HRMS(APCl)m/z:[M+H]+calcd forC47H67O5S3,807.4151；found 807.4118.
Embodiment 11：The synthesis of compound 2g
Compound 1c (250mg, 0.39mmol) is dissolved in DMF (5mL), TBZ (60.35mg, 0.39mmol), K is added2CO3
(28mg, 0.2mmol), KI (6.30mg, 0.04mmol), 35 DEG C of reaction 12h.Residue is dispersed in ethyl acetate (50mL), according to
Secondary to use HCl (1N), water, saturated common salt water washing, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column chromatography for separation (VPE:
VEA=3:1) compound 2g (91mg, 32%, light yellow solid), is obtained.
Yield:91mg, 32%, yellow solid；Rf=0.267 (Petroluem ether:EtOAc=3:1)
.M.p 119-120℃.1H NMR(500MHz,DMSO-d6)δ(ppm):9.30 (s, 1H, NH), 8.23 (s, 1H, NH), 7.48
(d, J=8.8Hz, 2H, Ar-H), 6.99 (d, J=8.9Hz, 2H, Ar-H), 5.40 (s, 1H, 12-H), 4.02-3.96 (m, 4H,
2×OCH2), 2.98 (d, J=4.5Hz, 1H, 3-H), 2.55 (d, J=13.2Hz, 1H, 18-H), 2.30 (s, 1H), 2.18-
0.63(m,28H),1.08,0.89and 0.72(3s,each 3H,3×CH3,1.00and 0.67(2s,each 6H,4×
CH3).13C NMR(125MHz,DMSO-d6)δ(ppm):199.1,191.7,175.9,169.5,161.6,134.2,126.4,
120.9,115.0,76.6,67.8,63.9,61.2,54.1,48.1,44.9,43.6,42.9,37.4,36.7,32.1,31.6,
30.4,29.1,28.7,28.6,28.5,28.32,28.28,28.2,27.8,27.0,26.1,25.8,25.6,25.4,23.0,
22.2,18.4,17.2,16.2,16.1.HRMS(ESI)m/z:[M+Na]+calcd for C43H64NO5S,706.4505；
found 706.4500.
Embodiment 12：The synthesis of compound 2h
Compound 1d (250mg, 0.38mmol) is dissolved in DMF (5mL), TBZ (57.80mg, 0.38mmol), K is added2CO3
(28mg, 0.20mmol), KI (6.30mg, 0.04mmol), 35 DEG C of reaction 12h.Residue is dispersed in ethyl acetate (50mL), according to
Secondary to use HCl (1N), water, saturated common salt water washing, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, column chromatography for separation (VPE:
VEA=3:1) compound 2h (107mg, 39%, light yellow solid), is obtained.
Yield:107mg, 39%, yellow solid；Rf=0.206 (Petroluem ether:EtOAc=3:1)
.M.p 117-119℃.1H NMR(500MHz,DMSO-d6)δ(ppm):9.63(s,1H,NH),9.31(s,1H,NH),7.94
(d, J=8.9Hz, 2H, Ar-H), 6.91 (d, J=8.9Hz, 2H, Ar-H), 5.42 (s, 1H, 12-H), 4.01 (m, 4H, 2 ×
OCH2), 3.01 (dd, J=11.4,4.4Hz, 1H, 3-H), 2.58 (s, 1H, 18-H), 2.31 (s, 1H), 2.15-0.64 (m,
32H),1.09,0.90and 0.73(3s,each 3H,3×CH3),1.02and 0.68(2s,each 6H,4×CH3).13C
NMR(125MHz,DMSO-d6)δ(ppm):199.0,198.5,175.8,169.4,161.4,131.1,129.5,127.4,
113.4,76.6,67.7,63.9,61.2,59.8,54.1,48.1,44.9,43.6,42.9,37.4,36.7,32.1,31.5,
30.4,28.5,28.2,27.8,27.0,26.1,25.8,25.4,25.2,23.0,20.8,18.4,16.2,16.0,
14.4.HRMS(ESI)m/z:[M+Na]+calcd forC45H68NO5S,734.4818；found 734.4816.
Embodiment 13：The synthesis of compound 2a
Embodiment 5 is repeated, unlike：DMF is substituted with toluene, substitutes K with cesium carbonate2CO3, and temperature is changed to 40 DEG C
React 12h.
Products therefrom (228mg, 75%, faint yellow solid) is accredited as compound 2a, the following institute of structural formula by nuclear-magnetism
Show：
Embodiment 14：The synthesis of compound 2b
Embodiment 6 is repeated, unlike：DMF is substituted with pyridine, substitutes K with saleratus2CO3, and being equivalent to is added
Close 0.5 times of amount of catalyst KI of object 1b substance.
Products therefrom (166mg, 55%, faint yellow solid) is accredited as compound 2b, the following institute of structural formula by nuclear-magnetism
Show：
Embodiment 15：The synthesis of compound 2e
Embodiment 9 is repeated, unlike：DMF is substituted with toluene, substitutes K with sodium carbonate2CO3, and it is added without catalyst KI,
It disperses residue in methylene chloride.
Products therefrom (72mg, 12%, orange solids) is accredited as compound 2e by nuclear-magnetism, and structural formula is as follows：
Embodiment 16：The synthesis of compound 2f
Embodiment 10 is repeated, unlike：With with the composition of toluene and DMF (1 is pressed by toluene and DMF：1 volume ratio
Composition) substitution DMF, K is substituted with triethylamine2CO3, and it is added without catalyst KI, it disperses residue in ether.
Products therefrom (62mg, 10%, orange solids) is accredited as compound 2f by nuclear-magnetism, and structural formula is as follows：
Embodiment 16：The synthesis of compound 2h
Embodiment 12 is repeated, unlike：With the composition of pyridine and toluene (1 is pressed by picolin and toluene：5 volume
Than composition) substitution DMF, K is substituted with sodium bicarbonate2CO3, and it is added without catalyst KI.
Products therefrom (49mg, 18%, light yellow solid) is accredited as compound 2h, the following institute of structural formula by nuclear-magnetism
Show：
Applicant is thin to human liver cancer tumor cell line and chronic myelogenous leukemia to compound 2a~2h of the present invention
The proliferation inhibition activity of born of the same parents is tested：
1, cell strain and cell culture
Human liver cancer cell BEL-7402, chronic myeloid leukemia cells K562 and Human normal hepatocyte are selected in this experiment
3 kinds of human cell's strains such as L-O2.
All cell strains cultivate containing the small ox blood of 10wt%, 100U/mL penicillin, 100U/mL streptomysin RPMI-
In 1640 culture medium, 37 DEG C of 5%CO containing volumetric concentration are set2It is cultivated in incubator.
2, the preparation of untested compound
Purity >=95% of test medicine used is configured to 200 μ after diluting its DMSO liquid storage with physiological buffer
The whole solution of mol/L, wherein final concentration≤1% of cosolvent DMSO, it is raw to various tumour cells to test compound under the concentration
Long inhibition level.
3, cell growth inhibition test (mtt assay)
(1) tumour cell of logarithmic growth phase is matched after trypsin digestion with the culture solution containing 10% calf serum
The cell suspension that concentration is 5000/mL is made, is inoculated in 96 well culture plates with every 190 μ L of hole, makes cell density to be measured extremely
1000~10000 holes (edge hole is filled with sterile PBS)；
(2) 5%CO2, 37 DEG C are incubated for for 24 hours, until cell monolayer is paved with bottom hole, the drug 10 of a certain concentration gradient is added in every hole
μ L, each concentration gradient set 4 multiple holes；
(3) 5%CO2, 37 DEG C are incubated for 48 hours, observe under inverted microscope；
(4) the MTT solution (5mg/mL PBS, i.e. 0.5%MTT) of 10 μ L is added in every hole, continues to cultivate 4h；
(5) culture is terminated, culture solution in hole is carefully sucked, the DMSO that 150 μ L are added in every hole sufficiently dissolves first a ceremonial jade-ladle, used in libation precipitating, vibration
It swings after device mixes, with wavelength is 570nm in microplate reader, reference wavelength is the OD value that 450nm measures each hole；
(6) it is arranged zeroing hole (culture medium, MTT, DMSO) simultaneously, (the drug dissolution of cell, same concentrations is situated between control wells
Matter, culture solution, MTT, DMSO).
(7) according to the OD value (OD value) measured, to judge living cells quantity, OD value is bigger, and cell activity is stronger.Benefit
Use formula：
Compound is calculated to the inhibiting rate of each cell strain growth, result is as shown in the following Table 1.
Table 1：IC of each compound to different cell strains50It is worth (μM)
Note：Experimental data is the average value of 3 experiments, and Nd indicates that test-compound lives to this cell detection less than inhibition
Property.
Claims (10)
1. enoxolone-hydrogen sulfide donor reagent derivatives with structure shown in the following general formula (I) or its is pharmaceutically acceptable
Salt：
Wherein,
When R isWhen, n 8；
When R isWhen, n 2,4,6 or 8；
When R isWhen, n is 6 or 8.
2. enoxolone described in claim 1-hydrogen sulfide donor reagent derivatives synthetic method, it is characterised in that：Extracting liquorice
Hypo acid, α, ω-two bromoalkane and alkali react in aprotic polar solvent, obtain compound 1；Take compound 1, hydrogen sulfide donor
Reagent and alkali react in aprotic polar solvent, obtain object crude product；Wherein, reaction is under conditions of being heated or not heated
It carries out, the structural formula of compound 1 is as follows：
Wherein, 2,4,6 or 8 n.
3. synthetic method according to claim 2, it is characterised in that：Include the following steps：
1) extracting liquorice hypo acid, α, ω-two bromoalkane and alkali react in aprotic polar solvent, and gained reactant removes solvent,
Residue is scattered in ethyl acetate, methylene chloride or ether, washed, after anhydrous sodium sulfate dries, filters, collects filtrate,
Compound 1 is obtained after filtrate concentration；
2) compound 1, hydrogen sulfide donor reagent and alkali is taken to react in aprotic polar solvent, gained reactant removes solvent,
Residue is scattered in ethyl acetate, methylene chloride or ether, washed, after anhydrous sodium sulfate dries, filters, collects filtrate,
Object crude product is obtained after filtrate concentration.
4. synthetic method according to claim 2 or 3, it is characterised in that：Gained compound 1 is through silica gel thin-layer chromatography or silicon
Subsequent operation is used further to after gel column chromatography eluting.
5. synthetic method according to claim 2 or 3, it is characterised in that：It further include purifying object crude product
Step：Object crude product obtained is specifically subjected to silica gel thin-layer chromatography or silica gel column chromatography, obtains object after purification.
6. synthetic method according to claim 2 or 3, it is characterised in that：The α, ω-two bromoalkane are 1,2- dibromo
Ethane, 1,3- dibromopropane, 1,4- dibromobutane, pentamethylene bromide, 1,6- dibromo-hexane, 1,7- dibromo-heptane or 1,8- bis-
Bromooctane.
7. synthetic method according to claim 2 or 3, it is characterised in that：The alkali is potassium carbonate, triethylamine, carbonic acid
Sodium, sodium bicarbonate, saleratus or cesium carbonate；The aprotic polar solvent is N,N-dimethylformamide, toluene and pyrrole
The combination of one or more of pyridine.
8. synthetic method according to claim 2 or 3, it is characterised in that：The hydrogen sulfide donor reagent is 5- to hydroxyl
Base phenyl -1,2- dithiole -3- thioketones, (R)-lipoic acid or 4- hydroxythiobenzamide.
9. synthetic method according to claim 2 or 3, it is characterised in that：In compound 1, hydrogen sulfide donor reagent and alkali
Reaction in, be added catalyst potassium iodide.
10. enoxolone described in claim 1-hydrogen sulfide donor reagent derivatives are in the drug for the treatment of chronic myelogenous leukemia
In application.
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