Summary of the invention
The object of the invention is to overcome deficiency of the prior art, find the active high class anti-androgen receptor compound of a class, a kind of tanshinone compound is provided, can be used as anti-androgen receptor medicine and be used for the treatment of the disease relevant to male sex hormone, can be used for treatment prostate cancer, as hyperplasia of prostate, acne, alopecia, hirsutism, muscle depletion, gonad function is weak, cholesterol is too high, male sterility, the diseases such as male's sexual is bad, anaemia, obesity.
Main purpose of the present invention is to provide chemical structure and the medical usage of this compounds.
Tanshinone compound, has the Compound I or II of following general structure:
Wherein R is CHO, Br, NO
2, OCH
3, CH
2oOCH
3, COCH
3, C
1-C
3carboxylic acid and sodium salt, carboxylicesters, vinyl cyanide, vinylformic acid and sodium salt or acrylate etc.
The purposes of tanshinone compound, for the preparation of anti-androgen receptor medicine.Be used for the treatment of the disease relevant to male sex hormone, can be used for treatment prostate cancer, as hyperplasia of prostate, acne, alopecia, hirsutism, muscle depletion, gonad function is weak, cholesterol is too high, male sterility, the diseases such as male's sexual is bad, anaemia, obesity.
Compound involved in the present invention is by following route synthetic method one:
The preparation method of tanshinone compound, step specific as follows: tanshinone IIA and Vilsmeier reagent (dimethyl formamide and phosphorus oxychloride) react prepares aldehyde compound, potassium permanganate can be adopted under phase-transfer catalyst effect to generate carboxylic acid, obtain carboxylicesters through esterification, react to sodium ethylate and generate corresponding sodium salt.
The preparation method of tanshinone compound, specifically carries out in accordance with the following steps:
(1) in reaction flask, add tanshinone IIA (1), dimethyl formamide (DMF), phosphorus oxychloride, 1-5 hour is reacted at 30-100 DEG C, reaction solution is poured in frozen water, has yellow mercury oxide, filters, washing, dry crude product, crude product obtains tanshinone IIA formaldehyde (2) through silica gel column chromatography, and wherein the mol ratio of reactant tanshinone IIA, dimethyl formamide (DMF), phosphorus oxychloride is 1:2-15:2-25, temperature of reaction 30-100 DEG C, reaction 1-5 hour.
(2) in reaction flask, 2 are added, acetone, Tetrabutyl amonium bromide and mass concentration are the potassium permanganate solution of 5%, at 20-80 DEG C, reaction 10-25 hour, solids removed by filtration, filtrate regulates PH=2 with hydrochloric acid, precipitation is had to generate, filter, dry, 1 is obtained with silica gel column chromatography, 6, 6-trimethylammonium-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furyl-2-formic acid (3), wherein reactant tanshinone IIA (1), Tetrabutyl amonium bromide, the potassium permanganate of 5%, acetone mol ratio is 1:0.05-0.2:2-5:5-25, temperature of reaction 20-80 DEG C, reaction 10-25 hour.
(3) in reaction flask, 1 is added, 6, 6-trimethylammonium-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furyl-2-formic acid (3), dehydrated alcohol, 10% alcohol sodium solution, namely solid is had to separate out, filter to obtain sodium salt 1, 6, 6-trimethylammonium-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furyl-2-sodium formiate (4), wherein 1, 6, 6-trimethylammonium-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furyl-2-formic acid (3), dehydrated alcohol, 10% alcohol sodium solution mol ratio is 1:2-5:2-15:0.95-1.25, temperature of reaction 20-80 DEG C, reaction 1-3 hour.
(4) 1,6,6-trimethylammonium-6 is added in reaction flask, 7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-formic acid (3), dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride, at room temperature react 2-12 hour, solids removed by filtration, obtains 1 with silica gel column chromatography, and 6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-formic acid ester compound (5).Wherein reactant 1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-formic acid (3), dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride equimolar ratio be 1:1-3:0.1-0.25:1-10:5-50, temperature of reaction 0-80 DEG C, reaction 1-12 hour.
Compound involved in the present invention is by following route synthetic method two
The preparation method of tanshinone compound, specifically carries out in accordance with the following steps:
Tanshinone IIA and nitration mixture (acetic acid and nitric acid) carry out nitration reaction and obtain 2-nitro-1, 6, 6-trimethylammonium-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans (6), tanshinone IIA (1) is added in reaction flask, diacetyl oxide, glacial acetic acid and nitrosonitric acid, react after 1-8 hour and stop pouring in frozen water, separate out a large amount of yellow solid, filter, washing, dry, obtain nitro-compound 2-nitro-1, 6, 6-trimethylammonium-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans (6), wherein reactant tanshinone IIA (1), diacetyl oxide, glacial acetic acid and nitrosonitric acid mol ratio are 1:1-2:2-5:2-5, temperature of reaction-10 is to 0 DEG C, reaction 1-12 hour.
Tanshinone IIA and N-chlorosuccinimide (NCS), N-bromosuccinimide (NBS), N-iodosuccinimide (NIS) react and generate corresponding halogenated product 2-halogen-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (6).
In reaction flask, add tanshinone IIA (1), acetic acid, N-halosuccinimides, acetic acid, at temperature of reaction-10 is to 20 DEG C, reacts 2-10 hour, adds water, 2-halogen-1,6,6-trimethylammonium-6 is obtained with dichloromethane extraction, 7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (7), wherein reactant tanshinone IIA (1), N-halosuccinimides, acetic acid mol ratio are 1:1-3:2-15, and temperature of reaction-10, to 20 DEG C, reacts 2-10 hour.
Tanshinone IIA generates acetyl compound 8 with excess acetyl chloride under catalyst action.
Tanshinone IIA (1), pyridine, Acetyl Chloride 98Min. is added in reaction flask; react 1-5 hour at 20-100 DEG C after, reaction solution is poured in frozen water; filter; washing, dry crude product, crude product obtains acetyl compound 2-ethanoyl-1 through silica gel column chromatography; 6; 6-trimethylammonium-6,7,8; 9; 10,11-six hydrogen-10,11-dioxo phenanthro-[1; 2-b] furans (8); wherein reactant tanshinone IIA (1), pyridine, Acetyl Chloride 98Min. mol ratio are 1:2-25:1-5, temperature of reaction 20 to 100 DEG C, reaction 1-5 hour.
Compound involved in the present invention is by following route synthetic method three
Method three of the present invention step specific as follows:
Tanshinone IIA is oxidized and generates aldehyde compound under tin anhydride catalysis, carries out reduction oxy-compound respectively, generates acrylic compounds, carry out esterification obtain corresponding acrylic ester compound to alcohol with propanedioic acid under pyridine and piperidines catalysis.
(1) in reaction flask, add tanshinone IIA (1), tin anhydride, dioxane, heating reflux reaction 5-24 hour, filter, concentrated, obtain 6,6-dimethyl-6 with silica gel column chromatography, 7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans-1-formaldehyde (9), wherein reactant tanshinone IIA (1), tin anhydride, dioxane mol ratio are 1:0.5-2.5:3-25, temperature of reaction 40 to 110 DEG C, reaction 5-24 hour.
(2) in reaction flask, 6 are added, 6-dimethyl-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans-1-formaldehyde (9), methyl alcohol, methylene dichloride, sodium borohydride, 10-120 minute is reacted at-10 to 5 DEG C, be extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration, 1-(methylol is obtained with silica gel column chromatography)-6, 6-dimethyl-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans (10), wherein 6, 6-dimethyl-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans-1-formaldehyde (9), methyl alcohol, methylene dichloride, sodium borohydride mol ratio is 1:2-10:2-5:1-2.5, temperature of reaction-10 is to 5 DEG C, reaction 10-120 minute.
(3) in reaction flask, 1-(methylol is added)-6, 6-dimethyl-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans (10), methylene dichloride, triethylamine, diacetyl oxide, reaction response 1-3 hour at-10 to 5 DEG C, be extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration, 6 are obtained with silica gel column chromatography, 6-dimethyl-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans-2-methyl acetic acid ester (11), wherein 1-(methylol)-6, 6-dimethyl-6, 7, 8, 9, 10, 11-six hydrogen-10, 11-dioxo phenanthro-[1, 2-b] furans (10), methylene dichloride, triethylamine, diacetyl oxide mol ratio is 1:2-10:1-2.5:1-2.5, temperature of reaction-10 is to 5 DEG C, reaction 1-3 hour.
Compound involved in the present invention is by following route synthetic method four
Method four of the present invention step specific as follows:
TANSHINONES aldehyde compound carries out selective reduction reduction reaction and generates oxy-compound or ether compound respectively, and oxy-compound and acetic anhydride generate corresponding ester compound.
(1) in reaction flask, add tanshinone IIA formaldehyde (2), methyl alcohol, methylene dichloride, sodium borohydride, at-10 to 5 DEG C, reaction times 10-60 minute, be extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration obtains compound 12, and wherein the mol ratio of tanshinone IIA formaldehyde (2), methyl alcohol, methylene dichloride, sodium borohydride is 1:10-100:10-100:1-2.5, temperature of reaction-10 to 5 DEG C, reaction times 10-60 minute.
(2) in reaction flask, compound 12, methylene dichloride, diacetyl oxide is added, at-10 to 5 DEG C, reaction 1-3 hour, be extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration, compound 13 is obtained with silica gel column chromatography, wherein the mol ratio of compound 12, methylene dichloride, diacetyl oxide is 1:10-100:1-2.5, temperature of reaction-10 to 5 DEG C, reaction times 1-3 hour.
(3) in reaction flask, add tanshinone IIA formaldehyde (2), 10%(mass concentration) palladium carbon, methyl alcohol, at room temperature hydrogenation reaction 20-24 hour, filter palladium carbon, vacuum concentration, target compound 14 is obtained with silica gel column chromatography, wherein the mol ratio of compound tanshinone IIA formaldehyde (2), 10% palladium carbon, methyl alcohol, hydrogen is 1:0.01-0.2:10-100:1:5, temperature of reaction 10 to 40 DEG C, reaction times 10-36 hour.
Compound involved in the present invention is by following route synthetic method five
Method five of the present invention step specific as follows:
TANSHINONES carbonyl compound, under pyridine and piperidines catalysis, carry out decarboxylation fertilizer with propanedioic acid should generation acrylic compounds, then carries out esterification to alcohol and obtain corresponding acrylic ester compound.
(1) in reaction flask, compound 15, benzene, propanedioic acid, pyridine is added, reflux divides water, and reaction 8-20 hour, removes solvent under reduced pressure, the aqueous sodium carbonate of 5% is added in residue, filter, filtrate transfers in about PH=2 with concentrated hydrochloric acid, filters, carboxylic acid cpd 16 is obtained with silica gel column chromatography, wherein compound 15, benzene, propanedioic acid, pyridine equimolar ratio are 1:10-30:1-5:0.2-2.5, temperature of reaction 40-100 DEG C, reaction 8-20 hour.
(2) in reaction flask, add carboxylic acid cpd 16, dehydrated alcohol, 10%(mass concentration) alcohol sodium solution, namely solid is had to separate out, filter to obtain sodium salt 18, wherein reactant 16, dehydrated alcohol, 10% alcohol sodium solution mol ratio are 1:2-5:2-5:0.95-1.25, temperature of reaction 20-80 DEG C, reaction 1-3 hour.
(3) carboxylic acid cpd 16, dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride is added in reaction flask, at room temperature react 2-12 hour, solids removed by filtration, obtains ester cpds 17 with silica gel column chromatography.Wherein reactant 16, dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride equimolar ratio are 1:1-3:0.1-0.25:1-3:5-50, temperature of reaction 0-80 DEG C, reaction 1-12 hour.
Compound involved in the present invention is by following route synthetic method six
Method six of the present invention step specific as follows:
TANSHINONES is reacted to ethyl chloroacetate or chloropropionic acid ethyl ester and is generated corresponding carboxylic acid ester compound under zirconium chloride catalysis, then hydrolysis generates corresponding carboxylic acid cpd in the basic conditions, reacts generate corresponding sodium salt to sodium ethylate.
(1) in reaction flask, add tanshinone IIA (1), methylene dichloride, zirconium tetrachloride, ethyl chloroacetate (or chloropropionate), return stirring reaction 2-10 hour, solids removed by filtration, filter vacuum concentrates, residual silica gel column chromatography obtains compound 19, wherein tanshinone IIA (1), methylene dichloride, zirconium tetrachloride, ethyl chloroacetate (or chloropropionate) mol ratio are 1:15-50:2-5:1-3, temperature of reaction 20-80 DEG C, reaction 2-10 hour.
(2) in reaction flask, add to obtain compound 19,5%(mass concentration) aqueous sodium hydroxide solution, reflux 1-4 hour, filters, and filtrate regulates about PH2 with concentrated hydrochloric acid, separates out solid, filters, dry, obtains carboxylic acid cpd 20 with silica gel column chromatography,
(3) wherein reactant compound 19 and 5%(mass concentration) aqueous sodium hydroxide solution mol ratio be 1:1-2.5, temperature of reaction 30-110 DEG C, reaction 1-4 hour.
(4) in reaction flask, add carboxylic acid cpd 20, dehydrated alcohol, 10% alcohol sodium solution, namely solid is had to separate out, filter to obtain sodium salt 21, wherein reactant 20, dehydrated alcohol, 10%(mass concentration) alcohol sodium solution mol ratio is 1:2-5:2-5:0.95-1.25, temperature of reaction 20-80 DEG C, reaction 1-3 hour.
(5) carboxylic acid cpd 20, dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride is added in reaction flask, at room temperature react 2-12 hour, solids removed by filtration, obtains ester cpds 22 with silica gel column chromatography.Wherein reactant 20, dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride equimolar ratio are 1:1-3:0.1-0.25:1-3:5-50, temperature of reaction 0-80 DEG C, reaction 1-12 hour.
Compound involved in the present invention is by following route synthetic method seven
Method seven of the present invention step specific as follows:
TANSHINONES is reacted with butyryl oxide and is generated butyric acid compound under zirconium chloride catalysis, then reacts with alcohol and generate carboxylic acid ester compound, reacts generate corresponding sodium salt to sodium ethylate.
(1) in reaction flask, add tanshinone IIA (1), methylene dichloride, zirconium tetrachloride, Succinic anhydried, return stirring reaction 2-12 hour, solids removed by filtration, filter vacuum concentrates, residual silica gel column chromatography obtains carboxylic acid cpd 23, wherein reactant tanshinone IIA (1), methylene dichloride, zirconium tetrachloride, Succinic anhydried mol ratio are 1:15-100:2-5:1-1.5, temperature of reaction 20-80 DEG C, reaction 2-12 hour.
(2) in reaction flask, add carboxylic acid cpd 23, dehydrated alcohol, 10%(mass concentration) alcohol sodium solution, namely solid is had to separate out, filter to obtain sodium salt 24, wherein reactant 23, dehydrated alcohol, 10% alcohol sodium solution mol ratio are 1:2-5:2-5:0.95-1.25, temperature of reaction 20-80 DEG C, reaction 1-3 hour.
(3) carboxylic acid cpd 23, dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride is added in reaction flask, at room temperature react 2-12 hour, solids removed by filtration, obtains ester cpds 25 with silica gel column chromatography.Wherein reactant 23, dicyclohexylcarbodiimide (DCC), DMAP (DMAP), absolute alcohol, methylene dichloride equimolar ratio are 1:1-3:0.1-0.25:1-3:5-50, temperature of reaction 0-80 DEG C, reaction 1-12 hour.
Advantage of the present invention:
AR has multiple hypotype, perfect form (fAR) and spliced variants type androgen receptor (ARVs), wherein spliced variants type androgen receptor (ARVs) is that advanced prostate cancer is distinctive, we all effectively suppress perfect form (fAR) and spliced variants type androgen receptor (ARVs) by the tanshinone compound of invention, can be used for the treatment of advanced prostate cancer.Compared with existing clinical medicine bicalutamide, perfect form (fAR) can only be suppressed, spliced variants type androgen receptor (ARVs) can not be suppressed, cause bicalutamide invalid to advanced prostate cancer, the present invention overcomes the deficiency of upper existing androgen antagonist medicine, can be used for treating advanced prostate cancer.
Embodiment
The present invention relates to the medicine of tanshinone compound, its structural formula of the compound comprised is as follows:
Example one, 1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans-2-formaldehyde (Tan-001)
Tanshinone IIA (2.94g is added in reaction flask, 19mmol), 50ml dimethyl formamide (DMF), stirred at ambient temperature drips 7ml phosphorus oxychloride, at 70-80 DEG C, react reaction solution after 2 hours pours in the frozen water of 750ml, there is yellow mercury oxide, filter, washing, dry crude product, crude product obtains through silica gel column chromatography, productive rate 67%
1h-NMR (300MHz, (CDCl
3)), δ: 9.86 (s, 1H), 8.80 (d, J=6.3Hz, 1H), 7.71 (d, J=6.3Hz, 1H), 3.21 (m, 2H), 2.65 (s, 3H), 1.78-1.82 (m, 2H), 1.65-1.69 (m, 2H), 1.42 (s, 6H).
Bromo-10,11-dioxo-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen phenanthro-[1, the 2-b] furans (Tan-002) of example two, 2-
In reaction flask, add tanshinone IIA (1.47g, 5mmol), acetic acid 80ml, slowly drip N-bromosuccinimide NBS(1.07g, solution 6mmol) in 25ml acetic acid, room temperature reaction 6 hours, adds water, with dichloromethane extraction, combining extraction liquid, washes with water successively, and sodium sulfite aqueous solution washs, saturated sodium-chloride water solution washs, anhydrous sodium sulfate drying, obtains red solid with silica gel column chromatography
1h-NMR (300MHz, (CDCl
3)), δ: 7.68 (d, J=7.9Hz, 1H), 7.53(d, J=7.9Hz, 1H), 3.19 (m, 2H), 2.21 (s, 3H), 1.77-1.83 (m, 2H), 1.64-1.71 (m, 2H), 1.31 (s, 6H).
Chloro-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (Tan-003) of example three, 2-
Reaction, with example two, replaces NBS with N-chlorosuccinimide (NCS), obtains target product,
1h-NMR (300MHz, (CDCl
3), δ: 7.69 (d, J=7.2Hz, 1H), 7.56 (d, J=7.2Hz, 1H), 3.19 (m, 2H), 2.23 (s, 3H), 1.77-1.85 (m, 2H), 1.64-1.75 (m, 2H), 1.31 (s, 6H).
Iodo-10,11-dioxo-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen phenanthro-[1, the 2-b] furans (Tan-004) of example four, 2-
In reaction flask, add tanshinone IIA (59mg, 0.2mmol), methylene dichloride 20ml, be cooled to-40 DEG C, slowly drip N-iodosuccinimide NIS(67.2g, 0.6mmol) 5ml dichloromethane solution ,-40 DEG C of reactions 6 hours, add water, with dichloromethane extraction, combining extraction liquid, wash with water successively, sodium sulfite aqueous solution washs, saturated sodium-chloride water solution washs, and anhydrous sodium sulfate drying, obtains red solid with silica gel column chromatography.
1H-NMR(300MHz,(CDCl
3),δ:7.63(d,J=7.5Hz,1H),7.55(d,J=7.5Hz,1H),3.18(m,2H),2.23(s,3H),1.78-1.86(m,2H),1.65-1.72(m,2H),1.32(s,6H)。
Example five, 2-nitro-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (Tan-005)
Tanshinone IIA (59mg is added in reaction flask, 0.2mmol), 30ml diacetyl oxide, be cooled to less than 0 DEG C with ice-water bath, add in the nitration mixture that glacial acetic acid (1.4g) and nitrosonitric acid (1.5g) make, react stopped reaction after 3-4 hour, reaction mixture is poured in frozen water, separate out a large amount of yellow solid, filtration, washing, drying, obtain target compound with silica gel column chromatography, yield 68%
1h-NMR (300MHz, (CDCl
3)), δ: 7.79 (d, J=7.2Hz, 1H), 7.74 (d, J=7.2Hz, 1H), 3.21 (m, 2H), 2.76 (s, 3H), 1.78-1.86 (m, 2H), 1.66-1.73 (m, 2H), 1.32 (s, 6H).
Example six, 2-(methylol)-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (Tan-006)
Tan-001(129mg is added in reaction flask, 0.4mmol), 15ml methyl alcohol and 10ml methylene dichloride, be cooled to less than 0 DEG C with ice-water bath, add sodium borohydride (30mg, 0.81mmol), under ice bath, stirring reaction 15-20 minute, is extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration, target compound is obtained, yield 75% with silica gel column chromatography
1h-NMR (300MHz, (CDCl
3)), δ: 7.60 (d, J=6.8Hz, 1H), 7.51 (d, J=6.8Hz, 1H), 4.68 (s, 2H), 3.16 (m, 2H), 2.26 (s, 3H), 1.76-1.85 (m, 2H), 1.63-1.76 (m, 2H), 1.31 (s, 6H).
Example seven, 2-ethanoyl-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (Tan-007)
In reaction flask, add tanshinone IIA (2.94g, 19mmol), 120ml pyridine, stirred at ambient temperature drips 8ml Acetyl Chloride 98Min., at 70-80 DEG C, react reaction solution after 2 hours pours in the frozen water of 750ml, filters, washing, dry crude product, crude product obtains target compound through silica gel column chromatography, productive rate 73%
1h-NMR (300MHz, (CDCl
3)), δ:, 8.09 (d, J=7.2Hz, 1H), 7.63 (d, J=7.2Hz, 1H), 3.19 (m, 2H), 2.89 (s, 3H), 2.35 (s, 3H), 1.76-1.85 (m, 2H), 1.66-1.78 (m, 2H), 1.38 (s, 6H)
Example eight, 1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans is muttered-2-methyl acetic acid ester (Tan-008)
Tan-006(130mg is added in reaction flask, 0.4mmol), 25ml methylene dichloride, be cooled to less than 0 DEG C with ice-water bath, drip diacetyl oxide (1ml), stirring reaction 1-1.5 hour under ice bath, be extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration, obtains target compound with silica gel column chromatography, yield 86%
1h-NMR (300MHz, (CDCl
3), δ: 7.61 (d, J=7.2Hz, 1H), 7.54 (d, J=7.2Hz, 1H), 5.07 (s, 2H), 3.14 (m, 2H), 2.28 (s, 3H), 2.02 (s, 3H), 1.78 (m, 2H), 1.63 (m, 2H), 1.29 (s, 6H).
Example nine, 2-(methoxyl group)-1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (Tan-009)
In reaction flask, add Tan-001(260mg, 0.8mmol), 10% palladium carbon (30mg), 50ml methyl alcohol, at room temperature hydrogenation reaction 20-24 hour, filter palladium carbon, vacuum concentration, obtains target compound with silica gel column chromatography, yield 29%,
1h-NMR (300MHz, (CDCl
3), δ: 7.67 (m, 2H), 4.08 (s, 3H), 3.18 (m, 2H), 2.39 (s, 3H), 1.76-1.79 (m, 2H), 1.61-1.65 (m, 2H), 1.29 (s, 6H).
Example ten, 6,6-dimethyl-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans is muttered-1-formaldehyde (Tan-010)
In reaction flask, add tanshinone IIA (200mg, 0.68mmol), tin anhydride (225mg, 2.03mmol), 25ml dioxane, heating reflux reaction 10-12 hour, filter, concentrated, obtain target compound Tan-010 with silica gel column chromatography, yield 33%,
1h-NMR (300MHz, (CDCl
3), δ: 10.33 (s, 1H), 8.01 (d, J=7.5Hz, 1H), 7.69 (d, J=7.5Hz, 1H), 3.23 (m, 2H), 1.76-1.88 (m, 2H), 1.63-1.73 (m, 2H), 1.32 (s, 6H).
Example 11,1-(methylol)-6,6-dimethyl-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans (Tan-011)
Tan-010(125mg is added in reaction flask, 0.4mmol), 10ml methyl alcohol and 20ml methylene dichloride, be cooled to less than 0 DEG C with ice-water bath, add sodium borohydride (29mg, 0.80mmol), under ice bath, stirring reaction 30-40 minute, is extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration, target compound is obtained, yield 67% with silica gel column chromatography
1h-NMR (300MHz, (CDCl
3)) δ: 7.68 (d, J=7.8Hz, 1H), 7.57 (d, J=7.2Hz, 1H), 7.37 (s, 1H), 4.25 (s, 2H), 3.19 (m, 2H), 1.78-1.83 (m, 2H), 1.64-1.72 (m, 2H), 1.31 (s, 6H).
Example 12,6,6-dimethyl-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furans-2-methyl acetic acid ester (Tan-012)
Tan-011(125mg is added in reaction flask, 0.4mmol), 20ml methylene dichloride, be cooled to less than 0 DEG C with ice-water bath, drip 2ml triethylamine, then 1ml diacetyl oxide is dripped, in ice bath, stirring reaction 1 hour, is extracted with ethyl acetate, washing, anhydrous sodium sulfate drying, vacuum concentration, target compound is obtained, yield 83% with silica gel column chromatography
1h-NMR (300MHz, (CDCl
3)) δ: 7.59 (d, J=6.0Hz, 1H), 7.55 (d, J=6.0Hz, 1H), 7.473 (s, 2H), 5.24 (s, 2H), 3.19 (t, 2H), 2.11 (s, 3H), 1.76-1.81 (m, 2H), 1.64-1.67 (m, 2H), 1.31 (s, 6H).
Example 13,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-formic acid (Tan-013)
Tan-001(258mg is added in reaction flask, 0.8mmol), 40ml acetone, Tetrabutyl amonium bromide (15mg), be cooled to less than 0 DEG C with ice-water bath, drip concentration be 5% potassium permanganate solution 15ml, at room temperature react 15-20 hour, solids removed by filtration, filtrate regulates PH=2 with hydrochloric acid, has precipitation to generate, filtration, drying, target compound is obtained, yield 46% with silica gel column chromatography
1h-NMR (400MHz, (CDCl
3), δ: 9.89 (s, 1H), 7.99 (d, J=7.8Hz, 1H), 7.67 (d, J=7.8HZ, 1H), 2.97 (t, 7.8HZ, 7.8Hz, 2H), 2.68 (s, 3H), 2.11 (t, J=7.8HZ, 7.8HZ, 2H), 1.81 (m, 2H), 1.37 (s, 6H).
Example 14,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-methyl acetate (Tan-014)
Tan-013(100mg is added in reaction flask, 0.3mmol), dicyclohexylcarbodiimide (DCC) ((80mg, 0.38mmol), (DMAP (DMAP) (10mg), 1ml anhydrous methanol, 25ml methylene dichloride, at room temperature react 4-6 hour, solids removed by filtration, target compound is obtained, yield 82%, ESI-MS [M with silica gel column chromatography
+] (m/z): 352.13.
Example 15,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-ethyl formate (Tan-015)
Reaction, with example 15, replaces anhydrous methanol with dehydrated alcohol, obtains target product, yield 74%, ESI-MS [M
+] (m/z): 366.15.
Example 16,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-sodium formiate (Tan-016)
Tan-013(300mg, 0.89mmol is added in reaction flask) be dissolved in dehydrated alcohol, add 10% alcohol sodium solution of equivalent, namely have solid to separate out, filter to obtain target product, yield 91%, ESI-MS [M
+] (m/z): 360.14.
Example 17,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-ethyl acetate (Tan-017)
Tanshinone IIA (1.47g is added in reaction flask, 5mmol), 75ml methylene dichloride, zirconium tetrachloride (1.2g, 5.2mmol), 3ml ethyl chloroacetate, return stirring reaction 4-6 hour, solids removed by filtration, filter vacuum concentrates, and residual silica gel column chromatography obtains target compound, yield 79%, ESI-MS [M
+] (m/z): 380.16.
Example 18,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-acetic acid (Tan-018)
Tan-017(760mg is added in reaction flask, 2mmol), the aqueous sodium hydroxide solution (150ml) of 5%, reflux 1-2 hour, stopped reaction, filter, filtrate regulates about PH2 with concentrated hydrochloric acid, separates out solid, filters, dry, target compound is obtained, yield 81%, ESI-MS [M with silica gel column chromatography
+] (m/z): 352.13.
Example 19,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-methyl acetate (Tan-019)
Reaction, with example 15, replaces Tan-013 with Tan-019, obtains target product, yield 84%, ESI-MS [M
+] (m/z): 366.15.
Example 20,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-sodium acetate (Tan-020)
Tan-018(300mg, 0.85mmol is added in reaction flask) be dissolved in dehydrated alcohol, add 10% alcohol sodium solution of equivalent, namely have solid to separate out, filter to obtain target product, yield 89%, ESI-MS [M
+] (m/z): 374.11.
Example 21,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-ethyl propionate (Tan-021)
Tanshinone IIA (1.47g is added in reaction flask, 5mmol), 75ml methylene dichloride, zirconium tetrachloride (1.2g, 5.2mmol), 3ml chloropropionate, return stirring reaction 4-6 hour, solids removed by filtration, filter vacuum concentrates, and residual silica gel column chromatography obtains target compound, yield 63%, ESI-MS [M
+] (m/z): 394.18.
Example 22,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-propionic acid (Tan-022)
Tan-021(800mg is added in reaction flask, 2mmol), the aqueous sodium hydroxide solution (150ml) of 5%, reflux 1-2 hour, stopped reaction, filter, filtrate regulates about PH2 with concentrated hydrochloric acid, separates out solid, filters, dry, target compound is obtained, yield 72%, ESI-MS [M with silica gel column chromatography
+] (m/z): 366.15.
Example 23,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-methyl propionate (Tan-023)
Reaction, with example 15, replaces Tan-013 with Tan-021, obtains target product, yield 86%, ESI-MS [M
+] (m/z): 380.16.
Example 24,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-Sodium Propionate (Tan-024)
Tan-021(300mg, 0.82mmol is added in reaction flask) be dissolved in dehydrated alcohol, add 10% alcohol sodium solution of equivalent, namely have solid to separate out, filter to obtain target product, yield 88%, ESI-MS [M
+] (m/z): 388.13.
Example 25,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-butyric acid (Tan-025)
Tanshinone IIA (1.47g is added in reaction flask, 5mmol), 100ml methylene dichloride, zirconium tetrachloride (1.2g, 5.2mmol), Succinic anhydried (0.69g, 7.5mmol), return stirring reaction 4-6 hour, solids removed by filtration, filter vacuum concentrates, residual silica gel column chromatography obtains target compound, yield 75%, ESI-MS [M
+] (m/z): 380.16.
Example 26,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-methyl-butyrate (Tan-026)
Reaction, with example 15, replaces Tan-013 with Tan-025, obtains target product, yield 86%, ESI-MS [M
+] (m/z): 394.18.
Example 27,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-ethyl butyrate (Tan-027)
Reaction, with example 15, replaces Tan-013 with Tan-025, and dehydrated alcohol replaces anhydrous methanol, obtains target product, yield 83%, ESI-MS [M
+] (m/z): 408.19.
Example 28,1,6,6-trimethylammonium-6,7,8,9,10,11-six hydrogen-10,11-dioxo phenanthro-[1,2-b] furyl-2-Sodium propanecarboxylate (Tan-028)
Tan-025(300mg, 0.79mmol is added in reaction flask) be dissolved in dehydrated alcohol, add 10% alcohol sodium solution of equivalent, namely have solid to separate out, filter to obtain target product, yield 89%, ESI-MS [M
+] (m/z): 402.14.
Example 29 3-(6,7,8,9,10,11-six hydrogen)-1,6,6-trimethylammonium-10,11-dioxo phenanthro-[1,2-b] furans-2-base-2-vinylformic acid (Tan-029)
(Tan-001) (1.47g is added in reaction flask, 5mmol), 100ml benzene, propanedioic acid (1.2g, 5.2mmol), 10ml pyridine, reflux divides water, reaction 10-12 hour, remove solvent under reduced pressure, in residue, add the aqueous sodium carbonate of 5%, filter, filtrate transfers in about PH=2 with concentrated hydrochloric acid, filter, obtain target compound with silica gel column chromatography, yield 76%.
1H-NMR(300MHz,(DMSO-d
6),δ:12.56(s,1H),7.78-7.85(m,2H),7.51(d,1H),6.46(d,1H),3.09(m,2H),1.73(m,2H),1.73(m,2H),1.64(m,2H),2.37(s,3H),1.28(s,6H)。
Example 30 3-(6,7,8,9,10,11-six hydrogen)-1,6,6-trimethylammonium-10,11-dioxo phenanthro-[1,2-b] furans-2-base-2-vinylformic acid formic acid (Tan-030)
Reaction, with example 15, replaces Tan-013 with Tan-029, obtains target product, yield 81%.
1H-NMR(300MHz,(CDCl
3),δ:7.78(d,1H),7.65(d,1H),7.49(d,H),6.42(d,1H),3.82(s,3H),2.92(m,2H),2.40(s,3H),2.20(m,2H),1.34(s,6H)。
Example 31 3-(6,7,8,9,10,11-six hydrogen)-1,6,6-trimethylammonium-10,11-dioxo phenanthro-[1,2-b] furans-2-base-2-ethyl propenoate (Tan-031)
Reaction, with example 15, replaces Tan-013 with Tan-029, and dehydrated alcohol replaces anhydrous methanol, reacts to obtain target product, yield 85%.
1H-NMR(300MHz,(CDCl
3),δ:7.70(m,2H),7.46(d,1H),6.42(d,1H),5.39(s,1H),4.29(m,2H),3.29(m,2H),2.39(s,3H),1.79(m,2H),1.64(m,2H),1.27(m,3H),1.17(s,6H)。
Example 32 3-(6,7,8,9,10,11-six hydrogen)-1,6,6-trimethylammonium-10,11-dioxo phenanthro-[1,2-b] furans-2-base-2-sodium acrylate (Tan-032)
Tan-029(300mg, 0.83mmol is added in reaction flask) be dissolved in dehydrated alcohol, add 10% alcohol sodium solution of equivalent, namely have solid to separate out, filter to obtain target product, yield 81%, ESI-MS [M
+] (m/z): 386.11.
Example 33 3-[(6,7,8,9,10,11-six hydrogen)-1,6,6-tetramethyl--10,11-dioxo phenanthro-[1,2-b] furans-2-base-)]-2-butylene acid (Tan-033)
(Tan-007) (1.68g, 5mmol), 100ml benzene, propanedioic acid (1.2g, 5.2mmol), 10ml pyridine is added in reaction flask, reflux divides water, reaction 10-12 hour, removes solvent under reduced pressure, adds the aqueous sodium carbonate of 5% in residue, filter, filtrate transfers in about PH=2 with concentrated hydrochloric acid, filters, obtains target compound with silica gel column chromatography, yield 81%, ESI-MS [M
+] (m/z): 378.16.
Example 34 3-(6,7,8,9,10,11-six hydrogen)-1,6,6-trimethylammonium-10,11-dioxo phenanthro-[1,2-b] furans-2-base-) 2-methyl acrylate (Tan-034)
Reaction, with example 15, replaces Tan-013 with Tan-033, obtains target product, yield 79%, ESI-MS [M
+] (m/z): 392.17.
Example 35 3-(6,7,8,9,10,11-six hydrogen)-1,6,6-trimethylammonium-10,11-dioxo phenanthro-[1,2-b] furans-2-base-) 2-ethyl propenoate (Tan-035)
Reaction, with example 15, replaces Tan-013 with Tan-033, and dehydrated alcohol replaces anhydrous methanol, obtains target product, yield 75%, ESI-MS [M
+] (m/z): 406.19.
Example 36 3-[(6,7,8,9,10,11-six hydrogen)-1,6,6-tetramethyl--10,11-dioxo phenanthro-[1,2-b] furans-2-base-)]-2-butylene acid sodium (Tan-036)
Tan-033(300mg, 0.81mmol is added in reaction flask) be dissolved in dehydrated alcohol, add 10% alcohol sodium solution of equivalent, namely have solid to separate out, filter to obtain target product, yield 86%, ESI-MS [M
+] (m/z): 400.15.
The present invention utilizes the growth-inhibiting effect to the male hormone dependency Human Prostate Cancer Cells LNCaP of male sex hormone induction and the androgen independence Human Prostate Cancer Cells CWR22RV1 cell of Steroid-Resistance, measures the biologic activity of compound.
Compound biological activity determination
We check above-claimed cpd on the bioactive impact of androgen receptor with prostate cancer cell, and our cell has two kinds: LNCaP and CWR22RV1.LNCaP cell is male hormone dependency Human Prostate Cancer Cells, and CWR22RV1 is the androgen independence Human Prostate Cancer Cells of Steroid-Resistance.These two kinds of cells all express androgen receptor, and male sex hormone can induce the genetic expression of prostate specific antigen (PSA), and can induce the growth of this cell.The growth-inhibiting effect of cell, measures the biologic activity of compound.
Specific antigens is an important indicator of prostate cancer, and its expression is regulated and controled by androgen receptor, because of the physiologically active of the expression level reflection androgen receptor of a little prostate specific antigen.In order to check above-claimed cpd to the rejection ability of androgen receptor function, we effectively induce the expression of prostate specific antigen with male hormone DHT, then measure the rejection ability that compound is expressed the prostate specific antigen that DHT induces.We use the RPMI-1640 substratum (100 units/ml penicillin containing 10% foetal calf serum, 100mg/L Streptomycin sulphate and 10% foetal calf serum) cultivate prostate cancer cell LNCaP and CWR22RV1, before test compounds suppresses androgen receptor ability, I falls in the RPMI-1640 substratum of androgenic foetal calf serum (Charcoal-strippedFBS) to cultivate in containing 10% through wood charcoal adsorption at these two kinds of cell cultures of people, after two days, we add the DHT and 0.25 of androgenous 11 0nM in nutrient solution, 0.5, 1, 2, 5, 10 μMs, the test compounds of concentration tonsure, cultivate after four days, we are secreted into the concentration of the prostate specific antigen of nutrient solution with integrated enzyme reaction reagent measuring, then rejection ability (the IC that test compounds suppresses 50 percent of prostate specific antigen is calculated
50).Table 1 list tested compound to 50 percent of the prostate specific antigen of the androgen independence Human Prostate Cancer Cells CWR22RV1 of androgen-dependent Human Prostate Cancer Cells LNCaP and Steroid-Resistance rejection ability (IC
50).
Table 1, tested compound to 50 percent of the prostate specific antigen of the androgen independence Human Prostate Cancer Cells CWR22RV1 of androgen-dependent Human Prostate Cancer Cells LNCaP and Steroid-Resistance rejection ability (IC
50).
We also examine the growth-inhibiting effect of the androgen independence Human Prostate Cancer Cells CWR22RV1 cell of male hormone dependency Human Prostate Cancer Cells LNCaP that some compounds induce male sex hormone and Steroid-Resistance, similar with above method, we are placed on LNCaP cell and CWR22RV1 cell to fall in the RPMI-1640 substratum of androgenic foetal calf serum to cultivate after 2 days through wood charcoal adsorption containing 10%, add the test compounds of male hormone DHT and finite concentration gradient in the medium, its ultimate density is made to reach the DHT and 1 μM of 10nM, 2.5 μM, the test compounds of 5 μMs of concentration tonsures, cultivate after six days, we calculate the total cellular score of growth, the total cellular score that accompanying drawing display grows when being and deducting do not add DHT with the Growth of Cells sum of 10nMDHT induction is 100% growth, the compound of different concns is to the allometry rejection ability of the androgen independence human prostata cancer CWR22RV1 cell of male hormone dependency Human Prostate Cancer Cells LNCaP and Steroid-Resistance, three kinds of chemical structures are strong to the widely used bicalutamide of the growth-inhibiting energy force rate (Bicalutamide) of LNCaP and CWR22RV1.
Accompanying drawing, the allometry rejection ability of the androgen independence Human Prostate Cancer Cells CWR22RV1 (Fig. 2) of the male hormone dependency Human Prostate Cancer Cells LNCaP (Fig. 1) that three kinds of compounds are induced male sex hormone and Steroid-Resistance.
LNCaP and CWR22RV1 cell cultures is in falling in the RPMI-1640 substratum of androgenic foetal calf serum to cultivate after 2 days through wood charcoal adsorption containing 10%, add the test compounds of male hormone DHT and finite concentration gradient in the medium, its ultimate density is made to be the DHT and 1 μM of 10nM, 2.5 μM, the test compounds of 5 μMs of concentration tonsures, cultivate after six days, we calculate the total cellular score of growth, and the total cellular score grown when accompanying drawing display is and deducts do not add DHT with the Growth of Cells sum of 10nMDHT induction is the 100% allometry rejection ability carrying out computerized compound.