CN107311986A - The carboxylic acids inhibitor of heat shock protein 90 of Tetrahydroisoquinoli- beautiful jade 3 and its application - Google Patents
The carboxylic acids inhibitor of heat shock protein 90 of Tetrahydroisoquinoli- beautiful jade 3 and its application Download PDFInfo
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- CN107311986A CN107311986A CN201710551501.2A CN201710551501A CN107311986A CN 107311986 A CN107311986 A CN 107311986A CN 201710551501 A CN201710551501 A CN 201710551501A CN 107311986 A CN107311986 A CN 107311986A
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- heat shock
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/04—Ortho-condensed systems
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
- C07D495/04—Ortho-condensed systems
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The invention discloses a kind of carboxylic acids inhibitor of heat shock protein 90 of Tetrahydroisoquinoli- beautiful jade 3, the inhibitor of heat shock protein 90 is the compound with formula (I), or its optical isomer, diastereoisomer or its racemic mixture, or its pharmaceutically acceptable salt, solvate or prodrug.The invention also discloses application of the inhibitor in the medicine of prevention or treatment with heat shock protein 90 relevant disease is prepared.Experiment is confirmed：The carboxylic acids inhibitor of heat shock protein 90 of Tetrahydroisoquinoli- beautiful jade 3 of the present invention includes mammary gland to Various Tissues type, and the tumour cell that reproduction and respiratory system are originated has significant inhibitory action, but to Human normal hepatocyte small toxicity, with preferable druggability.Potential applicability in clinical practice is wide.
Description
Technical field
The present invention relates to a kind of inhibitor of heat shock protein 90 and its application, and in particular to a kind of Tetrahydroisoquinoli- beautiful jade -3- carboxylics
Acids inhibitor of heat shock protein 90 and its preparation method and application, belongs to organic compound synthesis and is led with medical applications technology
Domain.
Background technology
Heat shock protein 90 (Hsp90) is the highly conserved protein of a class formation during biological evolution.It is used as molecule
Companion, Hsp90 plays an important role in terms of regulation cell growth, differentiation, apoptosis.Research in recent years has shown that,
Hsp90 has close relationship with tumorigenesis, biological behaviour and its prognosis, and many oncogene proteins are Hsp90's
Action target spot.And Hsp90 inhibitor will promote the Hsp90 effect proteins played an important role in tumour growth signal path to drop
Solution, so as to block multiple target spots of tumor proliferation signal path, effectively prevents the growth of tumour.Inside and outside experiment is also confirmed that
The antitumor activity of Hsp90 inhibitor.The design of the micromolecular inhibitor based on dependency structure also causes increasingly extensive simultaneously
Concern.
Inventor is in the research of early stage, in order to find potent Hsp90 inhibitor that structure is novel, by design and rational and
Further compound structure modification is found that lead compound LCP1074 has stronger Hsp90 inhibitory activity (Chuan-
(2016) 272e282 of Peng Liang, European Journal of Medicinal Chemistry 121).Wherein
LCP1074 structural formulas are as follows：
Based on further structural modification is carried out to the compound, it is expected to obtain a class Hsp90 inhibitory activity and anti-thin
The compound that born of the same parents' proliferation activity all improves a lot.
The content of the invention
For the basis of prior art, the problem to be solved in the present invention is to provide a kind of Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids heat
Inhibitor of shock protein 90 and its preparation method and application.
Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90 of the present invention, it is characterised in that：The heat is stopped
Gram protein 90 inhibitor is the compound with formula (I), or its optical isomer, diastereoisomer or the mixing of its raceme
Thing, or its pharmaceutically acceptable salt, solvate or prodrug；
Wherein：
R1Selected from hydrogen-based, halogen, or C1-8 alkyl；
R2With R3It is each independently selected from hydrogen-based, deuterium base, or phosphonate group；
Ar is selected from the aromatic rings of substitution；Aromatic rings be selected from phenyl ring, five yuan of heteroaromatics, hexa-atomic heteroaromatic, it is hexa-atomic and five yuan it is miscellaneous
Ring, hexa-atomic and hexa-member heterocycle, five yuan and five-ring heterocycles, benzo five-membered heterocycle or various substituted benzo hexa-member heterocycles；Replace table
Show monosubstituted, disubstituted or three substitutions；Substituent is selected from hydrogen atom alkyl, halogen, hydroxyl, amino, dimethylamino, nitro, C1-
8 alkoxies, the miscellaneous alkoxies of C1-8, trifluoromethoxy, cyano group, carboxyl, sulfonic group or phosphate, or its ester or salt；
Phenyl ring A is optionally by hydrogen-based, one or more R4X group replaces；
X is selected from N, O, S atom or acyl group；
R4Selected from hydrogen-based, the cycloalkyl of C1-8 alkyl or 5-6 ring memberses, substituent is selected from C1-2 alkoxies, singly-or
Two-C1-2 alkyl aminos, the nitrogenous or oxygen-containing heterocycle of 5-6 ring memberses, hydroxyl, amino, nitro, cyano group, amide groups, first sulphur
Acylamino-, methyl sulfuryl, hydroxamic acid base, methoxycarbonyl group, hydrazide group, carboxyl, sulfonic group, phosphate, or its ester or salt；
* it is that spatial configuration is S or R optical purities or its raceme.
Further, above-mentioned inhibitor of heat shock protein 90 is preferably one of following compounds：
R1Selected from halogen；R2With R3It is hydrogen-based；7 of phenyl ring A are by R4X group replaces；X is selected from O atom；* it is three-dimensional structure
Type is R optical purities.
Most preferred embodiment is：The inhibitor of heat shock protein 90 is compound L 10, L15, L22 or L41, its
Chemical name order be：
R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -3- (N- is to cyanobenzyls-amide groups) -1,2,3,4- tetrahydrochysenes are different
Quinoline (L10)；
R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -3- (N- (pyridine -4- methyl)-amide groups) -1,2,3,4- tetrahydrochysenes
Isoquinolin (L15)；
R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -7- (2- methoxy ethoxies) -3- (N- (pyridine -4- methyl) -
Amide groups) -1,2,3,4- tetrahydroisoquinolines (L22)；
(N- is to cyanobenzyls-acid amides by -3- by R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -7- (2- morpholinoes ethyoxyl)
Base) -1,2,3,4- tetrahydroisoquinolines (L41).
The preparation method of above-mentioned Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90 be following synthetic route 1 or
Synthetic route 2, specific reactions steps and reaction equation are as follows：
Synthetic route 1：
With the Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids of 2,4- methyl dihydroxy benzoates, optical purity for initiation material.2,4- dihydroxies
Yl benzoic acid methyl esters is hydrolyzed after 3 chloros, benzyl protections, with Tetrahydroisoquinoli- beautiful jade -3- carboxylate methyl esters into acid amides, then through ester hydrolysis
Reacted afterwards with various amine, then debenzylation obtains target compound L1-20；Reaction equation is as follows：
Wherein, Ar is selected from the aromatic rings of substitution；Aromatic rings is selected from phenyl ring, five yuan of heteroaromatics, hexa-atomic heteroaromatic, hexa-atomic and five
Circle heterocycles, hexa-atomic and hexa-member heterocycle, five yuan and five-ring heterocycles, benzo five-membered heterocycle or various substituted benzo hexa-member heterocycles；Take
Monosubstituted, disubstituted or three substitutions are shown in representative；Substituent is selected from hydrogen atom alkyl, halogen, hydroxyl, amino, dimethylamino, nitre
Base, C1-8 alkoxies, the miscellaneous alkoxies of C1-8, trifluoromethoxy, cyano group, carboxyl, sulfonic group or phosphate, or its ester or
Salt；
Reagent in said synthesis route reaction equation：(a) sulfonic acid chloride, anhydrous methylene chloride；(b) benzyl bromine, potassium carbonate, N, N-
Dimethylformamide；(c) potassium hydroxide aqueous solution, methanol；(d) thionyl chloride, methanol；(e) I-hydroxybenzotriazole, 1- (3-
Dimethylamino-propyl) -3- ethyl-carbodiimide hydrochlorides, dichloromethane；(f) sodium hydrate aqueous solution, methanol；(g) various virtues
Cyclohexyl methyl amine, I-hydroxybenzotriazole, 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, N, N- dimethyl
Formamide；(h) boron chloride, anhydrous methylene chloride.
The structural formula of the target compound of synthetic route 1 is as follows：
Synthetic route 2：
With optically pure 3,5- diiodotyrosines for raw material, compound 10, chemical combination are obtained through Pictet-Spengler cyclizations
Thing 10 protects secondary amine, hydro-reduction to take off iodine and obtains key intermediate 12 through Boc in succession.Intermediate 12 and various fragrant methyl amines
The different Ar groups of connection are condensed through polypeptide to obtain compound 13.13 and take off Boc being condensed to yield corresponding chemical combination with intermediate 4 respectively
Thing 15.Compound 15 is prolonged with various halides or alcohol through Williamson's condensation reaction or light reacts into ether, then debenzylation obtains mesh
Mark compound L 21-45；Reaction equation is as follows：
Wherein, Ar is selected from the aromatic rings of substitution；Aromatic rings is selected from phenyl ring, five yuan of heteroaromatics, hexa-atomic heteroaromatic, hexa-atomic and five
Circle heterocycles, hexa-atomic and hexa-member heterocycle, five yuan and five-ring heterocycles, benzo five-membered heterocycle or various substituted benzo hexa-member heterocycles；Take
Monosubstituted, disubstituted or three substitutions are shown in representative；Substituent is selected from hydrogen atom alkyl, halogen, hydroxyl, amino, dimethylamino, nitre
Base, C1-8 alkoxies, the miscellaneous alkoxies of C1-8, trifluoromethoxy, cyano group, carboxyl, sulfonic group or phosphate, or its ester or
Salt；R4Selected from hydrogen-based, the cycloalkyl of C1-8 alkyl or 5-6 ring memberses, substituent is selected from C1-2 alkoxies, list-or two-C1-2
Alkyl amino, the nitrogenous or oxygen-containing heterocycle of 5-6 ring memberses, hydroxyl, amino, nitro, cyano group, amide groups, methanesulfonamido,
Methyl sulfuryl, hydroxamic acid base, methoxycarbonyl group, hydrazide group, carboxyl, sulfonic group, phosphate, or its ester or salt；
Reagent in said synthesis route reaction equation：(a) paraformaldehyde, concentrated hydrochloric acid, glycol dimethyl ether；(b)l mol/L
Sodium hydroxide solution, dimethyl dicarbonate butyl ester, tetrahydrofuran；(c) palladium carbon, hydrogen, triethylamine, methanol；(d) various aromatic ring methyl
Amine, I-hydroxybenzotriazole, 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, DMF；
(e) trifluoroacetic acid, triethylamine, dichloromethane；(f) intermediate 4, I-hydroxybenzotriazole, 1- (3- dimethylamino-propyls) -3- second
Base carbodiimide hydrochloride, N,N-dimethylformamide；(g) various halides, cesium carbonate, DMF；(h) three
Boron chloride, anhydrous methylene chloride.
The structural formula of the target compound of synthetic route 2 is as follows：
Concrete operation step prepared by above-claimed cpd will be described in detail in embodiment.
Those skilled in the art can be changed to improve yield to above-mentioned steps, and they can knowing according to this area substantially
Know the route for determining synthesis, such as selection reactant, solvent and temperature, can be avoided by using various GPF (General Protection False bases secondary anti-
The generation answered is so as to improve yield.
Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90 of the present invention is stopped in preparation prevention or treatment with heat
Application in the medicine of gram protein 90 relevant disease.
Wherein：The disease related to heat shock protein 90 be：Cancer, neurodegenerative disease, virus infection, inflammation,
Leukaemia, malaria or diabetes.
The invention also discloses a kind of prevention or treatment for being suitable to orally give mammal is related to heat shock protein 90
The pharmaceutical composition of disease, it is characterised in that：The pharmaceutical composition contain the Tetrahydroisoquinoli- beautiful jade of the present invention of therapeutically effective amount-
3- carboxylic acids inhibitor of heat shock protein 90 and one or more pharmaceutically acceptable carriers or excipient.
It is suitable to the parenteral prevention or treatment for giving mammal and heat shock protein 90 phase the invention also discloses a kind of
The pharmaceutical composition of related disorders, it is characterised in that：The pharmaceutical composition contains described in the claim 1,2 or 3 of therapeutically effective amount
Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90 and one or more pharmaceutically acceptable carriers or excipient.
Above-mentioned carrier can be salt solution, buffered saline, glucose, water, glycerine, ethanol or their conjugate；Either
Solid carrier lactose, land plaster, sucrose, talcum, gel, agar, pectin, Arabic gum, magnesium stearate or stearic acid etc..
The present invention is based on carrying out compound L CP1074 further structural modification, and screening obtains class Hsp90 suppression
There is provided a kind of Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids heat shock proteins for the compound that activity and antiproliferation are significantly increased
White 90 inhibitor, and suitable for orally give mammal suitable for the parenteral prevention for giving mammal or treatment with heat
The pharmaceutical composition of the relevant disease of shock protein 90.Experiment is confirmed：Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids the heat shock proteins of the present invention
90 inhibitor include mammary gland to Various Tissues type, and there is the tumour cell that reproduction and respiratory system are originated significant suppression to make
With, but to Human normal hepatocyte small toxicity, with preferable druggability.Potential applicability in clinical practice is wide.
Brief description of the drawings
Fig. 1：The intracellular of compound L 15 suppresses Hsp90 activity ratings.
Embodiment
To enable this area related researcher's comprehensive understanding present invention, illustrate the present invention's with reference to embodiment
Several embodiments, although associated description is more specific and detailed, therefore can not limit the present invention.
Embodiment 1：Compound L 1-20 synthesis, by taking L1 as an example.
1) the chloro- 2,4- methyl dihydroxy benzoates 2 of 5-
It is that 2,4- methyl dihydroxy benzoates (8.0g, 47.6mmol) are dissolved in anhydrous methylene chloride by compound 1, cools down
After to 0 DEG C add sulfonic acid chloride (4.0mL, 49.4mmol), be stirred at room temperature after 13 hours, again add sulfonic acid chloride (2mL,
24.7mmol), saturation NaHCO3 (7.5mL) quenching reactions are added after stirring 6 hours.Organic phase, aqueous phase dichloro are collected in extraction
Methane is extracted three times, is merged silica gel column chromatography after organic phase, anhydrous sodium sulfate drying, solvent evaporated and is purified to obtain white solid chemical combination
The common 5.78g of thing 2, yield：60%.1H NMR(600MHz,CDCl3,r.t.)δ10.82(s,1H),7.82(s,1H),6.61(s,
1H),5.94(s,1H),3.93(s,3H).
2) the chloro- methyl benzoates 3 of 2,4- benzyloxies -5-
Compound 2 (5.78g, 28.56mmol) and potassium carbonate (8.68g, 62.83mmol) are dissolved in 50mL DMF, stirred
8.20mL benzyl bromines are added after 5 minutes.React 6 hours, be cooled to after room temperature at 60 DEG C, addition water and ethyl acetate, extracting and demixing,
Organic phase is washed with saturated common salt, anhydrous sodium sulfate drying, and silica gel column chromatography purifies to obtain 9.43g white solids 3 after solvent evaporated.
Yield:86.2%.1H NMR(600MHz,CDCl3,r.t.)δ7.94(s,1H),7.44-7.31(m,10H),6.56(s,1H),
5.12(s,2H),5.10(s,2H),3.87(s,3H).HR ESIMS:m/z 405.0869[M+Na]+
(calcd.405.0870).
3) the chloro- benzoic acid 4 of 2,4- benzyloxies -5-
Compound 3 (9.43g, 24.62mmol) is added to after 50mL methanol, 20mL potassium hydroxide (7.06g) is added
The aqueous solution, return stirring 24 hours.Reaction solution is cooled to after room temperature, after hydrochloric acid solution to the acidity for adding 5%, ethyl acetate extraction
Take, organic phase saturated common salt water process, anhydrous sodium sulfate drying, solvent evaporated obtains the common 8.8g of white solid 4, yield：97%
。1H NMR(600MHz,CDCl3,r.t.)δ8.19(s,1H),7.41-7.36(m,10H),6.61(s,1H),5.19(s,2H),
5.18(s,2H).HR ESIMS:m/z 367.0739[M-H]-(calcd.367.0737).
4) 1,2,3,4- tetrahydroisoquinoline -3- carboxylate methyl ester hydrochlorides 6
Compound 5 (4.27g, 20mmol) is added in 20mL methanol, instillation 5.2mL thionyl chlorides, room after 0 DEG C is cooled to
After temperature stirring 100 hours, decompression is spin-dried for, and residue is washed with ether, and 4.3g faint yellow solids compound 6 is obtained after drying, and is received
Rate：94.4%.1H NMR(300MHz,D2O, r.t.) δ 7.45-7.24 (m, 4H), 4.60 (d, J=5.5Hz, 1H), 4.56 (d, J
=5.4Hz, 1H), 4.53 (br s, 1H), 3.93 (s, 3H), 3.53 (dd, J=17.4,5.5Hz, 1H), 3.53 (dd, J=
17.4,5.5Hz, 1H), 3.32 (dd, J=17.3,10.9Hz, 1H), 3.32 (dd, J=17.3,10.9Hz, 1H) .HR
ESIMS:m/z192.1028[M+H]+(calcd.192.1025).
5) 2- (2, the 4- chloro- benzoyls of benzyloxy -5-) -1,2,3,4- tetrahydroisoquinoline -3- carboxylate methyl esters 7
Compound 4 (2.213g, 6mmol) is dissolved in 30mL anhydrous methylene chlorides with compound 6 (1.639g, 7.2mmol),
Sequentially add I-hydroxybenzotriazole (HOBt, 0.973g, 7.2mmol), 1- (3- dimethylamino-propyls) -3- ethyl carbodiimides
Hydrochloride (EDCI, 1.38g, 7.2mmol), N-methylmorpholine (NMM, 2.012mL, 18mmol), are stirred at room temperature 12 hours.Reaction
Liquid successively use water, 5% hydrochloric acid solution, saturation NaHCO3, saturated common salt water process, separate organic phase, anhydrous sodium sulfate drying,
Silica gel column chromatography purifies to obtain 2.89g compounds 7, yield after solvent evaporated：81%.1H NMR(300MHz,CDCl3,r.t)δ
7.52–7.05(m,16H),5.30–5.12(m,4H),4.41(s,1H),3.59–3.45(m,2H),3.30(s,3H),3.14–
3.00(m,2H).HR ESIMS:m/z 542.1731[M+H]+(calcd.542.1734).
6) R-2- (2, the 4- chloro- benzoyls of benzyloxy -5-) -1,2,3,4- tetrahydroisoquinoline -3- carboxylic acids 8
Compound 7 (2.89g, 5.338mmol) is dissolved in 16mL methanol, 0.62mL2mol/L sodium hydroxide is added
The aqueous solution.12h is stirred at room temperature.Acid liquid adjusts pH value to weak acid after completion of the reaction, and ethyl acetate is extracted, after anhydrous sodium sulfate drying
It is evaporated to obtain 2.68g compounds 8, yield：95%.1H NMR(300MHz,CDCl3,r.t)δ12.85(s,1H),7.52–7.05(m,
16H),5.30–5.12(m,4H),4.41(s,1H),3.59–3.45(m,2H),3.14–3.00(m,2H).HR ESIMS:m/
z526.1418[M-H]-(calcd.526.1421).
7) R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -3- (N- p-chlorobenzyls-amide groups) -1,2,3,4- tetrahydrochysenes are different
Quinoline L1
Compound 8 (100mg, 0.19mmol), I-hydroxybenzotriazole (HOBt, 30.8mg, 0.228mmol), 1- (3- bis-
Methylaminopropyl) -3- ethyl-carbodiimide hydrochlorides (EDCI, 43.7mg, 0.228mmol) are dissolved in 3mL DMF, stir 3 points
Added after clock 115 μ L to chlorobenzylamine, be stirred at room temperature 12 hours.Reaction solution is molten with water, saturation NaHCO3,5% hydrochloric acid successively
Liquid, saturated common salt water process, separate organic phase, anhydrous sodium sulfate drying, solvent evaporated, the not purified directly use of gained compound
In next step.Gained compound is dissolved in anhydrous methylene chloride, addition 0.57mL boron chlorides after 0 DEG C is cooled under N2 protections molten
It is stirred at room temperature after liquid (3eq), 20min 2 hours, saturated sodium bicarbonate solution is added under ice bath to weak base, quenching reaction removes two
Chloromethanes layer, ethyl acetate aqueous phase extracted, salt washing, anhydrous Na 2SO4 is dried, and silica gel column chromatography is purified after solvent evaporated
81.3mg white solid A1. yields：76%.1H NMR(400MHz,Acetone)δ7.70(br s,1H),7.36(s,1H),
7.27–7.20(m,3H),7.19(br s,1H),7.10(s,1H),7.09(s,1H),7.04-7.00(m,1H),6.88(br
S, 1H), 6.61 (s, 1H), 5.14 (br s, 1H), 4.80-4.70 (m, 2H), 4.37 (dd, J=15.2,5.7Hz, 1H), 4.28
(dd, J=15.3,5.3Hz, 1H), 3.37 (dd, J=15.4,4.9Hz, 1H), 3.27 (dd, J=15.5,6.0Hz, 1H) .HR
ESIMS:m/z for 471.0846[M+H]+,(calcd.471.0878)。
Embodiment 2：Compound L 21-45 synthesis, by taking L21 as an example.
1) the iodo- 1,2,3,4- tetrahydroisoquinolines -3- carboxylic acid hydrochlorides 10 of R-7- hydroxyls -6,8- two
The iodo- D-Tyrosines (1.0g, 2.31mmol) of 3,5- bis-, glycol dimethyl ether (0.7mL) are added into 8mL concentrated hydrochloric acids
With paraformaldehyde (0.26g, 8.67mmol), and 72 DEG C are gradually heating to.After 0.5 hour, concentrated hydrochloric acid (1.7mL), second are added
Glycol dimethyl ether (0.33mL) and poly formic acid (0.17g, 5.78mmol), oil bath temperature control continue to react 18 hours at 72-75 DEG C.
Reaction suspension ice bath is cooled down and filtered, and filter cake spent glycol dimethyl ether fully washes paint, and 0.53g pale yellow powders are obtained after drying.1H NMR(300MHz,DMSO-d6, r.t) and δ 3.07 (dd, J=16.8Hz, 10.8Hz, 1H), 3.22 (dd, J=16.8Hz,
4.8Hz, 1H), 4.02 (d, J=16.2Hz, 1H), 4.15 (d, J=16.2Hz, 1H), 4.32 (dd, J=4.8Hz, 10.8Hz,
1H),7.73(s,1H),9.68(s,1H),10.00(br s,2H),14.17(br s,1H),ESI-MS m/z:446.2[M+H
]+
2) the iodo- 1,2,3,4- tetrahydroisoquinolines -3- carboxylic acid hydrochlorides 11 of R-2- tertbutyloxycarbonyls 7- hydroxyls -6,8- two
Compound 10 (0.53g) is dissolved in 2.5mLlmol/L oxygen sodium hydroxide solution, and adds 0.6mL dimethyl dicarbonate fourths
The tetrahydrofuran solution of ester (0.27g).Reaction solution pH is controlled in 9-11 with 1mol/L sodium hydroxide solution in course of reaction.Room
After temperature reaction 6 hours, the tetrahydrofuran in reaction solution is evaporated off, then extracted reaction solution 3 times with petroleum acids, and with 1mol/L lemon
Lemon acid solution is acidified to pH4-5, is then extracted with ethyl acetate three times, and organic phase uses saturated common salt water washing after merging, anhydrous
Sodium sulphate is dried, and solvent evaporated obtains 0.8g yellow powders 11.Yield:92%,1H NMR(300MHz,DMSO-d6,r.t)δ1.34+
1.40(s,9H),2.87-3.00(m,2H),4.13-4.41(m,2H),4.61-4.75(m,1H),7.57(s,1H),9.41(br
s,1H),12.71(br s,1H)。
3) the oxygen isoquinolin -3- carboxylic acid hydrochlorides 12 of R-2- tertbutyloxycarbonyls 7- hydroxyls -1,2,3,4- four
Compound 11 (0.8g) is dissolved in 10mL absolute methanols, and adds triethylamine (480 μ L) and 10% palladium carbon thereto
(0.08g).After being passed through hydrogen reaction 5 hours, filtered off through Celite catalyst is used, methanol is evaporated off, the citric acid for adding 1mol/L is molten
Then liquid be extracted with ethyl acetate three times to pH4-5, is washed and painted with saturated common salt after combined ethyl acetate layer, anhydrous sodium sulfate
Dry, solvent evaporated obtains 0.36g yellow powders 12.Yield:75%,1H NMR(300MHz,DMSO-d6,r.t)δ1.39+1.45
(s, 9H), 2.92-3.04 (m, 2H), 4.26-4.51 (m, 2H), 4.57-4.82 (m, 1H), 6.52 (s, 1H), 6.57 (d, J=
8.4Hz, 1H), 6.97 (d, J=8.4Hz, 1H), 9.28 (s, 1H), 12.60 (s, 1H).
4) R-2- tertbutyloxycarbonyls -7- hydroxyls -3- (N- (pyridine -4- methyl)-amide groups) -1,2,3,4- tetrahydroisoquinolines
13
Compound 12 (0.36g), 4- methylamino pyridines (500 μ L) are dissolved in 10mL DMF, sequentially add 1- hydroxy benzos
Triazole (HOBt, 0.2g), 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides (EDCI, 0.28g), are stirred at room temperature
12 hours.Reaction solution uses water, saturation NaHCO successively3, 10% stubborn lemon acid solution, saturated common salt water process, separate organic phase,
The white solid 0.26g. yields of silica gel column chromatography after anhydrous sodium sulfate drying, solvent evaporated:75%,1H NMR(300MHz,
Acetone-d6,r.t)δ1.39+1.45(s,9H),2.92-3.04(m,2H),3.57-3.82(m,2H),4.26-4.51(m,
2H), 4.57-4.82 (m, 1H), 6.72-6.80 (m, 4H), 7.02 (d, J=8.1Hz, 1H), 7.68 (s, 1H), 8.31-8.37
(m,2H).
5) R-2- (the chloro- benzoyls of 2,4- benzyloxies -5-) -7- hydroxyls -3- (N- (pyridine -4- methyl)-amide groups) -
1,2,3,4- tetrahydroisoquinoline 14
Burnt to the 3mL dichloromethanes of compound 13 (410mg) and 1mL trifluoroacetic acids are added in solution, reaction terminates backward reaction
Excess of triethylamine is added in liquid stand-by to alkalescence.Into the anhydrous tetra oxygen furyl solution of 3mL dissolved with intermediate 4 (413mg, 1eq),
Add triethylamine, I-hydroxybenzotriazole (HOBt, 182mg), 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides
(EDCI258mg), it is stirred at room temperature after 15 minutes, this reaction solution is poured into above-mentioned dichloromethane institute solution.It is evaporated off after reaction overnight
Solvent, adds ethyl acetate, successively with water, saturation NaHCO3, saturated common salt water process, separate organic phase, anhydrous sodium sulfate do
It is dry, the white solid 600mg. of silica gel column chromatography after solvent evaporated
7) R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -7- methoxyl groups -3- (N- p-chlorobenzyls-amide groups) -1,2,3,
4- tetrahydroisoquinolines L21
Cesium carbonate powder (150mg) and N- (2- chloroethyls) morpholine is added in the 4mLDMF solution of compound 14 (50mg)
Hydrochloride (37mg), ethyl acetate is extracted three times after being stirred overnight at room temperature, and is washed and painted with saturated common salt after combined ethyl acetate layer,
Anhydrous magnesium sulfate is dried, and solvent evaporated obtains product crude product.Gained compound is not purified to be directly used in next step.Gained compound
It is dissolved in anhydrous methylene chloride, N2It is cooled under protection after 0 DEG C and is stirred at room temperature 2 after addition 0.57mL boron chloride solution, 20min
Hour, saturated sodium bicarbonate solution is added under ice bath to weak base, dichloromethane layer, ethyl acetate extraction water are removed in quenching reaction
Phase, salt washing, anhydrous Na2SO4Dry, silica gel column chromatography purifies to obtain 81.3mg white solid A1. yields after solvent evaporated：76%
。1H NMR(400MHz,Acetone-d6,r.t)δ7.70(br s,1H),7.36(s,1H),7.27–7.20(m,3H),7.19
(br s,1H),7.10(s,1H),7.09(s,1H),7.04-7.00(m,1H),6.61(s,1H),5.14(br s,1H),
4.80-4.70 (m, 2H), 4.37 (dd, J=15.2,5.7Hz, 1H), 4.28 (dd, J=15.3,5.3Hz, 1H), 3.76 (s,
3H), 3.37 (dd, J=15.4,4.9Hz, 1H), 3.27 (dd, J=15.5,6.0Hz, 1H).
Embodiment 3：Tetrahydroisoquinoli- beautiful jade -3- carboxylic acid derivative extracorporeal anti-tumor cytoactive screening tests
Method of testing：
The antitumor activity of various compounds is evaluated with srb assay.The cell in exponential phase is taken, with
2000-8000cells/well density is inoculated in 96 orifice plates, dosing (compound L 1-L45) after 12h cell attachments, is continued
Cultivate 72h.Cell culture terminates rear every hole plus the trichloroacetic acids of 100 μ L 10% (TCA) are fixed, stands 5min and goes to 4 DEG C of placements
1h.Cell is fixed and finished, and abandons TCA, with distillation washing 5 times, dries and static 5min is dried in atmosphere.50 μ L are added per hole
Under 4mg/mLSRB solution, room temperature condition stand dyeing 10 minutes, washed away with 1% glacial acetic acid not with protein bound SRB, wash 5 times
Dry afterwards and be placed in air drying.With protein bound SRB, 200 μ L 10mM Tris non-buffered solution are added per hole, hand is used
1min is patted fully to dissolve.Spectrophotometric determination OD values are finally used, measure wavelength is 570nm.
The inhibiting rate of tumor cell proliferation is calculated as follows：
IC50Represent sample to the concentration when inhibiting rate of tumour cell is 50%.
Test material：
Human breast cancer cell MDA-MB-231, MDA-MB-453；Human neuroblastoma cells SHSY5Y；People's glioma is thin
Born of the same parents U251；Human colon cancer cell HCT116, SW480；Human lung carcinoma cell H1299, A549；Human cervical carcinoma cell HeLa, people prostatitis
Gland cancer PC3；Human liver cancer cell HepG2；Human normal hepatocyte HL7702.It is purchased from ATCC.
Test result：It is shown in Table 1.
Table 1：Activity of the part Tetrahydroisoquinoli- beautiful jade -3- carboxylic acid derivatives to 12 plants of cells
In summary, compound L 10, L15, L22 and L41 include mammary gland, reproduction and respiratory system to Various Tissues type
The tumour cell in source has significant inhibitory action, but to Human normal hepatocyte small toxicity, with preferable druggability.
Embodiment 4：Hsp90 inhibitory activity is evaluated
Representative compound L15 is chosen, its influence in cellular level to Hsp90 GAP-associated protein GAPs is detected.Hsp90 client's egg
The lower mediation Hsp70 of white level covering up-regulation is two most common Biological indicators for weighing compound suppression Hsp90.
Westernblot results show that the intracellular of compound L 15 acts on Hsp90.
Method of testing：
Take the MDA-MB231 cells in logarithmic growth to be inoculated in Tissue Culture Dish, treat cell growth to 70% density
When, culture medium is abandoned in agent-feeding treatment corresponding time, suction, washed once with the PBS of precooling on ice, adds cell pyrolysis liquid processing.It is molten
Solve product and add isometric 2 × SDS sample buffers, 5min is boiled in heating makes albumen thoroughly be denatured.The sample of denaturation is passed through first
8%-12%SDS-PAGE is crossed, then electricity is gone on polyvinylidene fluoride (PVDF) film of methanol activation.After electrotransfer
Pvdf membrane is placed in confining liquid (5%BSA), decolorization swinging table room temperature closing 1h.Then, add and diluted with 5%BSA by corresponding proportion
Primary antibody solution, 4 DEG C of closings are stayed overnight, and TBST is washed 3 times, each 10min.Add the two of corresponding coupling horseradish peroxidase
Anti- solution, is placed in decolorization swinging table and shakes 1h, TBST is washed 3 times, each 10min.Pvdf membrane enhancedization newly matched somebody with somebody after washing
Luminous agent solution impregnation is learned, observation of taking pictures is placed in ChemDoc XRS+ gel imagers.
Test result：See Fig. 1.
Claims (7)
1. a kind of Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90, it is characterised in that：The heat shock protein 90 suppression
Preparation is the compound with formula (I), or its optical isomer, diastereoisomer or its racemic mixture, or its medicine
Acceptable salt, solvate or prodrug on；
Wherein：
R1Selected from hydrogen-based, halogen, or C1-8 alkyl；
R2With R3It is each independently selected from hydrogen-based, deuterium base, or phosphonate group；
Ar is selected from the aromatic rings of substitution；Aromatic rings is selected from phenyl ring, five yuan of heteroaromatics, hexa-atomic heteroaromatic, hexa-atomic and five-ring heterocycles, six
Member and hexa-member heterocycle, five yuan and five-ring heterocycles, benzo five-membered heterocycle or various substituted benzo hexa-member heterocycles；Substitution represents singly to take
Generation, disubstituted or three substitutions；Substituent is selected from hydrogen atom alkyl, halogen, hydroxyl, amino, dimethylamino, nitro, C1-8 alcoxyls
Base, the miscellaneous alkoxies of C1-8, trifluoromethoxy, cyano group, carboxyl, sulfonic group or phosphate, or its ester or salt；
Phenyl ring A is optionally by hydrogen-based, one or more R4X group replaces；
X is selected from N, O, S atom or acyl group；
R4Selected from hydrogen-based, the cycloalkyl of C1-8 alkyl or 5-6 ring memberses, substituent is selected from C1-2 alkoxies, list-or two-C1-2
Alkyl amino, the nitrogenous or oxygen-containing heterocycle of 5-6 ring memberses, hydroxyl, amino, nitro, cyano group, amide groups, methanesulfonamido,
Methyl sulfuryl, hydroxamic acid base, methoxycarbonyl group, hydrazide group, carboxyl, sulfonic group, phosphate, or its ester or salt；
* it is that spatial configuration is S or R optical purities or its raceme.
2. Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90 as claimed in claim 1, it is characterised in that：It is described
Inhibitor of heat shock protein 90 is one of following compounds：
R1Selected from halogen；R2With R3It is hydrogen-based；7 of phenyl ring A are by R4X group replaces；X is selected from O atom；* it is that spatial configuration is R
Optical purity.
3. Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90 as claimed in claim 2, it is characterised in that：It is described
Inhibitor of heat shock protein 90 is compound L 10, L15, L22 or L41, and its chemical name order is：
R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -3- (N- is to cyanobenzyls-amide groups) -1,2,3,4- tetrahydroisoquinolines
(L10)；
R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -3- (N- (pyridine -4- methyl)-amide groups) -1,2,3,4- Tetrahydroisoquinoli-s
Quinoline (L15)；
R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -7- (2- methoxy ethoxies) -3- (N- (pyridine -4- methyl)-acid amides
Base) -1,2,3,4- tetrahydroisoquinolines (L22)；
R-2- (chloro- 2, the 4- dihydroxybenzoyls of 5-) -7- (2- morpholinoes ethyoxyl) -3- (N- is to cyanobenzyls-amide groups) -
1,2,3,4- tetrahydroisoquinoline (L41).
4. Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids inhibitor of heat shock protein 90 described in claim 1,2 or 3 is preparing prevention or treated
With the application in the medicine of heat shock protein 90 relevant disease.
5. application as claimed in claim 4, it is characterised in that：The disease related to heat shock protein 90 be：Cancer, god
Through degenerative disease, virus infection, inflammation, leukaemia, malaria or diabetes.
6. a kind of pharmaceutical composition for being suitable to orally give the prevention or treatment and heat shock protein 90 relevant disease of mammal,
It is characterized in that：The pharmaceutical composition contains Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids described in the claim 1,2 or 3 of therapeutically effective amount
Inhibitor of heat shock protein 90 and one or more pharmaceutically acceptable carriers or excipient.
7. a kind of drug regimen suitable for the parenteral prevention or treatment and heat shock protein 90 relevant disease for giving mammal
Thing, it is characterised in that：The pharmaceutical composition contains Tetrahydroisoquinoli- beautiful jade -3- carboxylic acids described in the claim 1,2 or 3 of therapeutically effective amount
Class inhibitor of heat shock protein 90 and one or more pharmaceutically acceptable carriers or excipient.
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|---|---|---|---|---|
| CN101687808A (en) * | 2007-06-21 | 2010-03-31 | 默克专利有限公司 | indazolamide derivatives |
| CN103724269A (en) * | 2012-10-11 | 2014-04-16 | 中国科学院上海药物研究所 | Phenyl 1,2-isoxazole or phenyl 1,2-pyrazole compounds and application thereof |
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2017
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101687808A (en) * | 2007-06-21 | 2010-03-31 | 默克专利有限公司 | indazolamide derivatives |
| CN103724269A (en) * | 2012-10-11 | 2014-04-16 | 中国科学院上海药物研究所 | Phenyl 1,2-isoxazole or phenyl 1,2-pyrazole compounds and application thereof |
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| CHUANPENG LIANG ET AL.: "Design and synthesis of N-(5-chloro-2,4-dihydroxybenzoyl)-(R)-1,2,3,4-tetrahydroisoquinoline-3-carboxamides as novel Hsp90 inhibitors", 《EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY》 * |
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| CN115124550A (en) * | 2020-10-26 | 2022-09-30 | 山东大学 | Preparation and application of specific heat shock protein 90 alpha subtype inhibitor |
| CN115124550B (en) * | 2020-10-26 | 2024-04-02 | 山东大学 | Preparation and application of specific heat shock protein 90 alpha subtype inhibitor |
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