CN116217457A - 4-amino-1H-pyrrole compound, preparation method and application - Google Patents
4-amino-1H-pyrrole compound, preparation method and application Download PDFInfo
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Abstract
The invention discloses a 4-amino-1H-pyrrole compound, a preparation method and application thereof. The invention provides a series of 4-amino-1H-pyrrole compounds, an activity test shows that the compounds have Mcl-1 inhibition effect, most of the compounds have nanomolar Mcl-1 inhibition activity, have good selectivity on other subtypes Bcl-2, bcl-xL, bcl-w and Bfl-1 of Bcl-2 anti-apoptosis protein family, and also show good activity in-vitro anti-tumor proliferation activity evaluation, wherein the compound 52 with the best activity has good drug-like quality, and shows good anti-tumor activity in a Mv4-11 xenograft tumor nude mouse model. Therefore, the 4-amino-1H-pyrrole compound provided by the invention has the prospect of being developed into an Mcl-1 inhibitor drug, and is hopeful to be developed into an antitumor drug.
Description
Technical Field
The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a 4-amino-1H-pyrrole compound, a preparation method and application thereof.
Background
Apoptosis, also known as apoptosis, plays an important role in the clearance of harmful and potentially dangerous cells and in maintaining the intracellular environment. Mcl-1 is often overexpressed in tumor cells as an anti-apoptotic protein and is closely related to the occurrence of cancer and the development of drug resistance, and is thus being studied intensively as a potential target for tumor treatment.
An important process in the endogenous apoptotic pathway is that the pro-apoptotic proteins Bax/Bak bind to each other to form dimers that depolarize the mitochondrial outer membrane, and apoptosis factors such as cytochrome c outflow induce downstream signal transduction. Mcl-1 is able to inhibit apoptosis by inhibiting its dimer formation through its hydrophobic groove binding to Bax/Bak.
Small molecule BH3 mimics that mimic the binding of the BH3 domain to Mcl-1 proteins, thereby inhibiting the interaction of Mcl-1 with pro-apoptotic proteins, a strategy that is widely used in the design of Mcl-1 small molecule inhibitors. In recent years, more and more small molecule inhibitors have been reported, and some Mcl-1 with high activity and good drug formation is being clinically studied, and is expected to become a new cancer therapeutic drug (Troae et al, nat Commun.9 (2018): 5341).
Disclosure of Invention
The invention aims to provide a 4-amino-1H-pyrrole compound, a preparation method and application.
The above object of the present invention is achieved by the following technical scheme:
A4-amino-1H-pyrrole compound has the following structural general formula:
R 1 and R is 2 Selected from the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, halogen, hydroxy, mercapto, alkoxy, and alkylthio; r is R 3 And R is 4 Each independently selected from a hydrogen atom, a substituted or unsubstituted C1-C5 alkyl group.
Preferably, the alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl substituted with one or more substituents selected from the group consisting of alkyl, halogen, hydroxy, mercapto, amino, oxo, carboxyl, nitro, cyano, alkoxy, alkylthio, cycloalkyl, heterocyclyl, aryl and heteroaryl.
More preferably, R 1 Selected from phenyl, 2-pyridyl, 1-naphthyl, 2-methylphenyl, 3-methylphenyl, 4-ethylphenyl, 4-cyclopropylphenyl, 4-isopropylphenyl, 4-t-butylphenyl, 3-ethylphenyl, 3-isopropylphenyl, 3-t-butylphenyl, 3-chlorophenyl, 3-fluorophenyl, 3-acetylphenyl, 3-nitrophenyl, 3-cyanophenyl, 3-methoxyphenyl, 3-hydroxyphenyl, 3- (1-hydroxyethyl) phenyl, 3, 4-dimethylphenyl and 3-methyl-4-chlorophenyl.
More preferably, R 2 Selected from 3-thienyl, 1-naphthyl, 2-naphthyl, phenyl, 2-chlorophenyl, 2-methylphenyl, 3-chloromethyl, 3-methylphenyl, 4-chlorophenyl, 4-methylphenyl, 3-bromophenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-trifluoromethylphenyl, 3-trifluoromethoxyphenyl, 3-aldehydylphenyl, 4-isopropylphenyl, 4-tert-butylphenyl, 4-biphenylyl, 1-phenyl-1H-1, 2, 3-triazol-4-yl, 4-ethynylphenyl, 4-propynylphenyl, 3,4 dimethylphenyl, 3-bromo-4-methylphenyl, 3, 4-dichlorophenyl, 3,5 dimethylphenyl, 2, 3-difluorophenyl, 2, 4-dichlorophenyl, 2, 6-dichlorophenyl, 3-chloro-4 phenylphenyl and 3-chloro-4-ethynylphenyl.
Method for preparing 4-amino-1H-pyrrole compound, R 1 Definition of R is as defined in claim 1 2 =3-chlorophenyl group, R 3 、R 4 Is a hydrogen atom, and comprises the following synthetic routes and steps:
raw materials I-2 and 1, 3-dibromopropane are in acetonitrile, potassium carbonate is taken as an acid binding agent, nucleophilic substitution reaction is carried out to obtain an intermediate I-3, the intermediate I-1 and the intermediate I-3 are under the action of cesium carbonate to obtain an intermediate I-4 in DMF, the intermediate I-4 is subjected to nucleophilic substitution reaction to obtain I-5, I-5-and different bromoband aromatic hydrocarbons in Pd through stannous chloride reduction 2 (dba) 3 Under the catalysis of BINAP, cs is used 2 CO 3 As alkali, obtaining an intermediate I-6, I-6 and 3-chlorobenzyl through Buchward-Hartwig reaction, taking NaH as an acid binding agent, carrying out nucleophilic substitution reaction with 3-chlorobenzyl to obtain an intermediate I-7, then hydrolyzing the intermediate I-7 to obtain a target product and obtaining an intermediate I-8,I-8 through H 2 And (3) debenzylating Pt/C to obtain a target product a, and reducing the target product substituted by the 3-acetylphenyl R1 by sodium borohydride to obtain a target product b.
Method for preparing 4-amino-1H-pyrrole compound, R 1 =3-methylphenyl, R 2 Defined as follows, R 3 、R 4 Is a hydrogen atom, and comprises the following synthetic routes and steps:
R 2 3-thienyl, 1-naphthyl, 2-naphthyl, phenyl, 2-chlorophenyl, 2-methylphenyl, 3-chloromethyl, 3-methylphenyl, 4-chlorophenyl, 4-methylphenyl, 3-bromophenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-trifluoromethylphenyl, 3-trifluoromethoxyphenyl, 3-aldehydylphenyl, 4-isopropylphenyl, 4-tert-butylphenyl, 4-biphenylyl, 1-phenyl-1H-1, 2, 3-triazol-4-yl, 4-ethynylphenyl, 4-propynylphenyl, 3,4 dimethylphenyl, 3-bromo-4-methylphenyl, 3, 4-dichlorophenyl, 3,5 dimethylphenyl, 2, 3-difluorophenyl, 2, 4-dichlorophenyl or 2, 6-dichlorophenyl;
NaH is used as an acid binding agent, intermediate I-9 and bromobenzyl with different substitutions are subjected to nucleophilic substitution reaction to obtain intermediate II-1, II-2, the intermediate II-1 and II-2 are hydrolyzed by NaOH to obtain a target product a, one of the intermediate II-2 and azidobenzene are subjected to click reaction under the action of DIPEA and CuI to obtain intermediate II-3, and the intermediate II-3 is hydrolyzed by NaOH to obtain a target product b.
Method for preparing 4-amino-1H-pyrrole compound, R 1 3-methylphenyl, 3-chloromethyl, 3-fluoromethylphenyl, 3-methoxyphenyl, 3, 4-dimethylphenyl, 3-chloro-4-methylphenyl, R 2 =3-chloro-4-phenylphenyl, 3-chloro-4-ethynylphenyl, R 3 、R 4 Is hydrogen atom or methyl, comprising the following synthetic route and steps:
starting material III-1 with phenylboronic acid in Pd (PPh 3 ) 4 Under the catalysis, obtaining an intermediate III-2 through Suzuki reaction, obtaining benzyl alcohol III-3 through borane reduction of III-2, and obtaining bromobenzyl III-4 through reaction of III-3 and phosphorus tribromide; reducing III-1 with borane to obtain benzyl alcohol III-5, III-5 and trimethylsilylacetylene in PdCl 2 (PPh 3 ) 2 Under the catalysis of cuprous iodide, introducing alkynyl through Sonogashira coupling to obtain intermediate III-6, deprotecting III-6 with TBAF to obtain III-7, and mixing with PBr 3 Obtaining bromobenzyl III-8 by reaction, obtaining intermediate III-10 by nucleophilic substitution reaction of intermediate III-9 and III-4 and III-8 under the action of NaH, and obtaining target product by NaOH hydrolysis of intermediate III-10.
Use of the above 4-amino-1H-pyrrole compound for the preparation of a Mcl-1 inhibitor medicament.
The application of the 4-amino-1H-pyrrole compound in preparing antitumor drugs.
The beneficial effects are that:
the invention provides a series of 4-amino-1H-pyrrole compounds, an activity test shows that the compounds have Mcl-1 inhibition effect, most of the compounds have nanomolar Mcl-1 inhibition activity, have good selectivity on other subtypes Bcl-2, bcl-xL, bcl-w and Bfl-1 of Bcl-2 anti-apoptosis protein family, and also show good activity in-vitro anti-tumor proliferation activity evaluation, wherein the compound 52 with the best activity has good drug-like quality, and shows good anti-tumor activity in a Mv4-11 xenograft tumor nude mouse model. Therefore, the 4-amino-1H-pyrrole compound provided by the invention has the prospect of being developed into an Mcl-1 inhibitor drug, and is hopeful to be developed into an antitumor drug.
Drawings
FIG. 1 is a graph of affinity evaluation of compound 52 for Bcl-2 subtype;
FIG. 2 shows the results of in vivo antitumor activity studies of compound 52; wherein: a is the change in tumor volume during administration; the long diameter a and the short diameter b of the tumor are measured during the administration period, and the tumor volume is calculated according to the formula v=a×b 2 Calculation of p<0.01，****p<0.0001 (n=6), one-wayANOVAwith Tukey-Kramer posttest; b is the trend of change in body weight of nude mice during administration; c is the weight of each group of heart, liver and kidney after administration; d is tumor tissue isolated after the end of administration.
Detailed Description
The following describes the essential aspects of the present invention in detail with reference to examples, but is not intended to limit the scope of the present invention.
Example 1:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (phenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 1)
(1) Preparation of 5- (3-bromopropyloxy) -2-chloro-1, 3-dimethylbenzene
4-chloro-3, 5-dimethylphenol (25.6 mmol,4g,1 eq) was dissolved in 40ml acetonitrile, followed by addition of potassium carbonate (3 eq) and 1, 3-dibromopropane (3 eq) and reflux for 3h. After the reaction was completed, it was cooled to room temperature, 150ml of water was added, EA was extracted 3 times, the organic phase was combined, dried with saturated brine, dried over sodium sulfate, suspended, and sand-packed, and column chromatography (PE) was performed to obtain 6g of colorless liquid, with a yield of 85.2%.
(2) Preparation of 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester
The above ethyl 4-nitro-1H-pyrrole-2-carboxylate (1 mmol,0.289g,1 eq) was dissolved in 20ml DMF, cesium carbonate (2 eq) was added, stirring at 50℃for 5min, the above 5- (3-bromopropyloxy) -2-chloro-1, 3-dimethylbenzene (2 eq) was added, after completion of the reaction, cooled to room temperature, 60ml of water was added, EA was extracted 3 times, the organic phase was combined, saturated brine wash, dried over sodium sulfate, the organic phase was suspended, and sand was produced, and column chromatography (EA: PE=20:1) gave 0.35g of colorless liquid, yield 76.4%.
(3) 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- (phenylamino) -1H-pyrrole-2-carboxylic acid ethyl ester
Ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4-nitro-1H-pyrrole-2-carboxylate (1.5 g,4.1 mmol) was dissolved in EtOH (50 ml) and then SnCl was added 2 ·2H 2 O (4.6 g,20 mmol). After the starting materials were reacted, they were cooled to room temperature and NaOH (2M) was added to ph=9. DCM (70 ml) was added and stirred for 15 min. The organic layer was separated, the aqueous layer extracted twice, the organic phases combined, na 2 SO 4 The organic phase was dried and suspended to give a dark brown oil. 1.0g of the crude product was dissolved in 30ml of toluene, and Pd was added to the solution 2 (dba) 3 (0.1 eq), BINAP (0.1 eq) and cesium carbonate (2 eq), 90 ℃ N 2 The heating was protected overnight. After completion of the reaction, cooling to room temperature, sand making, column chromatography (PE/ea=100:1 to80:1 v/v) purification gave a yellow liquid in 35% yield.
(4) Preparation of ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (phenyl) amino) -1H-pyrrole-2-carboxylate
NaH (2 eq) was added to a solution of ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4-nitro-1H-pyrrole-2-carboxylate in DMF (10 ml) under ice-bath conditions. After stirring for 10 minutes, 3-chlorobenzyl bromide (2 eq) was added and stirred at room temperature. After the reaction, 30ml of H was slowly added 2 O, ethyl acetate extraction 3 times, combining organic phases, organic phase with Na 2 SO 4 Drying and column chromatography (PE/ea=100:1 to 80:1 v/v) gave an oil in 54% yield.
(5) Preparation of the title compound
The above oil (0.3 g,0.58mmol,1 eq) was dissolved in ethanol/tetrahydrofuran mixture (7 ml/7 ml), naOH (2M, 10 eq) was added, heated and stirred overnight at 50℃and cooled to room temperature after completion of the reaction, 1M HCl was added to pH=1, and the mixture was taken overFiltering, washing the filter cake with water and drying to obtain light yellow solid 0.252g with a yield of 95%. mp is 208-209 ℃. 1 HNMR(300MHz,DMSO-d 6 )δ12.33(s,1H),7.37–7.19(m,4H),7.12–7.02(m,3H),6.82–6.62(m,6H),4.77(s,2H),4.43(t,J＝5.6Hz,2H),3.83(t,J＝5.1Hz,2H),2.27(s,6H),2.21–2.07(m,2H).HRMS(ESI):calcd for C 29 H 28 Cl 2 N 2 O 3 [M+H] + 523.1555,found 523.1546.HPLC(80:20methanol:water with 1‰TFA):t R ＝9.78min,96.6％.
Example 2:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (pyridin-2-yl) amino) -1H-pyrrole-2-carboxylic acid (Compound 2)
This was prepared from 2-bromopyridine, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, a brown solid, 76% yield, melting point: 139-140 ℃. 1 HNMR(300MHz,d6-DMSO)δ8.02(d,J＝5.3Hz,1H),7.73(s,1H),7.28(d,J＝5.2Hz,3H),7.18(s,2H),7.03–6.84(m,2H),6.80(s,1H),6.72(s,2H),5.04(s,2H),4.41(t,J＝6.1Hz,2H),3.82(t,J＝5.7Hz,2H),2.25(s,6H),2.10(q,J＝6.1Hz,2H).HRMS(ESI):calcd for C 28 H 27 Cl 2 N 3 O 3 [M+H] + 524.1502,found 524.1508.HPLC(80:20methanol:water with 1‰TFA):t R ＝5.95min,97.1％.
Example 3:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (naphthalen-1-yl) amino) -1H-pyrrole-2-carboxylic acid (compound 3)
The product was prepared from 1-bromonaphthalene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, whiteSolid, 65% yield, melting point: 118-120 ℃. 1 H NMR(300MHz,d6-DMSO)δ7.99(d,J＝8.1Hz,1H),7.91–7.87(m,1H),7.69(d,J＝8.0Hz,1H),7.49–7.37(m,4H),7.32(d,J＝7.4Hz,2H),7.29–7.17(m,2H),6.68(s,2H),6.34(d,J＝2.1Hz,1H),6.08(d,J＝2.0Hz,1H),4.73(s,2H),4.25(t,J＝6.5Hz,2H),3.72(t,J＝5.9Hz,2H),2.28(s,6H),2.00(p,J＝6.6Hz,2H).HRMS(ESI):calcd for C 33 H 30 Cl 2 N 2 O 3 [M+H] + 573.1712,found 573.1712.HPLC(80:20methanol:water with 1‰TFA):t R ＝13.9min,96.9％.
Example 4:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (o-tolyl) amino) -1H-pyrrole-2-carboxylic acid (Compound 4)
This was prepared from 2-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a brown solid with a yield of 72%, melting point: 109-110 DEG C 1 H NMR(300MHz,Chloroform-d)δ7.23–7.05(m,8H),6.80(d,J＝2.1Hz,1H),6.73–6.62(m,3H),4.75(s,2H),4.45(t,J＝6.4Hz,2H),3.91–3.81(m,2H),2.32–2.27(m,9H),2.20–2.11(m,2H).HRMS(ESI):calcd for C 30 H 30 Cl 2 N 2 O 3 [M+H] + 537.1712,found 537.1720.HPLC(80:20methanol:water with 1‰TFA):t R ＝10.2min,97.2％.
Example 5:
5- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -3- ((3-chlorobenzyl) (m-tolyl) amino) cyclopenta-1, 3-diene-1-carboxylic acid (Compound 5)
The product is prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane and 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester according to the following stepsPrepared as in example 1, white solid, 76% yield, melting point: 111-112 ℃. 1 HNMR(300MHz,d6-DMSO)δ12.30(s,1H),7.24(dq,J＝15.2,7.3,6.8Hz,4H),6.95(dd,J＝15.5,7.8Hz,2H),6.72(s,2H),6.68(s,1H),6.63–6.44(m,3H),4.72(s,2H),4.40(t,J＝5.6Hz,2H),3.80(t,J＝5.5Hz,2H),2.24(s,6H),2.12(s,5H). 13 C NMR(75MHz,d6-DMSO)δ162.06,154.41,148.60,142.44,138.47,134.51,133.57,130.79,130.68,129.23,127.88,127.04,126.91,126.77,126.63,125.71,125.57,125.40,124.27,122.15,120.89,120.41,119.29,113.64,112.16,105.29,65.23,46.11,31.00,21.85.HRMS(ESI):calcd for C 30 H 30 Cl 2 N 2 O 3 [M+H] + 537.1668,found 537.1682.HPLC(80:20methanol:water with 1‰TFA):t R ＝8.32min,97.6％.
Example 6:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (p-tolyl) amino) -1H-pyrrole-2-carboxylic acid (Compound 6)
This was prepared from 4-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a pale yellow solid with 74% yield, melting point: 117-118 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.31(s,1H),7.37–7.17(m,4H),6.96(s,1H),6.86(d,J＝8.3Hz,2H),6.78–6.65(m,5H),4.74(s,2H),4.41(t,J＝6.1Hz,2H),3.81(t,J＝5.3Hz,2H),2.28(s,6H),2.15(d,J＝7.8Hz,5H).HRMS(ESI):calcd for C 30 H 30 Cl 2 N 2 O 3 [M+H] + 537.1704,found 537.1712.HPLC(80:20methanol:water with 1‰TFA):t R ＝12.4min,98.1％.
Example 7:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (4-ethylphenyl) amino) -1H-pyrrole-2-carboxylic acid (Compound 7)
This was prepared from 4-ethyl bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, yellow solid, 82% yield, melting point: 121-122 ℃. 1 HNMR(300MHz,d6-DMSO)δ12.23(s,1H),7.23(dt,J＝21.8,7.5Hz,4H),6.94(d,J＝1.9Hz,1H),6.86(d,J＝8.4Hz,2H),6.75–6.60(m,5H),4.72(s,2H),4.38(t,J＝6.3Hz,2H),3.79(t,J＝5.9Hz,2H),2.44(q,J＝7.7Hz,2H),2.26(s,6H),2.11(p,J＝5.7,5.0Hz,2H),1.09(t,J＝7.6Hz,3H).HRMS(ESI):calcd for C 31 H 32 Cl 2 N 2 O 3 [M+H] + 551.1862,found 551.1868.HPLC(80:20methanol:water with 1‰TFA):t R ＝15.1min,96.5％.
Example 8:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (4-isopropylphenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 8)
This was prepared from 4-isopropyl bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a yellow solid with 64% yield, melting point: 124-125 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.28(s,1H),7.35–7.20(m,4H),6.98(d,J＝2.2Hz,1H),6.89(d,J＝8.7Hz,2H),6.81–6.68(m,5H),4.75(s,2H),4.41(t,J＝6.2Hz,2H),3.81(t,J＝5.7Hz,2H),2.73(p,J＝6.7Hz,1H),2.28(s,6H),2.15(q,J＝6.1Hz,2H),1.13(d,J＝6.9Hz,6H).HRMS(ESI):calcd for C 32 H 34 Cl 2 N 2 O 3 [M+H] + 565.2011,found 565.2025.HPLC(80:20methanol:water with 1‰TFA):t R ＝17.5min,98.3％.
Example 9:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (4-cyclopropylphenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 9)
This was prepared from 4-cyclopropyl-bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a yellow solid with 75% yield, melting point: 152-153 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.31(s,1H),7.39–7.17(m,4H),7.15–7.03(m,2H),6.82–6.64(m,7H),4.77(s,2H),4.43(t,J＝6.7Hz,2H),3.83(t,J＝6.0Hz,4H),2.28(d,J＝4.3Hz,8H),2.14(q,J＝6.4Hz,2H).HRMS(ESI):calcd for C 32 H 32 Cl 2 N 2 O 3 [M+H] + 563.1862,found 563.1868.HPLC(80:20methanol:water with 1‰TFA):t R ＝16.6min,96.3％.
Example 10:
4- ((4- (tert-butyl) phenyl) (3-chlorobenzyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (compound 10)
This was prepared from 4-tert-butylbromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a yellow solid with 67% yield, melting point: 179-180 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.23(s,1H),7.23(d,J＝17.6Hz,4H),7.12–6.91(m,3H),6.84–6.55(m,5H),4.73(s,2H),4.38(s,2H),3.79(s,2H),2.26(s,6H),2.11(s,2H),1.18(s,9H).HRMS(ESI):calcd for C 33 H 36 Cl 2 N 2 O 3 [M+H] + 579.2176,found 579.2181.HPLC(80:20methanol:water with 1‰TFA):t R ＝21.6min,95.9％.
Example 11:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-ethylphenyl) amino) -1H-pyrrole-2-carboxylic acid (Compound 11)
This was prepared from 3-ethyl bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, yellow solid, 76% yield, melting point: 106-107 ℃. 1 HNMR(300MHz,d6-DMSO)δ12.24(s,1H),7.26(dd,J＝26.7,6.2Hz,4H),7.02–6.91(m,2H),6.72(s,2H),6.68(d,J＝2.0Hz,1H),6.63(s,1H),6.55(dd,J＝13.2,7.9Hz,2H),4.73(s,2H),4.40(t,J＝6.4Hz,2H),3.81(t,J＝5.9Hz,2H),2.47–2.34(m,2H),2.24(s,6H),2.11(p,J＝6.1Hz,2H),1.05(t,J＝7.6Hz,3H).HRMS(ESI):calcd for C 31 H 32 Cl 2 N 2 O 3 [M+H] + 551.1866,found 551.1868.HPLC(80:20methanol:water with 1‰TFA):t R ＝14.7min,96.7％.
Example 12:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-isopropylphenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 12)
This was prepared from 4-isopropyl bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a yellow solid with 77% yield, melting point: 121-122 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.25(s,1H),7.47–7.15(m,4H),7.11–6.93(m,2H),6.71(q,J＝10.3,7.1Hz,3H),6.57(d,J＝7.1Hz,2H),4.74(s,2H),4.40(s,2H),3.82(s,2H),2.77–2.59(m,1H),2.25(s,5H),2.12(s,2H),1.07(d,J＝6.6Hz,6H).HRMS(ESI):calcd for C 32 H 32 Cl 2 N 2 O 3 [M+H] + 565.2018,found565.2025.HPLC(80:20methanol:water with 1‰TFA):t R ＝14.8min,97.2％.
Example 13:
4- ((3- (tert-butyl) phenyl) (3-chlorobenzyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (compound 13)
This was prepared from 3-tert-butylbromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a yellow solid with a yield of 68%, melting point: 107-109 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.31(s,2H),7.24–7.18(m,2H),7.10(t,J＝7.9Hz,1H),7.01(s,1H),6.95(s,1H),6.88(d,J＝7.5Hz,1H),6.77(s,1H),6.71(d,J＝7.8Hz,1H),6.63(s,2H),4.73(s,2H),4.49(d,J＝5.7Hz,2H),3.85(t,J＝4.5Hz,2H),2.35(s,6H),2.27(s,2H),1.27(s,9H).HRMS(ESI):calcd for C 33 H 36 Cl 2 N 2 O 3 [M+H] + 579.2174,found 579.2181.HPLC(80:20methanol:water with 1‰TFA):t R ＝17.4min,98.1％.
Example 14:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-chlorophenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 14)
This was prepared from 3-chlorobromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a white solid with 71% yield, melting point: 132-133 ℃. 1 HNMR(300MHz,d6-DMSO)δ12.38(s,1H),7.38–7.14(m,4H),7.05(s,3H),6.80–6.62(m,5H),4.73(s,2H),4.43(s,2H),3.78(s,2H),2.27(s,6H),2.19–2.06(m,2H).HRMS(ESI):calcd for C 29 H 27 Cl 3 N 2 O 3 [M+H] + 559.1127,found 559.1136.HPLC(80:20methanol:water with 1‰TFA):t R ＝11.5min,97.6％.
Example 15:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-fluorophenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 15)
This was prepared from 3-fluorobromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a yellow solid with a yield of 68%, melting point: 122-123 ℃. 1 HNMR(300MHz,d6-DMSO)δ7.28(d,J＝10.4Hz,3H),7.18(d,J＝7.0Hz,1H),7.11–7.01(m,2H),6.73(d,J＝2.1Hz,1H),6.71(s,2H),6.56–6.38(m,3H),4.75(s,2H),4.42(t,J＝6.4Hz,2H),3.81(t,J＝5.9Hz,2H),2.23(s,6H),2.12(p,J＝5.9Hz,2H).HRMS(ESI):calcd for C 29 H 27 Cl 2 FN 2 O 3 [M+H] + 541.1469,found541.1461.HPLC(80:20methanol:water with 1‰TFA):t R ＝9.93min,95.7％.
Example 16:
4- ((3-Acetylphenyl) (3-chlorobenzyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (Compound 16)
This was prepared from 3-bromoacetophenone, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, yellow solid, 43% yield, melting point: 125-126 ℃. 1 H NMR(300MHz,Chloroform-d)δ7.41(s,1H),7.29(d,J＝8.6Hz,2H),7.18(t,J＝12.8Hz,6H),6.96(s,1H),6.70(s,1H),6.56(s,2H),4.70(s,2H),4.46(s,2H),3.76(s,2H),2.49(s,3H),2.29(s,6H),2.20(s,1H).HRMS(ESI):calcd for C 31 H 30 Cl 2 N 2 O 4 [M+H] + 565.1661,found 565.1652.HPLC(80:20methanol:water with1‰TFA):t R ＝7.05min,96.3％.
Example 17:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-nitrophenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 17)
This was prepared from 3-nitrobromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a red solid with 75% yield, melting point: 162-163 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.59(dd,J＝4.2,1.8Hz,2H),7.25(t,J＝3.7Hz,4H),7.13(dd,J＝6.5,3.0Hz,1H),7.07(d,J＝1.9Hz,1H),7.04–6.98(m,1H),6.86(d,J＝1.9Hz,1H),6.62(s,2H),4.76(s,2H),4.57(t,J＝6.3Hz,2H),3.85(t,J＝5.4Hz,2H),2.32(s,8H).HRMS(ESI):calcd for C 29 H 27 Cl 2 N 3 O 5 [M+H] + 568.1396,found 568.1406.HPLC(80:20methanol:water with 1‰TFA):t R ＝8.4min,95.1％.
Example 18:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-cyanophenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 18)
This was prepared from 3-nitrile bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, yellow solid, 67% yield, melting point: 136-137 deg.c. 1 H NMR(300MHz,CDCl 3 )δ7.28–7.14(m,4H),7.09–6.92(m,5H),6.83(s,1H),6.62(s,2H),4.70(s,2H),4.56(s,2H),3.84(s,2H),2.34(s,6H),2.30(s,2H).HRMS(ESI):calcd for C 30 H 27 Cl 2 N 3 O 3 [M+H] + 548.1495,found548.1508.HPLC(80:20methanol:water with 1‰TFA):t R ＝6.8min,95.4％.
Example 19:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-methoxyphenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 19)
This was prepared from 3-methoxybromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, as a yellow solid with 78% yield, melting point: 121-122 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.37(s,1H),7.27(h,J＝7.4,6.8Hz,4H),7.07(s,1H),6.98(t,J＝8.3Hz,1H),6.74(s,3H),6.36(d,J＝8.7Hz,1H),6.28(s,2H),4.74(s,2H),4.50–4.36(m,2H),3.82(t,J＝5.0Hz,2H),3.62(s,3H),2.26(s,6H),2.13(t,J＝6.6Hz,2H). 13 C NMR(75MHz,d6-DMSO)δ161.97,160.57,156.88,149.76,142.36,136.94,133.61,130.67,130.55,130.09,127.06,126.72,125.56,125.52,124.80,120.96,115.04,114.03,107.55,103.10,100.97,55.80,55.12,45.79,30.61,20.87.HRMS(ESI):calcd for C 30 H 30 Cl 2 N 2 O 4 [M+H] + 553.1656,found 553.1661.HPLC(80:20methanol:water with1‰TFA):t R ＝10.4min,97.8％.
Example 20:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3-hydroxyphenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 20)
(1) Preparation of 4- ((3- (benzyloxy) phenyl) (3-chlorobenzyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid
This product was prepared from 3-benzyloxy bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester as in example 1, brown oil with a yield of 64%, 1 HNMR(300MHz,Chloroform-d)δ7.42(ddq,J＝7.0,1.8,1.0Hz,2H),7.38–7.29(m,8H),7.27–7.20(m,2H),6.99(dt,J＝7.5,1.5Hz,1H),6.72(dt,J＝7.5,1.5Hz,1H),6.69(s,2H),6.63(t,J＝1.5Hz,1H),5.05(t,J＝0.9Hz,2H),5.01(t,J＝1.0Hz,2H),4.03(dt,J＝16.9,7.0Hz,4H),2.29(s,6H),2.08(p,J＝7.1Hz,2H).ESI-MS m/z:627.2[M+H] + .
(2) Preparation of the title compound
The above 4- ((3- (benzyloxy) phenyl) (3-chlorobenzyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid was dissolved in ethanol, palladium on carbon (0.05 eq) was added, hydrogen was replaced three times, heated at 60 ℃ for 3 hours, cooled to room temperature and filtered, the filtrate was suspended to give the title product as a grey solid, yield 64%, melting point: 121-122 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.27–7.11(m,4H),7.05–6.94(m,2H),6.78(s,1H),6.62(s,2H),6.42–6.24(m,3H),4.69(s,2H),4.49(t,J＝5.6Hz,2H),3.81(t,J＝5.0Hz,2H),2.34(s,6H),2.29–2.19(m,2H).HRMS(ESI):calcd for C 29 H 28 Cl 2 N 2 O 4 [M+H] + 539.1504,found 539.1500.HPLC(80:20methanol:water with 1‰TFA):t R ＝7.26min,96.4％.
Example 21:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (3- (1-hydroxyethyl) phenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 21)
Example 16 (0.2 g) above was dissolved in THF/DCM (1:1, 10 ml) and NaBH4 (2 eq) was added. After the reaction, HCl (2M) was added under ice bath conditions, the mixture was extracted 3 times with DCM, the organic phases were combined, the organic phase was dried over saturated brine and sodium sulfate, and suspended to give a brown solid with a yield of 56%, melting point: 131-132 ℃. 1 H NMR(300MHz,DMSO-d 6 )δ12.30(s,1H),7.33–7.19(m,4H),7.06–6.98(m,2H),6.85(s,1H),6.75(s,2H),6.72–6.60(m,3H),4.76(s,2H),4.62–4.51(m,1H),4.42(t,J＝6.4Hz,2H),3.83(t,J＝5.9Hz,2H),2.27(s,6H),2.19–2.07(m,2H),1.23(d,J＝6.4Hz,3H).HRMS(ESI):calcd for C 31 H 32 Cl 2 N 2 O 4 [M+H] + 567.1817,found 567.1820.HPLC(80:20methanol:water with 1‰TFA):t R ＝6.04min,97.1％.
Example 22:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((thiophen-3-ylmethyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 22)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-bromomethylthiophene according to example 1 as a yellow solid with a yield of 56%, melting point: 137-138 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.20(s,1H),7.41(dd,J＝4.8,3.0Hz,1H),7.18(s,1H),7.02–6.90(m,3H),6.74(d,J＝3.7Hz,2H),6.72–6.54(m,3H),6.45(dd,J＝13.3,7.1Hz,1H),4.68(s,2H),4.40(t,J＝6.2Hz,2H),3.83(t,J＝5.9Hz,2H),2.26(s,6H),2.13(d,J＝4.1Hz,5H).HRMS(ESI):calcd for C 28 H 29 ClN 2 O 3 [M+H] + 509.1657,found 509.1666.HPLC(80:20methanol:water with 1‰TFA):t R ＝9.22min,97.6％.
Example 23:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((naphthalen-1-ylmethyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 23)
The product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 1-bromomethylnaphthalene according to example 1, 67% yield, melting point: 91.0-92.4 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.20(s,1H),8.10–7.69(m,3H),7.44(dd,J＝58.0,4.8Hz,4H),7.08–6.84(m,2H),6.82–6.19(m,6H),5.22(s,2H),4.37(s,2H),3.80(s,2H),2.17(d,J＝37.3Hz,11H).HRMS(ESI):calcd for C 34 H 33 ClN 2 O 3 [M+H] + 553.2246,found 553.2258.HPLC(80:20methanol:water with 1‰TFA):t R ＝15.2min,96.5％.
Example 24:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((naphthalen-2-ylmethyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 24)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 2-bromomethylnaphthalene as in example 1, as a white solid with a yield of 72%, melting point: 101-102 ℃. 1 H NMR(300MHz,DMSO-d 6 )δ12.20(s,1H),7.92–7.69(m,4H),7.52–7.36(m,3H),7.05(s,1H),6.94(t,J＝7.8Hz,1H),6.85–6.52(m,5H),6.48(d,J＝6.8Hz,1H),4.89(s,2H),4.40(t,J＝6.2Hz,2H),3.81(t,J＝5.9Hz,2H),2.23(s,6H),2.12(s,5H).HRMS(ESI):calcd for C 34 H 33 ClN 2 O 3 [M+H] + 553.2246,found 553.2248.HPLC(80:20methanol:water with 1‰TFA):t R ＝15.4min,97.2％.
Example 25:
4- (benzyl (m-tolyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (compound 25)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester and benzyl bromide as in example 1, as a white solid with 67% yield, melting point: 149-150 ℃. 1 HNMR(300MHz,d6-DMSO)δ12.23(s,1H),7.25(d,J＝6.2Hz,5H),6.94(dd,J＝14.9,7.3Hz,2H),6.70(d,J＝13.2Hz,3H),6.67–6.54(m,2H),6.47(d,J＝7.2Hz,1H),4.73(s,2H),4.39(t,J＝5.8Hz,2H),3.81(t,J＝5.6Hz,2H),2.25(s,6H),2.12(s,5H).HRMS(ESI):calcd for C 30 H 31 ClN 2 O 3 [M+H] + 503.2095,found 503.2101.HPLC(80:20methanol:water with 1‰TFA):t R ＝10.9min,96.7％.
Example 26:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((2-chlorobenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 26)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-chlorobenzyl bromide according to example 1 as a yellow solid with a yield of 65%, melting point: 134-135 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.22(s,1H),7.33(d,J＝7.4Hz,1H),7.21–7.07(m,3H),6.95(d,J＝2.1Hz,1H),6.85(t,J＝7.8Hz,1H),6.62(d,J＝2.9Hz,3H),6.45–6.33(m,3H),4.65(s,2H),4.29(t,J＝6.7Hz,2H),3.70(t,J＝6.0Hz,2H),2.14(s,6H),2.02(s,5H).HRMS(ESI):calcd for C 30 H 30 Cl 2 N 2 O 3 [M+H] + 537.1705,found 537.1702.HPLC(80:20methanol:water with 1‰TFA):t R ＝14.1min,98.1％.
Example 27:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((2-methylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 27)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 2-methyl bromobenzyl according to example 1 as a white solid with a yield of 56%, melting point: 121-122 ℃. 1 H NMR(300MHz,Chloroform-d)δ7.28(d,J＝7.5Hz,1H),7.20–7.16(m,2H),7.12–7.00(m,3H),6.78(d,J＝1.8Hz,1H),6.71–6.59(m,5H),4.72(s,2H),4.48(t,J＝6.3Hz,2H),3.84(t,J＝5.4Hz,2H),2.36(s,6H),2.30(s,3H),2.27(s,3H),2.24(d,J＝5.8Hz,2H).HRMS(ESI):calcd for C 31 H 33 ClN 2 O 3 [M+H] + 537.1705,found 537.1702.HPLC(80:20methanol:water with 1‰TFA):t R ＝13.0min,97.6％.
Example 28:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-methylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 28)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-methyl bromobenzyl as in example 1 as a yellow solid with a yield of 72%, melting point: 139-140 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.32(s,1H),7.18(t,J＝7.5Hz,1H),7.12–6.92(m,5H),6.77(s,2H),6.72(d,J＝2.1Hz,1H),6.66–6.55(m,2H),6.51(d,J＝7.4Hz,1H),4.72(s,2H),4.43(t,J＝6.4Hz,2H),3.85(t,J＝6.0Hz,2H),2.28(s,9H),2.16(s,5H).HRMS(ESI):calcd for C 31 H 33 ClN 2 O 3 [M+H] + 517.2248,found 517.2258.HPLC(80:20methanol:water with 1‰TFA):t R ＝9.84min,98.5％.
Example 29:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((4-chlorobenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 29)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4-chlorobenzyl bromide according to example 1 as a yellow solid, 57% yield, melting point: 141-142 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.33(s,1H),7.33(d,J＝7.4Hz,2H),7.26(d,J＝8.2Hz,2H),7.04–6.93(m,2H),6.76(s,2H),6.72(s,1H),6.65–6.48(m,3H),4.74(s,2H),4.48–4.38(m,2H),3.88–3.78(m,2H),2.28(s,6H),2.15(s,5H).HRMS(ESI):calcd for C 30 H 30 Cl 2 N 2 O 3 [M+H] + 537.1706,found537.1702.HPLC(80:20methanol:water with 1‰TFA):t R ＝12.4min,97.6％.
Example 30:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((4-methylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 30)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4-methyl bromobenzyl as in example 1, as a white solid with a yield of 66%, melting point: 128-129 ℃. 1 H NMR(300MHz,d6-DMSO)δ7.10(d,J＝7.9Hz,2H),7.04(d,J＝7.9Hz,2H),6.98–6.89(m,2H),6.73(s,2H),6.67(d,J＝2.1Hz,1H),6.62–6.53(m,2H),6.46(d,J＝7.4Hz,1H),4.67(s,2H),4.39(t,J＝6.5Hz,2H),3.81(t,J＝6.1Hz,2H),2.25(s,6H),2.23(s,3H),2.10(d,J＝8.0Hz,5H).HRMS(ESI):calcd for C 31 H 33 ClN 2 O 3 [M+H] + 517.2253,found 517.2258.HPLC(80:20methanol:water with1‰TFA):t R ＝12.8min,98.1％.
Example 31:
4- ((3-bromobenzyl) (m-tolyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (compound 31)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-bromobenzyl according to example 1 as a yellow solid, 63% yield, melting point: 141-142 ℃. 1 H NMR(300MHz,d6-DMSO)δ7.42(d,J＝10.8Hz,2H),7.25(s,2H),6.99(d,J＝14.0Hz,2H),6.72(d,J＝15.3Hz,3H),6.67–6.48(m,3H),4.75(s,2H),4.43(s,2H),3.83(s,2H),2.27(s,6H),2.15(s,5H).HRMS(ESI):calcd for C 30 H 30 BrClN 2 O 3 [M+H] + 583.1180,found 583.1186.HPLC(80:20methanol:water with 1‰TFA):t R ＝13.5min,97.8％.
Example 32:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-methoxybenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 32)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-methoxybromobenzyl according to example 1 as a yellow solid, yield 79%, melting point: 145-146 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.22(s,1H),7.10(d,J＝7.9Hz,2H),7.04(d,J＝8.0Hz,2H),6.99–6.94(m,1H),6.91(d,J＝7.8Hz,1H),6.73(s,2H),6.67(d,J＝2.1Hz,1H),6.63–6.53(m,2H),6.46(d,J＝7.4Hz,1H),4.67(s,2H),4.39(t,J＝6.5Hz,2H),3.81(t,J＝6.0Hz,2H),2.25(s,6H),2.23(s,3H),2.11(s,5H).HRMS(ESI):calcd for C 31 H 33 ClN 2 O 4 [M+H] + 533.2207,found 533.2203.HPLC(80:20methanol:water with 1‰TFA):t R ＝12.5min,96.2％.
Example 33:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-fluorobenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 33)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-fluorobenzyl bromide according to example 1 as a yellow solid, yield 83%, melting point: 146-147 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.23(s,1H),7.37–7.25(m,1H),7.13–6.89(m,5H),6.72(s,2H),6.68(d,J＝2.1Hz,1H),6.65–6.42(m,3H),4.74(s,2H),4.40(t,J＝6.3Hz,2H),3.81(t,J＝6.0Hz,2H),2.25(s,6H),2.11(d,J＝9.6Hz,5H). 13 C NMR(75MHz,d6-DMSO)δ161.96,156.90,148.63,142.93,138.42,136.95,130.80,130.76,129.19,125.54,124.59,122.90,120.84,119.16,115.29,115.06,113.95,113.82,113.71,113.43,112.04,64.91,55.77,45.74,30.57,21.82,20.87.HRMS(ESI):calcd for C 30 H 30 ClFN 2 O 3 [M+H] + 521.1998,found 521.2007.HPLC(80:20methanol:water with1‰TFA):t R ＝9.39min,96.2％.
Example 34:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- (m-tolyl (3- (trifluoromethyl) benzyl) amino) -1H-pyrrole-2-carboxylic acid (compound 34)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-trifluoromethyl bromobenzyl according to example 1 as an orange solid with 67% yield, melting point: 121-122 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.20(s,1H),7.43(q,J＝9.4,7.8Hz,4H),6.95–6.80(m,3H),6.60(d,J＝11.0Hz,2H),6.54–6.34(m,3H),4.72(s,2H),4.29(s,2H),3.69(s,2H),2.13(s,6H),2.02(s,5H).HRMS(ESI):calcd for C 31 H 30 ClF 3 N 2 O 3 [M+H] + 571.1972,found 571.1975.HPLC(80:20methanol:water with 1‰TFA):t R ＝12.6min,97.4％.
Example 35:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- (m-tolyl (3- (trifluoromethoxy) benzyl) amino) -1H-pyrrole-2-carboxylic acid (compound 35)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-trifluoromethoxybromide according to example 1, as a red solid, 59% yield, melting point: 104-105 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.30(s,1H),7.40(t,J＝7.8Hz,1H),7.26(d,J＝7.7Hz,1H),7.17(d,J＝7.9Hz,2H),7.02–6.91(m,2H),6.72(s,2H),6.67(d,J＝2.1Hz,1H),6.63–6.45(m,3H),4.77(s,2H),4.39(t,J＝6.3Hz,2H),3.80(t,J＝6.1Hz,2H),2.24(s,6H),2.12(s,5H).HRMS(ESI):calcd for C 31 H 30 ClF 3 N 2 O 4 [M+H] + 587.1917,found 587.1924.HPLC(80:20methanol:water with 1‰TFA):t R ＝10.8min,96.5％.
Example 36:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-formylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 36)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-formylbromobenzyl according to example 1 as a white solid with a yield of 69%, melting point: 106-107 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.19(s,1H),9.95(s,1H),7.82–7.69(m,2H),7.52(dt,J＝15.0,7.4Hz,2H),6.96(dd,J＝16.7,9.0Hz,2H),6.70(s,3H),6.63–6.54(m,2H),6.49(d,J＝7.1Hz,1H),4.82(s,2H),4.39(t,J＝5.8Hz,2H),3.79(t,J＝5.3Hz,2H),2.23(s,6H),2.10(d,J＝11.7Hz,5H).HRMS(ESI):calcd for C 31 H 31 ClN 2 O 4 [M+H] + 531.2043,found 531.2051.HPLC(80:20methanol:water with 1‰TFA):t R ＝6.99min,97.8％.
Example 37:
- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((4-isopropylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 37)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4-isopropyl bromobenzyl as in example 1, as a white solid,yield 67%, melting point: 118-119 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.30(s,1H),7.14(s,4H),7.08–6.89(m,2H),6.76(s,2H),6.70(d,J＝2.0Hz,1H),6.63(s,1H),6.58(d,J＝8.3Hz,1H),6.48(d,J＝7.4Hz,1H),4.70(s,2H),4.41(t,J＝6.6Hz,2H),3.83(t,J＝6.0Hz,2H),2.81(dq,J＝13.9,6.9Hz,1H),2.26(s,6H),2.14(s,5H),1.18(s,3H),1.16(s,3H).HRMS(ESI):calcd for C 33 H 37 ClN 2 O 3 [M+H] + 545.2565,found 545.2571.HPLC(80:20methanol:water with 1‰TFA):t R ＝18.3min,98.2％.
Example 38:
4- ((4- (tert-butyl) benzyl) (m-tolyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (compound 38)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4-tert-butylbromobenzyl as in example 1 as a yellow solid with a yield of 67%, melting point: 123-124 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.31(s,1H),7.29(d,J＝7.9Hz,2H),7.16(d,J＝8.0Hz,2H),7.02(d,J＝2.1Hz,1H),6.95(t,J＝7.8Hz,1H),6.76(s,2H),6.71(d,J＝2.0Hz,1H),6.64(s,1H),6.58(d,J＝8.5Hz,1H),6.48(d,J＝7.4Hz,1H),4.71(s,2H),4.48–4.36(m,2H),3.83(t,J＝6.1Hz,2H),2.26(s,6H),2.14(s,5H),1.25(s,9H).HRMS(ESI):calcd for C 34 H 39 ClN 2 O 3 [M+H] + 559.2724,found559.2727.HPLC(80:20methanol:water with 1‰TFA):t R ＝21.1min,97.3％.
Example 39:
4- (([ [1,1' -biphenyl ] -4-ylmethyl) (m-tolyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (compound 39)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4-tert-butylbromobenzyl as in example 1, as a pale yellow solid with a yield of 65%, melting point: 104-105 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.23(s,1H),7.57(dd,J＝16.0,7.8Hz,4H),7.43(t,J＝7.5Hz,2H),7.33(t,J＝8.5Hz,3H),7.01(d,J＝2.1Hz,1H),6.95(t,J＝7.8Hz,1H),6.72(d,J＝2.7Hz,3H),6.67–6.56(m,2H),6.48(d,J＝7.5Hz,1H),4.78(s,2H),4.40(t,J＝6.5Hz,2H),3.81(t,J＝6.0Hz,2H),2.23(s,6H),2.13(s,5H).HRMS(ESI):calcd for C 36 H 35 ClN 2 O 3 [M+H] + 579.2404,found 579.2414.HPLC(80:20methanol:water with 1‰TFA):t R ＝16.8min,96.8％.
Example 40:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((4-ethynylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 40)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4-ethynyl bromobenzyl according to example 1 as a white solid with 64% yield, melting point: 142-143 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.35(s,1H),7.42(d,J＝7.8Hz,2H),7.29(d,J＝8.0Hz,2H),7.08–6.95(m,2H),6.79(s,2H),6.75(d,J＝2.1Hz,1H),6.69–6.51(m,3H),4.79(s,2H),4.46(s,2H),4.18(s,1H),3.86(s,2H),2.31(s,6H),2.18(s,5H). 13 C NMR(75MHz,d6-DMSO)δ161.96,156.91,148.63,140.86,138.41,136.97,132.21,130.76,129.19,127.24,125.54,124.63,120.79,120.38,119.05,115.20,115.07,113.90,111.98,83.92,80.85,64.88,55.94,45.73,30.55,21.85,20.90.HRMS(ESI):calcd for C 32 H 31 ClN 2 O 3 [M+H] + 527.2094,found 527.2101.HPLC(80:20methanol:water with1‰TFA):t R ＝9.34min,97.1％.
Example 41:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((4- (prop-1-yn-1-yl) benzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 41)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4-propynylbromobenzyl as in example 1 as a white solid with a yield of 64%, melting point: 136-137 deg.c. 1 H NMR(300MHz,d6-DMSO)δ12.32(s,1H),7.29(d,J＝8.1Hz,2H),7.20(d,J＝7.9Hz,2H),7.04–6.91(m,2H),6.75(s,2H),6.71(s,1H),6.64–6.53(m,2H),6.49(d,J＝7.3Hz,1H),4.73(s,2H),4.42(t,J＝6.4Hz,2H),3.82(t,J＝6.0Hz,2H),2.27(s,6H),2.14(s,5H),2.04(s,3H).HRMS(ESI):calcd for C 33 H 33 ClN 2 O 3 [M+H] + 541.2258,found 541.2258.HPLC(80:20methanol:water with 1‰TFA):t R ＝12.7min,98.5％.
Example 42:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3, 4-dimethylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 42)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3, 4-dimethylbenzyl bromide according to example 1 as a white solid, 75% yield, melting point: 136-137 deg.c. 1 H NMR(300MHz,d6-DMSO)δ12.14(s,1H),7.02–6.89(m,5H),6.72(s,2H),6.67(d,J＝2.1Hz,1H),6.60(s,1H),6.55(dd,J＝7.9,2.4Hz,1H),6.46(d,J＝7.4Hz,1H),4.64(s,2H),4.39(t,J＝6.5Hz,2H),3.81(t,J＝6.0Hz,2H),2.14(s,6H),2.11(s,5H).HRMS(ESI):calcd for C 32 H 35 ClN 2 O 3 [M+H] + 531.2404,found 531.2414.HPLC(80:20methanol:water with 1‰TFA):t R ＝9.78min,97.3％.
Example 43:
4- ((3-bromo-4-methylbenzyl) (m-tolyl) amino) -1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -1H-pyrrole-2-carboxylic acid (compound 43)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-bromo-4-methylbenzyl bromide according to example 1 as a yellow solid, 77% yield, melting point: 147-148 ℃. 1 HNMR(300MHz,d6-DMSO)δ12.28(s,1H),7.42(s,1H),7.20(d,J＝7.8Hz,1H),7.12(d,J＝7.8Hz,1H),6.98(d,J＝2.0Hz,1H),6.94(t,J＝7.8Hz,1H),6.71(s,2H),6.68–6.38(m,4H),4.69(s,2H),4.40(t,J＝6.5Hz,2H),3.81(t,J＝6.0Hz,2H),2.26(d,J＝8.3Hz,9H),2.12(s,5H).HRMS(ESI):calcd for C 31 H 32 BrClN 2 O 3 [M+H] + 597.1335,found 597.1343.HPLC(80:20methanol:water with 1‰TFA):t R ＝16.2min,98.1％.
Example 44:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3, 4-dichlorobenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 44)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3, 4-dichlorobenzyl bromide as in example 1, as a yellow solid, 57% yield, melting point: 143-144 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.24(s,1H),7.55–7.40(m,2H),7.20(d,J＝8.3Hz,1H),7.05–6.90(m,2H),6.70(d,J＝6.7Hz,3H),6.62–6.43(m,3H),4.72(s,2H),4.40(t,J＝6.2Hz,2H),3.80(t,J＝5.8Hz,2H),2.24(s,6H),2.13(s,5H).HRMS(ESI):calcd for C 30 H 29 Cl 3 N 2 O 3 [M+H] + 573.1293,found573.1293.HPLC(80:20methanol:water with 1‰TFA):t R ＝15.1min,97.7％.
Example 45:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3, 5-dimethylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (Compound 45)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3, 5-dimethylbenzyl bromide according to example 1 as a yellow solid, 53% yield, melting point: 161-162 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.08(tt,J＝14.0,7.4Hz,5H),6.81(s,1H),6.67(d,J＝16.7Hz,4H),4.76(s,2H),4.48(t,J＝6.4Hz,2H),3.86(t,J＝5.8Hz,2H),2.36(s,9H),2.27(s,3H),2.20(s,5H).HRMS(ESI):calcd for C 32 H 35 ClN 2 O 3 [M+H] + 531.2409,found 531.2414.HPLC(80:20methanol:water with1‰TFA):t R ＝15.0min,98.5％.
Example 46:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((2, 3-dimethylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 46)
This product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 2, 3-dimethylbenzyl bromide according to example 1, 75% yield, melting point: 161-162 ℃. 1 H NMR(300MHz,Chloroform-d)δ7.14(d,J＝7.2Hz,1H),7.10–6.95(m,4H),6.79(d,J＝1.9Hz,1H),6.68(d,J＝4.7Hz,1H),6.62(d,J＝4.7Hz,3H),4.75(s,2H),4.53–4.42(m,2H),3.85(t,J＝5.6Hz,2H),2.36(s,9H),2.26(s,3H),2.23(d,J＝6.1Hz,2H),2.20(s,3H).HRMS(ESI):calcd for C 32 H 35 ClN 2 O 3 [M+H] + 531.2414,found 531.2399.HPLC(80:20methanol:water with 1‰TFA):t R ＝14.5min,97.4％.
Example 47:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((2, 3-difluorobenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 47)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 2, 3-difluorobromobenzyl as in example 1 as a yellow solid, yield: 96%, melting point: 148.0-151.3 ℃. 1 HNMR(300MHz,CDCl 3 )δ7.14–6.94(m,5H),6.76(d,J＝2.0Hz,1H),6.67(d,J＝7.5Hz,3H),6.63(s,2H),4.68(s,2H),4.51(t,J＝6.4Hz,2H),3.88–3.79(m,2H),2.35(s,6H),2.27(s,5H).HRMS(ESI):calcd for C 30 H 29 ClF 2 N 2 O 3 [M+H] + 539.1903,found 539.1913.HPLC(80:20methanol:water with1‰TFA):t R ＝9.34min,98.2％.
Example 48:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((2, 4-dichlorobenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 48)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 2, 4-dichlorobenzyl bromide as in example 1 as a yellow solid, 58% yield, melting point: 137-138 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.40(d,J＝2.0Hz,1H),7.27(d,J＝8.3Hz,1H),7.15–7.00(m,3H),6.81(d,J＝2.3Hz,1H),6.63(d,J＝6.3Hz,5H),4.79(s,2H),4.51(t,J＝6.5Hz,2H),3.84(t,J＝5.6Hz,2H),2.35(s,6H),2.27(s,5H).HRMS(ESI):calcd for C 30 H 29 Cl 3 N 2 O 3 [M+H] + 573.1289,found 573.1293.HPLC(80:20methanol:water with 1‰TFA):t R ＝23.67min,97.0％.
Example 49:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((2, 6-dichlorobenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 49)
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 2, 6-dichlorobenzyl bromide as in example 1 as a yellow solid, 71% yield, melting point: 148-149 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.35(d,J＝2.4Hz,1H),7.32(s,1H),7.29(s,1H),7.17(dd,J＝8.5,2.5Hz,1H),7.10–7.00(m,2H),6.82(d,J＝2.1Hz,1H),6.64(d,J＝12.2Hz,5H),4.79(s,2H),4.52(t,J＝6.5Hz,2H),3.85(t,J＝5.7Hz,2H),2.35(s,6H),2.29(d,J＝4.5Hz,5H).HRMS(ESI):calcd for C 30 H 29 Cl 3 N 2 O 3 [M+H] + 573.1325,found 573.1322.HPLC(80:20methanol:water with 1‰TFA):t R ＝16.8min,98.7％.
Example 50:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((((1-phenyl-1H-1, 2, 3-triazol-4-yl) methyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 50)
(1) Preparation of ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- (prop-2-yn-1-yl (m-tolyl) amino) -1H-pyrrole-2-carboxylate
The product was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 3-bromoprop-1-yne according to the above preparation method of ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chlorobenzyl) (phenyl) amino) -1H-pyrrole-2-carboxylate, as a yellow oil, with a yield of 56%. 1 H NMR(300MHz,Chloroform-d)δ7.28–7.22(m,2H),7.13(dt,J＝7.5,1.6Hz,1H),6.93(t,J＝1.5Hz,1H),6.88(ddq,J＝7.3,1.5,0.7Hz,1H),6.69(s,2H),6.61(d,J＝1.5Hz,1H),4.78(d,J＝3.1Hz,2H),4.29(q,J＝8.0Hz,2H),4.04(dt,J＝8.8,7.1Hz,4H),2.71(t,J＝2.9Hz,1H),2.37(d,J＝0.7Hz,3H),2.33(s,6H),2.10(p,J＝7.1Hz,2H),1.35(t,J＝8.0Hz,3H).ESI-MS m/z:479.2[M+H] + .
(2) Preparation of ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((((1-phenyl-1H-1, 2, 3-triazol-4-yl) methyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylate
Ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- (prop-2-yn-1-yl (m-tolyl) amino) -1H-pyrrole-2-carboxylate (0.3 g), DIPEA (2 eq), cuI (0.1 eq) and azidobenzene (2 eq) were dissolved in methanol. Under nitrogen protection, heating at 50deg.C for 2 hours, cooling to room temperature after the reaction, sand making, column chromatography separation (PE/EA=20:1 to 10:1 v/v) to obtain brown oil with 73% yield. 1 H NMR(300MHz,Chloroform-d)δ8.58(s,1H),7.75–7.65(m,2H),7.52–7.46(m,2H),7.31–7.13(m,4H),6.93(q,J＝1.1Hz,1H),6.85(dtd,J＝7.3,1.5,0.8Hz,1H),6.69(s,2H),6.58(d,J＝1.5Hz,1H),5.24(s,2H),4.28(q,J＝8.0Hz,2H),4.04(td,J＝7.0,5.5Hz,4H),2.37(t,J＝0.6Hz,3H),2.29(s,6H),2.08(p,J＝7.1Hz,2H),1.35(t,J＝8.0Hz,3H).ESI-MS m/z:598.2[M+H] + .
(3) Preparation of the title compound
This was prepared from the above ethyl 1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- (prop-2-yn-1-yl (m-tolyl) amino) -1H-pyrrole-2-carboxylate as in example 1, as a white solid, 76% yield, melting point: 177-178 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.32(s,1H),8.63(s,1H),7.94–7.80(m,2H),7.58(t,J＝7.5Hz,2H),7.48(t,J＝7.3Hz,1H),7.13(s,1H),6.98(t,J＝7.4Hz,1H),6.82(s,1H),6.72(d,J＝9.9Hz,4H),6.55–6.46(m,1H),4.83(s,2H),4.53–4.35(m,2H),3.95–3.76(m,2H),2.23(s,6H),2.17(s,5H). 13 C NMR(75MHz,d6-DMSO)δ162.07,156.88,148.59,146.29,138.33,137.08,136.90,130.44,130.23,129.11,128.90,125.52,121.51,120.91,120.33,119.02,115.49,115.08,114.86,112.21,65.01,48.39,45.82,30.70,21.85,20.84.HRMS(ESI):calcd for C 32 H 32 ClN 5 O 3 [M+H] + 570.2259,found 570.2272.HPLC(80:20methanol:water with 1‰TFA):t R ＝7.92min,97.6％.
Example 51:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((((2-chloro- [1,1' -biphenyl ] -4-yl ] methyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 51)
(1) Preparation of 2-chloro- [1,1' -biphenyl ] -4-carboxylic acid
In 4-bromo-3-chlorobenzoic acid (3 g), phenylboronic acid (2 eq), pd (PPh 3 ) 4 (0.1 eq) toluene (30 ml) and Cs was added to the solution 2 CO 3 (2 eq). Heating and refluxing overnight under the protection of nitrogen, cooling the reaction liquid to room temperature after the reaction is completed, filtering, and preparing sand from the filtrate and performing column chromatography (PE/EA=80:1 to 50:1 v/v) t to obtain a white solid with the yield of 87%. 1 H NMR(300MHz,Chloroform-d)δ8.03(d,J＝1.5Hz,1H),7.91(dd,J＝7.5,1.5Hz,1H),7.60(d,J＝7.5Hz,1H),7.46(d,J＝4.6Hz,4H),7.38–7.33(m,1H).ESI-MS m/z:231.0[M-H] - .
(2) Preparation of 4- (bromomethyl) -2-chloro-1, 1' -biphenyl
The above 2-chloro- [1,1' -biphenyl]Preparation of 4-Carboxylic acid (2 g) after dissolution in THF (50 ml), BH under ice-bath conditions 3 THF (1M) was added slowly, followed by heating at 40℃overnight. After the reaction was completed, the reaction was quenched by addition of methanol, and then the solvent was suspended to obtain a white solid. The above white solid (1.5 g) was dissolved in THF (30 ml) and BBr was slowly added under ice bath conditions 3 (2 eq). The reaction solution was slowly warmed to room temperature, and after the reaction was completed, column chromatography was performed (PE/ea=80:1 to 50:1 v/v) to obtain a colorless liquid, yield was 57%, 1 H NMR(300MHz,Chloroform-d)δ7.52(q,J＝1.1Hz,1H),7.51–7.42(m,5H),7.39–7.34(m,1H),7.21(dq,J＝7.5,1.1Hz,1H),4.44(s,2H).ESI-MS m/z:281.1[M+H] + .
(3) Preparation of the title compound
The product is prepared from 3-bromotoluene and 4-chloro-o-methyl3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1, 1' -biphenyl was prepared according to example 1 in the form of a white solid with 37% yield, melting point: 157-158 ℃. 1 HNMR(300MHz,d6-DMSO)δ7.35(d,J＝34.8Hz,9H),7.11–6.92(m,2H),6.85–6.46(m,6H),4.80(s,2H),4.43(s,2H),3.83(s,2H),2.24(s,6H),2.16(s,5H). 13 C NMR(75MHz,DMSO)δ161.97,156.88,148.56,141.32,138.98,138.50,138.42,136.95,131.93,131.67,130.71,129.66,128.64,128.17,126.00,125.51,124.51,120.88,119.26,115.35,115.07,113.67,112.07,64.92,55.32,45.77,30.54,21.86,20.87.HRMS(ESI):calcd for C 36 H 34 Cl 2 N 2 O 3 [M+H] + 613.2032,found 613.2025.HPLC(80:20methanol:water with 1‰TFA):t R ＝10.2min,98.5％.
Example 52:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chloro-4-ethynylbenzyl) (m-tolyl) amino) -1H-pyrrole-2-carboxylic acid (compound 52)
(1) Preparation of (3-chloro-4- ((trimethylsilyl) ethynyl) phenyl) methanol
4-bromo-3-chlorobenzoic acid (4 g) was dissolved in THF (50 ml) and BH was slowly added under ice-bath conditions 3 THF, followed by heating overnight at 40 ℃. After the reaction was completed, the reaction was quenched by addition of methanol, followed by suspension to give a white solid. After the above white solid (3.5 g) was dissolved in triethylamine, pd (OAc) 2 (0.1 eq), cuI (0.1 eq) and trimethylsilyyne (2 eq) were added. Heating at 70deg.C for 30 hr under nitrogen protection. After the reaction was completed, it was cooled to room temperature, and sand-making and column chromatography (PE/ea=50:1 to 20:1 v/v) gave a yellow oil in 65% yield, 1 H NMR(300MHz,Chloroform-d)δ7.48(d,J＝7.5Hz,1H),7.35(s,1H),7.21(d,J＝8.8Hz,1H),4.74(s,2H),0.25(s,9H).ESI-MS m/z:237.1[M-H] - .
(2) Preparation of 4- (bromomethyl) -2-chloro-1-ethynyl benzene
The (3-chloro-4- "(trimethylsilyl) ethynyl) phenyl) methanol (1.5 g) was dissolved in THF (30 ml), TBAF-THF (1M) was added, the reaction mixture was stirred at room temperature for 30 minutes, and after completion of the reaction, 100ml of H was added 2 O, etOAc extracting (3 times), mixing the organic phases, drying with saturated saline, drying with sodium sulfate, and suspending to obtain brown oily substance, dissolving the brown oily substance in THF, adding BBr 3 (2 eq), stirring for 30 minutes at room temperature, sand making and column chromatography to obtain colorless liquid, the yield is 46%, 1 HNMR(300MHz,Chloroform-d)δ7.54–7.41(m,2H),7.24(dt,J＝7.5,1.3Hz,1H),4.40(s,2H),3.31(s,1H).ESI-MS m/z:229.0[M+H] + .
(3) Preparation of the title compound
This was prepared from 3-bromotoluene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1-ethynylbenzene as in example 1, as a white solid with 67% yield, melting point: 147-148 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.32(s,1H),7.50(d,J＝8.0Hz,1H),7.40(s,1H),7.21(d,J＝8.0Hz,1H),7.03(s,1H),6.97(t,J＝7.7Hz,1H),6.74(s,2H),6.71(s,1H),6.64–6.49(m,3H),4.76(s,2H),4.54(s,1H),4.48–4.39(m,2H),3.82(t,J＝6.0Hz,2H),2.26(s,6H),2.15(s,5H). 13 C NMR(75MHz,d6-DMSO)δ161.02,151.69,143.53,137.31,134.27,132.51,131.76,129.43,126.97,124.29,122.52,121.75,120.45,119.94,115.35,114.60,111.23,110.86,109.60,107.80,77.42,75.52,72.78,72.36,71.94,59.39,51.53,41.47,25.80,17.03,16.26.HRMS(ESI):calcd for C 32 H 30 Cl 2 N 2 O 3 [M+H] + 561.1700,found 561.1712.HPLC(80:20methanol:water with 1‰TFA):t R ＝15.9min,98.7％.
Example 53:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chloro-4-ethynylbenzyl) (3, 4-dimethylphenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 53)
The product is prepared from 3,4-Dimethyl bromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1-ethynylbenzene were prepared according to example 1 as yellow solid, 47% yield, melting point: 156-157 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.44(d,J＝7.9Hz,1H),7.34(s,1H),7.13(d,J＝8.0Hz,1H),6.94–6.85(m,2H),6.69(s,2H),6.63(s,3H),4.68(s,2H),4.48(t,J＝6.3Hz,2H),3.83(t,J＝5.4Hz,2H),3.38(s,1H),2.35(s,6H),2.26(d,J＝6.1Hz,2H),2.19(s,3H),2.18(s,3H).HRMS(ESI):calcd for C 33 H 32 Cl 2 N 2 O 3 [M+Na] + 597.1683,found 597.1688.HPLC(80:20methanol:water with1‰TFA):t R ＝14.6min,97.2％.
Example 54:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chloro-4-ethynylbenzyl) (3-chlorophenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 54)
This was prepared from 3-chlorobromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1-ethynylbenzene as per example 1, green solid, yield 63%, melting point: 156-157 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.41(s,1H),7.51(d,J＝7.9Hz,1H),7.40(d,J＝1.5Hz,1H),7.21(dd,J＝8.1,1.7Hz,1H),7.15–7.05(m,2H),6.77(d,J＝2.1Hz,1H),6.73(s,2H),6.72–6.64(m,3H),4.78(s,2H),4.54(s,1H),4.45(t,J＝6.5Hz,2H),3.82(t,J＝5.9Hz,2H),2.26(s,6H),2.15(p,J＝6.3Hz,2H). 13 C NMR(75MHz,d6-DMSO)δ161.89,156.87,149.95,142.31,136.93,135.51,134.43,134.18,130.88,129.46,127.71,125.70,125.58,121.37,120.29,117.52,115.01,114.49,113.43,112.64,86.25,80.39,64.91,55.61,45.93,30.52,20.87,14.41.HRMS(ESI):calcd for C 31 H 27 Cl 3 N 2 O 3 [M+H] + 583.1131,found 583.1136.HPLC(80:20methanol:water with1‰TFA):t R ＝15.7min,96.8％.
Example 55:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chloro-4-ethynylbenzyl) (3-fluorophenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 55)
This was prepared from 3-fluorobromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1-ethynylbenzene as in example 1, as a pink solid, 65% yield, melting point: 144-145 ℃. 1 H NMR(300MHz,d 6 -DMSO)δ12.40(s,1H),7.51(d,J＝8.0Hz,1H),7.40(d,J＝1.7Hz,1H),7.21(dd,J＝8.1,1.7Hz,1H),7.15–7.04(m,2H),6.77(d,J＝2.2Hz,1H),6.74(s,2H),6.59–6.42(m,3H),4.78(s,2H),4.54(s,1H),4.45(t,J＝6.5Hz,2H),3.83(t,J＝6.0Hz,2H),2.26(s,6H),2.16(q,J＝6.3Hz,2H).HRMS(ESI):calcd for C 31 H 27 Cl 2 FN 2 O 3 [M+H] + 565.1454,found 565.1461.HPLC(80:20methanol:water with 1‰TFA):t R ＝10.6min,97.2％.
Example 56:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chloro-4-ethynylbenzyl) (3-methoxyphenyl) amino) -1H-pyrrole-2-carboxylic acid (compound 56)
This was prepared from 3-methoxybromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1-ethynylbenzene as in example 1, as a white solid with a yield of 49%, melting point: 119-120 ℃. 1 H NMR(300MHz,d6-DMSO)δ12.34(s,1H),7.51(d,J＝8.0Hz,1H),7.40(d,J＝1.6Hz,1H),7.21(dd,J＝8.1,1.7Hz,1H),7.07(d,J＝2.2Hz,1H),6.99(t,J＝8.1Hz,1H),6.78–6.70(m,3H),6.40–6.25(m,3H),4.76(s,2H),4.53(s,1H),4.43(t,J＝6.4Hz,2H),3.83(t,J＝6.0Hz,2H),3.63(s,3H),2.26(s,6H),2.14(t,J＝6.2Hz,2H).HRMS(ESI):calcd for C 32 H 30 Cl 2 N 2 O 4 [M+H] + 577.1628,found 577.1661.HPLC(80:20methanol:water with 1‰TFA):t R ＝11.6min,96.7％.
Example 57:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((4-chloro-3-methylphenyl) (3-chloro-4-ethynylbenzyl) amino) -1H-pyrrole-2-carboxylic acid (compound 57)
The product was prepared from 3-methyl-4-chlorobromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1-ethynylbenzene according to example 1 as a yellow solid with a yield of 54%, melting point: 152-153 ℃. 1 H NMR(300MHz,CDCl 3 )δ7.46(d,J＝7.9Hz,1H),7.10(d,J＝7.9Hz,1H),7.00–6.90(m,2H),6.78(dd,J＝17.2,2.3Hz,2H),6.67–6.55(m,3H),4.66(s,2H),4.51(t,J＝6.4Hz,2H),3.83(t,J＝4.9Hz,2H),3.39(s,1H),2.35(s,6H),2.28(s,5H).HRMS(ESI):calcd for C 32 H 29 Cl 3 N 2 O 3 [M+Na] + 617.1141,found 617.1165.HPLC(80:20methanol:water with 1‰TFA):t R ＝12.5min,98.1％.
Example 58:
1- (3- (4-chloro-3, 5-dimethylphenoxy) propyl) -4- ((3-chloro-4-ethynylbenzyl) (m-tolyl) amino) -3, 5-dimethyl-1H-pyrrole-2-carboxylic acid (compound 58)
This was prepared from 3-methyl-4-chlorobromobenzene, 4-chloro-3, 5-dimethylphenol, 1, 3-dibromopropane, 4-nitro-1H-pyrrole-3, 5-dimethyl-2-carboxylic acid ethyl ester, 4- (bromomethyl) -2-chloro-1-ethynylbenzene as in example 1 as a yellow solid with 37% yield and melting point 168-169 ℃. 1 HNMR(300MHz,Chloroform-d)δ7.27–7.19(m,3H),7.12(d,J＝3.3Hz,1H),7.02–6.90(m,2H),6.76(s,1H),6.63(s,2H),4.68(s,2H),4.51(t,J＝6.2Hz,2H),3.83(t,J＝5.3Hz,2H),2.35(s,6H),2.28(s,6H),1.32(s,3H),0.98–0.86(m,2H).HRMS(ESI):calcd for C 34 H 34 Cl 2 N 2 O 3 [M+Na] + 611.1844,found611.1850.HPLC(80:20methanol:waterwith1‰TFA):t R ＝13.7min,98.5％.
Activity test examples:
1. test of Compounds for inhibitory Activity of Mcl-1, bcl-2 and Bfl-1 Using fluorescence bias method test (FP experiment)
The inhibition activity of a target compound on Mcl-1, bcl-2 and Bfl-1 is tested by adopting an FP method, the final concentration of the protein is 20nM for Mcl-1 protein, and the selected probe is FITC-Bak-BH3 and the concentration of the probe is 10nM; for Bcl-2 protein, the final protein concentration is 20nM, the probe is Fam-Bid, and the final probe concentration is 0.66nM; for Bfl-1 protein, the final protein concentration was 13.3nM, the probe was Fam-Bid, and the final probe concentration was 0.33nM. The assay used 384 Kong Heiban (model Corning 3676), the final volume of the assay was selected to be 40. Mu.L and the compound tested and FITC-9merNrf2 polypeptide fluorescent probe were dissolved in DMSO (10 mmol stock solution) for use. The compounds were diluted 11 concentration gradients with HEPES buffer, 20. Mu.L of diluted compound was added to each well plate, followed by 10. Mu.L of 10nM probe diluted with buffer and 10. Mu.L of 12nM protein. Two duplicate wells were set for each compound concentration, and a blank (10. Mu.L probe+40. Mu.L buffer, denoted Pmin) and a negative (10. Mu.L probe+10. Mu.L protein+20. Mu.L buffer, denoted Pmax) were set up for the experiment. After incubation on a shaker for 30min with shaking at room temperature, the plate was swept with SpectraMaxMulti-ModeMicroplateReader (MolecularDevices), excitation wavelength set to 485nm, emission wavelength set to 535nm and test well reading recorded as Pobs. The inhibition rate calculation formula is: inhibition ratio (%) = [1- (Pobs-Pmin)/(Pmax-Pmin) ]X 100% IC was calculated using graphpadprism6.0 software 50 . Ki is calculated according to the following formula:
K i ＝[I] 50 /([L] 50 /K d +[P] 0 /K d +1)
wherein [ I ]50 represents half inhibition concentration of the compound, [ L ]50 represents probe concentration, [ P ]0 represents protein concentration, kd represents dissociation constant of the protein and the probe.
The test results indicate that most of the compounds have Mcl-1 nanomolar inhibitory activity. The results are shown in Table 1.
TABLE 1
2. Bcl-2 family subtype selectivity of representative compounds
The inhibition results of compounds at 10. Mu.M concentration on Bcl-2, bfl-1, bcl-xL indicated good subtype selectivity for active compounds compound 41, compound 51 and compound 52. The results are shown in Table 2.
TABLE 2 evaluation of Bcl-2 subtype selectivity of representative Compounds
The results show that all of our compounds had no significant inhibitory activity against 3 Bcl-2 subtypes Bcl-2, bcl-xL and Bfl-1 at a concentration of 10. Mu.M. Thereafter, we selected compound 52 and evaluated the affinity selectivity of the compound for Bcl-2 family proteins using Bcl-2scan technology from discovery x, and the results are shown in figure 1. The results in FIG. 1 show that the Mcl-1 affinity of compound 52 achieves a low nanomolar (K d =0.063 nM), selectivity for Bcl-2 other subtypes Bcl-2, bcl-xL, bcl-w, and Bcl-2A1 is over 10 6 Multiple (K) d ≥10000nM)。
3. Evaluation of in vitro antitumor Activity of representative Compounds
After confirming the target activity and subtype selectivity of representative compounds, we then evaluated the in vitro antitumor activity of compound 41, compound 51, compound 52. The in vitro anti-tumor activity of the compounds against the Mcl-1 sensitive cell line human myeloma cell line H929, the human leukemia cell line MV4-11, the human breast cancer cell line SX-BR3 and the human non-small cell lung cancer cell line NCI-H23 was evaluated using the thiazole blue (MTT) method with Mc-1 inhibitor A1210477 reported by Ebolv company as a positive control; the antiproliferative activity of the compounds against Mcl-1 insensitive cell line human leukemia cell line K562 was also evaluated using the MTT method and the results are shown in table 3.
Table 3 evaluation of in vitro antitumor Activity of representative Compounds
MTT assay showed that all compounds had good antiproliferative activity against H929 cell lines and compound 52 showed antiproliferative activity superior to that of positive drug a 1210477; for MV4-11 cells, the activity of all compounds was reduced, but compound 52 still maintained low micromolar activity; the anti-proliferative activity of all compounds SX-BR3, NCI-H23 was significantly reduced, consistent with the reduction in anti-proliferative activity against solid tumors exhibited by Mcl-1 inhibitors reported previously; all compounds were poorly active against Mcl-1 insensitive strain K562, indicating that the compounds have good cell selectivity.
4. Evaluation of antitumor Activity of Compound 52 in MV4-11 nude mouse transplantation tumor model
We constructed MV4-11balb/c nude mouse model to evaluate the in vivo antitumor activity of compound 52, and selected the targeted Mcl-1 clinical candidate compound AZD5991 reported by the company Aschikang as a positive control, and compound 52 was administered in both gastric lavage and intraperitoneal injection, with AZD5991 administered intraperitoneally, once every two days for 14 days. As a result, as shown in fig. 2, compound 52 was able to significantly inhibit tumor growth in nude mice, with T/C up to 8.2% in the orally administered group (80 mg/kg); the T/C of the compound 52 intraperitoneal administration group (20 mg/kg) reached 1.0%, which is superior to the activity of the AZD5991 intraperitoneal administration group (20 mg/kg) (T/c=3.3%). Compound 52 did not cause significant changes in nude mice body weight and heart, liver, kidney weight during administration.
Claims (9)
1. A4-amino-1H-pyrrole compound has the following structural general formula:
R 1 and R is 2 Selected from the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, halogen, hydroxy, mercapto, alkoxy, and alkylthio; r is R 3 And R is 4 Each independently selected from a hydrogen atom, a substituted or unsubstituted C1-C5 alkyl group.
2. The 4-amino-1H-pyrrole compound according to claim 1, wherein the alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl substituted with one or more substituents selected from alkyl, halogen, hydroxy, mercapto, amino, oxo, carboxyl, nitro, cyano, alkoxy, alkylthio, cycloalkyl, heterocyclyl, aryl and heteroaryl.
3. The 4-amino-1H-pyrrole compound according to claim 2, R 1 Selected from phenyl, 2-pyridyl, 1-naphthyl, 2-methylphenyl, 3-methylphenyl, 4-ethylphenyl, 4-cyclopropylphenyl, 4-isopropylphenyl, 4-t-butylphenyl, 3-ethylphenyl, 3-isopropylphenyl, 3-t-butylphenyl, 3-chlorophenyl, 3-fluorophenyl, 3-acetylphenyl, 3-nitrophenyl, 3-cyanophenyl, 3-methoxyphenyl, 3-hydroxyphenyl, 3- (1-hydroxyethyl) phenyl, 3, 4-dimethylphenyl and 3-methyl-4-chlorophenyl.
4. The 4-amino-1H-pyrrole compound according to claim 2, R 2 Selected from 3-thienyl, 1-naphthyl, 2-naphthyl, phenyl, 2-chlorophenyl, 2-methylphenyl, 3-chloromethyl, 3-methylphenyl, 4-chlorophenyl, 4-methylphenyl, 3-bromophenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-trifluoromethylphenyl, 3-trifluoromethoxyphenyl, 3-aldehydylphenyl, 4-isopropylphenyl, 4-tert-butylphenyl, 4-biphenyl, 1-phenyl-1H-1, 2, 3-triazol-4-yl4-ethynylphenyl, 4-propynylphenyl, 3,4 dimethylphenyl, 3-bromo-4 methylphenyl, 3, 4-dichlorophenyl, 3,5 dimethylphenyl, 2, 3-difluorophenyl, 2, 4-dichlorophenyl, 2, 6-dichlorophenyl, 3-chloro-4 phenylphenyl and 3-chloro-4-ethynylphenyl.
5. A process for preparing a 4-amino-1H-pyrrole compound according to claim 1, R 1 Definition of R is as defined in claim 1 2 =3-chlorophenyl group, R 3 、R 4 Is a hydrogen atom, and comprises the following synthetic routes and steps:
raw materials I-2 and 1, 3-dibromopropane are in acetonitrile, potassium carbonate is taken as an acid binding agent, nucleophilic substitution reaction is carried out to obtain an intermediate I-3, the intermediate I-1 and the intermediate I-3 are under the action of cesium carbonate to obtain an intermediate I-4 in DMF, the intermediate I-4 is subjected to nucleophilic substitution reaction to obtain I-5, I-5-and different bromoband aromatic hydrocarbons in Pd through stannous chloride reduction 2 (dba) 3 Under the catalysis of BINAP, cs is used 2 CO 3 As alkali, obtaining an intermediate I-6, I-6 and 3-chlorobenzyl through Buchward-Hartwig reaction, taking NaH as an acid binding agent, carrying out nucleophilic substitution reaction with 3-chlorobenzyl to obtain an intermediate I-7, then hydrolyzing the intermediate I-7 to obtain a target product and obtaining an intermediate I-8,I-8 through H 2 And (3) debenzylating Pt/C to obtain a target product a, and reducing the target product substituted by the 3-acetylphenyl R1 by sodium borohydride to obtain a target product b.
6. A process for preparing a 4-amino-1H-pyrrole compound according to claim 1, R 1 =3-methylphenyl, R 2 Defined as follows, R 3 、R 4 Is a hydrogen atom, and comprises the following synthetic routes and steps:
R 2 =3-thienyl, 1-naphthyl, 2-naphthyl, phenyl, 2-chlorophenyl, 2-methylphenyl, 3-chloromethyl, 3-methylphenyl, 4-chlorophenyl, 4-methylphenyl, 3-bromophenyl, 3-methoxybenzene A group, 3-fluorophenyl, 3-trifluoromethylphenyl, 3-trifluoromethoxyphenyl, 3-aldehydylphenyl, 4-isopropylphenyl, 4-tert-butylphenyl, 4-biphenylyl, 1-phenyl-1H-1, 2, 3-triazol-4-yl, 4-ethynylphenyl, 4-propynylphenyl, 3,4 dimethylphenyl, 3-bromo-4-methylphenyl, 3, 4-dichlorophenyl, 3,5 dimethylphenyl, 2, 3-difluorophenyl, 2, 4-dichlorophenyl or 2, 6-dichlorophenyl group;
NaH is used as an acid binding agent, intermediate I-9 and bromobenzyl with different substitutions are subjected to nucleophilic substitution reaction to obtain intermediate II-1, II-2, the intermediate II-1 and II-2 are hydrolyzed by NaOH to obtain a target product a, one of the intermediate II-2 and azidobenzene are subjected to click reaction under the action of DIPEA and CuI to obtain intermediate II-3, and the intermediate II-3 is hydrolyzed by NaOH to obtain a target product b.
7. A process for preparing a 4-amino-1H-pyrrole compound according to claim 1, R 1 3-methylphenyl, 3-chloromethyl, 3-fluoromethylphenyl, 3-methoxyphenyl, 3, 4-dimethylphenyl, 3-chloro-4-methylphenyl, R 2 =3-chloro-4-phenylphenyl, 3-chloro-4-ethynylphenyl, R 3 、R 4 Is hydrogen atom or methyl, comprising the following synthetic route and steps:
starting material III-1 with phenylboronic acid in Pd (PPh 3 ) 4 Under the catalysis, obtaining an intermediate III-2 through Suzuki reaction, obtaining benzyl alcohol III-3 through borane reduction of III-2, and obtaining bromobenzyl III-4 through reaction of III-3 and phosphorus tribromide; reducing III-1 with borane to obtain benzyl alcohol III-5, III-5 and trimethylsilylacetylene in PdCl 2 (PPh 3 ) 2 Under the catalysis of cuprous iodide, introducing alkynyl through Sonogashira coupling to obtain intermediate III-6, deprotecting III-6 with TBAF to obtain III-7, and mixing with PBr 3 Obtaining bromobenzyl III-8 by reaction, obtaining intermediate III-10 by nucleophilic substitution reaction of intermediate III-9 and III-4 and III-8 under the action of NaH, and obtaining target product by NaOH hydrolysis of intermediate III-10.
8. Use of a 4-amino-1H-pyrrole compound according to any one of claims 1 to 4 for the preparation of a Mcl-1 inhibitor medicament.
9. Use of the 4-amino-1H-pyrrole compound according to any one of claims 1 to 4 for the preparation of antitumor drugs.
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