阅读说明：本技术 噻吩并吡咯酰胺类衍生物及其在抗肿瘤药物中的应用 (Thienopyrrole amide derivatives and application thereof in antitumor drugs ) 是由 杨金飞 于 2021-07-29 设计创作，主要内容包括：本发明属于药物化学技术领域,尤其涉及噻吩并吡咯酰胺类衍生物和制备方法,及其作为MEK抑制剂在抗肿瘤药物中的应用。本发明的提供一种通式所示的噻吩并吡咯酰胺类衍生物,及其几何异构体或其药学上可接受的盐、水合物、溶剂化物或前药；本发明所述MEK抑制剂具有特异性和有效性,具有更为广阔的发展前景。本发明通过实验结果显示,本课题组合成的噻吩并吡咯酰胺类衍生物具有开发靶向抗肿瘤药物的前景。(The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a thienopyrrole amide derivative, a preparation method thereof and application thereof as an MEK inhibitor in antitumor drugs. The invention provides a thienopyrrole amide derivative shown in a general formula, and a geometric isomer thereof or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof; the MEK inhibitor provided by the invention has specificity and effectiveness, and has a wider development prospect. The experimental results show that the thienopyrrole amide derivatives synthesized by the subject combination have the prospect of developing targeted antitumor drugs.)

1. The thienopyrrole amide derivative is characterized by having a structural formula as follows:

wherein, R is₁Selected from hydrogen, halogen, C₁-C₆Alkyl, alkenyl or alkynyl;

x or Y is selected from hydrogen, halogen, C₁-C₆An alkyl group.

2. Thienopyrrole amide derivatives according to claim 1, selected from:

3. the thienopyrrole amide derivatives as claimed in any one of claims 1 to 2 are useful as antitumor agents.

Technical Field

The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a thienopyrrole amide derivative, a preparation method thereof and application thereof as an MEK inhibitor in antitumor drugs.

Background

Malignant tumors seriously affect the life span of human beings, and the incidence and mortality rate are on a worldwide increasing trend year by year. About 1810 thousands of new malignant tumor patients are globally present in 2018. However, in China, 410 million malignant tumor patients are newly increased every year, and 260 million people die from malignant tumor. Target therapy, which is directed to the type of gene mutation, has become a major trend in the treatment of malignant tumors. The RAS/RAF/MEK/ERK signaling pathway plays an important role in tumorigenesis and progression, regulating cell proliferation and apoptosis. Make internal disorder or usurp found that one third of the malignant tumors had upregulated RAS/RAF/MEK/ERK pathways. RAS mutated tumors account for 30% of all tumors and RAF mutated tumors account for 7% of all tumors. MEK kinases are located downstream of RAF and comprise 7 kinases (MEK1, MEK2, MEK 3-7). MEK1 and MEK2 are closely related, dual specificity tyrosine/threonine protein kinases. MEK kinases selectively phosphorylate tyrosine/threonine and tyrosine residues within their specific MAP kinase substrate active loops. MEK comprises an amino terminus, a catalytic domain, and a carboxy terminus. The catalytic domain is highly homologous, while the amino and carboxy termini are diverse. ERK1 and ERK2 are the only substrates of MEK1/2 known at present, and thus MEK1/2 is an attractive target for tumor therapy.

In recent years, there has been considerable progress in the development of MEK inhibitors and their use in the treatment of tumors. CI-1040 is the first MEK inhibitor to enter clinical trials, developed and advanced by the company Pereri/Washanburt. The compound is a novel benzohydroxamate compound, and an MEK inhibitor derived from anthranilic acid serving as a precursor of the benzohydroxamate compound. However, due to poor solubility, rapid clearance and poor exposure to blood, CI-1040 did not reach ideal levels, leading to failure of the two phase II studies make internal disorder or usurp in 2003 due to insufficient antitumor activity. Efforts to optimize the hydroxamate side chain of CI-1040 to improve solubility and exposure to oral administration, then, pfeir/warna-lambert discovered PD-0325901 by pfeiffer/warna-lambert. However, in clinical trials, patients were observed that the compound could cross the blood brain barrier and produce neurological side effects. Next, a series of structural optimization works of the compound were performed, and cobicistinib, which was approved by FDA and marketed in 2015, was found, but the pharmaceutical effect of the compound was not prominent. Meanwhile, on the basis of PD-0325901, the arreisan organism discovers a new series of benzimidazole compound, namely, erlotinib, wherein 4-N is used for substituting 4-F, so that H receptors are reserved, the penetration of a blood brain barrier is avoided, and unfortunately, the pharmaceutical activity is reduced. Another compound, beninib, was FDA approved in 2018 and has the same core structure as AZD 244. However, this compound was found to be able to cross the blood brain barrier, leading to central nervous system toxicity. For a long time, the field of drug synthesis research has been concerned with the development of novel, scaffold, high-efficiency, non-toxic and non-toxic MEK inhibitors.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a novel thienopyrrole amide derivative; and a preparation method of the derivative and application of the derivative as an MEK target inhibitor in antitumor drugs.

In order to achieve the purpose, the invention adopts the technical scheme that: the invention provides a thienopyrrole amide derivative shown in a general formula (I), and a geometric isomer thereof or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof;

the R is₁Selected from hydrogen, halogen, C₁-C₆Alkyl, alkenyl or alkynyl.

X or Y is selected from hydrogen, halogen, C₁-C₆An alkyl group.

The N-cyclopropyl methoxy imidazole amide derivatives shown in the general formula (I) are selected from the following compounds:

the compound of formula I can be synthesized according to the following synthetic route method, substituted 4H-thiophene [3,2-b ] pyrrole-5-carboxylic acid ethyl ester is used as a raw material, nucleophilic substitution reaction is firstly carried out on the substituted 4H-thiophene [3,2-b ] pyrrole-5-carboxylic acid ethyl ester and substituted benzyl bromide to obtain an intermediate 2, hydrolysis reaction is carried out under alkaline condition to obtain an intermediate 3, and condensation reaction is finally carried out on the intermediate 3 and 3-amino-1, 2-propylene glycol under condensing agent condition to obtain a target compound.

Reagents and conditions: (a) k2CO3, DMF,80 ℃; (b)2N NaOH, MeOH/H2O; (c) EDCI, HOBt, DIEA, DMF, rt.

The thienopyrrole amide derivatives can be used as antitumor drugs, and particularly as MEK target inhibitors.

The invention has obvious technical effect.

The research and development of MEK target inhibitor drugs are one of the main research hotspots of the current anti-tumor drugs. The discovery of such drugs has brought new hopes for the treatment of patients with tumors. Has higher specificity and effectiveness and wider development prospect. The experimental result shows that the N-cyclopropyl methoxyimidazole amide derivatives synthesized by the composition have the prospect of developing targeted antitumor drugs.

Detailed Description

The following examples are intended to illustrate but not limit the scope of the invention. The starting materials are generally available from commercial sources or may be prepared using methods well known to those skilled in the art or in accordance with the methods described herein. The reagents used are, without particular reference, analytically or chemically pure. The nuclear magnetic resonance hydrogen spectrum of the compound is measured by BrukeraRx-400, and the mass spectrum is measured by Agilent 1100 LC/MS; all reagents used were analytically or chemically pure.

Example 1.

Step 1 Synthesis of intermediate 2

Sodium hydrogen carbonate (0.51g,12.81mmol) was dissolved in DMF and 4H-thiophene [3,2-b ] was slowly added dropwise]After the dropwise addition of a solution of pyrrole-5-carboxylic acid ethyl ester (1.00g,5.12mmol) in DMF, the reaction was continued at room temperature for 10min, then benzyl bromide (1.05g,6.15mmol) was added, after 4 hours of reaction, TLC detection of completion of the reaction was carried out, 20mL of a 10% ammonium chloride solution was added to the reaction solution, extraction was carried out with ethyl acetate, the organic layer was washed with saturated saline, and Na was added₂SO₄Dry overnight. The drying agent was filtered off, the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography to give 1.21g of a pale yellow oil with a yield of 82.78%.¹H-NMR(400MHz,CDCl₃)δ7.35–7.04(m,7 H),6.84(s,1H),5.77(s,2H),4.30(q,J＝7.0Hz,2H),1.34(t,J＝7.1Hz,3H).

Step 2 Synthesis of intermediate 3

Dissolving the intermediate 2(1.00g,6.64mmol) in 20mL ethanol, adding 10mL 2N sodium hydroxide solution, stirring at room temperature for 2h, detecting by TLC, removing ethanol under reduced pressure, adding water, adjusting pH to 4-5 with 3N hydrochloric acid, precipitating a large amount of solid, and filtering to obtain white solid 0.82g with yield of 90.4%.

Step 3 Synthesis of example 1

Intermediate 3(0.50g,1.94mmol) was dissolved in 20mL DMF and EDCI (0.41g, 2.14mmol) and HOBt (0.29g,2.14mmol) were added. After 1h reaction at room temperature, 3-amino-1, 2-propanediol (0.35g,3.89mmol) and DIEA (0.62g,4.86mmol) were added and the reaction was allowed to proceed at 80 ℃ for 12 h. TLC detection reaction is completed, the reaction temperature is reduced to room temperature, the reaction liquid is poured into 100mL of water, solid is separated out, filtration and drying are carried out, and the crude product is purified by silica gel column chromatography to obtain 0.35g of white solid with the yield of 54.5%.¹H-NMR(400 MHz,DMSO-d₆)δ8.98(s,1H),7.33–7.07(m,7H),7,06(s,1H),5.77(s,2H),5.38 (s,1H),4.91(s,1H),4.24–4.21(m,1H),3.59–3.53(m,2H),3.29–3.26(m,1H), 3.08–3.04(m,1H).ESI-MS m/z:331.1[M+H]⁺.

Examples 2-11 were prepared according to the procedure for example 1, starting with substituted ethyl 4H-thieno [3,2-b ] pyrrole-5-carboxylate and various substituted benzyl bromides, respectively, via three steps of alkylation, hydrolysis and condensation.

Example 2.

¹H-NMR(400MHz,DMSO-d₆)δ8.96(s,1H),7.31–7.11(m,7H),5.78(s,2H), 5.39(s,1H),4.90(s,1H),4.25–4.22(m,1H),3.58–3.53(m,2H),3.29–3.25(m, 1H),3.07–3.03(m,1H).ESI-MS m/z:349.1[M+H]⁺.

Example 3.

¹H-NMR(400MHz,DMSO-d₆)δ8.96(s,1H),7.31–7.10(m,7H),5.79(s,2H), 5.36(s,1H),4.92(s,1H),4.25–4.22(m,1H),3.60–3.54(m,2H),3.28–3.25(m, 1H),3.08–3.05(m,1H).ESI-MS m/z:365.1[M+H]⁺.

Example 4.

¹H-NMR(400MHz,DMSO-d₆)δ8.96(s,1H),7.41(d,J＝5.4Hz,1H), 7.13–7.07(m,4H),7.02(d,J＝8.4Hz,2H),5.78(s,2H),5.35(s,1H),4.90(s,1H), 4.25–4.22(m,1H),3.59–3.54(m,2H),3.29–3.26(m,1H),3.09–3.05(m,1H), 2.19(s,3H).ESI-MS m/z:345.1[M+H]⁺.

Example 5.

¹H-NMR(400MHz,DMSO-d₆)δ8.99(s,1H),7.40(d,J＝5.0Hz,1H), 7.13–7.08(m,4H),6.86(d,J＝8.5Hz,2H),5.81(s,2H),5.35(s,1H),4.90(s,1H), 4.25–4.21(m,1H),3.80(s,3H),3.60–3.54(m,2H),3.30–3.26(m,1H),3.09– 3.04(m,1H).ESI-MS m/z:361.1[M+H]⁺.

Example 6.

¹H-NMR(400MHz,DMSO-d₆)δ8.94(s,1H),7.67(d,J＝2.0Hz,1H),7.40(d, J＝5.0Hz,1H),7.16–7.07(m,4H),5.75(s,2H),5.34(s,1H),4.90(s,1H),4.25– 4.22(m,1H),3.59–3.54(m,2H),3.29–3.25(m,1H),3.09–3.05(m,1H).ESI-MS m/z:399.0[M+H]⁺.

Example 7.

¹H-NMR(400MHz,DMSO-d₆)δ8.97(s,1H),7.15–7.11(m,5H),6.77(d, J＝5.4Hz,1H),5.81(s,2H),5.36(s,1H),4.92(s,1H),4.26–4.22(m,1H),3.59– 3.53(m,2H),3.29–3.24(m,1H),3.07–3.02(m,1H),2.34(s,3H).ESI-MS m/z: 363.1[M+H]⁺.

Example 8.

¹H-NMR(400MHz,DMSO-d₆)δ8.94(s,1H),7.31(d,J＝8.4Hz,2H), 7.14–7.10(m,3H),6.75(d,J＝5.4Hz,1H),5.80(s,2H),5.37(s,1H),4.90(s,1H), 4.26–4.22(m,1H),3.61–3.54(m,2H),3.28–3.25(m,1H),3.08–3.04(m,1H), 2.37(s,3H).ESI-MS m/z:379.1[M+H]⁺.

Example 9.

¹H-NMR(400MHz,DMSO-d₆)δ8.97(s,1H),7.13–7.08(m,3H),7.02(d, J＝8.4Hz,2H),6.75(d,J＝5.4Hz,1H),5.81(s,2H),5.36(s,1H),4.91(s,1H),4.24– 4.21(m,1H),3.59–3.54(m,2H),3.29–3.26(m,1H),3.09–3.05(m,1H),2.38(s, 3H)，2.20(s,3H).ESI-MS m/z:359.1[M+H]⁺.

Example 10.

¹H-NMR(400MHz,DMSO-d₆)δ8.98(s,1H),7.31(d,J＝5.4Hz,1H), 7.18–7.11(m,6H),5.36(s,1H),5.08–5.04(m,1H),4.89(s,1H),4.24–4.21(m, 1H),3.57–3.53(m,2H),3.29–3.25(m,1H),3.07–3.01(m,1H),1.64(d,J＝7.0Hz, 3H).ESI-MS m/z:363.1[M+H]⁺.

Example 11.

¹H-NMR(400MHz,DMSO-d₆)δ8.96(s,1H),7.35–7.30(m,3H),7.15–7.11 (m,4H),5.34(s,1H),5.07–5.03(m,1H),4.94(s,1H),4.25–4.21(m,1H),3.58– 3.52(m,2H),3.29–3.25(m,1H),3.06–3.02(m,1H),1.62(d,J＝7.1Hz, 3H).ESI-MS m/z:379.1[M+H]⁺.

The invention also relates to the pharmacological research of the partial product.

1. Inhibitory Activity of Compounds on MEK1 Using Z' -LYTE^TMKinase assay methods compounds were tested for their effect on MEK1 kinase activity. In 384-well plates, the reaction system is 50mM HEPES pH7.5, 10mM MgCl₂1mM EGTA, 0.01% BRIJ-35,0.2nM MEK1,114.8 nMRK2. Test compounds were diluted sequentially to 8 concentrations. After being mixed evenly, the mixture reacts for 10min under the condition of room temperature, then 45 mu MATP and 1.5 mu M Ser/Thr3 peptide are added to start the reaction, and the incubation is continued for 1h under the room temperature. And finally adding a stop solution to terminate the reaction. Finally, fluorescence value measurement was performed using a microplate reader.

Table 1 the compounds of the examples are active in inhibiting MEK 1.

Examples	IC50(μM)	Examples	IC50(μM)
				Example 1	0.42	Example 7	0.38
Example 2	0.19	Example 8	0.62
				Example 3	0.28	Example 9	1.12
Example 4	0.52	Example 10	0.58
				Example 5	0.65	Example 11	0.73
Example 6	0.11

The results show that the compounds of examples 1-11 of the present invention have significant inhibitory effect on MEK 1.

2. Cell proliferation inhibition assay (MTT assay): tumor cells A375 and HT-29 cells were selected, cultured to logarithmic growth phase, adherent cells were treated with trypsin digestion and collected in DMEM medium containing 10% fetal bovine serum. The cell suspension was centrifuged (1000 Xrpm) and the cells were diluted to 2.5-5.0X 10³Per mL, per wellAdding 2.0-3.0X 10³The cells were cultured at 37 ℃ for 24 hours. Adding 2 μ L of drug solution with different concentrations, culturing at 37 deg.C, adding 10 μ L of MTT [3- (4, 5-dimethylthiazole-2) -2, 5-diphenyl tetrazolium bromide salt at different time points]And (3) solution. Incubation at 37 ℃ for 4h, medium was discarded, 200 μ L DMSO was added per well to dissolve residual formazan crystals, and after 15min, absorbance was recorded at 490 nm.

TABLE 2 IC of Compounds 1-11 for tumor cell inhibitory Activity₅₀。