CN114773274A - Novel 4, 6-pyrimidine derivatives and application thereof in antitumor drugs - Google Patents

Novel 4, 6-pyrimidine derivatives and application thereof in antitumor drugs Download PDF

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CN114773274A
CN114773274A CN202210478645.0A CN202210478645A CN114773274A CN 114773274 A CN114773274 A CN 114773274A CN 202210478645 A CN202210478645 A CN 202210478645A CN 114773274 A CN114773274 A CN 114773274A
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王誉熹
张吉发
谭论
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West China Precision Medicine Industrial Technology Institute
West China Hospital of Sichuan University
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    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
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Abstract

The invention relates to a tubulin inhibitor and application thereof in a lung cancer medicament, and belongs to the technical field of antitumor science. The technical problem to be solved by the invention is to provide a compound of a tubulin inhibitor. The compound or the pharmaceutically acceptable salt thereof can be used as a tubulin inhibitor, has a certain anti-lung cancer activity and can effectively inhibit the growth of lung cancer cells.

Description

Novel 4, 6-pyrimidine derivatives and application thereof in antitumor drugs
Technical Field
The invention discloses a targeted tubulin inhibitor and application thereof in lung cancer treatment, belonging to the technical field of antitumor medicine.
Background
Lung cancer is one of the most common malignant tumors in the world, accounting for about 80% of all lung cancers, with about 75% of patients found in the middle and advanced stages with very low 5-year survival rate.
Microtubule-targeted drugs can affect the tubulin dynamics during cancer cell mitosis, preventing mitotic spindle-mediated correct chromosome segregation, leading to disruption of the cell cycle process and even cell death. Seven tubulin binding sites have been identified, including the taxane site, the vinca site, the colchicine site, the pironetite site, the maytansine site, the laulimolide/peloruside site and the gatobulin site. Among other advantages, Colchicine Binding Site Inhibitors (CBSI) have over-expression of tubulin isoforms effectively, overcoming resistance mediated by P-gp, MRP1 and MRP2, and damaging effects on tumor vasculature. Although showing better clinical effects for patients, the development of drug resistance and poor water solubility are the most major problems limiting the clinical use of CBSI. However, the current research on CBSI is mainly based on structural optimization of the reported inhibitors. Therefore, CBSI with a novel backbone was found to be an effective way to treat cancer and overcome multidrug resistant tumors.
Disclosure of Invention
The technical problem solved by the invention is to provide a novel compound serving as a colchicine binding site inhibitor.
A compound having the structural formula I:
Figure BSA0000272513520000011
wherein R is1Is composed of
Figure BSA0000272513520000012
Figure BSA0000272513520000013
The invention also provides application of the compound or the pharmaceutically acceptable salt thereof in preparing antitumor drugs.
Further, the anti-tumor therapeutic drug is preferably a lung cancer therapeutic drug.
Further, the lung cancer treatment drug is a tubulin colchicine binding site small molecule inhibitor, and the application of the lung cancer treatment drug is used for lung cancer related treatment.
The present invention also provides a pharmaceutical composition which is a preparation comprising an effective dose of the above-mentioned compound or a pharmaceutically acceptable salt thereof.
The compound or the pharmaceutically acceptable salt thereof prepared by the invention can be used as a tubulin colchicine binding site small molecule inhibitor, and has a relatively obvious lung cancer treatment effect.
Drawings
Fig. 1A is a photograph of colony formation six days after a549 cells in logarithmic growth phase were cultured in media containing different concentrations of compound 10.
FIG. 1B is a histogram showing the colony formation inhibition rate.
Figure 2A is a view of the interaction between compound 10 and tubulin (compound 10 is shown in yellow) and the corresponding structure rotated 90 ° counterclockwise along the X-axis.
FIG. 2B is a picture of A549 cells treated with 0.1% DMSO, Taxol (100nM), colchicine (100nM) and Compound 10(100nM) for 18 hours.
Fig. 3A is a picture of a549 cells cultured with 0.1% DMSO and different concentrations of compound 10 for 48 hours and analyzed by flow cytometry.
Figure 3B is a histogram showing the percentage of apoptotic index distribution treated with compound 10.
FIG. 3C is a photograph of Western blotting analysis of the effect of Compound 10 on apoptosis-related proteins.
Fig. 4A is a representative picture of a wound healing experiment.
Fig. 4B is a picture of wound closure rate after 12 hours of treatment with 0.1% DMSO and compound 10.
Fig. 4C is a representative picture of the Transwell migration test.
Figure 4D is a histogram of migrating cells after 12 hours of treatment with 0.1% DMSO and compound 10.
Detailed Description
The invention provides a compound shown as a formula I and a pharmaceutically acceptable salt thereof:
Figure BSA0000272513520000021
wherein R is1Is composed of
Figure BSA0000272513520000022
Figure BSA0000272513520000023
As a preferred embodiment, R is preferably1Is composed of
Figure BSA0000272513520000024
The following are some preferred structures of the compounds of the present invention.
Figure BSA0000272513520000031
The invention also provides pharmaceutically acceptable salts of the compounds described herein. The salt may be nitrate, hydrochloride, sulfate or phosphate, etc.
The invention also provides application of the compound or the pharmaceutically acceptable salt thereof in preparing a medicament for treating lung cancer. Further, the lung cancer therapeutic drug is preferably a microtubule small molecule inhibitor.
The present invention also provides a pharmaceutical composition which is a preparation comprising an effective dose of the above-mentioned compound or a pharmaceutically acceptable salt thereof. The compounds of the invention can be prepared in the following forms by methods known in the art: tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, finely divided powders or aerosols or sprays for inhalation, sterile aqueous or oily solutions or suspensions or sterile emulsions for parenteral (including intravenous, intramuscular or infusion). The active ingredient may be formulated in aqueous polyethylene glycol solutions, or liquid preparations may be prepared using water-propylene glycol solutions or sterile water as the solvent. Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents as desired. Aqueous suspensions for oral use can be made by dispersing the tiny and dispersed active ingredient in water together with natural synthetic gums, resins, methylcellulose, carboxymethylcellulose and other suspensions known in the pharmaceutical arts.
The pharmaceutical composition may be in unit dosage form. In these forms, the compositions are divided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation comprising discrete quantities of the preparation, such as tablets in boxes, capsules, and powders in vials or ampoules . The unit dosage form can also be a cachet or a tablet or it can be the appropriate number of all of these packaged forms.
The active ingredient of the pharmaceutical composition of the present invention may be only the compound of the present invention, or may be combined with other anti-lung cancer compounds as an active ingredient.
In the treatment of lung cancer, combination therapy can be achieved by administering the various therapeutic ingredients separately, simultaneously or sequentially. Such combinations employ the compounds of the invention in an effective dosage range and the other pharmaceutically active agents in a permitted dosage range.
The following examples are provided to further illustrate the embodiments of the present invention and are not intended to limit the invention to the embodiments described.
Example 1 Synthesis of Compounds 1-15.
Compounds 1-15 were synthesized using the following reaction formulas:
Figure BSA0000272513520000032
Reagents and conditions：(a)DIPEA，EtOH，4，6-dichloropyrimidine，80℃；(b)TFA，DMF，60℃
intermediate 1 a.
N- (3-aminophenyl) cyclopropylamide (0.8g, 4.54mmol) was dissolved in 15ml ethanol and 4, 6-dichloropyrimidine (1.0g, 6.71mmol) and DIPEA (1.5ml, 9.07mmol) were added. The reaction mixture was refluxed at 80 ℃ overnight and distilled under reduced pressure until completion. Dissolving the crude product in CH2Cl2Purification by silica gel column chromatography (PE/EtOAc ═ 1.25: 1) gave light yellow intermediate 1a (1.04g, 79.6% yield).
Intermediate 1a1H NMR(400MHz，DMSO-d6)δ10.23(s，1H)，9.87(s，1H)，8.47(s，1H)，7.90(s，1H)，7.36(dt，J＝7.0，2.1Hz，1H)，7.31-7.20(m，2H)，6.81(s，1H)，1.80(m，1H)，0.79(m，4H)。
Intermediate 1a (50mg, 0.17mmol) of a different substituted aniline (0.26mmol) was added to DMF. The reaction mixture was added dropwise to 3eq TFA and stirred at 60 ℃ overnight. Finally, saturated NaHCO is used3The mixture was quenched and extracted with EA and i-PrOH (10 mL. times.3). The combined organic layers were washed with anhydrous Na2SO4Drying, distilling under reduced pressure, and purifying by silica gel column chromatography (PE/EtOAc ═ 3: 1 to 1: 1) to obtain compounds 1 to 15.
1-15.
Compound 1, yield: 84.6%, M.p.235.9-236.7 ℃.1H NMR(400MHz，DMSO-d6)δ10.19(s，1H)，9.40(s，1H)，9.22(s，1H)，8.38-8.28(m，1H)，7.89(s，1H)，7.81(s，1H)，7.73(ddd，J＝8.3，2.4，1.2Hz，1H)，7.42(t，J＝7.9Hz，1H)，7.30-7.18(m，3H)，7.16-6.85(m，2H)，6.20(d，J＝1.1Hz，1H)，1.81(m，1H)，0.79(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.55，160.60，160.27，157.65，141.12，140.52，139.71，134.52，129.21，128.84，121.45，118.43，117.34，116.03，114.92，113.01，110.84，86.97，14.47，7.10(2C)。HRMS(ESI)+[C21H20F2N5O+]：calcd 396.1636。
Compound 2 yield: 88.4%. M.p.232.6-234.1 ℃.1H NMR(400MHz，DMSO-d6)δ10.12(s，1H)，9.04(s，1H)，8.45(s，1H)，8.18(s，1H)，7.79(t，J＝2.0Hz，1H)，7.26(dt，J＝7.8，1.8Hz，1H)，7.17(m，3H)，6.94(d，J＝2.7Hz，1H)，6.70(dd，J＝8.4，2.7Hz，1H)，5.89-5.82(m，1H)，3.32(s，3H)，2.12(s，3H)，1.80(m，1H)，0.92-0.66(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.50，161.43，160.74，157.70(2C)，140.84，139.58，138.82，138.30，131.14，128.68，124.58，114.47，112.65，111.08，110.44，84.91，55.00，17.00，14.44，7.06(2C)。HRMS(ESI)+[C22H24N5O2 +]：calcd 390.1930，found 390.1935.HPLC purity 97.288％(tR＝7.813min)。
Compound 3, yield: 74.6%. M.p.226.5-227.4 deg.C.1H NMR(400MHz，DMSO-d6)δ10.24(s，1H)，9.12(s，1H)，8.70(s，1H)，8.18(s，1H)，7.81(s，1H)，7.33-7.09(m，5H)，6.99(ddd，J＝9.4，6.9，2.5Hz，1H)，5.94(s，1H)，2.10(d，J＝2.1Hz，3H)，1.84(td，J＝7.4，3.7Hz，1H)，0.77(t，J＝5.5Hz，4H)。13C NMR(101MHz，DMSO-d6)δ171.64，162.19，161.30，160.71，159.79，157.67，140.66，139.62，128.67，126.64，121.23，119.73，114.65，112.81，111.20，110.58，85.42，14.41，9.79，7.07(2C)。HRMS(ESI)+[C21H21FN5O+]：calcd 378.1730，found 378.1722.HPLC purity 99.452％(tR＝7.935min)。
Compound 4, yield: 74.9%. M.p.249.6-250.8 ℃.1H NMR(400MHz，DMSO-d6)δ10.17(s，1H)，9.15(s，1H)，8.86(s，1H)，8.21(s，1H)，7.79(t，J＝2.0Hz，1H)，7.35-7.13(m，4H)，7.07(td，J＝8.3，1.8Hz，1H)，6.90(td，J＝8.1，1.5Hz，1H)，6.11(s，1H)，3.84(s，3H)，1.81(m，1H)，0.78(t，J＝5.5Hz，4H)。13C NMR(101MHz，DMSO-d6)δ171.78，160.80，160.67，157.58，147.79，147.70，145.98，143.55，140.53，139.55，128.81，128.02，123.61，116.76，114.88，110.83，86.19，56.05，14.47，7.13(2C)。HRMS(ESI)+[C21H20FN5O2Na+]：calcd 416.1499，found 416.1501。
Compound 5, yield: 92.9%. M.p.237.9-239.1 ℃.1H NMR(400MHz，DMSO-d6)δ10.18(s，1H)，9.15(d，J＝3.4Hz，2H)，8.26(s，1H)，7.80(t，J＝2.0Hz，1H)，7.42-7.32(m，1H)，7.30-7.15(m，3H)，7.14-7.06(m，2H)，6.17-6.14(m，1H)，3.82(s，3H)，1.82(m，1H)，0.78(dt，J＝8.7，2.8Hz，4H)。13C NMR(101MHz，DMSO-d6)δ171.56，160.55，160.46，157.65，148.03，146.86，145.66，140.62，139.68，137.20，128.80，115.43，114.80，112.92，110.73，106.07，86.21，55.75，14.45，7.09(2C)。HRMS(ESI)+[C21H20FN5O2Na+]：calcd 416.1499，found 416.1503。
Compound 6, yield: 81.3%. M.p.223.6-224.9 ℃.1H NMR(400MHz，DMSO-d6)δ10.21(s，1H)，9.32(s，1H)，9.21(s，1H)，8.30(s，1H)，7.81(d，J＝2.0Hz，1H)，7.43(d，J＝2.2Hz，1H)，7.34-7.14(m，5H)，6.22(s，1H)，3.83(s，3H)，1.87-1.78(m，1H)，0.78(dt，J＝7.8，2.9Hz，4H)。13C NMR(101MHz，DMSO-d6)δ171.57，160.55，160.21，157.60，154.37，140.91，140.53，139.70，129.50，128.80，114.87，112.98，112.95，112.07，110.78，104.05，86.96，55.75，14.45，7.10(2C)。HRMS(ESI)+[C21H21ClN5O2 +]：calcd 410.1384，found 410.1386。
Compound 7, yield: 81.3%, M.p.239.4-240.8 ℃.1H NMR(400MHz，DMSO-d6)δ10.25(s，1H)，9.18(d，J＝6.5Hz，2H)，8.27(s，1H)，7.81(s，1H)，7.26(m，3H)，7.21-7.12(m，2H)，7.11-7.05(m，1H)，6.52(dd，J＝8.3，2.5Hz，1H)，6.23(d，J＝1.1Hz，1H)，3.99(q，J＝6.9Hz，2H)，1.84(m，1H)，1.33(t，J＝6.9Hz，3H)，0.81-0.74(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.58，160.50，160.38，158.84，157.57，141.74，140.66，139.72，129.31，128.70，114.77，112.86，111.82，110.67，107.23，105.92，86.71，62.77，14.68，14.41，7.07(2C)。HRMS(ESI)+[C22H24N5O2 +]：calcd 390.1930，found 390.193。
Compound 8, yield: 79.5%, M.p.195.6-196.4 ℃.1H NMR(400MHz，DMSO-d6)δ10.17(s，1H)，9.38(s，1H)，9.20(s，1H)，8.31(s，1H)，8.12(t，J＝1.9Hz，1H)，7.93(dd，J＝8.2，2.3Hz，1H)，7.82-7.80(m，1H)，7.56(d，J＝7.6Hz，1H)，7.44(t，J＝7.9Hz，1H)，7.26-7.21(m，3H)，6.20(s，1H)，2.57(s，3H)，1.81(d，J＝5.3Hz，1H)，0.80-0.78(m，4H)。13C NMR(101MHz，DMSO-d6)δ197.84，171.57，160.59，160.31，157.82，141.00，140.54，139.69，137.33，129.03，128.85，123.92，121.52，118.53，114.90(2C)，110.81，86.84，26.69，14.47，7.11(2C)。HRMS(ESI)+[C22H22N5O2 +]：calcd 388.1773，found 388.1770。
Compound 9, yield: 81.3%. M.p.177.4-178.5 ℃.1H NMR(400MHz，DMSO-d6)δ10.16(s，1H)，9.09(m，2H)，8.26(s，1H)，7.80(t，J＝2.0Hz，1H)，7.41-7.32(m，2H)，7.21(m，4H)，6.83(d，J＝7.5Hz，1H)，6.18(s，1H)，2.58(q，J＝7.6Hz，2H)，1.81(m，1H)，1.18(t，J＝7.6Hz，3H)，0.79(dd，J＝6.7，4.2Hz，4H)。13C NMR(101MHz，DMSO-d6)δ171.56，160.57，160.53，157.66，144.28，140.66，140.32，139.67，128.81，128.59，121.43，119.30，117.40，114.84，112.88，110.73，86.08，28.28，15.62，14.47，7.09(2C)。HRMS(ESI)+[C22H24N5O+]：calcd 374.1981，found 374.1987。
Compound 10, yield: 77.4%. M.p.232.9-233.7 ℃.1H NMR(400MHz，DMSO-d6)δ10.17(s，1H)，9.16(d，J＝10.3Hz，2H)，8.28(s，1H)，7.81(s，1H)，7.30-7.15(m，5H)，7.14-7.09(m，1H)，6.55(dd，J＝8.1，2.5Hz，1H)，6.21(s，1H)，3.74(s，3H)，1.81(m，1H)，0.83-0.72(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.53，160.59，160.39，159.61，157.64，141.65，140.65，139.68，129.41，128.81，114.79，112.88，112.04，110.71，106.92，105.58，86.52，54.90，14.47，7.10(2C)。HRMS(ESI)+[C21H22N5O2 +]：calcd 376.1773，found 376.1779。
Compound 11, yield: 83.6%. M.p.163.9-165.7 ℃.1H NMR(400MHz，DMSO-d6)δ10.19(s，1H)，9.57(s，1H)，8.35(d，J＝0.9Hz，1H)，7.88(s，1H)，7.39-7.29(m，2H)，7.26-7.17(m，2H)，6.86(ddd，J＝8.3，2.5，0.9Hz，1H)，6.82-6.74(m，2H)，6.13(d，J＝0.9Hz，1H)，3.77(s，3H)，1.80(m，1H)，0.82-0.74(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.58，169.23，162.47，160.51，158.12，153.61，139.98，139.70，130.34，128.88，114.63，113.58，113.37，111.22，110.53，107.47，88.80，55.37，14.48，7.12(2C)。HRMS(ESI)+[C21H21N4O3 +]：calcd 377.1614，found 377.1618。
Compound 12, yield: 77.9%. M.p.195.6-196.4 ℃.1H NMR(400MHz，DMSO-d6)δ10.19(s，1H)，9.32(s，1H)，8.45-8.22(m，2H)，7.93(s，1H)，7.34(s，1H)，7.18(m，4H)，6.89(t，J＝7.4Hz，1H)，6.12(s，1H)，3.96(t，J＝8.6Hz，2H)，3.19(t，J＝8.6Hz，2H)，1.82(q，J＝6.3Hz，1H)，0.88-0.72(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.57，160.75，158.96，157.12，143.81，140.68，139.65，131.82，128.83，126.88，124.72，121.21，115.10，114.40，112.71，110.25，86.61，47.94，26.87，14.49，7.12(2C)。HRMS(ESI)+[C22H22N5O+]：calcd 372.1824，found 372.1821。
Compound 13, yield: 81.4%, M.p.201.3-202.5 ℃.1H NMR(400MHz，DMSO-d6)δ10.17(s，1H)，9.29(s，1H)，8.36(d，J＝0.9Hz，1H)，7.95-7.78(m，1H)，7.46(dd，J＝8.1，1.6Hz，1H)，7.33-7.22(m，1H)，7.17(dd，J＝5.0，1.7Hz，2H)，7.08-6.83(m，3H)，6.53(d，J＝1.0Hz，1H)，4.14(m，4H)，1.81(m，1H)，0.87-0.67(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.54，161.04，160.27，157.87，146.77，140.50，139.65，128.80，126.87，124.51，122.98，119.84，117.34，114.54，112.87，110.36，87.94，65.35，41.69，14.47，7.10(2C)。HRMS(ESI)+[C22H22N5O+]：calcd 388.1773，found 388.1768。
Compound 14, yield: 84.6 percent, M.p.209.7-210.5 ℃.1H NMR(400MHz，DMSO-d6)δ10.18(s，1H)，9.29(s，1H)，8.37(s，1H)，8.13(d，J＝8.1Hz，1H)，7.96-7.88(m，1H)，7.38-7.29(m，1H)，7.25-7.13(m，2H)，7.06(t，J＝7.8Hz，1H)，6.73(d，J＝7.5Hz，1H)，6.11(s，1H)，3.97(t，J＝8.5Hz，2H)，3.09(t，J＝8.5Hz，2H)，2.20(s，3H)，1.82(m，1H)，0.84-0.72(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.57，160.71，158.94，157.11，143.48，140.69，139.64，133.55，130.50，128.83，126.97，122.28，114.39，112.68(2C)，110.24，86.54，47.90，25.71，18.32，14.48，7.12(2C)。HRMS(ESI)+[C23H24N5O+]：calcd 386.1981，found 386.1990。
Compound 15, yield: 72.5%. M.p.231.4-232.5 ℃.1H NMR(400MHz，DMSO-d6)δ10.19(s，1H)，9.40(s，1H)，8.65(d，J＝8.2Hz，1H)，8.42(d，J＝0.8Hz，1H)，7.93(d，J＝1.3Hz，1H)，7.40-7.32(m，2H)，7.23-7.16(m，3H)，6.13(d，J＝1.0Hz，1H)，4.05(t，J＝8.7Hz，2H)，3.37(s，1H)，3.34-3.31(m，1H)，1.82(m，1H)，0.83-0.75(m，4H)。13C NMR(101MHz，DMSO-d6)δ171.58，160.91，158.84，157.11，145.43，140.48，139.66，129.83，128.86，128.12，125.08，122.91，118.68，117.15，114.48，112.88，110.37，87.19，48.06，25.87，14.48，7.12(2C)。HRMS(ESI)+[C23H21F3N5O+]：calcd 440.1698，found 440.1691。
Test example 1 compounds 1-15 were tested for their anti-proliferative activity on a549 cells.
Figure BSA0000272513520000081
aIC50Values were determined by cell viability for 48 hours.
Experimental results show that the compound provided by the invention has a good effect on inhibiting the proliferation activity of lung cancer cells, wherein the compound 1, 2, 4 and 10 has a good effect, and the optimized compound 10 has a good lung cancer cell inhibition activity.
Test example 2 compound 10 inhibited proliferation and colony formation studies in a549 cells.
A549 cells in the logarithmic growth phase are cultured by using culture media containing different concentrations of the compound 10, each group of cells are stained by crystal violet staining solution, and the cell growth and colony formation are observed. After 6 days of continuous culture, the results of colony formation experiments showed that Compound 10 significantly inhibited the proliferation and colony formation of A549 cells in a concentration-dependent manner at concentrations of 1nM, 5nM, 10nM, 15nM, and 20nM (FIG. 1A), and the colony formation was almost completely inhibited at a concentration of 20nM (FIG. 1B).
The experimental result shows that the compound 10 inhibits the proliferation and colony formation of A549 cells.
Experimental example 3 compound 10 inhibits tubulin polymerization the crystal structure that was studied in vitro and bound to tubulin.
One nitrogen atom of 4, 6-pyrimidine in compound 10 forms a critical hydrogen bond with D249 of β -tubulin, and the other nitrogen atom forms a hydrogen bond network with N101 and T179 of α -tubulin mediated by water molecules (fig. 2A).
Immunofluorescence assays were performed on a549 cells by confocal microscopy. As shown in FIG. 2B, the DMSO group microtubule network was in a normal arrangement, elongated, while the paclitaxel-treated tubulin was in an ordered aggregation and clustered, while the colchicine and Compound 10-treated tubulin were in a disordered and chaotic morphology and the effect of Compound 10 was more pronounced than that of colchicine (FIG. 2B).
The experimental results show that the compound 10 and colchicine of the present invention can interact with tubulin.
Test example 4 study of compound 10 on promotion of apoptosis in a549 cells.
A549 cells were subjected to annexin V-FITC/PI (AV/PI) double staining test, and the proportion of apoptotic cells after treatment with different concentrations of compound 10 was significantly increased in dose-dependent manner, 11.28%, 18.00% and 30.58% compared to the proportion of apoptotic cells in the control group (1.21%), respectively (FIG. 3A, FIG. 3B).
Western blot analysis showed that Bcl-2 was down-regulated after treatment with Compound 10, but Bad expression was up-regulated (FIG. 3C)
The experimental results show that the compound 10 of the present invention significantly induces apoptosis in a549 cells in a dose-dependent manner.
Test example 5 cell migration study of compound 10 in a549 cells.
After the A549 cells are treated by the compound 10 for 12h, the wound closure rate of the control cells is 29.5 +/-1.34%, and the recovery rates of the cells treated by the compound 10 are 23.2 +/-1.15%, 15.8 +/-0.87%, 9.8 +/-0.96% and 6.1 +/-0.50%, respectively (figure 4A and figure 4B).
The inhibitory effect of compound 10 on the migration ability of a549 cells was verified by a Transwell chamber assay. After treatment with compound 10 (5nM, 10nM, 20nM and 40nM), the migration number of A549 cells decreased with increasing drug concentration, migrating to 635 + -12, 486 + -10, 397 + -11 and 157 + -9 wound areas, which were significantly smaller than the control (724 + -16), respectively (FIG. 4C, FIG. 4D).
The experimental results show that the compound 10 of the invention obviously inhibits the in vitro tumor cell migration.

Claims (7)

1. The invention provides a compound shown as a formula I and a pharmaceutically acceptable salt thereof:
Figure FSA0000272513510000011
wherein R is1Is composed of
Figure FSA0000272513510000012
Figure FSA0000272513510000013
2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: the structural formula is shown as formula I.
3. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, wherein:
R1is composed of
Figure FSA0000272513510000014
Figure FSA0000272513510000015
Preferably R1Is composed of
Figure FSA0000272513510000016
4. The following are some preferred structures of the compounds of the present invention.
Figure FSA0000272513510000017
5. Use of a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of lung cancer.
6. Use according to claim 5, characterized in that: the lung cancer treatment medicine is an anti-tumor medicine; preferably, the anti-tumor drug is a tubulin inhibitor drug.
7. A pharmaceutical composition characterized by: a formulation comprising an effective amount of a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115850190A (en) * 2023-01-10 2023-03-28 四川大学 5-fluorouracil derivative and anti-tumor application thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115850190A (en) * 2023-01-10 2023-03-28 四川大学 5-fluorouracil derivative and anti-tumor application thereof

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