Isoindoline-piperidine carboxamide compound, preparation method and application thereof

Abstract

The present invention provides isoindolinyl-piperidinecarboxamide compounds, their preparation methods, and uses. The biologically active isoindolinyl-piperidinecarboxamide core groups contained in these compounds can be further chemically modified to produce a variety of compounds with higher biological activity, expanding the broad application of these compounds in biomedicine and the development of pharmaceutical preparations. These compounds significantly inhibit the proliferation of lung, liver, pancreatic, and gastric cancer cells at low (nanomolar) doses and effectively inhibit the growth of transplanted tumors in mice, demonstrating their potential for development as anti-tumor drugs.

Description

Technical Field

[0001] The present invention belongs to the field of tumor targeted therapy, and specifically relates to an isoindolinyl-piperidinecarboxamide compound, a preparation method and application thereof. Background Art

[0002] Cancer has a high incidence and mortality rate in my country, and the patients are becoming younger. Since the early symptoms of most cancers are not specific, most patients are already in the middle or late stages when diagnosed, with high rates of metastasis and recurrence after surgery. Traditional treatments such as combined chemotherapy and radiotherapy have significant side effects and low clinical benefit rates. Although great progress has been made in the diagnosis and treatment of some cancers in clinical practice, due to the inherent heterogeneity and phenotypic plasticity of cancer cells, many drugs have primary or secondary drug resistance, resulting in a lack of effective drugs available in the clinic. Therefore, the development of new cancer treatment drugs is of great significance.

[0003] This application synthesizes a novel class of isoindolinyl-piperidinecarboxamide compounds. Biological analysis has revealed that these compounds significantly inhibit the proliferation of lung, liver, pancreatic, and gastric cancer cells at extremely low doses, exhibiting excellent tumor-suppressing effects in animals without significant toxic side effects. Therefore, further development of these compounds will be of great significance for their application in tumor therapy. Summary of the Invention

[0004] The present invention aims to provide an isoindoline-piperidine carboxamide compound, a preparation method and application thereof.

[0005] Based on the above objectives, the present invention adopts the following technical solutions:

[0006] An isoindoline-piperidine carboxamide compound, the structural formula of which is shown in general formula I:

[0007]

[0008] Wherein, R1 is selected from H, F, Cl, Br, I, -CN, -CH3, -CF3, -OCH3, -OCF3, SO2NH2;

[0009] R2 is selected from H, F, Cl, Br, I, -CN, -CH3, -CF3, -OCH3, -OCF3, -COOH, OH,

[0010]

[0011] R3 is selected from H, F, Cl, Br, I, -CN, -CH3, -CF3, -OCH3, -OCF3, -COOH, OH; R4 is selected from H, F, Cl, Br, I, -CN, -CH3, -CF3, -OCH3, -OCF3, -COOH, OH; R5 is selected from H, F, Cl, Br, I, -CN, -CH3, -CF3, -OCH3, -OCF3, -COOH, OH; W, X, G are each independently selected from C or N;

[0012] K is selected from O or NH.

[0013] The above-mentioned isoindolinyl-piperidinecarboxamide compound is specifically a compound with the following structure:

[0014]

[0015]

[0016] A biologically acceptable salt formed by the above-mentioned isoindolinyl-piperidinecarboxamide compound and at least one of acetic acid, dihydrofolic acid, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanillic acid, tartaric acid, ascorbic acid, boric acid, lactic acid and ethylenediaminetetraacetic acid.

[0017] The preparation method of the above-mentioned isoindolinyl-piperidinecarboxamide compounds and the synthetic route are as follows:

[0018]

[0019] The specific synthesis steps are as follows:

[0020] (1) Compound 1, Compound 2, and potassium carbonate were dissolved in DMF, stirred at 65-75°C, and after the reaction was complete, the reaction solution was diluted with ethyl acetate and washed with saturated brine. The organic phase was dried by rotary evaporation to obtain Compound 3;

[0021] (2) Compound 3 was dissolved in methanol, and a solution of hydrogen chloride in ethyl acetate was added. After stirring at room temperature and the reaction was complete, the reaction solution was spin-dried to obtain compound 4;

[0022] (3) Compound 4, compound 6, HBTU and DIEA were dissolved in DMF and stirred at room temperature until the reaction was complete. The reaction solution was spin-dried and then subjected to column chromatography to obtain the target compound.

[0023] Furthermore, in step (1), the molar ratio of compound 1, compound 2 and potassium carbonate is 1:1:1.2; in step (2), the molar ratio of compound 3 and hydrogen chloride is 1:10; in step (3), the molar ratio of compound 4, compound 6, HBTU and DIEA is 1:1.2:1.2:3.

[0024] Application of the above-mentioned isoindolinyl-piperidinecarboxamide compounds and biologically acceptable salts thereof in the preparation of anti-tumor drugs.

[0025] Preferably, the anti-tumor drug refers to a drug for treating lung cancer, liver cancer, pancreatic cancer, gastric cancer, etc.

[0026] Specifically, the present invention synthesizes a class of isoindolinyl-piperidine carboxamide compounds IG230308B-1, IG230309B-1, IG230310B-1, IG230308C-1, IG230309C-1, IG230310C-1, IW230307A-1, IW230308A-1, IW230309A-1, IW230309B-1, IW230310B-1, IW230312B-1, IW230313A-1, IW23031 The inhibitory effects of these compounds on the proliferation of various cancer cells were determined using the CCK-8 assay. Their effects on the growth of liver cancer cells in vivo were also tested using a nude mouse xenograft model.

[0027] The results showed that the compounds of the present invention IG230308B-1, IG230309B-1, IG230310B-1, IG230308C-1, IG230309C-1, IG230310C-1, IW230307A-1, IW230308A-1, IW230309A-1, IW230309B-1, IW230310B-1, IW230312B-1, IW230313A-1, IW230313B-1, IW230313C-1, IG230320A-1, IG 230320B-1, IG230320C-1, IG230321A-1, IG230321B-1, IG230321C-1, IG230322A-1, IG230322B-1, IG230322C-1, IW230322A-1, IW230322B-1, IW230322C-1, IW230323A-1, IW230323B-1 and IW230323C-1 can effectively inhibit the proliferation of lung cancer, liver cancer, pancreatic cancer and gastric cancer cells, and significantly inhibit the growth of liver cancer in mice.

[0028] In summary, the present invention provides a new isoindolinyl-piperidinecarboxamide compound and its derivatives for use in tumor treatment and potential molecular mechanisms. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The blank solvent was used as a control to evaluate the anti-tumor effect of different doses of IG230308B-1 in mice. DETAILED DESCRIPTION

[0030] In order to make the technical purpose, technical solution and beneficial effects of the present invention more clear, the technical solution of the present invention is further described below in conjunction with the accompanying drawings and specific embodiments.

[0031] In the method for synthesizing the compound of formula I of the present invention, the various raw materials used in the reaction can be prepared by those skilled in the art based on prior knowledge, or can be prepared by methods known in the literature, or can be purchased commercially. The intermediates, raw materials, reagents, reaction conditions, etc. used in the above reaction schemes can be appropriately modified based on the prior knowledge of those skilled in the art.

[0032] In the present invention, unless otherwise specified: (i) the temperature is expressed in degrees Celsius (°C), and the operation is carried out at room temperature; more specifically, the room temperature refers to 20-30°C; (ii) the organic solvent is dried by a conventional drying method, and the solvent is evaporated by a rotary evaporator under reduced pressure, with the bath temperature not higher than 50°C; the developing solvent and the eluent are both in a volume ratio; (iii) the reaction process is monitored by thin layer chromatography (TLC); (iv) the final product has a satisfactory proton nuclear magnetic resonance ( 1 H-NMR).

[0033] Example 1: The synthesis of all compounds is as follows

[0034] The specific synthesis method takes compound IG230308B-1 as an example, and the structural formula is as follows:

[0035]

[0036] The compound IG230308B-1 is named (1-(4-(2,2,2-trifluoroethoxy)benzyl)piperidin-4-yl)(5-((5-(3-(4-(trifluoromethyl)phenyl)-1,2,4-oxadiazol-5-yl)pyrazin-2-yl)oxy)isoindolin-2-yl)methanone, and its synthesis route is as follows:

[0037]

[0038] Step 1.tert-butyl 5-((5-(3-(4-(trifluoromethyl)phenyl)-1,2,4-oxadiazol-5-yl)pyrazin-2-yl)oxy)isoindoline-2-carboxylate (Compound 3)

[0039] Compound 1 (2.0 g, 8.50 mmol, 1.0 eq), compound 2 (2.78 g, 8.50 mmol, 1.0 eq), and potassium carbonate (1.41 g, 10.2 mmol, 1.2 eq) were dissolved in 30 mL of DMF and reacted at 70°C for 3 hours. Completion of the reaction was monitored by TLC. The reaction solution was diluted with 200 mL of ethyl acetate and washed three times with saturated brine (200 mL x 3). The organic phase was dried and spin-dried. The crude product was purified by column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain 3.5 g of compound 3 as a white solid in a yield of 78.3%.

[0040] 1H NMR(CDCl3,300MHz)δ:9.05(d,J=1.1Hz,1H),8.70(d,J=1.1Hz,1H),8.38(d,J=8.2Hz,2H ),7.84(d,J=8.4Hz,2H),7.35(s,1H),6.95(d,J=10.0Hz,2H),4.83(s,4H),1.59(s,9H).

[0041] Step 2. (4-(2-(piperidin-1-yl)ethoxy)phenyl)methanol (Compound 4)

[0042] Compound 3 (3.2 g, 6.09 mmol, 1.0 eq) was dissolved in 20 mL of methanol and cooled to 0°C. A 4 M solution of hydrogen chloride in ethyl acetate (15.2 mL, 60.9 mmol, 10 eq) was added and the mixture was allowed to warm to room temperature for 2 hours. TLC monitoring indicated complete reaction with the formation of new spots. The reaction solution was then directly spin-dried to afford 2.75 g of compound 4 as a white solid, with a yield of 97.8%.

[0043] 1 H NMR (CDCl3, 300MHz) δ: 9.05 (d, J = 1.1 Hz, 1H), 8.70 (d, J = 1.1 Hz, 1H), 8.38 (d, J = 8. 2Hz,2H),7.84(d,J=8.4Hz,2H),7.35(s,1H),6.95(d,J=10.0Hz,2H),4.83(s,4H)

[0044] Step 3. (4-(4-(2,2,2-trifluoroethoxy)benzyl)piperazin-1-yl)(5-((5-(3-(4-(trifluoromethyl)phenyl)-1,2,4-oxadiazol-5-yl)pyrazin-2-yl)oxy)isoindolin-2-yl)methanone(IG230308B-1)

[0045] Compound 4 (1.0 g, 2.17 mmol, 1.0 eq), compound 6 (0.87 g, 2.60 mmol, 1.2 eq), HBTU (1.84 g, 2.60 mmol, 1.2 eq), and DIEA (0.84 g, 6.50 mmol, 3.0 eq) were dissolved in 25 mL of DMF and reacted at room temperature for 4 hours. Completion was monitored by TLC. The reaction solution was directly dried and passed through a column (DCM / MeOH = 60:1 to 20:1) to obtain 1.53 g of IG230308B-1 as a yellow solid in a 75% yield.

[0046] 1 H NMR(CDCl3,300MHz)δ:9.11(d,J=1.1Hz,1H),8.69(d,J=1.1Hz,1H),8.40(d,J=8.2Hz,2H),7.86(d,J=8.4Hz,2H),7.40(d,J=8.6Hz,2H),7 .37(s,1H),7.18(d,J=12.2Hz,2H),6.99(d,J=10.0Hz,2H),4.85(s,4H),4.41(s,3H),3.70(m,2H),3.61(s,2H),3.45(s,4H),2.55(s,4H).

[0047] Example 2, IG230308B-1, IG230309B-1, IG230310B-1, IG230308C-1, IG230309C-1, IG230310C-1, IW230307A-1, IW230308A-1, IW230309A-1, IW230309B-1, IW230310B-1, IW230312B-1, IW230313A-1, IW230313B-1, IW230313C-1, IG230320 Inhibitory effects of A-1, IG230320B-1, IG230320C-1, IG230321A-1, IG230321B-1, IG230321C-1, IG230322A-1, IG230322B-1, IG230322C-1, IW230322A-1, IW230322B-1, IW230322C-1, IW230323A-1, IW230323B-1 and IW230323C-1 on lung cancer, liver cancer, pancreatic cancer and gastric cancer cells

[0048] HGC827, HepG2, BxPC-3 and HGC-27 cells in the logarithmic growth phase were collected and counted, and the cell suspension concentration was adjusted to 5×10 4 / mL, added to a 96-well cell culture plate, with a volume of 100uL per well. Using DMSO as a solvent control, the compounds of the present invention, IG230308B-1, IG230309B-1, IG230310B-1, IG230308C-1, IG230309C-1, IG230310C-1, IW230307A-1, IW230308A-1, IW230309A-1, IW230309B-1, IW230310B-1, IW230312B-1, IW230313A-1, IW230313B-1, IW230313C-1, IG230320A-1, IG230320 B-1, IG230320C-1, IG230321A-1, IG230321B-1, IG230321C-1, IG230322A-1, IG230322B-1, IG230322C-1, IW230322A-1, IW230322B-1, IW230322C-1, IW230323A-1, IW230323B-1 and IW230323C-1 were diluted with DMSO and added to the culture wells so that the final concentrations of the compounds in the system were 0.1, 0.3, 1, 3, 10, 30, 100 and 300 (μmol / L), respectively. After 48 hours of culture, 10 μL of CCK-8 solution was added to each well and incubated at 37°C for 1 hour. The OD value at an absorption wavelength of 450 nm was measured using a microplate reader and the results were recorded. A cell growth curve was plotted with the compound dose as the abscissa and the absorbance as the ordinate. The statistical results of the half-maximal inhibition rate (IC50 value) of the compounds on tumor cells are shown in Table 1 below:

[0049] Table 1. CCK-8 detection of IG230308B-1, IG230309B-1, IG230310B-1, IG230308C-1, IG230309C-1, IG230310C-1, IW230307A-1, IW230308A-1, IW230309A-1, IW230309B-1, IW230310B-1, IW230312B-1, IW230313A-1, IW230313B-1, IW230313C-1, and IG230307A-1. Inhibitory effects of 20A-1, IG230320B-1, IG230320C-1, IG230321A-1, IG230321B-1, IG230321C-1, IG230322A-1, IG230322B-1, IG230322C-1, IW230322A-1, IW230322B-1, IW230322C-1, IW230323A-1, IW230323B-1, and IW230323C-1 on lung cancer, liver cancer, pancreatic cancer, and gastric cancer cells

[0050]

[0051]

[0052] The table shows: IG230308B-1, IG230309B-1, IG230310B-1, IG230308C-1, IG230309C-1, IG230310C-1, IW230307A-1, IW230308A-1, IW230309A-1, IW230309B-1, IW230310B-1, IW230312B-1, IW230313A-1, IW230313B-1, IW230313C-1, IG230320A- 1. IG230320B-1, IG230320C-1, IG230321A-1, IG230321B-1, IG230321C-1, IG230322A-1, IG230322B-1, IG230322C-1, IW230322A-1, IW230322B-1, IW230322C-1, IW230323A-1, IW230323B-1, and IW230323C-1 all exhibited favorable anti-proliferation effects against lung cancer, liver cancer, pancreatic cancer, and gastric cancer. This application further investigated the in vivo anti-tumor effects of this class of compounds using IG230308B-1 as an example.

[0053] Example 3: Antitumor efficacy of IG230308B-1 in mice

[0054] Thirty SPF-grade, 6-week-old BALB / c-nude female nude mice weighing 18-22 g were purchased from Spafford (Beijing) Biotechnology Co., Ltd. for tumor inoculation. The HepG2 cell transplanted tumor tissue that had been successfully inoculated subcutaneously was taken, the blood stains were washed with PBS, and it was cut into 1.5 mm small pieces. A small tumor piece was sucked out with a puncture needle, and after wiping the animal's abdomen with an alcohol cotton ball, it was directly pierced subcutaneously and injected into the tumor piece. The inoculated animals were returned to the original cage for breeding, and the environment was kept clean to avoid infection. One week after the tumor cells were inoculated, whether milky white nodules were formed at the inoculation site was observed. The long and wide diameters of the tumor were measured with a vernier caliper, and the volume of the tumor was calculated according to the formula (tumor volume = long diameter × short diameter 2 × 0.5). When the tumor volume reaches 100-150 mm 3 Grouping and dosing were performed at the same time. Twenty mice that met the experimental volume requirements were randomly divided into four groups of five mice each. IG230308B-1 was administered via oral gavage at doses of 5, 10, and 20 mg / kg per mouse. The suspension solvent (SUS Control, 2.5% CMC-Na aqueous solution) group served as the control. The mice were dosed once daily, 100 μL each time, for 11 consecutive days. The tumor volume and body weight of the mice were measured every other day. The results are detailed in the [Table 1]. Figure 1 .

[0055] Figure 1 The results of A showed that at the end of the experiment, the morphology of the transplanted tumor in the IG230308B-1 20 mg / kg group was significantly smaller than that in the solvent control group. Figure 1 The results of B showed that from the third day of administration, the transplanted tumor volume of the IG230308B-1 20 mg / kg group began to shrink and grew slowly in the later period, while the tumor volume of the control group and the other dosage groups continued to grow larger.

[0056] The above results show that IG230308B-1, IG230309B-1, IG230310B-1, IG230308C-1, IG230309C-1, IG230310C-1, IW230307A-1, IW230308A-1, IW230309A-1, IW230309B-1, IW230310B-1, IW230312B-1, IW230313A-1, IW230313B-1, IW230313C-1, IG230320A-1, IG230320B-1, I G230320C-1, IG230321A-1, IG230321B-1, IG230321C-1, IG230322A-1, IG230322B-1, IG230322C-1, IW230322A-1, IW230322B-1, IW230322C-1, IW230323A-1, IW230323B-1, and IW230323C-1 significantly inhibit the proliferation of lung, liver, pancreatic, and gastric cancer cells. IG230308B-1 significantly inhibits the growth of liver cancer xenografts in mice. Therefore, this class of drugs has promising anticancer effects and potential for development.

[0057] According to the general approach of drug development (conventional anti-tumor in vitro screening first, followed by targeted research), the compounds of the present invention can be applied to cancer treatment drugs related to abnormal cell proliferation, and can be prepared as anti-tumor drugs by mixing with human-acceptable salts or with pharmaceutical carriers.

[0058] Finally, it should be noted that the above embodiments are only used to illustrate and not to limit the technical solutions of the present invention. Any equivalent substitutions of the present invention and any modifications or partial substitutions that do not depart from the spirit and scope of the present invention should be included in the scope of protection of the claims of the present invention.

Claims

1. An isoindolinyl-piperidinecarboxamide compound, characterized in that: Specifically, the compound has the following structure:

2. A biologically acceptable salt of the isoindolinyl-piperidinecarboxamide compound of claim 1 and at least one of acetic acid, dihydrofolic acid, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanillic acid, tartaric acid, ascorbic acid, boric acid, lactic acid and ethylenediaminetetraacetic acid.

3. The method for preparing the isoindoline-piperidine carboxamide compound according to claim 1, wherein: The synthetic route is as follows: R1 is R2 is selected from F, Cl, I, R3 is selected from H, Cl; R4 is selected from H, F, Br; R5 is selected from H, Br; W, X, and G are independently selected from C or N; K is O; The specific synthesis steps are as follows: (1) Compound 1, Compound 2, and potassium carbonate were dissolved in DMF and stirred at 65-75°C. After the reaction was complete, the reaction solution was diluted with ethyl acetate and washed with saturated brine. The organic phase was dried by rotary evaporation and subjected to column chromatography to obtain Compound 3. (2) Compound 3 was dissolved in methanol, and a solution of hydrogen chloride in ethyl acetate was added. After stirring at room temperature and the reaction was complete, the reaction solution was spin-dried to obtain compound 4; (3) Compound 4, compound 6, HBTU and DIEA were dissolved in DMF and stirred at room temperature until the reaction was complete. The reaction solution was spin-dried and then subjected to column chromatography to obtain the target compound.

4. The method for preparing the isoindoline-piperidinamide compound according to claim 3, wherein: In step (1), the molar ratio of compound 1, compound 2 and potassium carbonate is 1:1:1.2; in step (2), the molar ratio of compound 3 and hydrogen chloride is 1:10; in step (3), the molar ratio of compound 4, compound 6, HBTU and DIEA is 1:1.2:1.2:

3.

5. Use of the isoindolinyl-piperidinecarboxamide compound and its biologically acceptable salt according to claim 1 or 2 in the preparation of antitumor drugs.

6. The use according to claim 5, characterized in that: The anti-tumor drug is a drug for treating lung cancer, liver cancer, pancreatic cancer and gastric cancer.

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