A selective ces2 inhibitor, methods of making the same, and uses thereof as a medicament

Abstract

The present application relates to a kind of containing effective amount of new type CES2 inhibitor shown in general formula (I), its preparation method and the pharmaceutical composition containing the compound, as the preparation treatment, prevention or mitigation one or more diseases or functional disorders, compared with prior art, it has more optimal in vivo pharmacological effect, has more broad application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to a novel selective CES2 inhibitor, a preparation method thereof and an application thereof, and belongs to the technical field of medicine. BACKGROUND

[0002] Human carboxylesterases (hCES) are ubiquitous phase I drug metabolizing enzymes that hydrolyze ester substances and activate their precursors. hCES mainly has two members: carboxylesterase 1 (hCES1) and carboxylesterase 2 (hCES2). hCES2 is mainly distributed in the intestine and liver, while hCES1 is mainly distributed in the liver. In addition, hCES1 and hCES2 have different physiological functions. hCES1 metabolizes exogenous substances, cholesterols from the liver and periphery, and proteins in the endoplasmic reticulum. hCES2 plays an extremely important role in the hydrolytic metabolism of various anti-tumor drugs with ester bonds or amide bonds such as capecitabine, irinotecan, flutamide, etc.

[0003] Irinotecan is a first-line treatment for some tumors. In the intestine, irinotecan is rapidly hydrolyzed by hCES2 into the toxic metabolite SN-38, which is considered to be the main source of side effects. This process leads to the accumulation of a large amount of SN-38, resulting in severe gastrointestinal (GI) toxicity. GI toxicity is manifested as early or delayed diarrhea, which occurs 24 hours after irinotecan administration. About 40%-80% of patients using irinotecan experience diarrhea, and the incidence of diarrhea is 100% at high doses. Among them, nearly 40% of patients experience severe and life-threatening diarrhea. The probability of developing late-onset diarrhea after treatment in patients with colorectal cancer treated with irinotecan is as high as 49%, and this symptom can last for 10 years. Therefore, severe late-onset diarrhea greatly limits the clinical application of irinotecan, and this side effect needs to be addressed urgently. However, although some patients experience relief from late-onset diarrhea after using LPA, LPA is not specific. LPA can cause severe constipation, abdominal pain, dizziness, nausea, and vomiting. High doses of LPA increase the incidence of intestinal obstruction. There are also reports that LPA has certain cardiotoxicity. In view of the deficiencies of LPA in efficacy and side effects, it is urgent to develop more effective and safer drugs for treating late-onset diarrhea.

[0004] The delayed diarrhea caused by irinotecan is closely related to the activity of hCES2, and it is reported that the combination of potent and selective hCES2 inhibitors can alleviate irinotecan-related toxicity. This strategy can block the hydrolysis of irinotecan in the human intestine, thereby reducing the exposure of intestinal SN-38, which can alleviate irinotecan-related diarrhea. Therefore, it is of great application prospect to find highly efficient and selective hCES2 inhibitors in alleviating prodrug-related toxicity or improving the bioavailability of hCES2-inactivated oral ester-containing drugs. In recent years, some clinical drugs or natural products have been reported to be identified as hCES2 inhibitors, including troglitazone, LPA, mulberroside C, glycyrrhetinic acid-30-ethyl ester, etc. However, these inhibitors (except LPA) are not applied to the clinic due to lack of effectiveness or acceptable safety.

[0005] Therefore, a novel, highly efficient and selective CES2 inhibitor is synthesized in this paper, which is superior to the commonly used clinical drug lomotil in intestinal disease treatment by verifying its intestinal disease treatment effect through in vitro inhibition of CES2 activity, irinotecan-induced delayed diarrhea and dextran sulfate sodium-induced ulcerative colitis model. SUMMARY

[0006] The purpose of the present application is to provide a CES2 inhibitor with good efficacy and high selectivity, and to provide a new potential drug for preventing or / and treating delayed diarrhea caused by chemotherapy drugs such as irinotecan, ulcerative colitis, chemotherapy diarrhea, Crohn's disease, traveler's diarrhea and other diseases. The present inventors have optimized the CES2 inhibitor described in the present application through a large number of research, practice and experience, which has unexpected excellent pharmacological characteristics. SUMMARY

[0008] In one aspect, the present application provides a compound having the following general formula (I):

[0009]

[0010] wherein:

[0011] A ring is selected from any of the following structures:

[0012] R is selected from chlorine or hydrogen.

[0013] The compound of the present application includes, but is not limited to:

[0014]

[0015] The compound of the present application is used for preparing a drug for treating a CES2-mediated disease.

[0016] Another aspect of the present application relates to a pharmaceutical composition comprising a therapeutically effective amount of the compound and a suitable carrier, diluent or excipient.

[0017] The present application also relates to the use of the compound for the preparation of a medicament for preventing / treating the side effects of chemotherapy drugs, irinotecan-induced early / late onset diarrhea, traveler's diarrhea, Crohn's disease, ulcerative colitis, large intestine inflammation, small intestine inflammation, colitis, ischemic colitis, acute / chronic diarrhea, functional diarrhea, irritable bowel syndrome, non-alcoholic fatty liver, alcoholic fatty liver, large intestine cancer and colon cancer, etc.

[0018] The compound of the present application can be synthesized by the following steps:

[0019]

[0020] As the base, inorganic bases and organic bases can be mentioned, for example, alkali metal carbonates such as sodium carbonate, potassium carbonate, cesium carbonate, etc.; alkali metal bicarbonates such as potassium bicarbonate, etc.; alkali metal hydroxides such as lithium hydroxide, sodium hydroxide, potassium hydroxide, etc.; and as the organic base, for example, triethylamine, pyridine, lutidine, n-butyllithium, potassium tert-butoxide, sodium methoxide, sodium ethoxide, etc. can be mentioned. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 I-12 (Compd. 12) reduces irinotecan-induced late onset diarrhea. (A) Changes in body weight of mice in each group during the administration period; (B) survival curve; (C) diarrhea score of mice in each group 24 h after the end of modeling; (D) fecal occult blood score of mice in each group 24 h after the end of modeling; (E) body weight loss score of mice in each group 24 h after the end of modeling compared with before modeling; (F) DAI score of mice in each group 24 h after the end of modeling; (G) percentage of body weight loss of mice in each group 24 h after the end of modeling compared with before modeling; (H) perianal condition of mice in each group. *p<0.05, **p<0.01, ***p<0.001 are the results of difference test of mice in the treatment group compared with mice in the NC group. # p<0.05, ## p<0.01, ### p<0.001 are the results of difference test of mice in the Model group compared with mice in the NC group.

[0022] Figure 2I-12 (Compd. 12) improved serum biochemistry of mice with irinotecan-induced delayed diarrhea. (A) ALP; (B) ALT; (C) AST; (D) TBIL; (E) BUN; (F) UA; (G) serum IL-1β; (H) serum IL-22. *p<0.05, **p<0.01, ***p<0.001 are the results of difference test of mice in treatment group compared with mice in NC group. # p<0.05, ## p<0.01, ### p<0.001 are the results of difference test of Model mice compared with mice in NC group.

[0023] Figure 3 I-12 (Compd. 12) relieved colonic inflammation of mice with delayed diarrhea. (A) HE staining results of colon tissue sections; (B) colon length; (C) histological score; (D) colon of mice. *p<0.05, **p<0.01, ***p<0.001 are the results of difference test of mice in treatment group compared with mice in Model group. # p<0.05, ## p<0.01, ### p<0.001 are the results of difference test of Model mice compared with mice in NC group.

[0024] Figure 4 I-12 (Compd. 12) relieved weight loss and diarrhea of mice with ulcerative colitis. (A) Body weight loss score; (B) diarrhea score; (C) fecal occult blood score; (D) DAI score; (E) percentage of body weight loss of mice in each group; (F) body weight change of mice during treatment. *p<0.05, **p<0.01, ***p<0.001 are the results of difference test of mice in treatment group compared with mice in Model group. # p<0.05, ## p<0.01, ### p<0.001 are the results of difference test of Model mice compared with mice in NC group.

[0025] Figure 5 I-12 (Compd. 12) reduced intestinal inflammation of mice with ulcerative colitis. (A) H&E staining and immunohistochemical staining of pathological sections of mice; (B) colon; (C) colon length; (D) Claudin-1 immunohistochemical staining score; (E) OCC immunohistochemical staining score; (F) H&E staining score; (G) histological score of colon. *p<0.05, **p<0.01, ***p<0.001 are the results of difference test of mice in treatment group compared with mice in Model group. # p<0.05,## p < 0.01, ### p < 0.001 indicates the difference between the Model mice and the NC group mice.

[0026] Figure 6 Effects of I-12 (Compd.12) on blood count and body weight in ICR mice. (A) ALT, (B) AST, (C) TBIL, (D) DBIL, (E) TG, (F) TC, (G) HDL-C, (H) LDL-C, (I) TBA, (J) BUN, (K) CR, (L) CK, (M) CK-MB, (N) GLU, (O) body weight.

[0027] Figure 7 Effects of I-12 (Compd.12) on organ indices in ICR mice. (A) Brain, (B) Heart, (C) Liver, (D) Spleen, (E) Lung, (F) Kidney as a percentage of mouse net weight (%). Detailed Implementation

[0028] The present invention will be further described below with reference to embodiments. It should be noted that the following embodiments are for illustrative purposes only and are not intended to limit the present invention. Various modifications made by those skilled in the art based on the teachings of the present invention should be within the scope of protection of the claims of this application.

[0029] Example 1

[0030] 8-Chloro-N-(4-Chlorophenyl)quinoline-2-amine (I-1)

[0031]

[0032] 1a (644.15 mg, 5.05 mmol), 2a (1 g, 5.05 mmol), Cs₂CO₃ (4.94 g, 15.15 mmol), palladium acetate (45.34 mg, 0.2 mmol), and Xantphos (116.87 mg, 0.2 mmol) were heated to 90 °C under a nitrogen atmosphere and reacted for three days. After the reaction was completed, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the solution was purified by column chromatography using a 20:1 column chromatography method to give 360 ​​mg of a relatively pure brown solid, with a yield of 24.66%.

[0033] 1H NMR (400 MHz, DMSO-d6) δ 9.85 (s, 1H), 8.22 (d, J = 8.9 Hz, 2H), 8.13 (d, J = 8.9 Hz, 1H), 7.79 - 7.71 (m, 2H), 7.39 (d, J = 8.9 Hz, 2H), 7.28 (t, J = 7.8 Hz, 1H), 7.14 (d, J = 8.9 Hz, 1H).13C NMR (101 MHz, DMSO-d6) δ 154.52, 143.28, 140.47, 137.97, 129.88, 128.91, 127.23, 125.29, 125.18, 123.41, 120.35, 115.36. TOF MS m / z: calcd. for C 15 H 10 Cl2N2 + [M+H] + : 288.0221; found 288.0226.

[0034] Example 2

[0035] 8-chloro-N-(p-tolyl)quinolin-2-amine (I-2)

[0036]

[0037] Following the procedure for the preparation of I-l, 180 mg of black brown solid was obtained in 26.53% yield.

[0038] 1 H NMR (400 MHz, Chloroform-d) δ 7.78 (d, J = 8.9 Hz, 1H), 7.63 - 7.60 (m, 2H), 7.49 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 7.9 Hz, 1H), 7.11 (dd, J = 8.0, 1.9 Hz, 3H), 6.85 (d, J = 8.9 Hz, 1H), 2.27 (s, 3H). 13 C NMR (101 MHz, DMSO) δ 154.80, 143.58, 139.02, 137.56, 130.60, 129.76, 129.74, 129.54, 127.17, 125.09, 122.98, 118.83, 115.36, 20.85. TOF MS m / z: calcd. for C 16 H 13 ClN2 + [M+H] + : 268.0767; found 268.0771.

[0039] Example 3

[0040] N-(4-chlorophenyl)quinolin-2-amine (I-3)

[0041]

[0042] Following the procedure for the preparation of I-1, a yellow solid was obtained. Yield 26.53%.

[0043] 1 H NMR (400 MHz, Chloroform-d) δ 7.83 (d, J = 8.8 Hz, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.56 (d, J = 8.0 Hz, 1H), 7.50 (d, J = 8.8 Hz, 3H), 7.22 (d, J = 8.7 Hz, 3H), 6.80 (d, J = 8.9 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 154.34, 147.18, 140.81, 137.54, 129.99, 128.88, 127.98, 126.82, 124.75, 124.07, 123.38, 120.22, 114.52. TOF MS m / z: calcd. for C15H11ClN2 + [M+H] + : 254.0611; found 254.0606.

[0044] Example 4

[0045] 8-chloro-N-(3-chlorophenyl)quinolin-2-amine (I-4)

[0046]

[0047] Following the procedure for the preparation of I-1, a yellow solid was obtained. Yield 26.53%.

[0048] 1 H NMR (400 MHz, DMSO-d6) δ 9.91 (s, 1H), 8.85-8.82 (m, 1H), 8.16 (d, J = 8.9 Hz, 1H), 7.82-7.71 (m, 3H), 7.38-7.28 (m, 2H), 7.15 (d, J = 8.9 Hz, 1H), 7.02 (dd, J = 7.8, 2.1 Hz, 1H). 13C NMR (101 MHz, DMSO-d6) δ 154.48, 143.27, 142.95, 138.05, 133.77, 130.48, 130.07, 129.93, 127.25, 125.37, 123.59, 121.18, 118.40, 117.10, 115.36. TOF MS m / z: calcd. for C 15 H 10 Cl2N2 + [M+H] + : 288.0221; found 288.0225.

[0049] Example 5

[0050] 8-chloro-N-(3-(trifluoromethyl)phenyl)quinolin-2-amine (I-5)

[0051]

[0052] Referring to the preparation method of I-1, 610 mg of black solid was obtained, with a yield of 49.91%.

[0053] 1 H NMR (400 MHz, Chloroform-d) δ 8.57 (s, 1H), 7.85 (d, J = 8.8 Hz, 1H), 7.78-7.73 (m, 1H), 7.66 (dd, J = 7.6, 1.3 Hz, 1H), 7.48 (dd, J = 8.0, 1.2 Hz, 1H), 7.37 (t, J = 7.9 Hz, 1H), 7.22 (d, J = 7.7 Hz, 1H), 7.20-7.13 (m, 1H), 6.84-6.76 (m, 2H). 13 C NMR (101 MHz, DMSO) δ 154.48, 143.16, 142.19, 138.19, 129.98, 129.96, 129.92, 127.28, 125.37, 123.73, 122.09, 117.84, 117.80, 115.31, 115.00, 114.96. TOF MS m / z: calcd. for C 16 H 10 ClF3N2 + [M+H] + : 322.0485; found 322.0489.

[0054] Example 6

[0055] 8-chloro-N-(M. tolyl)quinolin-2-amine (I-6)

[0056]

[0057] Following the procedure for the preparation of I-1, brownish black solid 390 mg was obtained in 57.48% yield.

[0058] 1 H NMR (400 MHz, Chloroform-d) δ 8.38 (d, J = 2.6 Hz, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.67 (dd, J = 7.6, 1.2 Hz, 1H), 7.53 - 7.48 (m, 3H), 7.31 (d, J = 8.8 Hz, 1H), 7.21 - 7.15 (m, 1H), 6.78 (d, J = 8.8 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 154.80, 143.50, 141.46, 138.16, 137.63, 129.86, 129.80, 128.92, 127.18, 125.14, 123.13, 122.46, 119.48, 115.90, 115.43, 21.89. TOF MS m / z: calcd. for C 16 H 13 ClN2 + [M+H] + : 268.0767; found 268.0774.

[0059] Example 7

[0060] 8-chloro-N-(3,4-dichlorophenyl)quinolin-2-amine (I-7)

[0061]

[0062] Following the procedure for the preparation of I-1, yellowish brown solid 330 mg was obtained in 40.93% yield.

[0063] 1 H NMR (400 MHz, Chloroform-d) δ 8.38 (d, J = 2.6 Hz, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.67 (dd, J = 7.6, 1.2 Hz, 1H), 7.53 - 7.48 (m, 3H), 7.31 (d, J = 8.8 Hz, 1H), 7.21 - 7.15 (m, 1H), 6.78 (d, J = 8.8 Hz, 1H). 13C NMR (101 MHz, DMSO-d6) δ 154.18, 143.05, 141.49, 138.23, 131.51, 130.65, 130.03, 130.00, 127.29, 125.40, 123.81, 122.66, 119.94, 118.77, 115.25. TOF MS m / z: calcd. for C 15 H9Cl3N2 + [M+H] + : 321.9831 ; found 321.9837.

[0064] Example 8

[0065] 8-Chloro-N-(3,5-dimethoxyphenyl)quinolin-2-amine (I-8)

[0066]

[0067] Following the procedure for the preparation of I-1, 380 mg of pink solid product was obtained in 46.51% yield.

[0068] 1 H NMR (400 MHz, DMSO-d6) δ 10.07 (s, 1H), 8.33 (s, 2H), 8.20 (d, J = 8.9 Hz, 1H), 7.84 - 7.75 (m, 2H), 7.33 (t, J = 7.8 Hz, 1H), 7.15 - 7.11 (m, 2H). 13 C NMR (101 MHz, DMSO) δ 154.14, 143.74, 143.03, 138.39, 134.43, 130.18, 130.05, 127.32, 125.53, 123.98, 120.48, 116.87, 115.27. TOF MS m / z: calcd. for C 15 H9Cl3N2 + [M+H] + : 321.9831 ; found 321.9837.

[0069] Example 9

[0070] 8-Chloro-N-(3-methoxyphenyl)quinolin-2-amine (I-9)

[0071]

[0072] Following the procedure for the preparation of I-1, 87 mg of brown solid was obtained in 7.65% yield.

[0073] 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.36 - 8.23 (m, 1H), 8.11 (d, J = 8.9 Hz, 1H), 7.76 (d, J = 7.3 Hz, 1H), 7.72 (d, J = 7.9 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.31 - 7.19 (m, 2H), 7.14 (d, J = 8.9 Hz, 1H), 6.55 (dd, J = 8.2, 2.5 Hz, 1H), 3.83 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 160.24, 154.84, 143.53, 142.79, 137.72, 129.84, 129.78, 129.67, 127.24, 125.26, 123.20, 115.59, 111.21, 107.95, 104.28, 55.52. TOF MS m / z: calcd. for C 16 H 13 ClN2O + [M+H] + : 4.0716; found 284.0720.

[0074] Example 10

[0075] 8-chloro-N-(4-(3-(methylsulfonyl)propoxy)phenyl)quinolin-2-amine (I-10)

[0076]

[0077] First Step

[0078] 1-(3-(methylsulfonyl)propoxy)-4-nitrobenzene (3b)

[0079] In a 250 ml round bottom flask, add 1b (0.5 g, 1.71 mmol), then add 2b (285.49 mg, 2.05 mmol) to the flask, finally add K2CO3 (709.07 mg, 5.13 mmol) as solvent in DMF, monitor the reaction of 1b complete by TLC plate, stop the reaction, quench the reaction with 6 times the amount of ice water, then extract, remove a large amount of 2b, column chromatography, purify 3b, the yield is close to 100%.

[0080] Second Step

[0081] 4-(3-(methylsulfonyl)propoxy)aniline (4b)

[0082] To 500 mg of 3b in a 250 ml round bottom flask, 50 mg of 10% Pb / C was added and reduced under hydrogen atmosphere to give compound 4b in near 100% yield using methanol as solvent.

[0083] Third step

[0084] 8-Chloro-N-(4-(3-(methylsulfonyl)propoxy)phenyl)quinolin-2-amine (I-10)

[0085] To 4b (400 mg, 1.74 mmol), 2a (345.49 mg, 1.74 mmol), Cs2CO3(1.71 g, 5.23 mmol), XantPhos (60.56 mg, 0.105 mmol), Pd(OCOCH3)2(23.50 mg, 0.10467 mmol) in a 250 ml round bottom flask, t-butanol was used as solvent, under nitrogen atmosphere, 90 °C, reaction for three days. Filtered, rotary evaporated, column chromatography to give yellow white solid I-10 100 mg, 14.67% yield.

[0086] 1 H NMR (300 MHz, DMSO-d6) δ 9.58 (s, 1H), 8.14 - 8.03 (m, 3H), 7.78 - 7.65 (m, 2H), 7.23 (t, J = 7.7 Hz, 1H), 7.08 (d, J = 9.0 Hz, 1H), 6.96 (d, J = 8.6 Hz, 2H), 4.08 (t, J = 6.2 Hz, 2H), 3.29 (t, J = 7.9 Hz, 2H), 3.03 (s, 3H), 2.14 (t, J = 7.7 Hz., 2H). 13 C NMR (75 MHz, DMSO-d6) δ 154.74, 153.38, 143.66, 137.49, 135.21, 129.74, 129.64, 127.19, 125.06, 122.83, 120.08, 115.34, 115.17, 66.29, 51.08, 40.67, 22.55. TOF MS m / z: calcd. for C 19 H 19 ClN2O3S+[M+H] + : 390.0805; found 390.0810.

[0087] Example 11

[0088] 8-Chloro-N-(2,3-dihydrobenzofuran-5-yl)quinolin-2-amine (I-11)

[0089]

[0090] Following the procedure for the preparation of 1-1, 300 mg of solid III was obtained in 62.5% yield.

[0091] 1 H NMR (300 MHz, DMSO-d6) δ 9.52 (s, 1H), 8.21 (d, J = 2.3 Hz, 1H), 8.04 (d, J = 8.9 Hz, 1H), 7.78 - 7.65 (m, 3H), 7.21 (t, J = 7.7 Hz, 1H), 7.06 (d, J = 8.9 Hz, 1H), 6.75 (d, J = 8.5 Hz, 1H), 4.50 (t, J = 8.6 Hz, 2H), 3.20 (t, J = 8.6 Hz, 2H). 13 C NMR (75 MHz, DMSO-d6) δ 154.81, 154.78, 143.69, 137.35, 134.84, 129.70, 129.55, 127.62, 127.13, 124.98, 122.66, 118.41, 116.47, 115.31, 108.94, 71.15, 30.11. TOF MS m / z: calcd. for C 17 H 13 ClN2O + [M+H] + : 296.0716; found 296.0722.

[0092] Example 12

[0093] N-(lH-Benzo[D]imidazol-2-yl)-8-chloroquinolin-2-amine (I-12)

[0094]

[0095] Following the procedure for the preparation of 1-1, 106 mg of yellow red solid was obtained in 23.82% yield.

[0096] 1 H NMR (300 MHz, DMSO-d6) δ 9.52 (s, 1H), 8.21 (d, J = 2.3 Hz, 1H), 8.04 (d, J = 8.9 Hz, 1H), 7.78 - 7.65 (m, 3H), 7.21 (t, J = 7.7 Hz, 1H), 7.06 (d, J = 8.9 Hz, 1H), 6.75 (d, J = 8.5 Hz, 1H), 4.50 (t, J = 8.6 Hz, 2H), 3.20 (t, J = 8.6 Hz, 2H). 13CNMR (75 MHz, DMSO-d6) δ 154.46, 150.41, 143.66, 142.35, 141.45, 132.02, 131.20, 131.13, 127.91, 127.73, 127.23, 123.38, 119.96, 116.86, 116.24, 110.90. TOF MS m / z: calcd for C 16 H 11 ClN4 + [M+H] + : 294.0672; found 294.0677.

[0097] Example 13

[0098] 8-Chloro-N-(lH-indol-5-yl)quinolin-2-amine (I-13)

[0099]

[0100] First Step

[0101] 1-Methyl-5-nitro-lH-indole (2c)

[0102] Take 0.5 mg 1c (1 eq), 870.44 mg KOH (5 eq) in a 250 ml round bottom flask, placed in an ice bath, add acetonitrile (5 ml / mm olI) as solvent, add 880.85 mg (2 eq) CH3I in the above reaction system, get 510 mg 2c, yield 93.30%.

[0103] Second Step

[0104] 1-Methyl-lH-indol-5-amine (3c)

[0105] Take 500 mg 2c in a 250 ml round bottom flask, add 50 mg 10% Pb / C in the above reaction system, add methanol as solvent, reduce under hydrogen atmosphere to get compound 3c, yield close to 100%.

[0106] Third Step

[0107] 8-Chloro-N-(lH-indol-5-yl)quinolin-2-amine (I-13)

[0108] To 3c 238.51 mg (1 eq), 2,8-dichloroquinoline 380 mg (1 eq), XantPhos 66.61 mg (0.06 eq), Cs2CO3 1.88 mg (3 eq), Pd(OCOCH3)2 225.85 mg (0.06), under nitrogen atmosphere, heat 90 °C, reaction three days. Get yellow solid 300 mg, yield 27%.

[0109] 1 H NMR (300 MHz, DMSO-d6) δ 9.58 (s, 1H), 8.75 (s, 1H), 8.07 (d, J = 8.9 Hz, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.74 - 7.67 (m, 2H), 7.44 (d, J = 8.7 Hz, 1H), 7.31 (d, J = 3.0 Hz, 1H), 7.25 (t, J = 7.7 Hz, 1H), 7.15 (d, J = 9.0 Hz, 1H), 6.42 (d, J = 3.0 Hz, 1H), 3.81 (s, 3H). 13 C NMR (75 MHz, DMSO-d6) δ 155.16, 143.90, 137.18, 133.97, 132.94, 130.26, 129.69, 129.60, 128.63, 127.13, 124.97, 122.52, 115.45, 114.96, 110.29, 109.97, 100.73, 32.98. TOF MS m / z: calcd. for C 18 H 14 ClN3 + [M+H] + : 307.0876; found 307.0882.

[0110] Example 14

[0111] The CES2 inhibitory activity, the in vivo alleviating irinotecan-induced delayed diarrhea activity, and the anti- ulcerative colitis activity of the compound of the present application can be determined by using the assay systems described below.

[0112] The following biological test examples describe and explain the present application.

[0113] The experimental methods of the specific conditions in the test examples of the present application are generally carried out under conventional conditions or according to the conditions recommended by the manufacturers of the products. The reagents for which the specific sources are not mentioned are the commonly used reagents purchased on the market.

[0114] Test Example 1 Inhibitory activity of the compound of the present application on CES1 and CES2

[0115] The present application uses the following method to determine the inhibitory activity of the compound of the present application on CES2:

[0116] In a reaction system with a total volume of 0.2 ml, HLM (2 μg / mL final concentration), PBS (0.1 M) buffer and the compound are added, and incubated at 37°C for 10 min, then the fluorescent probe substrate FD (15 μM) is added and incubated at 37°C for 20 min, then an equal volume of ice acetonitrile is added, and the reaction is terminated after vigorous shaking, and the protein is removed by centrifugation at 20000 x g, 4°C for 20 min. 200 μl of supernatant is taken to a 96-well plate. The absorbance of each well is detected by a fluorescence plate reader (Ex 480 nm, Em 520 nm). LPA is used as a positive control, and DMSO is used as a negative control. The DMSO in all solutions does not exceed 2%. The results are shown in Table 1.

[0117] Table 1: Inhibitory activity of CES2:

[0118]

[0119] The present application uses the following method to determine the inhibitory activity of the compound of the present application on CES1:

[0120] In a reaction system with a total volume of 0.1 ml, HLM (10 μg / mL final concentration), PBS (0.1 M, pH = 6.5) buffer and the inhibitor are added, and incubated at 37°C for 10 min, then the fluorescent probe substrate DME (3 μM) is added and incubated at 37°C for 10 min, then 50 μL of the reaction solution is added to an equal volume of LDR to terminate the reaction, and the absorbance of each well is detected at 580 nm. Bis (4-nitrophenyl) phosphate (BNPP) is used as a positive control. DMSO is used as a negative control, and the DMSO in all solutions does not exceed 2%. The results are shown in Table 2.

[0121] Table 2: Inhibitory activity of CES1:

[0122]

[0123] Conclusion: The compound I-12 of the present application has good inhibitory effect on CES2, and the in vitro activity is comparable to that of LPA. At the same time, the compound of the present application has high selectivity and does not inhibit the activity of CES1 in vivo.

[0124] The therapeutic effect of the compound of the present application on irinotecan-induced delayed diarrhea mice can be determined by using the following assay system:

[0125] 8-week-old SPF male C57BL / 6 mice (body weight 18-22 g) were randomly divided into 4 groups: 6 in the blank group, and 10 in each of the other groups. All animals were treated by gavage for ten days. On days 4-7, one hour after gavage, the blank group was injected intraperitoneally with 0.9% normal saline 5 ml / kg, and the other groups were injected intraperitoneally with irinotecan 75 mg / kg for 4 consecutive days. During the administration period, the body weight of the mice and the disease activity index (DAI, see Table 2) were recorded, and the results are shown in the accompanying Figure 1 ; after the end of the experiment, the mice were anesthetized, and blood was taken from the eyeball for detection of inflammatory indicators in the blood, and the results are shown in the accompanying Figure 2 ; colon tissue was taken for H&E staining to analyze the inflammation of the colon, and the results are shown in the accompanying Figure 3 .

[0126] The experimental results show that Compd. 12 can relieve irinotecan-induced delayed diarrhea, and this effect is much stronger than that of LPA (see accompanying Figure 1 ); I-12 administration can improve the serum inflammation of the mice, and this result is better than that of LPA (see accompanying Figure 2 ); the histological results show that Compd. 12 can improve the intestinal damage caused by irinotecan (see accompanying Figure 3 ).

[0127] In summary, I-12 can improve the severe delayed diarrhea of mice caused by the chemotherapy drug irinotecan, while LPA has almost no therapeutic effect in this disease model, which shows that I-12 has the potential to become the preferred therapeutic drug for delayed diarrhea.

[0128] Table 2: Disease activity index (DAI) score table:

[0129]

[0130] The therapeutic effect of the compound in the application on ulcerative colitis can be determined by using the determination system described as follows:

[0131] The animals were randomly divided into 6 groups: NC, Model, Mesalazine (20 mg / kg), I-12 (20 mg / kg), I-12 (40 mg / kg), and I-12 (60 mg / kg). Except for the NC group, the other groups were given 5% DSS drinking water daily for 7 days, during which gavage was given with the vehicle or drug treatment, and the DAI was recorded, and the results are shown in the accompanying Figure 4 ; at the end of the treatment, all animals were anesthetized, and blood was taken from the eyeball after 8 h of fasting, and were euthanized by decapitation. The colon was taken for pathological sectioning for H&E staining and immunohistochemical staining, and the results are shown in the accompanying Figure 5 .

[0132] The experimental results show that I-12 can alleviate the severity of ulcerative colitis, and this effect is much stronger than Mesalazine (see Figure 4 ) ; the colon length and the H&E staining and immunohistochemical staining results of the colon tissue show that the intestinal inflammation of the I-12 mice is less severe, and this result is comparable to that of Mesalazine (see Figure 5 ). In summary, I-12 has a good therapeutic effect on ulcerative colitis and is expected to become a treatment plan for different intestinal diseases in the clinic.

[0133] The animal subacute toxicity effect of the compound in the application can be determined by using the determination system described as follows:

[0134] ICR male mice were randomly divided into 6 groups (n = 6), namely a normal group and I-12 groups, and were respectively given blank solvent (0.5% CMC-Na) and I-12 (500 mg / kg) by gavage, the gavage volume was 0.1 mL / 10 g, once a day, for a total of 30 days. During the test, the mice were weighed at regular time and fixed point, and the general behavior changes, toxic conditions and death conditions of the ICR mice were observed. After the last administration on the 30th day and fasting for 8 hours, the mice were anesthetized and blood was collected, and the blood routine of the mice was detected, the results are shown in Table 1 Figure 6 ; then the brain, heart, liver, spleen, lung and kidney of the ICR mice were collected by dissection and weighed, and the results are shown in Table 2 Figure 7 .

[0135] The subchronic toxicity experiment results show that no animal died during the whole experiment, and there is no obvious difference between the body weight, diet and water, general performance and behavior, blood and blood biochemical results of the compound administration group and the normal group; the results of gross anatomy macroscopic observation and histological examination show that no obvious lesions are found in the organs of each dose group; I-12 is safe and non-toxic in the mouse body, and I-12 has no effect on the blood routine and body weight of the mice (see Table 3 Figure 6 ) ; and no obvious change is found in the organ index of the mice (see Table 4 Figure 7 ).

[0136] In summary, I-12 has great development prospects and is expected to be applied to the treatment of various intestinal diseases in the clinic.

Claims

1. A compound of general formula (I): ###0001### wherein: R1 is H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl, heterocyclyl, heterocyclylalkyl, haloalkyl, haloalkenyl, haloalkynyl, haloalkylalkyl, haloalkenylalkyl, haloalkynylalkyl, haloaryl, haloaralkyl, haloheteroaryl, haloheteroaralkyl, haloheterocyclyl, haloheterocyclylalkyl, -ORa, -SRa, -N(Ra)2, -CN, -C(O)Rb, -C(O)ORb, -C(O)N(Rb)2, -NRbC(O)Rb, -NRbC(O)ORb, -OC(O)Rb, -OC(O)N(Rb)2, -S(O)pRb, -S(O)pORb, -S(O)pN(Rb)2, -NRbS(O)pRb, -NRbS(O)pORb, -NRbS(O)pN(Rb)2, -P(O)(ORb)2, -P(O)(ORb)(Rb), -P(O)(Rb)2, -Si(Rb)3, - ​ A ring is selected from any one of the following structures: ​ ​

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