A tetrahydroquinoline compound containing an n-(hetero)aryl group and use thereof

Abstract

A composition comprising N The application belongs to the technical field of pharmacy and chemical industry. The (hetero)aryl-substituted tetrahydroquinolines are prepared by the method N The (hetero)aryl-substituted tetrahydroquinolines are evaluated for their inhibitory effect on HIF-1 signaling pathway and in-vitro anti-proliferative activity. The results show that the five modified compounds exhibit excellent HIF-1 inhibitory activity and have low toxicity. Unexpectedly, the compound I-11 shows strong anti-proliferative activity with a half-inhibitory concentration of 5.4 μM. Surface plasmon resonance experiments show that the K D of the compound I-4 with human Hsp90α protein is 4.7 μM, proving that the action target of I-4 is Hsp90. The compound and its pharmaceutically acceptable salts have a broad application space in the preparation of anti-tumor drugs and HIF-1 inhibitors.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmacy and chemical technology, more particularly, relates to a kind of N-(hetero) aryl substituted tetrahydroquinoline compound and application. BACKGROUND

[0002] Tetrahydroquinoline compounds exist widely in existing drugs and natural products, are an important class of nitrogen-containing heterocyclic skeleton, have antiviral, antibacterial, antimalarial and other wide range of biological activities. Research has found that compounds with tetrahydroquinoline as the skeleton exhibit excellent antitumor activity in DNA topoisomerase and microtubule protein and other multiple anticancer targets. Therefore, it is of great significance to study the antitumor activity of tetrahydroquinoline compounds.

[0003] Malignant tumor is still a serious problem that threatens physical health, and the incidence is increasing year by year. Although chemotherapy is a traditional method for treating cancer, the therapeutic effect of chemotherapy is getting worse due to its low selectivity, high toxicity and drug resistance. Therefore, it is urgent to develop efficient, low-toxicity and targeted antitumor drugs. The difference between cancer cells and normal cells lies in their unlimited proliferation ability. Because tumor cells need a large amount of oxygen and nutrients for rapid proliferation, anoxic environment is formed in the local area of tumor. Among the various signal pathways that cells adapt to hypoxic environment, HIF-1 plays a key role. HIF-1 is a heterodimeric protein formed by HIF-1α and HIF-1β, and HIF-1α is unique to HIF-1 and is regulated by intracellular oxygen concentration and determines the activity of HIF-1. The HIF-1 signal pathway activated by hypoxia can induce the expression of downstream genes including angiogenesis, glucose metabolism and invasion and migration, so inhibiting the activity of HIF-1 has become an effective and attractive strategy for treating various solid tumors. SUMMARY

[0004] In view of the problems existing in the prior art, the purpose of the present application is to provide a series of new N-(hetero) aryl substituted tetrahydroquinoline compounds with antitumor activity, a preparation method and application thereof in preparing antitumor drugs.

[0005] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0006] A kind of N-(hetero) aryl substituted tetrahydroquinoline compound, its structural formula is as shown in formula (I):

[0007]

[0008] R 1 is phenyl, phenethyl, benzyl containing substituent or not containing substituent, C1-C13 heteroaryl.

[0009] When the benzyl group has a substituent, the substituent on the benzyl group is a hydrogen, a C1-C3 alkyl group, a C1-C3 alkoxy group, a halogen, or the like. R 2 is a hydrogen, a halogen, a C1-C3 alkyl group, a C1-C3 alkoxy group, or the like; R 3 is a hydrogen, a C1-C3 alkyl group, a phenyl group, or the like; R 4 is a hydrogen, a C1-C3 alkyl group, a halogen, or the like.

[0010] In some specific compounds, R 1 is a benzyl group having a substituent, and the substituent is a fluorine, a methoxy group, or a methyl group;

[0011] R 2 , R 3 is a hydrogen;

[0012] R 4 is a hydrogen or a fluorine.

[0013] In some specific compounds, R 1 is a C1-C8 heteroaryl group.

[0014] In some specific compounds, R 1 is a thienyl group, a pyridyl group, a furanyl group, an indolyl group, or a pyrrolyl group, or the like.

[0015] A method for preparing a N-(hetero)aryl-substituted tetrahydroquinoline compound, comprising the following steps:

[0016]

[0017] wherein R 1 , R 2 , R 3 , R 4 are defined as in the structural formula (I), and R is a methyl group, a methoxy group, or a halogen.

[0018] (a) reacting compound 1 with a benzaldehyde (compound 2) having a different substituent to obtain an imine intermediate under the following conditions: using anhydrous magnesium sulfate as a water-removing agent, dichloromethane as a solvent, and under a nitrogen atmosphere, at room temperature, for 4-6 hours; wherein the molar ratio of compound 1 to compound 2 is 1.1:1, and the molar ratio of compound 1 to anhydrous magnesium sulfate is 1.1:2.

[0019] (b) reducing the imine intermediate to compound 3 under the following conditions: using sodium borohydride as a reducing agent, anhydrous methanol as a solvent, and under a nitrogen atmosphere, at 0-25°C; wherein the molar ratio of compound 1 to sodium borohydride is 1.1:2.2.

[0020] (c) under the conditions of room temperature, using anhydrous potassium carbonate as the acid-binding agent, N,N-dimethylformamide as the solvent, and under the atmosphere of nitrogen, compound 3 is reacted with 3-bromopropyne for 14-16 h to obtain compound 5; wherein the molar ratio of compound 3 to 3-bromopropyne is 1:1.1, and the molar ratio of compound 3 to anhydrous potassium carbonate is 1:2.

[0021] (d) under the conditions of room temperature, using anhydrous potassium carbonate as the acid-binding agent, acetonitrile as the solvent, and under the atmosphere of nitrogen, compound 6 is reacted with N-propargylaniline (compound 7) for 24-26 h to obtain compound 8; wherein the molar ratio of compound 7 to compound 6 is 1:1.5, and the molar ratio of compound 7 to anhydrous potassium carbonate is 1:2.

[0022] (e) under the conditions of room temperature, using anhydrous potassium carbonate as the acid-binding agent, N,N-dimethylformamide as the solvent, and under the atmosphere of nitrogen, compound 9 is reacted with compound 10 for 12-14 h to obtain compound 11; wherein the molar ratio of compound 9 to compound 10 is 1:1.5, and the molar ratio of compound 9 to anhydrous potassium carbonate is 1:2.

[0023] (f) under the conditions of 100-120℃, using cuprous bromide and triethylamine as the catalyst, 1,4-dioxane as the solvent, and under the atmosphere of nitrogen, compound 13 is reacted with formaldehyde and phenylacetylene for 12-14 h to obtain compound 14; wherein the molar ratio of compound 13 to formaldehyde, phenylacetylene is 1:1.5:3, the molar ratio of compound 13 to cuprous bromide is 1:0.2, and the molar ratio of compound 13 to triethylamine is 1:3.

[0024] (g) under the conditions of 110-120℃, using zinc acetate as the catalyst, dichloroethane as the solvent, and under the atmosphere of nitrogen, N-propargylaniline derivative (compound 5, 8, 11, 14) is reacted with compound 15 for 22-24 h to obtain compounds I-IV; wherein the molar ratio of N-propargylaniline derivative to compound 15 is 1:3, and the molar ratio of N-propargylaniline derivative to zinc acetate is 1:0.2-0.3.

[0025] The application relates to N-(hetero)aryl-substituted tetrahydroquinoline compounds for preparing antitumor drugs, which have low antiproliferative activity on tumor cells and can inhibit HIF-1 signal pathway activity by inhibiting Hsp90.

[0026] Compared with the prior art, the application has the beneficial effects that the application provides a N-(hetero)aryl substituted tetrahydroquinoline compound and application. On the basis of the characteristic structure, a series of N-(hetero)aryl substituted tetrahydroquinoline analogs are designed and synthesized. The MTT experiment results show that the compounds have certain anti-proliferation activity, and the double luciferase reporter gene detection shows that the compounds have significant HIF-1 signal pathway inhibition activity. When the methyl, fluorine atom and methoxy group are substituted at the para position of the benzyl benzene ring of the tetrahydroquinoline, the HIF-1 inhibition activity is obviously improved compared with the unsubstituted. When the benzyl benzene ring is modified at the ortho and meta positions, the HIF-1 inhibition activity is decreased, especially when the second methyl group is substituted, the HIF-1 inhibition activity is obviously decreased. When the benzyl benzene ring is replaced by a bioisostere thiophene ring, the HIF-1 inhibition activity is also excellent. When the benzyl group of the nitrogen atom is replaced by a phenethyl group, the HIF-1 inhibition activity is obviously decreased, which shows that the length of one carbon atom between the nitrogen atom and the benzene ring is the best. When the tetrahydroquinoline benzene ring is modified, it is found that when the methoxy group exists at the fifth position of the tetrahydroquinoline and the isopropyl group exists at the sixth position, the HIF-1 inhibition activity is slightly improved. It is found that when the indole is modified, when the methyl group exists at the fifth position of the indole, the HIF-1 inhibition activity is obviously improved, and when the bromine atom is substituted at the seventh position, the activity is obviously decreased. The surface plasmon resonance experiment proves that the direct target of the compound is Hsp90, and the CDOCKER docking analysis by using the DS software shows that the better HIF-1 inhibition activity compound and the target can produce hydrophobic or hydrogen bond interaction. It shows that the compounds have the potential as HIF-1 signal pathway inhibitors for treating cancer. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 is a binding affinity diagram of compound I-4 and hHsp90 alpha protein.

[0028] Figure 2 is a docking result diagram of compound I-1 and Hsp90 alpha inhibitor pharmacophore model 3R4N.

[0029] Figure 3 is a docking result diagram of compound I-2 and Hsp90 alpha inhibitor pharmacophore model 3R4N.

[0030] Figure 4 is a docking result diagram of compound I-3 and Hsp90 alpha inhibitor pharmacophore model 3R4N.

[0031] Figure 5 is a docking result diagram of compound I-4 and Hsp90 alpha inhibitor pharmacophore model 3R4N.

[0032] Figure 6 is a compound I-5 and Hsp90a inhibitor pharmacophore model 3R4N docking result figure.

[0033] Figure 7 is a compound I-8 and Hsp90a inhibitor pharmacophore model 3R4N docking result figure.

[0034] Figure 8 is a compound I-9 and Hsp90a inhibitor pharmacophore model 3R4N docking result figure.

[0035] Figure 9 is a compound I-12 and Hsp90a inhibitor pharmacophore model 3R4N docking result figure.

[0036] Figure 10 is a compound II-2 and Hsp90a inhibitor pharmacophore model 3R4N docking result figure. DETAILED DESCRIPTION

[0037] The following further describes the specific embodiments of the present application in combination with the technical solutions.

[0038] Example 1: Preparation of compounds I-1 to I-14

[0039] First step, preparation of compounds 3a-3h

[0040]

[0041] (a) To a solution of 1a (2.2 mmol) in anhydrous dichloromethane (4 ml) was added compound 2a (2 mmol) and anhydrous magnesium sulfate (481.4 mg) at room temperature. Then, the resulting mixture was stirred at room temperature for 4-8 hours, followed by vacuum filtration to obtain the filtrate, and concentrated in vacuum to obtain the imine crude product.

[0042] (b) A solution of the above imine crude product in anhydrous methanol (4 ml) was stirred at 0°C for 10 min at room temperature, followed by the addition of sodium borohydride 183.6 mg in batches. The resulting mixture was stirred at room temperature for 2-4 hours, and after the reaction was completed, saturated ammonium chloride solution was added to the reaction system until no gas bubbles were generated, followed by transfer to a separatory funnel with water and dichloromethane, and the organic phase was concentrated under reduced pressure to obtain the crude product 3f.

[0043] Referring to the above method, 3a-3c, 3g and 3h were synthesized by reacting 1a-1c with 2a-2d respectively, and the structural formulas of 1a-1c and 2a-2d are as follows:

[0044]

[0045] Second step, preparation of compounds 5a-5g

[0046]

[0047] (c) To a solution of 3a (2 mmol) in N,N-dimethylformamide (4 ml) was added propargyltribromide (3 mmol) and anhydrous potassium carbonate (552.8 mg) at room temperature. The reaction mixture was stirred at 80 °C for another 12-16 hours. After the completion of the reaction, the reaction mixture was transferred to a separatory funnel with ethyl acetate, the organic phase was extracted with water several times, and finally collected, washed with saturated brine and dried over anhydrous sodium sulfate, the organic phase was concentrated in vacuo, and the residue was purified by chromatography to give compound 5a as a yellow oil. Compounds 5b-5g were synthesized in turn by 3b-3g according to the procedure of step c (yield: 32-80%).

[0048] Third Step, Preparation of Compounds I-1-I-14

[0049] (g) To a solution of compound 5a (1 mmol) in anhydrous dichloroethane (1 ml) was added compound 15a (3 mmol) and anhydrous zinc acetate (36.8 mg) at room temperature. Then, the resulting mixture was stirred at 120 °C for 24-28 hours, after the completion of the reaction was detected by TLC, cooled at room temperature, transferred with DCM and filtered under reduced pressure, the organic phase was concentrated in vacuo, and the residue was purified by chromatography to give compound I-1 as a white solid, I-2-I-14 were synthesized by the reaction of 5a-5h with 15a-15g respectively according to the procedure of step g. The structures of 15a-15g are shown in the following formula:

[0050]

[0051] I-1: yield 69%, white solid. 1 H NMR (600 MHz, CDC13) δ 7.99 (s, 1H), 7.53 (d, J = 7.9 Hz, 1H), 7.38 (d, J = 8.2 Hz, 1H), 7.29 (t, J = 7.4 Hz, 2H), 7.25-7.17 (m, 4H), 7.09 (t, J = 7.5 Hz, 1H), 7.02 (dd, J = 12.0, 7.3 Hz, 2H), 6.97 (s, 1H), 6.62 (t, J = 7.2 Hz, 1H), 6.56 (d, J = 7.8 Hz, 1H), 5.04 (s, 1H), 4.43-4.70 (ABq, J = 17.4 Hz, 2H), 2.74 (dd, J = 19.3, 8.3 Hz, 1H), 2.66 (d, J = 15.8 Hz, 1H), 2.37-2.24 (m, 2H) ppm; HRMS (ESI Positive) m / z: calcd for C24 H 22 N2[M+H]+: 339.1783; found: 339.1854.

[0052] I-2: yield 17%, white solid. 1 H NMR (600 MHz, CDC13) δ 7.97 (s, 1H), 7.54 (d, J = 7.9 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.27-7.24 (m, 1H), 7.20 (t, J = 7.6 Hz, 1H), 7.14-7.04 (m, 2H), 7.02 (d, J = 1.3 Hz, 1H), 7.00 (d, J = 7.2 Hz, 1H), 6.97-6.90 (m, 2H), 5.02 (t, J = 3.8 Hz, 1H), 4.68-4.37 (ABq, J = 17.1 Hz, 2H), 2.83-2.53 (m, 2H), 2.37-2.26 (m, 1H), 2.25-2.15 (m, 1H): HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 345.1347; found: 345.14089.

[0053] I-3: yield 24%, white solid. 1 H NMR (600 MHz, CDC13) δ 7.92 (s, 1H), 7.52 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 8.2 Hz, 1H), 7.22-7.116 (m, 3H), 7.09 (dd, J = 9.9, 5.0 Hz, 1H), 7.05-6.98 (m, 2H), 6.98-6.92 (m, 2H), 6.89 (s, 1H), 6.65-6.59 (m, 1H), 6.51 (dd, J = 8.1, 2.9 Hz, 1H), 4.99 (s, 1H), 4.64-4.35 (ABq, J = 17.3 Hz, 2H), 2.79-2.57 (m, 2H), 2.38-2.13 (m, 2H): HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 357.1689; found: 357.1767.

[0054] I-4: yield 40%, white solid. 1H NMR (600 MHz, CDC13) δ 8.00 (s, 1 H), 7.52 (d, J = 7.9 Hz, 1 H), 7.38 (d, J = 8.1 Hz, 1 H), 7.21 (dd, J = 6.0, 2.2 Hz, 1 H), 7.15 (d, J = 8.7 Hz, 2 H), 7.12 - 7.06 (m, 1 H), 7.06 - 6.97 (m, 2 H), 6.94 (d, J = 1.9 Hz, 1 H), 6.86 - 6.79 (m, 2 H), 6.61 - 6.54 (m, 2 H), 5.02 (t, J = 4.0 Hz, 1 H), 4.65 - 4.34 (AB q, J = 17.1 Hz, 2 H), 3.78 (s, 3 H), 2.81 - 2.57 (m, 2 H), 2.36 - 2.19 (m, 2 H):

[0055] HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 369.1889; found: 369.19527.

[0056] I-5: Yield 44%, white solid. 1 H NMR (600 MHz, CDC13) δ 7.90 (s, 1 H), 7.53 (d, J = 6.1 Hz, 1 H), 7.35 (d, J = 6.8 Hz, 1 H), 7.27 (d, J = 5.6 Hz, 2 H), 7.20 (d, J = 8.5 Hz, 3 H), 7.09 (s, 1 H), 6.92 - 6.97 (m, 3 H), 6.48 (s, 1 H), 5.01 (s, 1 H), 4.66 - 4.39 (AB q, J = 17.1 Hz, 2 H), 2.78 - 2.63 (m, 3 H), 2.30 (s, 2 H), 1.21 (s, 6 H):

[0057] HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 381.2252; found: 381.23140.

[0058] I-6: Yield 26%, white solid. 1H NMR (600 MHz, CDC13) δ 7.99 (s, 1H), 7.52 (d, J = 7.9 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.29 (t, J = 7.5 Hz, 2H), 7.23 (d, J = 7.9 Hz, 2H), 7.22-7.18 (m, 2H), 7.09 (t, J = 7.6 Hz, 1H), 6.94 (d, J = 1.7 Hz, 1H), 6.91 (d, J = 8.1 Hz, 1H) 6.19 (dd, J = 8.1, 2.2 Hz, 1H), 6.14 (d, J = 2.0 Hz, 1H), 5.02 (t, J = 3.6 Hz, 1H), 4.67-4.41 (ABq, J = 17.4 Hz, 2H), 3.68 (s, 3H), 2.71-2.55 (m, 2H), 2.38-2.19 (m, 2H):

[0059] HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 369.1889; found: 369.19518.

[0060] I-7: Yield 38%, white solid (68.4 mg); 1H NMR (600 MHz, Chloroform-d) δ = 8.02 (s, 1H), 7.51 (d, J = 8.0, 1H), 7.38 (d, J = 8.3, 1H), 7.30 (dd, J = 8.7, 6.7, 3H), 7.21 (dd, J = 14.0, 7.2, 4H), 7.13-7.07 (m, 1H), 6.97 (s, 1H), 6.45 (s, 1H), 5.02 (d, J = 4.2, 1H), 4.62-4.43 (ABq, J = 17.2 Hz, 2H), 2.78-2.62 (m, 2H), 2.39-2.25 (m, 2H); HRMS (ESI) m / z Calcd for C24H21FN2 [M+H]+: 357.4444. Found: 357.1762.

[0061] I-8: Yield 33%, white solid. 1H NMR (600 MHz, CDC13) δ 8.06 (s, 1 H), 7.19 (d, J = 8.3 Hz, 2 H), 7.15 - 7.08 (m, 2 H), 7.04 - 6.98 (m, 2 H), 6.86 (d, J = 8.4 Hz, 2 H), 6.83 (s, 1 H), 6.80 - 6.73 (m, 1 H), 6.62 (t, J = 7.2 Hz, 1 H), 6.55 (d, J = 8.2 Hz, 1 H), 5.19 (s, 1 H), 4.67 - 4.42 (AB q, J = 17.1 Hz, 2 H), 3.80 (s, 3 H), 2.73 - 2.58 (m, 2 H), 2.38 (d, J = 11.3 Hz, 1 H), 2.30 (ddd, J = 17.5, 11.0, 6.4 Hz, 1 H).

[0062] MS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 387.17; found: 387.10.

[0063] I-9: Yield 55%, white solid. 1 H NMR (600 MHz, CDC13) δ 8.15 (s, 1 H), 7.29 (d, J = 7.9 Hz, 1 H), 7.16 (d, J = 8.5 Hz, 2 H), 7.06 (t, J = 7.7 Hz, 1 H), 7.04 - 6.98 (m, 2 H), 6.96 (s, 1 H), 6.94 - 6.89 (m, 1 H), 6.85 (d, J = 8.6 Hz, 2 H), 6.65 (t, J = 7.3 Hz, 1 H), 6.61 (d, J = 8.0 Hz, 1 H), 5.00 (s, 1 H), 4.68 - 4.34 (AB q, J = 17.1 Hz, 2 H), 3.80 (s, 3 H), 2.89 - 2.46 (m, 2 H), 2.43 - 2.11 (m, 2 H). MS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 387.17; found: 387.10.

[0064] I-10: Yield 44%, white solid. 1H NMR (600 MHz, CDC13) δ 7.91 (s, 1H), 7.45-7.33 (m, 1H), 7.13 (d, J = 7.8 Hz, 2H), 7.04-6.97 (m, 3H), 6.88-6.79 (m, 4H), 6.61 (t, J = 7.0 Hz, 1H), 6.57 (d, J = 8.0 Hz, 1H), 4.95 (s, 1H), 4.64-4.3 (ABq, J = 17.1 Hz, 2H), 3.77 (s, 3H), 7.98-7.84 (m, 2H), 2.25 (s, 2H).

[0065] MS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 387.17; found: 387.15.

[0066] I-11: Yellow solid, yield 27% (49.0 mg); 1 H NMR (600 MHz, Chloroform-d) δ = 8.10 (s, 1H), 7.39-7.27 (m, 4H), 7.23 (d, J = 7.7, 3H), 7.16-7.09 (m, 2H), 7.04 (dd, J = 11.3, 7.4, 2H), 6.94 (td, J = 9.0, 2.4, 1H), 6.70 (s, 2H), 4.96 (d, J = 4.6, 1H), 4.69-4.44 (ABq, J = 16.9 Hz, 2H), 2.73 (q, J = 7.5, 4.7, 2H), 2.31 (d, J = 9.6, 2H);

[0067] HRMS (ESI) m / z Calcd for C 24 H 21 FN2[M+H] + : 357.4444. Found: 357.1758.

[0068] I-12: Yellow solid, yield 28%. 1H NMR (600 MHz, Chloroform-d) δ 7.92 (s, 1H), 7.33-7.27 (m, 4H), 7.23 (dd, J = 8.0, 3.5 Hz, 3H), 7.02 (t, J = 8.9 Hz, 3H), 6.96 (s, 1H), 6.58 (s, 1H), 5.01 (d, J = 4.2 Hz, 1H), 4.69-4.44 (ABq, J = 17.3 Hz, 2H), 2.77-2.65 (m, 2H), 2.42 (s, 3H), 2.38-2.24 (m, 2H). HRMS (ESI) m / z Calcd for C 25 H 24 N2[M+H] + : 353.1939; found: 353.2003.

[0069] I-13: Yield 43%, white solid. 1 H NMR (600 MHz, CDCl3) δ 7.87 (s, 1H), 7.38 (dd, J = 6.8, 2.2 Hz, 1H), 7.32-7.26 (m, 2H), 7.23 (d, J = 7.2 Hz, 3H), 7.06-6.97 (m, 4H), 6.93 (d, J = 1.9 Hz, 1H), 6.61 (td, J = 7.3, 0.9 Hz, 1H), 6.54 (d, J = 8.1 Hz, 1H), 5.03 (dd, J = 4.3, 3.6 Hz, 1H), 4.70-4.41 (ABq, J = 17.5 Hz, 2H), 2.81-2.68 (m, 1H), 2.65 (dd, J = 11.8, 7.9 Hz, 1H), 2.48 (s, 3H), 2.29 (m, 2H): HRMS (ESI Positive) m / z: Calcd for C 24 H 22 N2[M+H] + : 353.1939; found: 353.2003.

[0070] Preparation of compounds II-1-II-2 of Example 2

[0071]

[0072] First step, preparation of compounds 8a-8c

[0073] (d) To a solution of N-propargylaniline (157.2 mg, 1.2 mmol) in acetonitrile (2.4 ml) was added commercially available benzyl bromide derivative and anhydrous potassium carbonate (331.7 mg) at room temperature. The resulting mixture was then stirred at room temperature for 24-28 hours, after TLC spotting indicated completion of the reaction, the reaction mixture was filtered under reduced pressure to obtain the filtrate, which was concentrated under reduced pressure and the residue was purified by chromatography to yield compound 8a-8c as a yellow oil.

[0074] Second step, preparation of compound II-1 - II-2

[0075] (g) To a solution of N-propargylaniline derivative (1 mmol) in anhydrous dichloroethane (1 ml) was added compound 15a (3 mmol) and anhydrous zinc acetate (36.8 mg) at room temperature. The resulting mixture was then stirred at 120 °C for 24-28 hours, after TLC spotting indicated completion of the reaction, it was cooled at room temperature, transferred to DCM and filtered under reduced pressure, the organic phase was concentrated under vacuum and the residue was purified by chromatography to yield compound as a white solid.

[0076] II-1 : yield 32%, white solid. 1 H NMR (400 MHz, CDC13) δ 7.95 (s, 1H), 7.53 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 8.1 Hz, 1H), 7.29 - 7.22 (m, 3H), 7.19 (t, J = 7.1 Hz, 3H), 7.16 - 7.12 (m, 2H), 7.09 (t, J = 7.5 Hz, 1H), 6.98 (d, J = 6.7 Hz, 1H), 6.85 (d, J = 2.2 Hz, 1H), 6.79 (d, J = 8.2 Hz, 1H), 6.62 (t, J = 7.3 Hz, 1H), 4.71 (t, J = 4.2 Hz, 1H), 3.75 - 3.65 (m, 1H), 3.43 - 3.31 (m, 1H), 3.01 - 2.77 (m, 2H), 2.66 - 2.55 (m, 2H), 2.22 (dq, J = 13.1, 4.5 Hz, 1H), 2.14 - 1.92 (m, 1H):

[0077] HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 353.1939; found: 353.2034.

[0078] II-2: yield 31%, white solid. 1H NMR (600 MHz, CDC13) δ 7.95 (s, 1 H), 7.52 (d, J = 7.8 Hz, 1 H), 7.36 (d, J = 8.2 Hz, 1 H), 7.19 (t, J = 7.6 Hz, 1 H), 7.16 - 7.06 (m, 5 H), 7.01 (dd, J = 17.1, 7.9 Hz, 2 H), 6.92 (d, J = 2.0 Hz, 1 H), 6.60 (t, J = 7.3 Hz, 1 H), 6.55 (d, J = 8.2 Hz, 1 H), 5.02 (s, 1 H), 4.67 - 4.37 (AB q, J = 17.4 Hz, 2 H), 2.80 - 2.68 (m, 1 H), 2.63 (dd, J = 15.8, 3.6 Hz, 1 H), 2.32 (s, 3 H), 2.31 - 2.22 (m, 2 H):

[0079] HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 353.1939; found: 353.2020.

[0080] Preparation of compounds III-1-III-2 of Example 3

[0081]

[0082] First step, preparation of compounds 11a-11b

[0083] (e) To a solution of aniline derivative substituted with nitrogen alkyl (2 mmol) in N,N-dimethylformamide (4 ml) was added bromo-propyne or butyne (3 mmol) and anhydrous potassium carbonate (552.5 mg) at room temperature. The reaction mixture was stirred at 80 °C for 12-16 hours. After the completion of the reaction, the reaction mixture was transferred to a separatory funnel with ethyl acetate, the organic phase was extracted with water several times, and finally collected, washed with saturated brine and dried over anhydrous sodium sulfate, and the organic phase was concentrated in vacuo, and the residue was purified by chromatography to give compound 11a-11b as a yellow oil.

[0084] Second step, preparation of compounds III-1-III-2

[0085] (g) To a solution of N-propargylaniline derivative (1 mmol) in dry dichloroethane (1 ml) was added compound 15a (3 mmol) and anhydrous zinc acetate (36.8 mg) at room temperature. The resulting mixture was then stirred at 120 °C for 24-28 h, cooled at room temperature after TLC monitoring of the complete reaction, transferred with DCM and filtered under reduced pressure, the organic phase was concentrated in vacuo and the residue was purified by chromatography to yield compound as a white solid.

[0086] III-1 : yield 16%, white solid. 1 H NMR (600 MHz, CDC13) δ 7.80 (s, 1H), 7.59 (d, J = 7.9 Hz, 1H), 7.31 (d, J = 8.1 Hz, 1H), 7.26 (d, J = 7.9 Hz, 2H), 7.24-7.16 (m, 3H), 7.11 (t, J = 7.5 Hz, 1H), 7.02-6.93 (m, 4H), 6.91 (s, 1H), 6.71 (d, J = 14.2 Hz, 1H), 5.27 (d, J = 3.3 Hz, 1H), 2.70 (d, J = 16.2 Hz, 1H), 2.65-2.54 (m, 1H), 2.43-2.24 (m, 2H):

[0087] MS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H]+: 325.16; found: 354.94

[0088] III-2: yield 43%, white solid. 1 H NMR (600 MHz, CDC13) δ 7.99 (s, 1H), 7.54 (d, J = 7.9 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.31-7.26 (m, 2H), 7.23-7.18 (m, 4H), 7.16 (d, J = 7.5 Hz, 1H), 7.11-7.07 (m, 1H), 7.05-7.01 (m, 1H), 6.96 (d, J = 2.4 Hz, 1H), 6.67 (t, J = 7.3, 0.9 Hz, 1H), 6.55 (d, J = 6.5 Hz, 1H), 5.02 (t, J = 4.2 Hz, 1H), 4.42-4.70 (ABq, J = 17.5 Hz, 2H), 2.84 (qt, J = 11.0, 5.5 Hz, 1H), 2.30 (ddd, J = 11.5, 7.8, 3.7 Hz, 1H), 2.06 (ddd, J = 12.8, 10.9, 4.9 Hz, 1H), 1.32 (d, J = 6.8 Hz, 3H):

[0089] HRMS (ESI Positive) m / z: calcd for C 24 H22N2[M+H]+: 353.1939; found: 353.2008

[0090] Preparation of compound IV of example 4

[0091]

[0092] First step, preparation of compound 14a

[0093] (f) To a solution of 3a (1.5 mmol) in 1.4-dioxane (3 ml) was added formaldehyde (2.2 mmol) and phenylacetylene (4.5 mmol), triethylamine (4.5 mmol) at room temperature. The reaction mixture was stirred at 110 °C for another 10-14 hours. After the completion of the reaction, the reaction mixture was transferred with ethyl acetate and filtered under reduced pressure, the organic phase was concentrated in vacuum and the residue was purified by chromatography to yield compound 14a as a yellow oil.

[0094] Second step, preparation of compound IV

[0095] (g) To a solution of compound 14a (1 mmol) in dry dichloroethane (1 ml) was added compound 15a (3 mmol) and anhydrous zinc acetate (36.8 mg) at room temperature. Then, the resulting mixture was stirred at 120 °C for 24-28 hours, after the completion of the reaction as monitored by TLC, it was cooled at room temperature, transferred with DCM and filtered under reduced pressure, the organic phase was concentrated in vacuum and the residue was purified by chromatography to yield compound IV as a white solid.

[0096] IV: yield 43%, white solid. 1 H NMR (600 MHz, CDC13) δ 7.97 (s, 1H), 7.49 (d, J = 7.9 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.30 (t, J = 7.4 Hz, 3H), 7.27 (dd, J = 9.7, 4.8 Hz, 4H), 7.24 - 7.17 (m, 4H), 7.14 (dd, J = 9.9, 8.6 Hz, 3H), 7.09 - 7.02 (m, 2H), 6.97 (d, J = 2.1 Hz, 1H), 6.63 (dt, J = 12.6, 5.9 Hz, 2H), 6.55 - 6.51 (m, 1H), 5.02 (t, J = 4.2 Hz, 1H), 4.72 (d, J = 17.2 Hz, 1H), 4.44 (d, J = 17.3 Hz, 1H), 3.99 (t, J = 7.6 Hz, 1H), 2.62 - 2.48 (m, 2H),:

[0097] HRMS (ESI Positive) m / z: calcd for C 24 H 22 N2[M+H] + : 415.2096; found: 415.2156.

[0098] The structural formula of the N-(hetero)aryl substituted tetrahydroquinoline analogues synthesized in the embodiments of the present application is as follows:

[0099]

[0100] Application Example 1

[0101] Anti-proliferation effect of the N-(hetero)aryl substituted tetrahydroquinoline analogues of the present application on tumor cells

[0102] Test method: The ultraclean table was treated with ultraviolet for 30 min in advance, and all the articles were used after being wiped with alcohol. The Hela cells in the logarithmic growth phase were counted after trypsin digestion, and the cell density was adjusted to 2×10 4 3 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, and 100 μl of each was added to the cell culture plate. After 48 h of normoxic culture, 20 μl of MTT (5 mg / ml) was added to each well under light shielding, and the incubation was continued for 4 h. Then the culture solution was removed, 200 μl of chemically pure DMSO was added to each well, and the absorbance at 570 nm was tested using an enzyme marker. Cell survival rate = [(relative OD value of the control group - relative OD value of the experimental group) / relative OD value of the control group] × 100%, and the IC 50 value of the drug was calculated using IBM SPSS Statistics software.

[0103] The anti-proliferation experimental results of the N-(hetero)aryl substituted tetrahydroquinoline analogues of the present application on tumor cells are shown in Table 1.

[0104] Table 1 Anti-proliferation activity of molecules

[0105]

[0106]

[0107] ​The MTT experiment of the synthesized N-(hetero)aryl substituted tetrahydroquinoline compounds showed that, compared with the previously published molecule I-1, most of the compounds had moderate level of in vitro anti-proliferation activity, and unexpectedly, the compound I-10 showed strong anti-proliferation activity, and the half-inhibitory concentration was 5.4±0.8 μM.

[0108] Application Example 2

[0109] Inhibition effect of N-(hetero)aryl substituted tetrahydroquinoline analogues on HIF-1 signal pathway

[0110] Test method: using lentivirus-mediated stable transfection dual-luciferase (firefly luciferase, Renilla luciferase) Hela cell line, after recovery and stabilization, logarithmic growth period Hela cells were digested and counted, and adjusted to a cell density of 2.5×10 5 4 / ml, 100 μL of cell suspension was added to a white opaque 96-well white plate, and cultured under normal oxygen condition for 12 h, after observing cell adhesion, 100 μL of culture solution containing 20 μM drug was added to each well, and continued to be cultured under normal oxygen condition for ih, and then transferred to low oxygen culture for 12 h. Then the normal oxygen plate and the low oxygen plate were taken out, first the culture solution was aspirated, washed with 1×PBS for 2-3 times, after aspirating and washing with PBS, 20 μL of PLB lysis solution was added to each well, and shaken for 20 min at room temperature in the dark. 50 μL of firefly and Renilla luciferase substrate was added to each well using the automatic sample adding system of the enzyme label instrument, and the chemiluminescence was detected by end-point method. The Renilla luciferase luminescence value was used as an internal reference, and the inhibition rate was calculated.

[0111] Transcriptional inhibition results of N-(hetero)aryl substituted tetrahydroquinoline analogues of the application on hypoxia-inducible factor HIF-1:

[0112] Table 2 HIF-1 inhibition activity of molecules

[0113]

[0114]

[0115] The synthesized N-(hetero)aryl substituted tetrahydroquinoline compounds were subjected to HIF-1 inhibition experiment under non-anti-proliferation concentration, and the results showed that, compared with the previously published molecule I-1, the compounds I-3, I-4, I-8, I-9 and II-2 showed better HIF-1 inhibition activity, in particular, the HIF-1 inhibition activity of I-4 was obviously improved, and the introduction of methoxy group was the key to the HIF-1 inhibition activity of the compound.

[0116] Application Example 3

[0117] The N-(hetero)aryl substituted tetrahydroquinoline analogues of the present application are tested for interaction with Hsp90a protein: Hsp90a recombinant protein (200 μg / mL, 10084-H08H) is coupled to a CM5 chip using amino coupling. The coupling process of the recombinant protein is performed according to the protein coupling calculation formula provided by GE Company on a Biacore T200 surface plasmon resonance instrument, and the final protein coupling amount is 10000 RU. Compound I-4 is tested for in vitro binding force with the HSP90a recombinant protein CM5 chip. The test compound is prepared into a stock solution with a concentration of 10 mM, and the final concentration is diluted to 0.1 μM, 0.2 μM, 0.39 μM, 0.78 μM, 1.56 μM, 3.125 μM, 6.25 μM using running buffer, 200 μL of each solution. In the experiment, 17AAG is used as a positive control, and 10 mM DMSO stock solution is diluted to a concentration of 0.31 μM, 0.625 μM, 1.25 μM, 2.5 μM, 5 μM. The positive control can bind to the CM5 chip coupled with Hsp90a recombinant protein, indicating the accuracy of the test results. The Kd value of the compound is calculated by surface plasmon resonance experiment, and the specific results are shown in Table 1. D Table 1 Figure 1 .

[0118] The N-(hetero)aryl substituted tetrahydroquinoline analogues of the present application are tested for interaction with Hsp90a protein: Hsp90a recombinant protein (200 μg / mL, 10084-H08H) is coupled to a CM5 chip using amino coupling. The coupling process of the recombinant protein is performed according to the protein coupling calculation formula provided by GE Company on a Biacore T200 surface plasmon resonance instrument, and the final protein coupling amount is 10000 RU. Compound I-4 is tested for in vitro binding force with the HSP90a recombinant protein CM5 chip. The test compound is prepared into a stock solution with a concentration of 10 mM, and the final concentration is diluted to 0.1 μM, 0.2 μM, 0.39 μM, 0.78 μM, 1.56 μM, 3.125 μM, 6.25 μM using running buffer, 200 μL of each solution. In the experiment, 17AAG is used as a positive control, and 10 mM DMSO stock solution is diluted to a concentration of 0.31 μM, 0.625 μM, 1.25 μM, 2.5 μM, 5 μM. The positive control can bind to the CM5 chip coupled with Hsp90a recombinant protein, indicating the accuracy of the test results. The Kd value of the compound is calculated by surface plasmon resonance experiment, and the specific results are shown in Table 1. D Table 1 D The N-(hetero)aryl substituted tetrahydroquinoline analogues of the present application are tested for interaction with Hsp90a protein: Hsp90a recombinant protein (200 μg / mL, 10084-H08H) is coupled to a CM5 chip using amino coupling. The coupling process of the recombinant protein is performed according to the protein coupling calculation formula provided by GE Company on a Biacore T200 surface plasmon resonance instrument, and the final protein coupling amount is 10000 RU. Compound I-4 is tested for in vitro binding force with the HSP90a recombinant protein CM5 chip. The test compound is prepared into a stock solution with a concentration of 10 mM, and the final concentration is diluted to 0.1 μM, 0.2 μM, 0.39 μM, 0.78 μM, 1.56 μM, 3.125 μM, 6.25 μM using running buffer, 200 μL of each solution. In the experiment, 17AAG is used as a positive control, and 10 mM DMSO stock solution is diluted to a concentration of 0.31 μM, 0.625 μM, 1.25 μM, 2.5 μM, 5 μM. The positive control can bind to the CM5 chip coupled with Hsp90a recombinant protein, indicating the accuracy of the test results. The Kd value of the compound is calculated by surface plasmon resonance experiment, and the specific results are shown in Table 1.

[0119] Application Example 4

[0120] N-(hetero)aryl substituted tetrahydroquinoline analogues and Hsp90a protein inhibitor pharmacophore model interaction mode analysis

[0121] Test method: DS software uses CDOCKER docking method.

[0122] Small molecule preparation: In "Tools Explore", expand Small Molecules\Prepare or Filter Ligands, click Prepare Ligands command, after completion, in "Tools Explore", expand Small Molecules\Minimize Ligands, click Full Minimization command, the "Minimize Ligands" dialog box is popped up. Click "Run" after setting is completed, start running.

[0123] Macromolecule acquisition: PDB database, select Hsp90α inhibitor pharmacophore model 3R4N

[0124] Macromolecule preparation: In "Tools Explore", expand Macromolecules\Prepare Protein, click Prepare Protein command, the "Prepare Protein" dialog box is popped up. Click "Run" after setting is completed, start running.

[0125] Definition of active site: Select ligand of protein, in "Tools Explore", expand Receptor-Ligand Interactions\Define and Edit Binding Sites, click From Current Selection command.

[0126] Docking calculation: open prepared ligand small molecule window and protein window which is prepared and defined active site. In Tools Explore, expand Receptor-Ligand Interaction\Dock Ligands, click Dock Ligands (CDOCKER) command, the corresponding dialog box is popped up. Click Run after setting parameters, run.

[0127] N-(hetero)benzyl substituted tetrahydroquinoline analogue of the application and Hsp90α pharmacophore model molecule docking result: compound I-1 and Hsp90α N-terminal inhibitor cavity produce additional hydrophobic interaction, specific action result see Figure 2 . Compound I-2 and Hsp90α N-terminal inhibitor cavity produce additional π-sulfur interaction, specific action result see Figure 3 . Compound I-3 and Hsp90α N-terminal inhibitor cavity produce additional hydrogen bond interaction, specific action result see Figure 4Structure modified molecule I-4, due to the introduction of methoxy group at para position of benzyl benzene ring, this compound can not only produce hydrophobic or aromatic interaction with pharmacophore model, but also induce additional hydrogen bond interaction, the specific effect results are shown in Figure 5 , which also explains the reason why I-4 has significantly higher HIF-1 inhibitory activity than I-1. Compound I-5 produces additional hydrogen bond and hydrophobic interaction with the cavity of Hsp90α N-terminal inhibitor, the specific effect results are shown in Figure 6 . Compound I-8 produces additional hydrogen bond interaction with the cavity of Hsp90α N-terminal inhibitor, the specific effect results are shown in Figure 7 . Compound I-9 produces additional hydrogen bond interaction with the cavity of Hsp90α N-terminal inhibitor, the specific effect results are shown in Figure 8 . Compound I-12 produces additional hydrophobic interaction with the cavity of Hsp90α N-terminal inhibitor, the specific effect results are shown in Figure 9 . Compound II-2 produces additional hydrophobic interaction with the cavity of Hsp90α N-terminal inhibitor, the specific effect results are shown in Figure 10 .

[0128] Those skilled in the art will readily understand that the above description is only preferred examples of the present application, and is not intended to limit the present application, any modification, equivalent replacement and improvement within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A class of compounds characterized by, The structure of the compound is: 。 2. Use of a compound according to claim 1, characterized in that: The compound I-2, I-3, I-4, I-8, I-9, I-10, I-11 is applied to the preparation of an antitumor drug.

3. Use of a compound according to claim 1, characterized in that: The compound I-3, I-4, I-8, I-9, II-2 is applied to the preparation of an active drug for inhibiting HIF-1 signal pathway.

4. Use of a compound according to claim 1, characterized in that: The compound I-4 is applied to the preparation of an Hsp90 inhibiting drug.

Images