CN106866648B - Phthalimide-type indoleamine-2,3-dioxygenase 1 inhibitors and uses thereof - Google Patents

Abstract

The invention relates to the field of medicinal chemistry, in particular to an IDO1 inhibitor (I) with a phthalimide structure and a preparation method, wherein the definitions of X and R are the same as those in the description. Pharmacodynamic tests prove that the compound of the present invention can be used to treat diseases with pathological characteristics of tryptophan metabolic pathway mediated by IDO1, such as malignant tumors, autoimmune diseases, Alzheimer's disease and schizophrenia.

Description

Phthalimide-type indoleamine-2,3-dioxygenase 1 inhibitors and uses thereof

technical field

The invention relates to the field of medicinal chemistry, in particular to an IDO1 inhibitor with a phthalimide structure and a preparation method.

Background technique

Indoleamine 2,3-dioxygenase (IDO) is a metabolic enzyme containing heme in cells, which is the key rate-limiting enzyme mediating tryptophan metabolism , is an important component of the kynurenine pathway. Since IDO was discovered in 2003, it has been extensively studied by the academic and pharmaceutical circles as an important target for drug development.

The first step in the degradation of tryptophan is the oxidation of tryptophan to N-formyl-L-kynurenine, which is produced by IDO and tryptophan-2,3-dioxygenase (tryptophan 2,3- Dioxygenase, TDO) is mediated by one of the two heme-dependent dioxygenases. Among the two dioxygenases, IDO is considered to be a checkpoint that regulates the oxidation reaction, and its existence and function are the main reasons for the human body to produce immunosuppression. IDO is further divided into IDO1 and IDO2. Under normal circumstances, IDO1 is expressed at a low level in the body, when interferon (IFN-α, IFN-β and IFN-γ), interleukin (IL-1 and IL-2), tumor necrosis factor (TNF) When the factor-induced IDO1 level increases, tryptophan will be metabolized in large quantities, thereby inhibiting the body's immune response to parasitic, viral, bacterial, fungal and other pathogens, and the human body will be in a morbid state of immunosuppression.

The high expression of IDO1 in most tumor cells reduces the concentration of tryptophan in the cell microenvironment, making the synthesis of tryptophan-dependent T cells stagnate in the G1 phase, and the proliferation of T cells is inhibited, thereby inhibiting the immune system of the human body against Killing effect on tumor tissue. At the same time, cytotoxic tryptophan metabolites can have a direct lytic effect on T cells. Therefore, IDO1 plays an important role in tumor immune immunity and tumor development.

In some chronic diseases such as acquired immunodeficiency syndrome (AIDS), various types of depression, Alzheimer's disease, etc., IDO1 is also considered to be one of the reasons for promoting the development of the disease. High levels of interferon induce high expression of IDO1. Under sustained interferon activation, IDO1 reduces the availability of free serum tryptophan, thereby reducing serotonin production. Combined with the accumulation of neuroactive kynurenine metabolites, a variety of neurological and psychological disorders are on the horizon.

Since IDO1 has been proven to be closely related to the pathogenesis of various diseases, IDO1 inhibitors can be used to treat diseases characterized by the pathology of IDO1-mediated tryptophan metabolic pathways, including but not limited to malignant tumors, autoimmune diseases , Alzheimer's disease and schizophrenia. IDO1 inhibitors have broad development prospects as drugs, but so far no suitable IDO1 inhibitors have been marketed as drugs, so finding new and efficient IDO1 inhibitors has important theoretical significance and application value.

Contents of the invention

The invention discloses a compound containing a phthalimide structure, the structural formula is as follows:

X represents hydrogen, halogen or methyl;

R stands for -NH ₂ ,

n stands for 0~3;

m stands for 0 or 1.

wherein X preferably represents methyl.

R is preferred to represent

The preferred partial compounds of the present invention are as follows:

The present invention also discloses a preparation method of compound (I), comprising:

Wherein X, R are as defined above.

The compound represented by formula II reacts with NaNO ₂ to generate the compound represented by formula III. The reaction temperature is preferably 0-5°C, the reaction time is preferably 12-24 hours, and the reaction solvent is a mixed solvent composed of concentrated hydrochloric acid and acetic acid. NaCl was added to the reaction.

The compound represented by formula III reacts with the amine containing R group to generate the compound represented by formula IV. The reaction temperature is preferably 25-30° C., the reaction time is preferably 1-3 h, and the reaction solvent is ethyl acetate, dichloromethane, etc. Inorganic bases or organic bases, such as sodium carbonate, sodium bicarbonate, triethylamine, etc., are also added in the reaction.

The compound represented by formula IV is refluxed to form the compound represented by formula V. The reaction temperature is preferably 80-100° C., the reaction time is preferably 12-20 h, and the reaction solvent is water, ethanol, etc. Inorganic bases such as sodium hydroxide and potassium hydroxide are also added in the reaction.

The general formula shown in Formula I can be generated by refluxing the compound shown in Formula V and X-substituted phthalic anhydride. The reaction temperature is preferably 110-120°C, the reaction time is preferably 12-20h, and the reaction solvent is acetic acid, concentrated hydrochloric acid, and trifluoroacetic acid. Wait.

The compound of general formula I can be purified by common separation methods, such as recrystallization, column chromatography and the like.

The present invention also includes pharmaceutically acceptable salts of the compounds of general formula I and the like.

The compounds of the present invention can be prepared into common pharmaceutical preparations by adding pharmaceutically acceptable carriers, such as tablets, capsules, powders, syrups, liquids, suspensions, injections, spices, sweeteners, liquids or Common pharmaceutical excipients such as solid fillers or diluents.

The clinical administration of the compound of the present invention can be oral administration, injection and the like.

Generally, when the compound of the present invention is used for treatment, the dosage for human is in the range of 1 mg to 1000 mg/day. Depending on the dosage form and the severity of the disease, the dosage may exceed this range.

The following are pharmacological tests and results of some compounds of the present invention:

(1) Determination of IDO1 inhibitory activity

Human indoleamine 2,3-dioxygenase (IDO) with an N-terminal His tag was expressed in Escherichia coli, and IDO1 was extracted and purified. IDO1 catalyzes the oxidative cleavage of the pyrrole ring of the indole ring of tryptophan to give N'-formylkynurenine. 50 mM potassium phosphate buffer (pH 6.5), 40 mM vitamin C, 400 μg/mL catalase, 20 μM methylene blue and IDO1 enzyme were mixed on a 96-well plate. Add the substrate L-tryptophan and the sample to be tested into the above mixed solution. The reaction was carried out at 37° C. for 60 min, and 30% (w/v) trichloroacetic acid was added to terminate the reaction. The 96-well plate was heated at 65° C. for 15 minutes to complete the conversion from formylkynurenine to kynurenine, and then rotated at 6000 rpm for 10 minutes. Take 100 μL of supernatant from each well and transfer it to a new 96-well plate, add 100 μL of acetic acid solution of 2% (w/v) p-(dimethylamino)benzaldehyde, and kynurenine will react with it to produce a yellow color. Use a microplate reader to observe at 480nm, and the obtained results are calculated using IC ₅₀ (nM) calculation software. Using the above method, the IDO1 inhibitory activity of the compound was determined, and its IC ₅₀ (nM) was as follows, and the IDO inhibitor NLG-919 currently in phase I clinical trial was used as a control. The results are shown in Table 1.

Table 1 The inhibitory activity IC ₅₀ (nM) of some compounds of the present invention to IDO1

It can be seen from Table 1 that some compounds of the present invention have good inhibitory activity on IDO1.

(2) Determination of IDO1 inhibitory activity based on Hela cells

At 37 °C, cells were kept in a humidified incubator supplied with 5% _CO2 . The assay was performed as follows: Hela cells were seeded in a 96-well culture plate at a density of 5000/well and cultured overnight. The next day, IFN-γ (final concentration 50 ng/mL) and serial dilutions of the compound (total volume 200 μL medium) were added to the cells. After incubation for 48 h, 140 μL of supernatant/well was transferred to a new 96-well plate. Mix 10 μL of 6.1N trichloroacetic acid into each well, and incubate at 50°C for 30 minutes to hydrolyze the generated N-formylkynurenine into kynurenine. The reaction mixture was then centrifuged at 6000 rpm for 10 min to remove the precipitate. 100 μL of supernatant/well was transferred to another 96-well plate, mixed with 100 μL of 2% (w/v) p-dimethylaminobenzaldehyde in acetic acid, and the yellow color produced by kynurenine was measured at 480 nm. L-kynurenine was used as the standard. Standard solutions (240, 120, 60, 30, 15, 7.5, 3.75, 1.87 μM) were prepared in 100 μL of medium and mixed with an equal volume of 2% (w/v) p-dimethylaminobenzaldehyde. The percentage of inhibition at each concentration was measured, and the data was analyzed using nonlinear regression to obtain the IC ₅₀ (μM) value, and the IDO inhibitor NLG-919 currently in Phase I clinical trials was used as a control. The results are shown in Table 2.

Table 2 The IDO1 inhibitory activity IC ₅₀ (μM) of some compounds of the present invention based on Hela cells

It can be seen from Table 2 that some compounds of the present invention have good inhibitory activity on IDO1.

(3) In vivo experiments on the antitumor activity of IDO1 inhibitors

Murine melanoma cells B16F10 in the vigorous growth stage were collected, prepared into a cell suspension under sterile conditions, and inoculated in the armpit of C57BL6 mice. The transplanted tumors of C57BL6 mice were measured with a vernier caliper to measure the diameter of the transplanted tumors. After the tumors grew to a certain size, the animals were divided into groups with 5 animals in each group. Use the method of measuring tumor diameter to dynamically observe the anti-tumor effect of the test substance. The blank control group was given an equal dose of 0.5% CMC (sodium carboxymethyl cellulose) by intragastric administration; the compound group was given intraperitoneal injection once every other day for 15 days. The long diameter (a) and short diameter (b) of the tumor were measured from the day of administration, and measured every other day, tumor volume = ab ² /2. The tumor-bearing C57BL6 mice were sacrificed 15 days later, and the tumor mass was isolated and weighed. The obtained data were statistically processed, and the tumor inhibition rate was calculated, and the IDO inhibitor NLG-919 currently in Phase I clinical trials was used as a control. The results are shown in Table 3, Figure 1 and Figure 2.

The growth inhibition rate of the B16F10 xenograft tumor of table 3 representative compound CPUL-I021 of the present invention

It can be seen from Figure 1, Figure 2 and Table 3 that the compounds of the present invention have good inhibitory activity on the growth of malignant tumors.

(4) Pharmacokinetic experiments of IDO1 inhibitors in rats

Select 10 SD rats, male, weighing 180-200 g, and give the compound CPUL-I021 by intragastric or intravenous administration, see Table 4:

The administration situation of table 4 compound CPUL-I021 in rats

It is prepared as a suspension with 1% MC for intragastric administration, and as a suspension with DMSO:30% PEG400 (5:95) for intravenous injection. Fasting 12h before the test, free to drink water. Eat uniformly 2 hours after administration. After administration, blood was collected according to the following time points: the blood collection time points of the intragastric administration group were 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24h after administration; the blood collection time points of the intravenous administration group were 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24h. At the above time points, 0.3 mL of venous blood was collected from the retroocular venous plexus of rats, placed in EDTA-2K test tubes, centrifuged at 11,000 rpm for 5 min, separated from plasma, and frozen in a -20°C refrigerator. The concentration of compound CPUL-I021 in rat plasma was determined by LC/MS/MS method. Pharmacokinetic parameters after administration were calculated using non-compartmental models. The peak concentration C _max and peak time T _max are measured values; the area under the drug time curve AUC _0-t value is calculated by the trapezoidal method; AUC _0-∞ = AUC _0-t + C _t /k _e , Ct is the last one The blood drug concentration at the time point can be measured, k _e is the elimination rate constant; the elimination half-life t _1/2 = 0.693/ _ke ; the average residence time MRT = AUMC/AUC. Clearance rate CL=D/AUC _0-∞ ; steady-state volume of distribution V _ss =CL×MRT; absolute bioavailability F=(AUC _gavage ×D _intravenous )/(AUC _intravenous ×D _gavage )×100%. The results are shown in Table 5 and Table 6.

Table 5 Pharmacokinetic parameters after intragastric administration of 10mg/kg CPUL-I021 in rats

Table 6 Pharmacokinetic parameters after intravenous administration of 1mg/kg CPUL-I021 to rats

Description of drawings

Figure 1 is the weighing of isolated tumor mass after sacrificing tumor-bearing C57BL6 mice

Figure 2 is the relationship between tumor volume and days of administration

Detailed ways

Example 1

Preparation of 4-amino-3-(N'-hydroxyamidino)-1,2,5-oxadiazole(II)

Add sodium nitrite (20.7g, 300mmol) and 40mL water into a 500mL reaction flask, stir to dissolve, slowly add malononitrile (9.9g, 150mmol) hydrochloric acid (2N) solution dropwise at room temperature, stir overnight at room temperature, and cool to 0°C, add dropwise an aqueous solution of hydroxylamine hydrochloride (23g, 340mmol), stir for 30min, adjust the pH to 10 with 10N sodium hydroxide below 20°C, rise to 35°C for 2h, and then heat to reflux for 2h. Cool to room temperature, extract the reaction solution with 20 mL of ethyl acetate, distill off the solvent under reduced pressure to obtain a white solid, place the water layer overnight, precipitate a solid, filter with suction, wash with water, combine the filter cake with the white solid obtained by distillation under reduced pressure and dry to obtain a white powder like solid (14.9 g, 69.5%). ¹ H NMR (300MHz,DMSO-d ₆ )δ:10.46(s,1H,-OH),6.24(s,2H,-NH ₂ ),6.02(s,2H,-NH ₂ )ppm.HRMS m/z [M+H] ⁺ calculated for C ₃ H ₅ N ₅ O ₂ : 143.0516, found: 143.0518.

Preparation of 4-Amino-3-Chloromethoximino-1,2,5-oxadiazole(III)

Add II (4.18g, 29.2mmol), 58.4mL of water, 29.7mL of acetic acid and 14.6mL of 6N hydrochloric acid into a 250mL reaction flask, rise to 45°C and stir to dissolve, add solid sodium chloride (5.12g, 87.5mmol), Stir to dissolve, cool to 0°C, a white solid precipitates, slowly add an aqueous solution of sodium nitrite (2.00g, 29mmol) dropwise, stir for 3 hours below 0°C, rise to room temperature, filter with suction, wash the filter cake with water, and recrystallize from toluene to obtain a white powder like solid (2.52 g, 53.2%). ¹ H NMR(300MHz,DMSO-d ₆ )δ:13.39(s,1H,-OH),6.29(s,2H,-NH ₂ )ppm.HRMS m/z[M+H] ⁺ calculated for C ₃ H ₃ ClN ₄ O ₂ : 161.9872, found: 161.9872.

Example 2

Preparation of Compound CPUL-I001

Preparation of 4-amino-3-(N-methyl-N'-hydroxyamidino)-1,2,5-oxadiazole (IV-1)

Add III (1.0g, 6.2mmol) and 40mL ethanol into a 100mL reaction flask, stir, and slowly add 40% methylamine ethanol solution (0.48g, 6.2mmol) and triethylamine (1.25g, 12.4 mmol), stirred at room temperature for 3 h, evaporated the solvent under reduced pressure, added ethyl acetate and stirred, filtered with suction, washed the filtrate once with water and saturated aqueous sodium chloride solution, dried the organic layer with anhydrous sodium sulfate, evaporated the solvent under reduced pressure to obtain yellow Oil (0.68g, 70.1%). HRMS m/z [M+H] ⁺ calculated for C ₄ H ₇ N ₅ O ₂ : 157.0602, found: 157.0600.

Preparation of 4-methylamino-3-(N'-hydroxyamidino)-1,2,5-oxadiazole (V-1)

Add IV-1 (0.68g, 4.3mmol) and 9mL water into a 50mL reaction flask, slowly add 3mL aqueous solution of potassium hydroxide (0.73g, 13mmol) dropwise under stirring, heat and reflux for 14h, after the reaction is detected by TLC, Cool to room temperature, extract with ethyl acetate (5×4 mL), combine the organic phases, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure to obtain a light yellow oil (0.46 g, 67.5%). HRMS m/z [M+H] ⁺ calculated for C ₄ H ₇ N ₅ O ₂ : 157.0608, found: 157.0600.

Preparation of 2-[(4-methylamino-1,2,5-oxadiazol-3-yl)methoximino]isoindole-1,3-dione (CPUL-I001)

Add V-1 (0.46g, 3mmol), 20mL acetic acid and phthalic anhydride (0.88g, 6mmol) into a 100mL reaction flask, reflux and stir for about 8h, after the reaction is detected by TLC, cool to room temperature, and depressurize The solvent was evaporated and purified by column chromatography to obtain a pale yellow solid (0.25 g, 29%). mp203-204℃, ¹ H NMR(300MHz,DMSO-d ₆ )δ:12.61(s,1H,-OH),7.87-7.95(m,2H,-ArH),7.64-7.67(m,2H,-ArH ),5.41(m,1H,-NH-),2.33(d,J＝4.5Hz,3H,-CH ₃ )ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₂ H ₉ N ₅ O ₄ :287.0650,found:287.0655.

Example 3

Preparation of compound CPUL-I002

Compound CPUL-I002 (0.28 g, 15%) was synthesized in the same manner as compound CPUL-I001 except that ethylamine (6.2 mmol) was used instead of methylamine. mp199-200℃, ¹ H NMR (300MHz,DMSO-d ₆ )δ:13.01(s,1H,-OH),7.91-8.02(m,2H,-ArH),7.73-7.77(m,2H,-ArH ),6.21(m,1H,-NH-),2.43(m,2H,-CH ₂ -),2.29(t,J=4.7Hz,3H,-CH ₃ )ppm.HRMS m/z[M+H ] ⁺ calculated for C ₁₂ H ₉ N ₅ O ₄ : 301.0806, found: 301.0811.

Example 4

Preparation of compound CPUL-I003

Compound CPUL-I003 (0.28 g, 14.5%) was synthesized in the same manner as compound CPUL-I001 except that cyclopropylamine (6.2 mmol) was used instead of methylamine. mp169-171℃, ¹ H NMR(300MHz,DMSO-d ₆ )δ:13.71(s,1H,-OH),7.81-8.03(m,4H,-ArH),6.63(bs,1H,-NH-) ,2.67-2.74(m,1H,-CH-),0.67-0.78(m,4H,-CH ₂ -CH ₂ -)ppm. HRMS m/z[M+H] ⁺ calculated for C ₁₄ H ₁₁ N ₅ O ₄ : 313.0884,found: 313.0867.

Example 5

Preparation of Compound CPUL-I004

Compound CPUL-I004 (0.31 g, 16%) was synthesized in the same manner as compound CPUL-I001 except that n-propylamine (6.2 mmol) was used instead of methylamine. mp197-198℃, ¹ H NMR (300MHz,DMSO-d ₆ )δ:13.61(s,1H,-OH),7.97-8.05(m,2H,-ArH),7.84-7.87(m,2H,-ArH ), 6.41 (bs, 1H, -NH-), 3.28 (q, J = 6.5Hz, 2H, -CH ₂ -), 1.63-1.70 (m, 2H, -CH ₂ -), 0.92 (t, J = 7.3Hz, 3H, -CH ₃ )ppm. HRMS m/z[M+H] ⁺ calculated for C ₁₄ H ₁₃ N ₅ O ₄ : 315.0873, found: 315.0869.

Example 6

Preparation of Compound CPUL-I005

Compound CPUL-I005 (0.32 g, 15.5%) was synthesized in the same manner as compound CPUL-I001 except that tert-butylamine (6.2 mmol) was used instead of methylamine. mp136-138℃, ¹ H NMR (300MHz,DMSO-d ₆ )δ:13.67(s,1H,-OH),7.85-7.97(m,4H,-ArH),5.80(s,1H,-NH-) ,1.40(s,9H,-C(CH ₃ ) ₃ )ppm. HRMS m/z[M+H] ⁺ calculated for C ₁₅ H ₁₅ N ₅ O ₄ : 329.1197, found: 329.1196.

Example 7

Preparation of compound CPUL-I006

Compound CPUL-I006 (0.32 g, 14.5%) was synthesized in the same manner as compound CPUL-I001 except that cyclohexylamine (6.2 mmol) was used instead of methylamine. mp202-203℃, ¹ H NMR (300MHz, (CD ₃ ) ₂ CO-d ₆ )δ:8.14-8.17(m,1H,-ArH),7.98-8.01(m,1H,-ArH),7.88-7.90 (m,2H,-ArH),5.71(d,J=6.9Hz,1H,-NH-),3.55(bs,1H,-CH-),2.11-2.17(m,2H,-CH ₂ -), 1.77-1.80(m,2H,-CH ₂ -),1.31-1.68(m,6H,-CH ₂ -CH ₂ -CH ₂ -)ppm. HRMS m/z[M+H] ⁺ calculated for C ₁₇ H ₁₇ N ₅ O ₄ : 355.1353, found: 355.1353.

Example 8

Preparation of compound CPUL-I007

Compound CPUL-I007 (0.28 g, 13%) was synthesized in the same manner as compound CPUL-I001 except that 4-oxazacyclohexane (6.2 mmol) was used instead of methylamine. mp190-192℃, ¹ H NMR (300MHz,DMSO-d ₆ )δ:13.72(s,1H,-OH),7.81-8.06(m,4H,-ArH),3.64-3.74(m,4H,-CH ₂ -O-CH ₂ -),3.31(t,J＝4.2Hz,4H,-CH ₂ -N-CH ₂ -)ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₅ H ₁₃ N ₅ O ₅ : 343.0989, found: 343.0992.

Example 9

Preparation of compound CPUL-I008

Compound CPUL-I008 (0.32 g, 15%) was synthesized in the same manner as compound CPUL-I001 except that aniline (6.2 mmol) was used instead of methylamine. mp182-184℃, ¹ H NMR (300MHz,DMSO-d ₆ )δ:8.69(s,1H,-NH-),7.98-8.06(m,2H,-ArH),7.84-7.89(m,2H,- ¹ H RMS m/z [M+H] ⁺ calculated for C ₁₇ H ₁₁ N ₅ O ₄ : 349.0884, found: 349.0916.

Example 10

Preparation of Compound CPUL-I009

Compound CPUL-I009 (0.35 g, 15.5%) was synthesized in the same manner as compound CPUL-I001 except that benzylamine (6.2 mmol) was used instead of methylamine. mp195-197℃, ¹ H NMR (300MHz, DMSO-d ₆ )δ:7.97-8.06(m,2H,-ArH),7.82-7.89(m,2H,-ArH),7.24-7.43(m,5H, -ArH),7.10(t,J=6.2Hz,1H,-NH-),4.53(d,J=6.1Hz,2H,-CH ₂ -)ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₈ H ₁₃ N ₅ O ₄ : 363.1040, found: 363.1045.

Example 11

Preparation of Compound CPUL-I010

Compound CPUL-I010 (0.31 g, 14.5%) was synthesized in the same manner as compound CPUL-I001, except that propylamine (6.2 mmol) was used instead of methylamine and 4-chlorophthalic anhydride was used instead of phthalic anhydride. mp120-122℃, ¹ H NMR(300MHz,DMSO-d ₆ )δ:14.00(s,1H,-OH),7.90-8.09(m,3H,-ArH),6.42(bs,1H,-NH-) ,3.26(bs,2H,-CH ₂ -),1.63-1.67(m,2H,-CH ₂ -),0.91(bs,3H,-CH ₃ )ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₄ H ₁₂ ClN ₅ O ₄ : 349.0651, found: 349.0686.

Example 12

Preparation of compound CPUL-I011

Compound CPUL-I011 (0.41 g, 17%) was synthesized in the same manner as compound CPUL-I001, except that methylamine was replaced by cyclohexylamine (6.2 mmol) and phthalic anhydride was replaced by 4-chlorophthalic anhydride. mp200-202℃, ¹ H NMR(300MHz,DMSO-d ₆ )δ:14.00(s,1H,-OH),8.03-8.06(m,2H,-ArH),7.93-7.96(m,1H,-ArH ), 5.96(d, J=7.6Hz, 1H, -NH-), 3.31-3.43(m, 1H, -CH-), 2.00-2.04(m, 2H, -CH ₂ -), 1.71-1.75(m ,2H,-CH ₂ -),1.31-1.63(m,6H,-CH ₂ -CH ₂ -CH ₂ -)ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₇ H ₁₆ ClN ₅ O ₄ :389.0964,found:389.0978.

Example 13

Preparation of compound CPUL-I012

Compound CPUL-I012 (0.39 g, 16.5%) was synthesized in the same manner as compound CPUL-I001, except that methylamine was replaced by 4-oxazacyclohexane (6.2 mmol) and phthalic anhydride was replaced by 4-chlorophthalic anhydride. mp176-178℃, ¹ H NMR (300MHz,DMSO-d ₆ )δ:13.90(s,1H,-OH),7.93-8.08(m,3H,-ArH),3.73(bs,4H,-CH ₂ - O-CH ₂ -),3.31(bs,4H,-CH ₂ -N-CH ₂ -)ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₅ H ₁₂ ClN ₅ O ₅ :377.0600,found: 377.0600.

Example 14

Preparation of compound CPUL-I013

Compound CPUL-I013 (0.34 g, 14.5%) was synthesized in the same manner as compound CPUL-I001, except that methylamine was replaced by aniline (6.2 mmol) and phthalic anhydride was replaced by 4-chlorophthalic anhydride. mp207-209℃, ¹ H NMR (300MHz, DMSO-d ₆ )δ: 8.77(s, 1H, -NH-), 8.06(s, 1H, -ArH), 7.95(d, J＝7.5Hz, 1H, -ArH), 7.88(d, J=6.8Hz, 1H, -ArH), 7.67(d, J=7.8Hz, 2H, -ArH), 7.39(t, J=7.6Hz, 2H, -ArH), 7.06 (t, J=7.1 Hz, 1H, -ArH) ppm. HRMS m/z [M+H] ⁺ calculated for C ₁₇ H ₁₀ ClN ₅ O ₄ : 383.0494, found: 383.0512.

Example 15

Preparation of compound CPUL-I014

Compound CPUL-I014 (0.38 g, 15.5%) was synthesized in the same manner as compound CPUL-I001, except that propylamine (6.2 mmol) was used instead of methylamine and 4-bromophthalic anhydride was used instead of phthalic anhydride. mp130-132℃, ¹ H NMR (300MHz, DMSO-d ₆ )δ:14.00(s,1H,-OH),7.93-8.21(m,3H,-ArH),6.43(t,J＝5.3Hz,1H ,-NH-),3.24-3.31(m,2H,-CH ₂ -),1.63-1.70(m,2H,-CH ₂ -),0.92(t,J＝7.4Hz,3H,-CH ₃ )ppm .HRMS m/z [M+H] ⁺ calculated for C ₁₄ H ₁₂ BrN ₅ O ₄ : 393.0145, found: 393.0148.

Example 16

Preparation of Compound CPUL-I015

Compound CPUL-I015 (0.46 g, 17%) was synthesized in the same manner as compound CPUL-I001, except that cyclohexylamine (6.2 mmol) was used instead of methylamine and 4-bromophthalic anhydride was used instead of phthalic anhydride. mp188-190℃, ¹ H NMR (300MHz, CDCl ₃ )δ:7.77-8.23(m,3H,-ArH),5.30(bs,1H,-NH-),3.58(bs,1H,-CH-), 2.15-2.21(m,2H,-CH ₂ -),1.70-1.78(m,2H,-CH ₂ -),1.25-1.66(m,6H,-CH ₂ -CH ₂ -CH ₂ -)ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₇ H ₁₆ BrN ₅ O ₄ : 433.0458, found: 433.0462.

Example 17

Preparation of compound CPUL-I016

Compound CPUL-I016 (0.39 g, 15%) was synthesized in the same manner as compound CPUL-I001, except that methylamine was replaced by 4-oxazacyclohexane (6.2 mmol) and phthalic anhydride was replaced by 4-bromophthalic anhydride. mp189-191℃, ¹ H NMR (300MHz, DMSO-d ₆ )δ: 13.95(s, 1H, -OH), 7.91-8.21(m, 3H, -ArH), 3.73(t, J＝4.2Hz, 4H ,-CH ₂ -O-CH ₂ -),3.31(t,J=4.1Hz,4H,-CH ₂ -N-CH ₂ -)ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₅ H ₁₂ BrN ₅ O ₅ : 421.0095, found: 421.0098.

Example 18

Preparation of compound CPUL-I017

Compound CPUL-I017 (0.40 g, 15%) was synthesized in the same manner as compound CPUL-I001, except that methylamine was replaced by aniline (6.2 mmol) and phthalic anhydride was replaced by 4-bromophthalic anhydride. mp237-239℃, ¹ H NMR (300MHz, DMSO-d ₆ )δ:8.69(s,1H,-NH-),7.95-8.06(m,3H,-ArH),7.68(d,J＝7.9Hz, 2H, -ArH), 7.40(t, J=7.8Hz, 2H, -ArH), 7.07(t, J=7.3Hz, 1H, -ArH) ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₇ H ₁₀ BrN ₅ O ₄ : 426.9989, found: 427.0004.

Example 19

Preparation of Compound CPUL-I018

Compound CPUL-I018 (0.29 g, 14%) was synthesized in the same manner as compound CPUL-I001, except that propylamine (6.2 mmol) was used instead of methylamine and 4-methylphthalic anhydride was used instead of phthalic anhydride. mp123-125℃, ¹ H NMR (300MHz, CDCl ₃ )δ: 7.68-8.08(m,2H,-ArH),7.57(d,J＝7.8Hz,1H,-ArH),5.46(bs,1H,- NH-), 3.40-3.43(m, 2H, -CH ₂ -), 2.54(s, 3H, -CH ₃ ), 1.74-1.81(m, 2H, -CH ₂ -), 1.03(t, J=7.4 Hz, 3H, -CH ₃ ) ppm. HRMS m/z[M+H] ⁺ calculated for C ₁₅ H ₁₅ N ₅ O ₄ : 329.1197, found: 329.1205.

Example 20

Preparation of Compound CPUL-I019

Compound CPUL-I019 (0.33 g, 14.5%) was synthesized in the same manner as compound CPUL-I001 except that cyclohexylamine (6.2 mmol) was used instead of methylamine and 4-methylphthalic anhydride was used instead of phthalic anhydride. mp201-203℃, ¹ H NMR (300MHz, CDCl ₃ ),δ:7.89-8.05(m,1H,-ArH),7.66-7.81(m,1H,-ArH),7.55(d,J＝7.9Hz, 1H, -ArH), 5.36(d, J=5.6Hz, 1H, -NH-), 3.57(bs, 1H, -CH-), 2.53(s, 3H, -CH ₃ ), 2.15-2.18(m, 2H,-CH ₂ -),1.69-1.77(m,2H,-CH ₂ -),1.24-1.65(m,6H,-CH ₂ -CH ₂ -CH ₂ -)ppm.HRMS m/z[M+ H] ⁺ calculated for C ₁₈ H ₁₉ N ₅ O ₄ : 369.1510, found: 369.1507.

Example 21

Preparation of Compound CPUL-I020

Compound CPUL-I020 (0.33g, 15.5%) was synthesized in the same way as compound CPUL-I001, except that methylamine was replaced by 4-oxazacyclohexane (6.2mmol) and phthalic anhydride was replaced by 4-methylphthalic anhydride. . mp176-178℃, ¹ H NMR (300MHz, DMSO-d ₆ )δ: 13.58(s, 1H, -OH), 7.74-7.96(m, 3H, -ArH), 3.73(t, J＝4.8Hz, 4H ,-CH ₂ -O-CH ₂ -), 3.31(t,J=4.4Hz,4H,-CH ₂ -N-CH ₂ -),2.49(s,3H,-CH ₃ )ppm.HRMS m/z [M+H] ⁺ calculated for C ₁₆ H ₁₅ N ₅ O ₅ : 357.1146, found: 357.1150.

Example 22

Preparation of Compound CPUL-I021

Compound CPUL-I021 (0.36 g, 16%) was synthesized in the same manner as compound CPUL-I001, except that methylamine was replaced by aniline (6.2 mmol) and phthalic anhydride was replaced by 4-methylphthalic anhydride. mp206-208℃, ¹ H NMR (300MHz, (CD ₃ ) ₂ CO-d ₆ )δ: 12.58(s, 1H, -OH), 8.38(s, 1H, -NH-), 8.03(dd, J= 8.0Hz, 5.4Hz, 1H, -ArH), 7.88(d, J＝8.4Hz, 1H, -ArH), 7.71-7.75(m, 3H, -ArH), 7.44(t, J＝8.1Hz, 2H, -ArH),7.11(t,J＝7.3Hz,1H,-ArH),2.57(s,3H,-CH ₃ )ppm.HRMS m/z[M+H] ⁺ calculated for C ₁₈ H ₁₃ N ₅ O ₄ :363.1040,found:363.1041.

Claims (7)

1. A compound of general formula I or a pharmaceutically acceptable salt thereof: X represents hydrogen, halogen or methyl; R stands for -NH ₂ , n stands for 0~3; m stands for 0 or 1. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein X represents methyl. 3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R represents 4. The preparation method of the compound of claim 1,2 or 3, comprising: Wherein the definition of X, R is the same as claim 1. 5. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 6. Use of the compound of claim 1, 2 or 3 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating diseases characterized by the pathology of the IDO1-mediated tryptophan metabolic pathway. 7. The use according to claim 6, wherein the disease characterized by the pathology of the IDO1-mediated tryptophan metabolic pathway is a malignant tumor.