CN114835621A

CN114835621A - Imine IDO inhibitor and preparation and application thereof

Info

Publication number: CN114835621A
Application number: CN202110131014.7A
Authority: CN
Inventors: 杨金玉; 刘文丽; 赵力挥; 唐海
Original assignee: Jiangsu Tasly Diyi Pharmaceutical Co Ltd
Current assignee: Jiangsu Tasly Diyi Pharmaceutical Co Ltd
Priority date: 2021-01-30
Filing date: 2021-01-30
Publication date: 2022-08-02

Abstract

The invention relates to imine IDO inhibitors, and preparation and therapeutic application thereof, wherein the imine IDO inhibitors comprise imine compounds shown in formula I or pharmaceutically acceptable salts thereof, which can be used as antitumor immunotherapy medicaments, and substituent groups shown in formula IAs described in the specification, the IDO inhibitors of the present invention are characterized by: excellent activity at cellular level as IDO inhibitor, IC thereof ₅₀ Less than 1. mu.M.

Description

Imine IDO inhibitor and preparation and application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to an imine compound and a preparation method thereof, wherein the imine compound is used as a therapeutic drug, particularly as an application of the imine compound in tumor immunotherapy by inhibiting tryptophan metabolism, and related biomarkers include but are not limited to IDO1 (indoleamine 2,3-dioxygenase 1).

Background

By the year 2030, the number of cancer patients worldwide will reach 2400 ten thousand as estimated by the international agency for research on cancer (IARC). In recent years, immunotherapy has become a major focus in precise cancer therapy, and has gradually progressed to a fourth tumor treatment modality following surgery, chemotherapy, and radiotherapy. Indoleamine 2,3-dioxygenase (IDO) is a monomeric protein containing iron heme, takes superoxide anion as a cofactor, catalyzes the epoxylytic cleavage of L-tryptophan indole, and is distributed in tissues and cells outside the liver of human and other mammals. IDO1 plays an important regulatory role in inhibiting T cell immunity and inducing tumor immune tolerance. In healthy humans, IDO1 expression levels were low, but when the organism was in infection, inflammation, IDO1 expression was significantly increased, especially in various tumor tissues. Tumor cells can recruit IDO 1-expressing dendritic cells into a tumor microenvironment, so that IDO1 is highly expressed in various tumor tissues and draining lymph nodes of human beings, such as ovarian cancer, lung cancer, chronic lymphocytes, leukemia and the like.

IDO has become a hot target for tumor immunotherapy. The IDO inhibitors of various small molecular compounds are combined with antibody immune medicaments in clinical tests to achieve better curative effect on some refractory cancers. Currently, the number of global IDO inhibitors in research exceeds 50, wherein 1 phase III clinical drug, 2 phase II clinical drugs, 7 phase I clinical drugs, and 30 preclinical drugs are available. The research of developing targeted small molecule drugs acting on IDO can obtain the IDO inhibitor with better activity and higher selectivity, and has very important social and economic significance.

IDO inhibitors can be divided into three major classes: competitive with epacadostat as representativeInhibitors, non-competitive inhibitors typified by navoximod, and other classes of inhibitors. Among them the most rapidly progressing are competitive inhibitors. Epacadostat and 1-methyl tryptophan (1-MT) play a role in inhibiting through competitively binding with tryptophan to an IDO catalytic region, thereby effectively intervening in the metabolic process of tryptophan and enabling the proliferation of effector T cells and natural killer cells to be enhanced, the apoptosis to be reduced and CD86 ⁺ Dendritic cell activation is increased.

1-methyl tryptophan (1-MT) is in the 90 s of the 20 th century, and a derivative obtained by performing structural modification on substrate tryptophan of IDO as a template is an IDO inhibitor which is commonly used in vitro and in vivo experiments at present, and the inhibition constant (Ki) of the derivative is 34 uM. 1-MT is of the formula:

chinese patent CN109111438A discloses a series of amidine compounds for IDO inhibitors, however, the inventors have not conducted further activity studies on the series of compounds, and only tested the IDO enzyme inhibitory activity of the compounds of examples 16, 17 and 19, and thus could only demonstrate the extracellular IDO inhibitory effect of the above examples. Since the patent does not report any data on the intracellular or in vivo antiproliferative activity of the examples, these compounds cannot be considered to have antitumor activity in consideration of the large difference between the environment of the activity test described in the patent and the environment in the cell or in vivo.

In order to develop an inhibitor with a new structure, the invention provides a compound with IDO inhibition effect and application thereof. The novel compound of the invention is characterized in that: the compounds are excellent in activity at the cellular level as IDO inhibitors, and their IC ₅₀ Less than 1. mu.M.

Disclosure of Invention

The invention provides a compound which has IDO1 inhibiting activity and can be used as a therapeutic or remission drug for tumors.

Specifically, the invention provides an imine compound shown as a formula I or a pharmaceutically acceptable salt thereof:

wherein:

r1 is hydroxy, substituted or unsubstituted 5-8 membered aryl or heteroaryl;

r2 is cyano, substituted or unsubstituted 5-8 membered arylamino;

w is selected from C, N;

x is selected from C, N;

y is selected from C, N;

z is selected from C, N;

wherein W and X are not N at the same time, and Y and Z are not C at the same time. Preferably, the first and second liquid crystal materials are,

r1 is hydroxy, or substituted 5-8 membered aryl,

r2 is cyano, or a substituted 5-to 8-membered arylamino.

It is still further preferred that the first and second substrates,

r1 is hydroxy, or substituted phenyl,

r2 is cyano, or substituted phenylamino.

It is still further preferred that the concentration of the organic compound,

r1 is hydroxy, or phenyl substituted by halogen,

r2 is cyano, or phenylamino substituted with halogen.

More preferably, it is a mixture of more preferably,

r1 is hydroxy, or

R2 is cyano, or

Most preferably, the first and second substrates are,

the imine compound is selected from:

the pharmaceutically acceptable salt of the imine compound is a salt formed by the imine compound and the following acids: hydrochloric acid, p-toluenesulfonic acid, tartaric acid, maleic acid, lactic acid, methanesulfonic acid, sulfuric acid, phosphoric acid, citric acid, acetic acid or trifluoroacetic acid.

Preferably, the compound is a salt of an imine compound with: p-toluenesulfonic acid, hydrochloric acid, tartaric acid or trifluoroacetic acid.

The pharmaceutically acceptable salts of the imine compounds further comprise solvates, metabolite prodrugs and the like of the imine compounds.

The invention also provides a pharmaceutical composition containing the imine compound or the pharmaceutically acceptable salt thereof.

The pharmaceutical composition of the present invention, preferably in the form of a unit dose pharmaceutical preparation, can be formulated into any pharmaceutically acceptable dosage form selected from the group consisting of: tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, hard capsules, soft capsules, oral liquids, buccal agents, granules, suspensions, solutions, injections, suppositories, ointments, plasters, creams, sprays, patches. Preferred are oral dosage forms, most preferred are tablets, capsules.

Furthermore, the pharmaceutical composition of the invention also contains a pharmaceutically acceptable carrier.

The pharmaceutical preparation can be prepared by conventional techniques in pharmaceutical science, such as mixing the imine compound, or the solvate thereof, or the pharmaceutically acceptable salt of the imine compound, or the pharmaceutically acceptable salt of the solvate thereof, with a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers include, but are not limited to: mannitol, sorbitol, sorbic acid or potassium salt, sodium metabisulfite, sodium bisulfite, sodium thiosulfate, cysteine hydrochloride, thioglycolic acid, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone, disodium EDTA, calcium sodium EDTA, carbonates of monovalent alkali metals, acetates, phosphates or aqueous solutions thereof, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acids, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivatives, cellulose and derivatives thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, propylene glycol, ethanol, Tween 60-80, span-80, beeswax, lanolin, liquid paraffin, cetyl alcohol, gallic acid esters, agar, triethanolamine, basic amino acids, Urea, allantoin, calcium carbonate, calcium bicarbonate, surfactant, polyethylene glycol, cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talcum powder, calcium stearate, magnesium stearate and the like.

When the pharmaceutical composition is prepared into a medicament, the medicament with unit dose can contain 0.1-1000mg of the pharmaceutical active substance, and the balance is a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be 0.1-99.9% by weight of the total weight of the formulation.

The pharmaceutical composition of the present invention is administered in an amount determined according to the condition of the patient.

The invention also comprises the application of the imine compound or the pharmaceutically acceptable salt thereof and a pharmaceutical composition containing the imine compound or the pharmaceutically acceptable salt thereof in preparing a medicament for treating IDO 1-related diseases.

The disease associated with IDO1 includes, but is not limited to, tumors. The types of cancer that can be treated with the compounds of the present invention include, but are not limited to, brain, skin, bladder, ovarian, breast, stomach, pancreatic, prostate, colon, blood, lung, and bone cancers. Examples of such cancer types include neuroblastoma; bowel cancers such as rectal cancer, colon cancer, familial adenomatous polyposis carcinoma, and hereditary nonpolyposis colorectal cancer; esophageal cancer; lip cancer; laryngeal cancer; hypopharyngeal carcinoma; tongue cancer; salivary gland cancer; stomach cancer; adenocarcinoma; medullary thyroid carcinoma; papillary thyroid carcinoma; kidney cancer; renal parenchymal carcinoma; ovarian cancer; cervical cancer; uterine body cancer; endometrial cancer; choriocarcinoma; pancreatic cancer; prostate cancer; testicular cancer; breast cancer; urinary cancer; melanoma; brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral neuroectodermal tumors; hodgkin lymphoma; non-hodgkin's lymphoma; burkitt's lymphoma; acute Lymphocytic Leukemia (ALL); chronic lymphocytic leukemia (AML); chronic Myeloid Leukemia (CML); adult T cell leukemia lymphoma; diffuse large B-cell lymphoma (DLBCL); hepatocellular carcinoma; gallbladder cancer; bronchial cancer; small cell lung cancer; non-small cell lung cancer; multiple myeloma; a basal cell tumor; teratoma; retinoblastoma; choroidal melanoma; seminoma; rhabdomyosarcoma; craniopharyngioma; osteosarcoma; chondrosarcoma; myosarcoma; liposarcoma; fibrosarcoma; ewing's sarcoma and plasmacytoma.

The compound of the invention can be prepared by adopting the following preparation method:

the first scheme is as follows:

in the first scheme, 3-bromo-4-fluoroaniline is used as a raw material and condensed with indole-7-carboxylic acid under an alkaline condition to obtain a compound 1-2, the compound 1-2 is subjected to thionation by a Lawson reagent to obtain a compound 1-3, and the compound 1-3 is reacted with hydroxylamine hydrochloride to obtain a compound 1.

Scheme II:

in the second scheme, 4-indole carbaldehyde is used as a raw material, a compound 2-2 is obtained through BOC protection, a compound 2-3 is obtained through reaction of the compound 2-2 and hydroxylamine hydrochloride, a compound 2-4 is obtained through chlorination of the compound 2-3, a compound 2-5 is obtained through reaction of the compound 2-4 and 3-bromo-4-fluoroaniline, and a compound 2-5 is obtained through deprotection of the compound 2-5.

The third scheme is as follows:

in the third scheme, 4-indole carbaldehyde is used as a raw material, and is condensed with 3-bromo-4-fluoroaniline and cyanated to obtain a compound 3.

And the scheme is as follows:

in the fourth scheme, 7-indole carbaldehyde is used as a raw material, and is condensed with 3-bromo-4-fluoroaniline and cyanated to obtain a compound 4.

And a fifth scheme:

in the fifth scheme, 2-amino-3-nitrobenzoic acid methyl ester is used as a raw material, and is subjected to reduction condensation to obtain a compound 5-2, the compound 5-2 is subjected to reduction and oxidation to obtain a compound 5-4, and the compound 5-4 is subjected to condensation and cyanation with 3-bromo-4-fluoroaniline to obtain a compound 5.

Scheme six:

in the sixth scheme, 7-chloro-1H-pyrrolo [3,2-b ] pyridine is used as a raw material, and is subjected to coupling and esterification by carbon monoxide to obtain a compound 6-2, the compound 6-2 is subjected to reduction and oxidation to obtain a compound 6-4, and the compound 6-4 is subjected to condensation and cyanation with 3-bromo-4-fluoroaniline to obtain a compound 6.

The scheme is seven:

in the seventh scheme, 7-bromo-1H-pyrrolo [3,2-c ] pyridine is used as a raw material, and is subjected to coupling and esterification by carbon monoxide to obtain a compound 7-2, the compound 7-2 is subjected to reduction and oxidation to obtain a compound 7-4, and the compound 7-4 is subjected to condensation and cyanation with 3-bromo-4-fluoroaniline to obtain a compound 7.

Description of the drawings:

FIG. 1 preparation of Compound 1 ¹ H-NMR spectrum

FIG. 2 preparation of Compound 2 ¹ H-NMR spectrum

FIG. 3 preparation of Compound 3 ¹ H-NMR spectrum

FIG. 4 preparation of Compound 4 ¹ H-NMR spectrum

FIG. 5 preparation of Compound 5 ¹ H-NMR spectrum

FIG. 6 preparation of Compound 6 ¹ H-NMR spectrum

FIG. 7 preparation of Compound 7 ¹ H-NMR spectrum

The specific implementation mode is as follows:

example 1: synthesis of Compound 1

a. Synthesis of Compound 1-2

3-bromo-4-fluoroaniline (1.89g,10mmol) and indole-7-carboxylic acid (1.61g,10mmol) were dissolved in 20mL DCM and triethylamine (3.03g,30mmol) and HATU (3.8g,10mmol) were added with stirring. The reaction was stirred at room temperature for 2 h. After the reaction is finished, the reaction solution is decompressed and concentrated, and the crude product is purified by column chromatography to obtain 2.0g of white solid with the yield of 61%.

b. Synthesis of Compounds 1-3

Compound 1-2(1g,3mmol) was dissolved in toluene (10mL) and Lawson's reagent (0.97g,2.4 mmol) was added. The reaction was heated to reflux and the reaction was continued for 14 h. After the reaction is finished, the reaction solution is decompressed and concentrated, and the crude product is purified by column chromatography to obtain 0.62g of white solid with the yield of 52.7%.

c. Synthesis of Compound 1

Under argon, compound 1-3(620g,1.78mmol), hydroxylamine hydrochloride (614mg,8.9mmol) and sodium bicarbonate (1.498g,17.8mmol) were added to 20mL of ethanol. The reaction solution was heated to 70 ℃ and the reaction was continued for 10 h. After the reaction was completed, the reaction mixture was cooled to room temperature. The reaction solution was filtered, and the filter cake was washed with ethyl acetate. The filtrate was collected and concentrated under reduced pressure, and the crude product was purified by column chromatography to give 0.126g of a pale yellow solid in 20% yield.

MS Found:348.2[M+H] ⁺ ,350.2[M+2+H] ⁺ 。

Example 2: synthesis of Compound 2

a. Synthesis of Compound 2-2

Potassium tert-butoxide (2.9g,25.8mmol) and di-tert-butyl dicarbonate (4.5g,20.6 mmol) are added to 4-indolecarboxaldehyde (2.5g,17.2mmol) in 50mL anhydrous THF under nitrogen. The reaction is carried out for 12h at room temperature. After completion of the reaction, the reaction mixture was concentrated and extracted with ethyl acetate (50mL × 3). The organic phases were combined and dried over sodium sulfate. Filtering, concentrating under reduced pressure to obtain crude product, and performing column chromatography (0-10% EA in PE) to obtain white solid 1.7g with yield of 40.3%. MS Found 246.2[ M + H ]] ⁺ 。

b. Synthesis of Compounds 2-3

To a solution of compound 2-2(1.5g,6.1mmol) in ethanol (50mL) under nitrogen was added hydroxylamine hydrochloride (2.1g,30.2 mmol). The reaction solution was heated to 70 ℃ and reacted for 4 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and extracted with ethyl acetate (50mL × 3). The combined organic phases were concentrated and washed with petroleum ether to give 1.2g of an off-white solid in 75.6% yield. MS Found 261.2[ M + H] ⁺

c. Synthesis of Compounds 2-4

Compound 2-3(1.2g,4.6mmol) and NCS (748.0mg,5.6mmol) were added to DMF (30 mL). The reaction solution was heated to 50 ℃ and the reaction was continued for 12 h. After completion of the reaction, the reaction solution was poured into water and extracted with ethyl acetate (30mL × 3). The organic phases were combined and dried over sodium sulfate. The organic phase was filtered and concentrated under reduced pressure and the crude product was purified by column chromatography (10-30% EA in PE) to give 0.708g of an off-white solid in 52.2% yield. MS Found 295.0[ M + H] ⁺ ,297.0[M+2+H] ⁺ .

d. Synthesis of Compounds 2-5

Compound 2-4(707.4mg,2.4mmol), EDCI (460.1mg,2.4mmol) and pyridine (379.7 mg,4.8mmol) were added to 10mL THF. The reaction was carried out at room temperature for 12 h. After the reaction was complete, water was added to quench and extracted with ethyl acetate (30mL x 3). The combined organic phases were washed with saturated brine (30mL) and dried over sodium sulfate. Filtering, and concentrating the organic phase under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (10-40% EA in PE) to give 0.5g of an off-white solid in 46.5% yield. MS Found 448.0[ M + H] ⁺ ,450.0[M +2+H] ⁺ .

e. Synthesis of Compound 2

To a solution of compound 2-5(493.1mg,1.1mmol) in DCM (10mL) was added trifluoroacetic acid (1 mL). The reaction is carried out for 12h at 30 ℃. The reaction was concentrated under reduced pressure and extracted with DCM (20mL × 3). The organic phases are combined and concentrated to give the crude product. The crude product was purified by prep-HPLC to give 94.1mg of a white solid in 24.6% yield. MS Found 348.1[ M + H] ⁺ ,350.1[M+2+H] ⁺ 。 ¹ H NMR(400MHz, DMSO-d ₆ ):δ6.38(s,1H),6.51-6.55(m,1H),6.91-6.99(m,2H),7.06-7.11(m,2H), 7.29-7.41(m,1H),7.42(dd,J＝4.8,2.0Hz,1H),8.58(m,1H),10.54(m,1H),11.17 (m,1H)。

Example 3: synthesis of Compound 3

To a mixed solution of 4-indolecarboxaldehyde (725.8mg,5.0mmol) and TFE/ACN (40mL, v/v ═ 1:1) was added 3-bromo-4-fluoroaniline (950.1mg,5.0 mmol). The reaction is carried out for 1h at room temperature. The reaction mixture was concentrated and dissolved in DCM/TFE mixture (20mL, v/v ═ 1:1) and TMSCN (595.3mg,6.0mmol) was added. The reaction is carried out for 3h at room temperature. The reaction was again concentrated and taken up in CHCl ₃ A solution of/THF (40mL, v/v ═ 1:1) was reconstituted. Manganese dioxide (4.3g,49.4mmol) was added and the reaction was heated to reflux and the reaction was continued for 2 days. After the reaction was completed, the mixture was filtered, and the filter cake was washed with methanol. The filtrate was collected and concentrated under reduced pressure to give a crude product. The crude product was purified by prep-HPLC to give 0.12g of a yellow solid in 7.0% yield. MS Found 342.1[ M + H] ⁺ ；344.1[M+2+H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ ):δ7.24(t,J＝2.0Hz,1H), 7.35-7.39(m,2H),7.55(t,J＝8.8Hz,1H),7.65(t,J＝2.8Hz,1H),7.72(dd,J＝6.0, 2.4Hz,1H),7.78-7.81(m,2H),11.75(s,1H)。

Example 4: synthesis of Compound 4

To a solution of 7-indolecarboxaldehyde (725.8mg,5.0mmol) in TFE/ACN (40mL, v/v ═ 1:1) was added 3-bromo-4-fluoroaniline (950.1mg,5.0 mmol). The reaction was stirred at room temperature for 1 h. The reaction was concentrated and the residue was dissolved with DCM/TFE (20mL, v/v ═ 1:1) and TMSCN (595.3mg,6.0mmol) was added. The reaction was continued at room temperature for 3 h. The reaction solution is concentrated again for the crude product _CHCl3 A solution of THF (40mL, v/v ═ 1:1) was redissolved, and manganese dioxide (4.3g,49.4mmol) was added. The reaction was heated to reflux and the reaction was continued for 2 days. After the reaction was completed, the mixture was filtered, and the filter cake was washed with methanol. The filtrate was collected and concentrated under reduced pressure to give a crude product. The crude product was purified by prep-HPLC to give 0.106g of a yellow solid in 6.2% yield. MS Found 342.1[ M + H] ⁺ ； 344.1[M+H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ ):δ6.68(dd,J＝3.2,2.0Hz,1H), 7.32(t,J＝7.6Hz,1H),7.48-7.52(m,2H),7.59(t,J＝8.8Hz,1H),7.80(d,J＝7.6 Hz,1H),7.85(dd,J＝6.4,2.4Hz,1H),7.98(d,J＝7.6Hz,1H),11.50(s,1H)。

EXAMPLE 5 Synthesis of Compound 5

a. Synthesis of Compound 5-2

To a solution of methyl 2-amino-3-nitrobenzoate (3.9g,19.9mmol) in isopropanol (30mL) was added iron powder (5.6g,100.3mmol), ammonium chloride (5.6g,104.7mmol) and formic acid (20 mL). The reaction solution was heated to 80 ℃ and the reaction was continued for 12 h. After the reaction is finished, filtering is carried out, and the filtrate is collected and concentrated to obtain a crude product. The crude product was purified by column chromatography (10-50% EAin PE) to afford 3.0g of an off-white solid with 85.6% yield. MS Found 177.0[ M + H] ⁺ 。

b. Synthesis of Compound 5-3

To a solution of compound 5-2(3.0g,17.0mmol) in THF (100mL) at room temperature was slowly added LiAlH ₄ (1.3g,34.3mmol) and stirred overnight. After the reaction was complete, water was added to quench and extracted with ethyl acetate (50mL × 3). The organic phase was collected and concentrated to give the crude product. The crude product was purified by column chromatography (20-60% EA in PE) to give 2.0g of an off-white solid in 79.4% yield. MS Found 149.2[ M + H ]] ⁺ 。

c. Synthesis of Compound 5-4

To a solution of compound 5-3(2.0g,13.5mmol) in DCM (50mL) was added manganese dioxide (11.7g,134.6mmol) at room temperature. The reaction was heated to reflux and the reaction was continued overnight. After the reaction is finished, filtering and reactingThe solution was taken, the filtrate was collected and concentrated to obtain crude product. The crude product was purified by column chromatography (20-60% EA in PE) to give 0.6g of an off-white solid in 30.4% yield. MS Found 147.0[ M + H ]] ⁺ 。

d. Synthesis of Compound 5

To a solution of compound 5-4(600.0mg,4.1mmol) in TFE/ACN (40mL, v/v ═ 1:1) was added 3-bromo-4-fluoroaniline (950.1mg,5.0mmol) at room temperature and stirring was continued for 1 h. The reaction was concentrated and the residue was taken up in DCM/TFE (20mL, v/v ═ 1:1) and TMSCN (595.3mg,6.0mmol) was added. The reaction was continued with stirring at room temperature for 3 h. The reaction was again concentrated and taken up in CHCl ₃ A solution of/THF (40mL, v/v ═ 1:1) was redissolved and manganese dioxide (4.3g,49.5mmol) was added. The reaction was heated to reflux and the reaction was continued for 2 days. After the reaction, the reaction solution was filtered, and the filter cake was washed with methanol. The filtrate was collected and concentrated under reduced pressure to give a crude product. The crude product was purified by prep-HPLC to give 0.132g of a yellow solid in 9.4% yield. MS Found 343.0[ M + H ]] ⁺ ；345.0[M+H+2] ⁺ 。 ¹ H NMR(400MHz,CDCl ₃ ):δ7.21-7.30(m, 2H),7.49-7.53(m,2H),8.03(d,J＝7.6Hz,1H),8.13(d,J＝8.0Hz,1H),8.22(s, 1H),11.00(brs,1H)。

Example 6: synthesis of Compound 6

a. Synthesis of Compound 6-2

Reacting 7-chloro-1H-pyrrolo [3,2-b ] at room temperature]Pyridine (2.7g,17.7mmol), Pd (dppf) Cl ₂ (658.5mg,0.9mmol) and Et ₃ N (2.8g,27.7mmol) was added to 20mL ethanol. And refluxing and reacting for 12h under the condition of a carbon monoxide balloon. Filtering after the reaction is finished, and concentrating the filtrate or obtaining a crude product. The crude product was purified by column chromatography (20-40% EA in PE) to give 1.6g of an off-white solid in 47.5% yield. MS Found 191.2[ M + H ]] ⁺ 。

b. Synthesis of Compound 6-3

To a solution of compound 6-2(1.6g,8.4mmol) in THF (30mL) at room temperature was slowly added LiAlH ₄ (0.9g,23.7mmol) and the reaction stirred overnight. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (20mL × 3), and the organic phases were combined and dried over sodium sulfate. Filtration and concentration of the organic phase gave 0.98g of crude grey solid in 78.7% yield which was directly used in the next reaction without further purification.

c. Synthesis of Compound 6-4

Manganese dioxide (4.6g,52.9mmol) was added to a solution of compound 6-4(977.9mg,6.6mmol) in DCM (30mL) at room temperature, heated to reflux and reacted overnight. After the reaction, the reaction solution was filtered and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (20-60% EAin PE) to give an off-white solid 0.7g, yield 72.6%. MS Found 147.3[ M + H ]] ⁺ 。

d. Synthesis of Compounds 6-5

To a solution of compound 6-4(686.9mg,4.7mmol) in TFE/ACN (40mL, v/v ═ 1:1) was added 3-bromo-4-fluoroaniline and a catalytic amount of acetic acid at room temperature, and the mixture was stirred overnight. After the reaction is finished, concentrating the reaction solution to obtain a crude product. The crude product was purified by column chromatography (20-60% EA in PE) to give 1.2g of an off-white solid in 80.3% yield.

e. Synthesis of Compound 6

To a solution of compound 6-5(318.1mg,1.0mmol) in ACN (20mL) were added TMSCN (496.1mg,5.0mmol), Zn (CN) ₂ (11.7mg,0.1mmol) and AlCl ₃ (13.3mg,0.1 mmol). The reaction was stirred at 50 ℃ for 4 h. Manganese dioxide (434.7mg,5.0mmol) was then added to the reaction solution and the reaction solution was heated to 80 ℃ overnight. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with ethyl acetate. The filtrate was collected and concentrated to give the crude product. The crude product was freshly purified layer by layer (50% of EA in PE) to give 0.25g of a yellow solid in 72.9% yield. MS Found 343.0[ M + H ]] ⁺ ,345.0[M+2+H] ⁺ .。 ¹ H NMR (400MHz,DMSO-d ₆ ):δ6.78-6.79(m,1H),7.58-7.65(m,3H),7.78(d,J＝3.2Hz, 1H),7.93(dd,J＝4.0,2.0Hz,1H),8.65(d,J＝5.2Hz,1H),11.60(brs,1H)。

Example 7: synthesis of Compound 7

a. Synthesis of Compound 7-2

At room temperature, to produce 7-bromo-1H-pyrrolo [3,2-c]To a solution of pyridine (1.8g,9.1mmol) in ethanol (20mL) was added Pd (dppf) Cl ₂ (658.5mg,0.9mmol) and triethylamine (2.8g,27.7 mmol). Refluxing reaction for 12h under the condition of carbon monoxide balloon. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (20-40% EA in PE) to give 1.4g of an off-white solid in 80.9% yield.

b. Synthesis of Compound 7-3

To a solution of compound 7-2(1.4g,7.4mmol) in THF (30mL) at room temperature was slowly added LiAlH ₄ (0.6g,15.8mmol) and the reaction stirred overnight. After the reaction was complete, water was added to quench and extracted with ethyl acetate (20mL × 3). The organic phase was collected and dried over sodium sulfate. Filtering and concentrating the organic phase to obtain a crude product. The crude product is purified by column chromatography (20-60% EA)in PE) to yield an off-white solid 0.8g, 73.0% yield. MS Found 149.2[ M + H ]] ⁺ 。

c. Synthesis of Compound 7-4

To a solution of compound 7-3(800mg,5.4mmol) in DCM (30mL) was added manganese dioxide (4.6g,52.9mmol) at room temperature and the reaction was refluxed overnight. After the reaction, the reaction solution was filtered and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (20-60% EAin PE) to give an off-white solid 0.6g, yield 76.0%. MS Found 147.3[ M + H ]] ⁺ 。

d. Synthesis of Compound 7-5

To a solution of compound 7-4(599.2mg,4.1mmol) in TFE/ACN (40mL, v/v ═ 1:1) at room temperature was added 3-bromo-4-fluoroaniline (779.0mg,4.1mmol) and a catalytic amount of acetic acid. The mixture was stirred overnight. After the reaction is finished, the crude product is obtained by concentration. The crude product was purified by column chromatography (20-60% EAin PE) to give an off-white solid 0.58g, yield 44.5%. MS Found 318.0[ M + H] ⁺ ,320.0[M+2+H] ⁺ 。

e. Synthesis of Compound 7

To compound 7-5(572.7mg,1.8mmol) in CHCl ₃ To a solution of/THF (40mL, v/v ═ 1:1) was added TMSCN (813.5mg,8.2mmol) and manganese dioxide (1.8g,20.7 mmol). Heating to 80 ℃ and reacting for 24 h. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with methanol. Collecting the filtrate, and concentrating to obtain a crude product. The crude product was purified by prep-HPLC to give 0.13g of a yellow solid in 21.4% yield. MS Found 343.0[ M + H ]] ⁺ ,345.0[M+2+H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ ):δ6.83-6.84(m,1H), 7.53-7.63(m,3H),7.87(dd,J＝3.6,2.4Hz,1H),8.77(s,1H),9.11(s,1H),11.84(brs, 1H)。

Example 8: test for inhibiting IDO1 Activity in Hela cells

The method comprises the following steps:

(1) reagents, materials, devices, apparatus

RPMI 1640 medium, phenol red (1X) free (invitrogen-11835-; 0.05% trypsin-EDTA (1X) (invitrogen-25300-; recombinant human interferon-gamma, trichloroacetic acid (TCA), 4-dimethylaminobenzaldehyde, glacial acetic acid; RPMI 1640 medium (1X)500ml final concentration 10%; penicillin/streptomycin (100 ×): final concentration of 100units/ml penicillin, 100. mu.g/ml streptomyces; spectramax 340pc (molecular device).

(2) Method and operation

The first day:

(1) the medium was discarded.

(2) The cell layer was washed with 10ml of PBS solution to remove trypsin inhibitor.

(3) 3ml of 0.05% pancreatin EDTA was added to the flask and incubated at 37 ℃ for 3-5 min.

(4) 10ml of complete growth medium was added to each flask and the cells were gently pipetted.

(5) The cell suspension was transferred to a 50ml conical tube.

(6) The cells were centrifuged at 1000rpm for 5min at room temperature.

(7) Cell concentrations were counted and cell viability was determined.

(8) Seed cells were seeded into 96-well plates per 200000 cells.

(10) 100 μ l IFN-. gamma.was added at a concentration of 100ng/ml to the whole plate.

(11) Compounds were added at different concentrations.

(12) Cells were incubated at 37 ℃ with 5% CO ₂ Culturing in an incubator for 20 h.

The next day:

(1) trichloroacetic acid at a concentration of 30% to a concentration of 0.2%. The cell plates were further incubated at 50 ℃ for 30 min.

(2) Centrifuge at 2400rpm for 10 min.

(3) Ehrlich reagent was added.

(4) Absorbance at 490nm was measured.

(5) Computing IC ₅₀ The value is obtained.

TABLE 1 inhibitory Activity of selected Compounds on IDO1 in Hela cells

In the table, "A" represents IC of the compound of example ₅₀ The value is less than 1. mu.M, "B" represents the IC of the compound of example ₅₀ The value is between 1. mu.M and 10. mu.M, "C" denotes the IC of the compound of the example ₅₀ The value is greater than 10. mu.M.

Claims

1. An imine compound represented by formula I or a pharmaceutically acceptable salt thereof:

wherein:

r1 is hydroxy, substituted or unsubstituted 5-8 membered aryl or heteroaryl;

r2 is cyano, substituted or unsubstituted 5-8 membered arylamino;

w is selected from C, N;

x is selected from C, N;

y is selected from C, N;

z is selected from C, N;

wherein W and X are not N at the same time, and Y and Z are not C at the same time.

2. The imine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein:

r1 is hydroxy, or substituted 5-8 membered aryl,

r2 is cyano, or a substituted 5-to 8-membered arylamino.

3. The imine compound or a pharmaceutically acceptable salt thereof according to claim 2, wherein:

r1 is hydroxy, or substituted phenyl,

r2 is cyano, or substituted phenylamino.

4. The imine compound according to claim 3, or a pharmaceutically acceptable salt thereof, wherein:

r1 is hydroxy, or phenyl substituted by halogen,

r2 is cyano, or phenylamino substituted with halogen.

5. The imine compound according to claim 4, or a pharmaceutically acceptable salt thereof, wherein:

r1 is hydroxy, or

R2 is cyano, or

6. The imine compound or a pharmaceutically acceptable salt thereof according to claim 5, selected from:

7. the imine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt is a salt of the imine compound with: hydrochloric acid, p-toluenesulfonic acid, tartaric acid, maleic acid, lactic acid, methanesulfonic acid, sulfuric acid, phosphoric acid, citric acid, acetic acid or trifluoroacetic acid.

8. The imine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein: the pharmaceutically acceptable salts further include solvates, metabolite prodrugs thereof.

9. A pharmaceutical composition comprising the imine compound according to claim 1 or a pharmaceutically acceptable salt thereof.

10. Use of the imine compound or the pharmaceutically acceptable salt thereof according to claim 1 or the pharmaceutical composition containing the imine compound or the pharmaceutically acceptable salt thereof according to claim 9 in the preparation of a medicament for treating tumors.