CN111518076A - Preparation method of indoleamine-2, 3-dioxygenase (IDO) inhibitor - Google Patents

Abstract

The present invention relates to a preparation method of the compound represented by formula I, and the compound prepared by the method of the present invention has the activity of inhibiting indoleamine 2,3-dioxygenase.

Description

Preparation method with indoleamine-2,3-dioxygenase (IDO) inhibitor

This application is a divisional application. The Chinese application number of the parent case is: 201980004340.2, the international application number is PCT/CN2019/071704, and the international filing date is January 15, 2019.

The present invention claims the priority of Chinese patent application CN201810044798.8, and the contents recorded in the description, drawings and claims of the priority document are fully incorporated into the description of the present invention and are taken as a part of the original description of the description of the present invention. The applicant further declares that the applicant has the right to amend the description and claims of the present invention based on this priority document.

technical field

The invention relates to the field of medicine, in particular to a preparation method with an indoleamine-2,3-dioxygenase (IDO) inhibitor.

Background technique

Tryptophan (TRP) is an alpha-amino acid used in protein biosynthesis. It contains an α-amino group, an α-carboxylic acid group and a side chain indole. It is essential to human beings, the human body cannot synthesize it, but must be obtained from the diet. Tryptophan is also a precursor for the synthesis of the neurotransmitter serotonin and the hormone N-acetyl-5-methoxytryptamine (melatonin). The heme-dependent enzyme indoleamine 2,3-dioxygenase (also called IDO, or IDO1) is the extrahepatic metabolic enzyme responsible for the conversion of tryptophan to N-formyl-kynurenine, which is tryptophan The first step in the process of acid metabolism is also the rate-limiting step in the entire process. N-formyl-kynurenine is the precursor of various biologically active molecules kynurenine (kynurenine, or Kyn), which has immunomodulatory functions (Schwarcz et al, Nat. Rev. Neurosci. 2012; 13(7):465).

Indoleamine 2,3-dioxygenase (IDO) is widely expressed in solid tumors (Uyttenhove et al, Nat. Med. 2003; 10:1269), both in primary and metastatic cancer cells. IDO is induced in tumors by proinflammatory factors, including type I and type II interferons produced by infiltrating lymphocytes (Tnani and Bayard, Biochim Biophys Acta. 1999; 1451(l):59; Mellor and Munn, Nat. Rev. Immunol. 2004; 4(10): 762; Munn, Front Biosci. 2012; 4: 734) and transforming growth factor-beta (TGF-beta) (Pallotta et al, Nat. Immunol. 2011; 12(9):870 ). In recent years, increasing evidence has shown that IDO, as an inducible enzyme, plays an important role in immune cell regulation. Decreased tryptophan levels and increased kynurenine suppress immune effector cells and promote adaptive immunosuppression by inducing and maintaining regulatory T cells; the concentration of tryptophan in the immune system is positively correlated with T cells. In the tumor immune microenvironment, activated or overexpressed IDO leads to tryptophan depletion, which leads to T cell death, immune system inactivation, and ultimately tumor immune tolerance and immune escape. Existing studies have shown that the imbalance of immune balance caused by IDO is deeply involved in the generation and progression of tumors. Therefore, IDO has become an important target for immunotherapy such as tumors. In addition to being associated with tumors, IDO is also associated with viral infections, depression, organ transplant rejection or autoimmune diseases (Johnson and Munn, Immunol. Invest. 2012; 41(6-7):765). Therefore, drugs targeting IDO are also of great value for the treatment of the above-mentioned diseases. In conclusion, the development of active and selective IDO inhibitors can effectively treat diseases caused by harmful substances in the kynurenine pathway by modulating the kynurenine channel and maintaining tryptophan levels in the body, whether as a Either single-agent or combination therapy is necessary.

Numerous published preclinical data also further confirm the role of IDO in the antitumor immune response. IDO inhibitors can be used to activate T cells, thereby enhancing T cell activation when T cells are suppressed by pregnancy, malignancy, or viruses such as HIV. Forced IDO induction in cancer cells has been shown to have a survival advantage (Uyttenhove et al, Nat Med. 2003; 10:1269). Another in vivo study showed that IDO inhibitors reduce the reliance on lymphocytes in tumor growth by reducing kynurenine levels (Liu et al, Blood. 2010; 115(17):3520). Preclinical studies have also shown that IDO inhibitors have synergistic effects if used in combination with other oncology drugs, such as radiotherapy, chemotherapy or vaccines (Koblish et al, Mol. Cancer Ther. 2010; 9(2): 489; Hou et al, Cancer Res. 2007;67(2):792; Sharma et al, Blood. 2009;113(24):6102).

The research of IDO inhibitor antitumor drugs has made important progress in the world, such as INCB024360, NLG919 and BMS-986205 have entered the clinic. However, due to the problem of toxic and side effects of INCB024360, the current clinical research dose (50mg bid, or 100mg bid) is about 30% of the optimal dose (300mg bid, 600mg bid), and the clinical activity is greatly limited. At the same time, the toxicity base of INCB024360 The group is also a pharmacophore, and INCB024360 and its derivatives have the problem of high toxicity. The safety of NLG919 is good, but the biological activity of NLG919 is poor. BMS-986205 has also entered the clinic, but clinical data are limited. There is also a need for better IDO inhibitors for tumor therapy.

SUMMARY OF THE INVENTION

One aspect of the present invention provides a compound represented by formula (I), its salt, solvate, prodrug, metabolite, nitrogen oxide, stereoisomer or isotopic derivative:

表示：-、

或者

Cy₁选自任选地被取代基取代的5-15元环，所述的取代基选自：卤素、羟基、C_1-6烷基、氨基、卤代C_1-6烷基、巯基、C_1-6烷基巯基、C_1-6烷基氨基、二(C_1-6烷基)氨基和氰基；in

express:-,

or

Cy ₁ is selected from a 5-15-membered ring optionally substituted by a substituent selected from the group consisting of: halogen, hydroxyl, C _1-6 alkyl, amino, halogenated C _1-6 alkyl, mercapto, C _1-6 alkylmercapto, C _1-6 alkylamino, di(C _1-6 alkyl)amino and cyano;

Cy ₂ is selected from C _6-10 membered cycloalkyl, C _6-10 membered heterocyclyl, C _6-10 membered aryl or C _6-10 optionally substituted by one, two or more R ² Member heteroaryl; preferably phenyl, pyridyl, cyclohexyl, piperidinyl, piperazinyl, pyrazinyl, pyrimidinyl, morpholinyl substituted with one, ^two or more R2; pyridazine base;

R ¹ and R ² are independently selected from hydrogen atom, halogen, hydroxyl, nitro, cyano, sulfonic acid, C _1-6 alkyl, C _3-6 cycloalkyl, C _2-6 alkenyl, C _{2 -6alkynyl} , C _1-6 alkoxy, halogenated C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkoxy, halogenated C ₃ -C ₆ cycloalkyl, C _1-6 alkane sulfanyl, C _1-6 alkylcarbonyl, C _1-6 alkoxycarbonyl, di(C _1-6 alkyl) amino, C _2-6 alkoxycarbonyl, amino, C _1-6 alkylamino, Di(C _1-6 alkyl)amino, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl) carbamoyl, di(C _1-6 alkyl) amino C _{2 -6} alkylcarbamoyl, sulfamoyl, C _1-6 alkyl sulfamoyl, di(C _1-6 alkyl) sulfamoyl, di (C _1-6 alkyl) amino C _2-6 alkane Sulfamoyl, C _1-6 alkylsulfonyl, C _1-6 alkylsulfinyl, di(C _1-6 alkyl) phosphono, hydroxy C _1-6 alkyl, hydroxycarbonyl C _1-6 Alkyl, C _1-6 alkoxy C _1-6 alkyl, C _1-6 alkylsulfonyl C _1-6 alkyl, C _1-6 alkylsulfinyl C _1-6 alkyl, di( C _1-6 alkyl) phosphono C _1-6 alkyl, hydroxy C _2-6 alkoxy, C _1-6 alkoxy C _2-6 alkoxy, amino C _1-6 alkyl, C _{1 -6} alkylamino C _1-6 alkyl, di(C _1-6 alkyl) amino C _1-6 alkyl, di (C _1-6 alkyl) aminoacetyl, amino C _2-6 alkoxy , C _1-6 alkylamino C _2-6 alkoxy, two (C _1-6 alkyl) amino C _2-6 alkoxy, hydroxy C _2-6 alkyl amino, C _1-6 alkoxy C _2-6 alkylamino, amino C _2-6 alkyl amino, C _1-6 alkyl amino C _2-6 alkyl amino, di(C _1-6 alkyl) amino C _2-6 alkyl amino; Or two adjacent R ¹ or R ² are cyclized with each other to form a 3-8 membered ring, and the ring contains 0-3 heteroatoms;

m, n are integers selected from 0, 1, 2, 3 and 4;

R ^a and R ^b are each independently selected from hydrogen, C ₁ -C ₆ alkyl or C _3-6 cycloalkyl;

X is selected from CR ^a R ^b , NR ^e or O;

Y is selected from CR ^e or N; wherein R ^e represents hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl, C _1-6 haloalkyl.

Another aspect of the present invention provides a compound represented by formula (II), its salt, solvate, prodrug, metabolite, nitrogen oxide, stereoisomer or isotopic derivative:

Wherein, W ₁ , W ₂ , W ₃ and W ₄ are independently selected from CR ^e , C=O or N; p is an integer selected from 0, 1, 2, 3 and 4; R ¹ , R ² , Cy ¹ , R ^a , R ^b , ^Re , X, Y, m, n are as defined in formula I; dashed lines represent single or double bonds.

In one embodiment of the present invention, Cy ₁ is selected from the following groups:

wherein R ₃ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl;

The above-mentioned groups may be replaced by one or more than two selected from halogen, hydroxyl, C _1-6 alkyl, amino, halogenated C _1-6 alkyl, mercapto, C _1-6 alkyl mercapto, C _1-6 alkane substituted with substituents of amino, di(C _1-6 alkyl) amino and cyano.

One aspect of the present invention provides a compound represented by formula (III), its salt, solvate, prodrug, metabolite, nitrogen oxide, stereoisomer or isotopic derivative:

wherein R ¹ , R ² , R ^a , R ^b , X, Y, W ₁ , W ₂ , W ₃ , W ₄ , m, n, p are as defined in formula II.

One aspect of the present invention provides a compound represented by formula (IV), its salt, solvate, prodrug, metabolite, nitrogen oxide, stereoisomer or isotopic derivative:

wherein Q ₁ and Q ₂ are independently selected from CR ^a R ^b , NR ^e or O; Q ₃ is selected from CR ^a or N; wherein R ¹ , R ² , R ^a , R ^b , ^Re , X, Y , W ¹ , W ² , W ₃ , W ₄ , m, p are as defined in formula II;

In another embodiment of the present invention, the compound of formula II has the structure of formula (V):

wherein R ¹ , R ² , R ^a , R ^b , X, Y, W ₁ , W ₂ , W ₃ , W ₄ , m, p are as defined in Formula II above.

表示-、

或者

在本公开中，

优选为

In this disclosure,

express-,

or

In this disclosure,

preferably

Unless otherwise specified, all compound structures of the present invention also include possible stereoisomers (including: enantiomeric, diastereomeric, geometric, conformational, androtational). For example, the R and S configurations of each chiral center, and the E and Z isomers of each olefinic double bond are included in this invention. For some freely rotating bonds, the positions of the substituents can also be freely rotated, for example:

也同时代表：

也代表其互变异构体：

所以，一个单一的立体化学异构体，以及其对映体混合物，几何异构体混合物，构象异构体混合物，互变异构体都在本专利申请范畴。Structural formula

Also represents:

Also represents its tautomers:

Therefore, a single stereochemical isomer, as well as its enantiomeric mixtures, geometric isomer mixtures, conformational isomer mixtures, and tautomers are all within the scope of this patent application.

The compounds of the present invention can also be prepared in the form of pharmaceutically acceptable salts formed using, for example, the following inorganic or organic acids: hydrochloric, hydrobromic, sulfuric, phosphoric, nitric, acetic, glycolic, Lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzene Formic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid and toluenesulfonic acid. When referring to the compounds of the present invention, these pharmaceutically acceptable salts of the compounds of the present invention are also encompassed.

The pharmaceutically acceptable salts of the present invention can be prepared by conventional methods, for example, by dissolving a compound of the present invention in a water-miscible organic solvent (eg, acetone, methanol, ethanol, and acetonitrile), adding thereto an excess of an organic acid or inorganic An aqueous acid solution to precipitate the salt from the resulting mixture, the solvent and remaining free acid removed therefrom, and the precipitated salt isolated.

Design and React Examples

AD-H手性柱(0.46cm I.D.×15cm L,HEP:ETOH(0.1％DEA)＝60:40(V/V))分离。The compounds of the present invention can be synthesized by known procedures with reference to the following description. All purchased solvents and reagents were used without treatment. All synthesized compounds can be analyzed and verified by, but not limited to, the following methods: LCMS (liquid chromatography mass spectrometry, liquid mass spectrometry) and NMR (nuclear magneticresonance, nuclear magnetic resonance). Nuclear magnetic resonance (NMR) was measured by Bruker AVANCE-500 nuclear magnetic instrument, and the deuterated solvents used in the measurement were deuterated dimethyl sulfoxide (d6-DMSO), deuterated chloroform (CDCl3), and tetramethylsilane (TMS) as internal standard. The following abbreviations represent various types of split peaks: singlet (s), doublet (d), triplet (t), multiplet (m), broad (br). Mass spectrometry (MS) was measured by Thermo Fisher-MSQ Plus LC/MS, and chiral compounds were resolved by Daicel

AD-H chiral column (0.46 cm ID x 15 cm L, HEP:ETOH (0.1% DEA) = 60:40 (V/V)) separation.

The compounds of the present invention can be prepared by referring to the following.

General route one:

Synthesis of Intermediate F (cis and trans mixture, racemic compound)

The first step: ethyl 4-oxocyclohexane (2.0 g, 10.86 mmol) was dissolved in 60 mL of ultra-dry tetrahydrofuran, and bis(trimethylsilicon was added dropwise to the solution under nitrogen atmosphere at -78 °C base) sodium amide (2 mol/L tetrahydrofuran solution) (6.5 mL, 13.03 mmol). The reaction solution was stirred at this temperature for 1 hour. Then a solution of N-phenylbis(trifluoromethanesulfonyl)imide (4.65 g, 13.03 mmol) in tetrahydrofuran (20 mL) was added. After the dropwise addition, the reaction mixture was continued to stir at room temperature overnight until the reaction material was completely consumed as detected by TLC. The reaction solution was quenched with 5 mL of aqueous potassium hydrogen sulfate solution, the solid was removed by filtration, and the filtrate was concentrated. 50 mL of methyl tert-butyl ether was added to the residue, and the organic layer was washed with 1.0 mol/L sodium hydroxide solution (3×20 mL) and 20 mL of saturated brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain Intermediate A (3.12 g) as an orange-red oily liquid with a yield of 91%. ¹ H NMR (500 MHz, CDCl ₃ ) δ 5.74-5.70 (m, 1H), 4.15 (q, J=7.0 Hz, 2H), 2.48-2.40 (m, 1H), 2.38-2.32 (m, 2H), 2.30(d,J=7.0Hz,2H),2.18-2.10(m,1H),1.97-1.89(m,2H),1.57-1.48(m,1H),1.27(t,J=7.0Hz,3H) .

The second step: Intermediate A (3.12g, 9.86mmol) was dissolved in 15mL of dioxane, followed by adding pinacol biboronate (3.26g, 12.82mmol), potassium acetate (2.90g, 29.59mmol), Sodium bromide (406 mg, 3.95 mmol) and Pd(dppf)Cl2 (722 _mg , 0.98 mmol). The reaction mixture was refluxed under nitrogen atmosphere overnight. Then dioxane, the reaction solvent, was evaporated to dryness, ethyl acetate was added, filtered through celite, the filtrate was concentrated and then separated by a rapid column machine to obtain Intermediate B (1.66 g) as a colorless liquid with a yield of 57%. ¹ H NMR (500MHz, CDCl ₃ ) δ 6.54-6.48 (m, 1H), 4.12 (q, J=6.5Hz, 2H), 2.30-2.02 (m, 7H), 1.84-1.72 (m, 2H), 1.27–1.23 (m, 15H).

The third step: Intermediate B (1.66g, 5.64mmol) was dissolved in 12mL/3mL dioxane/water, 4-chloro-6-fluoroquinoline (860mg, 4.74mmol), potassium carbonate (1.96g) were added successively , 14.21 mmol) and Pd( _PPh3 ) ₄ (274 mg, 0.24 mmol). The reaction mixture was refluxed under nitrogen atmosphere overnight. The reaction solution was then concentrated, diluted with 50 mL of water, extracted with ethyl acetate (3×50 mL), and the organic phase was concentrated and separated with a rapid column machine to obtain Intermediate C (1.48 g) as a pale yellow liquid with a yield of 100%. MS (ESI): m/z 313.9 (M+H) ⁺ . ¹ H NMR (500 MHz, CDCl ₃ ) δ 8.81 (d, J=4.5 Hz, 1 H), 8.16 (dd, J=8.5, 5.5 Hz) ,1H),7.62(dd,J＝10.0,2.5Hz,1H),7.52-7.46(m,1H),7.22(d,J＝4.5Hz,1H),5.86-5.81(m,1H),4.19( q, J=7.0Hz, 2H), 2.56–2.26 (m, 6H), 2.08–1.98 (m, 2H), 1.64–1.55 (m, 1H), 1.30 (t, J=7.0Hz, 3H).

The fourth step: Intermediate C (1.48 g, 4.72 mmol) was dissolved in 30 mL of ethanol, and 10% palladium on carbon (300 mg) was added. The reaction mixture was stirred at room temperature under a hydrogen atmosphere overnight. The palladium carbon was then filtered off through Celite, and the filtrate was concentrated. The residue was separated by a rapid column machine to obtain Intermediate D (1.31 g) as a pale yellow liquid with a yield of 88%. MS (ESI): m/z 316.0 (M+H) ⁺ . ¹ H NMR (500 MHz, CDCl ₃ ) δ 8.84–8.79 (m, 1H), 8.13 (dd, J=9.0, 5.5 Hz, 1H) , 7.66 (dd, J=10.5, 2.5Hz, 1H), 7.51–7.44 (m, 1H), 7.34 (d, J=4.5Hz, 1H), 4.20–4.14 (m, 2H), 3.26–3.18 (m , 1H), 2.53–2.43 (m, 2H), 2.31 (d, J=7.0Hz, 1H), 2.07–1.97 (m, 2H), 1.90–1.70 (m, 5H), 1.68–1.58 (m, 1H) ),1.31–1.25(m,3H).

Fifth step: Diisopropylamine (1.54 g, 15.22 mmol) was dissolved in 18 mL of tetrahydrofuran. To this solution was added dropwise a 2.5M solution of n-butyllithium (6.1 mL, 15.22 mmol) in n-hexane under nitrogen atmosphere at -78°C. A solution of Intermediate D (2.4 g, 7.61 mmol) in tetrahydrofuran (6 mL) was then added dropwise. The reaction mixture was stirred for an additional 1.5 hours at -78°C. Then methyl iodide (2.16 g, 15.22 mmol) was added dropwise, and the reaction mixture was warmed to room temperature and stirred overnight. The reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate (3×50 mL), the organic phases were combined, washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by a rapid column machine to obtain Intermediate E (1.96 g) as a pale yellow liquid with a yield of 78%. MS (ESI): m/z 330.5 (M+H) ⁺ . ¹ H NMR (500 MHz, CDCl ₃ ) δ 8.84–8.79 (m, 1H), 8.16–8.10 (dd, 1H), 7.66 (d, J=10.5Hz, 1H), 7.51–7.44 (m, 1H), 7.35 (d, J=4.5Hz, 1H), 4.22–4.14 (m, 2H), 3.32–3.23 (m, 1H), 2.82–2.72 (m, 1H), 2.12–1.98 (m, 2H), 1.96–1.55 (m, 7H), 1.32–1.24 (m, 3H), 1.20 (d, J=6.5Hz, 3H).

The sixth step: Intermediate E (400 mg, 1.21 mmol) was dissolved in 4 mL/4 mL of tetrahydrofuran/ethanol, and 2 mL of water was added. Sodium hydroxide (243 mg, 6.07 mmol) was then added to the solution. The reaction mixture was stirred at 50°C overnight and concentrated. Add 3 mL of water to dilute, adjust pH=3 with 4 mol/L hydrochloric acid solution, and filter to obtain Intermediate F (330 mg) as a white solid with a yield of 90%. MS (ESI): m/z 302.6 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.22 (s, 1H), 8.81 (d, J=4.5Hz, 1H), 8.14 –8.06(m,1H),8.01-7.94(m,1H),7.66(t,J=8.5Hz,1H),7.52(s,1H),3.32-3.23(m,1H),2.76-2.66(m , 1H), 1.97–1.62 (m, 7H), 1.61–1.51 (m, 1H), 1.49–1.31 (m, 1H), 1.09 (d, J=6.5Hz, 3H).

General route 2: asymmetric synthesis route

The asymmetric synthesis method of the intermediate K' adopts the synthesis method reported in the literature (WO2016073774A2)

General route three:

The first step: Intermediate F (or K', 1.0eq) was dissolved in N,N-dimethylformamide, and HATU (1.1eq) and diisopropylethylamine (3.0eq) were added. Substituted 1,2-diamine or substituted o-aminoaniline (1.5eq) was added to the reaction solution. The reaction mixture was stirred at 30°C overnight. Then, water and ethyl acetate were added to the reaction solution, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude intermediate that was used in the next step without purification.

The second step: The crude intermediate (1.0 eq) obtained in the previous step was dissolved in acetic acid, the mixture was stirred and reacted at 100° C. for 19 hours, and then the reaction solution was concentrated. The residue was purified by reverse phase high performance liquid chromatography to give the final compound.

Common route four:

The first step: Triethyl phosphonoacetate (968 mg, 4.32 mmol) was dissolved in 16 mL of ultra-dry tetrahydrofuran, and sodium tert-butoxide (415 mg, 4.32 mmol) was added under an ice bath at 0°C. After 10 minutes, a solution of Intermediate E' (1 g, 4.12 mmol) in tetrahydrofuran (4 mL) was added to the reaction. After 2 hours of reaction, it was quenched with water. The aqueous solution was extracted three times with 20 mL of ethyl acetate, and the organic phases were combined, washed with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by a flash column machine to give Intermediate F" (1.18 g) as a white solid in 92% yield. MS (ESI): m/z 314.0 (M+H) ^{+ .1H} NMR (500MHz, CDCl) ₃ ) δ8.81 (d, J=4.5Hz, 1H), 8.17 (dd, J=9.0, 5.5Hz, 1H), 7.72 (dd, J=10.0, 2.5Hz, 1H), 7.53–7.47 (m, 1H), 7.28 (d, J=4.5Hz, 1H), 5.75 (s, 1H), 4.19 (q, J=7.0Hz, 2H), 3.52–3.42 (m, 1H), 2.54–2.48 (m, 2H) ), 2.26–2.11 (m, 4H), 1.80–1.68 (m, 2H), 1.30 (t, J=7.0Hz, 3H).

The second step: NaH (383 mg, 9.57 mmol) was added to 15 mL of dimethyl sulfoxide, and trimethyl sulfoxide (2.11 g, 9.57 mmol) was added to the suspension. The mixture was stirred at room temperature for 1.5 hours. A solution of Intermediate F" (1.0 g, 3.19 mmol) in dimethyl sulfoxide (5 mL) was then added to the reaction. The reaction was stirred at room temperature overnight. It was then quenched with water, extracted with The intermediate G" (820 mg) was obtained by column separation as a colorless oily liquid with a yield of 78%. MS (ESI): m/z 328.1 (M+H) ⁺ . ¹ H NMR (500 MHz, CDCl ₃ ) δ 8.83 (d, J=4.5 Hz, 1 H), 8.24 (dd, J=9.0, 5.5 Hz) ,1H),7.71(dd,J＝10.0,2.5Hz,1H),7.55–7.49(m,1H),7.35(d,J＝4.5Hz,1H),4.19(q,J＝7.0Hz,2H) , 3.32–3.24 (m, 1H), 2.17 (td, J=13.0, 3.5Hz, 1H), 2.07–1.90 (m, 4H), 1.87–1.78 (m, 1H), 1.58 (dd, J=8.0, 5.5Hz, 1H), 1.46–1.37 (m, 1H), 1.30 (t, J=7.0Hz, 3H), 1.28–1.24 (m, 2H), 1.16–1.11 (m, 1H), 1.00 (dd, J =8.0,4.5Hz,1H).

The 3rd step: the intermediate G " (200mg, 0.61mmol) is dissolved in 10mL of ethanol, and 4mL of 2mol/L sodium hydroxide solution is added. The reaction solution is heated to 50 ° C and reacted for 2 hours. The /L hydrochloric acid solution was neutralized to pH=1. The aqueous phase was extracted with ethyl acetate. The organic phases were combined, dried with anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by preparative thin layer chromatography to obtain intermediate H" (150 mg ), white solid, 83% yield. MS (ESI): m/z 300.0 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.02 (br, 1H), 8.83 (d, J=4.5Hz, 1H), 8.10 (dd, J=9.0, 5.5Hz, 1H), 8.03 (dd, J=10.0, 2.5Hz, 1H), 7.71–7.64 (m, 1H), 7.38 (d, J=4.5Hz, 1H), 3.48– 3.41 (m, 1H), 2.21–2.13 (m, 1H), 2.01–1.80 (m, 4H), 1.75–1.65 (m, 1H), 1.51 (dd, J=8.0, 5.5Hz, 1H), 1.38– 1.32 (m, 1H), 1.11–1.05 (m, 1H), 1.04–0.99 (m, 1H), 0.95 (dd, J=7.5, 4.0Hz, 1H).

Step 4: Dissolve intermediate H" (1.0eq) in N,N-dimethylformamide, add HATU (1.1eq) and diisopropylethylamine (3.0eq). Then add to the reaction solution Substituted 1,2-diamine or substituted o-aminoaniline (1.5eq). The reaction mixture was stirred at 30°C overnight. Water and ethyl acetate were then added to the reaction solution, the organic phase was washed with saturated brine, and anhydrous sulfuric acid Dry over sodium, filter and concentrate. The obtained crude intermediate is dissolved in acetic acid, the mixture is stirred at 100° C. for 19 hours, and then the reaction solution is concentrated. The residue is purified by reverse-phase high performance liquid chromatography to obtain the final compound.

General route five:

The first step: n-butyllithium (0.49 mL, 1.22 mmol) was added dropwise to a solution of diisopropylamine (123 mg, 1.22 mmol) in tetrahydrofuran (15 mL) at -78°C. To this was added dropwise a solution of Intermediate G" (200 mg, 0.61 mmol) in tetrahydrofuran (5 mL). The reaction was stirred at -78°C for 1 hour. To the reaction was then added dropwise methyl iodide (173 mg, 1.22 mmol) in tetrahydrofuran ( 2mL) solution, the reaction was maintained at -78°C for half an hour and then warmed to room temperature and stirred overnight. Quenched with saturated ammonium chloride solution, the aqueous phase was extracted with ethyl acetate. The organic phases were combined, dried with anhydrous sodium sulfate, filtered, Concentrated. The residue was isolated by preparative thin layer chromatography to give compound intermediate H''' (121 mg) as a colorless oily liquid in 58% yield. MS(ESI): m/z342.4(M+H) ⁺ .

The second step: the intermediate H"' (100mg, 0.29mmol) was dissolved in 10mL of ethanol, and 2mL of 2mol/L sodium hydroxide solution was added. The reaction solution was heated to 50 ° C and reacted for 2 hours. 4mol/L hydrochloric acid solution was neutralized to pH=1. The aqueous phase was extracted with ethyl acetate. The organic phases were combined, dried with anhydrous sodium sulfate, filtered, and concentrated. The residue was separated by preparative thin layer chromatography to obtain intermediate I"' (76 mg), white solid, 83% yield. MS(ESI): m/z314.3(M+H) ⁺ .

The third step: Dissolve intermediate I"' (1.0eq) in N,N-dimethylformamide, add HATU (1.1eq) and diisopropylethylamine (3.0eq). Then add to the reaction solution Substituted 1,2-diamine or substituted o-aminoaniline (1.5eq) was added. The reaction mixture was stirred at 30°C overnight. Then, water and ethyl acetate were added to the reaction solution, and the organic phase was washed with saturated brine, anhydrous Dry over sodium sulfate, filter, and concentrate. The obtained crude intermediate was dissolved in acetic acid, and the mixture was stirred at 100° C. for 19 hours, and then the reaction solution was concentrated. The residue was purified by reverse-phase high-performance liquid chromatography to obtain the final compound.

Example 1: Compound 1

Compound 1 was prepared via general route one and general route three starting from intermediate F (20 mg) and 4-chloro-1,2-phenylenediamine. Compound 1 (10.05 mg) was obtained as a white solid with a yield of 37%. MS (ESI): m/z 408.3 (M+H) ⁺ . ¹ HNMR (500MHz, d ₆ -DMSO) δ 12.40 (s, 1H), 8.79 (d, J=4.5Hz, 1H), 8.11– 8.04(m, 1H), 7.97(d, J=10.5Hz, 1H), 7.65(t, J=8.5Hz, 1H), 7.60–7.45(m, 2H), 7.42(d, J=4.0Hz, 1H) ), 7.15 (d, J=8.5Hz, 1H), 3.30–3.24 (m, 1H), 2.95–2.88 (m, 1H), 1.95 (t, J=10.5Hz, 2H), 1.90–1.79 (m, 2H), 1.61–1.47 (m, 3H), 1.45–1.32 (m, 5H).

Example 2: Compound 17 and Compound 18

Compound 17 (trans, racemic) and compound 18 (cis, racemic) proceeded from intermediate F (40 mg) and 4-fluoro-1,2-phenylenediamine via general route one and general route three prepared.

Compound 17 (8.49 mg) was obtained, the first eluting isomer, as a white solid in 16% yield. MS(ESI): m/z 392.5(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.28(s, 1H), 8.79(s, 1H), 8.11–8.04(m, 1H), 7.97(d, J＝10.5Hz, 1H), 7.65(t, J＝8.5Hz, 1H), 7.45-7.39(m, 2H), 7.34(d, J＝10.0Hz, 1H), 7.02- 6.92 (m, 1H), 3.29–3.23 (m, 1H), 2.93–2.87 (m, 1H), 2.00–1.91 (m, 2H), 1.89–1.79 (m, 2H), 1.61–1.46 (m, 3H) ),1.45–1.33(m,5H).

Compound 18 (9.24 mg) was obtained, the second eluting isomer, as a white solid in 18% yield. MS(ESI): m/z 392.5(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.31(s, 1H), 8.86(s, 1H), 8.13–8.06(m, 1H), 7.97(d, J=11.0Hz, 1H), 7.66(t, J=8.5Hz, 1H), 7.58(s, 1H), 7.44–7.39(m, 1H), 7.23(d, J=9.0 Hz, 1H), 7.01–6.92 (m, 1H), 3.45–3.35 (m, 2H), 2.15–2.09 (m, 1H), 2.06–1.99 (m, 1H), 1.93–1.83 (m, 2H), 1.82–1.71 (m, 2H), 1.68–1.60 (m, 1H), 1.59–1.53 (m, 1H), 1.34 (d, J=6.5Hz, 3H), 1.21–1.14 (m, 1H).

Example 3: Compound 21 and Compound 22

Compound 21 (trans, rac) and compound 22 (cis, rac) from intermediate F (40 mg) and 4-methyl-1,2-phenylenediamine via general route one and general route Three are prepared.

Compound 21 (11.40 mg) was obtained, the first eluting isomer, as a white solid in 22% yield. MS(ESI): m/z 388.5(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 11.98(s, 1H), 8.79(s, 1H), 8.11–8.04(m, 1H), 7.97(d, J＝10.5Hz, 1H), 7.65(t, J＝8.5Hz, 1H), 7.44-7.38(m, 2H), 7.20(s, 1H), 6.93(t, J＝9.5 Hz, 1H), 3.30–3.23 (m, 1H), 2.91–2.83 (m, 1H), 2.39 (s, 3H), 2.00–1.91 (m, 2H), 1.89–1.78 (m, 2H), 1.61– 1.46(m,3H),1.44–1.33(m,5H).

Compound 22 (13.51 mg) was obtained, the second eluting isomer, as a white solid in 26% yield. MS(ESI): m/z 388.5(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.02(s, 1H), 8.86(s, 1H), 8.13–8.06(m, 1H), 7.97(d, J＝11.0Hz, 1H), 7.66(t, J＝9.0Hz, 1H), 7.58(s, 1H), 7.39(d, J＝8.5Hz, 1H), 7.20(s, 1H), 6.93(t, J＝10.0Hz, 1H), 3.46-3.34(m, 2H), 2.38(s, 3H), 2.16-2.09(m, 1H), 2.06-1.99(m, 1H), 1.93 –1.83 (m, 2H), 1.82–1.70 (m, 2H), 1.67–1.51 (m, 2H), 1.33 (d, J=6.5Hz, 3H), 1.21–1.14 (m, 1H).

Example 4: Compound 13 and Compound 14

Compound 13 (trans, racemic) and compound 14 (cis, racemic) proceeded from intermediate F (50 mg) and 4-chloro-1,2-phenylenediamine via general route one and general route three prepared.

Compound 13 (7.93 mg) was obtained, the first eluting isomer, as a white solid in 12% yield. MS(ESI): m/z 408.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.36(s, 1H), 8.80(s, 1H), 8.11–8.04(m, 1H), 7.97(d, J=11.0Hz, 1H), 7.65(t, J=8.5Hz, 1H), 7.60(s, 1H), 7.46–7.40(m, 2H), 7.15(t, J=10.5 Hz, 1H), 3.30–3.23 (m, 1H), 2.95–2.88 (m, 1H), 2.00–1.91 (m, 2H), 1.89–1.79 (m, 2H), 1.61–1.46 (m, 3H), 1.45–1.32(m,5H).

Compound 14 (10.25 mg) was obtained, the second eluting isomer, as a white solid in 15% yield. MS (ESI): m/z 408.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.42 (s, 1H), 8.86 (d, J=4.0 Hz, 1H), 8.13 –8.07(m,1H),7.98(d,J=11.0Hz,1H),7.67(t,J=8.5Hz,1H),7.61–7.56(m,2H),7.45(d,J=8.5Hz, 1H), 7.17–7.11 (m, 1H), 3.45–3.37 (m, 2H), 2.16–2.09 (m, 1H), 2.07–2.00 (m, 1H), 1.93–1.83 (m, 2H), 1.82– 1.71 (m, 2H), 1.68–1.60 (m, 1H), 1.59–1.52 (m, 1H), 1.34 (d, J=6.5Hz, 3H), 1.19–1.12 (m, 1H).

Example 5: Compound 33 and Compound 34

Compound 33 (trans, rac) and compound 34 (cis, rac) were prepared from intermediate F (40 mg) and 4-trifluoromethyl-1,2-phenylenediamine via general routes one and Prepared by general route three.

Compound 33 (5.95 mg) was obtained, the first eluting isomer, as a white solid in 10% yield. MS (ESI): m/z 442.4 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.63 (s, 1H), 8.79 (s, 1H), 8.10–8.04 (m, 1H), 7.97(d, J＝11.0Hz, 1H), 7.91(s, 1H), 7.67-7.61(m, 2H), 7.49-7.40(m, 2H), 3.30-3.23(m, 1H), 3.02 –2.94(m,1H),2.00–1.91(m,2H),1.91–1.82(m,2H),1.61–1.47(m,3H),1.46–1.35(m,5H).

Compound 34 (4.55 mg) was obtained, the second eluting isomer, as a white solid in 8% yield. MS(ESI): m/z 442.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.66(s, 1H), 8.87(s, 1H), 8.12–8.06(m, 1H), 7.98(d, J＝10.5Hz, 1H), 7.90(s, 1H), 7.69-7.62(m, 2H), 7.59(s, 1H), 7.48-7.41(m, 1H), 3.52-3.38 (m, 2H), 2.21–2.11 (m, 1H), 2.09–2.01 (m, 1H), 1.94–1.73 (m, 4H), 1.69–1.61 (m, 1H), 1.60–1.54 (m, 1H) ,1.37(d,J＝6.0Hz,3H),1.19–1.13(m,1H).

Example 6: Compound 27 and Compound 28

Compound 27 (trans, rac) and compound 28 (cis, rac) from intermediate F (50 mg) and 4-methoxy-1,2-phenylenediamine via general route one and general Route three is prepared.

Compound 27 (3.67 mg) was obtained, the first eluting isomer, as a white solid in 5% yield. (ESI): m/z 404.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 11.99(s, 1H), 8.80(s, 1H), 8.10–8.07(m, 1H ),7.98(d,J＝10.0Hz,1H),7.69-7.62(m,1H),7.46-7.39(m,2H),6.92(s,1H),6.78-6.70(m,1H),3.77( s,3H),3.30–3.22(m,1H),2.89–2.82(m,1H),2.00–1.91(m,2H),1.90–1.76(m,2H),1.60–1.46(m,3H), 1.44–1.31(m,5H).

Compound 28 (6.69 mg) was obtained, the second eluting isomer, as a white solid in 9% yield. (ESI): m/z 404.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.02(s, 1H), 8.87(s, 1H), 8.13–8.06(m, 1H) ),7.98(d,J＝11.0Hz,1H),7.67(t,J＝8.5Hz,1H),7.58(s,1H),7.39(d,J＝8.5Hz,1H),7.08(s,1H ),6.74(t,J＝10.0Hz,1H),3.76(s,3H),3.46-3.36(m,2H),2.15-2.07(m,1H),2.06-1.98(m,1H),1.92- 1.83 (m, 2H), 1.82–1.69 (m, 2H), 1.67–1.59 (m, 1H), 1.58–1.51 (m, 1H), 1.33 (d, J=6.5Hz, 3H), 1.22–1.14 ( m,1H).

Example 7: Compound 35 and Compound 36

Compound 35 (trans, racemic) and compound 36 (cis, racemic) proceeded from intermediate F (50 mg) and 4-bromo-1,2-phenylenediamine via general route one and general route three prepared.

Compound 35 (12.85 mg) was obtained, the first eluting isomer, as a white solid in 17% yield. (ESI): m/z 452.3 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.39 (d, J=22.9Hz, 1H), 8.80 (s, 1H), 8.10– 8.04(m, 1H), 7.98(d, J=11.0Hz, 1H), 7.74(s, 1H), 7.65(t, J=8.5Hz, 1H), 7.46–7.38(m, 2H), 7.26(t , J=9.0Hz, 1H), 3.31–3.24 (m, 1H), 2.95–2.87 (m, 1H), 1.99–1.90 (m, 2H), 1.89–1.79 (m, 2H), 1.61–1.46 (m ,3H),1.46–1.30(m,5H).

Compound 36 (15.29 mg) was obtained, the second eluting isomer, as a white solid in 20% yield. (ESI): m/z 452.3(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.42(s, 1H), 8.86(s, 1H), 8.12–8.07(m, 1H ),7.98(d,J＝11.0Hz,1H),7.73(s,1H),7.69-7.63(m,1H),7.60-7.56(m,1H),7.49(d,J＝9.0Hz,1H) ,7.28–7.22(m,1H),3.45–3.38(m,2H),2.15–2.08(m,1H),2.07–1.99(m,1H),1.92–1.83(m,2H),1.82–1.70( m, 2H), 1.68–1.59 (m, 1H), 1.58–1.52 (m, 1H), 1.34 (d, J=6.0Hz, 3H), 1.18–1.11 (m, 1H).

Example 8: Compound 37 and Compound 38

Compound 37 (trans, racemic) and compound 38 (cis, racemic) proceeded from intermediate F (40 mg) and 6-chloro-2,3-diaminopyridine via general route one and general route three prepared.

Compound 37 (4.03 mg) was obtained, the first eluting isomer, as a white solid in 7% yield. MS(ESI): m/z 409.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.79(s, 1H), 8.80(s, 1H), 8.11–8.04(m, 1H), 8.02–7.87 (m, 2H), 7.66 (t, J=8.0Hz, 1H), 7.44 (s, 1H), 7.24 (d, J=8.0Hz, 1H), 3.30–3.23 (m, 1H) ), 2.98–2.89 (m, 1H), 2.00–1.91 (m, 2H), 1.90–1.80 (m, 2H), 1.61–1.47 (m, 3H), 1.45–1.35 (m, 5H).

Compound 38 (7.12 mg) was obtained, the second eluting isomer, as a white solid in 12% yield. MS (ESI): m/z 409.4 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.81 (s, 1H), 8.87 (d, J=4.5Hz, 1H), 8.10 (dd,J=9.5,6.0Hz,1H),8.03–7.90(m,2H),7.67(td,J=9.0,3.0Hz,1H),7.59(d,J=4.5Hz,1H),7.23( d, J=8.5Hz, 1H), 3.47–3.39 (m, 2H), 2.18–2.12 (m, 1H), 2.07–2.00 (m, 1H), 1.92–1.83 (m, 2H), 1.82–1.71 ( m, 2H), 1.69–1.61 (m, 1H), 1.59–1.53 (m, 1H), 1.35 (d, J=7.0Hz, 3H), 1.17–1.11 (m, 1H).

Example 9: Compound 39 and Compound 40

Compound 39 (trans, racemic) and compound 40 (cis, racemic) proceeded from intermediate F (40 mg) and 5-chloro-2,3-diaminopyridine via general route one and general route three prepared.

Compound 39 (5.08 mg) was obtained, the first eluting isomer, as a white solid in 9% yield. MS(ESI): m/z 409.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.80(s, 1H), 8.81(s, 1H), 8.12–8.05(m, 1H), 8.04–7.88 (m, 2H), 7.65 (t, J=8.0Hz, 1H), 7.43 (s, 1H), 7.26 (d, J=8.0Hz, 1H), 3.31–3.23 (m, 1H) ), 2.99–2.88 (m, 1H), 2.02–1.90 (m, 2H), 1.93–1.81 (m, 2H), 1.60–1.46 (m, 3H), 1.46–1.34 (m, 5H).

Compound 40 (8.37 mg) was obtained, the second eluting isomer, as a white solid in 15% yield. MS(ESI): m/z 409.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.81(s, 1H), 8.81(s, 1H), 8.12–8.04(m, 1H), 8.03–7.87 (m, 2H), 7.64 (t, J=8.0Hz, 1H), 7.45 (s, 1H), 7.26 (d, J=8.0Hz, 1H), 3.46–3.37 (m, 2H) ), 2.15–2.07 (m, 1H), 2.05–1.98 (m, 1H), 1.93–1.85 (m, 2H), 1.84–1.68 (m, 2H), 1.67–1.60 (m, 1H), 1.57–1.50 (m, 1H), 1.36 (d, J=6.5Hz, 3H), 1.25–1.14 (m, 1H).

Example 10: Compound 41 and Compound 42

Compound 41 (trans, racemic) and compound 42 (cis, racemic) proceeded from intermediate F (40 mg) and 6-chloro-3,4-diaminopyridine via general route one and general route three prepared.

Compound 41 (2.98 mg) was obtained, the first eluting isomer, as a white solid in 5% yield. MS(ESI): m/z 409.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.89(s, 1H), 8.87(s, 1H), 8.63(s, 1H) ,8.14–8.08(m,1H),7.98(d,J=11.0Hz,1H),7.70–7.55(m,3H),3.48–3.40(m,1H),3.01–2.92(m,1H),2.05 –1.94(m,2H),1.96–1.85(m,2H),1.65–1.49(m,3H),1.48–1.38(m,5H).

Compound 42 (3.17 mg) was obtained, the second eluting isomer, as a white solid in 6% yield. MS(ESI): m/z 409.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.90(s, 1H), 8.87(s, 1H), 8.63(s, 1H) ,8.14–8.07(m,1H),7.99(d,J=11.0Hz,1H),7.71–7.56(m,3H),3.49–3.40(m,2H),2.18–2.11(m,1H),2.07 –2.01(m,1H),1.93-1.83(m,2H),1.82-1.72(m,2H),1.69-1.61(m,1H),1.60-1.53(m,1H),1.36(d,J= 6.0Hz, 3H), 1.17–1.10(m, 1H).

Example 11: Compound 29 and Compound 30

Compound 29 (trans, rac) and compound 30 (cis, rac) were prepared from intermediate F (40 mg) and 4-chloro-5-fluoro-1,2-phenylenediamine via general route one and general route three prepared.

Compound 29 (3.45 mg) was obtained, the first eluting isomer, as a white solid in 5% yield. MS (ESI): m/z 426.4 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.49 (s, 1H), 8.79 (d, J=4.5Hz, 1H), 8.10 –8.04(m,1H),7.97(d,J=11.0Hz,1H),7.68-7.57(m,2H),7.48(d,J=9.5Hz,1H),7.44-7.41(m,1H), 3.29-3.23(m, 1H), 2.94-2.88(m, 1H), 1.99-1.90(m, 2H), 1.88-1.79(m, 2H), 1.61-1.46(m, 3H), 1.44-1.34(m , 5H).

Compound 30 (4.15 mg) was obtained, the second eluting isomer, as a white solid in 6% yield. MS(ESI): m/z 426.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.54(s, 1H), 8.87(s, 1H), 8.13–8.06(m, 1H), 7.98(d, J＝11.0Hz, 1H), 7.78-7.63(s, 2H), 7.61-7.40(m, 2H), 3.45-3.39(m, 2H), 2.16-2.08(m, 1H) ,2.07–2.00(m,1H),1.92–1.83(m,2H),1.82–1.70(m,2H),1.68–1.60(m,1H),1.59–1.52(m,1H),1.34(d, J=6.5Hz, 3H), 1.18–1.11 (m, 1H).

Example 12: Compound 45 and Compound 46

Compound 45 (trans, racemic) and compound 46 (cis, racemic) were prepared from intermediate F (40 mg) and 2-amino-4-chlorophenol via general route one and general route three .

Compound 45 (6.36 mg) was obtained, the first eluting isomer, as a white solid in 12% yield. MS (ESI): m/z 409.4 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 8.87 (d, J=4.5Hz, 1H), 8.11 (dd, J=9.0, 6.0Hz, 1H), 8.00 (dd, J=11.0, 3.0Hz, 1H), 7.93 (d, J=2.0Hz, 1H), 7.74 (d, J=8.5Hz, 1H), 7.69 (td, J= 8.5, 3.0Hz, 1H), 7.65 (d, J=4.5Hz, 1H), 7.42 (dd, J=8.5, 2.0Hz, 1H), 3.65–3.56 (m, 1H), 3.48–3.44 (m, 1H) ), 2.17–2.11 (m, 1H), 2.05–1.98 (m, 1H), 1.92–1.71 (m, 5H), 1.67–1.61 (m, 1H), 1.39 (d, J=7.0Hz, 3H), 1.30–1.25(m,1H).

Compound 46 (9.27 mg) was obtained, the second eluting isomer, as a white solid in 17% yield. MS (ESI): m/z 409.4 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 8.86 (d, J=4.5Hz, 1H), 8.10 (dd, J=9.0, 6.0Hz, 1H), 8.01 (dd, J=11.0, 3.0Hz, 1H), 7.92 (d, J=2.0Hz, 1H), 7.72 (d, J=8.5Hz, 1H), 7.68 (td, J= 8.5, 3.0Hz, 1H), 7.63 (d, J=4.5Hz, 1H), 7.41 (dd, J=8.5, 2.0Hz, 1H), 3.64–3.50 (m, 2H), 2.17–2.11 (m, 1H) ), 2.05–1.98 (m, 1H), 1.92–1.71 (m, 5H), 1.67–1.61 (m, 1H), 1.39 (d, J=7.0Hz, 3H), 1.30–1.25 (m, 1H).

Example 13: Compound 43 and Compound 44

Compound 43 (trans, racemic) and compound 44 (cis, racemic) were prepared from intermediate F (40 mg) and 2-amino-5-chlorophenol via general route one and general route three .

Compound 43 (8.42 mg) was obtained, the first eluting isomer, as a white solid in 16% yield. MS (ESI): m/z 409.4 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 8.86 (d, J=4.5Hz, 1H), 8.10 (dd, J=9.0, 6.0Hz, 1H), 8.01 (dd, J=11.0, 3.0Hz, 1H), 7.92 (d, J=2.0Hz, 1H), 7.72 (d, J=8.5Hz, 1H), 7.68 (td, J= 8.5, 3.0Hz, 1H), 7.63 (d, J=4.5Hz, 1H), 7.41 (dd, J=8.5, 2.0Hz, 1H), 3.64–3.56 (m, 1H), 3.48–3.43 (m, 1H) ), 2.17–2.11 (m, 1H), 2.05–1.98 (m, 1H), 1.92–1.71 (m, 5H), 1.67–1.61 (m, 1H), 1.39 (d, J=7.0Hz, 3H), 1.30–1.25(m,1H).

Compound 44 (10.05 mg) was obtained, the second eluting isomer, as a white solid in 19% yield. MS (ESI): m/z 409.4 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 8.86 (d, J=4.5Hz, 1H), 8.10 (dd, J=9.0, 6.0Hz, 1H), 8.01 (dd, J=11.0, 3.0Hz, 1H), 7.92 (d, J=2.0Hz, 1H), 7.72 (d, J=8.5Hz, 1H), 7.68 (td, J= 8.5, 3.0Hz, 1H), 7.63 (d, J=4.5Hz, 1H), 7.41 (dd, J=8.5, 2.0Hz, 1H), 3.64–3.50 (m, 2H), 2.17–2.11 (m, 1H) ), 2.05–1.98 (m, 1H), 1.92–1.71 (m, 5H), 1.67–1.61 (m, 1H), 1.39 (d, J=7.0Hz, 3H), 1.30–1.25 (m, 1H).

Example 14: Compound 48

Compound 48 was prepared from intermediate H" (40 mg) and 4-methyl-1,2-phenylenediamine via general route four. Compound 48 (28.67 mg) was obtained as a white solid in 56% yield. ( ESI): m/z 386.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.32(s, 1H), 8.82(s, 1H), 8.11–8.04(m, 1H) ,8.00(d,J＝11.0Hz,1H),7.66(t,J＝8.5Hz,1H),7.39(s,1H),7.37–7.29(m,1H),7.29–7.19(m,1H), 6.91(d,J=8.0Hz,1H),3.42-3.37(m,1H),2.37(s,3H),2.28-2.20(m,1H),2.08-2.02(m,1H),1.95-1.86( m, 3H), 1.59–1.46 (m, 2H), 1.46–1.41 (m, 1H), 1.23 (d, J=13.5Hz, 1H), 1.15–1.03 (m, 2H).

Example 15: Compound 49

Compound 49 was prepared from intermediate I"' (40 mg) and 4-chloro-1,2-phenylenediamine via general route five. Compound 49 (15.23 mg) was obtained as a white solid in 28% yield. ( ESI): m/z 420.4(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.32(s, 1H), 8.82(s, 1H), 8.11–8.04(m, 1H) ,8.00(d,J＝11.0Hz,1H),7.66(t,J＝8.5Hz,1H),7.39(s,1H),7.37–7.29(m,1H),7.29–7.19(m,1H), 6.91(d,J=8.0Hz,1H),2.45(s,3H),3.42-3.37(m,1H),2.28-2.20(m,1H),2.08-2.02(m,1H),1.95-1.86( m, 3H), 1.59–1.46 (m, 2H), 1.46–1.41 (m, 1H), 1.15–1.03 (m, 2H).

Example 16: Compound 19 and Compound 20

Compound 19 and compound 20 were obtained from compound 18 in Example 2 through chiral column resolution. Among them, compound 19 corresponds to the former in chiral resolution, and compound 20 corresponds to the latter in chiral resolution.

Compound 19: MS (ESI): m/z 392.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.33 (s, 1H), 8.86 (s, 1H), 8.13–8.07 (m, 1H), 7.98(d, J=10.5Hz, 1H), 7.67(t, J=8.5Hz, 1H), 7.58(s, 1H), 7.44–7.39(m, 1H), 7.24(d, J=8.5Hz, 1H), 7.01–6.92 (m, 1H), 3.45–3.35 (m, 2H), 2.15–2.09 (m, 1H), 2.06–1.99 (m, 1H), 1.93–1.83 (m, 2H), 1.82–1.71 (m, 2H), 1.68–1.60 (m, 1H), 1.59–1.53 (m, 1H), 1.33 (d, J=6.0Hz, 3H), 1.21–1.14 (m, 1H) .

Compound 20: MS (ESI): m/z 392.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.34 (s, 1H), 8.87 (s, 1H), 8.13–8.07 (m, 1H), 7.98(d, J=10.5Hz, 1H), 7.67(t, J=8.5Hz, 1H), 7.58(s, 1H), 7.44–7.39(m, 1H), 7.24(d, J=8.5Hz, 1H), 7.01–6.92 (m, 1H), 3.45–3.35 (m, 2H), 2.15–2.09 (m, 1H), 2.06–1.99 (m, 1H), 1.93–1.83 (m, 2H), 1.82–1.71 (m, 2H), 1.68–1.60 (m, 1H), 1.59–1.53 (m, 1H), 1.34 (d, J=6.5Hz, 3H), 1.21–1.14 (m, 1H) .

Example 17: Compound 23 and Compound 24

Compound 23 and compound 24 were obtained from compound 22 in Example 3 through chiral column resolution. Among them, compound 23 corresponds to the former in chiral resolution, and compound 24 corresponds to the latter in chiral resolution.

Compound 23: MS (ESI): m/z 388.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.03 (s, 1H), 8.86 (s, 1H), 8.13–8.06 (m, 1H), 7.97(d, J=10.5Hz, 1H), 7.66(t, J=9.0Hz, 1H), 7.58(s, 1H), 7.41–7.18(m, 2H), 6.93(s, 1H), 3.46–3.34 (m, 2H), 2.38 (s, 3H), 2.16–2.09 (m, 1H), 2.06–1.99 (m, 1H), 1.93–1.83 (m, 2H), 1.82–1.70 ( m, 2H), 1.66–1.58 (m, 1H), 1.57–1.50 (m, 1H), 1.33 (d, J=6.5Hz, 3H), 1.21–1.14 (m, 1H).

Compound 24: MS (ESI): m/z 388.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.07 (s, 1H), 8.87 (s, 1H), 8.13–8.06 (m, 1H), 7.97(d, J=10.5Hz, 1H), 7.66(t, J=9.0Hz, 1H), 7.58(s, 1H), 7.41–7.18(m, 2H), 6.93(s, 1H), 3.46–3.34 (m, 2H), 2.38 (s, 3H), 2.16–2.09 (m, 1H), 2.06–1.99 (m, 1H), 1.93–1.83 (m, 2H), 1.82–1.70 ( m, 2H), 1.66–1.58 (m, 1H), 1.57–1.50 (m, 1H), 1.33 (d, J=6.5Hz, 3H), 1.21–1.14 (m, 1H).

Example 18: Compound 15 and Compound 16

Compound 15 and compound 16 were obtained from compound 14 in Example 4 through chiral column resolution. Among them, compound 15 corresponds to the former in chiral resolution, and compound 16 corresponds to the latter in chiral resolution.

Compound 15: MS (ESI): m/z 408.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.41 (s, 1H), 8.87 (s, 1H), 8.12–8.06 (m, 1H), 7.98 (d, J=10.5Hz, 1H), 7.67 (t, J=8.5Hz, 1H), 7.61–7.56 (m, 2H), 7.46 (d, J=8.5Hz, 1H) ,7.17–7.11(m,1H),3.45–3.37(m,2H),2.16–2.09(m,1H),2.07–2.00(m,1H),1.93–1.83(m,2H),1.82–1.71( m, 2H), 1.68–1.60 (m, 1H), 1.59–1.52 (m, 1H), 1.35 (d, J=6.5Hz, 3H), 1.19–1.12 (m, 1H).

Compound 16: MS (ESI): m/z 408.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.46 (s, 1H), 8.87 (s, 1H), 8.12–8.06 (m, 1H), 7.98 (d, J=10.5Hz, 1H), 7.67 (t, J=8.5Hz, 1H), 7.62–7.40 (m, 3H), 7.14 (d, J=8.5Hz, 1H) ,3.45–3.37(m,2H),2.16–2.09(m,1H),2.07–2.00(m,1H),1.93–1.83(m,2H),1.82–1.71(m,2H),1.68–1.60( m, 1H), 1.59–1.52 (m, 1H), 1.35 (d, J=6.5Hz, 3H), 1.19–1.12 (m, 1H).

Example 19: Compound 31 and Compound 32

Compound 31 and compound 32 were obtained from compound 30 in Example 11 through chiral column resolution. Among them, compound 31 corresponds to the former in chiral resolution, and compound 32 corresponds to the latter in chiral resolution.

Compound 31: MS (ESI): m/z 426.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.55 (s, 1H), 8.87 (s, 1H), 8.13–8.06 (m, 1H), 7.98 (d, J=11.0Hz, 1H), 7.78–7.63 (s, 2H), 7.61–7.40 (m, 2H), 3.45–3.39 (m, 2H), 2.16–2.08 (m ,1H),2.07–2.00(m,1H),1.92–1.83(m,2H),1.82–1.70(m,2H),1.68–1.60(m,1H),1.59–1.52(m,1H),1.34 (d, J=6.5Hz, 3H), 1.18–1.11 (m, 1H).

Compound 32: MS (ESI): m/z 426.4 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.54 (s, 1H), 8.86 (s, 1H), 8.13–8.06 (m, 1H), 7.98 (d, J=11.0Hz, 1H), 7.78–7.63 (s, 2H), 7.61–7.40 (m, 2H), 3.45–3.39 (m, 2H), 2.16–2.08 (m ,1H),2.07–2.00(m,1H),1.92–1.83(m,2H),1.82–1.70(m,2H),1.68–1.60(m,1H),1.59–1.52(m,1H),1.34 (d, J=6.5Hz, 3H), 1.18–1.11 (m, 1H).

Example 20: Compound 51 and Compound 52

Compound 51 (trans, rac) and compound 52 (cis, rac) were prepared from intermediate F (40 mg) and 4,5-difluoro-1,2-phenylenediamine via general routes one and Prepared by general route three.

Compound 51 (3.45 mg) was obtained, the first eluting isomer, as a white solid in 5% yield. MS (ESI): m/z 410.2 (M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.49 (s, 1H), 8.79 (d, J=4.5Hz, 1H), 8.10 –8.04(m,1H),7.97(d,J=11.0Hz,1H),7.68-7.57(m,2H),7.48(d,J=9.5Hz,1H),7.44-7.41(m,1H), 3.29-3.23(m, 1H), 2.94-2.88(m, 1H), 1.99-1.90(m, 2H), 1.88-1.79(m, 2H), 1.61-1.46(m, 3H), 1.44-1.34(m , 5H).

Compound 52 (4.15 mg) was obtained, the second eluting isomer, as a white solid in 6% yield. MS(ESI): m/z 410.2(M+H) ⁺ . ¹ H NMR (500MHz, d ₆ -DMSO) δ 12.54(s, 1H), 8.87(s, 1H), 8.13–8.06(m, 1H), 7.98(d, J＝11.0Hz, 1H), 7.78-7.63(s, 2H), 7.61-7.40(m, 2H), 3.45-3.39(m, 2H), 2.16-2.08(m, 1H) ,2.07–2.00(m,1H),1.92–1.83(m,2H),1.82–1.70(m,2H),1.68–1.60(m,1H),1.59–1.52(m,1H),1.34(d, J=6.5Hz, 3H), 1.18–1.11 (m, 1H).

Example 21: Compound 53 and Compound 54

Compound 53 and compound 54 were obtained from compound 52 in Example 21 through chiral column resolution. Among them, compound 53 corresponds to the former in chiral resolution, and compound 54 corresponds to the latter in chiral resolution.

Compound 53: MS (ESI): m/z 410.2 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.54 (s, 1H), 8.87 (s, 1H), 8.13–8.06 (m, 1H), 7.98 (d, J=11.0Hz, 1H), 7.78–7.63 (s, 2H), 7.61–7.40 (m, 2H), 3.45–3.39 (m, 2H), 2.16–2.08 (m ,1H),2.07–2.00(m,1H),1.92–1.83(m,2H),1.82–1.70(m,2H),1.68–1.60(m,1H),1.59–1.52(m,1H),1.34 (d, J=6.5Hz, 3H), 1.18–1.11 (m, 1H).

Compound 54: MS (ESI): m/z 410.2 (M+H) ⁺ . ¹ H NMR (500 MHz, d ₆ -DMSO) δ 12.54 (s, 1H), 8.87 (s, 1H), 8.13–8.06 (m, 1H), 7.98 (d, J=11.0Hz, 1H), 7.78–7.63 (s, 2H), 7.61–7.40 (m, 2H), 3.45–3.39 (m, 2H), 2.16–2.08 (m ,1H),2.07–2.00(m,1H),1.92–1.83(m,2H),1.82–1.70(m,2H),1.68–1.60(m,1H),1.59–1.52(m,1H),1.34 (d, J=6.5Hz, 3H), 1.18–1.11 (m, 1H).

Claims (2)

1. A process for the preparation of a compound of formula a, characterized by the steps of:

the first step is as follows: dissolving triethyl phosphonoacetate in ultra-dry tetrahydrofuran, and adding sodium tert-butoxide at 0 ℃ in an ice bath; adding the tetrahydrofuran solution of the intermediate E' into the reaction solution; after 2 hours of reaction, quench with water; extracting the aqueous solution with 20mL ethyl acetate for three times, combining organic phases, washing with saturated salt water, drying with anhydrous sodium sulfate, filtering, concentrating, and separating the residue with a rapid column chromatography to obtain an intermediate F ";

the second step is that: NaH was added to 15mL of dimethyl sulfoxide, trimethyl sulfoxide iodide was added to the suspension, the mixture was stirred at room temperature for 1.5 hours, and then a dimethyl sulfoxide solution of intermediate F' was added to the reaction solution. The reaction is stirred overnight at room temperature, then quenched with water, extracted with ethyl acetate and separated by a fast column chromatography to obtain an intermediate G ";

the third step: dissolving the intermediate G 'in 10mL of ethanol, adding 4mL of 2mol/L sodium hydroxide solution, heating the reaction solution to 50 ℃, reacting for 2 hours, cooling the reaction solution to room temperature, neutralizing the reaction solution with 4mol/L hydrochloric acid solution until the pH value is 1, extracting the water phase with ethyl acetate, combining the organic phases, drying with anhydrous sodium sulfate, filtering, concentrating, and separating the residue by preparative thin-layer chromatography to obtain an intermediate H';

the fourth step: dissolving the intermediate H' in N, N-dimethylformamide, adding HATU and diisopropylethylamine, adding the substituted 1, 2-diamine to the reaction solution, stirring the reaction mixture at 30 ℃ overnight, adding water and ethyl acetate to the reaction solution, washing the organic phase with saturated saline, drying over anhydrous sodium sulfate, filtering, and concentrating. The crude intermediate obtained was dissolved in acetic acid, the mixture was stirred at 100 ℃ for reaction for 19 hours, then the reaction solution was concentrated, and the residue was purified by reverse phase high performance liquid chromatography to give the final compound.

2. The method of claim 1 wherein the substituted o-amino aniline is selected from the group consisting of 4-methyl-1, 2-phenylenediamine.