CN105294669B

CN105294669B - A kind of factor X inhibitor and its preparation method and application

Info

Publication number: CN105294669B
Application number: CN201510697138.6A
Authority: CN
Inventors: 吴俊军; 李锐; 姜祥胜; 宋嘉栖; 杨双兵; 张璟煜; 张二利
Original assignee: Shandong Kaisen Pharma Co Ltd
Current assignee: Shandong Kaisen Pharma Co Ltd
Priority date: 2014-10-24
Filing date: 2015-10-23
Publication date: 2019-01-22
Anticipated expiration: 2035-10-23
Also published as: CN105294669A

Abstract

The present invention relates to a kind of factor X inhibitor and its preparation method and application, the factor X inhibitor has the structure of general formula I, inhibitor of the present invention is using alpha- amino acid as template, form a series of compound of structure novels for branch by amide, carbamate or urea respectively, this kind of compound can prevent the formation of thrombus effectively in conjunction with factor X.

Description

A kind of factor X inhibitor and its preparation method and application

Technical field

The present invention relates to field of medicine invention, and in particular to a kind of factor X (FXa) inhibitor and preparation method thereof and Using.

Background technique

Thrombus is the blood clotting or deposit that blood constituent is formed in blood vessel or cardiac intima surface, it can occur From anywhere in blood vessel, blood flow is caused to stop.From the beginning foot is arrived in the performance of thrombotic disease has, cerebrovascular thrombus Patient can be made side limbs disturbance, i.e. hemiplegia, aphasia, vision and sensory disturbance, stupor occur, can disable, is lethal.Heart The thrombosis of blood vessel, i.e. shape Arterial thrombosis, can lead to serious angina pectoris or even myocardial infarction.According to world health group (WHO) statistics is knitted, there are about 1,2,000,000 people to die of cerebral thrombosis, cerebral infarction, myocardial infarction, coronary heart disease, artery sclerosis etc. every year in the whole world Disease, close to the 1/4 of the total death toll in the world.

Antithrombotic reagent mainly includes anticoagulant, platelet aggregation inhibitor and thrombolytic, wherein anticoagulation medicine and Medicament for resisting platelet aggregation clinically has wide application.In anticoagulation medicine, warfarin (walfrin), heparin (heprin) it has dominated over half a century with low molecular weight heparin and has occupied a tiny space in antithrombotic reagent.Antiplatelet is poly- The drug of collection has aspirin (aspirin) and clopidogrel (clopidogrel), these most common antithrombotic reagents are maximum Side effect be to cause bleeding, wherein aspirin and clopidogrel such as dosage deficiency also result in recurrent thrombus and are formed.Liver The thrombopenia of element induction and warfarin bring cutaneous necrosis etc. are more that warfarin treatment window narrows work Slowly, more frequently blood monitoring is needed to face it in the presence of interaction between a variety of foods or drug to adjust dosage and it It is restricted in bed application.

New drug target is based primarily upon searching and participates in blood clotting cascade reaction (coagulation cascade) various factors Inhibitor on, factor X is located at the intersection of inherent approach and external approach in coagulation cascade reaction, inhibits factor X On the one hand it can effectively inhibit a large amount of formation of fresh blood clotting enzymes factor and coagulation cascade is blocked to react, on the other hand because it is not interfered Already existing blood clotting enzymes factor in blood and reduce bleeding risk.

In recent years, exploitation focus gradually focuses on factor X, and the factor X inhibitor developed has:

Benzamidine derivative DX-9065a (Nagahara T, the Yokoyama Y, Inamura of Daiichi pharmaceutical companies K,et al.Dibasic(amidinoaryl)propanoic acid derivatives as novel blood coagulation Factor Xa inhibitors.J Med Chem 1994；37:1200-7)；

Otamixaban (Guertin KR, Gardner CI, the Klein SI, et of Sanofi-Aventis pharmaceutical companies al.Optimization of the beta-aminoester class of Factor Xa inhibitors.Part 2: identifi cation of FXV673as a potent and selective inhibitor with excellent in vivo anticoagulant activity.Bioorg Med Chem Lett 2002；12:1671-4)；

Razaxaban (Quan MI, Lam PY, Han Q, the et al.Discovery of1- of DuPont pharmaceutical companies (3′-aminobenzisoxazol-′-yl)-3-trifluoromethyl-N-[2-fluoro-4-[(2′- dimethylaminomethyl)imidazol-1-yl]p henyl]-1H-pyrazole-5-carboxamide hydrochloride(razaxaban),a highly potent,selective,and orally biovailable Factor Xa inhibitor.J Med Chem 2005；48:1729-44)；

In recent years the antithrombotic reagent for going through to list has:

Razaxaban (rivaroxaban, BAY 59-7939) (Roherig S, Straub A, Pohlmann J, et al.Discovery of the novel antithrombotic agent 5-chloro-N-({(5S)-2-oxo-3-[4- (3-oxomorpholin-4-yl)phenyl]-1,3-oxazolidin-5-yl}methyl)thiophene-2- carboxamide(BAY 59-7939):an oral,direct factor Xa inhibitor.J Med Chem 2005； 48:5900-8)；

Eliquis (Apixaban, BMS-562247-1) (Pinto DJ, Orwat MJ, Koch S, et al.Discovery of1-(4-methoxyphenyl)-7-oxo-6-(4-(2-oxopiperidin-1-yl)phenyl)-4, 5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide(Apixaban,BMS- 562247),a highly potent,selective,effi cacious,and orally bioavailable inhibitor of blood coagulation factor Xa.J Med Chem 2007；50:5339-56) etc..

A new class of factor X inhibitor provided by the invention has structure novel, inhibitory activity high, selective good Advantage.

Summary of the invention

The object of the present invention is to provide a kind of new factor X inhibitor.

The compound or pharmaceutically acceptable salt with logical structure shown in formula I provided by the invention:

Wherein,

R₁For aryl, heterocyclic group, alkyl, aryl ether, heterocyclic ether, alkyl ether, arylamine group, heterocycle amido or alkylamine Base；

R₂For aryl, heterocyclic group or alkyl；

R₃For morpholine ketone phenyl, aryl, heterocyclic group or alkyl；

X is nitrogen.

The heterocyclic group includes the groups such as pyridine, pyrimidine, thiazole, thiophene, oxazole, imidazoles, pyrazoles, pyrroles or indoles.

Preferably, the compound is the compound having following structure:

The present invention also provides the preparation methods of above compound as shown in Scheme 1, method includes the following steps: By the a-amino acid of N-protected and corresponding R₃-NH₂Reaction obtains corresponding amide, removes protective agent in acid condition, then with Carboxylic acid or the reactions such as acyl chlorides condensation or isocyanates, chloro-formate obtain corresponding factor X inhibitor compound.

The synthesis and preparation of 1 general formula I type compound of Scheme

The present invention also provides the preparation containing above compound, the preparation is by compound and pharmaceutically acceptable carrier Composition.

The preparation is tablet, capsule, granule.

The pharmaceutically acceptable carrier is selected from filler, disintegrating agent, adhesive, lubricant, including but not limited to appoints What and whole solvent, decentralized medium, coating, absorption delaying agent etc., such medium and medicament exist for pharmaceutically active substances This field is well-known.

The present invention also provides the preparation method of above-mentioned preparation, the routine side that the method is known to the skilled person Method.

The present invention also provides the compounds or preparation to prepare the application in the drug for treating and preventing thrombosis, The thrombus is that stroke or systemic embolism (SE), prevention hip joint and knee prosthesis occur for non-valvular atrial fibrillation (AF) patient Postoperative patient deep vein thrombosis (DVT) and pulmonary embolism (PE).

Compound provided by the invention has the advantage that

1, inhibitor of the present invention is respectively branch by amide, carbamate or urea using alpha- amino acid as template Chain forms a series of compound of structure novels, and this kind of compound can prevent the shape of thrombus effectively in conjunction with factor X At.

2, compound provided by the invention can selectively inhibit the tenth inhibition, and factor X is located in coagulation cascade reaction The intersection (Fig. 1) of inherent approach and external approach inhibits FXa, on the one hand can effectively inhibit a large amount of of fresh blood clotting enzymes factor It is formed and coagulation cascade is blocked to react, on the other hand reduce bleeding due to it does not interfere already existing blood clotting enzymes factor in blood Risk, therefore factor X inhibitor a kind of in this way can become a kind of and be used for anticoagulant drug.

Detailed description of the invention

Fig. 1: clotting mechanism schematic diagram.

Specific embodiment

The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..

Embodiment 1:(S)-N- (5-guanidino-1-oxo-1- (4- (3-oxomorpholino) phenylamino) Pentan-2-yl) the preparation of -6-methoxynico-tinamide (FXa-01)

1, intermediate (S)-tert-butyl-5- (3-nitroguanidino) -1-oxo-1- (4- (3- is prepared oxomorpholino)-phenylamino)pentan-2-ylcarbamate

Starting material 4- (4-aminophenyl) morpholin-3-one (250mg, 1.30mmol) is dissolved in N, N- bis- In methylformamide (1ml) and tetrahydrofuran (15ml), N, N- diisopropyl ethyl amine (0.7ml, 3.9mmol) are added under ice bath (S) -2- (tert-butoxycarbonyl) -5- (3-nitroguanidino) pentanoic acid (424mg, 1.33mmol), 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) are eventually adding (593mg, 1.56mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 2 hours later.LCMS detects fully reacting, A small amount of water quenching is added to go out, is extracted with ethyl acetate three times, combined organic phase saturated common salt water washing, anhydrous sodium sulfate is done It is dry, concentration.Gained residue obtains yellow solid, 350mg is pure through column chromatographic purifying (methylene chloride: methanol=80:1~20:1) Degree: 95%, yield: 45%.LC-MS(ESI):m/z 494(M+H)⁺。

2, intermediate (S) -2-amino-5- (3-nitroguanidino)-N- (4- (3-oxomorpholino) is prepared phenyl)-pentanamide

Step 1 products therefrom (350mg, 0.71mmol) is dissolved in methylene chloride (2ml), trifluoroacetic acid is added dropwise under ice bath (1ml) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 4 hours later.LCMS detects fully reacting.Reaction solution is set In being concentrated to dryness in revolving, then with vacuum drying, brown solid is obtained, 280mg, purity: 92%, yield: 100%.Gained is remaining Object is directly used in react in next step.LC-MS(ESI):m/z 394(M+H)⁺。

3, intermediate (S) -6-methoxy-N- (5- (3-nitroguanidino) -1-oxo-1- (4- (3- is prepared oxomorpholino)-phenylamino)pentan-2-yl)nicotinamide

Step 2 products therefrom (280mg, 0.71mmol) is dissolved in n,N-Dimethylformamide (1ml) and tetrahydrofuran In (15ml), N, N- diisopropyl ethyl amine (0.37ml, 2.13mmol) and 6-methoxynicotinic are added under ice bath Acid (130mg, 0.85mmol) is eventually adding 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid Ester (HATU) (323mg, 0.85mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 2 hours later.LCMS detection Fully reacting is added a small amount of water quenching and goes out, and is extracted with ethyl acetate three times, and combined organic phase saturated common salt water washing is anhydrous Sodium sulphate is dry, concentration.Gained residue obtains brown solid through column chromatographic purifying (methylene chloride: methanol=80:1~20:1), 130mg, purity: 92%, yield: 35%.LC-MS(ESI):m/z 529(M+H)⁺。

4, (S)-N- (5-guanidino-1-oxo-1- (4- (3-oxomorpholino) phenylamino) is prepared pentan-2-yl)-6-methoxynico-tinamide(FXa-1)

Step 3 products therefrom (110mg, 0.21mmol) is dissolved in the hydrogenation bottle equipped with dehydrated alcohol, palladium carbon is added (30mg) three times with hydrogen displacement is forced into 50psi, waves hydrogenation at room temperature 6 hours.LCMS detects fully reacting, Palladium carbon is filtered off, gained filtrate decompression is concentrated to dryness, and gained residue is crystallized through methanol and petroleum ether (1:15), and it is solid to obtain off-white color Body, 80mg, purity: 99%, yield: 80%.LC-MS(ESI):m/z 484(M+H)⁺；¹H NMR(400MHz,MeOD)δppm 1.6–1.9(m,2H),1.9–2.2(m,2H),3.1–3.4(m,3H),3.7–3.8(t,2H),3.9–4.0(s,3H),4.0–4.1 (t, 2H), 4.2-4.3 (s, 2H), 4.7-4.8 (m, 1H), 6.8-6.9 (d, 1H, J=8.8Hz), 7.3-7.4 (d, 2H, J= 8.8Hz), 7.7-7.8 (d, 2H, J=8.8Hz), 8.2-8.3 (m, 1H)；8.5-8.6(s；1H)；8.7-8.8 (d, 1H, J= 2.4Hz)。

Embodiment 2:(2S, 4R) -1- (2-chlorothiophene-5-carbonyl) -4-methoxy-N- (quinolin-2-yl) preparation of pyrrolidine-2-carboxamide (FXa-03)

1, intermediate (2S, 4R)-tert-butyl 4-methoxy-2- (quinolin-2-ylcarbamoyl) is prepared pyrrolidine-1-carboxylate

Quinolin-2-amine (144mg, 1.0mmol) is dissolved in N,N-dimethylformamide (0.5ml) and tetrahydro furan It mutters in (5ml), N, N- diisopropyl ethyl amine (0.53ml, 3.0mmol) and (2S, 4R) -1- (tert- is added under ice bath Butoxycarbonyl) -4-methoxypyrrolidine-2-carboxylic acid (245mg, 1.0mmol), finally plus Enter 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) (456mg, 1.2mmol), ice bath Under be stirred to react 15 minutes, go to later room temperature reaction 2 hours.LCMS detects fully reacting, and a small amount of water quenching is added and goes out, uses acetic acid Ethyl ester extracts three times, combined organic phase saturated common salt water washing, and anhydrous sodium sulfate is dry, concentration.Gained residue is through column Chromatographic purifying (petroleum ether: ethyl acetate=8:1~3:1), obtains off-white powder, 370mg, yield: 100%.LC-MS(ESI): m/z 372(M+H)⁺。

2, intermediate (2S, 4R)-tert-butyl 4-methoxy-2- (quinolin-2-ylcarbamoyl) is prepared pyrrolidine-1-carboxylate

Step 1 products therefrom is dissolved in methylene chloride (2ml), tetrahydrofuran (1ml) is added dropwise under ice bath, is stirred under ice bath Reaction 15 minutes goes to room temperature reaction 3 hours later.LCMS detects fully reacting.It places reaction liquid into revolving and is concentrated to dryness, Again with vacuum drying, yellow oily liquid is obtained, 70mg, purity: 92%, yield: 96%.Gained residue is directly used in next step Reaction.LC-MS(ESI):m/z272(M+H)⁺。

3, (2S, 4R) -1- (2-chlorothiophene-5-carbonyl) -4-methoxy-N- (quinolin- is prepared 2-yl)-pyrrolidine-2-carboxamide(FXa-2)

Step 2 products therefrom (70mg, 0.26mmol) is dissolved in N,N-dimethylformamide (0.5ml) and tetrahydrofuran In (3ml), it is added N under ice bath, N- diisopropyl ethyl amine (100mg, 0.77mmol) and 5- chlorothiophene -2- carboxylic acid (46mg, 0.28mmol), 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) are eventually adding (118mg, 0.31mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 2 hours later.LCMS detects fully reacting, A small amount of water quenching is added to go out, is extracted with ethyl acetate three times, combined organic phase saturated common salt water washing, anhydrous sodium sulfate is done It is dry, concentration.Gained residue obtains yellow solid, 18mg is pure through column chromatographic purifying (petroleum ether: ethyl acetate=8:1~3:1) Degree: 95%, yield: 17%.LC-MS(ESI):m/z 416(M+H)⁺；¹H NMR(400MHz,MeOD)δppm 2.12–2.22 (m,1H),2.52–2.62(m,1H),3.31–3.36(s,3H),3.9-4.2(m,2H),4.2-4.3(m,1H),4.4-4.6(m, 2H), 7.07-7.08 (d, 1H, J=3.6Hz), 7.48-7.50 (m, 1H), 7.54-7.55 (d, 1H, J=3.6Hz), 7.68 (t, 1H).7.83-7.86 (d, 2H, J=8.4Hz), 8.26 (S, 2H).

Embodiment 3:(S)-N- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -5-chlorothiophene-2-carboxamide (FXa-04) preparation

1, intermediate (S)-tert-butyl-3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenyl-amino)propan-2-ylcarbamate

4- (4-aminophenyl) morpholin-3-one (600mg, 3.12mmol) is dissolved in N, N- dimethyl formyl In amine (3ml) and tetrahydrofuran (30ml), N, N- diisopropyl ethyl amine (0.81ml, 4.68mmol) and Boc- are added under ice bath L-Histidine (798mg, 3.12mmol) is eventually adding 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea six Fluorophosphoric acid ester (HATU) (1.425g, 3.75mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 2 hours later. LCMS detects fully reacting, and a small amount of water quenching is added and goes out, is extracted with ethyl acetate three times, combined organic phase is washed with saturated common salt It washs, anhydrous sodium sulfate is dry, concentration.Gained residue obtains yellow through column chromatographic purifying (methylene chloride: methanol=50:1~10:1) Color solid, 480mg, purity: 98%, yield: 36%.LC-MS(ESI):m/z 430(M+H)⁺。

2, intermediate (S) -2-amino-3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino) is prepared phenyl)-propanamide

Step 1 products therefrom (160mg, 0.37mmol) is dissolved in methylene chloride (2ml), trifluoroacetic acid is added dropwise under ice bath (1ml) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 4 hours later.LCMS detects fully reacting.Reaction solution is set In being concentrated to dryness in revolving, then with vacuum drying, yellow solid is obtained, 123mg, purity: 98%, yield: 99%.Gained residue It is directly used in and reacts in next step.LC-MS(ESI):m/z 330(M+H)⁺。

3, (S)-N- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) phenyl- is prepared amino)propan-2-yl)-5-chlorothiophene-2-carboxamide

Step 2 products therefrom (123mg, 0.37mmol) is dissolved in N,N-dimethylformamide (0.5ml) and tetrahydrofuran In (10ml), it is added N under ice bath, N- diisopropyl ethyl amine (0.16ml, 0.94mmol) and 5- chlorothiophene -2- carboxylic acid (67mg, 0.41mmol), 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) are eventually adding (171mg, 0.45mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later.LCMS detects fully reacting, A small amount of water quenching is added to go out, is extracted with ethyl acetate three times, combined organic phase saturated common salt water washing, anhydrous sodium sulfate is done It is dry, concentration.Gained residue obtains off-white powder through column chromatographic purifying (methylene chloride: methanol=80:1~15:1), 86mg, Purity: 95%, yield: 49%.LC-MS(ESI):m/z 474(M+H)⁺；¹H NMR(400MHz,MeOD)δppm 3.1–3.2 (m,1H),3.2–3.3(m,1H),3.7-3.8(m,2H),4.0–4.1(m,2H),4.28(s,2H),6.92(s,1H),7.04– 7.05 (d, 1H, J=4Hz), 7.31-7.33 (d, 2H, J=8Hz), 7.62-7.65 (m, 4H).

Embodiment 4:(S)-N- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -4-chlorobenzamide (FXa-05) preparation

By (S) -2-amino-3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino) phenyl) Propanamide (100mg, 0.304mmol) is dissolved in n,N-Dimethylformamide (1ml) and tetrahydrofuran (5ml), under ice bath N, N- diisopropyl ethyl amine (0.16ml, 0.94mmol) and parachlorobenzoic-acid (67mg, 0.41mmol) is added, is eventually adding 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) (171mg, 0.45mmol), under ice bath It is stirred to react 15 minutes, goes to room temperature reaction 3 hours later.LCMS detects fully reacting, and a small amount of water quenching is added and goes out, with acetic acid second Ester extracts three times, combined organic phase saturated common salt water washing, and anhydrous sodium sulfate is dry, concentration.Gained residue is through column layer Analysis purifying (methylene chloride: methanol=80:1~15:1), obtains off-white powder, 82mg, purity: 95%, yield: 58%.LC-MS (ESI):m/z 468(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 3.0–3.1(m,1H),3.1–3.2(m,1H),3.8- 3.9 (m, 2H), 4.1-4.2 (m, 2H), 4.18 (s, 2H), 6.97 (S, 1H), 7.30-7.32 (d, 2H, J=8Hz), 7.55- 7.57 (d, 2H, J=8Hz), 7.60-7.63 (d, 2H, J=12Hz), 7.82 (S, 1H), 7.89-7.91 (d, 2H, J= 8.8Hz), 8.85-8.87 (d, 1H, J=8Hz), 10.21 (S, 1H).

Embodiment 5:(S)-N- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -4-chlorobenzamide (FXa-06) preparation

1, intermediate (S)-tert-butyl-3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenylamino)propan-2-ylcarbamate

4- (4-aminophenyl) morpholin-3-one (400mg, 2.08mmol) is dissolved in N, N- dimethyl formyl In amine (2ml) and tetrahydrofuran (20ml), N, N- diisopropyl ethyl amine (0.54ml, 3.2mmol) and Boc- are added under ice bath L-Tryptophan-OH (586mg, 2.08mmol) is eventually adding 2- (7- azo benzotriazole)-N, N, N', N'- tetramethyl Urea hexafluorophosphoric acid ester (HATU) (950mg, 2.5mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 2 hours later. LCMS detects fully reacting, and a small amount of water quenching is added and goes out, is extracted with ethyl acetate three times, combined organic phase is washed with saturated common salt It washs, anhydrous sodium sulfate is dry, concentration.Gained residue obtains class through column chromatographic purifying (methylene chloride: methanol=50:1~10:1) White solid, 375mg, purity: 98%, yield: 40%.LC-MS(ESI):m/z456(M+H)⁺。

2, intermediate (S) -2-amino-3- (4-hydroxyphenyl)-N- (4- (3-oxomorpholino) is prepared phenyl)propanamide

Step 1 products therefrom (360mg, 0.79mmol) is dissolved in methylene chloride (2ml), trifluoroacetic acid is added dropwise under ice bath (1ml) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 4 hours later.LCMS detects fully reacting.Reaction solution is set In being concentrated to dryness in revolving, then with vacuum drying, brown solid is obtained, 280mg, purity: 98%, yield: 99%.Gained residue It is directly used in and reacts in next step.LC-MS(ESI):m/z 356(M+H)⁺。

3, (S) -5-chloro-N- (3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenylamino)propan-2-yl)thiophene-2-carboxamide

Step 2 products therefrom (260mg, 0.73mmol) is dissolved in n,N-Dimethylformamide (1ml) and tetrahydrofuran In (10ml), it is added N under ice bath, N- diisopropyl ethyl amine (284mg, 2.20mmol) and 5- chlorothiophene -2- carboxylic acid (120mg, 0.73mmol), 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) are eventually adding (334mg, 0.88mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 2 hours later.LCMS detects fully reacting, A small amount of water quenching is added to go out, is extracted with ethyl acetate three times, combined organic phase saturated common salt water washing, anhydrous sodium sulfate is done It is dry, concentration.Gained residue obtains off-white powder through column chromatographic purifying (methylene chloride: methanol=50:1~10:1), 70mg, Purity: 96%, yield: 19%.LC-MS(ESI):m/z 500(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 2.8–2.9 (m,2H),3.0–3.1(m,1H),3.5–3.6(m,1H),3.6–3.7(m,2H),3.9–4.0(m,2H),4.18(S,2H), 4.6-4.7 (m, 1H), 6.63-6.65 (d, 4H, J=8.4Hz), 7.13-7.16 (d, 2H, J=8.4Hz), 7.18-7.20 (d, 1H, J=4Hz), 7.31-7.34 (d, 2H, J=8.8Hz), 7.60-7.63 (d, 2H, J=8.8HZ), 7.79-7.81 (d, 1H, J =4Hz), 8.86-8.89 (d, 1H, J=8.4Hz), 10.28 (s, 1H).

Embodiment 6:(R)-N- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -5-chlorothiophene-2-carboxamide (FXa-14) preparation

1, intermediate (R)-tert-butyl-3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3- is prepared oxomorpholino)-phenylamino)-propan-2-ylcarbamate

4- (4-aminophenyl) morpholin-3-one (2.0g, 10.41mmol) is dissolved in N, N- dimethyl formyl In amine (10ml) and tetrahydrofuran (20ml), N, N- diisopropyl ethyl amine (5.4g, 41.64mmol) and Boc- are added under ice bath D-Histidine-OH (2.66g, 10.41mmol) is eventually adding 2- (7- azo benzotriazole)-N, N, N', N'- tetramethyl Urea hexafluorophosphoric acid ester (HATU) (4.75g, 12.49mmol) is stirred to react 15 minutes under ice bath, and it is small to go to room temperature reaction 3 later When.LCMS detects fully reacting, and a small amount of water quenching is added and goes out, is extracted with ethyl acetate three times, combined organic phase saturated common salt Water washing, anhydrous sodium sulfate is dry, concentration.Gained residue through column chromatographic purifying (methylene chloride: methanol=50:1~15:1), Off-white powder, 1.21g, purity: 94%, yield: 27%.LC-MS(ESI):m/z 430(M+H)⁺。

2, intermediate (R) -2-amino-3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino) is prepared phenyl)propanamide

Step 1 products therefrom (1.20g, 2.79mmol) is dissolved in methylene chloride (10ml), trifluoro second is added dropwise under ice bath Sour (5ml) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later.LCMS detects fully reacting.By reaction solution It is placed in revolving and is concentrated to dryness, then with being dried in vacuo, obtain sepia solid, 920mg, purity: 94%, yield: 100%.Gained Residue is directly used in react in next step.LC-MS(ESI):m/z 330(M+H)⁺。

3, (R)-N- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)propan-2-yl)-5-chlorothiophene-2-carboxamide

Step 2 products therefrom (900mg, 2.73mmol) is dissolved in n,N-Dimethylformamide (5ml) and tetrahydrofuran In (10ml), it is added N under ice bath, N- diisopropyl ethyl amine (1.4g, 10.92mmol) and 5- chlorothiophene -2- carboxylic acid (442mg, 2.73mmol), 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) are eventually adding (1.25g, 3.28mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later.LCMS detects fully reacting, A small amount of water quenching is added to go out, is extracted with ethyl acetate three times, combined organic phase saturated common salt water washing, anhydrous sodium sulfate is done It is dry, concentration.Gained residue obtains off-white powder through column chromatographic purifying (methylene chloride: methanol=50:1~15:1), 595mg, Purity: 96%, yield: 46%.LC-MS(ESI):m/z 474(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 3.0–3.2 (m,2H),3.6–3.8(m,2H),3.9-4.0(m,2H),4.18(s,2H),4.7-4.8(m,1H),6.85(s,1H),7.20– 7.21 (d, 1H, J=4Hz), 7.30-7.33 (d, 2H, J=8.4Hz), 7.56 (s, 1H), 7.61-7.64 (d, 2H, J= 8.8Hz), 7.79-7.81 (d, 1H, J=4.4Hz), 8.91-8.94 (d, 1H, J=7.6Hz), 10.29 (s, 1H).

Embodiment 7:(R) -5-chloro-N- (3- (1-methyl-1H-imidazol-4-yl) -1-oxo-1- (4- (3- Oxomorpholino) phenylamino) propan-2-yl) thiophene-2-carboxamide (FXa-19) preparation

FXa-14 (400mg, 0.84mmol) is dissolved in dimethyl sulfoxide (2ml), under ice bath be added sodium hydroxide (102mg, 2.53mmol), dimethyl suflfate (106mg, 0.84mmol) is added dropwise, ice bath stirring reacts half an hour, goes to room temperature reaction later Half an hour.LCMS detects fully reacting, and a small amount of water quenching is added and goes out, is extracted with ethyl acetate three times, combined organic phase saturation Brine It, anhydrous sodium sulfate is dry, concentration.Gained residue through column chromatographic purifying (methylene chloride: methanol=50:1~ 15:1), faint yellow solid is obtained, 28mg, purity: 95%, yield: 7%.LC-MS(ESI):m/z 488(M+H)⁺；¹H NMR (400MHz,DMSO)δppm 2.9–3.1(m,2H),3.56(s,3H),3.6–3.8(m,2H),3.9-4.0(m,2H),4.18 (s, 2H), 4.7-4.8 (m, 1H), 6.89 (s, 1H), 7.20-7.21 (d, 1H, J=4Hz), 7.30-7.33 (m, 2H), 7.49 (s, 1H), 7.61-7.64 (d, 2H, J=9.2Hz), 7.76-7.78 (d, 1H, J=4Hz), 8.87-8.89 (d, 1H, J= 7.6Hz),10.26(s,1H)。

Embodiment 8:(R)-N- (3- (1H-indol-2-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -5-chlorothiophene-2-carboxamide (FXa-20) preparation

1, intermediate (R)-tert-butyl-3- (1H-indol-2-yl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenylamino)-propan-2-ylcarbamate

4- (4-aminophenyl) morpholin-3-one (500mg, 2.60mmol) is dissolved in N, N- dimethyl formyl In amine (2ml) and tetrahydrofuran (6ml), N, N- diisopropyl ethyl amine (1.2g, 9.1mmol) and Boc-D- are added under ice bath Tryptophan-OH (792mg, 2.60mmol) is eventually adding 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea Hexafluorophosphoric acid ester (HATU) (1.2g, 3.12mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later. LCMS detects fully reacting, and a small amount of water quenching is added and goes out, is extracted with ethyl acetate three times, combined organic phase is washed with saturated common salt It washs, anhydrous sodium sulfate is dry, concentration.Gained residue obtains class through column chromatographic purifying (methylene chloride: methanol=50:1~15:1) White solid, 820mg, purity: 95%, yield: 66%.LC-MS(ESI):m/z479(M+H)⁺。

2, intermediate (R) -2-amino-3- (1H-indol-2-yl)-N- (4- (3-oxomorpholino) is prepared phenyl)propanamide

Step 1 products therefrom (300mg, 0.63mmol) is dissolved in methylene chloride (2ml), trifluoroacetic acid is added dropwise under ice bath (1ml) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later.LCMS detects fully reacting.Reaction solution is set In being concentrated to dryness in revolving, then with vacuum drying, sepia solid is obtained, 237mg, purity: 92%, yield: 100%.Gained is residual Excess is directly used in react in next step.LC-MS(ESI):m/z 379(M+H)⁺。

3, (R)-N- (3- (1H-indol-2-yl) -1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)propan-2-yl)-5-chlorothiophene-2-carboxamide

Step 2 products therefrom (237mg, 0.63mmol) is dissolved in n,N-Dimethylformamide (1ml) and tetrahydrofuran In (5ml), it is added N under ice bath, N- diisopropyl ethyl amine (285mg, 2.2mmol) and 5- chlorothiophene -2- carboxylic acid (123mg, 0.75mmol), 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) are eventually adding (286mg, 0.75mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later.LCMS detects fully reacting, A small amount of water quenching is added to go out, is extracted with ethyl acetate three times, combined organic phase saturated common salt water washing, anhydrous sodium sulfate is done It is dry, concentration.Gained residue obtains off-white powder through column chromatographic purifying (methylene chloride: methanol=50:1~15:1), 46mg, Purity: 96%, yield: 18%.LC-MS(ESI):m/z 524(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 3.1-3.3 (m,2H),3.6–3.7(m,2H),3.9–4.0(m,2H),4.19(s,2H),4.8–4.9(m,1H),6.9–7.1(m,2H), 7.1-7.2 (m, 2H), 7.3-7.4 (m, 3H), 7.6-7.7 (d, 2H, J=8.8Hz), 7.73-7.76 (d, 1H, J=7.6Hz), 7.81-7.83 (d, 1H, J=4.4Hz), 8.0-8.3 (m, 1H), 8.87-8.90 (d, 1H, J=8Hz), 10.33 (s, 1H), 10.80(s,1H)。

Embodiment 9:(R) -1- (4-chlorophenyl) -3- (3- (1-methyl-1H-imidazol-5-yl) -1- Oxo-1- (4- (3-oxomorpholino) phenylamino) propan-2-yl) urea (FXa-23) preparation

1, intermediate (R) -1- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)propan-2-yl)-3-(4-chlorophenyl)urea

By (R) -2-amino-3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino) phenyl) Propanamide (230mg, 0.70mmol) is dissolved in methylene chloride (10ml), is added triethylamine (142mg, 1.4mmol), ice The lower dichloromethane solution (10ml) that 1-chloro-4-isocyanatobenzene is added dropwise of bath, is stirred at room temperature reaction 1 hour. LCMS detects fully reacting, and water quenching is added and goes out, and separates organic phase, and anhydrous sodium sulfate is dry, concentration.Gained residue is chromatographed through column It purifies (methylene chloride: methanol=50:1~15:1), obtains off-white powder, 232mg, purity: 95%, yield: 68%.LC-MS (ESI):m/z 483(M+H)⁺；

2, intermediate (R) -1- (4-chlorophenyl) -3- (1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)-3-(1-trityl-1H-imidazol-5-yl)propan-2-yl)urea

Step 1 products therefrom (180mg, 0.37mmol) is dissolved in methylene chloride (5ml), addition triethylamine (94mg, 0.93mmol) with triphenylchloromethane (125mg, 0.45mmol), reaction 1 hour is stirred at room temperature.LCMS detects fully reacting, adds Enter water quenching to go out, separate organic phase, anhydrous sodium sulfate is dry, concentration.Gained residue through column chromatographic purifying (methylene chloride: methanol= 50:1~15:1), obtain off-white powder, 252mg, purity: 95%, yield: 93%.LC-MS(ESI):m/z 725(M+H)⁺；

3, (R) -1- (4-chlorophenyl) -3- (3- (1-methyl-1H-imidazol-5-yl) -1-oxo- is prepared 1-(4-(3-oxomorpholino)phenylamino)propan-2-yl)urea

Step 2 products therefrom (240mg, 0.32mmol) is dissolved in methylene chloride (2ml), trifluoroacetic acid is added dropwise under ice bath Reaction 2 hours is stirred at room temperature in (1ml).LCMS detects fully reacting, and evaporating solvent under reduced pressure and excessive trifluoroacetic acid, gained are residual Excess obtains off-white powder through column chromatographic purifying (DCM:MeOH=50:1~15:1), 42mg, purity: 96%, yield: 26%. LC-MS(ESI):m/z497(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 3.0–3.2(m,2H),3.69(m,2H),3.83 (s, 3H), 3.96 (m, 2H), 4.1-4.3 (m, 3H), 4.7-4.8 (m, 1H), 6.87-6.89 (d, 1H, J=8.4Hz), 7.2- 7.5(m,7H),7.6(m,2H),8.81(s,1H),9.06(s,1H),10.42(s,1H)。

Embodiment 10:(R) -5-chloro-N- (3- (1-ethyl-1H-imidazol-4-yl) -1-oxo-1- (4- (3- Oxomorpholino) phenylamino) propan-2-yl) thiophene-2-carboxamide (FXa-24) preparation

FXa-14 (200mg, 0.42mmol) is dissolved in n,N-Dimethylformamide (1ml), potassium carbonate is added under ice bath (146mg, 1.06mmol) and bromoethane (55mg, 0.51mmol) reacts at room temperature 3 hours.LCMS detects fully reacting, is added few Amount water quenching is gone out, and is extracted with ethyl acetate three times, combined organic phase saturated common salt water washing, and anhydrous sodium sulfate is dry, concentration. Gained residue obtains off-white powder through column chromatographic purifying (methylene chloride: methanol=50:1~15:1), 32mg, purity: 94%, yield: 15%.LC-MS(ESI):m/z 502(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 2.9(m,2H),3.6– 3.9(t,2H),3.8–4.0(m,4H),4.18(s,2H),4.7-4.8(m,1H),6.96(s,1H),7.20–7.21(d,1H,J =4Hz), 7.29-7.32 (m, 2H), 7.56 (s, 1H), 7.59-7.60 (d, 2H, J=2Hz), 7.61-7.77 (m, 1H), 8.86-8.88 (d, 1H, J=8Hz), 10.24 (s, 1H).

Embodiment 11:(R) -2- (3- (4-fluorophenyl) ureido) -3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino)phenyl)propanamide(FXa-43)

1, intermediate (R) -3- (1- (tert-butoxycarbonyl) -1H-imidazol-5-yl) -2- is prepared ((tert-butoxycarbonyl)-amino)propanoic acid

In tri- mouthfuls of burning product of 50ml, 10g (64.52mmol) histidine is dissolved in 15ml methanol at RT., is then being stirred 1ml water is added under the conditions of mixing, dispersed is presented in system, and 19.55g (193.55mmol, 3eq) triethylamine is added, solution is cooled down To at 0 DEG C, 35.16g (161.29mmol, 2.5eq) (Boc) is slowly added dropwise₂Then the methanol solution of O is stirred at 25 DEG C to molten There is no the histidine solids (about 1.5h) to suspend in liquid, stop reaction, and methanol is removed in rotation under reduced pressure, then use 100ml bis- Chloromethanes dissolution, is added the dry 0.5h of anhydrous magnesium sulfate, filtering, and filtrate is concentrated to get colorless oil and is directly used in next step. [LCMS(m/z):356(M+H)⁺]

2, intermediate (R)-tert-butyl5- (2- ((tert-butoxycarbonyl) amino) -3-oxo-3- is prepared ((4-(3-oxomorpholino)phenyl)amino)propyl)-1H-imidazole-1-carboxylate

In 50ml dry three-necked flask, at room temperature by obtained (R) -3- (1- (tert- of the first step butoxycarbonyl)-1H-imidazol-5-yl)-2-((tert-butoxycarbonyl)amino)propanoic Acid is dissolved in 20ml DMF, and 33.29g (4.0eq) DIEA and 13.39g (1.0eq) 4- (4- is then added under agitation Aminophenyl) -3- morpholone, system is then cooled to 0 DEG C, 29.42g (1.2eq) HATU is added portionwise, later at 25 DEG C It is stirred to react 1h.Stop reaction, 50ml water quenching is added and goes out, is repeatedly extracted with ethyl acetate, saturated common salt water washing organic phase, nothing Aqueous sodium persulfate is dry, is concentrated under reduced pressure, and crude product obtains beige solid product by column chromatographic purifying (MeOH:DCM=1:150) 18.6g (two step yields: 54.5%).[LCMS(m/z):530(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 8.1(1H), 7.2~7.6 (5H), 7.0 (2H), 4.4 (1H), 4.2 (2H), 4.0 (2H), 3.7 (2H), 2.7 (2H), 1.5 (9H), 1.3 (9H).

3, intermediate (R) -2-amino-3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino) is prepared phenyl)propanamide

In 5ml dry flask, at room temperature by (R)-tert-butyl5- (2- ((tert- obtained by upper step butoxycarbonyl)amino)-3-oxo-3-((4-(3-oxomorpholino)phenyl)amino)propyl)-1H- Imidazole-1-carboxylate (150mg) is dissolved in the dry methylene chloride of 1ml, 10 DEG C or so addition 1ml trifluoro second Acid, RT. stir 1h under confined conditions.10 DEG C are cooled to hereinafter, 4ml DIEA is slowly added dropwise, stirring 2h is directly used in next step. [LCMS(m/z):330(M+H)⁺]。

4, (R) -2- (3- (4-fluorophenyl) ureido) -3- (1H-imidazol-5-yl)-N- (4- (3- is prepared oxomorpholino)phenyl)propanamide

FXa-43-A3 reaction system is cooled to 0~5 DEG C, controls in this temperature, is slowly added to FXa-43-A4 system, RT. it being stirred overnight, next day adds water quenching reaction, and organic solvent is removed in rotation, and water phase is washed 3 times with ether, then is extracted repeatedly with DCM, Saturated common salt water washing organic phase, anhydrous magnesium sulfate is dry, is concentrated under reduced pressure, and recrystallizes filtering drying with EA to get product is arrived 13.2mg。[LCMS(m/z):467(M+H)⁺]；¹HNMR (DMSO, 400MHz): δ ppm 10.2 (1H), 8.9 (1H), 7.0~7.5 (9H),6.9(1H),6.5(1H),4.6(1H),4.2(2H),3.9(4H),2.9(2H)。

Embodiment 12:(R) -2- (3- (4-chlorophenyl) ureido) -3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino)phenyl)propanamide(FXa-46)

1, intermediate (R) -3- (1- (tert-butoxycarbonyl) -1H-imidazol-5-yl) -2- is prepared ((tert-butoxycarbonyl)amino)propanoic acid

In 50ml dry three-necked flask, at room temperature by obtained (R) -3- (1- (tert- of the first step butoxycarbonyl)-1H-imidazol-5-yl)-2-((tert-butoxycarbonyl)amino)propanoic Acid is dissolved in 20ml DMF, and 33.29g (4.0eq) DIEA and 13.39g (1.0eq) 4- (4- is then added under agitation Aminophenyl) -3- morpholone, system is then cooled to 0 DEG C, 29.42g (1.2eq) HATU is added portionwise, later at 25 DEG C It is stirred to react 1h.Stop reaction, 50ml water quenching is added and goes out, is repeatedly extracted with ethyl acetate, saturated common salt water washing organic phase, nothing Aqueous sodium persulfate is dry, is concentrated under reduced pressure, and crude product obtains beige solid by column chromatographic purifying (MeOH:DCM=1:150~1:60) Product (two step yields: 54.5%).[LCMS(m/z):530(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm8.1(1H),7.2 ~7.6 (5H), 7.0 (2H), 4.4 (1H), 4.2 (2H), 4.0 (2H), 3.7 (2H), 2.7 (2H), 1.5 (9H), 1.3 (9H)

In 5ml dry flask, at room temperature by second step obtained 150mg (R)-tert-butyl5- (2- ((tert-butoxycarbonyl)amino)-3-oxo-3-((4-(3-oxomorpholino)phenyl)amino) Propyl) -1H-imidazole-1-carboxylate is dissolved in the dry methylene chloride of 1ml, 10 DEG C or so addition 1ml trifluoros Acetic acid, RT. stir 1h under confined conditions.10 DEG C are cooled to hereinafter, 4ml DIEA is slowly added dropwise, stirring 2h is directly used in next Step.[LCMS(m/z):330(M+H)⁺]。

4, (R) -2- (3- (4-chlorophenyl) ureido) -3- (1H-imidazol-5-yl)-N- (4- (3- is prepared oxomorpholino)phenyl)propanamide

FXa-46-A3 reaction system is cooled to 0~5 DEG C, FXa-46-A4 system is slowly added to, is stirred overnight at room temperature, add Organic solvent is removed in water quenching reaction, rotation, and water phase is washed 3 times with ether, then is extracted repeatedly with DCM, and saturated common salt water washing is organic Phase, anhydrous magnesium sulfate are dry, are concentrated under reduced pressure, and are recrystallized and are filtered with EA, and filtration cakes torrefaction is to get arriving product 36.6mg.[LCMS(m/ z):483(M+H)⁺]；¹HNMR (DMSO, 400MHz): δ ppm:12.1 (1H), 10.2 (1H), 8.9 (1H), 7.2~7.6 (9H), 6.9(1H),6.6(1H),4.8(1H),4.2(2H),3.9(2H),3.5(2H)；2.9(2H).

Embodiment 13:(R) -1- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -3- (4-bromophenyl) urea (FXa-49) preparation

1, intermediate (R)-tert-butyl-3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenyl-amino)propan-2-ylcarbamate

3, intermediate (R) -1- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)propan-2-yl)-3-(4-bromophenyl)urea

Step 2 products therefrom (230mg, 0.70mmol) is dissolved in methylene chloride (10ml), addition triethylamine (142mg, 1.4mmol), reaction 1 hour is stirred at room temperature in three resulting dichloromethane solution (10ml) of a dropping step under ice bath.LCMS detection Fully reacting is added water quenching and goes out, and separates organic phase, and anhydrous sodium sulfate is dry, concentration.Gained residue is through column chromatographic purifying (two Chloromethanes: methanol=50:1~15:1), obtain off-white powder, 240mg, purity: 95%, yield: 65%.LC-MS(ESI):m/ z 527(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 2.94(m,2H),3.68(m,2H),4.17(m,2H),4.33(m, 2H),4.60(m,1H),6.55(m,1H),6.86(m,1H),7.2–7.6(m,10H),8.96(s,1H),10.18(s,1H), 11.80(s,1H)。

Embodiment 14:(R) -1- (4-bromophenyl) -3- (3- (1-methyl-1H-imidazol-5-yl) -1- Oxo-1- (4- (3-oxomorpholino) phenylamino) propan-2-yl) urea (FXa-56) preparation

1, intermediate (R) -1- (4-bromophenyl) -3- (1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)-3-(1-trityl-1H-imidazol-5-yl)propan-2-yl)urea

By (R) -1- (3- (1H-imidazol-5-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino)-propan-2-yl) -3- (4-bromophenyl) urea (200mg, 0.40mmol) is dissolved in methylene chloride In (5ml), triethylamine (101mg, 1mmol) and triphenylchloromethane (127mg, 0.46mmol) is added, it is small that reaction 1 is stirred at room temperature When.LCMS detects fully reacting, and water quenching is added and goes out, and separates organic phase, and anhydrous sodium sulfate is dry, concentration.Gained residue is through column Chromatographic purifying (methylene chloride: methanol=50:1~15:1), obtains off-white powder, 257mg, purity: 92%, yield: 88%. LC-MS(ESI):m/z 769(M+H)⁺；

2, intermediate (R) -1- (4-bromophenyl) -3- (1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)-3-(1-methyl-1H-imidazol-5-yl)propan-2-yl)urea

Step 1 products therefrom (250mg, 0.32mmol) is dissolved in n,N-Dimethylformamide (2ml), bicarbonate is added Iodomethane (55mg, 0.39mmol) is added dropwise under ice bath in sodium (67mg, 0.80mmol), goes to reaction 2 hours is stirred at room temperature later. LCMS detects fully reacting, and water quenching is added and goes out, a large amount of solids are precipitated, filters, and filter cake is washed with water, dry, obtains off-white powder, 209mg, purity: 94%, yield: 82%.LC-MS(ESI):m/z 784(M+H)⁺；

3, (R) -1- (4-bromophenyl) -3- (3- (1-methyl-1H-imidazol-5-yl) -1-oxo-1- is prepared (4-(3-oxomorpholino)phenylamino)propan-2-yl)urea

Step 2 products therefrom (180mg, 0.23mmol) is dissolved in methylene chloride (2ml), trifluoroacetic acid is added dropwise under ice bath Reaction 2 hours is stirred at room temperature in (1ml).LCMS detects fully reacting, and evaporating solvent under reduced pressure and excessive trifluoroacetic acid, gained are residual Excess obtains off-white powder through column chromatographic purifying (DCM:MeOH=50:1~15:1), 46mg, purity: 96%, yield: 37%. LC-MS(ESI):m/z541(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 3.0–3.2(m,2H),3.6(m,2H),3.8(s, 3H), 3.96 (m, 2H), 4.18 (m, 2H), 4.71-4.73 (m, 1H), 6.94-6.96 (d, 1H, J=8.4Hz), 7.3-7.5 (m, 8H),7.6(m,2H),8.81(s,1H),9.14(s,1H),10.44(s,1H).

Embodiment 15:(R) -5-chloro-N- (3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- Oxomorpholino) phenylamino) propan-2-yl) thiophene-2-carboxamide (FXa-60) preparation

1, intermediate (R)-tert-butyl-3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenylamino)propan-2-yl-carbamate

By 4- (4-aminophenyl) morpholin-3-one (1.37g, 7.11mmol) and Boc-D-Tryptophan- OH (2.0g, 7.11mmol) is dissolved in n,N-Dimethylformamide (6ml) and tetrahydrofuran (20ml), and N, N- bis- are added under ice bath Diisopropylethylamine (3.22g, 24.89mmol) is eventually adding 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea Hexafluorophosphoric acid ester (HATU) (3.24g, 8.53mmol) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later. LCMS detects fully reacting, reaction solution is concentrated under reduced pressure, reaction solution is slowly added to about by most of tetrahydrofuran of going out It is stirred in 100ml water, a large amount of off-white powders is precipitated, filtered, drying is concentrated under reduced pressure, obtains off-white powder, 1.5g, purity: 92%, yield: 47%.LC-MS(ESI):m/z 456(M+H)⁺。

2, intermediate (R) -2-amino-3- (4-hydroxyphenyl)-N- (4- (3-oxomorpholino) is prepared phenyl)-propanamide

Step 1 products therefrom (300mg, 0.66mmol) is dissolved in methylene chloride (2.0ml), trifluoro second is added dropwise under ice bath Sour (1.0ml) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later.LCMS detects fully reacting.It will reaction Liquid is placed in revolving and is concentrated to dryness, then with being dried in vacuo, obtains brown solid, 230mg, purity: 97%, yield: 98%.Gained is residual Excess is directly used in react in next step.LC-MS(ESI):m/z 356(M+H)⁺。

3, (R) -5-chloro-N- (3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)-phenylamino)propan-2-yl)thiophene-2-carboxamide

Step 2 products therefrom (312mg, 0.88mmol) is dissolved in n,N-Dimethylformamide (1ml) and tetrahydrofuran In (5ml) mixed solution, adds stirring under 5- chlorothiophene -2- carboxylic acid (143mg, 0.88mmol) ice bath and N, N- diisopropyl is added After stirring 6min, 2- (7- azo benzotriazole)-N, N, N', N'- tetramethyl is added in base ethamine (0.77ml, 4.4mmol) Urea hexafluorophosphoric acid ester (HATU) (403mg, 1.06mmol) goes to and reaction 2 hours is stirred at room temperature.LCMS detects fully reacting, adds Entering a certain amount of water, off-white powder is precipitated, filters, gains are crossed reverse phase column purification, obtain off-white powder by vacuum drying, 90mg, purity: 98%, yield: 28%.LC-MS(ESI):m/z500(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 2.9– 3.01(m,2H),3.59–3.69(m,4H),3.95(m,2H),4.17(s,2H),4.71(s,1H),6.65(m,2H),7.14 (m,3H),7.33(m,2H,),7.61(m,2H),7.8(s,1H),8.81(s,1H),9.20(s,1H).

Embodiment 16:(R) -1- (4-bromophenyl) -3- (3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- Oxomorpholino) phenylamino) propan-2-yl) urea (FXa-61) preparation

Intermediate (R) -2-amino-3- (4-hydroxyphenyl)-N- (4- (3-oxomorpholino) phenyl) - The synthetic method of propanamide is the same as COMPOUNDS EXAMPLE 15.

1,1-bromo-4-isocyanatobenzene is prepared

Para-bromoaniline (170mg, 0.99mmol) is dissolved in methylene chloride (4.0ml), addition triethylamine (200mg, 1.98mmol), methylene chloride (2.0ml) solution of triphosgene (293mg, 0.99mmol) is added dropwise under ice bath, reaction is stirred at room temperature 15 minutes, it is heated to back flow reaction later 1 hour.LCMS detects fully reacting.Acquired solution is directly used in reacts in next step.

2, (R) -1- (4-bromophenyl) -3- (3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenylamino)propan-2-yl)urea

By (R) -2-amino-3- (4-hydroxyphenyl)-N- (4- (3-oxomorpholino) phenyl) - Propanamide (230mg, 0.65mmol) is dissolved in methylene chloride (10ml), is added triethylamine (130mg, 1.3mmol), ice The dichloromethane solution (10ml) for bathing lower one products therefrom of a dropping step, is stirred at room temperature reaction 1 hour.LCMS detection has been reacted Entirely, water quenching is added to go out, separates organic phase, anhydrous sodium sulfate is dry, concentration.Gained residue through column chromatographic purifying (methylene chloride: Methanol=50:1~15:1), obtain off-white powder, 100mg, purity: 98%, yield: 28%.LC-MS(ESI):m/z 554 (M+H)⁺；¹H NMR(400MHz,DMSO)δppm 2.8-3.0(m,2H),3.69(m,2H),3.95(m,2H),4.18(m,2H), 4.55 (m, 1H), 6.45-6.47 (d, 1H, J=8Hz), 6.64-6.66 (d, 2H, J=8.2Hz), 7.0 (m, 2H), 7.3-7.4 (m,6H,),7.5(m,2H),8.87(s,1H),9.20(s,1H),10.20(s,1H)。

Embodiment 17:(R) -5-chloro-N- (3- (4-methoxyphenyl) -1-oxo-1- (4- (3- Oxomorpholino) phenylamino) propan-2-yl) thiophene-2-carboxamide (FXa-62) preparation

1, intermediate (R)-methyl 2- (tert-butoxycarbonyl) -3- (4-methoxyphenyl) is prepared propanoate

Boc-D-Tryptophan-OH (500mg, 1.78mmol) is dissolved in n,N-Dimethylformamide (4.0ml), ice It is stirred under bath wait be completely dissolved, potassium carbonate (700mg, 3.0mmol) and iodomethane (550mg, 2.20mmol) is once added later, Room temperature reaction 3 hours is gone to later.TLC detects raw material fully reacting, reaction solution is carried out extraction processing, organic phase is concentrated under reduced pressure It is dry, obtain brown oil liquid, 500mg, purity: 95%, yield: 90%.¹H NMR(400MHz,CDCl3)δppm 1.31 (s,9H),2.78–2.85(m,2H),3.65(s,3H),4.03-4.23(s,3H),5.37(s,1H),6.62-7.03(m,4H).

2, intermediate (R) -2- (tert-butoxycarbonyl) -3- (4-methoxyphenyl) propanoic is prepared acid

Step 1 products therefrom (500mg, 1.61mmol) is dissolved in anhydrous methanol (3.0ml), is slowly added under ice bath 10% sodium hydrate aqueous solution (1.5ml) is stirred to react 1.5 hours.TLC detects raw material fully reacting, water-soluble with the hydrochloric acid of 1N Reaction solution PH is adjusted to 6 left back, extraction processings by liquid, and drying is concentrated under reduced pressure in organic phase, obtains brown solid, 350mg, purity: 95%, Yield: 75%.¹H NMR(400MHz,DMSO)δppm 1.33(s,9H),2.79–2.88(m,2H),3.60-3.71(s,3H), 6.84-7.21(m,4H).

3, intermediate (R)-tert-butyl-3- (4-methoxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)-phenylamino)propan-2-ylcarbamate

By step 2 products therefrom (350mg, 1.185mmol) and 4- (4-aminophenyl) morpholin-3-one (230mg, 1.185mmol) is dissolved in n,N-Dimethylformamide (1ml) and tetrahydrofuran (5ml), and N, N- bis- are added under ice bath Diisopropylethylamine (536mg, 4.147mmol) finally plus (540mg, 1.422mmol) is stirred to react 15 minutes under ice bath, it After go to room temperature reaction 3 hours.LCMS detects fully reacting, reaction solution is concentrated under reduced pressure, most of tetrahydrofuran of going out, Reaction solution is slowly added to stir in about 80ml water, a large amount of off-white powders are precipitated, is filtered, drying is concentrated under reduced pressure, obtains off-white color Solid, 390mg, purity: 99%, yield: 71%.LC-MS(ESI):m/z 470(M+H)⁺。

4, intermediate (R) -2-amino-3- (4-methoxyphenyl)-N- (4- (3-oxomorpholino) is prepared phenyl)-propanamide

Step 3 products therefrom (200mg, 0.427mmol) is dissolved in methylene chloride (2.0ml), trifluoro is added dropwise under ice bath Acetic acid (1.0ml) is stirred to react 15 minutes under ice bath, goes to room temperature reaction 3 hours later.LCMS detects fully reacting.It will be anti- It answers liquid to be placed in revolving to be concentrated to dryness, then with being dried in vacuo, gained residue is directly used in react in next step.LC-MS(ESI): m/z 370(M+H)⁺。

5, (R) -5-chloro-N- (3- (4-methoxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)-phenylamino)propan-2-yl)thiophene-2-carboxamide

Step 4 products therefrom (312mg, 0.362mmol) and 7 (58.9mg, 0.362mmol) are dissolved in N, N- dimethyl methyl In amide and tetrahydrofuran mixed solution, n,N-diisopropylethylamine (0.32ml, 1.81mmol) is added in stirring under ice bath, stirring After 6min, addition 2- (7- azo benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HATU) (165mg, 0.434mmol), it goes to and reaction 2 hours is stirred at room temperature.LCMS detects fully reacting, and a certain amount of water is added, and it is solid that off-white color is precipitated Body filters, and gains are crossed reverse phase column purification, obtain off-white powder by vacuum drying, 40mg, purity: 98%, yield: 22%. LC-MS(ESI):m/z 514(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 2.97–3.33(m,4H),3.69(s,5H), 3.95 (m, 2H), 4.17 (s, 2H), 6.83 (m, 2H), 7.19-7.34 (m, 5H), 7.62 (d, 2H, J=6Hz), 7.81 (d, 1H, ), J=4Hz 8.92 (d, 1H, J=6Hz)

Embodiment 18:(R) -1- (3- (1H-indol-2-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -3- (3-bromophenyl) urea (FXa-63) preparation

1,1-bromo-3-isocyanatobenzene is prepared

M-bromoaniline (110mg, 0.64mmol) is dissolved in methylene chloride (3.0ml), addition triethylamine (130mg, 1.28mmol), methylene chloride (5.0ml) solution of triphosgene (190mg, 0.64mmol) is added dropwise under ice bath, reaction is stirred at room temperature 15 minutes, it is heated to back flow reaction later 1 hour.LCMS detects fully reacting.Acquired solution is directly used in reacts in next step.

2, (R) -1- (3- (1H-indol-2-yl) -1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)-propan-2-yl)-3-(3-bromophenyl)urea

By (R) -2-amino-3- (1H-indol-2-yl)-N- (4- (3-oxomorpholino) phenyl) Propanamide (156mg, 0.42mmol) is dissolved in methylene chloride (10ml), is added triethylamine (0.12ml, 0.84mmol), Reaction 1 hour is stirred at room temperature in one resulting dichloromethane solution (8ml) of a dropping step under ice bath.LCMS detects fully reacting, adds Enter water quenching to go out, separate organic phase, anhydrous sodium sulfate is dry, concentration.Gained residue is through column chromatographic purifying (methylene chloride: methanol =50:1~15:1), obtain off-white powder, 90mg, purity: 98%, yield: 38%.LC-MS(ESI):m/z 568(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 3.17(m,2H),3.95(m,2H),4.08(m,2H),4.8-5.0(m,1H),6.5(m, 1H),6.85(m,2H),7.13(m,2H),7.3-7.4(m,6H),7.4(m,1H),7.57(m,2H),8.87(s,1H),10.24 (s,1H)。

Embodiment 19:(R) -1- (4-chlorophenyl) -3- (3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- Oxomorpholino) phenylamino) propan-2-yl) urea (FXa-65) preparation

1,1-chloro-4-isocyanatobenzene is prepared

Parachloroanilinum (85mg, 0.66mmol) is dissolved in methylene chloride (3.0ml), addition triethylamine (135mg, 1.3mmol), methylene chloride (2.0ml) solution of triphosgene (196mg, 0.66mmol) is added dropwise under ice bath, reaction 15 is stirred at room temperature Minute, it is heated to back flow reaction later 1 hour.LCMS detects fully reacting.Acquired solution is directly used in reacts in next step.

2, (R) -1- (4-chlorophenyl) -3- (3- (4-hydroxyphenyl) -1-oxo-1- (4- (3- is prepared oxomorpholino)phenylamino)propan-2-yl)urea

By (R) -2-amino-3- (4-hydroxyphenyl)-N- (4- (3-oxomorpholino) phenyl) - Propanamide (150mg, 0.42mmol) is dissolved in methylene chloride (10ml), is added triethylamine (0.12ml, 0.84mmol), Reaction 1 hour is stirred at room temperature in one resulting dichloromethane solution (5ml) of a dropping step under ice bath.LCMS detects fully reacting, adds Enter water quenching to go out, separate organic phase, anhydrous sodium sulfate is dry, concentration.Gained residue is through column chromatographic purifying (methylene chloride: methanol =50:1~15:1), obtain off-white powder, 72mg, purity: 98%, yield: 34%.LC-MS(ESI):m/z 509(M+H)⁺；¹H NMR(400MHz,DMSO)δppm 2.82(m,1H),2.94(m,1H),3.69(m,2H),3.96(m,2H),4.18(s, 2H), 4.556-4.570 (m, 1H, J=1.6Hz), 6.45 (m, 1H), 6.646-6.668 (m, 2H, J=8.8Hz), 7.25- 7.37(m,6H),7.57(m,2H),8.87(s,1H),9.20(s,1H),10.24(s,1H).

Embodiment 20:(R) -1- (3- (1H-indol-2-yl) -1-oxo-1- (4- (3-oxomorpholino) Phenylamino) propan-2-yl) -3- (4-chlorophenyl) urea (FXa-66) preparation

1,1-chloro-4-isocyanatobenzene is prepared

Parachloroanilinum (650mg, 5.1mmol) is dissolved in methylene chloride (15ml), addition triethylamine (1.03g, 10.2mmol), methylene chloride (5.0ml) solution of triphosgene (1.51g, 5.1mmol) is added dropwise under ice bath, reaction 15 is stirred at room temperature Minute, it is heated to back flow reaction later 1 hour.LCMS detects fully reacting.Acquired solution is directly used in reacts in next step.

2, (R) -1- (3- (1H-indol-2-yl) -1-oxo-1- (4- (3-oxomorpholino) is prepared phenylamino)-propan-2-yl)-3-(4-chlorophenyl)urea

By (R) -2-amino-3- (1H-indol-2-yl)-N- (4- (3-oxomorpholino) phenyl) Propanamide (1.0g, 2.64mmol) is dissolved in methylene chloride (20ml), is added triethylamine (270mg, 2.64mmol), ice Lower one resulting dichloromethane solution (20ml) of a dropping step is bathed, reaction 1 hour is stirred at room temperature.LCMS detects fully reacting, adds Enter water quenching to go out, separate organic phase, anhydrous sodium sulfate is dry, concentration.Gained residue is through column chromatographic purifying (methylene chloride: methanol =50:1~15:1), obtain off-white powder, 560mg, purity: 96%, yield: 40%.LC-MS(ESI):m/z 532(M+H)⁺；¹H NMR (400MHz, DMSO) δ ppm 2.5 (s, 1H), 3.2-3.3 (m, 2H), 3.7 (d, 2H), 3.9 (s, 2H), 4.2 (s, 2H), 4.6-4.7(m,1H),6.4(s,1H),6.9(m,1H),7.04(m,1H),7.12-7.13(m,1H),7.26-7.32(m,7H), 7.6(m,3H),8.86(s,1H),10.2(s,1H),10.85(s,1H)。

Embodiment 21:(S) -2- (3- (4-bromophenyl) ureido) -3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino) phenyl) propanamide (FXa-80) preparation

1, intermediate (S)-tert-butyl (3- (1H-imidazol-5-yl) -1-oxo-1- ((4- (3- is prepared oxomorpholino)phenyl)amino)propan-2-yl)carbamate

In 50ml dry three-necked flask, 2g N-Boc-L- histidine is dissolved in 15ml DMF, then in stirring bar 4.03g (4.0eq) DIEA and 1.51g (1.0eq) 4- (4- aminophenyl) -3- morpholone is added under part, then system cools down To 0 DEG C, 3.58g (1.2eq) HATU is added portionwise, is stirred to react 70h at 25 DEG C later.Stop reaction, 60ml water quenching is added It goes out, is repeatedly extracted with ethyl acetate, saturated common salt water washing organic phase, anhydrous sodium sulfate is dry, is concentrated under reduced pressure, and crude product passes through column Chromatographic purifying (MeOH:DCM=1:100~1:10) obtains beige solid product 2.2g, and [LCMS (m/z): 430 (M+H)⁺]；¹HNMR (DMSO, 400MHz): δ ppm 8.1 (1H), 7.2~7.6 (5H), 7.0 (2H), 4.4 (1H), 4.2 (2H), 4.0 (2H), 3.7(2H),2.7(2H),1.3(9H).

2, intermediate (S) -2-amino-3- (1H-imidazol-5-yl)-N- (4- (3-oxomorpholino) is prepared phenyl)propanamide

In 5ml dry flask, at room temperature by step 1 obtained 245mg (S)-tert-butyl (3- (1H- imidazol-5-yl)-1-oxo-1-((4-(3-oxomorpholino)phenyl)amino)propan-2-yl) Carbamate is dissolved in the dry methylene chloride of 2ml, and 10 DEG C or so addition 1ml trifluoroacetic acids, RT. stirs 2h under confined conditions. It is spin-dried for reaction system, obtains the trifluoroacetate of FXa-80-A2, is directly used in next step.[LCMS(m/z):330(M+H)⁺].

3, (S) -2- (3- (4-bromophenyl) ureido) -3- (1H-imidazol-5-yl)-N- (4- (3- is prepared oxomorpholino)phenyl)propanamide

FXa-80-A2 trifluoroacetate is dissolved in DCM, is cooled to 0~5 DEG C, controls in this temperature, 2.5g is slowly added dropwise DIEA, RT. are stirred to react 1h, are cooled to 0 DEG C or so, are slowly added to FXa-80-A3 system, and RT. stirs 2h, water quenching is added to go out instead It answers, organic solvent is removed in rotation, and water phase is washed 3 times with ether, then is extracted repeatedly with DCM, saturated common salt water washing organic phase, anhydrous sulphur Sour magnesium is dry, is concentrated under reduced pressure, and is recrystallized and is filtered with EA, and filtration cakes torrefaction is to get arriving product 18.7mg.[LCMS(m/z):527(M+ H)⁺]；¹HNMR(DMSO,400MHz):δppm 10.2(1H),9.0(1H),7.7(1H),7.6(2H)7.3(6H),6.9(1H), 6.6(1H),4.6(1H),4.2(2H),3.9(4H),3.1(2H).

Embodiment 22:(S) -2- (5-chlorothiophene-2-carboxamido)-N1- (4- (3- Oxomorpholino) phenyl) succinamide (FXa-84) preparation

1, intermediate (S) -2-amino-4-methoxy-4-oxobutanoic acid hydrochloride is prepared

In 50ml three-necked flask, 5g (39.59mmol, 1.0eq) D-ASP is dissolved in 30ml methanol at RT. In, it is cooled at 0 DEG C, 4.43g (39.6mmol, 1.0eq) SOCl is slowly added dropwise₂, it is warmed to room temperature naturally and is stirred to react 2h.To anti- Addition 60ml ether in system is answered, decompression filters, and filter cake is washed with ether, dries filter cake and obtain product 4.73g, [LCMS (m/ z):148(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 8.5(2H),4.2(1H),3.6(3H),2.9(2H。

2, intermediate (S) -2- ((tert-butoxycarbonyl) amino) -4-methoxy-4- is prepared oxobutanoic acid

In 100ml dry three-necked flask, at room temperature by 4.73g (25.7mmol, 1.0eq) (S) -2-amino-4- Methoxy-4-oxobutanoic acid hydrochloride is dissolved in the THF/H of 50ml 1:1₂5.45g is added in O system Na₂CO₃(51mmol, 2.0eq), is stirred at room temperature 0.5h, is cooled to 5 DEG C or so, and 6.17g (51.2mmol, 2.0eq) is slowly added dropwise (Boc)₂The THF solution of O, RT are stirred overnight.THF is removed in rotation, three times with EA aqueous phase extracted, is adjusted water phase PH=4 or so, is spin-dried for water White solid is mutually obtained, with more washing solids of EA to not until having product signal.Rotation goes EA to obtain product 2.54g.[LCMS (m/z):248(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 6.5(1H),4.2(1H),3.6(3H),2.7(1H),2.6 (1H),1.4(9H)。

3, intermediate (S)-methyl-3- ((tert-butoxycarbonyl) amino) -4-oxo-4- ((4- (3- is prepared oxomorpholino)phenyl)amino)butanoate

In 50ml dry three-necked flask, 1.94g (7.8mmol, 1.0eq) (S) -2- ((tert- at room temperature Butoxycarbonyl) amino) -4-methoxy-4-oxobutanoic acid is dissolved in 12ml DMF, then in stirring bar 3.04g (23.56mmol, 3.0eq) DIEA and 1.5g (7.8mmol, 1.0eq) 4- (4- aminophenyl) -3- morpholine is added under part Then system is cooled to 0 DEG C by ketone, 3.58g (9.43mmol, 1.2eq) HATU is added portionwise, is stirred to react at 25 DEG C later Overnight.Stop reaction, 50ml water quenching is added and goes out, is repeatedly extracted with ethyl acetate, saturated common salt water washing organic phase, anhydrous slufuric acid Sodium is dry, is concentrated under reduced pressure, and crude product obtains product as light yellow solid by column chromatographic purifying (MeOH:DCM=1:150~1:60) 2.6g。[LCMS(m/z):422(M+H)⁺]；¹HNMR(CDCl₃,400MHz):δppm8.7(1H)7.6(2H),7.3(2H),5.9 (1H),4.7(1H),4.4(2H),4.1(2H),3.7(5H),3.0(1H),2.8(1H)1.5(9H).

4, intermediate (S)-methyl-3-amino-4-oxo-4- ((4- (3-oxomorpholino) phenyl) is prepared amino)butanoate

In 50ml dry flask, 1g (2.37mmol, 1.0eq) (S)-methyl 3- ((tert- at room temperature Butoxycarbonyl) amino) -4-oxo-4- ((4- (3-oxomorpholino) phenyl) amino) butanoate is dissolved in In 5ml dry methylene chloride, 10 DEG C or so addition 1.8ml trifluoroacetic acids, RT. is stirred overnight under confined conditions.Closed preservation It is directly used in next step.[LCMS(m/z):322(M+H)⁺]。

5, intermediate (S)-methyl-3- (5-chlorothiophene-2-carboxamido) -4-oxo-4- is prepared ((4-(3-oxomorpholino)phenyl)amino)butanoate

FXa-84-A4 reaction system is cooled to 0~5 DEG C, 5.07g DIEA is slowly added dropwise, reaction 0.5h is stirred at room temperature, 385mg 5- chlor-2-thiophenecar-boxylic acid is added, 10 DEG C or so, 1.08g HATU is added portionwise, RT. is stirred to react 3h, and add water quenching to go out, It is extracted with EA more times, saturated common salt water washing, anhydrous sodium sulfate is dry, and the crude product of concentration, column chromatographic purifying obtains product 972mg。

[LCMS(m/z):466(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 10.3(1H),9.0(1H),7.8 (1H),7.6(2H),7.3(2H),7.1(1H),4.9(1H),4.2(2H),3.9(2H),3.6(5H),2.9(1H),2.8(1H)。

6, intermediate (S) -3- (5-chlorothiophene-2-carboxamido) -4-oxo-4- ((4- (3- is prepared oxomorpholino)phenyl)amino)butanoic acid

By 900mg (1.93mmol, 1.0eq) (S)-methyl-3- (5-chlorothiophene-2- Carboxamido) -4-oxo-4- ((4- (3-oxomorph olino) phenyl) amino) butanoate is dissolved in 10ml THF/MeOH/H₂243mg (243mg, 3.0eq) LiOH.H is added portionwise at 0 DEG C in O (3:1:1)₂O, RT. are stirred to react 1.5h, rotation Solvent to be removed, is dissolved in water, three times with EA aqueous phase extracted, adjusts water phase PH=3 or so, EA extraction is multiple, saturated common salt water washing, Anhydrous sodium sulfate is dry, and concentration crude product column chromatographic purifying obtains sterling 120mg.

[LCMS(m/z):452(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 10.1(1H),9.0(1H),7.7 (1H),7.6(2H)7.3(2H),7.2(1H),4.7(1H),4.2(2H),3.9(2H),3.6(2H),3.0(1H),2.8(1H)。

7, (S) -2- (5-chlorothiophene-2-carboxamido)-N1- (4- (3-oxomorpholino) is prepared phenyl)Succinamide

In 10ml dry flask, 89mg (0.2mmol, 1.0eq) (S) -3- (5- at room temperature chlorothiophene-2-carboxamido)-4-oxo-4-((4-(3-oxomorpholino)phenyl)amino) Butanoic acid is dissolved in 2ml DMF, and 99mg (0.8mmol, 4.0eq) DIEA, 41mg is then added under agitation (0.8mmol,4.0eq)NH₄Cl, 73.4mg (0.4mmol, 2.0eq) EDCI and 52mg (0.4mmol, 2.0eq), later 25 It is stirred to react at DEG C overnight.Stop reaction, 20ml water quenching is added and goes out, is repeatedly extracted with ethyl acetate, saturated common salt water washing has Machine phase, anhydrous sodium sulfate is dry, is concentrated under reduced pressure, and crude product EA is recrystallized to give product 50mg.

[LCMS(m/z):451(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 10.1(1H),9.0(1H),7.7 (1H),7.6(2H),7.5(1H),7.3(2H),7.2(2),4.7(1H),4.2(2H),3.9(2H),3.6(2H),3.0(1H), 2.8(1H)。

Embodiment 23:(R) -5-chloro-N- (1-oxo-1- ((4- (3-oxomorpholino) phenyl) amino) - 3-phenylpropan-2-yl) the preparation of thiophene-2-carboxamide (FXa-93)

1, intermediate (R) -2- ((tert-butoxycarbonyl) amino) -3-phenylpropanoic acid is prepared

In the dry single-necked flask of 50ml, 0.5g (R) -2-amino-3-phenylpropanoic acid is dissolved in 4ml 0.127g (1.05eq) sodium hydroxide is added in THF and 1ml water, under RT. and does alkali, stirs 30min, is cooled to 0 DEG C or so, slowly It is added dropwise 0.727g (1.1eq) (Boc)₂The THF solution of O rises to RT reaction 3h naturally.Stop reaction, low temperature adjusts PH=3~4 Left and right, with EA extraction extraction 5 times or so, saturated common salt water washing, drying is concentrated to get light brown grease 412mg.

[LCMS(m/z):266(M+H)⁺]。

2, intermediate (R)-tert-butyl (1-oxo-1- ((4- (3-oxomorpholino) phenyl) is prepared amino)-3-phenylpropan-2-yl)carbamate

In the dry single-necked flask of 50ml, 200mg (R) -2- ((tert-butoxycarbonyl) amino) -3- Phenylpropanoic acid is dissolved in 4ml DCM, and 389mg (4.0eq) DIEA and 145mg (1.0eq) are sequentially added under RT. 4- (4-aminophenyl) morpholin-3-one, 10 DEG C or so add 344mg (1.2eq) HATU in batches, and reaction is stirred at room temperature 3h or so.Stop reaction, add water quenching reaction, extracted repeatedly with DCN, with saturated common salt water washing, anhydrous magnesium sulfate is dry, subtracts Pressure concentration, column chromatographic purifying obtain sterling 357mg.

[LCMS(m/z):440(M+H)⁺]。

3, intermediate (R) -2-amino-N- (4- (3-oxomorpholino) phenyl) -3- is prepared phenylpropanamide

In 50ml dry flask, by 300mg ((R)-tert-butyl (1-oxo-1- ((4- (3- under RT. Oxomorpholino) phenyl) amino) -3-phenylpropan-2-yl) carbamate is dissolved in the dry dichloromethane of 4ml In alkane, 0.5ml trifluoroacetic acid is added, enclosed system is stirred to react 1h, it is spare to be spin-dried for solvent.

[LCMS(m/z):340(M+H)⁺]。

4, intermediate (R) -5-chloro-N- (1-oxo-1- ((4- (3-oxomorpholino) phenyl) is prepared amino)-3-phenyl propan-2-yl)thiophene-2-carboxa

At 0 DEG C, 3g DIEA is added in step 3 products therefrom system, RT. is stirred to react 0.5h, is added under RT. 111mg5-chlorothiophene-2-carboxylic acid, 10 DEG C or less addition 312mg HATU, RT are stirred to react 1h, Stop reaction, water quenching is added to go out, extracted with EA more times, saturated common salt water washing, anhydrous sodium sulfate is dry, the sterling of concentrating and purifying 174mg。[LCMS(m/z):484(M+H⁺)]。

¹HNMR(MeOD,400MHz):δppm 7.6(3H),7.3(7H),7.0(1H),4.9(1H),4.3(2H),4.0 (2H),3.8(2H),3.2(1H),3.1(1H)。

Embodiment 24:(R) -4- (5-chlorothiophene-2-carboxamido) -5-oxo-5- ((4- (3- Oxomorpholino) phenyl) amino) pentanoic acid (FXa-98) preparation

1, intermediate (R) -2-amino-4-methoxy-4-oxobutanoic acid hydrochloride is prepared

In 50ml three-necked flask, 5g (39.59mmol, 1.0eq) ASPARTIC ACID is dissolved in 30ml methanol at RT. In, it is cooled at 0 DEG C, 4.43g (39.6mmol, 1.0eq) SOCl is slowly added dropwise₂, it is warmed to room temperature naturally and is stirred to react 2h.To anti- Addition 60ml ether in system is answered, decompression filters, and filter cake is washed with ether, dries filter cake and obtain product 4.73g.

[LCMS(m/z):148(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 8.5(2H),4.2(1H),3.6(3H), 2.9(2H)。

2, intermediate (R) -2- ((tert-butoxycarbonyl) amino) -4-methoxy-4- is prepared oxobutanoic acid

In 100ml dry three-necked flask, at room temperature by 4.73g (25.7mmol, 1.0eq) (R) -2-amino-4- Methoxy-4-oxobutanoic acid hydrochloride is dissolved in the THF/H of 50ml 1:1₂5.45g is added in O system Na₂CO₃(51mmol, 2.0eq), is stirred at room temperature 0.5h, is cooled to 5 DEG C or so, and 6.17g (51.2mmol, 2.0eq) is slowly added dropwise (Boc)₂The THF solution of O, RT are stirred overnight.THF is removed in rotation, three times with EA aqueous phase extracted, is adjusted water phase PH=4 or so, is spin-dried for water White solid is mutually obtained, with more washing solids of EA to not until having product signal.Rotation goes EA to obtain product 2.54g.

[LCMS(m/z):248(M+H)⁺]；¹HNMR(DMSO,400MHz):δppm 6.5(1H),4.2(1H),3.6(3H), 2.7(1H),2.6(1H),1.4(9H)。

3, intermediate (R)-methyl 3- ((tert-butoxycarbonyl) amino) -4-oxo-4- ((4- (3- is prepared oxomorpholino)phenyl)amino)butanoate

In 50ml dry three-necked flask, (R) -2- ((tert-butoxycarbonyl) is added at room temperature Amino) -4-methoxy-4-oxobutanoic acid (1.94g, 7.8mmol, 1.0eq) is dissolved in 12ml DMF, is then existed 3.04g (2356mmol, 3.0eq) DIEA and 1.5g (7.8mmol, 1.0eq) 4- (4- aminophenyl) -3- is added under stirring condition Then system is cooled to 0 DEG C by morpholone, 3.58g (9.43mmol, 1.2eq) HATU is added portionwise, stirs at 25 DEG C later Reaction is overnight.Stop reaction, 50ml water quenching is added and goes out, is repeatedly extracted with ethyl acetate, saturated common salt water washing organic phase is anhydrous Sodium sulphate is dry, is concentrated under reduced pressure, and crude product obtains light yellow solid production by column chromatographic purifying (MeOH:DCM=1:150~1:60) Object 2.6g [LCMS (m/z): 422 (M+H)⁺]。

¹HNMR(CDCl₃,400MHz):δppm 8.7(1H)7.6(2H),7.3(2H),5.9(1H),4.7(1H),4.4 (2H), 4.1 (2H), 3.7 (5H), 3.0 (1H), 2.8 (1H), 1.5 (9H).

4, intermediate (R)-methyl 3-amino-4-oxo-4- ((4- (3-oxomorpholino) phenyl) is prepared amino)butanoate

In 50ml dry flask, (R)-methyl 3- ((tert-butoxycarbonyl) is added at room temperature Amino) -4-oxo-4- ((4- (3-oxomorpholino) phenyl) amino) butanoate (1.00g, 2.37mmol, It 1.0eq) is dissolved in the dry methylene chloride of 5ml, 10 DEG C or so addition 1.8ml trifluoroacetic acids, RT. is stirred under confined conditions Night.Closed preservation is directly used in next step.

[LCMS(m/z):322(M+H)⁺]；

5, intermediate (R)-methyl-3- (5-chlorothiophene-2-carboxamido) -4-oxo-4- is prepared ((4-(3-oxomorpholino)phenyl)amino)butanoate

FXa-94-A4 reaction system is cooled to 0~5 DEG C, 5.07g DIEA is slowly added dropwise, reaction 0.5h is stirred at room temperature, 385mg 5- chlor-2-thiophenecar-boxylic acid is added, 10 DEG C or so, 1.08g HATU is added portionwise, RT. is stirred to react 3h, and add water quenching to go out, It is extracted with EA more times, saturated common salt water washing, anhydrous sodium sulfate is dry, and the crude product of concentration, column chromatographic purifying obtains product 972mg。[LCMS(m/z):466(M+H)⁺]。

¹HNMR(DMSO,400MHz):δppm 10.3(1H),9.0(1H),7.8(1H),7.6(2H),7.3(2H),7.1 (1H),4.9(1H),4.2(2H),3.9(2H),3.6(5H),2.9(1H),2.8(1H)。

6, (R) -3- (5-chlorothiophene-2-carboxamido) -4-oxo-4- ((4- (3- is prepared oxomorpholino)phenyl)amino)butanoic acid

By (R)-methyl-3- (5-chlorothiophene-2-carboxamido) -4-oxo-4- ((4- (3- Oxomorpholino) phenyl) amino) butanoate (900mg, 1.93mmol, 1.0eq) is dissolved in 10ml THF/MeOH/ H₂243mg (243mg, 3.0eq) LiOH.H is added portionwise at 0 DEG C in O (3:1:1)₂O, RT. are stirred to react 1.5h, and rotation is removed solvent, added Water dissolution three times with EA aqueous phase extracted adjusts water phase PH=3 or so, EA extraction is multiple, saturated common salt water washing, anhydrous slufuric acid Sodium is dry, and concentration crude product column chromatographic purifying obtains sterling 120mg.

Embodiment 25:(R) -2- (5-chlorothiophene-2-carboxamido)-N1- (4- (3- Oxomorpholino) phenyl) pentanediamide (FXa-99) preparation

In 10ml dry flask, 24 gained FXa-98 of 89mg embodiment is dissolved in 2ml DMF at room temperature, is then existed 99mg (0.8mmol, 4.0eq) DIEA, 41mg (0.8mmol, 4.0eq) NH is added under stirring condition₄Cl, 73.4mg (0.4mmol, 2.0eq) EDCI and 52mg (0.4mmol, 2.0eq) is stirred to react at 25 DEG C overnight later.Stop reaction, adds Entering 20ml water quenching to go out, repeatedly be extracted with ethyl acetate, saturated common salt water washing organic phase, anhydrous sodium sulfate is dry, it is concentrated under reduced pressure, Crude product EA is recrystallized to give product 50mg.[LCMS(m/z):451(M+H)⁺].¹HNMR(DMSO,400MHz):δppm 10.1 (1H),9.0(1H),7.7(1H),7.6(2H),7.5(1H),7.3(2H),7.2(2),4.7(1H),4.2(2H),3.9(2H), 3.6(2H),3.0(1H),2.8(1H)。

Experimental example 1: biological activity test experiment:

1, compound is to factor X activity suppression test experiments:

Compound 3 times of serial dilutions in dimethyl sulfoxide.Then the compound after diluting uses reaction buffer (50mM again Tris, pH value 8.3；150mM NaCl；2mM CaCl₂；0.005% polysorbas20) 100 times are diluted again.By compound and the tenth because Fluorogenic substrate (the U.S. AnaSpec of 1/2 volume is added after reaction buffer preincubate 15 minutes in sub (final concentration of 0.8nm) Company).Sample is in microplate reader kinetics model 30 degree of read plates Celsius 30 minutes (485nm excitation wavelength and 530nm transmitted waves It is long).The plotted against concentration of signal increased hasten rate and compound, data carry out the Hill equation model of 4 parameters and obtain 50% Inhibition concentration (IC₅₀), concrete outcome is shown in Table 1.

1 factor X activity suppression activity (IC of table₅₀)

Compound number	Factor X inhibitory activity (IC₅₀, μM)	Compound number	Factor X inhibitory activity (IC₅₀, μM)
				Embodiment 1	10	Embodiment 33	0.0008
Embodiment 2	10	Embodiment 34	3.06
				Embodiment 3	2.20	Embodiment 35	0.0028
Embodiment 4	7.60	Embodiment 36	1.23
				Embodiment 5	5.10	Embodiment 37	0.0048
Embodiment 6	10	Embodiment 38	0.016
				Embodiment 7	10	Embodiment 39	0.0006
Embodiment 8	10	Embodiment 40	0.006
				Embodiment 9	10	Embodiment 41	0.006
Embodiment 10	2.50	Embodiment 42	0.0007
				Embodiment 11	0.24	Embodiment 43	0.0019
Embodiment 12	10	Embodiment 44	2.35
				Embodiment 13	10	Embodiment 45	1.9
Embodiment 14	0.32	Embodiment 46	3.66
				Embodiment 15	0.002	Embodiment 47	10
Embodiment 16	0.020	Embodiment 48	10
				Embodiment 17	0.0040	Embodiment 49	2.44
Embodiment 18	0.014	Embodiment 50	2.98
				Embodiment 19	0.022	Embodiment 51	0.0036
Embodiment 20	10	Embodiment 52	0.043
				Embodiment 21	10	Embodiment 53	0.01
Embodiment 22	10	Embodiment 54	0.005
				Embodiment 23	10	Embodiment 55	10
Embodiment 24	10	Embodiment 56	0.009
				Embodiment 25	10	Embodiment 57	0.09
Embodiment 26	0.13	Embodiment 58	0.035
				Embodiment 27	0.0030	Embodiment 59	0.223
Embodiment 28	0.67	Embodiment 60	0.065

Embodiment 29	0.34	Embodiment 61	0.012
				Embodiment 30	0.0002	Embodiment 62	0.009
Embodiment 31	0.70	Embodiment 63	1.3
				Embodiment 32	5.81

2, pharmacokinetics is tested:

1.33mg compound is accurately weighed, the DMSO of 0.333mL is added, is vortexed 1 minute, then ultrasound 2 minutes is added 2.66mL PEG400 is vortexed 1 minute, and ultrasound 2 minutes is eventually adding 3.658mL physiological saline, is vortexed 1 minute, 2 points of ultrasound Clock obtains clear solution, and compound concentration 0.2mg/mL is colorless cleared solution (pH~8) to be used for the 1st group of intravenously administrable.

5.46mg compound is accurately weighed, the DMSO of 0.546mL is added, is vortexed 1 minute, then ultrasound 2 minutes is added 4.368mL PEG400 is vortexed 1 minute, and ultrasound 2 minutes is eventually adding 6.006mL physiological saline, is vortexed 1 minute, 2 points of ultrasound Clock obtains clear solution, and compound concentration 0.5mg/mL is colorless cleared solution (pH~9) to be used for the 2nd group of oral administration.

Every part of test solution keep sample 200 μ L with HPLC detect administration solution concentration.

Rat 6, it is divided to two groups of intravenous injection administration and oral administrations respectively.Fasting 12-16 hours before being administered orally.Administration Recovery in 4 hours is fed afterwards.Detailed clinical observation is carried out before administration and after administration, it is no abnormal.

It takes a blood sample respectively to every animal, animal blood taking time point are as follows: vein and oral: before administration, 5min after administration, 15min, 30min, 1h, 2h, 3h, 5h, 8h and for 24 hours；About 0.25mL blood is adopted through jugular vein, heparin sodium is anticoagulant.Blood sample Acquisition is placed on ice, and centrifugal separation plasma (centrifugal condition: 8000 revs/min, 6 minutes, 2-8 DEG C) within 30 minutes. - 80 DEG C are deposited in front of the plasma analysis of collection.

Sample (- 80 DEG C) are taken out from refrigerator, room temperature is vortexed 30 seconds after dissolving naturally, takes 50 μ L samples to 1.5mL centrifugation Guan Zhong.It is added 250 μ L inner mark solutions (200ng/mL orinase methanol solution), is centrifuged 5 minutes (15000 after being vortexed 60 seconds Rev/min), 200 μ L supernatants to 96 orifice plates are taken, sample introduction is in ultra performance liquid chromatography system (Waters company, ACQUITY UPLC)-mass spectrograph (API 4000, Applied biosystems, electric spray ion source (ESI), series connection quadrupole rod quality point Parser) it is analyzed.

Data handle through Analyst software data processing system and obtain pharmacokinetics results.

It the results are shown in Table 2

2 part of compounds pharmacokinetic data of table

Compound number	T_max	t_1/2	Cl_zL/hr/kg	V_zL/kg	F
						FXa-14	2.00±0.00	1.46~1.70	0.88±0.17	1.37±0.10	2.19 ± 0.24%
FXa-84	2.33±0.58	2.96~3.78	0.38±0.08	0.71±0.11	14.75 ± 4.52%
						FXa-49	2.86±4.46	1.58~2.99	1.27±0.59	2.83±1.08	0.78 ± 0.16%
FXa-86	1.50±0.87	0.77~0.92	1.29±0.24	1.54±0.31	0.31 ± 0.11%
						FXa-93	0.25±0.00	0.87~1.39	2.40±0.27	1.30±0.13	6.78 ± 1.63%
FXa-61	2.00±0.0	2.12~2.85	1.69±0.63	2.71±1.05	1.82 ± 0.37%.
						FXa-20	0.25±0.00	1.53~3.45	3.61±0.48	2.19±0.08	5.39 ± 1.06%

Table 2 as the result is shown compound in the intracorporal absorption of rat animal, drug distribution, accretion rate, excretion and life Situations such as object availability.

Although above having used general explanation, specific embodiment and test, the present invention is made to retouch in detail It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art 's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed Range.

Claims

1. one kind has following any structure compound represented or its pharmaceutically acceptable salt:

2. the preparation containing compound described in claim 1, which is characterized in that the preparation is by compound and pharmaceutically acceptable Carrier composition.

3. preparation according to claim 2, which is characterized in that the preparation is tablet, capsule, granule, injection.

4. preparation according to claim 2, which is characterized in that the pharmaceutically acceptable carrier is selected from filler, collapses Solve agent, adhesive, lubricant.

5. compound described in claim 1 or the described in any item preparations of claim 2-4 treat and prevent thrombus in preparation Application in the drug of formation.

6. application according to claim 5, which is characterized in that the thrombus be non-valvular atrial fibrillation patient occur stroke or Systemic embolism, hip joint and knee prosthesis postoperative patient person deep vein thrombosis and pulmonary embolism.