Ester derivatives of multi-substituted 4-methylamino-benzamidine as well as preparation method and application of ester derivatives

[0096] Example 1

[0097] 3-{1-Methyl-2-[N-(4-(N-pentyloxycarbonyl)amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid}-( Synthesis and preparation of N-2-pyridyl)amido-propionic acid-2-(n-heptadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester (compound 1)

[0098]

[0099] (1) Synthesis of 3-(N-2-pyridylamino)-propionic acid ethyl ester

[0100]

[0101] Under the protection of nitrogen, add ethyl acrylate (27.5g, 0.275mol) to 2-aminopyridine (22.5g, 0.25mol), stir and reflux at a temperature higher than 100°C for 24h, filter out the precipitate, concentrate the residue and use a silica gel column Column purification gave 3-(N-2-pyridylamino)-propionic acid ethyl ester (34.9 g, yield 72%) as a white solid. Mass Spectrum (ESI-MS): 195.1 (M+H) + , 217.3 (M+Na) +;C 10 h 14 N 2 o 2 (194).

[0102] (2) Synthesis of 4-cyanoanilino-acetic acid

[0103]

[0104] Add 150ml of water to 4-cyanoaniline (6.0g, 0.05mol) and 1-chloroacetic acid (10g, 0.11mol), heat to reflux until a large amount of yellow solid precipitates, filter at room temperature, and wash with water, absolute ethanol, Rinse with anhydrous ether to obtain 4-cyanoanilino-acetic acid (6.4 g, yield 73%) as a yellow solid. Mass Spectrum (ESI-MS): 177.3 (M+H) + , 199.2 (M+Na) +;C 9 h 8 N 2 o 2 (176).

[0105] (3) Synthesis of 4-methylamino-3-nitro-benzoic acid

[0106]

[0107] Add 150mL of 25%-30% methylamine aqueous solution to 3-nitro-4-chlorobenzoic acid (25g, 0.124mol), and make the system react at a temperature higher than 100°C for 5-6h. After concentration under reduced pressure, glacial acetic acid was added to adjust the pH to 4-5. After standing overnight, a large amount of yellow solid precipitated, the solution was filtered off, and the precipitate was recrystallized at least twice with 95% ethanol to obtain a yellow solid 4-aminomethyl-3-nitro-benzoic acid (18.5g, yield 76% ). Mass Spectrum (ESI-MS): 197.2 (M+H) + , 219.4 (M+Na) +;C 8 h 8 N 2 o 4 (196).

[0108] (4) Synthesis of 4-methylamino-3-nitro-benzoyl chloride

[0109]

[0110] Dissolve 4-methylamino-3-nitro-benzoic acid (11.6g, 0.059mol) in 150mol thionyl chloride, reflux for 1.5h, concentrate under reduced pressure and add 50-60ml of CH 2 Cl 2 , so that it is completely dissolved to obtain the CH of 4-methylamino-3-nitro-benzoyl chloride 2 Cl 2 solution. Mass Spectrum (ESI-MS): 215.3 (M+H) +;C 8 h 7 ClN 2 o 3 (214).

[0111] (5) Synthesis of ethyl 3-(2-nitro-1-methylamino-phenyl-4-yl)-carboxylic acid-(N-2-pyridyl)-amidopropionate

[0112]

[0113] Dissolve ethyl 3-(N-2-pyridylamino)-propionate (9.7 g, 0.05 mol) in 30 ml CH 2 Cl 2 and 30ml triethylamine, slowly add 4-methylamino-3-nitro-benzoyl chloride CH at room temperature 2 Cl 2 solution. The mixed system was reacted at room temperature for 12 hours, the precipitate was filtered off, and the filtrate was concentrated and purified with a silica gel column to obtain a yellow oily liquid 3-(2-nitro-1-methylamino-benzene-4-yl)-carboxylic acid-( N-2-Pyridyl)amido-propionic acid ethyl ester (15.4 g, 83% yield). Mass Spectrum (ESI-MS): 373.1 (M+H) + , 395.2 (M+Na) +;C 18 h 20 N 4 o 5 (372).

[0114] (6) Synthesis of 3-(1-methylamino-2-amino-phenyl-4-yl)-carboxylic acid-(N-2-pyridyl)amido-propionic acid ethyl ester

[0115]

[0116] Dissolve 3-(2-nitro-1-methylamino-phen-4-yl)-carboxylic acid-(N-2-pyridyl)amido-propionic acid ethyl ester (10.0g, 0.027mol) in 120ml without In water ethanol, add 1.0g 10%Pd/C, react continuously at 30atm. for 20h, filter and concentrate to get 3-(1-methylamino-2-amino-benzene-4-yl)-carboxylic acid-(N -2-pyridyl)amido-propionic acid ethyl ester (7.8g, yield 85%). Mass Spectrum (ESI-MS): 343.4 (M+H) + , 365.3 (M+Na) +;C 18 h 22 N 4 o 3 (342).

[0117] (7) 3-(1-methyl-2-(4-cyano-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-2-pyridyl)amido)propionic acid Synthesis of Ethyl Ester

[0118]

[0119] Dissolve 4-cyanoanilino-acetic acid (2.32g, 13.2mmol) in 80ml DMF, add 1-hydroxybenzotriazole (HOBT) (1.96g, 14.5mmol), add 1-ethyl -3-(3-Dimethylaminopropyl)carbodiimide hydrochloride (EDCI) (2.77g, 14.5mmol) stirred for 45min, slowly rose to room temperature, added 3-(1-methylamino-2-amino- Phenyl-4-yl)-carboxylic acid-(N-2-pyridyl)amidopropanoic acid ethyl ester (5.0 g, 14.5 mmol). React overnight at room temperature, concentrate, dilute with a large amount of ethyl acetate, wash with saturated brine three times, wash with Na 2 SO 4 After drying and concentration, the crude product was refluxed in 60ml of acetic acid for 1.5h. After concentration, it was alkalized by adding 1.5N ammonia water, extracted three times with ethyl acetate (60mL×3), and the organic phase was washed once with saturated brine, washed with Na 2 SO 4 After drying and concentration, the crude product was purified layer by layer with silica gel to obtain an amorphous solid 3-(1-methyl-2-(4-cyano-phenylaminomethyl)-benzimidazol-5-yl-carboxylate Acid-(N-2-pyridyl)amido)propanoic acid ethyl ester (6.4 g, yield 83%). Mass Spectrum (ESI-MS): 482.1 (M+H) + , 505.4 (M+Na) +;C 27 h 26 N 6 o 3 (482).

[0120] (8) 3-(1-Methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-2-pyridyl)amido)propanoic acid Synthesis of Ethyl Ester

[0121]

[0122] 3-(1-Methyl-2-(4-cyano-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-2-pyridyl)amido)propanoic acid ethyl ester (11.1g, 23mmol) was dissolved in 200ml of absolute ethanol, passed through dry HCl gas until saturated, and stirred at room temperature for 12h. After concentration under reduced pressure, 200 ml of saturated NH3 absolute ethanol was added, reacted at room temperature for 5 h, concentrated, and the crude product was purified by silica gel column chromatography to obtain an amorphous solid 3-(1-methyl-2-(4-amidino-benzene (aminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-2-pyridyl)amido)propionic acid ethyl ester (9.3 g, yield 81%). Mass Spectrum (ESI-MS): 500.2 (M+H) + , 522.3 (M+Na) +;C 27 h 29 N 7 o 3 (499).

[0123] (9) 3-(1-Methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-2-pyridyl)amido)propionic acid Synthesis

[0124]

[0125] Ethyl 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-2-pyridyl)amido)propionate (8.0g, 16mmol) dissolved in 160ml absolute ethanol 80ml H 2O, added sodium hydroxide (1.92g, 48mmol), stirred at room temperature for 3h. Dilute it with 400ml of water, add an appropriate amount of acetic acid to neutralize it, let it stand still, a large amount of white precipitate precipitates out, filter off the solution, rinse with water, absolute ethanol, and anhydrous ether in sequence, and obtain a white solid 3-(1-formazol after drying) yl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-2-pyridyl)amido)propionic acid (6.3g, yield 84%) . Mass Spectrum (ESI-MS): 472.1 (M+H) + , 494.2 (M+Na) +;C 25 h 25 N 7 o 3 (471).

[0126] (10) 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid)-(N-2-pyridyl)amido-propane Synthesis of Acid-2-(n-heptadecylcarbonyloxy)glycerophosphorylcholine-3-yl-Ester

[0127]

[0128] Add 30-40ml of dichloromethane, 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid- (N-2-pyridyl)amido)propionic acid 5.0g, 2-(n-heptadecylcarbonyloxy)-glycerophosphorylcholine 5.6g, dicyclohexylcarbodiimide (DCC) 2.1g, 4-N,N-Dimethylaminopyridine (DMAP) 70mg, stirred at room temperature for 3h (TLC detection end point), suction filtered after the reaction was complete, the filtrate was washed with 3×20mL20% sodium carbonate solution, and then saturated sodium chloride The solution was washed until neutral, dried over anhydrous magnesium sulfate, and the solvent was evaporated to obtain a yellow solid, which was recrystallized with 95% ethanol to obtain a white solid 3-(1-methyl-2-(4-amidino-phenyl Aminomethyl)-benzimidazol-5-yl-carboxylic acid)-(N-2-pyridyl)amido-propionic acid-2-(n-heptadecylcarbonyloxy)glycerophosphorylcholine-3 -yl-ester 7.6 g, yield 73.4%. Mass Spectrum (ESI-MS): 977.4 (M+H) + , 999.5 (M+Na) +;C 51 h 77 N 8 o 9 P(976).

[0129] (11) 3-{1-methyl-2-[N-(4-(N-pentyloxycarbonyl)amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid Synthesis of}-(N-2-pyridyl)amido-propionic acid-2-(n-heptadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester (Compound 1)

[0130]

[0131] 3-(1-Methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid)-(N-2-pyridyl)amido-propionic acid- 2-(n-Heptadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester (9.8 g, 10 mmol) was dissolved in 250 ml THF and 50 ml H 2 O, potassium hydroxide (2.1 g, 15 mmol) was added, and stirred at room temperature for 15 min. n-Pentyl chloroformate (1.81 g, 11 mmol) was added slowly and stirring was continued for 2 h. Concentrated, extracted three times with ethyl acetate (20ml×3), washed with Na 2 SO 4 After drying and concentration, the crude product was purified by silica gel column chromatography to obtain the final product 3-{1-methyl-2-[N-(4-(N-pentyloxycarbonyl)amidinophenyl) as a white solid -Aminomethyl]-benzimidazol-5-yl-carboxylic acid}-(N-2-pyridyl)amido-propionic acid-2-(n-heptadecylcarbonyloxy)glycerophosphorylcholine- 3-yl-ester (compound 1, 8.3 g, 76% yield). Mass Spectrum (ESI-MS): 1091.4 (M+H) + , 1113.6 (M+Na) +;C 51 h 77 N 8 o 9 P(1090). High resolution mass spectrometry (HR-ESI-MS): 1113.6249 (M+Na) + , calculated value: 1113.6233).

[0132] 1 H NMR (600MHz, DMSO-d 6 )δ: 0.88(m,6H),1.27~1.33(m,32H),1.58(m,2H),1.67(m,2H),2.25(t,J=7.6Hz,2H),2.68(t,J =14.5Hz,2H),3.37(s,9H),3.61(t,J=14.2Hz,2H),3.77(s,3H),3.95(m,2H),4.22(t,J=14.5Hz,2H ),4.59(d,J=5.5Hz,2H),4.30(m,2H),4.50(m,4H),5.20(m,1H),6.76(d,J=9.0Hz,2H),6.88(d ,J=7.5Hz,1H),6.97(s,1H),7.10~7.13(m,1H),7.15(dd,J1=8.5Hz,J2=1.5Hz,1H),7.40(d,J=8.5Hz ,1H),7.47(d,J=1.5Hz,1H),7.54(td,J1=10.5Hz,J2=2.0Hz,1H),7.80(d,J=8.5Hz,2H),8.39(dq,J1 =5.0Hz, J2=1.7Hz, 1H), 8.90(br s, 2H).

[0133] Example 2

[0134] 3-{1-Methyl-2-[N-(4-(N-Hexyloxycarbonyl)amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid}-(N Synthesis and preparation of -phenyl)amido-propionic acid-3-(n-octadecylcarbonyloxy)glycerophosphorylcholine-2-yl-ester (compound 2)

[0135]

[0136] (1) Synthesis of 3-(N-phenylamino)-ethyl propionate

[0137]

[0138] Under the protection of nitrogen, add ethyl acrylate (27.5g, 0.275mol) to aniline (23.3g, 0.25mol), add 10ml of absolute ethanol and 10ml of triethylamine, stir and reflux at a temperature higher than 100°C for 24h, filter out the precipitate The residue was concentrated and purified with a silica gel column to obtain light red solid 3-(N-phenylamino)-propionic acid ethyl ester (36.7g, yield 76%). Mass Spectrum (ESI-MS): 194.1 (M+H) + , 216.3 (M+Na) +;C 11 h 15 NO 2 (193).

[0139] (2) Synthesis of ethyl 3-(2-nitro-1-methylamino-phenyl-4-yl)-carboxylic acid-(N-phenyl)amidopropionate

[0140]

[0141] 3-(N-Phenylamino)-propionic acid ethyl ester (9.7g, 0.05mol) was dissolved in 30ml CH 2 Cl 2 and 30ml triethylamine, slowly add 4-methylamino-3-nitro-benzoyl chloride CH at room temperature 2 Cl 2 solution. The mixed system was reacted at room temperature for 12 hours, the precipitate was filtered off, and the filtrate was concentrated and purified with a silica gel column to obtain a yellow oily liquid 3-(2-nitro-1-methylamino-benzene-4-yl)-carboxylic acid-( N-Phenyl)amido-propionic acid ethyl ester (15.4 g, 83% yield). Mass Spectrum (ESI-MS): 372.2 (M+H) + , 394.3 (M+Na) +;C 19 h 21 N 3 o 5 (371).

[0142] (3) Synthesis of 3-(1-methylamino-2-amino-phenyl-4-yl)-carboxylic acid-(N-2-pyridyl)amido-propionic acid ethyl ester

[0143]

[0144] Dissolve ethyl 3-(2-nitro-1-methylamino-phen-4-yl)-carboxylic acid-(N-phenyl)-amidopropionate (10.0 g, 0.027 mol) in 120 ml absolute ethanol Add 1.0g of 10%Pd/C, continue the reaction at 30atm. for 20h, filter and concentrate to get 3-(1-methylamino-2-amino-benzene-4-yl)-carboxylic acid-(N-benzene base) amido-ethyl propionate (8.0 g, yield 87%). Mass Spectrum (ESI-MS): 342.2 (M+H) + , 364.2 (M+Na) +;C 19 h 23 N 3 o 3 (341).

[0145] (4) Ethyl 3-(1-methyl-2-(4-cyano-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-phenyl)amido)propionate Synthesis

[0146]

[0147] Dissolve 4-cyanoanilino-acetic acid (2.32g, 13.2mmol) in 80ml DMF, add 1-hydroxybenzotriazole (HOBT) (1.96g, 14.5mmol), and add 1-ethanediol at -20°C Dimethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) (2.77g, 14.5mmol) was stirred for 45min, slowly raised to room temperature, and 3-(1-methylamino-2-amino -Phenyl-4-yl)-carboxylic acid-(N-phenyl)-amidopropionic acid ethyl ester (5.0 g, 14.5 mmol). React overnight at room temperature, concentrate, dilute with a large amount of ethyl acetate, wash with saturated brine three times, wash with Na 2 SO 4 After drying and concentration, the crude product was refluxed in 60ml of acetic acid for 1.5h. After concentration, it was alkalized by adding 1.5N ammonia water, extracted three times with ethyl acetate (60mL×3), and the organic phase was washed once with saturated brine, washed with Na 2 SO 4 After drying and concentration, the crude product was purified layer by layer with silica gel to obtain an amorphous solid 3-(1-methyl-2-(4-cyano-phenylaminomethyl)-benzimidazol-5-yl-carboxylate Acid-(N-phenyl)-amido)propionate ethyl ester (5.1 g, 81% yield). Mass Spectrum (ESI-MS): 482.2 (M+H) + , 504.2 (M+Na) +;C 28 h 27 N 5 o 3 (481).

[0148] (5) Ethyl 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-phenyl)amido)propionate Synthesis

[0149]

[0150] 3-(1-Methyl-2-(4-cyano-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-phenyl)amido)propionic acid (11.0 g, 23mmol) was dissolved in 200ml of absolute ethanol, passed through dry HCl gas until saturated, and stirred at room temperature for 12h. After concentrating under reduced pressure, add 200ml saturated NH 3 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzo Imidazol-5-yl-carboxylic acid-(N-phenyl)amido)propionic acid (8.9 g, 78% yield). Mass Spectrum (ESI-MS): 499.1 (M+H) + , 521.2 (M+Na) +;C 28 h 30 N 6 o 3 (498).

[0151] (6) Synthesis of 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-phenyl)amido)propionic acid

[0152]

[0153] Ethyl 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-phenyl)amido)propionate (8.0 g, 16mmol) dissolved in 160ml absolute ethanol 80ml H 2 O, added sodium hydroxide (1.92g, 48mmol), stirred at room temperature for 3h. Dilute it with 400ml of water, add an appropriate amount of acetic acid to neutralize it, let it stand still, a large amount of white precipitate precipitates out, filter off the solution, rinse with water, absolute ethanol, and anhydrous ether in sequence, and obtain a white solid 3-(1-formazol after drying) yl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid-(N-phenyl)amido)propionic acid (6.0 g, yield 80%). Mass Spectrum (ESI-MS): 471.2 (M+H) + , 493.3 (M+Na) +;C 26 h 26 N 6 o 3 (470).

[0154] (7) 3-(1-Methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid)-(N-phenyl)amido-propionic acid- Synthesis of 3-(n-octadecylcarbonyloxy)glycerophosphorylcholine-2-yl-ester

[0155]

[0156] Add 30-40ml of dichloromethane, 3-(1-methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid- (N-phenyl)amido)propionic acid 5.0g, 2-(n-octadecylcarbonyloxy)-glycerophosphorylcholine 5.6g, dicyclohexylcarbodiimide (DCC) 2.1g, 4- N,N-Dimethylaminopyridine (DMAP) 70 mg, stirred at room temperature for 3 h (TLC detection end point), suction filtered after the reaction was complete, the filtrate was washed with 3×20 mL of 20% sodium carbonate solution, and then washed with saturated sodium chloride solution To neutrality, after drying over anhydrous magnesium sulfate, the solvent was evaporated to obtain a yellow solid, which was recrystallized with 95% ethanol to obtain a white solid 3-(1-methyl-2-(4-amidino-phenylaminomethanol) base)-benzimidazol-5-yl-carboxylic acid)-(N-phenyl)amido-propionic acid-3-(n-octadecylcarbonyloxy)glycerophosphorylcholine-2-yl-ester 7.6g, yield 72.9%. Mass Spectrum (ESI-MS): 990.6 (M+H) + , 1012.7 (M+Na) +;C 53 h 80 N 7 o 9 P(989).

[0157] (8) 3-{1-Methyl-2-[N-(4-(N-Hexyloxycarbonyl)amidinophenyl)-aminomethyl]-benzimidazol-5-yl-carboxylic acid} Synthesis of -(N-phenyl)amido-propionic acid-3-(n-octadecylcarbonyloxy)glycerophosphorylcholine-2-yl-ester

[0158]

[0159] 3-(1-Methyl-2-(4-amidino-phenylaminomethyl)-benzimidazol-5-yl-carboxylic acid)-(N-phenyl)amido-propionic acid-3- (n-octadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester (9.8 g, 10 mmol) was dissolved in 250 ml THF and 50 ml H 2 O, potassium hydroxide (2.1 g, 15 mmol) was added, and stirred at room temperature for 15 min. Add n-hexyl chloroformate (1.81 g, 11 mmol) slowly and continue stirring for 2 h. Concentrated, extracted three times with ethyl acetate (20ml×3), washed with Na 2 SO 4 After drying and concentration, the crude product was purified by silica gel column chromatography to obtain the final product 3-{1-methyl-2-[N-(4-(N-hexaneoxycarbonyl)amidinophenyl)- Aminomethyl]-benzimidazol-5-yl-carboxylic acid}-(N-phenyl)amido-propionic acid-3-(n-octadecylcarbonyloxy)glycerophosphorylcholine-2-yl -Ester (compound 2, 8.2g, yield 73%). Mass Spectrum (ESI-MS): 1118.7 (M+H) + , 1140.6 (M+Na) +;C 60 h 92 N 7 o 11 P(1117). High resolution mass spectrometry (HR-ESI-MS): 1140.6530 (M+Na) + , calculated value: 1140.6572).

[0160] 1 H NMR (600MHz, DMSO-d 6 )δ: 0.90(m,6H),1.25~1.34(m,36H),1.58(m,2H),1.67(m,2H),2.26(t,J=7.6Hz,2H),2.70(t,J =14.5Hz,2H),3.38(s,9H),3.62(t,J=14.2Hz,2H),3.78(s,3H),3.97(m,2H),4.23(t,J=14.5Hz,2H ),4.61(d,J=5.5Hz,2H),4.32(m,2H),4.52(m,4H),5.22(m,1H),6.76(d,J=9.2Hz,2H),6.82(d ,J=7.6Hz,1H),6.92(s,1H),7.12(t,J=7.2Hz,1H),7.24(t,J=7.2Hz,2H),7.40(d,J=8.4Hz,1H ), 7.47 (d, J=1.5Hz, 1H), 7.64 (d, J=7.2Hz, 2H), 7.80 (d, J=8.4Hz, 2H), 8.93 (br s, 2H).

[0161] Example 3

[0162] 3-{2-[N-(4-(N-Butoxycarbonyl)amidinophenyl)-aminomethyl]-benzothiazol-5-yl-carboxylic acid-(N-phenyl)}-amide Synthesis and preparation of -propionic acid-2-(n-hexadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester (compound 3)

[0163]

[0164] (1) Synthesis of 4-fluoro-3-methoxyacetamido-ethyl benzoate

[0165]

[0166] 17.4g (94.9mmol) 3-amino-4-fluoro-benzoic acid ethyl ester (refer to L.S.Fosdick, A.F.Dodds, J.Amer Chem.Soc.65,2305 (1943)) and 9.67ml (11.47g, 105.4mmol ) A solution of methoxyacetyl chloride in 310 ml of chlorobenzene was stirred at 50°C for 2 hours and then refluxed for 30 minutes. Concentrate under reduced pressure and evaporate to dryness, and purify by silica gel column chromatography (dichloromethane/ethanol=100:1) to obtain the oily product 3-amino-4-fluoro-ethyl benzoate, which solidifies into a solid after a few days (20.1g, yield rate of 83%). R f Value: 0.38 (silica gel: dichloromethane/ethanol = 19:1). Mass Spectrum (ESI-MS): 256.1 (M+H) + , 278.1 (M+Na) +;C 12 h 14 FNO 4 (255).

[0167] (2) Synthesis of ethyl 2-methoxymethyl-benzothiazole-5-carboxylate

[0168]

[0169] 18.6g (72.5mmol) of 4-fluoro-3-methoxyacetamido-ethyl benzoate and 13.0g (32.2mmol) of Lawesson's reagent (Lawesson's reagent) were refluxed in 90ml of toluene for 6 hours, and then mixed with 6.2g of Lawesson's reagent Sen's reagent was mixed and heated to 120°C for 6 hours. After the solvent was changed to xylene, the mixture was heated at 180° C. for 8 hours in a pressurized reactor. Then concentrated under reduced pressure to remove the solvent, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether=5:95) to obtain yellow crystal 2-methoxymethyl-benzothiazole-5-carboxylic acid ethyl ester (13.6g, yield 75%). R f Value: 0.55 (silica gel: ethyl acetate/petroleum ether = 3:7). Mass Spectrum (ESI-MS): 252.1 (M+H) + , 274.1 (M+Na) +;C 12 h 13 NO 3 S (251).

[0170] (3) Synthesis of 2-methoxymethyl-benzothiazole-5-carboxylic acid

[0171]

[0172] A mixture of 13.0 g (51.8 mmol) of ethyl 2-methoxymethyl-benzothiazole-5-carboxylate and 100 ml of 2N sodium hydroxide solution was stirred at room temperature into 370 ml of ethanol and stirring was continued for 1 hour. Concentrate under reduced pressure to obtain a crude product, which is dissolved in 125 ml of water, washed with 310 ml of diethyl ether, and the aqueous phase is acidified with concentrated hydrochloric acid while cooling with ice. Suction filtration, washing with water, and drying gave the beige solid product 2-methoxymethyl-benzothiazole-5-carboxylic acid (9.1 g, yield 79%). R f Value: 0.12 (silica gel: dichloromethane/ethanol = 29:1). Mass Spectrum (ESI-MS): 224.0 (M+H) + , 246.0 (M+Na) +;C 10 h 9 NO 3 S (223).

[0173] (4) Synthesis of 2-methoxymethyl-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl)-amide

[0174]

[0175]A suspension of 10.0 g (44.6 mmol) of 2-methoxymethyl-benzothiazole-5-carboxylic acid in 370 ml of dichloromethane was mixed with 10 ml (136.4 mmol) of sulfuryl chloride and refluxed for 1 hour. After 20 minutes the solid dissolved. The solvent was distilled off under reduced pressure to obtain a crude product, dissolved in dichloromethane and evaporated to dryness, dissolved again, evaporated to dryness once, dissolved in 310ml tetrahydrofuran, added dropwise to 8.7g (44.6mmol) N-(2-ethoxycarbonyl Ethyl) aniline, 18.6ml (130.2mmol) triethylamine and 310ml tetrahydrofuran mixed solution, stirred overnight at room temperature. After evaporated to dryness under reduced pressure, it was dissolved in 30 ml of dichloromethane, the solution was washed with water, and dried over sodium sulfate. After concentration under reduced pressure, it was purified by silica gel column chromatography (gradient elution, dichloromethane/ethanol=98.5:1.5 to 80:20) to obtain the brown oily product 2-methoxymethyl-benzothiazole-5-carboxylic acid -N-Phenyl-N-(2-ethoxycarbonylethyl)-amide (12.8 g, 72% yield). R f Value: 0.40 (silica gel: ethyl acetate/petroleum ether = 1:1). Mass Spectrum (ESI-MS): 399.1 (M+H) + , 437.1 (M+K) +;C 21 h 22 N 2 o 4 S (398).

[0176] (5) 2-[N-(4-cyanophenyl)-aminomethyl]-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl)-amide Synthesis

[0177]

[0178] 12.7g (31.9mmol) of 2-methoxymethyl-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl)amide was dissolved in 35.3ml (35.3mmol) 1 M dichloromethane solution of boron tribromide was redissolved in 370ml of dichloromethane, and stirred at room temperature for 16 hours. Then wash with 40ml saturated sodium bicarbonate solution, dry the organic phase with sodium sulfate, and evaporate to dryness to obtain the intermediate 2-bromomethyl-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxy Carbonylethyl)-amide (14.9g). This intermediate was dissolved in 5.0 ml of N,N-diisopropyl-ethylamine and mixed with 4.0 g (33.5 mmol) of 4-aminobenzonitrile. After heating at 130°C for 1 hour, evaporate to dryness under reduced pressure, purify by silica gel column chromatography (gradient, ethyl acetate/petroleum ether = 1:3 to 1:1), and concentrate the eluent under reduced pressure to obtain an orange foamy product 2-[N-(4-Nitrylphenyl)-aminomethyl]-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl)-amide (6.8g , yield 44%). R f Value: 0.35 (silica gel: ethyl acetate/petroleum ether = 7:3). Mass Spectrum (ESI-MS): 485.2 (M+H) + , 486.2 (M+2H) ++;C 27 h 24 N 4 o 3 S (484).

[0179] (6) 2-[N-(4-amidinophenyl)-aminomethyl]-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl)-amide Synthesis

[0180]

[0181] 6.8g (14mmol) 2-[N-(4-cyanophenyl)-aminomethyl]-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl ) amide in 620 ml of ethanol saturated with hydrogen chloride was stirred at 0°C for 5 hours and then at room temperature until no starting reactant was detected by thin layer chromatography. Then the solvent was distilled off at a water bath temperature not exceeding 30°C, and the oily product was dissolved in 620 ml of absolute ethanol and mixed with 10 g (136.4 mmol) of ammonium carbonate. After stirring at room temperature for 18 hours, it was evaporated to dryness under reduced pressure, and purified by column chromatography on a silica gel column (gradient: water/methanol=19:1 to 4:1), and the eluent was evaporated to dryness under reduced pressure to obtain a white foamy product 2-[N-(4-Amidinophenyl)-aminomethyl]-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl)-amide (4.4g , yield 63%). R f Value: 0.19 (silica gel: dichloromethane/ethanol = 3:7); mass spectrometry (ESI-MS): (M+H) + =502.2, 524.2 (M+Na) +;C 27 h 27 N 5 o 3 S(501).

[0182] (7) 2-[N-(4-amidinophenyl)-aminomethyl]-benzothiazol-5-yl-carboxylic acid-N-phenyl-N-(2-carboxyethyl)-amide synthesis

[0183]

[0184] 28g (52mmol) 2-[N-(4-amidinophenyl)-aminomethyl]-benzothiazole-5-carboxylic acid-N-phenyl-N-(2-ethoxycarbonylethyl) The -amide was dissolved in 930 ml of ethanol, mixed with 20 ml of 2N sodium hydroxide solution, and stirred at room temperature for 4 hours. The mixture was then acidified with 30 ml of 2N hydrochloric acid, and the solvent was distilled off. The resulting crude product was dissolved in 50 ml of dichloromethane/ethanol (2:1), and filtered to remove insoluble sodium chloride. After the solvent was removed by distillation under reduced pressure, the yellow foamy solid product 2-[N-(4-amidinophenyl)-aminomethyl]-benzothiazol-5-yl-carboxylic acid-N-phenyl-N-( 2-carboxyethyl)-amide (16 g, 67% yield). R f Value: 0.47 (silica gel: methanol/5% aqueous sodium chloride = 6:4). Mass Spectrum (ESI-MS): (M+H) + =474.2, 496.2 (M+Na) +;C 25 h 23 N 5 o 3 S (473).

[0185] (8) 3-(2-(N-(4-amidinophenyl)-aminomethyl)-benzothiazol-5-yl-carboxylic acid)-(N-phenyl)amido-propionic acid-2 -Synthesis of (n-hexadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester

[0186]

[0187] Add 30-40ml of dichloromethane, 2-[N-(4-amidinophenyl)-aminomethyl]-benzothiazole-5-carboxylic acid-N-phenyl-N -(2-carboxyethyl)-amide 5.0g, 2-(n-hexadecylcarbonyloxy)-glycerophosphorylcholine 5.6g, dicyclohexylcarbodiimide (DCC) 2.1g, 4-N , N-dimethylaminopyridine (DMAP) 70mg, stirred at room temperature for 3h (TLC detection end point), suction filtered after the reaction was complete, the filtrate was washed with 3×20mL 20% sodium carbonate solution, and then washed with saturated sodium chloride solution To neutrality, after drying over anhydrous magnesium sulfate, the solvent was evaporated to obtain a yellow solid, which was recrystallized with 95% ethanol to obtain a white solid 3-(2-(N-(4-amidinophenyl)-aminomethyl )-benzothiazol-5-yl-carboxylic acid)-(N-phenyl)amido-propionic acid-2-(n-hexadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester 7.2 g, yield 71%. Mass Spectrum (ESI-MS): (M+H) + =965.5; C 50 h 73 N 6 o 9 PS (964).

[0188] (9) 3-{2-[N-(4-(N-butoxycarbonyl)amidinophenyl)-aminomethyl]-benzothiazol-5-yl-carboxylic acid}-(N-phenyl ) Synthesis of amido-propionic acid-2-(n-hexadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester (compound 3)

[0189]

[0190] 6.2 g (6.4 mmol) of 3-{2-[N-(4-amidinophenyl)-aminomethyl]-benzothiazol-5-yl-carboxylic acid}-(N-phenyl)amido- Propionate-2-(n-hexadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester was dissolved in a mixed solution of 120ml tetrahydrofuran and 30ml water, and 1.77g (12.8mmol) potassium carbonate and 931mg ( 6.8mmol) of n-butyl chloroformate, stirred at room temperature for 2 hours, evaporated to dryness under reduced pressure and mixed with 150ml of saturated saline, the resulting solution was extracted 3 times with 60ml of dichloromethane each time, and the extract was dried over sodium sulfate. After being evaporated to dryness under reduced pressure, it was purified on a silica gel column to obtain the final product 3-{2-[N-(4-(N-butoxycarbonyl)amidinophenyl)-aminomethyl]-benzothiazol-5-yl -Carboxylic acid}-(N-phenyl)amido-propionic acid-2-(n-hexadecylcarbonyloxy)glycerophosphorylcholine-3-yl-ester (compound 3, 5.3g, yield 78 %). Mass Spectrum (ESI-MS): (M+H) + =1065.5, (M+H+Na) ++ =1088.5, (M+2H) + =1066.5,C 55 h 81 N 6 o 11 PS(1064). High resolution mass spectrometry (HR-ESI-MS): 1087.5359 (M+Na) + , calculated value: 1087.5375).

[0191] 1 H NMR (600MHz, DMSO-d 6 )δ: 0.88(m,6H),1.22~1.31(m,28H),1.56(m,2H),1.68(m,2H),2.24(t,J=7.5Hz,2H),2.68(t,J =14.6Hz,2H),3.37(s,9H),3.60(t,J=14.2Hz,2H),3.96(m,2H),4.21(t,J=14.6Hz,2H),4.58(d,J =5.5Hz,2H),4.28(m,2H),4.48(m,4H),5.19(m,1H),6.75(d,J=9.0Hz,2H),6.83(d,J=7.4Hz,1H ),6.94(s,1H),7.10(t,J=7.4Hz,1H),7.22(t,J=7.4Hz,2H),7.36(d,J=8.2Hz,1H),7.46(d,J =1.5Hz, 1H), 7.65(d, J=7.4Hz, 2H), 7.78(d, J=8.2Hz, 2H), 8.90(br s, 2H).