CN101318933B - Isoquinolinium compounds with antithrombotic activity, preparation method and application thereof - Google Patents

Abstract

The invention discloses compounds with anti-thrombus activity shown in a formula I, a preparation method thereof and an application of the compounds as antithrombotic agents. A rat thrombus model is adopted to evaluate the anti-thrombus activity of the compounds shown in the formula I. Experiment results show that: the compounds shown in the formula I have excellent anti-thrombus activity, and can be used as the antithrombotic agent clinically.

Description

Isoquinoline compound with antithrombotic activity, preparation method and application thereof

technical field

The invention relates to a tricyclic compound, in particular to an isoquinoline compound with antithrombotic activity and a preparation method thereof. The invention also relates to the application of the isoquinoline compound as an antithrombotic agent, which belongs to the field of biomedicine.

Background technique

Vascular embolism is most responsible for the high mortality rate of cardiovascular and cerebrovascular diseases. Thrombosis is the most important etiology of vascular embolism disease. Finding antithrombotic drugs is one of the hot spots in new drug research. Platelet aggregation is related to Ca ion concentration. The concentration of Ca ions in the cytoplasm increases, and platelets are activated and aggregated. The concentration of Ca ions in the cytoplasm decreases, and platelets are inhibited. Most tetrahydroisoquinoline compounds have calcium antagonism and inhibition of platelet aggregation activity. In addition, the benzene ring ultraviolet chromophore in the tetrahydroisoquinoline molecule is convenient for drug metabolism research. Our preliminary studies showed that 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid has a clear antithrombotic effect. Aiming at the fact that 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid has poor fat solubility and water solubility, resulting in poor absorption, we Amino acid residues were introduced into the 2-position N of quinoline-3-carboxylic acid to prepare a series of 3S-N-L-aminoacyl-1,2,3,4-tetrahydroisoquinoline-3-S-carboxylic acid (Peng Shiqi , Zhao Ming, Cui Guohui, Zheng Meiqing, (3S)-N-(L-aminoacyl)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, its synthesis and application as an antithrombotic agent, Application number: 200610164978.7) and introduced amino acid residues on the 3-carboxyl group of 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, prepared a series of 3S-N-1,2,3 , 4-tetrahydroisoquinoline-3-formyl amino acid (Peng Shiqi, Zhao Ming, Cui Guohui, Chen Shenling, N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl] Amino acids, their preparation methods and applications, application number: 200610144236.8), they are all antithrombotic agents.

Introducing L-amino acid into 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid can improve the esterification of 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Solubility and water solubility, but at the same time there is a defect in the metabolism or easy degradation of the introduced L-amino acid in the body, which needs to be overcome.

Contents of the invention

The technical problem to be solved by the present invention at first is to overcome the existing L-amino acid is introduced into 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, there is L-amino acid metabolism in vivo or For the defect of easy degradation, a new isoquinoline compound is provided, and the L-amino acid introduced in the isoquinoline compound is relatively stable and difficult to metabolize in vivo.

The technical problem to be solved in the present invention is realized by the following technical solutions:

Compounds of general formula I below:

Formula I

Wherein, R is selected from CH ₂ CH(CH ₃ ) ₂ , CH(CH ₃ ) ₂ , CH(CH ₃ )CH ₂ CH ₃ , CH ₃ , hydrogen, CH ₂ C ₆ H ₅ , CH ₂ C ₆ H ₄ - OH-p, indol-5-yl-CH ₂ , CH ₂ CO ₂ H, CH ₂ CH ₂ CO ₂ H, CH ₂ CH _{2 CH 2 CH 2} NH ₂ , CH(OH)CH ₃ , CH ₂ CONH _{2 , CH2CH2CONH2 , CH2CH2CH2NHC (} NH) _{NH2 , or CH2CH2SCH3 .}

The present invention has completed the present invention based on and comprehensively considered the following aspects: thrombosis is the most important cause of disease morbidity of vascular embolism; 3S-1,2,3,4-tetrahydroisoline-3-carboxylic acid is Natural substance with anti-platelet aggregation activity; introduction of L-amino acid into 3S-1,2,3,4-tetrahydroisoline-3-carboxylic acid to generate 3S-1,2,3,4-tetrahydroisoline-3 - Formyl amino acid to gain antithrombotic benefit; prevent introduced L-amino acid from being metabolized in vivo to maintain acquired antithrombotic benefit; prevent 3S-1,2,3,4-tetrahydroisoline-3-formyl amino acid from being degraded The most straightforward approach is to generate a third ring between the introduced L-amino acid and the 3S-1,2,3,4-tetrahydroisoline ring.

Another technical problem to be solved by the present invention is to provide a method for preparing the compound of general formula I above.

Another technical problem to be solved by the present invention is achieved through the following technical solutions:

A method for preparing the above-mentioned compound of general formula I, comprising:

(1) Prepare 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid according to conventional methods, and then prepare 3S-2-tert-butoxycarbonyl-6,7-dihydroxyl by tert-butoxycarbonylation -1,2,3,4-Tetrahydroisoquinoline-3-carboxylic acid;

(2) Connect L-amino acid methyl ester or benzyl ester to 3S-2-tert-butoxycarbonyl-6,7-dihydroxyl-1,2,3,4-tetrahydroisoquinoline-3- Preparation of 3S-2-tert-butoxycarbonyl-1,2,3,4-tetrahydroisoquinoline-3-formyl-L-amino acid methyl ester or benzyl ester on the 3-carboxyl group of the carboxylic acid.

(3) 3S-2-tert-butoxycarbonyl-1,2,3,4-tetrahydroisoquinoline-3-formyl-L-amino acid methyl ester or benzyl ester is detached from tert-butoxycarbonyl and then reacted with acetone Ketalization reaction, followed by saponification reaction (in NaOH solution), removal of all protecting groups, yields the compound of general formula I.

In the above preparation method, the 3S-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid in step (1) can be prepared by condensation of L-phenylalanine and formaldehyde.

The L-amino acid methyl ester or benzyl ester described in step (2) is preferably selected from: L-leucine methyl ester, L-valine methyl ester, L-isoleucine methyl ester, L-alanine methyl ester , Glycine methyl ester, L-phenylalanine methyl ester, L-tyrosine methyl ester, L-tryptophan methyl ester, L-aspartic acid dimethyl ester, L-glutamic acid dimethyl ester, L -(N-benzyl)lysine benzyl ester, L-threonine methyl ester, L-asparagine methyl ester, L-glutamine methyl ester L-(NG-nitro)arginine methyl ester or L-methionine methyl ester.

Evaluation of 16 novel 3S-1-(S-carboxy-substituted methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoisocyanurates of the present invention on a rat thrombosis model The antithrombotic activity of quinoline, animal experiment results show that the compound of general formula I of the present invention has excellent antithrombotic activity, and can be used as an antithrombotic agent clinically.

Description of drawings

The synthetic route figure of Fig. 1 general formula I compound of the present invention;

i) concentrated hydrochloric acid, formaldehyde; ii) (Boc) ₂ O, saturated NaHCO ₃ , THF; iii) DCC, HOBt, NMM; iv) 4N HCl/EtOAc, methanol, acetone, triethylamine; v) 2N NaOH/methanol .R=CH ₂ CH(CH ₃ ) ₂ in 3a-5a; R=CH(CH 3 ) ₂ in 3b-5b; R=CH(CH ₃ )CH ₂ CH ₃ in 3c- _5c ; R in 3d-5d =CH ₃ ; R=H in 3e-5e; R=CH ₂ C ₆ H ₅ in 3f-5f; R=CH ₂ C ₆ H ₄ -OH-p in 3g-5g; R=indole in 3h-5h -5-yl- _CH2 ; R _{= CH2CO2Me} in 3i and 4i; R= _CH2CO2H in 5i _; R= _CH2CH2CO2Me in _3j and 4j; R= _CH2 in _5j CH ₂ CO ₂ H; in 3k-5k, R=CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ ; in 31-51, R=CH(OH)CH ₃ ; in 3m-5m, R=CH ₂ CONH ₂ in 3n-5n R=CH ₂ CH ₂ CONH ₂ ; R=CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ in 3o-5o; R=CH ₂ CH ₂ SCH ₃ in 3p-5p.

Detailed ways

In order to further illustrate the present invention, a series of examples are given below. These examples are entirely illustrative, and they are only used to specifically describe the present invention, and should not be construed as limiting the present invention.

The preparation of embodiment 13S-1,2,3,4-tetrahydroisoline-3-carboxylic acid hydrochloride (1)

1.65 g (10 mmol) of L-phenylalanine was dissolved in 90 mL of concentrated hydrochloric acid, and 35 mL of 35% formaldehyde solution was added to the resulting solution. The reaction mixture was reacted at 90° C. for 8-9 hours, and the disappearance of L-phenylalanine was detected by thin-layer chromatography, and the reaction was terminated. The reaction was cooled to room temperature, colorless crystals precipitated, filtered, and the filter cake was washed with acetone to afford 1.92 g (95.4%) of the title compound as a colorless solid. ESI/MS: 178[M+H]+;

The preparation of embodiment 23S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid (2)

4.0 g (18.7 mmol) of 3S-1,2,3,4-tetrahydroisoline-3-carboxylate hydrochloride (1) were dissolved in 40 ml of DMF. To this suspension was added 5.2 g (23.9 mmol) Boc ₂ O with stirring in an ice bath. The pH value of the reaction mixture was adjusted to 10 by adding triethylamine, and the reaction mixture was stirred at room temperature for 48 hours, and the reaction was terminated until the disappearance of the starting material was monitored by thin layer chromatography. Pour the reaction solution into a watch glass and blow it under a fan for about 24 hours until dry. The dried oil was dissolved in 200 mL ethyl acetate, then placed in a 250 mL separatory funnel, and washed with KHSO ₄ (5%) aqueous solution (20 mL×3). Separate the combined ethyl acetate layer, add anhydrous sodium sulfate to a 250 mL Erlenmeyer flask, dry for 0.5 h, and filter under normal pressure. The filtrate was concentrated under reduced pressure until a white solid was precipitated. Filtration under reduced pressure afforded 4.14 g (80%) of the title compound as a colorless solid. ESI/MS: 278 [M+H] ⁺ .

Example 33 Preparation of S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-leucine methyl ester (3a)

Under cooling in an ice bath, place 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid, 0.97g (7.19mmol) N- Hydroxybenzotriazole (HOBt) and 1.37g (7.55mmol) of L-leucine methyl hydrochloride and 70ml of anhydrous tetrahydrofuran (THF) were added dropwise to 1.70g (8.25mmol) of dicyclohexylcarbodiimide (DCC) and 5 mL of anhydrous THF. The pH value of the obtained solution was adjusted to 8 by dropping a solution of NMM and THF. The reaction mixture was stirred at 0°C for 2 hours and then filtered with suction, and the filtrate was concentrated to dryness under reduced pressure with a rotary evaporator. The obtained residue was dissolved in 150 mL of ethyl acetate, placed in a 250 mL separatory funnel, washed successively with saturated aqueous sodium bicarbonate solution 3 times (30 mL each time), washed 3 times with saturated aqueous sodium chloride solution (30 mL each time), 5 Wash with % potassium bisulfate aqueous solution 3 times (30mL each time), wash 3 times with saturated sodium chloride aqueous solution (30mL each time), wash 3 times with saturated sodium bicarbonate aqueous solution (30mL each time), wash 3 times with saturated sodium chloride aqueous solution (30mL each time). The separated ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness with a rotary evaporator under reduced pressure to obtain 2.68 g (92%) of the title compound as a colorless solid. ESI-MS405[M+H] ⁺ ;

Example 43S-1-(1'-methoxycarbonyl-3'-methyl-S-n-butyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b ] The preparation of isoquinoline (4a)

2.00g (4.80mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-leucine methyl ester was placed in a 100mL eggplant-shaped bottle and washed with 8mL ethyl acetate Dissolved, slowly drop into 16mL of 4N HCl/EtOAc in ice bath. The reaction mixture was stirred at 0°C for 1.5 hours and then concentrated to dryness under reduced pressure using a rotary evaporator. After the residue was dissolved in a mixed solvent of 40mL methanol and 40mL acetone and freed, add 20mL acetone, adjust the pH value to 9 with triethylamine, protect with nitrogen, avoid light for 7-10 days, and monitor the disappearance of the raw material point by TLC Stop the reaction. The reaction mixture was concentrated to dryness under reduced pressure with a rotary evaporator (low temperature around 25 degrees Celsius), the residue was dissolved in 200 mL ethyl acetate, the solution was placed in a 250 mL separatory funnel and washed 10 times with 30 mL saturated aqueous sodium chloride solution. The washed ethyl acetate layer was dried over anhydrous sodium sulfate and filtered after half an hour. The filtrate was concentrated to dryness under reduced pressure with a rotary evaporator (low temperature 25 degrees Celsius). The residue was dissolved in 5 mL of methanol, allowed to stand, precipitated, and filtered to obtain 0.76 g (46%) of the title compound as a colorless solid. Mp 186-188°C; ESI-MS (m/e) 345 [M+H] ⁺ ; [α] _D ²⁰ = -58.7 (c = 0.25, chloroform).

Example 51-(1'-carboxy-3'-methyl n-butyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]-isoquinoline (5a )

Dissolve 0.5 g (1.45 mmol) of 1-methoxycarbonyl-methyl-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]-isoquinoline in 10 mL of methanol, Under the condition of ice bath, 10 mL of 2N NaOH solution was slowly added. Thin-layer chromatography monitors the disappearance of raw material spots, and the reaction is terminated. Use 1N HCl to adjust the pH to 7, concentrate under reduced pressure to remove methanol, then use 1N HCl to adjust the pH to 3, and a colorless precipitate precipitates. Filtration, washing the precipitate with distilled water and drying gave 0.42 g (88%) of the title compound as a colorless solid. Mp188-190°C; ESI-MS (m/e) 331 [M+H] ⁺ ; [α] _D ²⁰ = -41.23° (c0.25, methanol)

Example 63 Preparation of S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-valine methyl ester (3b)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3 carboxylic acid and 2.75g (7.55mmol) L-valline hydrochloride Acid methyl ester yielded 2.64 g (95%) of the title compound as a colorless solid. ESI-MS391[M+H] ⁺ .

Example 73S-1-(1'-methoxycarbonyl-2'-methyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b ] Preparation of isoquinoline (4b)

Following the procedure of Example 4 to prepare 4a, 1.44 g (66%) of the title compound was obtained as a colorless solid from 1.87 g (4.8 mmol) of N-Boc-S-isoquinolinoyl-L-valine methyl ester. Mp181-183°C; ESI-MS (m/e) 451 [M+H] ⁺ ; [α] _D ²⁰ =-23.7 (c = 0.25, chloroform)

Example 83S-1-(1'-carboxy-2'-methyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]iso Preparation of quinoline (5b)

According to the operation of Example 5 to prepare 5a, from 0.65g (1.45mmol) 3S-1-(1'-methoxycarbonyl-2'-methyl-S-n-propyl)-2,2-dimethyl-4 -Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.41 g (92%) of the title compound as a colorless solid. Mp174-176°C; ESI-MS (m/e) 317 [M+H] ⁺ ; [α] _D ²⁰ = -8.9 (c = 0.25, methanol)

Example 93 Preparation of S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-isoleucine methyl ester (3c)

According to the operation of Example 3 to prepare 3a, 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.23g (7.34mmol) hydrochloric acid L-iso Leucine methyl ester yielded 2.43 g (95%) of the title compound as a colorless solid. ESI-MS421[M+H] ⁺ . Example 103S-1-(1'-methoxycarbonyl-1'-methyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b ] Preparation of isoquinoline (4c)

According to the operation of preparing 4a in the example, 1.34 g (61%) of the title compound as a colorless solid. Mp192-194°C; ESI-MS (m/e) 465 [M+H] ⁺ ; [α] _D ²⁰ =-58.7 (c = 0.25, chloroform)

Example 113S-1-(1'-carboxy-1'-methyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]iso Preparation of quinoline (5c)

According to the operation of Example 5 to prepare 5a, from 0.67g (1.45mmol) 3S-1-(1'-methoxycarbonyl-1'-methyl-S-n-propyl)-2,2-dimethyl-4 -Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.43 g (93%) of the title compound as a colorless solid. Mp 186-188°C; ESI-MS (m/e) 331 [M+H] ⁺ ; [α] _D ²⁰ =-22.5 (c = 0.25, methanol)

Example 123 Preparation of S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-alanine methyl ester (3d)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.05g (7.55mmol) hydrochloride L-propane Amino acid methyl ester yielded 2.43 g (93%) of the title compound as a colorless solid. ESI-MS362[M+H] ⁺ .

Example 133S-1-(1'-methoxycarbonyl-1'-methyl-S-methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b] Preparation of isoquinoline (4d)

According to the operation of Example 4 to prepare 4a, 1.02 g (51%) of the title compound as a white solid. Mp 201-203 °C; ESI-MS (m/e) 423 [M+H] ⁺ ; [α] _D ²⁰ = -39.0 (c = 0.25, CHCl ₃ ).

Example 143S-1-(1'-carboxy-1'-methyl-S-methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquine Preparation of morphine (5d)

According to the operation of Example 5 to prepare 5a, from 0.61g (1.45mmol) 3S-1-(1'-methoxycarbonyl-1'-methyl-S-methyl)-2,2-dimethyl-4- Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.37 g (92%) of the title compound as a white solid. Mp197-199°C; ESI-MS (m/e) 289 [M+H] ⁺ ; [α] _D ²⁰ =-45.1 (c = 0.25, CH ₃ OH)

The preparation of embodiment 153S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl glycine methyl ester (3e)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 0..99g (7.55mmol) hydrochloric acid L - Glycine methyl ester 2.34 g (93%) of the title compound were obtained as a colorless solid. ESI-MS349[M+H] ⁺ .

Preparation of Example 163S-1-methoxycarbonylmethyl-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquinoline (4e)

According to the operation of Example 4 to prepare 4a, 1.03g (52%) was obtained from 1.67g (4.8mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-formylglycine methyl ester The title compound as a colorless solid. Mp 188-190°C; ESI-MS (m/e) 409 [M+H] ⁺ ; [α] _D ²⁰ = -54.2 (c = 0.25, CHCl ₃ ).

Preparation of Example 173S-1-carboxymethyl-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquinoline (5e)

According to the operation of Example 5 to prepare 5a, from 0.59g (1.45mmol) 3S-1-methoxycarbonylmethyl-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b] Isoquinoline yielded 0.33 g (86%) of the title compound as a colorless solid. Mp 186-188°C; ESI-MS (m/e) 276 [M+H] ⁺ ; [α] _D ²⁰ =-127.6 (c=0.25, CH ₃ OH).

The preparation of embodiment 183S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-phenylalanine methyl ester (3f)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 2.76g (7.55mmol) L-benzene hydrochloride Alanine methyl ester yielded 2.78 g (88%) of the title compound as a colorless solid. ESI-MS439[M+H] ⁺ ;

Example 193S-1-(methoxycarbonyl-phenyl-S-methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquinoline (4f) preparation of

According to the operation of Example 4 to prepare 4a, it was prepared from 2.10g (4.8mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-phenylalanine methyl ester 1.44 g (51%) of the title compound as a colorless solid. Mp191-193°C; ESI-MS (m/e) 499 [M+H] ⁺ ; [α] _D ²⁰ =-96.2 (c0.25, CHCl ₃ ).

Preparation of Example 203S-1-(carboxy-phenyl-S-methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquinoline (5f)

According to the operation of Example 5 to prepare 5a, by 0.72g (1.45mmol) 3S-1-(methoxycarbonyl-phenyl-S-methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazole And[1,5-b]isoquinoline yielded 0.48 g (94%) of the title compound as a white solid. Mp 190-192°C; ESI-MS (m/e) 364 [M+H] ⁺ ; [α] _D ²⁰ =-114.2 (c=0.25, CH ₃ OH).

The preparation of embodiment 213S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-tyrosine methyl ester (3g)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 2.02g (7.55mmol) hydrochloric acid L-phenol Amino acid methyl ester yielded 2.86 g (87%) of the title compound as a colorless solid. ESI-MS455[M+H] ⁺ .

Example 223S-1-(methoxycarbonyl-p-hydroxyphenyl-S-methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquinoline (4g) Preparation

According to the operation of Example 4 to prepare 4a, 1.31 g (53%) of the title compound as a colorless solid. Mp 204-207°C; ESI-MS (m/e) 514 [M+H] ⁺ ; [α] _D ²⁰ = -134.5 (c = 0.25, CHCl ₃ ).

Example 233S-1-(carboxy-p-hydroxyphenyl-S-methyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquinoline (5g ) preparation

According to the operation of Example 5 to prepare 5a, from 0.74g (1.45mmol) 3S-1-(methoxycarbonyl-p-hydroxyphenyl-S-methyl)-2,2-dimethyl-4-oxo- Tetrahydroimidazo[1,5-b]isoquinoline afforded 0.49 g (92%) of the title compound as a colorless solid. Mp 196-198°C; ESI-MS (m/e) 381 [M+H] ⁺ ; [α] _D ²⁰ = -27.1 (c = 0.25, CH ₃ OH).

Example 243 Preparation of S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-tryptophan methyl ester (3h)

According to the operation of Example 3 to prepare 3a, from 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.92g (7.55mmol) hydrochloric acid L-color Amino acid methyl ester yielded 3.12 g (91%) of the title compound as a colorless solid. ESI-MS478[M+H] ⁺ .

Example 253S-1-(1'-methoxycarbonyl-2'-indol-5-yl-S-ethyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1, 5-b] Preparation of isoquinoline (4h)

According to the operation of Example 4 to prepare 4a, 1.46 g (51%) of the title compound as a colorless solid. Mp 174-176°C; ESI-MS (m/e) 538 [M+H] ⁺ ; [α] _D ²⁰ =-127.8 (c=0.25, CHCl ₃ ).

Example 263S-1-(1'-carboxy-2'-indol-5-yl-S-ethyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5- b] Preparation of isoquinoline (5h)

According to the operation of Example 5 to prepare 5a, from 0.78g (1.45mmol) 3S-1-(1'-methoxycarbonyl-2'-indol-5-yl S-ethyl)-2,2-dimethyl -4-Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.46 g (86%) of the title compound as a colorless solid. Mp 175-177°C; ESI-MS (m/e) 404 [M+H] ⁺ ; [α] _D ²⁰ = -112.1 (c = 0.25, CH ₃ OH).

Preparation of Example 273S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-aspartic acid dimethyl ester (3i)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.45g (7.34mmol) hydrochloric acid L-day Dimethyl aspartate yielded 2.88 g (95%) of the title compound as a colorless solid. ESI-MS421[M+H] ⁺ .

Example 283S-1-(1'-methoxycarbonyl-2'-methoxycarbonyl-S-ethyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b ] Preparation of isoquinoline (4i)

According to the operation of preparing 4a in the example, 1.03 g (60%) of the title compound as a colorless solid. Mp 134-136°C; ESI-MS (m/e) 361 [M+H] ⁺ ; [α] _D ²⁰ = -131.1 (c = 0.25, CHCl ₃ ).

Example 293S-1-(1'-carboxy-2'-carboxy-S-ethyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquinoline Preparation of (5i)

According to the operation of Example 5 to prepare 5a, from 0.5g (1.40mmol) 3S-1-(1'-methoxycarbonyl-2'-methoxycarbonyl-S-ethyl)-2,2-dimethyl-4 -Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.40 g (85%) of the title compound as a colorless solid. Mp 126-128°C; ESI-MS (m/e) 333 [M+H] ⁺ ; [α] _D ²⁰ =-120.4 (c=0.25, CH ₃ OH).

The preparation of embodiment 303S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-glutamic acid dimethyl ester (3j)

According to the operation of Example 3 to prepare 3i, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.60g (7.55mmol) hydrochloride L-glucose Amino acid dimethyl ester yielded 2.98 g (95%) of the title compound as a colorless solid. ESI-MS435[M+H] ⁺ .

Example 313S-1-(1'-methoxycarbonyl-2'-methoxycarbonyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5- b] Preparation of isoquinoline (4j)

According to the operation of Example 4 to prepare 4i, it was prepared from 2.08g (4.8mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-glutamic acid dimethyl ester 1.34 g (56%) of the title compound as a colorless solid. Mp 181-182°C; ESI-MS (m/e) 495 [M+H] ⁺ ; [α] _D ²⁰ =-140.2 (c=0.25, CHCl ₃ ).

Example 323S-1-(1'-carboxy-2'-carboxy-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquine Preparation of morphine (5j)

According to the operation of Example 5 to prepare 5i, from 0.72g (1.45mmol) 3S-1-(1'-methoxycarbonyl-2'-methoxycarbonyl-S-n-propyl)-2,2-dimethyl- 4-Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.42 g (87%) of the title compound as a colorless solid. Mp 172-175°C; ESI-MS (m/e) 347 [M+H] ⁺ ; [α] _D ²⁰ =-74.2 (c=0.25, CH ₃ OH).

The preparation of embodiment 333S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-(N-benzyl)lysine benzyl ester (3k)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 2.50g (7.55mmol) LN-benzyl hydrochloride Benzyl lysine benzyl ester yielded 3.41 g (85%) of the title compound as a colorless solid. ESI-MS554[M+H] ⁺ .

Example 343S-1-(1'-benzyloxycarbonyl-6'-benzyloxycarbonyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5- b] Preparation of isoquinoline (4k)

According to the operation of Example 4 to prepare 4a, by 2.66g (4.8mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-(N-benzyl)lysine Benzyl acid ester yielded 1.24 g (42%) of the title compound as a colorless solid. Mp 136-138°C; ESI-MS (m/e) 614 [M+H] ⁺ ; [α] _D ²⁰ =-114.3 (c=0.25, CHCl ₃ ).

Example 353S-1-(1'-carboxy-6'-amino-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquine Preparation of morphine (5k)

After the operation of preparing 5a according to Example 5, dissolve the insoluble matter in 30ml methanol, react under hydrogen/palladium carbon conditions, react for 1 day, until the raw material point disappears, filter, and spin dry at a low temperature (25 degrees Celsius) to obtain the final product, obtained by 0.89 g (1.45mmol) 3S-1-(1'-benzyloxycarbonyl-6'-benzyloxycarbonyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1 ,5-b] Isoquinoline yielded 0.42 g (87%) of the title compound as a colorless solid. Mp 141-143°C; ESI-MS (m/e) 346 [M+H] ⁺ ; [α] _D ²⁰ =-107.1 (c=0.25, CH ₃ OH).

Example 363 Preparation of S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-threonine methyl ester (31)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.28g (7.55mmol) L-threohydrochloride Amino acid methyl ester yielded 2.31 g (82%) of the title compound as a colorless solid. ESI-MS393[M+H] ⁺ .

Example 373S-1-(1'-methoxycarbonyl-2'-hydroxyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b] Preparation of isoquinoline (41)

According to the operation of Example 4 to prepare 4a, 0.94 g (42%) of the title compound as a colorless solid. Mp 201-203°C; ESI-MS (m/e) 453 [M+H] ⁺ ; [α] _D ²⁰ =-59.2 (c=0.25, CHCl ₃ ).

Example 383S-1-(1'-carboxy-2'-hydroxyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquine Preparation of morphine (51)

According to the operation of Example 5 to prepare 5a, from 0.66g (1.45mmol) 3S-1-(1'-methoxycarbonyl-2'-hydroxyl-S-n-propyl)-2,2-dimethyl-4- Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.38 g (85%) of the title compound as a white solid. Mp 196-198°C; ESI-MS (m/e) 319 [M+H] ⁺ ; [α] _D ²⁰ = -29.2 (c = 0.25, CH ₃ OH).

The preparation of embodiment 393S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-asparagine methyl ester (3m)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.38g (7.55mmol) hydrochloric acid L-day Paragine methyl ester yielded 2.13 g (72%) of the title compound as a colorless solid. ESI-MS406[M+H] ⁺ .

Example 403S-1-(1'-methoxycarbonyl-2'-aminocarbonyl-S-ethyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b] Preparation of isoquinoline (4m)

According to the operation of Example 4 to prepare 4a, 0.96 g (42%) of the title compound as a colorless solid. Mp 183-185°C; ESI-MS (m/e) 466 [M+H] ⁺ ; [α] _D ²⁰ =-109.2 (c=0.25, CHCl ₃ ).

Example 413S-1-(1'-carboxy-2'-aminocarbonyl-S-ethyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]isoquine Preparation of morphine (5m)

According to the operation of Example 5 to prepare 5a, from 0.67g (1.45mmol) 3S-1-(1'-methoxycarbonyl-2'-aminocarbonyl-S-ethyl)-2,2-dimethyl-4- Oxo-tetrahydroimidazo[1,5-b]isoquinoline afforded 0.41 g (88%) of the title compound as a colorless solid. Mp 192-194°C; ESI-MS (m/e) 332 [M+H] ⁺ ; [α] _D ²⁰ =-169.1 (c=0.25, CH ₃ OH).

The preparation of embodiment 423S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-glutamine methyl ester (3n)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.48g (7.55mmol) hydrochloride L-glucose Aminoamidomethyl ester yielded 2.53 g (84%) of the title compound as a colorless solid. ESI-MS420[M+H] ⁺ .

Example 433S-1-(1'-methoxycarbonyl-2'-aminocarbonyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b ] Preparation of isoquinoline (4n)

According to the operation of Example 4 to prepare 4a, 0.98 g (42%) of the title compound as a colorless solid. Mp 159-161 °C; ESI-MS (m/e) 480 [M+H] ⁺ ; [α] _D ²⁰ = -140.2 (c = 0.25, CHCl ₃ ).

Example 443S-1-(1'-carboxy-2'-aminocarbonyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]iso Preparation of quinoline (5n)

According to the operation of Example 5 to prepare 5a, from 0.69g (1.45mmol) 3S-1-(1'-methoxycarbonyl-2'-aminocarbonyl-S-n-propyl)-2,2-dimethyl-4 -Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.43 g (89%) of the title compound as a colorless solid. Mp 156-158°C; ESI-MS (m/e) 346 [M+H] ⁺ ; [α] _D ²⁰ =-24.3 (c=0.25, CH ₃ OH).

The preparation of embodiment 453S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-( ^NG -nitro)arginine methyl ester (3o)

According to the operation of Example 3 to prepare 3a, 2.04g (7.55mmol) hydrochloric acid L-( ( ^NG -nitro)arginine methyl ester yielded 3.12 g (87%) of the title compound as a colorless solid. ESI-MS492[M+H] ⁺ .

Example 463S-1-(1'-methoxycarbonyl-4'-nitroguanidino-S-n-butyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5 -b] Preparation of isoquinoline (4o)

According to the operation of Example 4 to prepare 4a, by 2.36g (4.8mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-( ^NG -nitro) Amino acid methyl ester yielded 1.14 g (43%) of the title compound as a colorless solid. Mp 186-188°C; ESI-MS (m/e) 552 [M+H] ⁺ ; [α] _D ²⁰ = -147.1 (c = 0.25, CHCl ₃ ).

Example 473S-1-(1'-carboxy-4'-guanidino-S-n-butyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b]iso Preparation of quinoline (5o)

After preparing 5a according to Example 5, dissolve the insoluble matter in 30ml of methanol, react under hydrogen/palladium carbon conditions, react for 1 day, until the raw material point disappears, filter, and spin dry at low temperature (25 degrees Celsius). From 0.80g (1.45mmol) 3S-1-(1'-methoxycarbonyl-4'-nitroguanidino-S-n-butyl)-2,2-dimethyl-4-oxo-tetrahydroimidazole And[1,5-b]isoquinoline yielded 0.42 g (88%) of the title compound as a colorless solid. Mp 181-183°C; ESI-MS (m/e) 373 [M+H] ⁺ ; [α] _D ²⁰ =-35.0 (c=0.25, CH ₃ OH).

The preparation of embodiment 483S-N-Boc-1,2,3,4-tetrahydroisoline-3-formyl-L-methionine methyl ester (3p)

According to the operation of Example 3 to prepare 3a, by 2.00g (7.22mmol) 3S-N-Boc-1,2,3,4-tetrahydroisoline-3-carboxylic acid and 1.50g (7.55mmol) L-methionine hydrochloride The methyl ester yielded 2.35 g (77%) of the title compound as a colorless solid. ESI-MS423[M+H] ⁺ .

Example 493S-1-(1'-methoxycarbonyl-3'-methylthio-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5- b] Preparation of isoquinoline (4p)

According to the operation of Example 4 to prepare 4a, 0.86 g ( 37%) the title compound as a colorless solid. Mp 174-176°C; ESI-MS (m/e) 483 [M+H] ⁺ ; [α] _D ²⁰ = -89.1 (c = 0.25, CHCl ₃ ).

Example 503S-1-(1'-carboxy-3'-methylthio-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5-b] Preparation of isoquinoline (5p)

According to the operation of Example 5 to prepare 5a, from 0.70g (1.45mmol) 3S-1-(1'-methoxycarbonyl-3'-methylthio-S-n-propyl)-2,2-dimethyl- 4-Oxo-tetrahydroimidazo[1,5-b]isoquinoline yielded 0.44 g (90%) of the title compound as a colorless solid. Mp 182-184°C; ESI-MS (m/e) 349 [M+H] ⁺ ; [α] _D ²⁰ =-61.7 (c=0.25, CH ₃ OH).

Test example 1 The antithrombotic activity of the compound of the present invention (5a-p) is measured

1) Experimental animals: SD male rats, weighing 200-260g (purchased from the Experimental Animal Department of Beijing Medical University, license number is Yidongzi No. 01-3056)

2) Experimental materials: polyethylene tubes (both 1.6 mm and 1.3 mm in outer diameter), heparin (50 IU/ml), and sodium pentobarbital (5%).

3) Measurement method

Male SD rats (body weight 200g to 260g) were anesthetized by intraperitoneal injection of pentobarbital sodium solution, and the right common carotid artery and left external jugular vein were separated, and a previously weighed 6cm long silk thread was placed in the middle of the polyethylene tube. Fill the polyethylene tube with heparin normal saline (50IU/kg), insert one end into the left external jugular vein, inject normal saline (3ml/kg), aspirin normal saline solution (20mg/kg), and the embodiment of the present invention respectively The prepared saline solution of compound 5a-p (10nmol/kg) was slowly injected into the polyethylene tube from the other end. At this time, the original heparin in the tube had been pushed into the left external jugular vein, and most of the tested compounds were in the tube. solution, and then insert the injection end into the right common carotid artery. The blood flowed from the right common carotid artery to the left external jugular vein through the polyethylene tube. After 15 minutes, the blood flow was interrupted. The silk thread was taken out and weighed. The wet weight of the thrombus was subtracted from the total weight of the silk thread. The average value and standard of the thrombus wet weight in each group were calculated Difference (X ± DS), and t test.

4) Results: See Table 1.

Table 15a-p Antithrombotic activity

n=10, the dose of aspirin=20mg/kg; The dose of isoquinoline carboxylic acid=5μmol/kg; The dose of 5a-p=10nmol/kg; a) p<0.01 compared with normal saline group; b) compared with normal saline Group ratio p<0.05;.

Test Example 2 Compound 3S-1-(1'-carboxy-1'-methyl-S-n-propyl)-2,2-dimethyl-4-oxo-tetrahydroimidazo[1,5 -b] the dose-effect relationship of isoquinoline (5c)

1) Experimental animals: SD male rats, weighing 200-260g (purchased from the Experimental Animal Department of Beijing Medical University, license number is Yidongzi No. 01-3056)

2) Experimental materials: polyethylene tubes (both 1.6 mm and 1.3 mm in outer diameter), heparin (50 IU/ml), and sodium pentobarbital (5%).

3) Measurement method

Male SD rats (body weight 200g to 260g) were anesthetized by intraperitoneal injection of pentobarbital sodium solution, and the right common carotid artery and left external jugular vein were separated, and a previously weighed 6cm long silk thread was placed in the middle of the polyethylene tube. Fill the polyethylene tube with heparin saline (50IU/kg), insert one end into the left external jugular vein, and slowly inject 5c (100nmol/kg, 10nmol/kg, 1nmol/kg) normal saline solution into the polyethylene tube from the other end with a syringe At this time, the original heparin in the tube has been pushed into the left external jugular vein, and most of the tube is the test compound solution, and then the drug injection end is inserted into the right common carotid artery. The blood flowed from the right common carotid artery to the left external jugular vein through the polyethylene tube. After 15 minutes, the blood flow was interrupted. The silk thread was taken out and weighed. The wet weight of the thrombus was subtracted from the total weight of the silk thread. The average value and standard of the thrombus wet weight in each group were calculated Difference (X ± DS), and t test.

4) Results: See Table 2.

Table 25c Antithrombotic activity at different concentrations

dose 100nmol/kg 10nmol/kg 1mmol/kg Thrombus weight (X±SDmg) 12.02±2.47^a 12.99±2.05^a 20.67±2.56

n=10; a) p<0.01 compared with 1nmol/kg group

Claims (5)

1. compound of Formula I:

General formula I

Wherein, R is selected from CH ₂CH (CH ₃) ₂, CH (CH ₃) ₂, CH (CH ₃) CH ₂CH ₃, CH ₃, hydrogen, CH ₂C ₆H ₅, CH ₂C ₆H ₄-OH-p, indol-3-yl-CH ₂, CH ₂CO ₂H, CH ₂CH ₂CO ₂H, CH ₂CH ₂CH ₂CH ₂NH ₂, CH (OH) CH ₃, CH ₂CONH ₂, CH ₂CH ₂CONH ₂, CH ₂CH ₂CH ₂NHC (=NH) NH ₂Or CH ₂CH ₂SCH ₃

2. method for preparing the described compound of Formula I of claim 1 comprises:

(1) prepare 3S-1 according to ordinary method, 2,3,4-tetrahydroisoquinoline-3-carboxylic acid, t-butoxycarbonylating prepares 3S-2-tertbutyloxycarbonyl-1,2,3 then, 4-tetrahydroisoquinoline-3-carboxylic acid;

(2) earlier L-amino acid methyl ester or benzyl ester are connected to 3S-2-tertbutyloxycarbonyl-1,2,3 according to ordinary method, preparation 3S-2-tertbutyloxycarbonyl-1 on 3 carboxyls of 4-tetrahydroisoquinoline-3-carboxylic acid, 2,3,4-tetrahydroisoquinoline-3-formyl-L-amino acid methyl ester or benzyl ester; Described L-amino acid methyl ester or benzyl ester are selected from L-leucine methyl esters, L-valine methyl ester, L-Isoleucine methyl esters, L-alanine methyl ester, glycine methyl ester, L-phenylalanine methyl ester, L-L-Tyrosine methyl ester, L-tryptophan methyl ester, L-aspartic acid dimethyl ester, L-glutamic acid dimethyl ester, L-(N-benzyl) Methionin benzyl ester, L-Threonine methyl esters, altheine methyl esters, L-glutaminate methyl esters, L-(N ^G-nitro) arginine methyl esters or L-methyl methionine;

(3) with 3S-2-tertbutyloxycarbonyl-1,2,3,4-tetrahydroisoquinoline-3-formyl-L-amino acid methyl ester or benzyl ester slough behind the tertbutyloxycarbonyl with acetone carry out ketal reaction, again carry out saponification reaction, slough all protecting groups, obtain compound of Formula I.

3. according to the method for claim 2, it is characterized in that: 3S-1 described in the step (1), 2,3,4-tetrahydroisoquinoline-3-carboxylic acid is prepared by L-phenylalanine and formaldehyde condensation.

4. a treatment antithrombotic pharmaceutical composition is made up of the described compound of Formula I of the claim 1 of significant quantity and pharmaceutically acceptable carrier or auxiliary material.

5. the described compound of Formula I of claim 1 is in the purposes of preparation in the antithrombotic reagent.

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