2-Trp-AA-tetrahydrocarboline-3-carboxylic acid compounds selectively inhibiting ADP and preparation and application thereof

Abstract

The present invention discloses a 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compound with the selective inhibition of adenosine diphosphate (ADP). The structural formula of the compound is as follows, wherein the amino acid represented by AA is selected from L-Pro residue, L-Tyr residue, L-Gln residue and L-His residue, respectively. The present invention further discloses a preparation method of the compound and its application in the treatment of arterial thrombotic diseases. Experiments have shown that the ADP selective inhibitor of the present invention has a good anti-arterial thrombotic effect. Therefore, it is proposed that the present invention provides an effective technical means for anti-arterial thrombosis. #imgabs0#

Description

Technical Field

[0001] The present invention relates to 2-Trp-AA-tetrahydrocarboline-3-carboxylic acid compounds which selectively inhibit adenosine diphosphate (ADP), and to their preparation methods and applications in treating arterial thrombotic diseases. The present invention belongs to the field of biomedicine. Background Art

[0002] Arterial embolism has become one of the diseases with high morbidity and mortality. Arterial thrombosis is responsible for transient ischemic attack, acute coronary syndrome, myocardial infarction and atrial fibrillation. Among patients with atrial fibrillation, 18%-47% have coronary artery disease, and about 20% of patients with atrial fibrillation with coronary artery disease receive percutaneous coronary intervention. Arterial thrombosis is also responsible for arterial thrombosis and unstable angina after artificial heart valves, arteriovenous fistulas and other surgeries. For example, after liver transplantation, patients are at risk of hepatic artery thrombosis. In addition, patients with antiphospholipid syndrome are also at risk of arterial thrombosis. Arterial thrombosis is associated with platelet aggregation. Inducers that usually induce platelet aggregation include platelet activating factor (PAF), adenosine diphosphate (ADP), thrombin (TH) and arachidonic acid (AA). The discovery of selective inhibitors of adenosine diphosphate is an important direction for the research of anti-arterial thrombotic drugs. In the study, the inventors screened out the following structure 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid (wherein AA is L-Pro residue, L-Tyr residue, L-Gln residue and L-His) which selectively inhibits ADP-induced platelet aggregation.

[0003]

[0004] Four 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acids showed excellent anti-arterial thrombotic activity in the rat silk thread arterial thrombosis model. Based on these findings, the inventors proposed the present invention. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide four types of 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compounds with selective inhibition of ADP. Experiments have shown that the 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compounds prepared by the present invention selectively inhibit adenosine diphosphate (ADP) and have excellent anti-arterial thrombotic effects. In order to achieve the above technical effects, the present invention adopts the following four technical means.

[0006] The first technical means is to confirm the 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compound of the present invention, whose structural formula is as follows:

[0007]

[0008] The amino acid represented by AA in the formula is selected from: L-Pro residue, L-Tyr residue, L-Gln residue or L-His.

[0009] The second technical means is to propose a method for preparing the 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid, which comprises the following steps:

[0010] 1) preparing 3S-tetrahydro-β-carboline-3-carboxylic acid;

[0011] 2) preparing Boc-Trp-AA, wherein AA is L-Pro residue, L-Tyr residue, L-Gln residue and L-His;

[0012] 3) preparing 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid, wherein AA is L-Pro residue, L-Tyr residue, L-Gln residue and L-His;

[0013] 4) preparing 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid, wherein AA is L-Pro residue, L-Tyr residue, L-Gln residue and L-His.

[0014] The third technical means is to confirm the selective inhibitory effect of the 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compounds on adenosine diphosphate (ADP).

[0015] The fourth technical means is to confirm the selective inhibitory effect of the 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compounds on ADP-induced platelet aggregation.

[0016] The fifth technical means is to confirm the excellent inhibitory effect of the 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid on arterial thrombosis.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The present invention modifies carboline on the basis of its structure to prepare a class of 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acids with the above structure. Experiments show that the compounds show good activity in inhibiting adenosine diphosphate (ADP) and inhibiting arterial thrombosis.

[0019] The invention provides a new therapeutic drug for the treatment of arterial thrombosis, and enriches people's drug selection for the treatment of arterial thrombosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The synthetic route of 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid is as follows: i) CH2O, H2SO4; ii) DCC, HOBt, N-methylmorpholine, Boc-Trp-AA; iii) 4N hydrogen chloride solution in ethyl acetate. DETAILED DESCRIPTION

[0021] In order to further illustrate the present invention, a series of embodiments are given below. These embodiments are purely illustrative and are only used to specifically describe the present invention and should not be construed as limiting the present invention.

[0022] Example 1 Preparation of 3S-tetrahydro-β-carboline-3-carboxylic acid (1)

[0023] Slowly add 0.2 mL of concentrated sulfuric acid to 400 mL of water. Add 5.0 g (24.5 mmol) of L-Trp to the obtained dilute sulfuric acid aqueous solution, and ultrasonically oscillate until L-Trp is completely dissolved. Add 10 mL of 35% formaldehyde aqueous solution to the obtained solution. The reaction mixture is stirred at room temperature for 6 hours, and the reaction is terminated when L-Trp disappears as monitored by thin layer chromatography (ethyl acetate / ether, 20 / 1). Slowly add concentrated ammonia water to the reaction solution, adjust to pH 6, and let stand for 30 minutes. Filter out the generated precipitate and wash with water. Spread the filtered colorless solid on a petri dish and dry in air to obtain 5.05 g (95%) of the title compound as a colorless solid. ESI-MS (m / e): 217 [M+H] + .

[0024] Example 2 Preparation of Boc-Trp AA

[0025] Boc-Trp-Pro-O-CH3, Boc-Trp-Tyr-O-CH3, Boc-Trp-Gln-O-CH3 and Boc-Trp-His-O-CH3 were prepared from Boc-Trp and Pro-O-CH3, Tyr-O-CH3, Gln-O-CH3 and His-O-CH3 by conventional DCC method. Boc-Trp-Pro-O-CH3, Boc-Trp-Tyr-O-CH3, Boc-Trp-Gln-O-CH3 and Boc-Trp-His-O-CH3 were saponified in 2N sodium hydroxide solution at 0℃ to obtain Boc-Trp-Pro, ESI-MS: 402[M+H] + ;Boc-Trp-Tyr,ESI-MS:468[M+H] + ;Boc-Trp-Gln,ESI-MS:433[M+H] +and Boc-Trp-His, ESI-MS: 442[M+H] + Their yields all reached 91%, their purities all exceeded 98%, and they were all known compounds, so they were directly used in the subsequent synthetic reactions without being tested.

[0026] Example 3 Preparation of 3S-[2-(Boc-Trp-Pro)]-tetrahydro-β-carboline-3-carboxylic acid (2a)

[0027] 2.0 g (14.8 mmol) N-hydroxybenzotriazole (HOBt) and 3.1 g (14.8 mmol) dicyclohexylcarbodiimide (DCC) were added to a solution of 3.03 g (14 mmol) 3S-tetrahydro-β-carboline-3-carboxylic acid, 5.6 g (14 mmol) Boc-Trp-Pro and 150 mL anhydrous tetrahydrofuran at 0°C. The reaction mixture was stirred at 0°C for 30 minutes. Then the pH was adjusted to 8 with N-methylmorpholine (NMM). The reaction mixture was stirred at 0°C for 6 hours. The reaction was terminated when 3S-tetrahydro-β-carboline-3-carboxylic acid disappeared as monitored by thin layer chromatography (ethyl acetate / methanol, 20 / 1). The reaction mixture was filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was dissolved in 200 mL ethyl acetate. The obtained solution was washed with 5% sodium bicarbonate aqueous solution (30 mL × 3), saturated sodium chloride aqueous solution (30 mL × 3), 5% hydrochloric acid aqueous solution (30 mL × 3), and saturated sodium chloride aqueous solution (30 mL × 3). The ethyl acetate layer was separated, dried over anhydrous sodium sulfate for 12 hours, filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain 7.55 g (90%) of the title compound as a colorless powder. ESI-MS: 600 [M+H] + ; 1H NMR (300MHz, DMSO-d6): δ / ppm=12.891(s,1H),11.675(s,1H),10.791(s,1H),7.582(d,J=7.6Hz,1H),7.352(d,J=7.7Hz,1H),7.3 95(s,1H),7.332(d,J=7.9Hz,1H),7.213(d,J=7.9Hz,1H),7.211(s,1H),7.064(t,J=7.2Hz,1H),6.981(t,J=7.2Hz,1H),6.933(t, J=7.2Hz,1H),6.912(t,J=7.2Hz,1H),4.925(t,J=5.6Hz,1H),4.856(t,J1=6.1Hz,1H),4.553(s,2H),4.402(d,J=5.6Hz,1H),3.5 22(t,J=5.7Hz,2H),3.312(d,J=5.6Hz,2H),2.332(m,J=5.6Hz,2H),2.023(m,J=5.6Hz,2H),3.042(d,J=6.1Hz,2H),1.424(s,9H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=171.91,170.72,169.35,160.22,136.53,136.22,130.43,127.45,127.31,123.11,121.78,121.72,119.86,119.8 3,118.85,118.75,111.16,111.13,109.87,104.77,79.65,67.83,66.03 ,57.33,49.53,38.15,29.63,28.45,28.44,28.43,28.12,24.13,22.65.

[0028] Example 4 Preparation of 3S-[2-(Trp-Pro)]-tetrahydro-β-carboline-3-carboxylic acid (3a)

[0029] Dissolve 5.6 g (9.3 mmol) of 3S-[2-(Boc-Trp-Pro)]-tetrahydro-β-carboline-3-carboxylic acid in 50 mL of ethyl acetate solution (4N) of hydrogen chloride, and stir the reaction mixture at room temperature for 60 minutes. The reaction was terminated when 3S-[2-(Boc-Trp-Pro)]-tetrahydro-β-carboline-3-carboxylic acid disappeared as monitored by thin layer chromatography (ethyl acetate / methanol, 20 / 1). The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in 50 mL of ethyl acetate and concentrated again under reduced pressure. This operation was repeated 5 times to obtain 4.55 g (98%) of the title compound as a colorless powder. FT-ICR-MS: 500.2298 [M+H] + ; 1 H NMR (300MHz, DMSO-d6): δ / ppm=12.892(s,1H),11.673(s,1H),10.793(s,1H),8.861(s,2H),7.583(d,J=7.6Hz,1H),7.3 51(d,J=7.7Hz,1H),7.333(d,J=7.9Hz,1H),7.212(d,J=7.9Hz,1H),7.201(s,1H),7.065(t,J=7.2Hz,1H),6.982(t,J=7. 2Hz,1H),6.932(t,J=7.2Hz,1H),6.911(t,J=7.2Hz,1H),4.855(t,J1=6.1Hz,1H),4.551(s,2H),4.402(t,J=6.9Hz,1H) ,3.952(t,J=5.6Hz,1H),3.512(t,J=6.9Hz,2H),3.312(d,J=5.6Hz,2H),2.331(m,J=6.9Hz,2H),2.022(m,J=6.9Hz,2H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=173.31,171.72,169.36,136.52,136.23,130.44,127.43,127.32,123.01,121.77,121.73,119 .85,119.84,118.86,118.76,111.15,111.14,107.17,104.89,67.85,66.05,52.63,49.52,38.13,29.83,27.72,24.15,22.64.

[0030] Example 5 Preparation of 3S-[2-(Boc-Trp-Tyr)]-tetrahydro-β-carboline-3-carboxylic acid (2b)

[0031] Using the procedure of Example 3, 8.4 g (90%) of the title compound were obtained from 3.03 g (14 mmol) of 3S-tetrahydro-β-carboline-3-carboxylic acid and 6.05 g (14 mmol) of Boc-Trp-Tyr as a colorless powder. ESI-MS: 666 [M+H] + ; 1 H NMR (300MHz, DMSO-d6): δ / ppm=12.891(s,1H),11.671(s,1H),10.786(s,1H),9.086(s,1H),8.321(s,1H),7.582(d,J=7.6Hz,1H),7.392( s,1H),7.355(d,J=7.9Hz,1H),7.333(d,J=7.9Hz,1H),7.211(s,1H),7.213(d,J=7.9Hz,1H),7.061(t,J=7.2Hz,1H),6.981(t,J=7.2Hz,1 H), 6.962 (t, J = 7.2Hz, 2H), 6.933 (t, J = 7.2Hz, 1H), 6.912 (t, J = 7.2Hz, 1H), 6.681 (t, J = 7.2Hz, 2H), 4.924 (t, J = 5.6Hz, 1H), 4.922 (t, J = 5. 6Hz,1H),4.851(t,J1=6.1Hz,1H),4.454(s,2H),3.452(d,J1=5.6Hz,2H),3.314(d,J1=5.6Hz,2H),3.045(d,J1=6.1Hz,2H),1.421(s,9H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=171.95,171.72,170.06,155.95,155.75,136.52, 136.23,130.40,130.21,130.20,129.22,127.44,127.32,123.15,121.77,121.71 ,119.84,119.82,118.81,118.80,115.82,115.81,111.15,111.13,109.72,104.95,79.55,67.88,59.53,55.13,38.03,37.75,28.41,28.40,28.39,27.85,22.63.

[0032] Example 6 Preparation of 3S-[2-(Trp-Tyr)]-tetrahydro-β-carboline-3-carboxylic acid (3b)

[0033] 4.26 g (93%) of the title compound was obtained from 5.4 g (8.1 mmol) of 3S-[2-(Boc-Trp-Tyr)]-tetrahydro-β-carboline-3-carboxylic acid as a colorless powder using the procedure of Example 4. FT-ICR-MS: 566.2403; 1 H NMR (300MHz, DMSO-d6): δ / ppm=12.892(s,1H),11.672(s,1H),10.782(s,1H),9.083(s,1H),8.861(s,2H),8.322(s,1H),7.581(d,J=7.6Hz,1H),7 .353(d,J=7.9Hz,1H),7.332(d,J=7.9Hz,1H),7.212(s,1H),7.212(d,J= 7.9Hz,1H),7.063(t,J=7.2Hz,1H),6.982(t,J=7.2Hz,1H),6.962(d,J=7. 2Hz,1H),6.961(t,J=7.2Hz,1H),6.931(t,J=7.2Hz,1H),6.912(t,J=7.2 Hz,1H),6.682(t,J=7.2Hz,1H),6.681(t,J=7.2Hz,1H),4.921(t,J=5.6H z,1H),4.852(t,J1=6.1Hz,1H),4.456(s,2H),3.952(t,J1=5.6Hz,1H),3 .443(d,J1=5.6Hz,2H),3.313(d,J1=5.6Hz,2H),3.043(d,J1=6.1Hz,2H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=171.93,171.73,170.05,155.74,136.51,136.22,130.41,130.22,130.21,129.23,127.43,127.31,123.13,121. 75,121.72,119.85,119.83,118.83,118.81,115.83,115.82,111.14,11 1.13,107.12,104.93,67.85,56.73,55.13,38.04,37.74,27.45,22.62.

[0034] Example 7 Preparation of 3S-[2-(Boc-Trp-Gln)]-tetrahydro-β-carboline-3-carboxylic acid (2c)

[0035] Using the procedure of Example 3, 7.94 g (90%) of the title compound was obtained from 3.03 g (14 mmol) of 3S-tetrahydro-β-carboline-3-carboxylic acid and 5.84 g (14 mmol) of Boc-Trp-Gln as a colorless powder. ESI-MS: 631 [M+H] + ; 1 H NMR (300MHz, DMSO-d6): δ / ppm=12.892(s,1H),11.673(s,1H),10.789(s,1H),8.321(s,1H),7.582(d,J=7.6Hz,1H),7.391(s,1H),7 .351(d,J=7.9Hz,1H),7.333(d,J=7.9Hz,1H),7.203(s,1H),7.213(d,J=7.9Hz,1H),7.062(t,J=7.2Hz,1H),7.033(s,2H),6.981(t, J=7.2Hz,1H),6.931(t,J=7.2Hz,1H),6.912(t,J=7.2Hz,1H),4.853(t,J=5.6Hz,1H),4.922(t,J=5.6Hz,1H),4.440(t,J1=6.1Hz,1 H),4.555(s,2H),3.314(d,J1=5.6Hz,2H),3.045(d,J1=6.1Hz,2H),2.071(m,J1=6.1Hz,2H),2.051(t,J1=6.1Hz,2H),1.421(s,9H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=173.56,171.90,171.66,169.32,155.97,136.52,136.21,130.41,127.41,127.31,123.13,121.71,121.69,119.8 4,119.82,118.81,118.80,111.14,111.13,109.72,104.93,79.55,67.8 7,59.55,54.62,38.03,32.61,28.41,28.40,28.39,27.87,27.85,22.63.

[0036] Example 8 Preparation of 3S-[2-(Trp-Gln)]-tetrahydro-β-carboline-3-carboxylic acid (3c)

[0037] 4.2 g (90%) of the title compound was obtained as a colorless powder from 5.6 g (8.89 mmol) of 3S-[2-(Boc-Trp-Gln)]-tetrahydro-β-carboline-3-carboxylic acid using the procedure of Example 4. FT-ICR-MS: 531.2356 [M+H] + ; 1 H NMR (300MHz, DMSO-d6): δ / ppm=12.891(s,1H),11.671(s,1H),10.787(s,1H),8.861(s,2H),8.321(s,1H),7.581(d,J=7.6Hz,1H ),7.352(d,J=7.9Hz,1H),7.332(d,J=7.9Hz,1H),7.201(s,1H),7.212(d,J=7.9Hz,1H),7.061(t,J=7.2Hz,1H),7.031(s,2H),6. 982(t,J=7.2Hz,1H),6.932(t,J=7.2Hz,1H),6.911(t,J=7.2Hz,1H),4.852(t,J=5.6Hz,1H),4.441(t,J1=6.1Hz,1H),4.553(s, 2H),3.951(t,J1=5.6Hz,1H),3.315(d,J1=5.6Hz,2H),3.042(d,J=5.6Hz,2H),2.072(m,J1=6.1Hz,2H),2.052(t,J1=6.1Hz,2H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=173.57,171.91,171.69,169.33,136.53,136.22,130.43,127.42,127.32,123.12,121.73,121. 72,119.83,119.81,118.82,118.81,111.13,111.12,107.13,104.93,67.85,56.72,54.62,38.05,32.63,27.86,27.45,22.64.

[0038] Example 9 Preparation of 3S-[2-(Boc-Trp-His)]-tetrahydro-β-carboline-3-carboxylic acid (2d)

[0039] Using the procedure of Example 3, 8.1 g (90%) of the title compound were obtained from 3.03 g (14 mmol) of 3S-tetrahydro-β-carboline-3-carboxylic acid and 5.25 g (14 mmol) of Boc-Trp-His as a colorless powder. ESI-MS: 640 [M+H] + ;1 H NMR (300MHz, DMSO-d6): δ / ppm=13.091(s,1H),12.891(s,1H),11.671(s,1H),10.788(s,1H),8.732(s,1H),8.322(s,1H),7.662(s,1 H),7.582(d,J=7.6Hz,1H),7.391(s,1H),7.351(d,J=7.9Hz,1H),7.332(d,J=7.9Hz,1H),7.203(s,1H),7.213(d,J=7.9Hz,1H),7.06 1(t,J=7.2Hz,1H),6.981(t,J=7.2Hz,1H),6.931(t,J=7.2Hz,1H),6.912(t,J=7.2Hz,1H),4.924(t,J=5.6Hz,1H),4.922(t,J=5.6Hz ,1H),4.852(t,J1=6.1Hz,1H),4.454(s,2H),3.314(d,J1=5.6Hz,2H),3.181(t,J=5.6Hz,2H),3.043(d,J1=6.1Hz,2H),1.421(s,9H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=171.95,171.73,170.16,155.95,136.52,13 6.23,136.21,131.42,130.43,127.41,127.31,123.11,121.71,121.69,11 9.84,119.82,118.81,118.80,117.91,111.14,111.13,109.72,104.93,79 .55,67.88,59.53,54.82,38.05,30.84,28.41,28.40,28.39,27.85,22.64.

[0040] Example 10 Preparation of 3S-[2-(Trp-His)]-tetrahydro-β-carboline-3-carboxylic acid (3d)

[0041] Using the procedure of Example 4, 4.32 g (91%) of the title compound was obtained from 5.6 g (8.8 mmol) of 3S-[2-(Boc-Trp-His)]-tetrahydro-β-carboline-3-carboxylic acid as a colorless solid. FT-ICR-MS: 540.2359 [M+H] + ; 1H NMR (300MHz, DMSO-d6): δ / ppm=13.071(s,1H),12.890(s,1H),11.670(s,1H),10.791(s,1H),8.731(s,1H),8.862(s,2H),8.322( s,1H),7.661(s,1H),7.581(d,J=7.6Hz,1H),7.350(d,J=7.9Hz,1H),7.331(d,J=7.9Hz,1H),7.211(d,J=7.9Hz,1H),7.201(s,1H) ,7.060(t,J=7.2Hz,1H),6.980(t,J=7.2Hz,1H),6.930(t,J=7.2Hz,1H),6.911(t,J=7.2Hz,1H),4.921(t,J=5.6Hz,1H),4.851(t ,J1=6.1Hz,1H),4.452(s,2H),3.951(t,J=5.6Hz,1H),3.313(d,J1=5.6Hz,2H),3.173(d,J1=5.6Hz,2H),3.051(t,J1=6.1Hz,2H); 13 CNMR (75MHz, DMSO-d6): δ / ppm=171.93,171.71,170.15,136.51,136.25,136.23,131.32,130.41,127.42,127.32,123.10,121.72,1 21.70,119.85,119.83,118.82,118.81,117.93,111.15,111.12,107.13,104.93,67.87,56.72,54.82,38.03,30.83,27.45,22.65.

[0042] Example 11 Evaluation of antiplatelet aggregation activity

[0043] Fresh porcine carotid artery blood was anticoagulated with 3.8% sodium citrate (1 / 9 by volume). Platelet-rich plasma (PRP) was obtained by centrifugation at 1000g for 10 minutes, and platelet-poor plasma (PPP) was obtained by centrifugation at 3000g for 10 minutes. Platelet-rich plasma was adjusted with platelet-poor plasma so that the number of platelets in platelet-rich plasma was suitable for the determination of antiplatelet aggregation activity. 3a-d was dissolved with normal saline. 0.24mL of adjusted platelet-rich plasma was added to the turbidity tube, and then 5μL of normal saline solution or normal saline solution of 3a-d (5μL, concentration of 0.1μM, 10μM, 15μM, 20μM) was added. The absorbance baseline was adjusted, and 5μL of normal saline solution of four induction factors was added to observe the maximum aggregation rate (Am) of platelets within 5 minutes. The four inducers are platelet activating factor (PAF, final concentration of 50μM), adenosine diphosphate (ADP, final concentration of 500μM), thrombin (TH, final concentration of 50IU / L) and arachidonic acid (AA, final concentration of 7.5mg / mL). The maximum aggregation rate is the value corresponding to the peak of the aggregation curve. Each concentration was measured 6 times in parallel to form a platelet aggregation curve. According to the platelet aggregation curve, the IC values ​​of 3a-d for inhibiting platelet aggregation induced by platelet activating factor, adenosine diphosphate, thrombin and arachidonic acid were determined. 50 (See Table 1). The data in Table 1 show that 3a-d inhibit ADP-induced platelet aggregation with an IC 50 The value is the smallest, and 3a-d is a selective inhibitor of ADP.

[0044] Table 1 IC of 3a-c for inhibiting platelet aggregation induced by four inducers 50 (mean ± SD, μM)

[0045] Compound ADP TH PAF AA 3a 1.24±0.05 21.55±0.27 22.55±0.28 22.58±0.26 3b 1.26±0.06 22.43±0.29 23.31±0.29 23.31±0.27 3c 1.17±0.04 22.26±0.28 21.46±0.27 21.38±0.24 3d 1.32±0.06 23.11±0.30 21.91±0.26 21.47±0.25

[0046] n=6

[0047] Example 12 Evaluation of Anti-arterial Thrombotic Activity

[0048] 1) Pull the polyethylene tube into a thin tube with an oblique end, with a fixed length of 10.0 cm, for cannulation of the right jugular vein (with a thicker diameter) and the left carotid artery (with a thinner diameter); the middle section of the polyethylene tube is fixed to a length of 8.0 cm, with the thrombus line pressed in the direction of carotid artery cannulation. The tube must be filled with heparin before cannulation.

[0049] 2) Male SD rats weighing 200±20g were acclimatized and fasted for one day before surgery. They were randomly divided into a saline group (blank control, oral dose of 0.3mL / 100g, 10 rats), an aspirin group (positive control, oral dose of 167μmol / kg, 10 rats), and a 3a-d saline solution group (oral dose of 0.1μmol / kg, 10 rats). 30 minutes after oral administration, the rats were anesthetized by intraperitoneal injection of 20% urethane solution (7mL / kg), and surgery began 2 minutes later. During the operation, the rats were placed supine on a fixed board, the neck skin was cut open, the right common carotid artery and the left jugular vein were separated, a precisely weighed silk thread was pressed down the blood vessels, the distal end was ligated, a small cut was made at the distal end of the vein, the cannula was inserted into the venous end, heparin was injected, and then the syringe for injecting heparin was removed, the thread was tied to fix it, and then the proximal end of the artery was clamped with an artery clamp, a small cut was made at the distal end of the artery, the artery end was ligated, and the artery clamp was released after the thread was fixed to establish an extracorporeal circulation bypass. After 15 minutes of circulation, the vein was cut first to observe whether the blood circulation was normal. If the blood circulation was normal, the silk thread with thrombus attached was removed from the arterial end, and the uncoagulated blood was absorbed with filter paper, and the silk thread with thrombus attached was accurately weighed. The weight of the thrombus was obtained by subtracting the weight of the silk thread from the weight of the silk thread, and the data were listed in Table 2. The thrombus weights in the table show that 3a-d effectively inhibited arterial thrombosis in rats at an oral dose of 0.1 μmol / kg (p<0.01 compared with saline and an oral dose of 16.7 μmol / kg aspirin). This was an unexpected technical effect.

[0050] Table 2 Effects of 3a-d on arterial thrombosis in rats

[0051]

[0052] a) p<0.01 compared with normal saline and oral dose of aspirin 16.7 μmol / kg; b) p>0.05 compared with normal saline; n=10.

Claims

1. 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compounds of the following structure, It is characterized in that The amino acid represented by AA in the compound is selected from L-Pro residue, L-Tyr residue, L-Gln residue or L-His, 2. A method for preparing the 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compound according to claim 1, It is characterized in that Described preparation method comprises the following steps: 1) preparing 3S-tetrahydro-β-carboline-3-carboxylic acid; 2) preparing Boc-Trp-AA, wherein AA is L-Pro residue, L-Tyr residue, L-Gln residue or L-His; 3) preparing 2-Boc-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid, wherein AA is an L-Pro residue, an L-Tyr residue, an L-Gln residue or an L-His residue; 4) preparing 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid, wherein AA is an L-Pro residue, an L-Tyr residue, an L-Gln residue or an L-His residue.

3. Use of the 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compound pair according to claim 1 in the preparation of a drug for adenosine diphosphate-induced platelet aggregation.

4. Use of the 2-Trp-AA-3S-tetrahydro-β-carboline-3-carboxylic acid compound according to claim 1 in the preparation of an anti-arterial thrombotic drug.

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