CN111793067A

CN111793067A - Synthesis, activity and application of dihydroxymethyl tetrahydrocarboline-3-formyl-The

Info

Publication number: CN111793067A
Application number: CN201910273805.6A
Authority: CN
Inventors: 赵明; 彭师奇; 张筱宜; 郤思远
Original assignee: Capital Medical University
Current assignee: Capital Medical University
Priority date: 2019-04-07
Filing date: 2019-04-07
Publication date: 2020-10-20
Anticipated expiration: 2039-04-07
Also published as: CN111793067B

Abstract

The invention discloses (3S) -1, 1-dihydroxymethyl-tetrahydro-beta-carboline-3-formyl-The, a preparation method of The compound and The anti-arterial thrombosis activity of The compound. Therefore, the invention discloses the application of the compound in the preparation of the antithrombotic drug.

Description

Synthesis, activity and application of dihydroxymethyl tetrahydrocarboline-3-formyl-The

Technical Field

The invention relates to (3S) -1, 1-dihydroxymethyl-tetrahydro-beta-carboline-3-formyl-The with The following structure, a preparation method thereof and anti-arterial thrombosis activity thereof. The invention thus relates to the use of the compounds in the preparation of antithrombotic medicaments. The invention belongs to the field of biological medicine.

Background

The mortality and morbidity of thrombi, both in developed and developing countries, is rising year by year. Arterial thrombosis has become the first fatal thrombotic disease in China. It is well known that almost all malignant tumors can metastasize through thrombi. A large number of clinical studies have shown that more than 80% of cases of malignant tumors have complicated arterial thrombosis. The invention relates to a medicament with anti-arterial thrombosis activity, which is one of the leading fields in the field of biological medicines. Also, the team of the inventor who has been present over the past decade has paid a hard creation for inventing drugs with anti-arterial thrombosis. Meanwhile, the inventor finds that (3S) -1, 1-dihydroxymethyl-tetrahydro-beta-carboline-3-carboxylic acid is not only orally effective, but also has a dosage as low as 10 nmol/kg. Based on this finding, the inventors have proposed the present invention.

Disclosure of Invention

The first aspect of The present invention is to provide (3S) -1, 1-dimethylol-tetrahydro- β -carboline-3-formyl-The of The following structure.

The second aspect of The present invention provides a process for preparing (3S) -1, 1-dimethylol-tetrahydro- β -carboline-3-formyl-The, which comprises 7 steps:

the first step is to prepare (3S) -1, 1-dihydroxymethyl-tetrahydro-beta-carboline-3-carboxylic acid;

the second step is to prepare (3S) -1, 1-dihydroxymethyl-tetrahydro-beta-carboline-3-carboxylic acid methyl ester;

the third step is to prepare (3S) -1, 1-di (tert-butyldimethylsilyloxy) methyl-tetrahydro-beta-carboline-3-carboxylic acid methyl ester;

the fourth step is to prepare (3S) -1, 1-di (tert-butyldimethylsilyloxy) methyl-tetrahydro-beta-carboline-3-carboxylic acid;

the fifth step is to prepare (3S) -1, 1-di (tert-butyldimethylsilyloxy) methyl-tetrahydro-beta-carboline-3-formyl-The-OMe by adopting Dicyclohexylcarbodiimide (DCC) as a condensing agent and 1-hydroxybenzotriazole (HOBt) as a catalyst through liquid phase condensation;

the sixth step is to prepare (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro-beta-carboline-3-formyl-The;

the seventh step is to prepare (3S) -1, 1-dihydroxymethyl-beta-carboline-3-formyl-The.

The third aspect of The present invention is to evaluate The anti-arterial thrombotic activity of (3S) -1, 1-dihydroxymethyl-tetrahydro- β -carboline-3-formyl-The.

Drawings

FIG. 1 is a synthetic route for (3S) -1, 1-dihydroxymethyl-tetrahydro- β -carboline-3-formyl-The: i) h₂SO₄(ii), (1, 3-dihydroxyacetone); ii) thionyl chloride, CH₃OH; iii) tert-butyldimethylchlorosilane (TBDMSCl), imidazole (imidazole); iv) CH₃OH, NaOH; v) DCC, HOBt, N-methylmorpholine (NMM); vi) hydrogen chloride in ethyl acetate.

Detailed Description

To further illustrate the invention, a series of examples are given below. These examples are purely illustrative and are intended to be a detailed description of the invention only and should not be taken as limiting the invention.

EXAMPLE 1 preparation of (3S) -1, 1-dimethylol-tetrahydro-beta-carboline-3-carboxylic acid (1)

To 50mL of distilled water was added 3.06g (15mmol) of L-tryptophan, and the mixture was stirred well to suspend the tryptophan. And slowly dropwise adding concentrated sulfuric acid into the suspension under an ice bath condition until the L-tryptophan is completely dissolved. Then, 1.62g (18mmol) of 1, 3-dihydroxyacetone was added to the solution and reacted at room temperature for 84 hours. TLC showed the disappearance of L-tryptophan (ethyl acetate/water/glacial acetic acid, 4/1/1). Filtration and washing of the filter cake with ice water gave 2.90g (70%) of the title compound as a yellow powder.

EXAMPLE 2 preparation of (3S) -1, 1-dimethylol-tetrahydro-beta-carboline-3-carboxylic acid methyl ester (2)

4.7mL of thionyl chloride was slowly added dropwise to 50mL of methanol in ice bath, and after stirring for 30 minutes, 5.0g (18mmol) of (3S) -1, 1-dimethylol-tetrahydro- β -carboline-3-carboxylic acid (1) was added thereto. Stir until compound 1 is completely dissolved. Thereafter, the mixture was stirred at room temperature for 10 hours. TLC showed the disappearance of compound 1 (dichloromethane/methanol, 20: 1). The reaction mixture was concentrated under reduced pressure, and the residue was triturated with ethyl acetate. The supernatant was removed and the residue was triturated with ethyl acetate. The resulting brown-yellow solid was dissolved in 100mL of ethyl acetate. The solution was sequentially treated with saturated NaHCO₃Aqueous solution (30 mL. times.3), saturatedAnd an aqueous solution of NaCl (30 mL. times.3), and the ethyl acetate layer was dried over anhydrous sodium sulfate for 12 hours. Filtration and concentration of the filtrate under reduced pressure gave 4.73g (90%) of the title compound as a yellow solid. ESI-MS (M/e):291[ M + H]⁺。

Example 3 preparation of (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro-beta-carboline-3-carboxylic acid methyl ester (3)

To 50mL of anhydrous N, N-Dimethylformamide (DMF) was added 4.35g (15mmol) of methyl 1, 1-dimethylol- β -carboline-3-carboxylate (2). Stirring until the compound 2 is completely dissolved. Thereafter, 3.66g (64.8mmol) of imidazole was added to the solution under ice-bath. Stirring until imidazole is completely dissolved. Thereafter, 6.79g (45mmol) of t-butyldimethylsilyl chloride (TBDMSCl) was added to the solution. The reaction mixture was stirred at room temperature for 6 hours. TLC showed the disappearance of compound 2 (petroleum ether/ethyl acetate, 20/1). The reaction mixture was concentrated under reduced pressure, and the residue was diluted with 100mL of saturated aqueous NaCl solution. The resulting solution was extracted with ethyl acetate (60 mL. times.3), and the ethyl acetate layer was then successively replaced with saturated NaHCO₃The mixture was washed with an aqueous solution (30 mL. times.3) and a saturated aqueous NaCl solution (30 mL. times.3), and dried over anhydrous sodium sulfate for 12 hours. Filtering, and concentrating the filtrate under reduced pressure. The yellow residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate, 10/1) to give 4.82g (88%) of the title compound as a colorless solid. ESI-MS (M/e):519[ M + H]⁺。

Example 4 preparation of (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro-beta-carboline-3-carboxylic acid (4)

To 40mL of a tetrahydrofuran/methanol (v/v, 1/1) mixed solution was added 5.18g (10mmol) of methyl (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro- β -carboline-3-carboxylate (3). Stirring until compound 3 is completely dissolved. An aqueous NaOH solution (4M) was added dropwise to the solution in an ice bath to adjust the pH of the reaction solution to 13. After stirring for 30 min, TLC showed the disappearance of compound 3 (petroleum ether/ethyl acetate, 20/1). Adding saturated KHSO dropwise into the reaction mixture under ice bath₄The pH of the aqueous solution was adjusted to 7. The reaction mixture was concentrated under reduced pressure. Under ice-bath, the residue was added dropwise with saturated KHSO₄The pH of the aqueous solution was adjusted to 2. The resulting solution was extracted with ethyl acetate (30 mL. times.3), and the ethyl acetate layer was washed with saturated aqueous NaCl solution and anhydrous sodium sulfateDrying for 12 hours. Filtration and concentration of the filtrate under reduced pressure gave 4.64g (90%) of the title compound as a colorless powder. ESI-MS (M/e) 505[ M + H ]]⁺。

EXAMPLE 5 preparation of (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro- β -carboline-3-formyl-The-OMe (5)

To 50mL of DMF were added 5.04g (10mmol) of (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro- β -carboline-3-carboxylic acid (4), 2.47g (12mmol) of N, N' -Dicyclohexylcarbodiimide (DCC) and 1.62g (12mmol) of 1-hydroxybenzotriazole (HOBt). Stir for 30 minutes in an ice bath. Then, 2.48g (11mmol) of HCl. The-OMe was added to The reaction mixture. Subsequently, N-methylmorpholine (NMM) was added dropwise to the reaction mixture to adjust the pH of the reaction mixture to 9. After stirring at room temperature for 6 h, TLC showed the disappearance of compound 4 (dichloromethane/methanol, 30/1). Dicyclohexylurea (DCU) was filtered off and the filtrate was concentrated under reduced pressure. The residue was dissolved in 60mL of ethyl acetate and the solution was filtered to remove DCU. The filtrate was sequentially treated with 5% NaHCO₃Aqueous solution (30 mL. times.3), saturated aqueous NaCl solution (30 mL. times.3), 5% KHSO₄Aqueous wash (30 mL. times.3), saturated aqueous NaCl wash (30 mL. times.3), 5% NaHCO₃The mixture was washed with an aqueous solution (30 mL. times.3) and with a saturated aqueous NaCl solution (30 mL. times.3), and dried over anhydrous sodium sulfate for 12 hours. Filtration and concentration of the filtrate under reduced pressure gave a yellow powder which was purified by silica gel column chromatography (petroleum ether/ethyl acetate, 10/1) to give 5.4g (80%) of the title compound as a colourless powder. ESI-MS (M/e):675[ M + H]⁺。

EXAMPLE 6 preparation of (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro- β -carboline-3-formyl-The (6)

Using The method of example 4, 2.64g (80%) of The title compound were obtained as colorless powder from 3.72g (5mmol) of (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro- β -carboline-3-formyl-The-OMe (5). ESI-MS (M/e):661[ M + H]⁺。

EXAMPLE 7 preparation of (3S) -1, 1-dihydroxymethyl-beta-carboline-3-formyl-The (7)

0.09g (0.1mmol) of (3S) -1, 1-bis (tert-butyldimethylsilyloxy) methyl-tetrahydro- β -carboline-3-formyl-The (6) was dissolved in 20mL of ethyl acetate (4M) of hydrogen chloride and reacted for 4 hours under ice bath. TLC showed chemical combinationThe product (6) disappeared (ethyl acetate: water: glacial acetic acid 4:1:1), the reaction mixture was concentrated under reduced pressure, the residue was dissolved in anhydrous ethyl acetate, and the resulting solution was concentrated under reduced pressure. This operation was repeated 3 times. The obtained white powdery substance was sufficiently washed with dehydrated ether and then subjected to¹⁸C column chromatography gave 0.045g (60%) of the title compound as a yellow solid, ESI-MS (M/e):431[ M-H]-，¹H-NMR(300MHz,DMSO-d₆):/ppm＝10.58(s,1H),8.26(d,J＝7.2Hz,1H),7.86(td,J₁＝7.8Hz,J₂＝1.6Hz,2H),7.00(dt,J₁＝7.8Hz,J₂＝1.6Hz,2H),6.96(m,2H),4.70(s,2H),4.21(m,1H),3.87(m,1H),3.73(m,1H),3.53(m,3H),3.06(m,3H),2.90(m,1H),2.17(t,J＝7.2Hz,2H),2.02(m,1H),1.90(m,1H),0.99(t,J＝7.2Hz,3H)。

EXAMPLE 8 evaluation of in vivo anti-arterial Thrombus Activity inhibition by Compound 7

Experimental animals: SD rats, male, 200 + -20 g, purchased from Experimental animals technologies, Inc. of Wei Tongli, Beijing.

Administration dose and administration mode: the oral dose of Compound 7 of the present invention was 0.01. mu. mol/kg, the oral dose of a physiological saline solution of positive control aspirin was 167. mu. mol/kg, and the negative control was physiological saline.

The experimental method comprises the following steps: the experiment adopts a rat common carotid artery-external jugular vein in vitro bypass circulation silk thread model.

And (3) experimental operation: the bypass circulation is composed of three sections of polyethylene rubber tubes, wherein the length of the middle section is 80.0mm, the inner diameter is 3.5mm, the lengths of the tubes at the two ends are 100.0mm, the inner diameter is 1.0mm, and the outer diameter is 2.0 mm. To allow insertion into the rat carotid artery or vein, one end of the tube was pulled into a sharp tube. And selecting silk threads with weight of 4.0 +/-0.1 mg, length of 60.0mm and uniform surface roughness as thrombus carriers. Silanizing the three sections of polyethylene rubber tubes with 1% of silicon ether solution, placing the silk threads at one end of the middle section of polyethylene tube after completely airing, assembling the three sections of polyethylene tubes, and sealing and fixing the joints of the three sections of polyethylene tubes by using a sealing film.

After 30 minutes of oral administration of the drug to rats, 20% urethane solution (0.7mL/100g) was intraperitoneally injected for anesthesia. The anesthetized rat was supine fixed on the rat plate, the neck skin was cut open, the right common carotid artery and the left external jugular vein were separated, and the distal ends of the right common carotid artery and the left external jugular vein were ligated with an operation wire. A bevel opening is cut on the left external jugular vein, a sharp tube of a bypass pipeline filled with the heparin sodium solution is inserted into the opening of the left external jugular vein through the bevel opening, a polyethylene tube and a vein are fixed by an operation line, and a fixed amount (0.1mL/100g) of heparin sodium is added into the other end of the polyethylene tube for anticoagulation by a syringe. Clamping the proximal end of the right carotid artery by an artery clamp, cutting an oblique opening in the right carotid artery, taking down a section of the syringe of the bypass tube, inserting the syringe into the proximal end of the right carotid artery, fixing the polyethylene tube with the artery blood vessel by an operation line, loosening the artery clamp, and enabling blood flow to flow from the right artery to the polyethylene tube and flow into the left vein to establish extracorporeal circulation. The blood flow is maintained smooth, the silk thread with thrombus is taken out after the systemic circulation is carried out for 15 minutes, the blood on the silk thread is sucked by filter paper, and then the wet weight of the thrombus is weighed and recorded.

The experimental results are as follows: the results are shown in Table 1.

TABLE 1 anti-arterial thrombotic Activity of Compound 7

n-10, a) to saline ratio P <0.01 to aspirin ratio P >0.05.

The results show that the anti-arterial thrombotic activity of compound 7 at 0.01 μmol/kg oral dose is not only significantly stronger than normal saline but also not significantly different from aspirin at 167 μmol/kg oral dose. It can be seen that compound 7 of the present invention has outstanding technical effects.

Claims

1.(3S) -1, 1-dihydroxymethyl-beta-carboline-3-formyl-The with The following structure,

2. a process for The preparation of (3S) -1, 1-dimethylol- β -carboline-3-formyl-The according to claim 1, which comprises 7 steps:

3. Use of (3S) -1, 1-dimethylol-tetrahydro- β -carboline-3-formyl-The according to claim 1 for The preparation of an anti-arterial thrombosis medicament.