A low-molecular-weight glycosaminoglycan derivative, its pharmaceutical composition, its preparation method and application

Abstract

The invention provides a derivate of low molecular weight fucosylated glycosaminoglycan (dLFG) and a medical composition containing the dLFG or pharmaceutically acceptable salt and application of the dLFG and the medical composition for preparing medicines for treating thrombotic diseases.

Description

technical field

[0001] The invention belongs to the technical field of medicine, in particular to a low molecular weight fucosylated glycosaminoglycan derivative (derivate of Low molecular weight Fucosylated Glycosaminoglycan, dLFG) with anticoagulant activity, its preparation method, containing the dLFG or its pharmaceutical The pharmaceutical composition of the above acceptable salt, and the application of dLFG and its pharmaceutical composition in the preparation of drugs for the treatment of thrombotic diseases. Background technique

[0002] Thromboembolic diseases, including ischemic stroke, coronary heart disease, and venous thromboembolism, are the main cause of death in humans. Antithrombotic drug therapy is the basic method for clinical prevention and treatment of thrombotic diseases, but antithrombotic drugs including fibrinolytic, anticoagulant, and antiplatelet drugs all have major and common defects: bleeding tendency and serious bleeding risk.

[0003] The cla...

Examples

Embodiment 1

[0132] Preparation of low molecular weight fucosylated glycosaminoglycan derivatives (dLFG) derived from Thelenota ananas:

[0133] 1.1 Materials

[0134] Prunus ginseng (Thelenota ananas Jaeger), commercially available, eviscerated and dried body wall; benzethonium chloride, benzyl chloride, tetrabutylammonium hydroxide (TBA), N,N-dimethylformamide (DMF), hydroxide Reagents such as sodium, sodium chloride and ethanol are commercially available analytical reagents.

[0135] 1.2 Method

[0136] (1) Extraction and preparation of FGAG (FGAG from Thelenota ananas, TAG) from Thelenota ananas: Take 300 g of the dried body wall of Prunus ginseng. ) to prepare TAG, the yield is about 1.5%, the purity is 98% (HPGPC, area normalization method), and the weight average molecular weight (Mw) is 65,890Da.

[0137] (2) Preparation of TAG quaternary ammonium salt: get step (1) gained TAG1.2g and place it in a beaker, add 40mL deionized water to make it dissolve; titrate with 75mg / mL benzet...

Embodiment 2

[0168] β-elimination and depolymerization of TAG to prepare oligo-fucosylated glycosaminoglycan derivatives (dLFG) with a series of molecular weights:

[0169] 2.1 Materials:

[0170] The TAG derived from plum blossom ginseng was prepared with the method described in Example 1. Other reagents are with embodiment 1.

[0171] 2.2 Method

[0172] (1) Preparation of TAG quaternary ammonium salt: 5.02 g of TAG quaternary ammonium salt was prepared according to the method described in Example 1.

[0173] (2) Preparation of TAG benzyl esters with different degrees of benzyl esterification: Add 50 mL of DMF to the TAG quaternary ammonium salt obtained in step (1) and stir to dissolve, add 25 mL of benzyl chloride, and react under stirring at 35°C. And sample about 15mL at different reaction times, add 100mL of 1:9 (v / v) saturated NaCl-absolute ethanol to the solution respectively, centrifuge at 3500rpm for 20min, and obtain the precipitate with 50mL of 1:9 (v / v) saturated NaCl - W...

Embodiment 3

[0184] Preparation of β-elimination depolymerization products of FGAG from different sea cucumber sources:

[0185] 3.1 Materials

[0186] Apostichopus japonicus, Holothuria edulis, Ludwigothurea grisea, Holothuria leucospilota, and Holothuria nobilis were commercially available dried body walls.

[0187] 3.2 Method

[0188] (1) The dry body wall of sea cucumber imitation sea cucumber, red-bellied sea cucumber, Brazilian ginseng, yuzu sea cucumber, and black milk sea cucumber is crushed, and 300 g of the crushed matter is taken respectively, and the FGAG contained in it is extracted as described in the method (1) of Example 1, and weighed respectively. They are AJG, HEG, LGG, HLG and HNG.

[0189] (2) Take about 1 g of AJG, HEG, LGG, HLG, and HNG respectively, and prepare the β-elimination depolymerization product dLFG according to the methods (2) to (5) described in the examples, which are respectively called dAJG, dHEG, dLGG, HLG and dHNG. The basic steps of the β-elimin...

technical field

[0001] The invention belongs to the technical field of medicine, in particular to a low molecular weight fucosylated glycosaminoglycan derivative (derivate of Low molecular weight Fucosylated Glycosaminoglycan, dLFG) with anticoagulant activity, its preparation method, containing the dLFG or its pharmaceutical The pharmaceutical composition of the above acceptable salt, and the application of dLFG and its pharmaceutical composition in the preparation of drugs for the treatment of thrombotic diseases. Background technique

[0002] Thromboembolic diseases, including ischemic stroke, coronary heart disease, and venous thromboembolism, are the main cause of death in humans. Antithrombotic drug therapy is the basic method for clinical prevention and treatment of thrombotic diseases, but antithrombotic drugs including fibrinolytic, anticoagulant, and antiplatelet drugs all have major and common defects: bleeding tendency and serious bleeding risk.

[0003] The cla...

Examples

Embodiment 1

[0132] Preparation of low molecular weight fucosylated glycosaminoglycan derivatives (dLFG) derived from Thelenota ananas:

[0133] 1.1 Materials

[0134] Prunus ginseng (Thelenota ananas Jaeger), commercially available, eviscerated and dried body wall; benzethonium chloride, benzyl chloride, tetrabutylammonium hydroxide (TBA), N,N-dimethylformamide (DMF), hydroxide Reagents such as sodium, sodium chloride and ethanol are commercially available analytical reagents.

[0135] 1.2 Method

[0136] (1) Extraction and preparation of FGAG (FGAG from Thelenota ananas, TAG) from Thelenota ananas: Take 300 g of the dried body wall of Prunus ginseng. ) to prepare TAG, the yield is about 1.5%, the purity is 98% (HPGPC, area normalization method), and the weight average molecular weight (Mw) is 65,890Da.

[0137] (2) Preparation of TAG quaternary ammonium salt: get step (1) gained TAG1.2g and place it in a beaker, add 40mL deionized water to make it dissolve; titrate with 75mg / mL benzet...

Embodiment 2

[0168] β-elimination and depolymerization of TAG to prepare oligo-fucosylated glycosaminoglycan derivatives (dLFG) with a series of molecular weights:

[0169] 2.1 Materials:

[0170] The TAG derived from plum blossom ginseng was prepared with the method described in Example 1. Other reagents are with embodiment 1.

[0171] 2.2 Method

[0172] (1) Preparation of TAG quaternary ammonium salt: 5.02 g of TAG quaternary ammonium salt was prepared according to the method described in Example 1.

[0173] (2) Preparation of TAG benzyl esters with different degrees of benzyl esterification: Add 50 mL of DMF to the TAG quaternary ammonium salt obtained in step (1) and stir to dissolve, add 25 mL of benzyl chloride, and react under stirring at 35°C. And sample about 15mL at different reaction times, add 100mL of 1:9 (v / v) saturated NaCl-absolute ethanol to the solution respectively, centrifuge at 3500rpm for 20min, and obtain the precipitate with 50mL of 1:9 (v / v) saturated NaCl - W...

Embodiment 3

[0184] Preparation of β-elimination depolymerization products of FGAG from different sea cucumber sources:

[0185] 3.1 Materials

[0186] Apostichopus japonicus, Holothuria edulis, Ludwigothurea grisea, Holothuria leucospilota, and Holothuria nobilis were commercially available dried body walls.

[0187] 3.2 Method

[0188] (1) The dry body wall of sea cucumber imitation sea cucumber, red-bellied sea cucumber, Brazilian ginseng, yuzu sea cucumber, and black milk sea cucumber is crushed, and 300 g of the crushed matter is taken respectively, and the FGAG contained in it is extracted as described in the method (1) of Example 1, and weighed respectively. They are AJG, HEG, LGG, HLG and HNG.

[0189] (2) Take about 1 g of AJG, HEG, LGG, HLG, and HNG respectively, and prepare the β-elimination depolymerization product dLFG according to the methods (2) to (5) described in the examples, which are respectively called dAJG, dHEG, dLGG, HLG and dHNG. The basic steps of the β-elimin...

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