阅读说明：本技术 华法林与维生素c的分子复合物,其制备,活性和应用 (Molecular complex of warfarin and vitamin C, preparation, activity and application thereof ) 是由 彭师奇 赵明 于 2018-07-12 设计创作，主要内容包括：本发明公开了一种华法林与维生素C的摩尔比为1比2的分子复合物,公开了这种分子复合物的制备方法,公开了这种分子复合物的抗静脉血栓活性,因而本发明公开了这种分子复合物在制备抗静脉血栓药物中的应用。(The invention discloses a molecular compound with a mole ratio of warfarin to vitamin C of 1 to 2, a preparation method of the molecular compound, and anti-venous thrombosis activity of the molecular compound, and thus the invention discloses application of the molecular compound in preparation of anti-venous thrombosis medicines.)

1. A molecular complex of warfarin and vitamin C.

2. Molecular complex of warfarin and vitamin C according to claim 1, characterized in that the molar ratio of warfarin to vitamin C in the molecular complex is 1 to 2.

3. Molecular complex of warfarin and vitamin C according to claim 2, characterized in that its ESI (-) -FT-MS spectrum has a mass number of molecular ions equal to 659.06334, the mass number being equal to the molecular weight of 1 warfarin molecule and the molecular weight of 2 vitamin C molecules minus 1 proton.

4. The warfarin-vitamin C molecular complex of claim 2, characterized in that its qCID spectrum indicates that a molecular ion with a mass number of 659.06334 can be cleaved to an ion with a mass number of 307.09765, i.e., 1 warfarin minus 1 proton, and an ion with a mass number of 351.05261, i.e., 2 vitamin C minus 1 proton, under ESI (-) -FT-MS conditions, indicating that a molecular complex with a molar ratio of 1 to 2 is the only existing form of the warfarin-vitamin C molecular complex.

5. The molecular complex of warfarin and vitamin C according to claim 2, characterized in that its NOESY spectrum gives six important correlation peaks, wherein the related peak A and the related peak B are from the interaction between two enol hydroxyl protons of vitamin C and the phenolic hydroxyl protons of warfarin, the related peak C is from the interaction between the benzene ring protons of warfarin and the chiral carbon protons of vitamin C, the related peak D is from the interaction between the hydroxymethylene protons of vitamin C and the benzene ring protons of warfarin, the related peak E is from the interaction between the benzene ring protons of warfarin and the two alcohol hydroxyl protons of vitamin C, and the related peak F is from the interaction between the benzene ring protons of warfarin and the hydroxymethylene protons of vitamin C, these six peaks require that the distance between the protons be less when a warfarin molecule and two vitamin C molecules are in close proximity to each other to form a molecular complex.

6. A method of preparing a molecular complex of warfarin and vitamin C according to claim 1, comprising:

A. preparing a clear solution from 308.1mg of warfarin and 352.1mg of vitamin C in 20mL of 30-50% ethanol solution;

B. and (4) freeze-drying the clear solution to prepare freeze-dried powder.

7. The use of warfarin-vitamin C molecular complex of claim 1 for the preparation of an oral antithrombotic medicament, in particular an oral antithrombotic medicament.

8. Use of a composition of warfarin-vitamin C molecular complex of claim 1 in combination with pharmaceutically acceptable excipients for the manufacture of an oral antithrombotic medicament, in particular an oral antithrombotic medicament.

Technical Field

The invention relates to a molecular compound with a mole ratio of warfarin to vitamin C of 1 to 2, a preparation method of the molecular compound, and anti-venous thrombosis activity of the molecular compound, and thus the invention relates to application of the molecular compound in preparing anti-venous thrombosis medicines. The invention belongs to the field of biological medicine.

Background

Warfarin is the most common oral anticoagulant appearing in the prescription, and more than 150 million outpatients take warfarin in the United states alone. The main indications for warfarin are the treatment of venous thrombosis, the prevention of stroke and systemic thrombosis from atrial fibrillation, and the prevention of thrombotic complications in patients fitted with mechanical heart valves. The anticoagulant effect of warfarin comes from its inhibition of the vitamin K synthesis pathway. Besides warfarin, there is also oral anticoagulant drug approved by SDA in us 11 months 2011 that acts directly on the coagulation system. For example, dabigatran etexilate mesylate (dabigatran, pradaxa) directly inhibits thrombin, rivaroxaban (rivaroxaban, xarelto), apixaban (eliquis) and edoxaban (edoxaban, savaysa) directly inhibits factor Xa.

Systemic bleeding is not only one of the most serious complications of oral anticoagulants, but this intractable complication is characterized by a high mortality rate. Among the complications of systemic bleeding, intracranial bleeding poses the most serious threat to clinical anticoagulant therapy. This situation has attracted interest in a number of laboratory studies and large-scale phase III clinical trials. The goal of these interests is to compare the efficacy and safety of oral anticoagulants that act directly on the coagulation system with warfarin. These comparative studies concluded that although the frequency, volume and mortality rate from intracranial hemorrhage in patients treated with oral anticoagulant drugs that act directly on the coagulation system are almost lower than in patients treated with warfarin, with equivalent efficacy, the risk remains a clinical concern. Thus, elimination of systemic bleeding complications of oral anticoagulant drugs has been one of the hot spots in drug research.

The annual rate of warfarin-induced intracranial hemorrhage is between about 0.8% and 3.7%. Factors that increase the risk of warfarin-induced intracranial hemorrhage are mainly advanced age, white matter hyperintensity seen on CT and MRI scans, hypertension, history of cerebrovascular disease, black race and international normalization rates beyond the effective therapeutic range. Warfarin-induced intracranial hemorrhage has a mortality rate of 37% -49%, and most patients die within 30 days of intracranial hemorrhage. Even in survivors with concurrent intracranial hemorrhage, 61% of patients still became disabled. The risk of mortality in warfarin-treated intracranial hemorrhages was 2.2 to 2.6 times higher than in other oral anticoagulant-induced intracranial hemorrhages. Nonetheless, warfarin remains the most widely used oral anticoagulant for the prevention of thromboembolic events. A recognized difficulty faced with warfarin anticoagulation is how to control the anticoagulation effect to be both sufficient and not excessive during treatment, particularly during the initial stages of treatment. That is, optimal dosage of warfarin is critical for anticoagulant therapy. This is a clinical need and bottleneck problem for warfarin anticoagulant therapy. The core technology for solving the bottleneck problem is to greatly enlarge the anticoagulation treatment window of warfarin.

The antioxidant and anti-inflammatory effects enable various researches to associate vitamin C with the prevention of cardiovascular and cerebrovascular diseases. Several studies have confirmed that maintaining serum vitamin C levels can reduce the risk of death caused by cardiovascular disease, reduce the risk of death caused by cerebral infarction, reduce the risk of death caused by stroke, or reduce the risk of death caused by lower limb arterial disease. In the inventors' view, these studies have made vitamin C a possibility to greatly expand the anticoagulant therapeutic window of warfarin. In the course of innovatively developing this possibility, the inventors have found that making warfarin and vitamin C a molecular complex at a molar ratio of 1/2 reduces the lowest effective dose of anticoagulation of warfarin to 1/1000 at which it shows side effects. Based on these findings, the inventors have proposed the present invention.

Disclosure of Invention

The first aspect of the present invention provides a molecular complex of warfarin and vitamin C in a molar ratio of 1/2. The molecular ion mass number of its ESI (-) -FT-MS spectrum is 659.06334, which is equal to the molecular weight of 1 warfarin molecule and the molecular weight of 2 vitamin C molecules minus 1 proton.

The qCID spectrum of the molecular complex of warfarin and vitamin C shows that under ESI (-) -FT-MS, a molecular ion with the mass number of 659.06334 can be cleaved into an ion with the mass number of 307.09765, namely the mass number of 1 warfarin minus 1 proton, and an ion with the mass number of 351.05261, namely the mass number of 2 vitamin C minus 1 proton, which indicates that the molecular complex with the molar ratio of 1 to 2 is the only existing form of the molecular complex of warfarin and vitamin C.

The NOESY spectrum of the molecular compound of warfarin and vitamin C of the invention gives six important related peaks, wherein the related peak A and the related peak B are from the interaction between two enol hydroxyl protons of the vitamin C and phenolic hydroxyl protons of warfarin, the related peak C is from the interaction between benzene ring protons of warfarin and chiral carbon protons of the vitamin C, and the related peak D is from hydroxymethylene protons of the vitamin C and warfarinThe six related peaks require that the distance between protons is less than that when one warfarin molecule and two vitamin C molecules approach each other to form a molecular complex, the proton of the benzene ring of the warfarin is less than that when the two vitamin C molecules are close to each other to form a molecular complex

The second aspect of the present invention provides a process for preparing a molecular complex of warfarin and vitamin C in a molar ratio of 1/2, comprising the following two steps:

(1) 308.1mg of warfarin, 352.1mg of vitamin C and 10mL-30mL, preferably 20mL of ethanol aqueous solution (concentration: concentration

30% -60%, preferably 50%) to prepare a clear solution;

(2) and (4) freeze-drying the clear solution to prepare freeze-dried powder.

A third aspect of the invention is to provide an ESI (-) -FT-MS spectrum of molecular complexes of warfarin and vitamin C at a molar ratio of 1/2.

A fourth aspect of the present invention is to provide a NOESY profile of a molecular complex of warfarin and vitamin C in a molar ratio of 1/2.

A fifth aspect of the present invention is to describe the antithrombotic effect of a molecular complex of warfarin and vitamin C in a molar ratio of 1/2.

Drawings

FIG. 1 ESI (-) -FT-MS and qCID spectra of molecular complexes of warfarin and vitamin C at a molar ratio of 1/2,

FIG. 2. NOESY spectrum of molecular complex of warfarin and vitamin C at a molar ratio of 1/2,

FIG. 3. Effect of 1 to 2 molar ratio of warfarin to vitamin C molecular complex on rat PT,

FIG. 4 shows the bleeding status of rats treated with molecular complex of warfarin and vitamin C at a molar ratio of 1 to 2.

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.