WO2023174090A1 - Cocrystal of dabigatran etexilate and method for preparing same - Google Patents

Abstract

The present invention relates to a cocrystal of dabigatran etexilate and a method for preparing same, also relates to a pharmaceutical composition comprising the cocrystal, and further relates to use of the cocrystal in preparing an anticoagulant drug. The cocrystal comprises dabigatran etexilate and a cocrystal former, the cocrystal former being a specific type of organic acid. The cocrystal has the advantages of high solubility, high in vitro dissolution rate, high in vivo relative bioavailability, and the like.

Description

Dabigatran etexilate cocrystal and preparation method thereof Technical field

The invention belongs to the field of medical technology and relates to a co-crystal of dabigatran etexilate and a preparation method, a pharmaceutical composition containing the co-crystal, and the application of the co-crystal in the preparation of anticoagulant drugs.

Background technique

Dabigatran etexilate (chemical name: 3-[[[2-[[[4-[[(hexyloxy)carbonyl]amino]iminomethyl]phenyl]amino]methyl]-1 -Methyl-1H-benzimidazol-5-yl]carbonyl](pyridin-2-yl)amino]propionate ethyl ester, formula 1) is a new thrombin inhibitor used to prevent stroke and thrombosis. Dabigatran etexilate is the prodrug of dabigatran. Its mechanism of action is that after oral administration, it is rapidly converted into biologically active dabigatran by non-specific esterases in the intestine, liver and blood. The latter competitively binds to the activation site of thrombin to inhibit thrombin, which significantly reduces the risk of bleeding while ensuring anticoagulant efficacy. Compared with the traditional anticoagulant warfarin, dabigatran etexilate is more effective and safer, does not require special medication monitoring, and has good application prospects. However, the water solubility of dabigatran etexilate is extremely poor, less than 3 μg/ml, and has significant pH dependence. It is easy to crystallize and precipitate in the gastrointestinal tract. Due to its insoluble and weakly alkaline characteristics, it causes bioavailability problems. greatly limits its clinical application.

Pharmaceutical cocrystals are pharmaceutical active ingredients (API) and cocrystal ligands (cocrystals former, CCF) in a fixed stoichiometric ratio through hydrogen bonds, van der Waals forces, π-π conjugation and halogen bonds and other non-conjugated substances. A multi-component system composed of covalent bonds. Drug co-crystals can improve the physical and chemical properties of drugs, such as melting point, stability, solubility and bioavailability, without changing the chemical structure of the drug. In recent years, drug cocrystals have become a new strategy for drug research and development, showing attractive application prospects in the field of medical biology.

Contents of the invention

The inventor unexpectedly obtained a co-crystal of dabigatran etexilate, which improved the solubility, dissolution and in vivo relative bioavailability of poorly soluble dabigatran etexilate compared with the prior art.

The present invention provides a co-crystal comprising dabigatran etexilate and a co-crystal ligand, where the co-crystal ligand is a pharmaceutically acceptable organic acid.

The present invention also provides a pharmaceutical composition, which contains the co-crystal and a pharmaceutically acceptable carrier or excipient.

The invention also provides a method for preparing the eutectic, including:

a) Dissolve dabigatran etexilate and co-crystal ligand in the solvent;

b) Cool down until crystals precipitate;

c) Optionally, the crystals are isolated and optionally dried.

The present invention also provides another method for preparing the eutectic, including:

a) Dissolve dabigatran etexilate and co-crystal ligand in the solvent and stir;

b) Slowly add water dropwise to the above solution and stir;

c) Slowly evaporates;

d) Optionally, dry the solid sample.

The present invention also provides the use of the cocrystal in preparing drugs for anticoagulation.

The present invention also provides the use of the cocrystal in the preparation of drugs for treating and/or preventing thrombosis.

Detailed description of the invention

In one aspect, the present application provides a co-crystal comprising dabigatran etexilate and a co-crystal ligand, the co-crystal ligand being an organic acid selected from p-hydroxybenzoic acid and 2,4-dihydroxybenzoic acid. , succinic acid, benzoic acid, tartaric acid, maleic acid, fumaric acid or any combination thereof.

In certain embodiments, the organic acid is p-hydroxybenzoic acid or 2,4-dihydroxybenzoic acid.

In certain embodiments, the organic acid is 2,4-dihydroxybenzoic acid.

In certain embodiments, in the co-crystal, the molar ratio of dabigatran etexilate to the co-crystal ligand is about 1:0.5-50, preferably about 1:0.5-25, more preferably about 1:0.5 ~10, more preferably about 1:0.5~5.

In certain embodiments, the co-crystal ligand is p-hydroxybenzoic acid, 2,4-dihydroxybenzoic acid, or succinic acid. In certain embodiments, the molar ratio of dabigatran etexilate to the co-crystal ligand is about 1:0.5～5, about 1:0.8～4, about 1:0.8～3, about 1:0.8～2.5 , about 1:0.8～1.5, about 1:0.8～1.2, for example, about 1:1.

In certain embodiments, the co-crystal is a co-crystal of dabigatran etexilate and p-hydroxybenzoic acid or 2,4-dihydroxybenzoic acid, and the molar ratio of dabigatran etexilate to the co-crystal ligand is About 1:1.

In certain embodiments, in the co-crystal, the co-crystal ligand is 2,4-dihydroxybenzoic acid, and the The molar ratio of dabigatran etexilate to cocrystal ligand is approximately 1:1.

In certain embodiments, the co-crystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid has an X-ray powder diffraction pattern expressed at an angle of 2θ obtained using Cu-Kα radiation at 20.7 There are characteristic peaks at °±0.2° and 24.9°±0.2°.

In certain embodiments, the co-crystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid, in an X-ray powder diffraction pattern expressed at a 2θ angle obtained using Cu-Kα radiation, also has There are characteristic peaks at 17.9°±0.2°, 22.1°±0.2°, 23.2°±0.2°, and 24.1°±0.2°.

In certain embodiments, the co-crystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid, in an X-ray powder diffraction pattern expressed at a 2θ angle obtained using Cu-Kα radiation, also has There are characteristic peaks at 21.9°±0.2°, 26.4°±0.2°, 27.1°±0.2°, and 28.5°±0.2°.

In certain embodiments, the co-crystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid, in an X-ray powder diffraction pattern expressed at a 2θ angle obtained using Cu-Kα radiation, also has There are characteristic peaks at 9.5°±0.2°, 11.0°±0.2°, 22.6°±0.2°, and 27.1°±0.2°.

In certain embodiments, the co-crystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid, in an X-ray powder diffraction pattern expressed at a 2θ angle obtained using Cu-Kα radiation, also has 17.8°±0.2°, 18.7°±0.2°, 25.7°±0.2°, 9.9°±0.2°, 19.1°±0.2°, 18.9°±0.2°, 20.3°±0.2°, 21.5°±0.2°, 17.0° There are characteristic peaks at ±0.2° and 28.3°±0.2°.

In certain embodiments, the cocrystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid has substantially the same X-ray powder diffraction pattern as shown in Figure 5 using Cu-Kα radiation.

In certain embodiments, as compared to the dabigatran etexilate drug substance, the dabigatran etexilate has a higher concentration of 2,4 -The cocrystal of dihydroxybenzoic acid has no characteristic peaks at 8.9°±0.2° and 15.5°±0.2°, and has characteristic peaks at 20.7°±0.2° and 24.9°±0.2°.

In certain embodiments, the dabigatran etexilate drug substance has an X-ray powder diffraction pattern substantially the same as shown in Figure 6 using Cu-Kα radiation.

In certain embodiments, the co-crystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid exhibits melting in the range of 120°C to 130°C as measured by differential scanning calorimetry (DSC) Endothermic phenomenon.

In certain embodiments, the co-crystal of dabigatran etexilate and 2,4-dihydroxybenzoic acid has substantially the same differential scanning volume as shown in Figure 8 as measured by differential scanning calorimetry. Thermal curve.

In one aspect, the present application also provides a pharmaceutical composition comprising the co-crystal described in any one of the above, and a pharmaceutically acceptable carrier or excipient.

In certain embodiments, the co-crystal is present in the pharmaceutical composition in an anticoagulant effective amount.

In one aspect, the application also provides a method for preparing the cocrystal of the present invention, comprising:

a) Dissolve dabigatran etexilate and co-crystal ligand in the solvent;

b) Cool down until crystals precipitate;

c) Optionally, isolate the crystals and optionally dry the crystals.

In certain embodiments, the solvent is selected from water, alcohol solvents, ester solvents, ketone solvents, ether solvents, nitrile solvents, alkane solvents, halogenated alkane solvents, or any combination thereof.

In certain embodiments, the alcoholic solvent is selected from methanol, ethanol, isopropyl alcohol, or any combination thereof.

In certain embodiments, the ester solvent is selected from methyl acetate, ethyl acetate, propyl acetate, or any combination thereof.

In certain embodiments, the ketone solvent is selected from acetone, methyl isobutyl ketone, or combinations thereof.

In certain embodiments, the ether solvent is selected from methyl tert-butyl ether, cyclopentyl methyl ether, tetrahydrofuran, or any combination thereof.

In certain embodiments, the nitrile solvent is acetonitrile.

In certain embodiments, the alkane solvent is n-hexane.

In certain embodiments, the haloalkane solvent is methylene chloride.

In certain embodiments, the solvent is selected from methanol, ethanol, tert-butyl methyl ether, n-hexane, water, or any combination thereof.

In one aspect, the application also provides another method for preparing the cocrystal of the present invention, including:

a) Dissolve dabigatran etexilate and co-crystal ligand in a non-aqueous solvent and stir;

b) Slowly add water dropwise to the above solution and stir;

c) Slowly volatilize to obtain a solid;

d) Optionally, dry the solid.

In certain embodiments, the solvent is selected from alcohol solvents, ester solvents, ketone solvents, ether solvents, nitrile solvents, alkane solvents, halogenated alkane solvents, or any combination thereof.

In certain embodiments, the alcoholic solvent is selected from methanol, ethanol, isopropyl alcohol, or any combination thereof.

In certain embodiments, the ester solvent is selected from methyl acetate, ethyl acetate, propyl acetate, or any combination thereof.

In certain embodiments, the ketone solvent is selected from acetone, methyl isobutyl ketone, or combinations thereof.

In certain embodiments, the ether solvent is selected from methyl tert-butyl ether, cyclopentyl methyl ether, tetrahydrofuran, or any combination thereof.

In certain embodiments, the nitrile solvent is acetonitrile.

In certain embodiments, the alkane solvent is n-hexane.

In certain embodiments, the haloalkane solvent is methylene chloride.

In certain embodiments, the solvent is n-hexane.

This application also provides the use of the cocrystal of the present invention in the preparation of anticoagulant drugs.

This application also provides the use of the cocrystal of the present invention in the preparation of medicaments for the treatment and/or prevention of thrombosis.

The application also provides the use of the cocrystal of the invention in the preparation of medicaments for the treatment and/or prevention of venous thromboembolism (VTE), deep vein thrombosis (DVT), pulmonary embolism (PE), stroke and systemic embolism.

Definition of Terms

In the present invention, unless otherwise stated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Meanwhile, in order to better understand the present invention, definitions and explanations of relevant terms are provided below.

As used herein, the term "substantially the same" used to qualify a figure is intended to mean that a person skilled in the art would consider the figure to be identical to the reference figure, taking into account acceptable deviations in the art. Such deviations may be caused by factors known in the art related to instrumentation, operating conditions, and human factors. For example, those skilled in the art will appreciate that endothermic onset and peak temperatures measured by differential scanning calorimetry (DSC) can vary significantly from experiment to experiment. In some embodiments, two graphs are considered to be substantially the same when the positions of characteristic peaks of the two graphs do not vary by more than ±5%, ±4%, ±3%, ±2%, or ±1%. For example, one skilled in the art can easily identify whether two X-ray diffraction patterns or two DSC patterns are substantially the same. In some embodiments, two X-ray diffraction patterns are considered to be substantially the same when the 2θ angle of the characteristic peaks of the two X-ray diffraction patterns does not change by more than ±0.3°, ±0.2°, or ±0.1°.

As used herein, the term "effective amount" refers to an amount sufficient to achieve the desired therapeutic or preventive effect, for example, an amount that reduces symptoms associated with the disease to be treated (e.g., thrombosis), or is effective to avoid, reduce, An amount that prevents or delays the onset of a disease, such as a blood clot. Determining such effective amounts is within the ability of those skilled in the art. Generally speaking, the daily dosage of the cocrystal of the present invention for treatment can be about 1 to 1000 mg.

As used herein, the term "treatment" is intended to alleviate, lessen, ameliorate, or eliminate the disease state or disorder targeted. If a subject receives a therapeutic amount of the co-crystal or pharmaceutical composition in accordance with the methods described herein, the subject exhibits an observable and/or detectable reduction in one or more signs and symptoms. or improved, the subject was successfully "treated". It should also be understood that treatment of a disease state or disorder includes not only complete treatment; It does not achieve a complete cure, but achieves some biologically or medically relevant results.

As used herein, the term "prevention" is intended to avoid, reduce, prevent or delay the occurrence of a disease or disease-related symptoms before such disease or disease-related symptoms occur prior to the administration of the relevant drug. "Prevention" does not necessarily require completely preventing the occurrence of a disease or disease-related symptoms. For example, administration of a drug can reduce the risk of a subject developing a specific disease or disease-related symptoms, or attenuate the severity of subsequent related symptoms. All can be considered as "preventing" the emergence or development of the disease.

As used herein, the term "about" shall be understood to mean within the normal tolerance range in the art, for example, about may be understood to mean ±10%, ±9%, ±8%, ±7%, ± Within 6%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, ±0.1%, ±0.05% or ±0.01%. All numerical values provided herein are modified by the term "about" unless otherwise apparent from context.

As used herein, the term "pharmaceutically acceptable carrier or excipient" refers to a diluent, addendum, or vehicle that is administered with a therapeutic agent and that is, within the scope of sound medical judgment, suitable Exposure to human and/or other animal tissue without undue toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit/risk ratio.

Pharmaceutically acceptable carriers that may be used in the pharmaceutical compositions of the present invention include, but are not limited to, sterile liquids, such as water, and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is an exemplary carrier when the pharmaceutical composition is administered intravenously. Physiological saline and aqueous glucose and glycerol solutions may also be used as liquid carriers, particularly for injections. Suitable excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skimmed milk powder, glycerin, propylene glycol, water, ethanol wait. The composition may also contain a small amount of a wetting agent, an emulsifier, a pH buffer, a preservative, an antioxidant, a flavoring agent, a fragrance, a co-solvent, a solubilizer, an osmotic pressure regulator, a coloring agent, etc., if necessary. Oral formulations may contain standard carriers such as binders, fillers, disintegrating agents, lubricants, and the like.

The pharmaceutical composition of the present invention can be administered by methods known in the art, such as but not limited to administration in any of the following ways: oral administration, spray inhalation, rectal administration, nasal administration, buccal administration, topical administration, parenteral administration The drug is administered by subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or by means of an explanted reservoir. Among them, oral, intramuscular or intravenous injection administration is preferred.

For these routes of administration, the pharmaceutical compositions of the present invention can be administered in suitable dosage forms.

The dosage form may be a solid preparation, a semi-solid preparation, a liquid preparation or a gaseous preparation, including but not limited to tablets, Capsules, powders, granules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions , suspensions, elixirs, syrups.

The pharmaceutical composition of the present invention can be prepared by any method well known in the art, such as by mixing, dissolving, granulating, sugar coating, grinding, emulsifying, lyophilizing and other processes.

beneficial effects

Compared with the dabigatran etexilate API, the co-crystal of dabigatran etexilate provided by the present invention has one or more of the following advantages:

1) High solubility;

2) High in vitro dissolution;

3) High relative bioavailability in the body.

In addition, the present invention utilizes the principles of crystal engineering, combined with the characteristics of the presence of amide groups in the structure of dabigatran etexilate, and uses a cooling crystallization method or a solvent method to increase the opportunity for intermolecular contact and accelerate the eutectic formation rate.

The method for preparing the eutectic provided by the invention has one or more of the following advantages:

1) Simple operation;

2) Short preparation time;

3) The quality of the obtained eutectic is stable;

4) The method is highly controllable and reproducible;

5) Low cost.

Description of the drawings

Figure 1 shows the equilibrium solubility results of dabigatran etexilate and the product obtained in Example 1 of the present invention;

Figure 2 shows the equilibrium solubility results of dabigatran etexilate and the product obtained in Example 2 of the present invention;

Figure 3 shows the in vitro dissolution curves of dabigatran etexilate and the product obtained in Example 1 of the present invention;

Figure 4 shows the in vitro dissolution curves of dabigatran etexilate and the product obtained in Example 2 of the present invention;

Figure 5 shows the XRPD pattern of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1 of the present invention;

Figure 6 shows the XRPD pattern of dabigatran etexilate API;

Figure 7 shows the dabigatran etexilate/2,4-dihydroxybenzoic acid co-crystal prepared in Example 1 of the present invention and dabigatran etexilate, 2,4-dihydroxybenzoic acid, dabigatran etexilate/ XRPD comparison pattern of physical mixture of 2,4-dihydroxybenzoic acid;

Figure 8 shows the DSC chart of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1 of the present invention. Spectrum;

Figure 9 shows the DSC comparison spectra of dabigatran etexilate, 2,4-dihydroxybenzoic acid, and the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1 of the present invention;

Figure 10 shows the FTIR spectrum of dabigatran etexilate;

Figure 11 shows the FTIR spectrum of 2,4-dihydroxybenzoic acid;

Figure 12 shows the FTIR spectrum of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1 of the present invention.

Detailed ways

The embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will understand that the following examples and experimental examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. If the specific conditions are not indicated in the examples and test examples, they shall be carried out according to the conventional conditions or the conditions recommended by the manufacturer. If the reagents or instruments used are not indicated by the manufacturer, they are all conventional products that can be purchased commercially.

Dabigatran etexilate used in the embodiments of the present invention is either commercially available (Hanxiang Biotechnology Co., Ltd., product number BCP02864, batch number 20200705), or can be prepared by referring to existing technologies. All co-crystal ligands used are commercially available. For example, p-hydroxybenzoic acid, 2,4-dihydroxybenzoic acid, succinic acid, nicotinamide, benzamide, and urea can be purchased from Sinopharm Chemical Reagent Co., Ltd.; isoniazid Bases can be purchased from Aladdin Reagents Ltd.

Example 1: Preparation of dabigatran etexilate cocrystal by cooling crystallization method

Dabigatran etexilate and 7 co-crystal ligands (respectively p-hydroxybenzoic acid, 2,4-dihydroxybenzoic acid, succinic acid, nicotinamide, isonicotine, benzamide, and urea) were used in molar ratios. After mixing 1:1, dissolve in 10 ml methanol solvent, stir at 350 rpm for 6 hours, then cool down at 10°C/h until crystals precipitate, maintain for 3 hours, then filter with suction to obtain a sample, and dry the sample under reduced pressure to obtain the product.

Example 2: Preparation of dabigatran etexilate cocrystal by solvent method

Dabigatran etexilate and 7 co-crystal ligands (respectively p-hydroxybenzoic acid, 2,4-dihydroxybenzoic acid, succinic acid, nicotinamide, isonicotine, benzamide, and urea) were used in molar ratios. After mixing 1:1, dissolve in ethanol solvent, stir at 200 rpm for 3 hours, slowly add water dropwise to the solution, mix while adding, and then place at room temperature to slowly evaporate. When a large number of crystals are produced, suck out the remaining liquid and reduce the pressure. Drying gives the product.

Test Example 1: Equilibrium Solubility Test

Test method: Using water as the solvent, add an excess of dabigatran etexilate API and each product obtained in Example 1 and Example 2, and add a small amount of small glass beads. After sealing, shake at a constant temperature of 37°C and centrifuge. Clear, 0.45μm Filtrate through a microporous membrane, and the filtrate obtained is the test solution; in addition, accurately weigh about 5 mg of the dabigatran etexilate API as the reference substance, and dilute it to an appropriate concentration to form the reference substance solution. According to the high performance liquid chromatography method, the peak areas of the test solution and the reference solution were measured at 225 nm, and the content and equilibrium solubility were calculated by the external standard method.

Chromatographic conditions:

Detection wavelength: 225nm; chromatographic column: ZORBAX Eclipse XDB-C18 column (specification: 4.6mm×250mm, 5μm); mobile phase: acetonitrile-0.02mol/L ammonium acetate (triethylamine adjusts pH to 7.5) = 65:35 ;Flow rate: 1ml/min; Injection volume: 20μl; Column temperature: 30℃.

The equilibrium solubility results of dabigatran etexilate and each product obtained in Example 1 are shown in Figure 1. Among the eight co-crystal ligands, compared with the dabigatran etexilate API, 2,4-dihydroxybenzene The equilibrium solubility of the products with formic acid, p-hydroxybenzoic acid, and succinic acid as ligands was greatly improved, indicating that a eutectic was formed and the solubility was improved; while the remaining four ligands showed no significant improvement, indicating that no eutectic was formed. .

The equilibrium solubility results of each product obtained in Example 2 are shown in Figure 2, which is basically the same as the equilibrium solubility of each product obtained in Example 1, indicating that the products obtained by the two methods are basically the same.

Test Example 2: In vitro dissolution test

Test method: Use 900 ml of aqueous solution as the dissolution medium, add an appropriate amount of dabigatran etexilate and each product obtained in the above Example 1 and Example 2, and follow the dissolution determination method (2020 version of Chinese Pharmacopoeia General Chapter 0931 Second Method), 37℃, rotating speed is 75rpm, operate according to the law, take 5ml of the solution at 5, 10, 15, 20, 30, 45, 60, 90, 120, 180 and 240 minutes respectively, and filter it with a 0.45μm microporous membrane to obtain the test solution ; In addition, accurately weigh the dabigatran etexilate raw material medicinal dissolution medium and dilute it to an appropriate concentration to prepare a reference solution. According to the chromatographic conditions in Test Example 1, use high-performance liquid chromatography to measure the peak areas of the test solution and the reference solution at 225 nm, calculate the dissolution amounts at different time points using the external standard method, and draw a cumulative dissolution curve.

The in vitro dissolution curve of each product obtained in Example 1 is shown in Figure 2. The results show that the product with 2,4-dihydroxybenzoic acid, p-hydroxybenzoic acid, and succinic acid as ligands can be completely dissolved within 90 minutes (> 80%), indicating that the above three ligands and dabigatran etexilate all form a co-crystal, achieving the effect of improving in vitro dissolution; while the in vitro dissolution of dabigatran etexilate API and the products of the remaining four ligands The degrees are all less than <20%, indicating that no eutectic is formed.

The in vitro dissolution curve of each product obtained in Example 2 is shown in Figure 4, which is basically the same as the in vitro dissolution curve of each product obtained in Example 1.

Test Example 3: Characterization of dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal

For the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1 or Example 2, through X-ray Characterized by powder diffraction (XRPD), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR) and other methods.

a. Use a Bruker D8 Advance diffractometer (Bruker, Germany) to measure the powder diffraction pattern of the drug cocrystal obtained in Example 1. The test conditions are as follows: Cu, Kα, 40kV, 40mV as light source, step length 0.0128°, scanning range 3~ 45℃, room temperature. Under the same test conditions, the powder diffraction patterns of dabigatran etexilate, 2,4-dihydroxybenzoic acid, and dabigatran etexilate/2,4-dihydroxybenzoic acid physical mixture (molar ratio 1:1) were obtained .

Using Cu-Kα radiation, the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1 has characteristic peaks at 2θ (°) of 20.7°±0.2° and 24.9°±0.2°; There are also characteristic peaks at 17.9°±0.2°, 22.1°±0.2°, 23.2°±0.2°, and 24.1°±0.2°; there are also characteristic peaks at 21.9°±0.2°, 26.4°±0.2°, 27.1°±0.2°, There are characteristic peaks at 28.5°±0.2°; there are also characteristic peaks at 9.5°±0.2°, 11.0°±0.2°, 22.6°±0.2°, and 27.1°±0.2°; and at 17.8°±0.2° and 18.7°. ±0.2°, 25.7°±0.2°, 9.9°±0.2°, 19.1°±0.2°, 18.9°±0.2°, 20.3°±0.2°, 21.5°±0.2°, 17.0°±0.2°, 28.3°±0.2 There is a characteristic peak at °.

Specifically, the XRPD pattern of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1 is shown in Figure 5. The XRPD pattern of dabigatran etexilate API is shown in Figure 6.

Figure 7 shows the dabigatran etexilate/2,4-dihydroxybenzoic acid co-crystal prepared in Example 1 and dabigatran etexilate, 2,4-dihydroxybenzoic acid, and dabigatran etexilate/2, Comparative XRPD patterns of physical mixtures of 4-dihydroxybenzoic acid. Both dabigatran etexilate and 2,4-dihydroxybenzoic acid show obvious characteristic peaks, indicating that both have crystal structures; the characteristic peak of the dabigatran etexilate/2,4-dihydroxybenzoic acid physical mixture is dabigatran etexilate/2,4-dihydroxybenzoic acid. A simple superposition of the characteristic peaks of gatran etexilate and 2,4-dihydroxybenzoic acid shows that there is no interaction between the API and the ligand 2,4-dihydroxybenzoic acid in the physical mixture; compared with dabigatran etexilate API , the X-ray powder diffraction of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal of the present invention has characteristics at diffraction angles 2θ (error ±0.2 degrees) of 8.9°±0.2° and 15.5°±0.2°. The peak disappears, and there are characteristic peaks at 20.7°±0.2° and 24.9°±0.2°, indicating that a new structure is formed in the eutectic.

The XRPD pattern of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 2 is basically the same as Figure 5.

b. Use TA Q2000 differential scanning calorimeter (TA Instruments) to measure the drug eutectic obtained in Example 1. The test conditions are as follows: about 5 mg of the sample is packaged in an aluminum disk, the heating temperature is 25-300°C, and the heating rate is 10.0°C/ min, purge gas 50ml/min nitrogen, temperature calibration using NIST indium metal. Under the same test conditions, the DSC spectra of dabigatran etexilate and 2,4-dihydroxybenzoic acid were obtained.

Figure 8 shows the DSC spectrum of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1. picture 9 shows the DSC comparison spectra of dabigatran etexilate, 2,4-dihydroxybenzoic acid, and the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1. As shown in Figures 8 and 9, the melting points of dabigatran etexilate, 2,4-dihydroxybenzoic acid, and dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal are 134.4°C, 219.4°C, and 122.8℃. A change in melting point indicates the formation of a new phase.

The DSC spectrum of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 2 is basically the same as Figure 7.

c. Use a Nicolet 6700 Fourier transform infrared spectrometer (Thermo Fisher Scientific) to measure the drug cocrystal obtained in Example 1. The test conditions are as follows: use KBr dry tableting, and the scanning range is 4000 to 400 cm ^-1 .

Figure 10, Figure 11, and Figure 12 respectively show the FTIR of dabigatran etexilate, 2,4-dihydroxybenzoic acid, and the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1. Map. As shown in the figure, in the FTIR spectrum of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 1, the NH absorption peak of dabigatran etexilate is shifted from 3424.20cm ^-1 to 3412.05cm ^{- 1} , the C=O absorption peak shifts from 1729.21cm ^-1 to 1751.64cm ^-1 , indicating that the NH bond in dabigatran etexilate and the C=O bond in 2,4-dihydroxybenzoic acid may form or exist hydrogen bonds effect.

The FTIR spectrum of the dabigatran etexilate/2,4-dihydroxybenzoic acid cocrystal prepared in Example 2 is basically the same as Figure 12.

Test Example 4: In vivo pharmacokinetic test on beagle dogs

Test method: Take 9 male beagle dogs (purchased from Beijing Mars Biotechnology Co., Ltd.), weighing 9.5kg±1.0kg, and randomly divide them into three groups: test preparation group 1 (dabigatran etexilate/2,4 -Dihydroxybenzoic acid co-crystal group), test preparation group 2 (dabigatran etexilate/2,4-dihydroxybenzoic acid physical mixture group) and reference preparation group (dabigatran etexilate API group), 3 per group. Fast for 12 hours the day before the experiment and drink water freely. The two groups of beagle dogs were administered dabigatran etexilate at a dose equivalent to 150 mg. After administration, blood was collected from the leg vein at 10min, 30min, 1h, 1.5h, 2h, 3h, 4h, 6h, 8h, 12h, 24h, 36h, and 48h respectively. After collecting the blood, place it in a heparinized blood collection tube and centrifuge for 10 minutes. Precisely measure 100 μl of plasma and add it to the EP tube. Add 10 μl of verapamil internal standard solution (50ng/ml), mix well, add 0.4 ml of methanol, and vortex for 3 minutes. Centrifuge at 14000 r/min for 5 minutes; inject 5 μL of the supernatant, measure and calculate the blood drug concentration by LC-MS/MS, and calculate the pharmacokinetic parameters with DASS2.0 software.

a. Chromatographic conditions

Chromatographic column: Agilent narrow Bore RR SB-C18 column (specification: 2.1mm×100mm, 3.5μm); mobile phase: 0.1% formic acid aqueous solution (B): methanol (A), gradient elution: 0min90%B→0.5min10% B→2min10%B→2.01min 90%B; flow rate: 0.3ml/min; injection volume: 5μl; column temperature: 40°C.

b. Mass spectrometry conditions

Ion polarity: positive ions; ionization method: pneumatic-assisted electrospray ionization (ESI); ion detection method: multiple reaction monitoring (MRM); detection object: DE[M+H]+, m/z 628.1→289.0, Internal standard [M+H]+, m/z455.2→165.1; fragmentation voltages: 110V and 160V respectively; collision energy: DE 40eV, internal standard 30eV, drying gas flow rate: 10L/min; spray chamber pressure: 50psi; drying gas temperature: 350℃, capillary voltage: 4000V.

c. Pharmacokinetic data processing

The obtained blood drug concentration data were analyzed by DAS 2.0 analysis software.

d.Measurement results

Beagle dogs were orally administered dabigatran etexilate API, dabigatran etexilate/2,4-dihydroxybenzoic acid co-crystal prepared in Example 1, and dabigatran etexilate/2,4-dihydroxybenzoic acid. After physical mixture, the main pharmacokinetic parameters and relative bioavailability were calculated based on the average blood drug concentration (μg/mL) at different time points. The results are shown in Table 1 and Table 2.

Table 1: Main pharmacokinetic parameters of each preparation, n=3

Note: AUC _0→Tn and AUC _0→∞ are the areas under the plasma concentration curve at 0→Tn and 0→∞ times respectively, and C _max and T _max are the peak concentration and peak time respectively.

Table 2: Relative bioavailability results of test preparations, n=3

Note: Relative bioavailability = AUC _{test preparation} /AUC _{reference preparation} .

The above experimental results show that the test preparation (dabigatran etexilate/2,4-dihydroxybenzoic acid prepared in Example 1 cocrystal) and the reference preparation, the relative bioavailability was 164.02%. It can be seen that the dabigatran etexilate/2,4-dihydroxybenzoic acid drug co-crystal provided by the present invention can significantly improve the bioavailability of dabigatran etexilate, which illustrates the relationship between the active ingredient in the drug co-crystal and the co-crystal ligand. There is some interaction that enhances the absorption of the drug into the body.

In summary, compared with the prior art, the dabigatran etexilate cocrystal provided by the present invention has a simple prescription and imparts new physical and chemical properties to the active ingredient dabigatran etexilate without changing the molecular structure of the drug. The in vitro dissolution curve and the in vivo pharmacokinetics experimental results of beagle dogs show that the drug cocrystal provided by the present invention is superior to the dabigatran etexilate API and the physical mixture of the same proportion, and has the advantages of simple preparation process, low cost, and ease of use. It has the advantages of operation, stable quality, strong controllability, good reproducibility, and high bioavailability, and has potential application value.

Claims (13)

A co-crystal comprising dabigatran etexilate and a co-crystal ligand, the co-crystal ligand being selected from the group consisting of 2,4-dihydroxybenzoic acid, p-hydroxybenzoic acid, succinic acid, benzoic acid, tartaric acid, butyric acid Enedioic acid, fumaric acid or any combination thereof is more preferably 2,4-dihydroxybenzoic acid, p-hydroxybenzoic acid, and even more preferably 2,4-dihydroxybenzoic acid. The cocrystal of claim 1, wherein the molar ratio of dabigatran etexilate to the cocrystal ligand is 1:0.5~50, preferably 1:0.5~25, more preferably 1:0.5~10, further Preferably it is 1:0.5-5. The cocrystal of claim 1, wherein the cocrystal ligand is selected from the group consisting of 2,4-dihydroxybenzoic acid, p-hydroxybenzoic acid, and succinic acid; Preferably, the molar ratio of dabigatran etexilate to cocrystal ligand is 1:0.5~5, 1:0.8~4, 1:0.8~3, 1:0.8~2.5, 1:0.8~1.5, 1 :0.8～1.2, for example about 1:1. The co-crystal of claim 3, wherein the co-crystal ligand is 2,4-dihydroxybenzoic acid, and the molar ratio of dabigatran etexilate to the co-crystal ligand is about 1:1. The eutectic of claim 4, wherein in the X-ray powder diffraction pattern expressed in 2θ angle obtained using Cu-Kα radiation, the eutectic has characteristic peaks at 20.7°±0.2° and 24.9°±0.2°. ; Preferably, in the X-ray powder diffraction pattern expressed in 2θ angles obtained using Cu-Kα radiation, the eutectic is also at 17.9°±0.2°, 22.1°±0.2°, 23.2°±0.2°, 24.1°±0.2 There is a characteristic peak at °; Preferably, in the X-ray powder diffraction pattern expressed in 2θ angle obtained using Cu-Kα radiation, the eutectic is also at 21.9°±0.2°, 26.4°±0.2°, 27.1°±0.2°, 28.5°±0.2 There is a characteristic peak at °; Preferably, in the X-ray powder diffraction pattern expressed in 2θ angles obtained using Cu-Kα radiation, the eutectic is also at 9.5°±0.2°, 11.0°±0.2°, 22.6°±0.2°, 27.1°±0.2 There is a characteristic peak at °; Preferably, in the X-ray powder diffraction pattern expressed in 2θ angle obtained using Cu-Kα radiation, the eutectic is also at 17.8°±0.2°, 18.7°±0.2°, 25.7°±0.2°, 9.9°±0.2 There are characteristic peaks at 19.1°±0.2°, 18.9°±0.2°, 20.3°±0.2°, 21.5°±0.2°, 17.0°±0.2°, and 28.3°±0.2°; Preferably, the eutectic has substantially the same X-ray powder diffraction pattern as shown in Figure 5 using Cu-Kα radiation. The co-crystal of claim 4, wherein compared with the dabigatran etexilate API, in an X-ray powder diffraction pattern expressed at a 2θ angle obtained using Cu-Kα radiation, the co-crystal is at 8.9°±0.2° , there is no characteristic peak at 15.5°±0.2°, and there are characteristic peaks at 20.7°±0.2° and 24.9°±0.2°; Preferably, using Cu-Kα radiation, the dabigatran etexilate API has substantially the same X-ray as shown in Figure 6 Line powder diffraction pattern. The eutectic of claim 5 or 6, wherein the eutectic exhibits a melting endothermic phenomenon in the range of 120 to 130°C as measured by a differential scanning calorimeter, Preferably, the eutectic has substantially the same differential scanning calorimetry curve as shown in Figure 8. A pharmaceutical composition comprising the cocrystal according to any one of claims 1 to 7, and a pharmaceutically acceptable carrier or excipient. The method for preparing the cocrystal according to any one of claims 1 to 7 includes: a) Dissolve dabigatran etexilate and co-crystal ligand in the solvent; b) Cool down until crystals precipitate; c) optionally, isolating said crystals, optionally drying said crystals; Preferably, the solvent is selected from water, alcohol solvents, ester solvents, ketone solvents, ether solvents, nitrile solvents, alkane solvents, halogenated alkane solvents or any combination thereof; Preferably, the alcoholic solvent is selected from methanol, ethanol, isopropyl alcohol or any combination thereof; Preferably, the ester solvent is selected from methyl acetate, ethyl acetate, propyl acetate or any combination thereof; Preferably, the ketone solvent is selected from acetone, methyl isobutyl ketone or a combination thereof; Preferably, the ether solvent is selected from methyl tert-butyl ether, cyclopentyl methyl ether, tetrahydrofuran or any combination thereof; Preferably, the nitrile solvent is acetonitrile; Preferably, the alkane solvent is n-hexane; Preferably, the halogenated alkane solvent is methylene chloride; Preferably, the solvent is selected from methanol, ethanol, tert-butyl methyl ether, n-hexane, water or any combination thereof; The method for preparing the cocrystal according to any one of claims 1 to 7 includes: a) Dissolve dabigatran etexilate and co-crystal ligand in a non-aqueous solvent and stir; b) Slowly add water dropwise to the above solution and stir; c) Slowly volatilize to obtain a solid; d) optionally, drying the solid; Preferably, the solvent is selected from alcohol solvents, ester solvents, ketone solvents, ether solvents, nitrile solvents, alkane solvents, halogenated alkane solvents or any combination thereof; Preferably, the alcoholic solvent is selected from methanol, ethanol, isopropyl alcohol or any combination thereof; Preferably, the ester solvent is selected from methyl acetate, ethyl acetate, propyl acetate or any combination thereof; Preferably, the ketone solvent is selected from acetone, methyl isobutyl ketone or a combination thereof; Preferably, the ether solvent is selected from methyl tert-butyl ether, cyclopentyl methyl ether, tetrahydrofuran or any combination thereof; Preferably, the nitrile solvent is acetonitrile; Preferably, the alkane solvent is n-hexane; Preferably, the halogenated alkane solvent is methylene chloride; Preferably, the solvent is n-hexane. Use of the cocrystal according to any one of claims 1 to 7 in the preparation of anticoagulant drugs. The use of the cocrystal according to any one of claims 1 to 7 in the preparation of medicaments for the treatment and/or prevention of thrombosis. The cocrystal according to any one of claims 1 to 7 is used in the preparation of medicaments for the treatment and/or prevention of venous thromboembolism (VTE), deep vein thrombosis (DVT), pulmonary embolism (PE), stroke and systemic embolism. the use of.