Bezafibrate-Atenolol Crystal Complex and Its Preparation and Application

Abstract

The present invention relates to a bezafibrate - atenolol crystal complex and its preparation and application. Specifically, the present invention discloses a bezafibrate - atenolol crystal complex, which has significantly improved solubility of bezafibrate, significantly improved pharmacokinetic properties and significantly improved lipid - lowering effect. The present invention also discloses the preparation of the crystal complex and its application in the prevention and / or treatment of hyperlipidemia.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and specifically to a bezafibrate - atenolol crystal complex and its preparation and application. Background Art

[0002] Research shows that 61% of hypertensive patients have combined hyperlipidemia. The risk of cardiovascular diseases in such patients is higher than that in patients with only hypertension or hyperlipidemia. Controlling only blood pressure or blood lipids cannot effectively control the disease. At the same time, the number of administrations and doses of taking antihypertensive drugs and lipid - lowering drugs together lead to poor compliance of patients. Formulating antihypertensive drugs and lipid - lowering drugs into a single dosage form can not only improve the physicochemical properties of the drugs, produce good synergistic effects, but also facilitate the medication of such patients, increase the compliance of patients with medication, and has high social and economic significance.

[0003] A drug - drug multicomponent crystal complex is a crystal composed of two different drug active ingredients through non - covalent interactions such as hydrogen bonds, van der Waals forces, π…π stacking, etc., in a certain stoichiometric ratio. The formed drug - drug multicomponent crystal complex can not only improve the physicochemical properties of one or both APIs, such as solubility, dissolution rate, and stability, but also produce a synergistic effect, enhance the therapeutic effect, reduce the incidence of side effects, improve patient compliance, and at the same time reduce the costs required for the preparation and marketing of two separate drug products, and has more development prospects than compound preparations.

[0004] The chemical name of bezafibrate (BEZ): [p - (4 - chlorobenzoyl)- β -aminoethylphenoxy]isobutyric acid, with the chemical formula C 19 H 20 ClNO4, and the molecular weight is: 361.82. The chemical structural formula is as follows:

[0005] .

[0006] Bezafibrate is a lipid - regulating drug of the fenofibrate type. By activating peroxisome proliferator - activated receptors (PPARs), it promotes fatty acid oxidation and energy consumption, reduces cholesterol synthesis, and thus effectively reduces triglycerides and cholesterol in the blood. Bezafibrate has good lipid - lowering effects and is mainly used clinically to treat type I, II, III, IV, and V primary hyperlipidemias. In addition, bezafibrate also has the effect of inhibiting platelet aggregation, enhancing the effect of anticoagulant drugs, reducing plasma fibrinogen and plasma viscosity, and enhancing fibrinolytic activity, which helps prevent the occurrence and development of atherosclerosis. However, the water solubility of bezafibrate is poor, with a solubility in water of less than 0.01 mg / mL, which limits its oral absorption.

[0007] The chemical name of Atenolol (ATE): 4-[3-(2-hydroxy-3-isopropylamino)propoxy]benzeneacetamide, with the chemical formula C 14 H 22 N2O3, and the molecular weight is: 266.34. The chemical structural formula is as follows:

[0008] .

[0009] Atenolol is a highly selective β β-receptor blocker. By blocking the β1 β-receptors in the heart, it slows down the heart rate, delays atrioventricular conduction, and inhibits myocardial contraction, thereby reducing blood pressure and decreasing myocardial oxygen consumption. Clinically, in addition to being used to treat angina pectoris and arrhythmia, it is also used as a first-line drug for treating hypertension.

[0010] Therefore, the present invention designs and synthesizes a new lipid-lowering and blood-pressure-lowering drug - a drug multicomponent crystal complex to improve the physicochemical properties such as the solubility and dissolution rate of the drug, and further improve the absorption of the drug in the body, thereby enhancing the drug efficacy, providing a way for the combined treatment of hyperlipidemia and hypertension diseases. Summary of the Invention

[0011] The object of the present invention is to provide a bezafibrate-atenolol crystal complex and its preparation and application.

[0012] In the first aspect of the present invention, a bezafibrate-atenolol crystal complex is provided. The X-ray powder diffraction pattern of the crystal complex has characteristic peaks at 3 or more 2θ angles selected from the following group: 9.617±0.2°, 13.471±0.2°, 14.677±0.2°, 16.157±0.2°, 17.791±0.2°, 18.453±0.2°, 18.959±0.2°, 19.387±0.2°, 19.738±0.2°, 20.555±0.2°, 21.217±0.2°, 21.995±0.2°, 23.046±0.2°, 24.136±0.2°, 24.720±0.2°, 25.732±0.2°, 26.589±0.2°, 27.367±0.2°, 28.146±0.2°, 29.975±0.2°, 31.221±0.2°, 31.999±0.2°, 33.517±0.2°.

[0013] In another preferred example, the X-ray powder diffraction pattern of the crystal complex has characteristic peaks at 6 or more 2θ angles selected from the following group: 9.617 ± 0.2°, 13.471 ± 0.2°, 14.677 ± 0.2°, 16.157 ± 0.2°, 17.791 ± 0.2°, 18.453 ± 0.2°, 18.959 ± 0.2°, 19.387 ± 0.2°, 19.738 ± 0.2°, 20.555 ± 0.2°, 21.217 ± 0.2°, 21.995 ± 0.2°, 23.046 ± 0.2°, 24.136 ± 0.2°, 24.720 ± 0.2°, 25.732 ± 0.2°, 26.589 ± 0.2°, 27.367 ± 0.2°, 28.146 ± 0.2°, 29.975 ± 0.2°, 31.221 ± 0.2°, 31.999 ± 0.2°, 33.517 ± 0.2°.

[0014] In another preferred example, the X-ray powder diffraction pattern of the crystal complex has characteristic peaks at 10 or more 2θ angles selected from the following group: 9.617 ± 0.2°, 13.471 ± 0.2°, 14.677 ± 0.2°, 16.157 ± 0.2°, 17.791 ± 0.2°, 18.453 ± 0.2°, 18.959 ± 0.2°, 19.387 ± 0.2°, 19.738 ± 0.2°, 20.555 ± 0.2°, 21.217 ± 0.2°, 21.995 ± 0.2°, 23.046 ± 0.2°, 24.136 ± 0.2°, 24.720 ± 0.2°, 25.732 ± 0.2°, 26.589 ± 0.2°, 27.367 ± 0.2°, 28.146 ± 0.2°, 29.975 ± 0.2°, 31.221 ± 0.2°, 31.999 ± 0.2°, 33.517 ± 0.2°.

[0015] In another preferred example, the X-ray powder diffraction pattern of the crystal complex has characteristic peaks at the following 2θ angles: 9.617 ± 0.2°, 13.471 ± 0.2°, 14.677 ± 0.2°, 16.157 ± 0.2°, 17.791 ± 0.2°, 18.453 ± 0.2°, 18.959 ± 0.2°, 19.387 ± 0.2°, 19.738 ± 0.2°, 20.555 ± 0.2°, 21.217 ± 0.2°, 21.995 ± 0.2°, 23.046 ± 0.2°, 24.136 ± 0.2°, 24.720 ± 0.2°, 25.732 ± 0.2°, 26.589 ± 0.2°, 27.367 ± 0.2°, 28.146 ± 0.2°, 29.975 ± 0.2°, 31.221 ± 0.2°, 31.999 ± 0.2°, 33.517 ± 0.2°.

[0016] In another preferred example, the X-ray powder diffraction pattern of the crystal complex has characteristic peaks at the following 2θ angles: 9.617, 13.471, 14.677, 16.157, 17.791, 18.453, 18.959, 19.387, 19.738, 20.555, 21.217, 21.995, 23.046, 24.136, 24.720, 25.732, 26.589, 27.367, 28.146, 29.975, 31.221, 31.999, 33.517.

[0017] In another preferred example, in the crystal complex, the molar ratio of bezafibrate to atenolol is 1 - 3:1 - 3.

[0018] In another preferred example, in the crystal complex, the molar ratio of bezafibrate to atenolol is selected from the group consisting of: 3:1, 2:1, 1:1, 1:2, 1:3.

[0019] In another preferred example, in the crystal complex, the molar ratio of bezafibrate to atenolol is 1:1.

[0020] In another preferred example, the X-ray powder diffraction pattern of the crystal complex is substantially as Figure 1 shown.

[0021] In another preferred example, the crystal complex is a hydrate.

[0022] In another preferred example, in the crystal complex, the water content is 1 - 8 wt%, preferably 2 - 5 wt%, more preferably 3 - 4 wt%.

[0023] In another preferred example, the crystal complex has a weight loss of 2 - 5 wt% at 25 - 95 °C.

[0024] In another preferred example, the crystal complex has a weight loss of 3-4 wt% at 25-95 °C.

[0025] In another preferred example, the crystal complex has an endothermic peak at 120-130 °C.

[0026] In another preferred example, the DSC-TG diagram of the crystal complex is substantially as Figure 2 shown.

[0027] In another preferred example, the crystal complex has one or more characteristics selected from the following groups:

[0028] 1) The crystal complex belongs to the monoclinic system;

[0029] 2) The space group of the crystal complex is P space group 21 / c;

[0030] 3) In the crystal complex, the molar ratio of bezafibrate, atenolol and water is 1:1:1;

[0031] 4) The equilibrium solubility of bezafibrate in the buffer solution at pH 1.2 in the crystal complex is 0.001-0.002 mg / ml, preferably 0.0012-0.0018 mg / ml, more preferably 0.0013-0.0016 mg / ml;

[0032] 5) The equilibrium solubility of atenolol in the buffer solution at pH 1.2 in the crystal complex is 10-15 mg / ml, preferably 11-14 mg / ml, more preferably 12-13 mg / ml;

[0033] 6) The equilibrium solubility of bezafibrate in the buffer solution at pH 6.8 in the crystal complex is 10-15 mg / ml, preferably 11-14 mg / ml, more preferably 12-13 mg / ml;

[0034] 7) The equilibrium solubility of atenolol in the buffer solution at pH 6.8 in the crystal complex is 8-15 mg / ml, preferably 10-12 mg / ml, more preferably 10-ll mg / ml.

[0035] In the second aspect of the present invention, a preparation method of the bezafibrate-atenolol crystal complex described in the first aspect of the present invention is provided, including the following steps:

[0036] 1) Provide bezafibrate, atenolol and a first solvent;

[0037] 2) Add the bezafibrate and the atenolol to the first solvent, and stir at 10 - 40 °C for 10 - 36 h to obtain a first mixture;

[0038] 3) Separate the solid and liquid of the first mixture, collect the solid precipitate, and dry it at 30 - 50 °C for 6 - 18 h to obtain the bezafibrate-atenolol crystal complex.

[0039] In another preferred example, in step 2), the molar ratio of the added bezafibrate to the atenolol is 1 - 3:1 - 3, preferably 1 - 2:1 - 2, and most preferably 1:1.

[0040] In another preferred example, in step 2), the mass-volume ratio of the bezafibrate to the first solvent is 150 - 250 mg:3 - 5 mL, preferably 180 - 220 mg:3 - 5 mL, more preferably 180 - 220 mg:4 mL, and most preferably 200 mg:4 mL.

[0041] In another preferred example, in step 2), the mass-volume ratio of the atenolol to the first solvent is 100 - 200 mg:3 - 5 mL, preferably 130 - 170 mg:3 - 5 mL, more preferably 140 - 160 mg:3 - 5 mL, and most preferably 140 - 160 mg:4 mL.

[0042] In another preferred example, the first solvent is selected from the group consisting of water, acetonitrile, ethyl acetate, methanol, ethanol, n-propanol, isopropanol, tetrahydrofuran, acetone, isopropyl ether, dichloromethane, methyl tert-butyl ether, or a combination thereof.

[0043] In another preferred example, the first solvent is water.

[0044] In another preferred example, in step 2), the stirring is carried out at 15 - 35 °C, preferably 20 - 30 °C.

[0045] In another preferred example, in step 2), the stirring is carried out for 15 - 30 h, preferably 20 - 26 h, more preferably 22 - 25 h.

[0046] In another preferred example, in step 3), the temperature of the drying treatment is 35 - 45 °C, preferably 40 °C.

[0047] In another preferred example, in step 3), the treatment time of the drying treatment is 10 - 15 h, preferably 12 h.

[0048] In another preferred example, in step 3), the drying is carried out in a vacuum drying oven.

[0049] In a third aspect of the present invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a safe and effective amount of the bezafibrate - atenolol crystal complex described in the first aspect of the present invention.

[0050] In a fourth aspect of the present invention, there is provided a use of the bezafibrate - atenolol crystal complex described in the first aspect of the present invention for the preparation of a drug for preventing and / or treating cardiovascular and cerebrovascular system diseases.

[0051] In another preferred embodiment, the cardiovascular and cerebrovascular system diseases are selected from the group consisting of: hypertension, hyperlipidemia, or a combination thereof.

[0052] In another preferred embodiment, the cardiovascular and cerebrovascular system disease is hyperlipidemia combined with hypertension.

[0053] It should be understood that within the scope of the present invention, the above - mentioned technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 It is the X - ray powder diffraction (PXRD) spectrum of the bezafibrate - atenolol crystal complex prepared in Example 1.

[0055] Figure 2 It is the differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) spectra of the bezafibrate - atenolol crystal complex prepared in Example 1.

[0056] Figure 3 It is the unit cell diagram of the single - crystal X - ray diffraction (SCXRD) of the bezafibrate - atenolol crystal complex prepared in Example 1.

[0057] Figure 4 It is the equilibrium solubility diagram of bezafibrate and the crystal complex in 37 °C hydrochloric acid buffer solution (pH 1.2) and phosphate - buffered saline solution (pH 6.8) in Example 3.

[0058] Figure 5 It is the equilibrium solubility diagram of atenolol and the crystal complex in 37 °C hydrochloric acid buffer solution (pH 1.2) and phosphate - buffered saline solution (pH 6.8) in Example 3.

[0059] Figure 6 It is the powder dissolution curve spectrum of bezafibrate and the crystal complex in 37 °C phosphate - buffered saline solution (pH 6.8) for 6 hours in Example 4.

[0060] Figure 7Powder dissolution curve spectrogram of atenolol and crystal complex in phosphate buffer solution (pH 6.8) at 37 °C for 6 hours in Example 4.

[0061] Figure 8 Pharmacokinetic curve of bezafibrate in SD rats of bezafibrate-atenolol crystal complex in Example 1.

[0062] Figure 9 Pharmacokinetic curve of atenolol in SD rats of bezafibrate-atenolol crystal complex in Example 1. Detailed implementation manners

[0063] After long-term and in-depth research, the present inventors unexpectedly prepared a bezafibrate-atenolol crystal complex, which has excellent solubility of bezafibrate and excellent pharmacokinetic properties and is very suitable for combination drug use. On this basis, the inventors completed the present invention.

[0064] Terms

[0065] When referring to spectrograms or / and data appearing in figures, "peak" refers to a feature that can be recognized by those skilled in the art and does not belong to background noise.

[0066] The present invention relates to the bezafibrate and atenolol crystal complexes, which exist in a substantially pure crystalline form.

[0067] "Substantially pure" means that a crystal form is substantially free of one or more other crystal forms, that is, the purity of the crystal form is at least 80%, or at least 85%, or at least 90%, or at least 93%, or at least 95%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9%, or the crystal form contains other crystal forms, and the percentage of the other crystal forms in the total volume or total weight of the crystal form is less than 20%, or less than 10%, or less than 5%, or less than 3%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01%.

[0068] "Substantially free of" means that the percentage of one or more other crystal forms in the total volume or total weight of the crystal form is less than 20%, or less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1%, or less than 0.5%, or less than 0.1%, or less than 0.01%.

[0069] "Relative intensity" (or "relative peak height") in the X-ray powder diffraction pattern refers to the ratio of the intensity of other peaks to the intensity of the first strong peak when the intensity of the first strong peak among all diffraction peaks of the X-ray powder diffraction pattern (PXRD) is 100%.

[0070] In the present invention, "room temperature" refers to a temperature ranging from approximately 10 °C to approximately 40 °C. In some embodiments, "room temperature" refers to a temperature ranging from approximately 20 °C to approximately 30 °C; in other embodiments, "room temperature" refers to 20 °C, 22.5 °C, 25 °C, 27.5 °C, and so on.

[0071] Bezafibrate-atenolol crystal complex

[0072] The present invention provides a bezafibrate-atenolol crystal complex, and the X-ray powder diffraction pattern of the crystal complex has the characteristics as described above.

[0073] Pharmaceutical composition

[0074] The present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a safe and effective amount of the bezafibrate-atenolol crystal complex.

[0075] The pharmaceutical composition of the present invention comprises the crystal complex of the present invention within a safe and effective amount range and a pharmacologically acceptable excipient or carrier. The "safe and effective amount" herein refers to: an amount of the crystal complex sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1 - 2000 mg of the crystal complex of the present invention per dose, more preferably, contains 10 - 1000 mg of the crystal complex of the present invention per dose. Preferably, the "per dose" is a capsule or a tablet.

[0076] The "pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" herein means that the components in the composition can be mixed with the crystal complex of the present invention and with each other without significantly reducing the efficacy of the crystal complex. Some examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween®), wetting agents (such as sodium dodecyl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0077] The pharmaceutical composition is an injection, a capsule, a tablet, a pill, a powder, or a granule.

[0078] There is no particular limitation on the administration mode of the pharmaceutical composition of the present invention. Representative administration modes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0079] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active crystalline complex is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or calcium phosphate, or with the following components: (a) fillers or bulking agents, e.g., starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, e.g., hydroxypropylmethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, e.g., glycerin; (d) disintegrants, e.g., agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, e.g., paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (h) adsorbents, e.g., kaolin; and (i) lubricants, e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0080] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and release of the active compound or compounds in such compositions can be delayed and released in a particular part of the digestive tract. Examples of embedding components that can be used are polymeric and wax-like substances. Optionally, the active compound can also be in the form of microcapsules with one or more of the above excipients.

[0081] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, the liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers, and emulsifying agents, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil or mixtures of these substances, etc.

[0082] In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, and perfumes.

[0083] In addition to the active compound, the suspension may contain suspending agents, e.g., ethoxylated isostearyl alcohols, polyoxyethylene sorbitol, and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, and agar, or mixtures of these substances, etc.

[0084] Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or non-aqueous solution, dispersion, suspension or emulsion, and sterile powders for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0085] Dosage forms of the pharmaceutical composition of the present invention for topical administration include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required, if necessary.

[0086] The pharmaceutical composition of the present invention can be administered alone or in combination with other pharmaceutically acceptable drugs.

[0087] The treatment method of the present invention can be administered alone or in combination with other treatment means or therapeutic drugs.

[0088] When using the pharmaceutical composition, a safe and effective amount of the crystalline complex of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is an effective dosage considered pharmaceutically. For a person weighing 60 kg, the daily dosage is usually 1 - 2000 mg, preferably 50 - 1000 mg. Of course, the specific dosage should also consider factors such as the route of administration and the health condition of the patient, which are within the scope of the skills of a skilled physician.

[0089] Compared with the prior art, the present invention has the following main advantages:

[0090] (1) The bezafibrate - atenolol crystalline complex can significantly improve the solubility of bezafibrate;

[0091] (2) The bezafibrate - atenolol crystalline complex has excellent pharmacokinetic properties, has a synergistic effect, and is very suitable for combination use;

[0092] (3) The bezafibrate - atenolol crystalline complex has a significantly enhanced lipid - lowering effect and can restore the TC / TG level to the normal level faster and better;

[0093] (4) The preparation method of the bezafibrate - atenolol crystalline complex is simple, has good repeatability, and is suitable for industrial production.

[0094] The present invention will be further illustrated below with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0095] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to persons skilled in the art. In addition, any methods and materials similar or equivalent to the described content can be applied to the methods of the present invention. The preferred methods and materials described herein are for illustrative purposes only.

[0096] It should be understood that the raw materials and reagents used in the examples are all commercially available products, and the reagents, instruments or operation steps not described herein are all contents that can be routinely determined by those of ordinary skill in the art.

[0097] General test methods

[0098] X-ray powder diffraction (PXRD) can detect information such as changes in crystal form, crystallinity, and crystal structure state, and is a commonly used means for identifying crystal forms. The peak positions of the X-ray powder diffraction pattern mainly depend on the crystal form structure and are relatively insensitive to experimental details, while the relative peak heights depend on many factors related to sample preparation and instrument geometry. Therefore, in some embodiments, the crystal complexes of the present invention are characterized by having an X-ray powder diffraction pattern with certain peaks, which is substantially as shown in the X-ray powder diffraction pattern provided in the accompanying drawings of the present invention. At the same time, the measurement of 2 θ of the X-ray powder diffraction pattern can have experimental errors, and there may be slight differences in the measurement of 2 θ of the X-ray powder diffraction pattern between different instruments and different samples. Therefore, the numerical value of the 2 θ cannot be regarded as absolute. According to the instrument conditions used in this experiment, there is an error tolerance of ±0.2° for the diffraction peaks.

[0099] Differential scanning calorimetry (DSC) is a technique that measures the energy difference between a sample and an inert reference (commonly used α -Al2O3) as a function of temperature under programmed control by continuously heating or cooling. The height of the endothermic peak of the differential scanning calorimetry spectrum curve depends on many factors related to sample preparation and instrument geometry, and the peak position is relatively insensitive to experimental details. Therefore, in some embodiments, the crystal complexes of the present invention are characterized by having a differential scanning calorimetry graph with characteristic peak positions. At the same time, the differential scanning calorimetry spectrum can have experimental errors, and there may be slight differences in the peak positions and peak values of the differential scanning calorimetry spectrum between different instruments and different samples. Therefore, the numerical values of the peak positions or peak values of the endothermic peaks of the differential scanning calorimetry spectrum cannot be regarded as absolute. According to the instrument conditions used in this experiment, there is an error tolerance of ±3 °C for the endothermic peaks.

[0100] Thermogravimetric analysis is a technique for measuring the mass change of a substance with temperature under programmed control. It is applicable to examining the loss of solvents in crystals or the processes of sample sublimation and decomposition, and can infer the situation of crystal water or crystal solvents contained in the crystal. The mass change shown by the thermogravimetric analysis curve depends on many factors such as sample preparation and instruments. There are slight differences in the mass changes detected by thermogravimetric analysis between different instruments and different samples. According to the instrument conditions used in this experiment, there is an error tolerance of ±0.5 for the mass change.

[0101] In the context of the present invention, the 2 θ values in the X-ray powder diffraction pattern are all in degrees (°).

[0102] Unless otherwise specified in the parameters, all the following analyses are carried out at room temperature.

[0103] For X-ray powder diffraction (PXRD) of the crystal complexes in the examples, a powder diffractometer was used. The instrument was irradiated with a Cu target (40 kV, 40 mA) and carried out at room temperature using a D / texUltra detector. The scanning range was from 3° to 45° in the 2 θ interval, and the scanning speed was 20 ° / min.

[0104] Differential scanning calorimetry (DSC) analysis was performed on the crystal complexes in the examples. The operation and analysis steps are as follows: A TA Q2000 differential scanning calorimeter was used, with an N2 atmosphere and a heating rate of 10 °C / min. In the differential scanning calorimetry graph, the abscissa represents temperature (°C), and the ordinate represents the heat flow released per unit mass of the substance (W / g).

[0105] Thermogravimetric (TGA) analysis was performed on the crystal complexes in the examples. The operation and analysis steps are as follows: A TAQ500 thermogravimetric analyzer was used, with an N2 atmosphere and a heating rate of 10 °C / min. In the TGA graph, the abscissa represents temperature, and the ordinate represents mass percentage (%).

[0106] For single crystal X-ray diffraction (SCXRD) analysis of the examples, the operation and analysis steps are as follows: A Bruker D8Venture (Bruker, Karlsruhe, Germany) diffractometer was used. Diffraction data were collected using a CCD under Cu-K α radiation ( λ =0.83 Å), the data were integrated and reduced using APEX3 software, the structure was solved by the direct method using OLEX2 software, and refined by full matrix least squares using the SHELXL program.

[0107] The dissolution analysis of the crystal complex in the examples was carried out with the following operation and analysis steps: An Agilent 1260 series high performance liquid chromatograph was used, column temperature: 40 °C; injection volume: 20 μ L; detection wavelength: 226 nm; flow rate: 1 mL / min; the retention time of bezafibrate compound was 7.14 min; the retention time of atenolol compound was 5.51 min, mobile phase: phase A was KH2PO4 aqueous solution, phase B was methanol, A:B = 30:70 (bezafibrate), A:B = 60:40 (atenolol).

[0108] Analysis of blood drug concentration in SD rats was carried out with the following operation and analysis steps: Q Exactive LC-MS was used, and the mass spectrometry parameters were: ion spray voltage 3.50 kV, gas temperature 350 °C, gas flow rate 10 L / min, sheath gas flow rate 35 L / min. The chromatographic conditions were: Waters C18 chromatographic column (1.7 μ m, 2.1 mm×100 mm), flow rate was 0.35 mL / min, column temperature was 40 °C, the mobile phase was composed of (A) 0.1% formic acid aqueous solution and (B) acetonitrile solution, and the gradient elution program was: 0 - 9 min: 5 - 99% B; 9 - 12 min: 99% B; 12 - 12.1 min: 99 - 5% B; 12.1 - 15 min: 5% B, the injection volume was set to 10 μ L, the retention time of atenolol was 2.17 min, and the retention time of bezafibrate was 5.99 min.

[0109] Determination of TC and TG contents in SD rat serum was carried out with the following operation and analysis steps: The serum was thawed at room temperature. After adding the corresponding volumes of distilled water / standard / sample and working solution according to the following operation table, the 96-well plate was shaken to mix evenly, After incubation at 37 °C for 10 min, the absorbance value of each well at 500 nm was measured by an enzyme-labeled instrument. The TC and TG contents were calculated by the following formula:

[0110]

[0111] Preparation and characterization of bezafibrate-atenolol crystal complex in Example 1

[0112] Equimolar amounts of bezafibrate (200 mg, 0.55 mmol) and atenolol (147.23 mg, 0.55 mmol) were added to 4 mL of water. After stirring at room temperature for 24 h, solid-liquid separation was carried out. The lower layer of solid precipitate was placed in a vacuum drying oven at 40 °C for 12 h. The solid powder collected was the bezafibrate-atenolol crystal complex, and the product mass was 250.03 mg.

[0113] Figure 1 The X-ray powder diffraction (PXRD) pattern of the bezafibrate-atlmolol crystal complex prepared in Example 1. The specific data are shown in Table 1.

[0114] Table 1

[0115]

[0116] Figure 2 The differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) patterns of the bezafibrate-atlmolol crystal complex prepared in Example 1.

[0117] From Figure 2 it can be seen that the bezafibrate-atlmolol crystal complex shows a weight loss of 3.6% in the range of 25 - 95 °C, which is attributed to the desolvation phenomenon, indicating that the crystal complex is a hydrate. At the same time, its differential scanning calorimetry pattern shows an endothermic peak at 89.08 °C, which is the desolvation peak, and an endothermic peak at 126.50 °C, which is attributed to the melting peak.

[0118] Single crystal X-ray diffraction of the bezafibrate-atlmolol crystal complex in Example 2

[0119] Single crystal X-ray diffraction analysis was performed on the bezafibrate-atlmolol crystal complex, and the results are as Figure 3 shown.

[0120] Figure 3 The unit cell diagram of the single crystal X-ray diffraction (SCXRD) of the bezafibrate-atlmolol crystal complex prepared in Example 1.

[0121] From Figure 3 it can be seen that the bezafibrate-atlmolol crystal belongs to the monoclinic system, P space group P21 / c, and its unit cell contains 4 bezafibrate cations, 4 atlmolol anions and 4 water molecules.

[0122] The crystal structure parameters of bezafibrate-atlmolol are shown in Table 2.

[0123] Table 2

[0124]

[0125] Equilibrium solubility experiment of the bezafibrate-atlmolol crystal complex in Example 3

[0126] To study the equilibrium solubility of bezafibrate - atenolol in hydrochloric acid buffer (pH 1.2) and phosphate buffered saline (pH 6.8), and analyze the dissolution ability in the simulated gastrointestinal fluid environment at 37 °C, the specific experimental method is as follows: Excessive sample powders were added to 1 mL of pH 1.2 and pH 6.8 buffer solutions respectively. After shaking in a constant temperature shaker at 37 °C for 48 h, centrifugation was carried out, and the supernatant was filtered through a 0.22 μ μm cellulose ester membrane filter, and the concentrations of bezafibrate and atenolol were determined by high - performance liquid chromatography.

[0127] Figure 4 Equilibrium solubility diagram of bezafibrate and crystal complex in hydrochloric acid buffer (pH 1.2) and phosphate buffered saline (pH 6.8) at 37 °C for Example 3.

[0128] Figure 5 Equilibrium solubility diagram of atenolol and crystal complex in hydrochloric acid buffer (pH 1.2) and phosphate buffered saline (pH 6.8) at 37 °C for Example 3.

[0129] The equilibrium solubility results of bezafibrate, atenolol and bezafibrate - atenolol crystal complex in pH 1.2 buffer and pH 6.8 buffer are shown in Table 3.

[0130] Table 3

[0131]

[0132] As can be seen from Table 3, in the simulated gastric fluid environment of pH 1.2 buffer, the equilibrium solubility of bezafibrate in bezafibrate - atenolol increased by a factor of 1.12, and the equilibrium solubility of atenolol decreased. In the pH environment of simulated intestinal fluid (pH 6.8 buffer), the equilibrium solubility of bezafibrate in bezafibrate - atenolol increased by a factor of 3.07. Therefore, bezafibrate - atenolol increases the solubility of bezafibrate, which is beneficial to promoting the absorption of bezafibrate in the gastrointestinal tract.

[0133] Powder dissolution experiment of bezafibrate - atenolol crystal complex in Example 4

[0134] To study the powder dissolution behavior of bezafibrate, atenolol, and the bezafibrate-atenolol crystal complex in phosphate buffer solution (pH 6.8) and analyze the dissolution rate at 37 °C, the specific experimental method is as follows: Excessive powders of bezafibrate, atenolol, and the bezafibrate-atenolol crystal complex were respectively added to glass bottles containing the dissolution medium of pH 6.8 buffer solution. The water bath temperature was set at 37 ± 0.5 °C, and the stirring speed was 200 rpm. Samples were taken at preset time points (1, 3, 5, 7, 10, 15, 20, 30, 45, 60, 90, 120, 180, 240, 360 min). After taking 600 μ mL of the supernatant, an equal volume of fresh dissolution medium was immediately replenished. The supernatant was immediately filtered through a 0.22 μ μm filter, diluted, and the concentration was measured by high performance liquid chromatography.

[0135] Figure 6 Powder dissolution curve spectra of bezafibrate and the crystal complex in phosphate buffer solution (pH 6.8) at 37 °C for 6 hours in Example 4.

[0136] Figure 7 Powder dissolution curve spectra of atenolol and the crystal complex in phosphate buffer solution (pH 6.8) at 37 °C for 6 hours in Example 4.

[0137] The powder dissolution results of bezafibrate, atenolol, and the bezafibrate-atenolol crystal complex in pH 6.8 buffer solution are as follows:

[0138] In pH 6.8 buffer solution, the bezafibrate powder gradually dissolved, reached equilibrium after 120 min, and maintained at the highest concentration. The solubility of bezafibrate was approximately 2.42 mg / mL.

[0139] In pH 6.8 buffer solution, the bezafibrate-atenolol crystal complex powder gradually dissolved. After 120 min, the solution concentration of bezafibrate reached equilibrium and maintained at the highest concentration. The solubility of bezafibrate was approximately 9.83 mg / mL.

[0140] In pH 6.8 buffer solution, the atenolol powder gradually dissolved, reached equilibrium after 120 min, and maintained at the highest concentration. The solubility of atenolol was approximately 23.13 mg / mL.

[0141] In pH 6.8 buffer solution, the bezafibrate-atenolol crystal complex powder gradually dissolved. After 120 min, the solution concentration of atenolol reached equilibrium and maintained at the highest concentration. The solubility of atenolol was approximately 8.17 mg / mL.

[0142] In summary, for the dissolution of the powder in phosphate buffer solution (pH 6.8), it was found that the dissolution rate of bezafibrate - atenolol was higher than that of bezafibrate and slightly lower than that of atenolol. At the same time, the maximum solubility of bezafibrate was increased, and the maximum solubility of atenolol was slightly decreased, indicating that bezafibrate - atenolol had a good promoting effect on the dissolution rate and solubility of bezafibrate, and slightly decreased the dissolution rate and solubility of atenolol.

[0143] Example 5 Pharmacokinetic Experiment of Bezafibrate - Atenolol Crystal Complex in SD Rats

[0144] In order to study the maximum plasma drug concentration of the crystal complex in SD rats, a pharmacokinetic experiment was carried out, and the specific experimental method was as follows:

[0145] The rats (female rats) were randomly divided into 4 groups, with 3 rats in each group. They were fasted for 12 h before the experiment and allowed free access to water.

[0146] Grouping: BEZ (bezafibrate), ATE (atenolol), BEZ - ATE (bezafibrate - atenolol crystal complex), PM (physical mixture of bezafibrate and atenolol)

[0147] Administration method: Administration by suspension

[0148] Administration dose: Calculated based on the rat mass of 200 g / rat, the administration doses are shown in Table 4.

[0149] Table 4

[0150]

[0151] Experimental operation: The 4 groups were respectively given BEZ, ATE, BEZ - ATE, and PM by gavage. Blood samples of about 200 μ μL were collected from the orbital cavity at 10 min, 20 min, 30 min, 40 min; 1 h, 2 h, 4 h, 6 h, 9 h, and 11 h after administration. The plasma was separated by centrifugation at 3000 rpm for 10 min and stored at -20 °C. The plasma drug concentration was determined by LC - MS.

[0152] Figure 8 For the bezafibrate plasma concentration - time curve of the bezafibrate - atenolol crystal complex in SD rats in Example 1.

[0153] Figure 9 For the atenolol plasma concentration - time curve of the bezafibrate - atenolol crystal complex in SD rats in Example 1.

[0154] As Figure 8 and Figure 9As shown, the results indicate that after single-agent BEZ, BEZ-ATE crystal complex, and PM were administered by gavage, the drug BEZ in SD rats C max was 1.83 μ g / mL, 12.15 μ g / mL, and 9.04 μ g / mL, respectively. After BEZ-ATE administration, the C max amount of BEZ in blood was 6.64 times that of single-agent BEZ and 1.34 times that of PM. The area under the concentration-time curve ( AUC 0-660min ) was 11.77 times that of BEZ and 2.33 times that of PM. It can be seen that after the formation of the crystal complex of BEZ and ATE and administration, compared with single-agent BEZ and physical mixture PM, there was higher C max and AUC in SD rats, which was more conducive to exerting the drug effect.

[0155] For ATE, BEZ-ATE, and PM after gavage administration, the drug ATE in SD rats C max was 1.67 μ g / mL, 1.32 μ g / mL, and 1.61 μ g / mL, respectively. After BEZ-ATE administration, the C max was slightly lower than that of ATE and PM. The area under the concentration-time curve ( AUC 0-660min ) was 1.17 times that of ATE and 1.03 times that of PM. It can be seen that after the formation of the crystal complex of ATE and BEZ, compared with single-agent ATE and physical mixture PM, C max it was slightly decreased, but had higher AUC .

[0156] Therefore, the BEZ-ATE crystal complex improved the pharmacokinetic parameters of both BEZ and ATE, which was beneficial to achieving an improvement in the drug effect.

[0157] Example 6 Evaluation Experiment on the Hypolipidemic Effect of Bezafibrate-Atenolol Crystal Complex in SD Rats

[0158] To study the efficacy of the crystal complex on hyperlipidemia, an evaluation experiment on the hypolipidemic effect was carried out in SD rats. The specific experimental method is as follows:

[0159] Male rats were acclimated for 5 days and then divided into a control group and a modeling group (the modeling group was further divided into a model control group, an ATE group, a BEZ group, and a BEZ-ATE group). The control group was fed a standard diet, while the modeling group was fed a high-fat diet. After 8 consecutive weeks of feeding, blood was collected from the orbital cavity, and serum was separated for determination of cholesterol (TC) and triglyceride (TG) levels. After successful modeling, the drug was suspended in 0.5% CMC-Na and administered. The control group and the modeling control group were gavaged with an equal volume of drug-free 0.5% CMC-Na. The doses of BEZ, ATE, and BEZ-ATE were 18 mg / kg, 13.25 mg / kg, and 32.59 mg / kg, respectively. Orbital blood was collected after 1 and 2 weeks of continuous administration, and serum cholesterol (TC) and triglyceride (TG) levels were determined.

[0160] The results are shown in Table 5.

[0161] Table 5

[0162]

[0163] As shown in Table 5, TC levels did not decrease significantly in the blank and model control groups after one week of administration, but decreased slightly in the ATE group. TC levels decreased significantly in the BEZ and BEZ-ATE groups, with the degree of reduction in BEZ-ATE being greater than in BEZ. After two weeks of administration, TC levels in the BEZ group decreased further, while no significant change was observed in the BEZ-ATE group. This indicates that TC levels in hyperlipidemic rats returned to normal after one week of administration. This suggests that the BEZ-ATE crystal complex has a more rapid and effective effect on reducing TC levels than BEZ.

[0164] Regarding TG levels, after one week of administration, there was no significant decrease in TG levels in the blank, model, and ATE groups. However, there was a significant decrease in TG levels in the BEZ and BEZ-ATE groups, with the decrease in the BEZ-ATE group being greater than that in the BEZ group. After two weeks of administration, TG levels in the BEZ group further decreased, while there was no significant change in the BEZ-ATE group. This suggests that TG levels in hyperlipidemic rats returned to normal after one week of administration. This suggests that the BEZ-ATE crystal complex has a faster and more effective effect on lowering TG levels than BEZ.

[0165] Therefore, after bezafibrate and atenolol form a crystal complex, they can exert a faster and better lipid-lowering effect.

[0166] All documents mentioned in this invention are cited herein by reference as if each individual document was cited by reference. In addition, it should be understood that after reading the above teachings of this invention, those skilled in the art can make various changes or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. Bezafibrate-Atenolol Crystal Complex, characterized in that, The X-ray powder diffraction pattern of the crystal complex has characteristic peaks at the following 2θ angles: 9.617±0.2°, 13.471±0.2°, 14.677±0.2°, 16.157±0.2°, 17.791±0.2°, 18.453±0.2°, 18.959±0.2°, 19.387±0.2°, 19.738±0.2°, 20.555±0.2°, 21.217±0.2°, 21.995±0.2°, 23.046±0.2°, 24.136±0.2°, 24.720±0.2°, 25.732±0.2°, 26.589±0.2°, 27.367±0.2°, 28.146±0.2°, 29.975±0.2°, 31.221±0.2°, 31.999±0.2°, 33.517±0.2°; The crystal complex is a hydrate; In the crystal complex, the molar ratio of bezafibrate, atenolol and water is 1:1:

1.

2. The crystalline complex according to claim 1, wherein The X-ray powder diffraction pattern of the crystal complex is basically as shown in Figure 1.

3. The crystal complex according to claim 1, wherein The X-ray powder diffraction pattern of the crystal complex has characteristic peaks at the following 2θ angles: 9.617°, 13.471°, 14.677°, 16.157°, 17.791°, 18.453°, 18.959°, 19.387°, 19.738°, 20.555°, 21.217°, 21.995°, 23.046°, 24.136°, 24.720°, 25.732°, 26.589°, 27.367°, 28.146°, 29.975°, 31.221°, 31.999°, 33.517°.

4. The crystalline complex according to claim 1, wherein The crystal complex has a weight loss of 2-5 wt% at 25-95 °C.

5. The crystal complex according to claim 1, wherein The crystal complex has an endothermic peak at 120-130 °C.

6. A method for preparing the bezafibrate - atenolol crystal complex according to claim 1, characterized in that, Comprising the following steps: 1) Provide bezafibrate, atenolol and a first solvent; 2) Add the bezafibrate and the atenolol to the first solvent, and stir at 10-40 °C for 10-36 h to obtain a first mixture; 3) Separate the solid and liquid of the first mixture, collect the solid precipitate, and dry it at 30-50 °C for 6-18 h to obtain the bezafibrate-atenolol crystal complex; The first solvent is water.

7. A pharmaceutical composition, characterized in that, Comprising a pharmaceutically acceptable carrier and a safe and effective amount of the bezafibrate-atenolol crystal complex according to claim 1.

8. Use of the bezafibrate - atenolol crystal complex according to claim 1, characterized in that, For the preparation of a drug for preventing and / or treating cardiovascular and cerebrovascular system diseases; The cardiovascular and cerebrovascular system diseases are selected from the group consisting of: hypertension, hyperlipidemia, or a combination thereof.

9. The use according to claim 8, wherein, The cardiovascular and cerebrovascular system disease is hyperlipidemia combined with hypertension.

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