Mylrigol-cinnamic acid crystals and a method of preparation

Abstract

The application belongs to the technical field of pharmaceutical chemistry, and provides a high-purity milrinone-cinnamic acid crystal, a preparation method thereof is simple in operation, a crystallization process is easy to control, and reproducibility is good. The prepared milrinone-cinnamic acid crystal has significantly enhanced stability and solubility compared with free base and crystal forms thereof, thereby being beneficial to storage, transportation and application in preparation of a preparation of a product, and having enhanced bioavailability and absorption performance.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and particularly relates to a milrinone-cinnamic acid crystal and a preparation method thereof. BACKGROUND

[0002] Milrinone, with a chemical name of 1,6-dihydro-2-methyl-6-oxo-[3,4-bipyridine]-5- carbonitrile, a molecular formula of C 12 H9N3O, a molecular weight of 211.22, and a structure formula of:

[0003]

[0004] Milrinone was first developed by the American Sterling Company as an anti-heart failure drug. It was first approved by the FDA in the United States in 1987, officially launched in the United States in 1992, and then successively launched and sold in the United Kingdom, France, Germany, the Netherlands, Belgium and other countries.

[0005] Milrinone is a phosphodiesterase inhibitor, a derivative of amirinone, and has the same mechanism of action as amirinone. It is effective for oral and intravenous administration, and has both positive inotropic effect and vasodilating effect. It is suitable for short-term treatment of patients with severe congestive heart failure who are ineffective for conventional maintenance treatment, and has 10-30 times stronger efficacy than amirinone, better tolerance, and fewer adverse reactions. Its positive inotropic effect is mainly through the inhibition of phosphodiesterase, increasing the concentration of cyclic adenosine monophosphate (CAMP) in myocardial cells, increasing intracellular calcium, and strengthening myocardial contractility and increasing cardiac output. It is generally considered to be a high-efficiency, low-toxicity, non-digitalis, non-pseudo-steroidal inotropic drug. It has a significant effect on severe heart failure, pulmonary edema caused by ischemic heart disease and dilated cardiomyopathy, and is superior to dopamine. It has fewer adverse reactions and does not increase heart rate. Therefore, the drug plays an increasingly important role in the treatment of congestive heart failure (CHF) and peripheral vasodilation.

[0006] In reality, due to poor water solubility of milrinone, and heavy adverse reactions when taken orally, although the prior art discloses some methods for improving the solubility or stability of milrinone, no ideal effect is obtained, and the poor absorption of milrinone is not improved. For example, patent CN1951919A discloses a series of inorganic acid salts of milrinone for preparing injection lyophilized preparations, which can improve the solubility of milrinone, but the stability problem still exists; for example, patent CN102558044A discloses a crystallization method of milrinone, the obtained milrinone has high purity and good crystal form, but the physicochemical properties of milrinone are not improved; in addition, patent CN106361710A describes that, in order to solve the problems of poor stability of lactic acid milrinone in the prior art, easy degradation and significant increase of related substances, a certain amount of vitamin E and glutathione are added in the prescription, and a new crystal form is used to increase the stability of the injection and reduce the generation of degradation reaction, but the use of the new crystal form does not overcome the problem of poor solubility of milrinone.

[0007] Therefore, it is an urgent problem for those skilled in the art to provide a crystal form of milrinone with good solubility, high stability and good drug prospects. SUMMARY

[0008] In view of the poor solubility and low stability of milrinone provided by the prior art, the present application aims to provide a new crystal form of milrinone with high solubility and stability, i.e. milrinone-cinnamic acid crystal. In addition, the present application provides a method for preparing the milrinone-cinnamic acid co-crystal, which is simple, convenient and suitable for industrial production.

[0009] The specific technical content of the present application is as follows:

[0010] In one aspect, the present application provides a high-purity milrinone-cinnamic acid crystal.

[0011] Preferably, the milrinone-cinnamic acid crystal has characteristic peaks at 6.5±0.2°, 10.4±0.2°, 15.6±0.2°, 17.2±0.2°, 21.3±0.2°, 23.8±0.2°, 28.3±0.2° in the X-ray diffraction spectrum expressed in 2θ using Cu-Kα radiation.

[0012] Preferably, the mirinone-cinnamic acid crystal has an X-ray diffraction spectrum using Cu-Ka radiation with characteristic peaks at 6.5±0.2°, 10.4±0.2°, 13.0±0.2°, 13.8±0.2°, 15.6±0.2°, 17.2±0.2°, 21.3±0.2°, 23.8±0.2°, 24.9±0.2°, 25.5±0.2°, 26.6±0.2°, 26.9±0.2°, 28.3±0.2°, 30.7±0.2°, 31.5±0.2°, 40.9±0.2°.

[0013] Preferably, the mirinone-cinnamic acid crystal has an X-ray powder diffraction spectrum as shown in the figure. Figure 1

[0014] Preferably, the mirinone-cinnamic acid crystal has a molecular formula of C 12 H 12 N4O4S, and crystallographic parameters are: triclinic system, space group P-1, cell parameters are: a = 7.0542(2), b = 8.0707(2), c = 17.2349(3), α = 77.502(2)°, β = 85.946(2)°, γ = 71.680(2)°, and cell volume V = 909.41(4).

[0015] In another aspect, the present application provides a method for preparing the mirinone-cinnamic acid crystal, comprising the following steps:

[0016] dissolving mirinone and cinnamic acid in a mixed solvent, heating and stirring, filtering, cooling and crystallizing, filtering, and drying to obtain the mirinone-cinnamic acid crystal.

[0017] Preferably, the solvent is selected from a mixed solvent of methanol and ethanol, isopropanol, acetonitrile, acetone, and water, and particularly preferably a mixed solvent of methanol and acetonitrile.

[0018] Preferably, the mass-volume ratio of mirinone to the mixed solvent is 5-9:1, and preferably 7-8:1, wherein the mass is in mg and the volume is in mL.

[0019] Preferably, the molar ratio of mirinone to cinnamic acid is 1:0.9-1.1, and preferably 1:1.0.

[0020] Preferably, the heating temperature is 50-70°C, and preferably 58°C.

[0021] Preferably, the cooling and crystallizing temperature is 5-15°C, and further preferably 8-12°C.

[0022] Preferably, the crystallization time is 48-96h.​

[0023] Preferably, the drying temperature is 45-65℃, and the drying time is 8-12h.

[0024] In still another aspect, the present application provides a pharmaceutical composition containing the milrinone-cinnamic acid crystal and other pharmaceutically acceptable components.

[0025] Preferably, the other pharmaceutically acceptable components can be co-used pharmaceutically active ingredients and / or pharmaceutically acceptable auxiliary ingredients.

[0026] Compared with the prior art, the present application has the following technical effects:

[0027] The milrinone-cinnamic acid crystal provided by the present application has the advantages of simple preparation method, easy control of crystallization process, and good reproducibility. The milrinone-cinnamic acid crystal has significantly enhanced stability and solubility compared with free base and its crystal form, thereby being beneficial to the storage, transportation and application in preparation of the product, and having enhanced bioavailability and absorption performance. The pharmaceutical polycrystal provided by the present application has better physicochemical properties, which provides a better drug raw material for the treatment of diseases in the clinic, and has great clinical research and development value. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 : X-ray powder diffraction pattern of milrinone-cinnamic acid.

[0029] Figure 2 : ORTEP diagram of milrinone-cinnamic acid.

[0030] Figure 3 : Hydrogen bond diagram of milrinone-cinnamic acid. DETAILED DESCRIPTION

[0031] The present application will be further described below by examples, and it should be understood that the examples of the present application are only used to illustrate the present application, but not to limit the present application, so that simple improvements of the present application under the premise of the method of the present application are within the scope of the present application.

[0032] Example 1

[0033] Milrinone (84.6 mg) and cinnamic acid (59.5 mg) were dissolved in a mixed solvent of methanol (5.5 mL) and acetonitrile (5.5 mL), and heated and stirred in a 58℃ water bath until completely dissolved. After slowly cooling to 8-12℃, temperature control was performed and crystallization was carried out for 72h. Filtration was performed, and vacuum drying was performed at 60℃ for 10h to obtain the milrinone-cinnamic acid crystal, with a yield of 98.73% and a purity of 99.96%.

[0034] Example 2

[0035] Dissolve milrinone (63.63 mg) and cinnamic acid (40.0 mg) in a mixed solvent of methanol (5.0 mL) and ethanol (5.0 mL), heat and stir in a water bath at 50°C until completely dissolved, slowly cool to 8-12°C, control the temperature and stand for crystallization for 48 h, filter, and vacuum dry at 60°C for 12 h to obtain milrinone-cinnamic acid crystals, yield: 97.7%, purity: 99.94%.

[0036] Example 3

[0037] Dissolve milrinone (46.0 mg) and cinnamic acid (21.5 mg) in a mixed solvent of methanol (5 mL) and acetone (5 mL), heat and stir in a water bath at 58°C until completely dissolved, slowly cool to 8-12°C, control the temperature and stand for crystallization for 72 h, filter, and vacuum dry at 60°C for 10 h to obtain milrinone-cinnamic acid crystals, yield: 94.3%, purity: 99.95%.

[0038] Example 4

[0039] Dissolve milrinone (84.6 mg) and cinnamic acid (59.5 mg) in a mixed solvent of methanol (5.5 mL) and acetonitrile (5.5 mL), heat and stir in a water bath at 80°C until completely dissolved, slowly cool to 20°C, control the temperature and stand for crystallization for 36 h, filter, and vacuum dry at 60°C for 10 h to obtain milrinone-cinnamic acid crystals, yield: 95.39%, purity: 99.89%.

[0040] Confirmation of crystal structure

[0041] The X-ray crystal data of the pharmaceutical crystal of milrinone described in the present application were collected on a Japan Rigaku XtaLAB Synergy model instrument, the test temperature was 293(2) K, Cu-Ka radiation was used, the data were collected in an omega scan mode and Lp correction was performed. The structure was solved by direct method, all non-hydrogen atoms were found by difference Fourier method, and all hydrogen atoms on carbon and nitrogen were obtained by theoretical hydrogenation. The structure was refined by least squares method.

[0042] The crystallographic data of the milrinone-cinnamic acid crystal form prepared in the present application (as shown in Table 1) are as follows: triclinic crystal system, space group P-1, cell parameters: a = 7.0542(2), b = 8.0707(2), c = 17.2349(3), α = 77.502(2)°, β = 85.946(2)°, γ = 71.680(2)°, and cell volume V = 909.41(4).

[0043] Table 1 Main crystallographic data of milrinone-cinnamic acid crystals

[0044]

[0045]

[0046] The ORTEP chart of the milrinone-cinnamic acid crystal of the present application shows that one molecule of milrinone and one molecule of cinnamic acid are contained in the crystalline form, as shown in the attached Figure 1. The hydrogen bond chart of the milrinone-cinnamic acid crystal of the present application is shown in the attached Figure 2. According to the above crystallographic data, the characteristic peaks in the X-ray powder diffraction chart (Cu-Ka) thereof are shown in the attached Figure 3 and Table 2. Figure 2 Figure 3 Figure 1

[0047] Table 2 PXRD peaks of the milrinone-cinnamic acid crystal

[0048]

[0049] Comparative Example 1

[0050] In a 1000 mL three-necked flask, 4-methylpyridine 93.0 g, chloroform 500 mL were added, and the temperature was controlled below 50°C by placing the flask in an ice water bath. Acetyl chloride 80.0 g was added dropwise, and after the addition was completed, the temperature was raised to 55°C, and the reaction was carried out for 2.5 h. After the reaction was completed, the system was cooled in an ice bath, and saturated sodium carbonate aqueous solution was added dropwise to adjust the pH to 5-7. Then, 30.0 g of sodium hydroxide solution (30 wt%) was added, and the reaction was carried out at 30-50°C for 2.5 h. After the reaction was completed, the layers were separated, and the water layer was removed. After drying with anhydrous sodium sulfate and recovering the solvent, the fraction at 100-105°C / 217 kPa was collected by distillation, to obtain 1-(4-pyridyl)-2-propanone.

[0051] In a 500 mL round-bottom flask, 1-(4-pyridyl)-2-propanone 60.0 g was added, and 40.5 g of triethyl orthoformate, 92.2 g of acetic anhydride, and 80.0 g of glacial acetic acid were added to the flask with stirring. The reaction was carried out at 35-45°C for 4 h, and after the raw material was consumed, the solvent was removed by concentration under reduced pressure at 80°C, to obtain a dark red oil. The oil was used directly in the next step without purification.

[0052] In a 5000 mL flask, 600 mL of anhydrous methanol and the above oil were added, and 64.0 g of a-cyanacetamide and 210 g of 50% sodium hydroxide solution were added with stirring. The reaction was carried out for 1.5 h. After the reaction was completed, the pH was adjusted to 6.5-7.2 with acetic acid solution, and a solid was precipitated. The solid was filtered to obtain crude milrinone. The solid was recrystallized from an ethanol-water system, to obtain white milrinone crystals.

[0053] Comparative Example 2

[0054] ​​​The 10 g of milbemycin was placed in a 500 mL beaker, and 0.1 N aqueous sodium hydroxide solution was added dropwise while stirring to dissolve it, and the pH was adjusted to 7-8. A solution having an amount 5 times the volume of the solution was added, and the mixture was cooled to precipitate a white solid. The solid was filtered, the filter cake was washed twice with acetone, and the cake was dried at 105°C for 2 hours to obtain milbemycin sodium salt.

[0055] Comparative Example 3

[0056] The 10 g of milbemycin was placed in a 500 mL beaker, and 0.1 N aqueous hydrochloric acid solution was added dropwise while stirring to dissolve it, and the pH was adjusted to 4-4.5. A solution having an amount 5 times the volume of the solution was added, and the mixture was cooled to precipitate a white solid. The solid was filtered, the filter cake was washed twice with acetone, and the cake was dried at 105°C for 2 hours to obtain milbemycin hydrochloride.

[0057] Comparative Example 4

[0058] Milbemycin (169.0 mg) and gallic acid (136.1 mg) were mixed in equimolar ratio, and 50 μL of water was added gradually and the slurry was ground thoroughly for 45 minutes. The ground powder sample was dissolved in a minimum amount of a mixed solvent of methanol / acetonitrile / H2O (v 甲醇 : v 乙腈 : v 水 = 2:1:1) at 60°C with rapid and vigorous stirring for about 3 hours. After cooling to room temperature, the resulting reaction mixture was filtered. To the filtrate, the above powder sample was added as a seed crystal, and the solution was left to evaporate slowly for 1 day to obtain colorless crystals.

[0059] Stability Test

[0060] The specific stability test method was performed according to the guidance method for stability test in the fourth part of Chinese Pharmacopoeia.

[0061] High temperature test: The test sample was placed in an appropriate clean container, and was left at 60°C for 10 days. Samples were taken at the 5th day and the 10th day, and the purity was measured by HPLC.

[0062] High humidity test: The test sample was placed in a constant humidity airtight container, and was left at 25°C for 10 days under the condition of relative humidity 90%±5%. Samples were taken at the 5th day and the 10th day, and the purity was measured by HPLC.

[0063] Strong light irradiation test: The test sample was placed in a light irradiation device equipped with a daylight lamp, and was left for 10 days under the condition of irradiance 4500 lx±500 lx. Samples were taken at the 5th day and the 10th day, and the purity was measured by HPLC.

[0064] Table 3 Stability test results of milbemycin-cinnamic acid crystals

[0065]

[0066]

[0067] The experimental results show that the milrinone-cinnamic acid crystal prepared in the embodiment has high purity, and the sample purity changes little under high temperature, high humidity and strong light conditions, and the stability is good.

[0068] Solubility test

[0069] Method: 10ml of medium (water, 0.01mol / L HCl solution) was respectively taken in a shirn bottle, and an excess of the sample to be tested was added, the shirn bottle was sealed and placed in a 25℃ constant temperature water bath for stirring for 1 hour, filtered through a filter membrane, and the filtrate was taken; the absorbance was respectively measured at a wavelength of 270nm, and the solubility was calculated by testing the absorbance of the standard control.

[0070] Table 4 Solubility test results of the milrinone-cinnamic acid crystal

[0071]

[0072] The experimental results show that the milrinone-cinnamic acid crystal provided by the application has significantly improved solubility in 0.01mol / L HCl and water compared with other crystal forms of milrinone, which is conducive to its application in oral preparations.

[0073] Pharmacokinetic study

[0074] Method: The in vivo PK test was carried out by a single dose oral administration method, male SD rats (220-260g) were fed in a constant humidity of 0%-60% in a quiet environment, the temperature was 25±1℃, and the light was rhythmic from 7am to 7pm. The PK test was strictly carried out according to the Laboratory Management Guide issued by the Ministry of Science and Technology of China. Before the experiment, the test rats were randomly divided into three groups (n=5 each), and they were allowed to drink water freely and fast overnight. All the tested samples were suspended in vegetable oil, and then a single dose of 10mg / kg milrinone or its equivalent was orally administered. After administration, 0.5mL of blood sample was collected at the designed time point according to the administration, and the blood drug concentration of milrinone was detected according to the literature method.

[0075] Table 5 Pharmacokinetic test results of the milrinone-cinnamic acid crystal

[0076]

[0077] The test results show that the milrinone-cinnamic acid crystal form provided by the application reaches a higher peak concentration at a faster speed compared to other crystal forms of milrinone, which is the same as the solubility tendency, and provides conditions for milrinone and a large amount of drugs to be rapidly absorbed into the blood. Compared to other crystal forms of milrinone, the half-life of the milrinone-cinnamic acid crystal form is prolonged, so that the drug stays in the body for a longer time, thereby providing the possibility of obtaining a long-term therapeutic effect.

Claims

1. A mirin-cinnamic acid crystal, characterized by, X-ray diffraction spectrum using Cu-Kα radiation has characteristic peaks at at least 6.5±0.2°, 10.4±0.2°, 15.6±0.2°, 17.2±0.2°, 21.3±0.2°, 23.8±0.2°, 28.3±0.2° in terms of 2θ; the milrinone-cinnamic acid crystal has a molecular formula of C 12 H 12 N4O4S, and the crystallographic parameters are: triclinic system, space group P-1, cell parameters are: a=7.0542(2), b=8.0707(2), c=17.2349(3), α=77.502(2)°, β=85.946(2)°, γ=71.680(2)°, and cell volume V=909.41(4).

2. The mirinorcinnamic acid crystal of claim 1, wherein X-ray diffraction spectrum using Cu-Kα radiation has characteristic peaks at at least 6.5±0.2°, 10.4±0.2°, 13.0±0.2°, 13.8±0.2°, 15.6±0.2°, 17.2±0.2°, 21.3±0.2°, 23.8±0.2°, 24.9±0.2°, 25.5±0.2°, 26.6±0.2°, 26.9±0.2°, 28.3±0.2°, 30.7±0.2°, 31.5±0.2°, 40.9±0.2° in terms of 2θ.

3. The mirinorcinnamic acid crystal of claim 1, wherein X-ray powder diffraction spectrum using Cu-Kα radiation has characteristic peaks as shown in Figure 1.

4. A process for preparing mirinocine-cinnamic acid crystals as claimed in claim 1, characterized by, The specific preparation steps include: dissolving milrinone and cinnamic acid in a mixed solvent, heating and stirring, filtering, cooling and crystallizing, filtering, and drying to obtain milrinone-cinnamic acid crystals.

5. The preparation method of the mirin-cinnamic acid crystal according to claim 4, characterized by, The mixed solvent is selected from the group consisting of methanol and ethanol, isopropanol, acetonitrile, acetone, and water.

6. The preparation method of the mirin-cinnamic acid crystal according to claim 4, wherein The mixed solvent is a mixture of methanol and acetonitrile.

7. The preparation method of the mirin-cinnamic acid crystal according to claim 4, wherein The mass-volume ratio of milrinone to the mixed solvent is 5-9:1, wherein the mass is in mg and the volume is in mL.

8. The preparation method of the mirin-cinnamic acid crystal according to claim 4, wherein The mass-volume ratio of milrinone to the mixed solvent is 7-8:1, wherein the mass is in mg and the volume is in mL.

9. The preparation method of the mirin-cinnamic acid crystal according to claim 4, wherein The molar ratio of milrinone to cinnamic acid is 1:0.9-1.

1.

10. The method of preparing mirinor-cinnamic acid crystals according to claim 4, wherein The molar ratio of milrinone to cinnamic acid is 1:1.

0.

11. The preparation method of the mirin-cinnamic acid crystal according to claim 4, wherein The cooling and crystallization temperature is 5-15℃.

12. The preparation method of the mirin-cinnamic acid crystal according to claim 4, wherein The cooling and crystallization temperature is 8-12℃.

13. The preparation method of the mirin-cinnamic acid crystal according to claim 4, wherein The drying temperature is 55-65℃, and the drying time is 8-10h.

14. A pharmaceutical composition, characterized by, The composition contains the milrinone-cinnamic acid crystals of any one of claims 1-3.

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