COMPOSITION CONTAINING LEGOAMODIPINE BESYLATE HYDRATE AND PREPARATION METHOD THEREFOR

Abstract

The present invention relates to a composition comprising levamlodipine besylate hydrate and its production, pharmaceutical preparations and use, especially the composition of (S)-2-[(2-aminoethoxy)methyl]-4-(2-chlorophenyl)-6-methyl-1,4-dihydro-3,5-pyridinedicarboxylic acid-3-ethyl ester, 5-methyl ester benzenesulfonic acid hydrate and its production method and use. The composition of levamlodipine besylate crystallized in pure water and dried is easy for industrial production, has no organic solvent residue, good thermal stability and good dissolution amount in solid-form preparations.

Claims

1-60. (canceled)

61. A composition comprising levamlodipine besylate hydrate having different crystalline water content, wherein the levamlodipine besylate hydrate has a molecular formula of C.sub.20H.sub.25ClN.sub.2O.sub.5C.sub.6H.sub.6O.sub.3S.Math.nH.sub.2O, wherein 1<n<2, and has the following structural formula: ##STR00002## and wherein the composition comprises 4.5%-5.8% of crystalline water.

62. The composition of claim 1, wherein the composition comprises 4.5%-5.5% of crystalline water.

63. The composition of claim 1, wherein the composition comprises 4.5%-5.4% of crystalline water.

64. The composition of claim 1, wherein the composition is prepared by crystallizing and precipitating the levamlodipine besylate hydrate in pure water containing no organic solvent, separating, and drying.

65. A drug formulation comprising the composition of claim 1 and a pharmaceutically acceptable carrier.

66. The drug formulation of claim 65, being a combination preparation and further comprising a pharmacologically acceptable pharmaceutical ingredient.

67. A fast-sustained-release preparation of levamlodipine besylate comprising the composition of claim 61, wherein the preparation is formulated in the following mass ratios: a sustained-release layer comprising: levamlodipine besylate calculated in levamlodipine 4-5 polyvinylpyrrolidone 30-40 hydroxypropyl cellulose 55-65 microcrystalline cellulose 17-30 magnesium stearate 0.1-1; and a fast-release layer comprising: levamlodipine besylate calculated in levamlodipine 1-2 microcrystalline cellulose 40-45 pregelatinized starch 35-45 cross-linked polyvinylpyrrolidone 1-8 polyvinylpyrrolidone 5-15 magnesium stearate 0.1-1.

68. A fast-sustained-release preparation of levamlodipine besylate comprising the composition of claim 62, wherein the preparation is formulated in the following mass ratios: a sustained-release layer comprising: levamlodipine besylate calculated in levamlodipine 4-5 polyvinylpyrrolidone 30-40 hydroxypropyl cellulose 55-65 microcrystalline cellulose 17-30 magnesium stearate 0.1-1; and a fast-release layer comprising: levamlodipine besylate calculated in levamlodipine 1-2 microcrystalline cellulose 40-45 pregelatinized starch 35-45 cross-linked polyvinylpyrrolidone 1-8 polyvinylpyrrolidone 5-15 magnesium stearate 0.1-1.

69. A fast-sustained-release preparation of levamlodipine besylate comprising the composition of claim 63, wherein the preparation is formulated in the following mass ratios: a sustained-release layer comprising: levamlodipine besylate calculated in levamlodipine 4-5 polyvinylpyrrolidone 30-40 hydroxypropyl cellulose 55-65 microcrystalline cellulose 17-30 magnesium stearate 0.1-1; and a fast-release layer comprising: levamlodipine besylate calculated in levamlodipine 1-2 microcrystalline cellulose 40-45 pregelatinized starch 35-45 cross-linked polyvinylpyrrolidone 1-8 polyvinylpyrrolidone 5-15 magnesium stearate 0.1-1.

70. A fast-sustained-release combination tablet comprising levamlodipine besylate and folic acid, being formulated in the following mass ratios: a sustained-release combination layer comprising: levamlodipine besylate calculated in levamlodipine 4-5 polyvinylpyrrolidone 30-40 hydroxypropyl cellulose 55-65 microcrystalline cellulose 17-30 magnesium stearate 0.1-1; and a fast-release combination layer comprising: levamlodipine besylate calculated in levamlodipine 1-2 folic acid 0.1-1 microcrystalline cellulose 40-45 pregelatinized starch 35-45 cross-linked polyvinylpyrrolidone 1-8 polyvinylpyrrolidone 5-15 magnesium stearate 0.1-1, wherein levamlodipine besylate is included as a composition of levamlodipine besylate hydrate comprising levamlodipine besylate hydrate having different crystalline water content, and wherein the levamlodipine besylate hydrate has a molecular formula of C.sub.20H.sub.25ClN.sub.2O.sub.5C.sub.6H.sub.6O.sub.3S nH.sub.2O, wherein 1<n<2.

71. The fast-sustained-release combination tablet of claim 70, being formulated in the following mass ratios: the sustained-release combination layer comprising: levamlodipine besylate calculated in levamlodipine 4.5 polyvinylpyrrolidone 35 hydroxypropyl cellulose 60 microcrystalline cellulose 23.9 magnesium stearate 0.5; and the fast-release combination layer comprising: levamlodipine besylate calculated in levamlodipine 1.5 folic acid 0.4 microcrystalline cellulose 40.5 pregelatinized starch 40 cross-linked polyvinylpyrrolidone 5 polyvinylpyrrolidone 10 magnesium stearate 0.5.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0046] FIG. 1: a photo of the porridge-like crystals of levamlodipine besylate hydrate in water

[0047] FIG. 2: powder diffraction patterns of levamlodipine besylate hydrate compositions, in the figure from top to bottom are the reference standard's simulated powder diffraction pattern of levamlodipine besylate with 1.5 crystalline water molecules (crystallization solvent actone+water), the powder diffraction pattern of a levamlodipine besylate hydrate composition produced according to the present invention, the reference standard's simulated powder diffraction pattern of levamlodipine besylate with 2.5 crystalline water molecules (crystallization solvent ethanol+water)

[0048] FIG. 3: a chromatogram of sample 1 (crystalline water content 2.0%) placed in a 45° C. oven for 0 days

[0049] FIG. 4: a chromatogram of sample 1 (crystalline water content 2.0%) placed in a 45° C. oven for 5 days

[0050] FIG. 5: a chromatogram of sample 2 (crystalline water content 4.5%) placed in a 45° C. oven for 0 days

[0051] FIG. 6: a chromatogram of sample 2 (crystalline water content 4.5%) placed in a 45° C. oven for 5 days

[0052] FIG. 7: a chromatogram of sample 3 (crystalline water content 5.4%) placed in a 45° C. oven for 0 days

[0053] FIG. 8: a chromatogram of sample 3 (5.4% water of crystallization) placed in a 45° C. oven for 5 days

[0054] FIG. 9: the dissolution amount of tablets of the levamlodipine besylate hydrate compositions correlate with the crystalline water content of the hydrate.

[0055] FIG. 10: a mean plasma concentration-time curve of beagles after oral administration of 10 mg of a test preparation of a levamlodipine besylate hydrate composition and a reference preparation.

SPECIFIC MODES FOR CARRYING OUT THE INVENTION

Example 1 Preparation and Physical and Chemical Parameter Test of Levamlodipine Besylate Hydrate Composition Satisfying Requirements of Solid-Form Preparation Plain Tablets

[0056] Preparation of Levamlodipine Besylate Hydrate Composition

[0057] Weigh 51 kilograms of purified water, put the purified water into a reactor through a feeding port and stir, heat up to 46° C., add 1.60 kilograms of levamlodipine, and an aqueous solution with 0.65 kilograms of benzenesulfonic acid dissolved in 0.2 kilograms of water. The molar ratio of levamlodipine to benzenesulfonic acid is about 1:1.05 to ensure that all levamlodipine reacts with benzenesulfonic acid to generate organic salts. The total amount of purified water is 51.2 kilograms, which is 32 times the feeding amount of levamlodipine. After completely dissolving, it is transferred to a crystallization bucket, placed in a freezer for crystallization, and the crystals are porridge-like in water, see FIG. 1, temperature controlled to be within the range of-5° C. to 10° C., the temperature in this experiment is controlled at −5° C., the time is ≥12 hours, and then centrifuged at a speed of 2000 r/min, and filtered. Then rinse with 2 kg of purified water: after stopping the centrifuge, pour the purified water on the surface of the filter bag, then start the centrifuge machine to centrifuge, and then dry. Room temperature: 20° C., humidity: 52%, air flow through the through-air oven: 12000 m.sup.3/hour, drying temperature 30±2° C., drying time 1.5 hours, crystalline water content 5.3%.

[0058] Test of Physical and Chemical Parameters of the Levamlodipine Besylate Hydrate Composition

[0059] Single crystals of levamlodipine besylate acetone hydrate and levamlodipine besylate ethanolate hydrate containing 1.5 crystalline wate molecules and 2.5 crystalline water molecules and containing organic solvents can be prepared in acetone+water and ethanol+water, respectively. However, the single crystals of levamlodipine besylate hydrate cannot be crystallized in pure water yet, and only a composition of levamlodipine besylate hydrate can be produced. The data of the powder of the composition prepared by the method of the present invention is similar to the simulated powder data of the single crystals of the above-mentioned levamlodipine besylate with 1.5 crystalline water molecules and 2.5 crystalline water molecules. The test conditions are as follows:

[0060] The X-ray powder diffractometer model D8 Advance used is an apparatus from Bruker, Germany. The samples are, respectively: the composition of levamlodipine besylate hydrate prepared in Example 1, single crystals of levamlodipine besylate with 1.5 crystalline water molecules, and single crystals of levamlodipine besylate with 2.5 crystalline water molecules.

[0061] Test method of the crystal form of levamlodipine besylate powder: place the powder sample on the sample plate and flatten it, and use the D8 Advance instrument to collect the diffraction data. Instrument setup parameters: Cu target source (Kα), wavelength λ=1.5406 Å. The test voltage and current are 40 kV and 40 mA respectively, the test range is 5-45°, and the scanning speed is 0.02°/s. The powder diffraction data collected are directly compared to powder data simulated from the X-ray single crystal data of the reference standards containing 2.5 and 1.5 water molecules. The simulated powder data of the reference standards are based on X-single crystal data, and are obtained using Mercury software. The diffraction results are shown in FIG. 2. It can be seen that the crystalline water content of the levamlodipine besylate hydrate composition powder is between levamlodipine besylate with 1.5 crystalline water molecules and levamlodipine besylate with 2.5 crystalline water molecules.

Example 2 Production of Levamlodipine Besylate Hydrate Composition with Specific Crystalline Water Content

[0062] The unseparated levamlodipine besylate hydrate prepared in pure water should be a compound with a fixed proportion of crystalline water, which is difficult to industrially produce on a large scale. Because one of the crystalline water molecules has a low dissociation temperature, that crystal water molecule is very easy to dissociate into the air. To keep the crystalline water content of the hydrate unchanged, the compound needs to be strictly kept at a relatively low temperature, under corresponding humidity and atmospheric pressure. Obviously, such prepared levamlodipine besylate hydrate is not pharmaceutically needed and is not suitable for the requirements of direct compression. However, the preparation of a levamlodipine besylate hydrate composition can meet the requirement % of various types of pharmaceutical preparation %.

[0063] The stability of the levamlodipine besylate hydrate composition is related to temperature. Obviously, the lower the temperature, the more stable the compound. Levamlodipine besylate hydrate compositions with corresponding crystalline water content can be obtained under the drying conditions of Example 1, and meet the requirements of relevant quality standards.

TABLE-US-00001 TABLE 1 Drying Time and Water Content Drying Time (mins) H.sub.2O% 0 20.9 5 18.9 10 16.1 15 14.2 20 12.1 25 9.3 30 8.5 35 8.2 40 7.9 45 7 5 50 7.1 55 6.9 60 6.7 65 6.6 70 6.4 75 6.2 80 5.8 85 5.6 90 5.3 95 5.1 100 4.8 105 4.6 110 4.4 115 4.2

[0064] To obtain a levamlodipine besylate composition with lower crystalline water content requires lower air humidity to reduce drying time. The lower the crystalline water content of the levamlodipine besylate composition, the more voids the dissociated crystalline water will leave on the particle surface of the levamlodipine besylate composition, the larger the contact area between levamlodipine besylate and oxygen in the air, the more opportunities for oxidation reaction to occur, which is not conductive to long-term storage of pharmaceutical preparations.

Example 3 Material Properties of Levamlodipine Besylate Hydrate Composition Granules

[0065] In order to meet the requirements for the production of solid-form preparation plain tablets, the preparation is a direct compression method. As such, the granules of the active pharmaceutical ingredient need to pass through a sieve below 10 mesh to ensure content uniformity. The granules of the levamlodipine besylate hydrate composition needs to be finished, and the particle size for the finishing of granulation is controlled to be 10 mesh. The levamlodipine besylate hydrate composition with a relatively high crystalline water content is soft, and they easily stick together during the finishing of granulation process and cannot meet the requirements of the finishing of granulation. A hydrate composition with a crystalline water content of less than 5.8% can meet the requirements of the finishing of granulation.

[0066] The composition of levamlodipine besylate hydrate with a crystalline water content of 3-6% prepared in Example 1 is dried to obtain compositions of levamlodipine besylate hydrate with different crystalline water content. By testing its physical properties, it is found that the granules of the samples with a crystalline water content of more than 5.8% become soft, and the active pharmaceutical ingredient and auxiliary materials easily stick together, which is not suitable for making pharmaceutical preparations. The crystalline water content of the hydrate being controlled below 5.5% can better meet the requirements of making pharmaceutical preparations.

[0067] 1. Production of Samples: the method of Example 2 is used to prepare samples with corresponding water content.

[0068] 2. Results:

TABLE-US-00002 TABLE 2 Granulation Results Sample Number of Crystalline Crystalline Water Water Sieving Content % Molecules Stickiness Performance 6.0% 2.01 stickiness The materials between which pass particles through the sieve stick together even when slightly squeezed 5.9% 1.98 stickiness The materials between which pass particles through the sieve are less sticky when slightly squeezed 5.8% 1.94 essentially no The materials stickiness which pass between through the sieve particles have essentially no stickiness 5.5% 1.83 no stickiness The materials between which pass particles through the sieve have no stickiness

Example 4 Thermal Stability of Levamlodipine Besylate Hydrate Compositions

[0069] Using the method of Example 2 of the present invention, compositions with different crystalline water content can be prepared and used as samples to test thermal stability.

[0070] The samples are placed in amber glass bottles, placed in an oven, and the temperature is adjusted to 45° C., and the relevant substances are tested after settled for 5 days.

[0071] The test method is as follows: The chromatographic conditions and system applicability test: Use octadecylsilane-bonded silica gel as the filler; methanol-0.03 mol/L potassium dihydrogen phosphate solution (75:25) as the mobile phase, the detection wavelength is 238 nm, the number of theoretical plates should be no less than 500 calculated in levamlodipine.

[0072] Preparation of the Test Solution: Take about 17 mg of this product, precisely weigh it, put it in a 100 ml volumetric flask, add a mobile phase to dissolve and dilute to the mark, shake well, precisely measure 5 ml, put it in a 25 ml volumetric flask, add a mobile phase to the mark, shake well to obtain the solution.

[0073] Determination Method: Precisely measure 10 μl of the reference solution and the test solution, inject into the liquid chromatography machine, record the chromatogram, measure the peak area, and calculate by the external standard method.

[0074] The test data is as follows:

TABLE-US-00003 TABLE 3 impurities Before Heating Sample Placed in 45° C. of Sample Oven for 5 Days Crystalline Retention Retention Sample Content Time of Impurity Time of Impurity Name Water Impurities Content Impurities Content Sample 1 2.0% — None 4.304 min 0.07% Sample 2 4.5% — None — None Sample 3 5.4% — None — None

[0075] Chromatograms of samples 1-3 after being placed in a 45° C. oven for 5 days are shown in FIGS. 3 to 8.

[0076] It can be seen that the levamlodipine besylate hydrate prepared in pure water is filtered and dried, and in the obtained product, the thermal stability of the levamlodipine besylate hydrate composition with a crystalline water content of 4.5-5.4% is better than other compositions with a different crystal water content. There is no obvious change in the appearance of sample 1. After the partial loss of the crystalline water, no fusion occurs between the particles, and the voids left inside the particles are prone to oxidation. For samples 2 and 3, less crystalline water is lost, less voids are left inside the particles, and the degree of oxidation is reduced.

[0077] The precipitated crystals obtained from the preparation of levamlodipine besylate in pure water is a hydrate, levamlodipine besylate hydrate of levamlodipine besylate and 2 crystalline water molecules. However, as shown in Example 3 above, for levamlodipine besylate hydrate with 2 crystalline water molecules, the active pharmaceutical ingredient and auxiliary materials easily stick together, which is not suitable for making pharmaceutical preparations. And for the levamlodipine besylate hydrate prepared by using an organic solvent and water, organic solvents such as alcohols, ketones, etc. will react with levamlodipine besylate to produce benzenesulfonate ester products in the preparation process. It is common knowledge in the art that benzenesulfonate esters are genotoxic (e.g., see Shao Xiaowei, Research on Genotoxicity Detection and Synthesis of Benzenesulfonate Esters, Jilin University Master Thesis, CNKI China Knowledge Network Master Thesis Database, published on May 1, 2019). In addition, in the prepared levamlodipine besylate hydrate product, organic solvent components will inevitably remain, and these organic components are harmful to various degrees. For example, it is common knowledge in the art that acetone has toxic effects to the nervous system and mucous membranes. To sum up, it can be seen that the hydrate prepared by using organic solvents contains organic molecules bonded thereto. Even though it can have improved stability by adjusting its crystalline water content, compared with the hydrate composition prepared by using pure water in the present invention, it does not have any pharmaceutical advantages due to its toxicity.

Example 5 Dissolution Amount of Solid-Form Preparation of Levamlodipine Besylate Hydrate Composition

[0078] The levamlodipine besylate hydrate composition prepared by the method of Example 2 of the present invention, the hydrate composition has different crystalline water content, and is prepared as tablets according to the following formulation:

TABLE-US-00004 TABLE 4 levamlodipine besylate hydrate  2.5 mg/tablet composition (calculated in levamlodipine) starch 53.75 mg/tablet dextrin 41.25 mg/tablet sodium starch glycolate 25.00 mg/tablet magnesium stearate  1.20 mg/tablet

[0079] Each tablet weighs about 125 mg. Weigh the active pharmaceutical ingredient and the auxiliary materials as required by the number of tablets according to the above formulation and mix them evenly.fwdarw.direct compression, wherein the starch passes through a 60 mesh sieve during granulation.

[0080] Test of Dissolution Amount of Levamlodipine Besylate Hydrate Composition Tablets (each tablet containing 2.5 mg of levamlodipine),

[0081] Test Method for Dissolution Amount: Protect from light. Take this product, follow the test method for dissolution amount (Chinese Pharmacopoeia 2015 Edition Part Four General Principles 0931), use 200 ml of hydrochloric acid solution (9-1000) as the solvent, and operate at 50 revolutions per minute. After 30 minutes, take an appropriate amount of the solution, filter, take the filtrate in the middle of the filtration process as the test solution. In addition, take 17 mg of levamlodipine besylate hydrate reference substance, precisely weigh it, put it in a 100 ml volumetric flask, add 2 ml of methanol to dissolve, add hydrochloric acid solution (9.fwdarw.1000) to dilute to the mark, shake well, precisely measure 5 ml, put it in in a 50 ml volumetric flask, add hydrochloric acid solution (9.fwdarw.1000) to dilute to the mark, shake well, and use it as the reference solution.

[0082] Take the above two solutions, according to the spectrophotometry method (Chinese Pharmacopoeia 2015 Edition Part Four General Principles 0401), measure the respective absorbance at the wavelength of 238 nm, and calculate the dissolution amount of each tablet. The limit is 80% of the label amount, which should comply with the regulations.

[0083] The test results are as follows:

TABLE-US-00005 TABLE 5 Crystalline Crystalline Crystalline Water Dissolution Crystalline Dissolution Water Dissolution Water Dissolution Content Amount Item Water Amount Item Content Amount Item Content Amount (%) (%) No. Content (%) No. (%) (%) No. (%) (%) 1 1.3 89.0 40 3.9 95.9 79 4.1 92.6 118 4.3 92.1 2 1.6 93.3 41 3.9 94.3 80 4.1 98.1 119 4.3 92.7 3 1.7 91.1 42 3.9 90.8 81 4.1 94.4 120 4.3 93.! 4 1.7 92.9 43 3.9 90.0 82 4.1 91.6 121 4.4 88.8 5 1.7 87.8 44 3.9 95.7 83 4.1 89.8 122 4.4 95.3 6 1.7 94.2 45 3.9 95.3 84 4.1 88.7 123 4.4 91.1 7 1.7 94.3 46 3.9 92.8 85 4.1 93.3 124 4.4 97.3 8 1.8 95.0 47 3.9 91.2 86 4.1 94.2 125 4.4 97.7 9 1.8 90.8 48 3.9 96.7 87 4.2 90.3 126 4.4 97.4 10 1.8 87.6 49 3.9 96.7 88 4.2 92.9 127 4.4 91.5 11 1.9 91.2 50 3.9 95.8 89 4.2 92.5 128 4.4 91.6 12 1.9 87.0 51 3.9 9'6.1 90 4.2 97 6 129 4.4 91.8 13 2.0 87.5 52 3.9 93.8 91 4.2 97.9 130 4.4 91.7 14 2.4 99.2 53 3.9 96.5 92 4.2 97.1 131 4.4 94.4 15 2.7 92.0 54 3.9 90.0 93 4.2 93.1 132 4.4 97.9 16 3.0 93.0 55 4.0 93.4 94 4.2 94.1 133 4.5 96.3 17 3.2 96.4 56 4.0 92.0 95 4.2 96.1 134 4.5 92.5 18 3.2 96.4 57 4.0 94.4 96 4.2 89.1 135 4.5 92.3 19 3.3 93.7 58 4.0 93.3 97 4.2 95.5 136 4.5 89.6 20 3.5 90.8 59 4.0 94.5 98 4.2 95.5 137 4.5 96.3 21 3.4 87.7 60 4.0 95.7 99 4.2 95 9 138 4.5 89.4 22 3.4 93.1 61 4.0 94.7 100 4.2 92.4 139 4.5 90.1 23 3.5 90.2 62 4.0 97.7 101 4.2 95.2 140 4.5 92.5 24 3.5 95.2 63 4.0 96.2 102 4.2 97.0 141 4.6 92.9 25 3.5 92.3 64 4.0 90.7 303 4.2 97.9 142 4.6 93.0 26 3.5 95.6 65 4.0 98.1 104 4.3 91.1 143 4.6 95.4 27 3.5 96.0 66 4.0 93.6 105 4.3 92.2 144 4.6 95.3 28 3.5 94.0 67 4.0 88.7 106 4.3 96.1 145 4.6 93.2 29 3.5 93.7 68 4.0 88.1 107 4.3 90.4 146 4.6 96.3 30 3.5 89.1 69 4.0 93.9 108 4.3 97.3 147 4.6 94.3 31 3.5 95.8 70 4.1 93.5 109 4.3 95.6 148 4.6 94.0 32 3.5 96.4 71 4.1 96.5 110 4.3 94.8 149 4.6 96.3 33 3.5 95.6 72 4.1 97.0 311 4.3 91.4 150 4.6 92.9 34 3.6 95.1 73 4.1 96.9 112 4.3 92.8 151 4.6 95.8 35 3.6 92.5 74 4.1 88.9 113 4.3 93.6 152 4.8 92.4 36 3.7 92.9 75 4.1 94.9 114 4.3 94.4 153 4.8 89.8 37 3.7 94.8 76 4.1 95.0 115 4.3 91.0 154 4.9 89.4 38 3.8 89.9 77 4.1 96.9 116 4.3 92.4 155 5.0 90.7 39 3.8 91.3 78 4.1 93.5 117 4.3 95.7 156 5.2 93.7

TABLE-US-00006 TABLE 6 Linear Correlation Coefficients Dependent Variable: Dissolution Amount (%) Independent Variable: Crystalline Water Content (%) Model Summary Degree of Degree of Coefficient Estimates Formula R.sup.2 P Freedom 1 Freedom 2 p-value Constant b1 Linear 0.038 6.144 1 154 0.014 90.776 0.696

[0084] The correlation diagram of the dissolution amount of the levamlodipine besylate hydrate composition tablets and the crystalline water content of the hydrate is shown in FIG. 9.

[0085] The above analysis shows that the dissolution amount is positively correlated to the crystalline water content of the composition. Obviously, the production of a pharmaceutical composition of the levamlodipine besylate hydrate composition can make a solid-form preparation have a better dissolution amount.

Example 6 Levoamlodipine Besylate Hydrate Composition or +Folic Acid Tablet Formulations

[0086]

TABLE-US-00007 TABLE 7 Single-Active-Ingredient and Combination Tablet Formulations Single-Active-Ingredient Formulation Combination Tablet Formulation Materials of Sing- Materials of Active-Ingredient Combination Formulation mg/tablet Formulation mg/tablet calculated in 2.5 calculated in 2.5 levamlodipine 2.5 levamlodipine folic acid 0.4 anhydrous calcium 39.8 anhydrous calcium 39.4 hydrogen phosphate hydrogen phosphate microcrystalline 79 microcrystalline 79 cellulose cellulose sodium starch 2.5 sodium starch 2.5 glycol ate glycolate magnesium stearate 1.2 magnesium 1.2 stearate

[0087] Each tablet weighs about 125 mg. The levamlodipine besylate used in this Example is the levamlodipine besylate hydrate composition prepared by the method of Example 1, but is not limited to the product of Example 1 in actual production and application. The levamlodipine besylate hydrate compositions protected by the present invention may all be used to make corresponding pharmaceutical preparations. According to the above formulation, weigh the active pharmaceutical ingredients and auxiliary materials as required by the number of the tablets and mix them evenly.fwdarw.direct compression.

[0088] Combination tablets can also be compressed into double-layered tablets, that is, two active pharmaceutical ingredients are each mixed with auxiliary materials for double-layered tableting.

Example 7 Levoamlodipine Besylate Hydrate Composition+Bisoprolol Fumarate or +Folic Acid Tablet

[0089]

TABLE-US-00008 TABLE 8 Combination and Triple-Active-Ingredient Tablet Formulations composition of levamlodipine besylate hydrate + composition of levamlodipine besylate hydrate + bisoprolol fumarate bisoprolol fumarate + folic acid tablet Materials of Materials of Triple- Combination Active-Ingredient Formulation mg/tablet Formulation mg/tablet calculated in levamlodipine 2.5 calculated in 2.5 levamlodipine calculated in bisoprolol 2.5 calculated in bisoprolol 2.5 folic acid 0.4 anhydrous calcium 63 anhydrous calcium 63 hydrogen phosphate hydrogen phosphate 0. microcrystalline cellulose 124.65 microcrystalline 124.65 cellulose sodium starch glycolate 4 sodium starch glycolate 4 magnesium stearate 2 magnesium stearate 9

[0090] Each tablet weighs about 200 mg. The levamlodipine besylate used in this Example is the levamlodipine besylate hydrate composition prepared by the method of Example 1, but is not limited to the product of Example 1 in actual production and application. The levamlodipine besylate hydrate compositions protected by the present invention may all be used to prepare corresponding pharmaceutical preparations. According to the above formulation, weigh the active pharmaceutical ingredients and the auxiliary materials as required by the number of tablets and mix them evenly.fwdarw.direct compression.

Example 8 Levoamlodipine Besylate Hydrate Composition+Folic Acid-Fast-Sustained-Release Formulations

[0091]

TABLE-US-00009 TABLE 9 Single-Active-Ingredient or Combination Fast-Sustained-Release Formulations Levamlodipine Besylate Hydrate Composition Levamlodipine Besylate Hydrate Composition + Folic Acid Fast-Sustained-Release Formulation Fast-Sustained Release-Formulation Sustained-Release Sustained-Release Combination Laver (First Layer (First Layer) mg/tablet Layer) mg/tablet calculated in 4.0 calculated in levamlodipine 4.5 levamlodipine polyvinylpyrrolidone 35 polyvinylpyrrolidone 35 hydroxypropyl cellulose 60 hydroxypropyl cellulose 60 microcrystalline cellulose 23.9 microcrystalline cellulose 23.9 magnesium stearate 0.5 magnesium stearate 0.5 calculatedin 1.5 calculated in levamlodipine 1.5 levamlodipine folic acid 0.4 microcrystalline cellulose 40.5 microcrystalline cellulose 40.5 pregelatinized starch 40 pregelatinized starch 40 cross-linked 5 cross-linked 5 polyvinylpyrrolidone polyvinylpyrrolidone polyvinylpyrrolidone 10 polyvinylpyrrolidone 10 magnesium stearate 0.5 magnesium stearate 0.5

[0092] Each bilayer tablet weighs about 220 mg. The levamlodipine besylate used in this Example is the levamlodipine besylate hydrate composition produced by the method of Example 1, but is not limited to the product of Example 1 in actual production and application. The levamlodipine besylate hydrate compositions protected by the present invention may all be used to prepare corresponding pharmaceutical preparations.

[0093] According to the above formulation, weigh the active pharmaceutical ingredients and auxiliary materials as required by the number of tablets and mix them evenly.fwdarw.direct compression. Each tablet contains levamlodipine besylate calculated in levamlodipine 6 mg+folic acid 0.4 mg.

TABLE-US-00010 TABLE 10 Example 9 Levoamlodipine Besylate Hydrate Composition + or Bisoprolol Fumarate or + Folic Add Capsule Formulations Single-Active-Ingredient Combination Capsule Triple-Active-Ingredient Capsule Formulation Formulation Capsule Formulation Materials of Materials of Materials of Single-Active-Ingredient Combination Triple-Active-Ingredient Formulation mg/tablet Formulation mg/tablet Formulation mg/tablet calculated in 2.5 calculated in 2.5 calculated in 2.5 levamlodipine levamlodipine levamlodipine calculated in 2.5 calculated in 2.5 bisoprolol bisoprolol anhydrous 122.5 mg anhydrous 120 mg anhydrous 120 mg calcium calcium calcium hydrogen hydrogen hydrogen phosphate phosphate phosphate

[0094] Each tablet weighs approximately 125 mg. The levamlodipine besylate used in this Example is the levamlodipine besylate hydrate composition prepared by the method of Example 1, but is not limited to the product of Example 1 in actual production and application. The levamlodipine besylate hydrate compositions protected by the present invention may all be used to prepare corresponding pharmaceutical preparations. Weigh the above materials.fwdarw.mix.fwdarw.encapsulate.

Example 10 Production of Oral Liquid Preparation of Levamlodipine Besylate Hydrate Composition

[0095] Solid-form preparations are not suitable for children and people who have difficulty swallowing. An oral liquid of the levamlodipine besylate hydrate composition is suitable for all patients to take. Because levamlodipine besylate hydrate compositions are all suitable for production of oral liquid preparations, the oral liquids prepared by the levamlodipine besylate hydrate compositions do not contain organic solvent residues and do not need adjuvants to be added, 2.5 mg of a levamlodipine besylate hydrate composition is dissolved in 3.5 mg of pure water. Levamlodipine besylate is bitter in taste, and thus a flavoring agent may be added in an appropriate amount, such as aspartatyl chloride phenylalanine methyl ester, which is a dipeptide sweetener, with a sweetness of 180 ˜X° C. 300 times that of sucrose, with good sweetness and high safety, its metabolism does not require insulin, does not cause dental caries, and effectively reduces calorie intake. It is included in USP XXIII edition, and the dosage concentration ranges from 0.1% to 0.6%, suitable for patients with diabetes and obesity.

[0096] For children's medication, an appropriate amount of liquid may be withdrawn with a straw for oral consumption.

[0097] Preparation process, take a levamlodipine besylate hydrate composition equivalent to 10 grams of levamlodipine+flavoring agent (for example, 14 grams of aspartyl chloride phenylalanine methyl ester) dissolved in purified water 14 L.fwdarw.stir.fwdarw.fill 4 ml amber glass bottle-cap the bottle. Storage temperature is cool. The levamlodipine besylate used in this Example is the levamlodipine besylate hydrate composition produced by the method of Example 1, but is not limited to the product of Example 1 in actual production and application. The levamlodipine besylate hydrate compositions protected by the present invention may all be used to produce corresponding pharmaceutical preparations.

Example 11 Acute Toxicity of Levamlodipine Besylate Hydrate Composition Tablets

[0098] In this experiment the tablet formulation is the one of Example 5, in which the crystalline water content of the levamlodipine besylate hydrate composition is 5.3%.

[0099] In this experiment, the LD50 is measured by two administration routes. Oral administration and intraperitoneal administration of levamlodipine besylate hydrate composition tablets, grinding the tablets into uniform fine powder, and then using 10% gum Arabic to prepare two groups of liquid mixtures at a suitable concentration for oral administration and intraperitoneal administration. Animals: Kunming mice, half male and half male, each weighing 18-28 g, 10 animals in each group, fasted for 16 hours before use. Dosage: Oral administration: 1.2605, 1.0714, 0.910, 0.7141, 0.6580 (g/kg), intraperitoneal administration: 0.7743, 0.6582, 0.5594, 0.4755, 0.4042 (g/kg) dose gradient: 0.85. Route of administration: experimental methods of oral administration and intraperitoneal administration, 100 mice are selected, fasted for 16 hours, and randomly divided into 10 groups. Oral administration to the first 5 groups, and the dose being 0.35 ml/10 g; intraperitoneal administration to the latter 5 groups, and the dose being 0.2 ml/10 g. Observation of indicators: The animals are observed for 7 days after drug administration, and their toxic reactions and deaths are recorded. Animals display closed eyes, lethargy, sagging skin and sparse fur after 0 minutes of administration, and begin to die after 1 hour. Symptoms before death are balance disorders and convulsions etc., no significant changes are seen in postmortem examination.

TABLE-US-00011 TABLE 11 LD50 Results of Oral Administration (Bliss Method) Number Animal Daily Deaths (7 of Death Experimental Last Regression Dose Logarithmic Number days) Deaths Rate Probability Probability Probability mg/kg dose (number) |1|2|3|4|5|6|7| (number) (%) Unit Unit Unit Deviation 1260.5 3.1 10 |10|0|0|0|0|0|0| 10 100 6.96 7.18 7.18 0.000065 1071.4 2.03 10 |0|8|0|0|0|0|0| 8 80 5.84 6.14 6.14 0.000031 910 7 2.96 10 |0|6|0|0|0|0|0| 6 60 5.25 5.09 5.09 −0.000004 774.1 2.89 10 |0|2|0|0|0|0|0| 2 20 4.16 4.05 4.05 −0.000038 658.0 2.82 10 |0|0|0|0|0|0|0| 0 0 3.04 3.01 3.01 −0.000072

[0100] Significance index G=0.2018 X50=2.9531 SX=0.0164 G is relatively small.

[0101] Heterogeneity check Ch2=1.07 Ch2.05=7.82 Sb=3.3878 No heterogeneity.

[0102] Converted into the composition of levamlodipine besylate hydrate (calculated in levamlodipine) is LD50=24.925±1.7542 mg/kg.

TABLE-US-00012 TABLE 12 LD of Intraperitoneal Administration, Results (Bliss Method) Daily Animal Death(7 Number of Death Experimental Last Regression Dose Logarithmic Number Days) Deaths Rate probability probability Probability mg/kg dose (Number) |1|2|3|4|5|6|7| (Number) (%) unit Unity Unit deviation 774.3 2.89 10 |10|0|0|0|0|0|0| 10 100 6.96 6.73 6.73 0.000813 658.2 2.82 10 |0|7|0|0|0|0|0| 7 70 5.52 5.77 5.77 0.000416 559 4 2.75 10 |0|4|0|0|0|0|0| 4 40 4.75 4.82 4.82 0.0000(5 475.5 2.68 10 |0|2|0|0|0|0|0| 2 20 4.16 3.86 3.86 0.000378 404.2 2.61 10 |0|0|0|0|0|0|0| 0 0 3.04 2.91 2.91 0.000774

[0103] Significance index G=0.2002 X50=2.7612 SX=0.0172 G is relatively small and has been omitted.

[0104] Heterogeneity check Ch2=1.51 Ch2.05=7.82 Sb=3.3.0947 No heterogeneity.

[0105] Converted into the levamlodipine besylate hydrate composition (calculated in levamlodipine) is LD50=16.0212±1.3774 mg/kg.

Example 12 Pharmacokinetics of Sustained-Release Formulation

[0106] Please see Example 8 for the drug formulation of the fast-sustained-release formulation of the levamlodipine besylate hydrate composition used in this experiment. The purpose of this experiment is to study the pharmacokinetic behavior of the sustained-release tablets of the levamlodipine besylate hydrate composition (test preparation, dosage 10 mg/tablet,) in Beagle dogs, and to investigate whether it has the pharmacokinetic characteristic of fast and sustained dual release. The relative bioavailability in Beagle dogs is evaluated by using the solution of levamlodipine besylate hydrate composition prepared by the method described in Example 1 of the present invention as a reference preparation. Five adult and healthy Beagle dogs, all male, are divided into two groups for a single-dose administration experiment. The test and reference preparations are orally administered to the Beagle dogs on an empty stomach at a dose of 10 mg/dog, and plasma samples are collected at different time points. Liquid chromatography-mass spectrometer is used to determine the concentration of the levamlodipine besylate hydrate composition (calculated in levamlodipine) in plasma, and the pharmacokinetic parameters are calculated. The following results are obtained:

TABLE-US-00013 TABLE 13 Pharmacokinetic Parameters After Single-Dose Oral Administration of Test and Reference Preparations of Levamlodipine Besylate Hydrate Composition (mean ± SD) Pharmacokitetic Test Reference Parameter Preparation Preparation T.sub.max(h)  7.3 ± 1.2  2.0 ± 0.0 C.sub.max(ng/ml) 47.4 ± 7.5  69.8 ± 19.7 AUC.sub.0-t(ng .Math. h/ml) 1035.3 ± 318.5 1204.0 ± 415.8 AUC.sub.0-∞(ng .Math. h/ml) 1131.0 ± 385.4 1281.0 ± 492.1 MRT.sub.(h) 19.3 ± 4.5 16.4 ± 3.5 t.sub.1/2(h) 12.3 ± 2.8 11.2 ± 2.8 F (%) 86.0%

[0107] After oral administration of 10 mg of the test and reference preparations of the levamlodipine besylate hydrate compositions to Beagle dogs, the mean plasma concentration-time curve is shown in FIG. 10.

[0108] The results show that after oral administration of the sustained-release tablets of the levamlodipine besylate hydrate compositions to Beagle dogs, the average peak plasma concentration of levamlodipine in the dogs is 47.4 ng/ml, which is lower than the peak concentration of 69.8 ng/ml of the reference preparation by 32%, indicating that the sustained-release tablets can effectively reduce the peak concentration of the fast-release preparation; The plasma concentration of the sustained-release tablets of the levamlodipine besylate hydrate 1 hour after administration is more than 20% of the peak concentration of the reference preparation, and the average peak time is 7.3 h, which is longer than the 2.0 h peak time of the reference preparation. It has the fast and sustained dual release characteristic, and can maintain a steady plasma concentration over a period of time; the bioavailability of the sustained-release tablets of the levamlodipine besylate hydrate composition relative to the reference preparation is 86.0%, indicating that the absorption degree of the two preparations in Beagle dogs is basically the same. Given that the gastrointestinal tract of dogs is relatively short and the drug excretion is relatively fast, it is expected that the sustained release effect and the degree of absorption of the sustained-release preparation in human trials will be better than those of dogs.