Pharmaceutical composition, preparation and metolazone lyophilized powder preparation, and preparation methods therefor and uses thereof

Abstract

Disclosed in the present invention are a pharmaceutical composition, preparation and metolazone lyophilized powder preparation, and preparation methods therefor and the uses thereof. The pharmaceutical composition is a metolazone inclusion complex and contains metolazone or a salt thereof and a β-cyclodextrin derivative, wherein the molecular molar ratio of the metolazone or the salt thereof to the β-cyclodextrin derivative is 1:2 to 1:8. In the present invention, it is unexpectedly found by means of experiments that metolazone can undergo inclusion with the β-cyclodextrin derivative, thereby greatly improving the water solubility of metolazone, and that the metolazone lyophilized injection prepared using the metolazone inclusion complex has a higher diuretic speed, a stronger diuretic effect and long-lasting efficacy compared with innovator tablets at the same dose, can rapidly exert a strong diuretic effect after administration, is particularly suitable for emergency treatment of patients with acute heart failure accompanied by loop diuretic resistance, and has a significant value in clinical application.

Description

Pharmaceutical composition, preparation, metolazone lyophilized powder preparation, and preparation method and use thereof

[0001] This application claims the benefit of Chinese Patent Application No. 2023116426906, filed December 1, 2023. This application incorporates the entirety of the aforementioned Chinese Patent Application. Technical Field

[0002] The present invention relates to a pharmaceutical composition, a preparation, a metolazone lyophilized powder preparation, and a preparation method and use thereof. Background Art

[0003] Metolazone (CAS: 17560-51-9) is a quinazoline diuretic used to treat edema caused by congestive heart failure and hypertension. Metolazone has a strong, stable, and long-lasting diuretic effect.

[0004] Metolazone primarily acts on the distal convoluted tubule to inhibit sodium ion reabsorption and increase urine volume. This differs from loop diuretics, which act on the thick ascending limb of the loop of Henle. Therefore, the diuretic effects of the two can be complementary and additive. Furthermore, loop diuretic therapy leads to enhanced sodium ion reuptake in the distal convoluted tubule, increasing metolazone's sensitivity to its effects. Therefore, metolazone and loop diuretics (such as furosemide) exhibit synergistic effects in their pharmacodynamic mechanisms. Clinical application has shown that the combined diuretic effect of metolazone and furosemide is superior to either drug alone and can significantly improve the diuretic effect in patients with furosemide resistance. For example, the American Heart Failure Guidelines, "2022 AHA / ACC / HFSA Guideline for the Management of Heart Failure," recommend that metolazone be used as an add-on therapy after loop diuretic resistance develops.

[0005] Metolazone is an important treatment for acute decompensated heart failure (ADHF) after resistance to loop diuretics. Metolazone has a strong and long-lasting diuretic effect, and its mechanism complements that of loop diuretics, effectively resolving resistance to diuretics such as furosemide. Furthermore, unlike hydrochlorothiazide, a drug with a similar mechanism, metolazone does not reduce renal blood flow and glomerular filtration rate, making it suitable for patients with severe renal impairment. However, metolazone's current formulation limits its clinical application. As an oral preparation, it takes about an hour to achieve a satisfactory diuretic effect, making it ineffective for critically ill heart failure patients. For some heart failure patients with severe edema, oral drug absorption is impaired due to edema involving the intestines, further impacting the diuretic and de-edema effects of the drug.

[0006] Intravenous metolazone preparations can solve the above-mentioned problems in clinical applications, but the extremely low solubility of metolazone in water (0.02 mg / ml) limits the development of its intravenous preparations.

[0007] In order to solve this problem, U.S. Patent US 5124152A adds a large amount of organic solvents such as ethanol, propylene glycol, polyethylene glycol, etc. to the preparation prescription. A large amount of organic solvents not only causes irritation at the administration site, but also brings clinical medication risks such as hemolysis and allergies; U.S. Patent US 9427398 B2 discloses a solution system containing N, N-dimethylacetamide (DMAC) cosolvent to prepare metolazone injection. However, the DMAC used in this patent has not passed the approval of the State Food and Drug Administration for pharmaceutical excipients, and there are unknown safety risks in clinical use; U.S. Patent US2020 / 0179386A1 uses homogeneous dispersion technology to prepare metolazone with vegetable oil, phospholipid emulsifiers, Tween-80 and poloxamer and other emulsifiers into a fat emulsion preparation, which can be used for intravenous administration, but the patent formulation contains a high content of Tween-80 and poloxamer, and intravenous administration has a higher risk of hemolysis and allergies.

[0008] In summary, metolazone is a poorly water-soluble drug, and only some comparative documents describe compositions and methods for its preparation into injections. However, there are still risks to drug safety. Moreover, these existing technologies do not address the poor stability of metolazone's intravenous injection preparations. Therefore, no metolazone intravenous injection preparation that meets drug quality requirements has been marketed. Summary of the Invention

[0009] In order to overcome the above-mentioned problems of drug safety and poor stability of the cosolvents or coemulsifiers in the metolazone injections in the prior art, a pharmaceutical composition, preparation, metolazone lyophilized powder preparation and preparation method and use thereof are provided. The metolazone inclusion compound prepared by the present invention has good water solubility and safe ingredients. The lyophilized powder injection further prepared has good stability when used as an injection dosage form, which makes it possible to directly prepare metolazone into dosage forms such as injections for injection into the body, and is of great value for clinical treatment.

[0010] The objectives of the present invention are achieved through the following technical solutions.

[0011] In a first aspect, the present invention provides a pharmaceutical composition, which is a metolazone inclusion compound containing metolazone or a salt thereof and a β-cyclodextrin derivative, wherein the molecular molar ratio of the metolazone or a salt thereof to the β-cyclodextrin derivative is 1:2 to 1:8.

[0012] In the present invention, the metolazone or its salt in the metolazone inclusion compound is encapsulated in the cavity of the β-cyclodextrin derivative.

[0013] In the present invention, the salt of metolazone may be a pharmaceutically acceptable salt thereof.

[0014] In the present invention, the metolazone salt is generally a salt formed by metolazone and an inorganic base or an organic base.

[0015] The inorganic base can be selected from one or more inorganic bases containing sodium, potassium, magnesium, calcium and aluminum.

[0016] The organic base can be selected from one or more of methylamine, ethylamine, ethanolamine, lysine, arginine and ornithine.

[0017] In the present invention, the β-cyclodextrin derivative is conventionally selected in the art, preferably one or more of sulfobutyl ether β-cyclodextrin sodium, hydroxypropyl β-cyclodextrin and methyl β-cyclodextrin.

[0018] In the present invention, the molecular molar ratio of the metolazone or its salt to the β-cyclodextrin derivative is preferably 1:3 to 1:6, more preferably 1:4.

[0019] In the present invention, the β-dextrin derivative in the metolazone inclusion complex is a type of pharmaceutical excipient with low toxicity, which has been approved for oral or injection administration both at home and abroad. In particular, sulfobutyl ether β-cyclodextrin sodium is less toxic than other β-cyclodextrin derivatives and is safer for clinical use.

[0020] In the present invention, too low or too high an amount of the β-cyclodextrin derivative is not conducive to the formation of the inclusion complex. In particular, when the amount of the β-cyclodextrin derivative is too low, the inclusion of the drug cannot be completed, and precipitation will occur during re-dissolution. When the amount of the β-cyclodextrin derivative is too high, it exceeds the necessary amount, the manufacturing cost is greatly increased, and the effect has reached saturation at this time.

[0021] In some embodiments of the present invention, the metolazone inclusion compound may further comprise at least one pharmaceutically acceptable carrier, excipient or diluent, and may be combined with other active ingredients if necessary.

[0022] In a second aspect, the present invention provides a method for preparing a pharmaceutical composition, wherein the pharmaceutical composition is as described above, comprising any of the following methods:

[0023] Method 1: dissolving the metolazone or its salt and the β-cyclodextrin derivative in a first solvent and stirring until the solution becomes clear and transparent.

[0024] Method 2: dissolving the metolazone or its salt in a second solvent to obtain a metolazone solution, dissolving the β-cyclodextrin derivative in a third solvent to obtain a β-cyclodextrin derivative solution, and then mixing the two and stirring until the solution is clear and transparent.

[0025] Method 3: Mix the β-cyclodextrin derivative with water, then mix with the metolazone or its salt, add an alkaline pH regulator, stir until clear, and then add an acidic pH regulator to neutralize.

[0026] Wherein, the first solvent can be selected from one or more of water, ethanol, methanol, propanol, isopropanol, acetone, ethylene glycol, propylene glycol, glycerol, ethyl acetate, dichloromethane, tetrahydrofuran and tert-butanol. When the first solvent is water, the first solvent also contains an alkaline pH regulator, and after the "stirring until clear and transparent" step, an acidic pH regulator is added for neutralization; preferably, the first solvent is selected from: ethanol, a mixed solvent of ethanol and water, tert-butanol, and a mixed solvent of tert-butanol and water; more preferably, the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 1:3 to 3:1, and the volume ratio of tert-butanol to water in the mixed solvent of tert-butanol and water is 1:3 to 2:1.

[0027] The second solvent may be selected from one or more of water, ethanol, anhydrous ethanol, methanol, propanol, isopropanol, acetone, ethylene glycol, propylene glycol, glycerol, ethyl acetate, dichloromethane, tetrahydrofuran and tert-butanol. Preferably, the second solvent is selected from anhydrous ethanol and / or tert-butanol. When the second solvent and the third solvent are both water, the second solvent further contains a pH alkaline regulator, and after the step of "stirring until clear and transparent", an acidic pH regulator is added for neutralization.

[0028] Wherein, the third solvent can be selected from one or more of water, ethanol, methanol, propanol, isopropanol, acetone, ethylene glycol, propylene glycol, glycerol, ethyl acetate, dichloromethane, tetrahydrofuran and tert-butanol; preferably, the third solvent is selected from: water, ethanol, a mixed solvent of ethanol and water, tert-butanol, and a mixed solvent of tert-butanol and water; more preferably, the volume ratio of ethanol and water in the mixed solvent of ethanol and water is 1:3 to 3:1, and the volume ratio of tert-butanol and water in the mixed solvent of tert-butanol and water is 1:3 to 2:1.

[0029] The stirring temperature may be 4°C to 60°C, for example, 4°C, 10°C, 25°C, 40°C or 60°C.

[0030] The stirring time may be 30-120 min, for example 30 min, 60 min or 120 min.

[0031] The dosage of metolazone can be controlled so that the mass volume percentage of metolazone in the solution is 0.05-0.3%; the mass volume percentage refers to the ratio of the mass (g) of metolazone in the solution to the total volume (ml) of the solution.

[0032] In the present invention, those skilled in the art know that after the "stirring until clear and transparent", a metolazone inclusion compound solution is obtained. Generally speaking, a subsequent drying step may be included, and after drying, a metolazone inclusion compound powder is obtained.

[0033] The drying method may be vacuum rotary evaporation, spray drying or freeze drying.

[0034] In the present invention, the alkaline pH regulator can be a conventional pharmaceutical alkaline pH regulator in the art, and can be selected from one or more of sodium hydroxide, sodium carbonate, sodium bicarbonate, triethanolamine, ammonia water, potassium hydroxide, potassium sulfate, tromethamine, meglumine and basic amino acids, preferably sodium hydroxide.

[0035] In the present invention, the amount of the alkaline pH regulator used can be an amount that can be stirred to a clear and transparent solution, and can generally be used to adjust the solution to a pH of 11 to 12.

[0036] In the present invention, the acidic pH regulator can be a conventional pharmaceutical acidic pH regulator in the art, and can be selected from one or more of citric acid, hydrochloric acid, sulfuric acid, acetic acid, tartaric acid, lactic acid, malic acid, phosphoric acid, fumaric acid, maleic acid, succinic acid and acidic amino acids, preferably citric acid.

[0037] In the present invention, the acidic pH regulator is used in an amount sufficient to neutralize the solution, and generally can be used to adjust the pH of the solution to 6-8.

[0038] In a third aspect, the present invention provides a pharmaceutical preparation comprising the aforementioned metolazone inclusion compound or the pharmaceutical composition prepared by the aforementioned preparation method, and at least one pharmaceutically acceptable carrier, excipient or diluent, or other active ingredient.

[0039] Preferably, the pharmaceutical composition is in the form of an infusion, a liquid injection, a powder injection, a tablet, a capsule, a granule, a dispersible tablet, an oral solution or a syrup.

[0040] The metolazone inclusion complex of the present invention can be administered parenterally in the form of a sterile injectable aqueous solution. Such parenteral administration includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, or other methods of administration known in the art. The metolazone inclusion complex for parenteral administration may also contain a pharmaceutically acceptable carrier and solvent, including water, Ringer's solution, isotonic sodium chloride solution, or glucose solution. Formulations for parenteral administration may include infusions, aqueous injections, powder injections, and the like.

[0041] The metolazone inclusion complex of the present invention can be administered as an oral preparation, which may contain a pharmaceutically acceptable solid carrier or liquid carrier. Solid carriers generally include diluents, flavoring agents, lubricants, binders, disintegrants, and the like commonly used in pharmacy. Liquid carriers include water, ethanol, propylene glycol, glycerol, corn syrup, and the like. Liquid preparations may also contain pharmaceutically acceptable sweeteners, flavoring agents, preservatives, and the like. Solid preparations may include tablets, capsules, granules, and dispersible tablets; liquid preparations may include oral solutions and syrups.

[0042] In a fourth aspect, the present invention further provides a metolazone lyophilized powder preparation, comprising the aforementioned metolazone inclusion compound or a pharmaceutical composition prepared by the aforementioned preparation method.

[0043] In the present invention, the lyophilized powder preparation may further include a lyoprotectant, such as one or more of mannitol, inositol, sorbitol, lactose, sucrose, glucose, trehalose, maltose, dextran, glycine, PVP and polyethylene glycol.

[0044] In the present invention, the lyophilized powder preparation may further include a pH adjuster, such as one or more of sodium hydroxide, hydrochloric acid, citric acid, tartaric acid, phosphoric acid, sodium bicarbonate and amino acids.

[0045] In a fifth aspect, the present invention provides a method for preparing a metolazone lyophilized powder preparation, wherein the metolazone lyophilized powder preparation adopts the aforementioned metolazone lyophilized powder preparation, and comprises the following steps: when the pharmaceutical composition is in powder form, re-dissolving and filtering, or filtering the metolazone inclusion complex solution obtained by "stirring until clear and transparent"; and then drying.

[0046] The reconstitution solvent may be water, preferably water for injection. Preferably, the amount of water is 0.4-2 times the volume of the metolazone inclusion solution, and the volume of the metolazone inclusion solution is the volume of the aforementioned first solvent or the total volume of the second solvent and the third solvent.

[0047] The filtration method is conventional in the art, preferably membrane filtration.

[0048] The drying method may be freeze drying, spray drying or reduced pressure drying, preferably freeze drying.

[0049] In the present invention, preferably, a freeze-drying protective agent may be added to dissolve the solution before filtering.

[0050] The freeze-dried powder for injection prepared by using the metolazone inclusion compound of the present invention can significantly improve the stability of the metolazone preparation.

[0051] In a sixth aspect, the present invention provides a use of a metolazone inclusion compound or a metolazone lyophilized powder preparation in the preparation of a diuretic drug, wherein the diuretic drug can be used to treat edema and hypertension.

[0052] Wherein, the metolazone inclusion compound is the aforementioned metolazone inclusion compound, or the metolazone inclusion compound prepared by the aforementioned preparation method,

[0053] Wherein, the metolazone lyophilized powder preparation is a metolazone lyophilized powder preparation obtained by the aforementioned metolazone lyophilized powder preparation or the aforementioned preparation method.

[0054] On the basis of conforming to the common sense in this field, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0055] The reagents and raw materials used in the present invention are commercially available.

[0056] The positive progress effect of the present invention is:

[0057] (1) Through experiments, it was unexpectedly discovered that metolazone or its salts can form an inclusion complex with β-cyclodextrin derivatives, thereby greatly improving the water solubility of metolazone and overcoming the disadvantage that metolazone is difficult to prepare into a water-soluble preparation, especially a liquid preparation for intravenous injection; the inclusion complex increases the water solubility of metolazone by more than 100 times, and the oral preparation prepared with it also has the characteristics of good dissolution and high bioavailability, which is more conducive to clinical application.

[0058] (2) The metolazone lyophilized injection prepared using the metolazone inclusion compound of the present invention has a faster diuretic rate, a stronger diuretic effect, and a long-lasting efficacy compared to the original tablets of the same dose. It can quickly exert a strong diuretic effect after administration and is particularly suitable for emergency treatment of patients with acute heart failure accompanied by loop diuretic resistance, and has significant clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 is an X-ray diffraction pattern, wherein part a of Figure 1 is an X-ray diffraction pattern of the metolazone powder used as a raw material in Example 4; part b of Figure 1 is an X-ray diffraction pattern of the sulfobutyl ether β-cyclodextrin powder used as a raw material in Example 4; part c of Figure 1 is an X-ray diffraction pattern of a physical mixture of metolazone powder and sulfobutyl ether β-cyclodextrin; and part d of Figure 1 is an X-ray diffraction pattern of the metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex prepared in Example 4.

[0060] FIG2 shows the real-time urine production rate (mean±SD, μL / min) of Experimental Example 3.

[0061] FIG3a shows the diuretic onset time of Experimental Example 3 (mean±SD, min).

[0062] FIG3 b shows the time to peak diuretic effect in Experimental Example 3 (mean±SD, min).

[0063] FIG3 c shows the peak increase in urine production rate of Experimental Example 3 (mean±SD, min).

[0064] FIG. 4 shows the cumulative urine volume after administration in Test Example 3 (mean±SD, mL).

[0065] FIG5 shows the increase in the cumulative urine volume of Experimental Example 3 compared with the negative control group (mean±SD, %). DETAILED DESCRIPTION

[0066] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0067] Example 1

[0068] 2.53 g of hydroxypropyl β-cyclodextrin and 100 mg of metolazone (the molar ratio of metolazone to hydroxypropyl β-cyclodextrin is 1:6) were weighed separately, added to 50 ml of ethanol, stirred at 25 ° C for 30 minutes until clear and transparent, and dried by vacuum rotary evaporation to obtain a dry solid, that is, the metolazone hydroxypropyl β-cyclodextrin inclusion complex.

[0069] Example 2

[0070] 1.78 g of sodium sulfobutyl ether β-cyclodextrin was dissolved in 40 ml of water to obtain solution A. 100 mg of metolazone was dissolved in 60 ml of anhydrous ethanol to obtain solution B (the molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin was 1:3). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 40°C for 60 minutes until clear and transparent. The dried solid was obtained by rotary evaporation under reduced pressure to obtain the metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex.

[0071] Example 3

[0072] 1.69 g of hydroxypropyl β-cyclodextrin and 100 mg of metolazone (the molar ratio of metolazone to hydroxypropyl β-cyclodextrin is 1:4) were weighed separately, added to 100 ml of tert-butanol, stirred at 10°C for 120 minutes until clear and transparent, and evaporated under reduced pressure to a dry solid to obtain metolazone hydroxypropyl β-cyclodextrin inclusion complex.

[0073] Example 4

[0074] 5.92 g of sodium sulfobutyl ether β-cyclodextrin was dissolved in 40 ml of water to obtain solution A. 250 mg of metolazone was dissolved in 60 ml of anhydrous ethanol to obtain solution B (the molecular molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin was 1:4). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 25°C for 60 minutes until clear and transparent. The mixture was evaporated under reduced pressure to a dry solid to obtain a metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex.

[0075] Example 5

[0076] Weigh 0.59 g of sodium sulfobutyl ether β-cyclodextrin and dissolve it in 25 ml of water to obtain solution A. Weigh 50 mg of metolazone and dissolve it in 75 ml of anhydrous ethanol to obtain solution B (the molecular molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin is 1:2). Then slowly add solution B to solution A and mix well. Stir at 4°C for 60 minutes until clear and transparent. Vacuum evaporate to a dry solid to obtain metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex.

[0077] Example 6

[0078] 4.74 g of sodium sulfobutyl ether β-cyclodextrin was dissolved in 75 ml of water to obtain solution A. 100 mg of metolazone was dissolved in 25 ml of anhydrous ethanol to obtain solution B (the molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin was 1:8). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 60°C for 60 minutes until clear and transparent. The mixture was evaporated under reduced pressure to a dry solid to obtain a metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex.

[0079] Example 7

[0080] 8.88 g of sodium sulfobutyl ether β-cyclodextrin was dissolved in 50 ml of water to obtain solution A. 300 mg of metolazone was dissolved in 50 ml of anhydrous ethanol to obtain solution B (the molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin was 1:5). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 25°C for 60 minutes until clear and transparent. The mixture was evaporated under reduced pressure to a dry solid to obtain a metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex.

[0081] Example 8

[0082] Weigh 2.37 g of sodium sulfobutyl ether β-cyclodextrin, add it to 98 ml of water for injection and stir to dissolve, then add 100 mg of metolazone and stir and disperse it thoroughly (the molecular molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin is 1:4), and at the same time, add 0.7 ml of 4 M sodium hydroxide solution to adjust the pH of the solution to 11. Continue stirring until a clear and transparent aqueous solution of metolazone is obtained, and then add 0.9 ml of 1 M citric acid solution to adjust the pH of the solution to 7 to obtain a clear, transparent, stable and non-precipitated pH neutral metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex solution.

[0083] Example 9

[0084] 3.56 g of sodium sulfobutyl ether β-cyclodextrin was dissolved in 33.3 ml of water to obtain solution A. 200 mg of metolazone was dissolved in 66.7 ml of tert-butanol to obtain solution B (the molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin was 1:3). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 25°C for 60 minutes until clear and transparent. The mixture was evaporated under reduced pressure to a dry solid to obtain a metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex.

[0085] Example 10

[0086] 4.44 g of sodium sulfobutyl ether β-cyclodextrin was dissolved in 40 ml of water to obtain solution A. 250 mg of metolazone was dissolved in 60 ml of anhydrous ethanol to obtain solution B (the molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin was 1:3). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 25°C for 60 minutes until clear and transparent. The mixture was evaporated under reduced pressure to a dry solid to obtain a metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex.

[0087] Example 11

[0088] 1.43 g of methyl β-cyclodextrin was dissolved in 40 ml of water to obtain solution A. 100 mg of metolazone was dissolved in 60 ml of anhydrous ethanol to obtain solution B (the molar ratio of metolazone to methyl β-cyclodextrin molecules was 1:4). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 25°C for 60 minutes until clear and transparent. The mixture was evaporated under reduced pressure to a dry solid to obtain metolazone methyl β-cyclodextrin sodium inclusion complex.

[0089] Example 12

[0090] Metolazone lyophilized powder for injection

[0091] The metolazone hydroxypropyl β-cyclodextrin inclusion compound prepared in Example 1 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capping.

[0092] Example 13

[0093] Metolazone lyophilized powder for injection

[0094] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex prepared in Example 2 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capped.

[0095] Example 14

[0096] Metolazone lyophilized powder for injection

[0097] The metolazone hydroxypropyl β-cyclodextrin inclusion complex prepared in Example 3 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capping.

[0098] Example 15

[0099] Metolazone lyophilized powder for injection

[0100] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex prepared in Example 4 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capped.

[0101] Example 16

[0102] Metolazone lyophilized powder for injection

[0103] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex solution prepared in Example 5 was filtered through a filter membrane, divided into vials, placed in a freeze dryer, freeze-dried, and capped.

[0104] Example 17

[0105] Metolazone lyophilized powder for injection

[0106] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex solution prepared in Example 6 was filtered through a filter membrane, divided into vials, placed in a freeze dryer, freeze-dried, and capped.

[0107] Example 18

[0108] Metolazone lyophilized powder for injection

[0109] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex prepared in Example 7 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capped.

[0110] Example 19

[0111] Metolazone lyophilized powder for injection

[0112] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex solution prepared in Example 8 was filtered through a filter membrane, divided into vials, placed in a freeze dryer, freeze-dried, and capped.

[0113] Example 20

[0114] Metolazone lyophilized powder for injection

[0115] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex prepared in Example 9 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capped.

[0116] Example 21

[0117] Metolazone lyophilized powder for injection

[0118] The metolazone sulfobutyl ether β-cyclodextrin sodium inclusion complex prepared in Example 10 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capped.

[0119] Example 22

[0120] Metolazone lyophilized powder for injection

[0121] The metolazone methyl β-cyclodextrin inclusion complex prepared in Example 11 was added into 100 ml of water for injection and redissolved until clear and transparent. The mixture was filtered through a filter membrane and divided into vials. The vials were placed in a freeze dryer for freeze drying and capped.

[0122] Comparative Example 1

[0123] According to Example 2 in US Pat. No. 5,124,152A, a metolazone injection was prepared, containing 1.17 mg / ml of metolazone, 10% by volume of 95% ethanol solution, and 40% by volume of propylene glycol.

[0124] Comparative Example 2

[0125] According to Example 3 in US Pat. No. 9,427,398 B2, a metolazone injection was prepared containing 150 mg of metolazone, 0.1 mL of dimethylacetamide (DMAC), and 9.9 mL of polyethylene glycol 400.

[0126] Comparative Example 3

[0127] Example 26 in U.S. Patent US 2020 / 0179386 A1 document prepares a metolazone fat emulsion injection containing 1 mg / ml metolazone, 20% soybean oil, 12 mg / ml soybean lecithin, 2.25% glycerol, and 3% poloxamer.

[0128] Comparative Example 4

[0129] Weigh 0.6 g of sodium sulfobutyl ether β-cyclodextrin and dissolve it in 40 ml of water to obtain solution A. Weigh 100 mg of metolazone and dissolve it in 60 ml of anhydrous ethanol to obtain solution B (the molecular molar ratio of metolazone to sodium sulfobutyl ether β-cyclodextrin is 1:1). Then, slowly add solution B to solution A and mix well. Stir at 40°C for 60 minutes until it becomes clear and transparent, and evaporate under reduced pressure to obtain a dry solid.

[0130] Comparative Example 5

[0131] 1.29 g of 2-hydroxypropyl α-cyclodextrin was dissolved in 40 ml of water to obtain solution A. 100 mg of metolazone was dissolved in 60 ml of anhydrous ethanol to obtain solution B (the molar ratio of metolazone to 2-hydroxypropyl α-cyclodextrin was 1:4). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 40°C for 60 minutes until clear and transparent, and then evaporated under reduced pressure to obtain a dry solid.

[0132] Comparative Example 6

[0133] 1.89 g of hydroxypropyl γ-cyclodextrin was dissolved in 40 ml of water to obtain solution A. 100 mg of metolazone was dissolved in 60 ml of anhydrous ethanol to obtain solution B (the molar ratio of metolazone to hydroxypropyl γ-cyclodextrin was 1:4). Solution B was then slowly added to solution A and mixed. The mixture was stirred at 40°C for 60 minutes until clear and transparent, and then vacuum evaporated to obtain a dry solid.

[0134] Effect embodiment

[0135] The X-ray diffraction pattern of the inclusion complex prepared in Example 4 shows that the characteristic crystal peaks of metolazone in the sample have disappeared, indicating that it has been successfully included with the β-cyclodextrin derivative (Figure 1). Specifically, Figure 1 (a) shows the X-ray diffraction pattern of the metolazone powder used as the raw material in Example 4; Figure 1 (b) shows the X-ray diffraction pattern of the sulfobutyl ether β-cyclodextrin powder used as the raw material in Example 4; Figure 1 (c) shows the X-ray diffraction pattern of the physical mixture of metolazone powder and sulfobutyl ether β-cyclodextrin; and Figure 1 (d) shows the X-ray diffraction pattern of the sodium inclusion complex of metolazone sulfobutyl ether β-cyclodextrin prepared in Example 4.

[0136] Among them, the metolazone powder X-ray diffraction pattern in part a of Figure 1 has characteristic peaks at 7.36, 19.6 and 25.89;

[0137] The X-ray diffraction pattern of sulfobutyl ether β-cyclodextrin powder in part b of Figure 1 also has a peak at around 20;

[0138] The physical mixture of metolazone powder and sulfobutyl ether β-cyclodextrin in part c of Figure 1 can be seen to be a simple superposition of metolazone and sulfobutyl ether β-cyclodextrin;

[0139] However, the inclusion complex prepared in Example 4 in part d of FIG1 shows that the characteristic crystal peak of metolazone disappears, indicating that the inclusion result indicates that metolazone is encapsulated in the cavity of the β-cyclodextrin derivative.

[0140] Test Example 1

[0141] Inclusion complex solubility experiment

[0142] The inclusion compounds prepared in Examples 1-11 were added to 100 ml of physiological saline and stirred, and all of them quickly dissolved to become clear and transparent, and were stable without precipitation after being placed at room temperature, as shown in Table 1 below. This shows that the inclusion compounds were successfully prepared.

[0143] The solid prepared in Comparative Example 4-6 was added to 100 ml of physiological saline and stirred, but no clear solution was obtained. After standing at room temperature, particles precipitated, indicating that no stable inclusion complex was formed.

[0144] Table 1

[0145] Test Example 2 Preparation Stability Test

[0146] The metolazone lyophilized powder for injection prepared in Examples 12-22 and the metolazone injection prepared in Comparative Examples 1, 2, and 3 (packed in vials and sealed with caps) were subjected to high temperature testing and strong light irradiation testing in accordance with the guidelines for stability testing of pharmaceutical preparations in the Chinese Pharmacopoeia.

[0147] High temperature test conditions: Place the test sample at 60°C for 10 days, take samples on the 5th and 10th days, and detect changes in the preparation properties, main drug content and related substances.

[0148] Related substances are impurities in a drug that are produced from the same source and process as the main ingredient and have similar or related structures and properties. They can be byproducts of the drug production process or degradation products during storage, and are different from ordinary impurities.

[0149] Strong light irradiation test conditions: Place the test sample in a lighting device at 4500lx±500x for 10 days. Take samples on the 5th and 10th days to detect changes in the preparation properties, main drug content and related substances.

[0150] The test samples prepared in the examples that have undergone stability tests were reconstituted with normal saline for injection to 1 mg / ml according to the labeled dosage of the preparation. The content of the main drug and related substances were detected by high performance liquid chromatography (HPLC). The specific detection method is as follows:

[0151] An Agilent 1260 liquid chromatograph was used, equipped with a G7115A UV detector at 230 nm. The chromatographic column was an Agilent ZORBAX Eclipse XDB C18 analytical column (4.6 × 150 mm, 5 μm). Mobile phase A consisted of 95% aqueous solution containing 0.1% formic acid and 5% methanol; mobile phase B consisted of 90% acetonitrile containing 0.05% formic acid and 10% methanol. The flow rate was 1.0 mL / min, the column temperature was 30°C, and the injection volume was 15 μL. The gradient elution is shown in Table 2 below.

[0152] Table 2

[0153] The results of the high temperature test and strong light irradiation test are shown in the following table:

[0154] Table 3

[0155] The result data showed that under the high temperature and light conditions of this test, the freeze-dried powder injection preparations prepared in Examples 12-22 of the present invention were stable, with no significant change in properties, and very little change in content and related substances. In the high temperature test, the main drug content of the preparations prepared in the comparative examples decreased slightly to varying degrees, while the related substances increased significantly. Among them, the fat emulsion prepared in Comparative Example 3 showed oil floating and stratification. Under strong light irradiation conditions, the main drug content of the comparative examples decreased significantly, while the related substances increased significantly, and the stability of the preparations was seriously damaged.

[0156] Test Example 3 Diuretic Effect Test

[0157] The experiment involved 24 male rats weighing 280-320 g, randomly divided into three groups (n=8). They were fed a standard diet with free access to water during the feeding period and fasted overnight before the experiment. The specific experimental procedures are as follows:

[0158] 1) Anesthesia was administered by intraperitoneal injection of 50 mg / kg sodium pentobarbital. A skin incision was made on the right hind limb, and a 0.9 mm OD PE catheter was placed in the femoral vein at a depth of 15 mm to maintain anesthesia. A midline abdominal incision was made, and a 1.2 mm OD silicone cannula was inserted into the bladder at a depth of 5 mm. This cannula was secured with bio-glue for urine collection. A 1.2 mm OD silicone cannula was placed intraperitoneally through the incision for intraperitoneal fluid replacement. The incision was covered with saline-soaked gauze.

[0159] 2) Maintenance of anesthesia and fluid replacement: Anesthesia was maintained by continuous infusion of 8 mg / mL sodium pentobarbital via a silicone tube connected to a PE catheter using a microsyringe pump (Baoding Lange) at a rate of 0.5 mL / h. Fluid replacement was achieved by continuous intraperitoneal infusion of normal saline via another silicone tube at a rate of 0.5 mL / h. This infusion was continued from the baseline period until the end of the experiment.

[0160] 3) Administration: After the baseline period, the intravenous group received a tail vein injection of the sample prepared in Example 15 at a dose of 2 mg / kg (0.1 mL / 100 g body weight); the oral group received a direct intragastric injection of a metolazone tablet suspension at a dose of 2 mg / kg (0.1 mL / 100 g body weight) via gastric wall puncture; and the control group received a tail vein injection of an equal volume of normal saline.

[0161] 4) Urine collection: Urine outflow from the bladder was collected using 2 ml centrifuge tubes for a total of 70 minutes during the baseline period, with 20 minutes each for the first three points and 10 minutes for the last point. Urine was collected every 10 minutes for the first hour after administration, and every 20 minutes from the second hour onwards, for a total of 420 minutes.

[0162] 5) The collected urine is weighed to calculate the urine volume, urine volume rate, cumulative urine volume and cumulative urine volume increase percentage. The urine volume rate is the ratio of the urine volume collected in the interval to the interval duration. The cumulative urine volume is the ratio of the urine volume collected in the interval to the interval duration. (ΣVm: drug administration group, cumulative urine volume (mL) per mouse after drug administration; Average cumulative urine volume of the control group after administration (mL).

[0163] Dosage groups: intravenous group (lyophilized powder prepared in Example 15, recorded as iv) and oral group (tablets, ig).

[0164] Control group: saline control group.

[0165] The results showed that after administration of the intravenous (iv) and oral (ig) groups, the urination rate of the animals was significantly increased compared with the saline control group. For details, see Figure 2 for the real-time rate of urine production (mean±SD, μL / min). The urination rate of the intravenous (iv) group increased to more than twice the baseline within 10 minutes and was maintained for a long time. It still had a certain diuretic effect 7 hours after administration. The baseline value of the urination rate was the total urine volume collected before administration / 70 minutes.

[0166] Further analysis revealed that the oral group (ig) had a relatively slow onset of effect, with an average onset time of 53.33 ± 12.11 minutes. The intravenous group (iv) experienced rapid onset of effect, approximately 5.33 times faster than the oral group (see Figure 3a for diuretic onset time (mean ± SD, min)). The oral group (ig) experienced a slow peak in efficacy, with an average time to peak of 151.67 ± 67.06 minutes. However, the intravenous group (iv) experienced a rapid peak in efficacy, with an average time to peak of 31.25 ± 16.15 minutes, approximately 4.85 times faster than the oral group (ig) (see Figure 3b for diuretic peak time (mean ± SD, min)). After administration, the average peak urination rate of the intravenous group (iv) increased by 19.69±5.51 μL / min compared with the baseline period. The average peak urination rate of the oral group (ig) was 12.87±2.98 μL / min. The intravenous group (iv) was approximately 1.53 times that of the oral group (ig). For details, see Figure 3c for the peak increase in urine production rate (mean±SD, min).

[0167] On the other hand, the cumulative urine volume after administration in the intravenous and oral groups was significantly higher than that in the control group, with the intravenous group showing a higher urine volume than the oral group (see Figure 4 for details, Cumulative Urine Volume after Administration (mean ± SD, mL)). The intravenous group showed the largest increase in cumulative urine volume compared to the control group 30 minutes after administration, while the oral group showed the largest increase in cumulative urine volume compared to the control group 140 minutes after administration. By 5 hours after administration, the increase in cumulative urine volume in both groups stabilized. After 7 hours, the cumulative urine volume in the intravenous and oral groups increased by 129.31 ± 55.35% and 74.59% ± 44.82% compared to the control group, respectively. The percentage increase in the intravenous group was 1.73 times that of the oral group (see Figure 5 for details, Cumulative Urine Volume Increase Compared to the Negative Control Group (mean ± SD,%)).

Claims

1. A pharmaceutical composition, characterized in that: The pharmaceutical composition is a metolazone inclusion compound, which contains metolazone or its salt and a beta-cyclodextrin derivative, and the molecular molar ratio of the metolazone or its salt to the beta-cyclodextrin derivative is 1:2-1:

8.

2. The pharmaceutical composition according to claim 1, characterized in that: The metolazone inclusion compound satisfies one or more of the following conditions: The metolazone salt is a salt of metolazone and an inorganic base or an organic base; preferably, the inorganic base is selected from one or more inorganic bases containing sodium, potassium, magnesium, calcium and aluminum; preferably, the organic base is selected from one or more of methylamine, ethylamine, ethanolamine, lysine, arginine and ornithine; The β-cyclodextrin derivative is selected from one or more of sulfobutyl ether β-cyclodextrin sodium, hydroxypropyl β-cyclodextrin and methyl β-cyclodextrin; The molecular molar ratio of the metolazone or its salt to the β-cyclodextrin derivative is 1:3 to 1:6, preferably 1:

4.

3. A method for preparing the pharmaceutical composition according to claim 1 or 2, characterized in that: The preparation method is any of the following: Method 1: dissolving the metolazone or its salt and the β-cyclodextrin derivative in a first solvent, and stirring until the solution is clear and transparent; Method 2: dissolving the metolazone or its salt in a second solvent to obtain a metolazone solution, dissolving the β-cyclodextrin derivative in a third solvent to obtain a β-cyclodextrin derivative solution, and then mixing the two and stirring until clear and transparent; Method 3: Mix the β-cyclodextrin derivative with water, then mix it with the metolazone or its salt, add an alkaline pH regulator at the same time, stir until clear and transparent, and then add an acidic pH regulator to neutralize it.

4. The preparation method according to any one of claims 1 to 3, characterized in that The preparation method meets one or more of the following conditions: The first solvent is selected from one or more of water, ethanol, methanol, propanol, isopropanol, acetone, ethylene glycol, propylene glycol, glycerol, ethyl acetate, dichloromethane, tetrahydrofuran and tert-butanol, wherein when the first solvent is water, the first solvent further contains an alkaline pH regulator, and after the step of "stirring until clear and transparent", an acidic pH regulator is further added for neutralization; preferably, the first solvent is selected from: ethanol, a mixed solvent of ethanol and water, tert-butanol, and a mixed solvent of tert-butanol and water; more preferably, the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 1:3 to 3:1, and the volume ratio of tert-butanol to water in the mixed solvent of tert-butanol and water is 1:3 to 2:1; The second solvent is selected from one or more of water, ethanol, anhydrous ethanol, methanol, propanol, isopropanol, acetone, ethylene glycol, propylene glycol, glycerol, ethyl acetate, dichloromethane, tetrahydrofuran and tert-butanol. Preferably, the second solvent is selected from anhydrous ethanol and / or tert-butanol. When the second solvent and the third solvent are both water, the second solvent further contains a pH alkaline regulator, and after the step of "stirring until clear and transparent", a pH acidic regulator is added for neutralization. The third solvent is selected from one or more of water, ethanol, methanol, propanol, isopropanol, acetone, ethylene glycol, propylene glycol, glycerol, ethyl acetate, dichloromethane, tetrahydrofuran and tert-butanol; preferably, the third solvent is selected from: water, ethanol, a mixed solvent of ethanol and water, tert-butanol, and a mixed solvent of tert-butanol and water; more preferably, the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 1:3 to 3:1, and the volume ratio of tert-butanol to water in the mixed solvent of tert-butanol and water is 1:3 to 2:

1.

5. The preparation method according to any one of claims 1 to 4, characterized in that The preparation method satisfies one or more of the following conditions: the stirring temperature is 4°C to 60°C, such as 4°C, 10°C, 25°C, 40°C or 60°C; The stirring time is 30-120 min, for example 30 min, 60 min or 120 min; The amount of metolazone is controlled so that the mass volume percentage of metolazone in the solution is 0.05-0.3%; The step of "stirring until clear and transparent" further includes a drying step to obtain metolazone inclusion compound powder; preferably, the drying method is vacuum rotary evaporation, spray drying or freeze drying.

6. The preparation method according to any one of claims 1 to 5, characterized in that The alkaline pH regulator is selected from one or more of sodium hydroxide, sodium carbonate, sodium bicarbonate, triethanolamine, ammonia water, potassium hydroxide, potassium sulfate, tromethamine, meglumine and basic amino acids, preferably sodium hydroxide; The acidic pH regulator is selected from one or more of citric acid, hydrochloric acid, sulfuric acid, acetic acid, tartaric acid, lactic acid, malic acid, phosphoric acid, fumaric acid, maleic acid, succinic acid and acidic amino acids, preferably citric acid; The alkaline pH regulator is used in an amount to adjust the solution pH to 11-12; The acidic pH regulator is used in an amount to adjust the solution pH to 6-8.

7. A pharmaceutical preparation, characterized in that The invention comprises the metolazone inclusion compound according to claim 1 or 2, or a pharmaceutical composition prepared by the preparation method according to any one of claims 3 to 6, and at least one pharmaceutically acceptable carrier, excipient or diluent, or other active ingredient; preferably, the pharmaceutical preparation is in the form of infusion, water injection, powder injection, tablet, capsule, granule, dispersible tablet, oral solution or syrup.

8. A metolazone lyophilized powder preparation, characterized in that: It comprises the metolazone inclusion compound as claimed in claim 1 or 2, or a pharmaceutical composition prepared by the preparation method as claimed in any one of claims 3 to 6; Preferably, the lyophilized powder preparation further comprises a lyoprotectant, such as one or more of mannitol, inositol, sorbitol, lactose, sucrose, glucose, trehalose, maltose, dextran, glycine, PVP and polyethylene glycol; Preferably, the lyophilized powder preparation further comprises a pH adjuster, such as one or more of sodium hydroxide, hydrochloric acid, citric acid, tartaric acid, phosphoric acid, sodium bicarbonate and amino acids.

9. The method for preparing the metolazone lyophilized powder preparation according to claim 8, characterized in that: The method comprises the following steps: (1) when the pharmaceutical composition is in powder form, filtering after re-dissolving, or filtering the metolazone inclusion compound solution obtained by "stirring until clear and transparent"; (2) Dry it afterwards; Preferably, the reconstituted solvent is water, more preferably water for injection, the amount of the water for injection is 0.4-2 times the volume of the metolazone inclusion solution, and the volume of the metolazone inclusion solution is the volume of the first solvent as claimed in claim 4 or the total volume of the second solvent and the third solvent; Preferably, the filtration is performed by membrane filtration; Preferably, the drying method is freeze drying, spray drying or reduced pressure drying, more preferably freeze drying; Preferably, before filtering, a freeze-drying protective agent is added for dissolution.

10. Use of a metolazone inclusion compound or a metolazone lyophilized powder preparation in the preparation of a diuretic drug, characterized in that: The metolazone inclusion compound is the metolazone inclusion compound according to claim 1 or 2, or the metolazone inclusion compound prepared by the preparation method according to any one of claims 3 to 6, and the metolazone lyophilized powder preparation is the metolazone lyophilized powder preparation according to claim 8, or the metolazone lyophilized powder preparation obtained by the preparation method according to claim 9.

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