Deacetylated lanolin freeze-dried powder and preparation process thereof

Abstract

The present invention belongs to the technical field of pharmaceutical preparations for treating cardiovascular diseases, and specifically relates to a deacetylated lanthanoside freeze-dried powder and a preparation process thereof. The deacetylated lanthanoside freeze-dried powder of the present invention is composed of 0.2 to 0.6 parts by weight of deacetylated lanthanoside, 8 to 15 parts by weight of mannitol, 0.5 to 1.5 parts by weight of ethyl maltol, 2 to 5 parts by weight of calcium gluconate, and 1 to 3 parts by weight of phospholipids. By optimizing the prescription and the preparation process, a deacetylated lanthanoside freeze-dried powder with high stability is provided, which overcomes the defect of deacetylated lanthanoside being hygroscopic, reduces the occurrence of adverse reactions, and improves the medication safety of the deacetylated lanthanoside preparation.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations for treating cardiovascular diseases, and particularly relates to deacetyl lanatoside freeze-dried powder and a preparation process thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Deacetylated lanugin is a compound extracted from lanugin, which has multiple pharmacological activities and medical application values. The chemical formula of deacetylated lanugin is C 47 H 74 O 19 , white crystalline powder, odorless, bitter, hygroscopic, slightly soluble in methanol, very slightly soluble in ethanol, almost insoluble in water or chloroform. It is mainly used as a cardiotonic in clinical practice for acute and chronic systolic heart failure, atrial fibrillation and paroxysmal supraventricular tachycardia. It can inhibit Na-K-ATPase on the myocardial cell membrane, reduce sodium-potassium exchange, increase intracellular sodium ions, sodium-calcium exchange, increase intracellular calcium ions, act on contractile proteins, and exert positive inotropic effects. Deacetyl lanthrin is a fast cardiac glycoside that can enhance myocardial contractility, slow heart rate and conduction, and has little effect on coronary artery contraction and cardiac conduction system. It has a good therapeutic effect on senile heart failure and is effective quickly.

[0004] Li Geyuan et al. Study on impurities of deacetyl scutellarin injection [J]. Pharmacy and Clinical Research, 2011, 19(6): 508-510. It was reported that impurities and / or related substances in deacetyl scutellarin injection can cause adverse reactions such as arrhythmia, gastrointestinal discomfort, nausea, vomiting, etc., thus affecting the safety of deacetyl scutellarin injection.

[0005] Chinese patent publication number CN102961326A discloses a method for preparing deacetylated scutellarin injection, which specifically comprises: reducing the temperature of water for injection to 35-42°C; adopting heat treatment at 100°C for 30 min or not performing heat treatment, and replacing ethanol (re-distilled) with ethanol (medicinal) for direct use without re-distillation, thereby saving the time and cost of re-distillation and ensuring quality.

[0006] Chinese patent publication number CN116440071A discloses a deacetylated scutellarin injection and a preparation method thereof, wherein the pH value of the deacetylated scutellarin aqueous solution is adjusted to 6.4-6.9 with a weakly alkaline substance, and the degradation of the deacetylated scutellarin in the liquid preparation under high temperature conditions is inhibited by controlling the pH, thereby reducing the generation of impurities such as impurities C and impurity D.

[0007] Chinese patent publication number CN112156069A discloses a production process for deacetylated scutellarin injection, comprising the following steps: encapsulating a mixture containing 0.2 mg / ml deacetylated scutellarin, 0.076 g / ml ethanol, 0.149 g / ml glycerol and water for injection, nitrogen filling before and after encapsulation, and sampling and testing after placing at high temperature (60±2°C) for 10 days. The results show that the product quality using the double nitrogen filling process is stable and the impurity growth is small. Summary of the invention

[0008] In order to overcome the deficiencies of the prior art, the present invention provides a highly stable deacetylated lanthrin freeze-dried powder by optimizing the prescription and preparation process, thereby overcoming the defect of deacetylated lanthrin being hygroscopic and improving the medication safety of deacetylated lanthrin preparations.

[0009] The first object of the present invention is to provide a formula of deacetylated lanthanoside freeze-dried powder, which consists of 0.2-0.6 parts by weight of deacetylated lanthanoside, 8-15 parts by weight of mannitol, 0.5-1.5 parts by weight of ethyl maltol, 2-5 parts by weight of calcium gluconate, and 1-3 parts by weight of phospholipids.

[0010] Since deacetyl lanthrin is poorly soluble in water, phospholipids are added to aid dissolution, and the phospholipids are selected from at least one of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol, and phosphatidylserine.

[0011] The second object of the present invention is to provide a method for preparing the above-mentioned deacetylated lanolin freeze-dried powder, the method comprising:

[0012] (1) Mixing deacetylated lanolin and phospholipids, adding an appropriate amount of solvent to dilute, and obtaining an intermediate solution for later use;

[0013] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0014] (3) The mixture is freeze-dried, pre-frozen at 0°C to 10°C at normal pressure for 1 to 3 hours, ethyl maltol is added, mixed evenly, sterile filtered (0.22 μm), the temperature is controlled at -40°C to -50°C at normal pressure for 3 to 5 hours, the pressure is reduced in vacuum and the temperature is controlled at -20°C to -30°C for 2 to 4 hours, the temperature is reduced in vacuum to -35°C to -45°C for 1 to 3 hours, the temperature is raised to -5°C to 5°C for 3 to 5 hours, and the mixture is sealed.

[0015] In order to improve the compatibility of deacetylated lanolin with phospholipids, the solvent is selected from at least one of ethanol, water and methanol, and is preferably an ethanol aqueous solution with a mass fraction of 30-40%.

[0016] Compared with the prior art, the technical effects of the present invention are:

[0017] (1) The present invention adds a certain amount of ethyl maltol to the prescription, and adds the material after pre-freezing. Experimental data show that this method effectively overcomes the defect that deacetylated scutellarin is easy to absorb moisture. In long-term tests, the deacetylated scutellarin freeze-dried powder of the present invention has low moisture absorption and weight gain and high stability.

[0018] (2) The present invention optimizes the prescription and preparation process, and inhibits the production of deacetylated lanolin-related substances under the coordination of multiple factors, thereby reducing the occurrence of adverse reactions and improving the safety of medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 : Moisture absorption and weight gain of deacetyl lanolin freeze-dried powder of groups A to I.

[0020] Figure 2 : Changes in the contents of related substances in lyophilized deacetylated lanolin powders of Examples 1 to 5 and Comparative Examples 1 to 5. DETAILED DESCRIPTION

[0021] In order to make the purpose and technical solution of the present invention clearer, the present invention is further described below in conjunction with embodiments, but the protection scope of the present invention is not limited to these embodiments, and the embodiments are only used to explain the present invention. It should be understood by those skilled in the art that any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the protection scope of the present invention.

[0022] Study on the dosage of ethyl maltol and its moisture-proof effect

[0023] In order to explore the moisture-proof effect of the dosage of ethyl maltol on the deacetylated serotonin freeze-dried powder, the following groups were specially designed. Each group of deacetylated serotonin freeze-dried powder was placed in a storage condition of 40±2℃ and 75%±5% for 90 days, and the moisture absorption and weight gain of each group of deacetylated serotonin freeze-dried powder was tested.

[0024] Table 1A~I group deacetyl lanolin freeze-dried powder formula

[0025]

[0026] Preparation method of group A to I deacetyl lanolin freeze-dried powder:

[0027] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 35% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0028] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0029] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, ethyl maltol is added, mixed evenly, and sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced and the temperature is controlled at -25°C under vacuum for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and the mixture is sealed.

[0030] Deacetylated scutellaria baicalensis is hygroscopic and will absorb moisture even if it is made into freeze-dried powder. Figure 1 The results are as follows: The moisture absorption and weight gain of the lyophilized powder of deacetylated lanthanoside in groups A to I. The experiment shows that the lyophilized powder of deacetylated lanthanoside of the present invention preferably has a certain amount of ethyl maltol added as an auxiliary material, which can effectively prevent the deacetylated lanthanoside from deliquescence.

[0031] Example 1 Lyophilized powder of acetylated lanolin

[0032] formula:

[0033]

[0034]

[0035] Preparation method:

[0036] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 35% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0037] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0038] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, ethyl maltol is added, mixed evenly, and sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced and the temperature is controlled at -25°C under vacuum for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and the mixture is sealed.

[0039] Example 2 Lyophilized powder of acetylated lanolin

[0040] formula:

[0041]

[0042] Preparation method:

[0043] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 30% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0044] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0045] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, ethyl maltol is added, mixed evenly, and sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced and the temperature is controlled at -25°C under vacuum for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and the mixture is sealed.

[0046] Example 3 Acetyl lanolin freeze-dried powder

[0047] formula:

[0048]

[0049]

[0050] Preparation method:

[0051] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 40% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0052] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0053] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, ethyl maltol is added, mixed evenly, and sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced and the temperature is controlled at -25°C under vacuum for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and the mixture is sealed.

[0054] Example 4 Acetyl lanolin freeze-dried powder

[0055] formula:

[0056]

[0057] Preparation method:

[0058] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 30% by mass methanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0059] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0060] (3) The mixture is freeze-dried, pre-frozen at 0°C for 1 hour, ethyl maltol is added, mixed evenly, sterile filtered (0.22 μm filter membrane), the temperature is controlled at -40°C at normal pressure for 3 hours, the pressure is reduced and the temperature is controlled at -20°C for 2 hours, the temperature is reduced to -35°C under vacuum for 1 hour, the temperature is raised to -5°C under vacuum for 3 to 5 hours, and sealed.

[0061] Example 5 Acetyl lanolin freeze-dried powder

[0062] formula:

[0063]

[0064]

[0065] Preparation method:

[0066] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 40% methanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0067] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0068] (3) The mixture is freeze-dried, pre-frozen at 10°C at normal pressure for 3 hours, ethyl maltol is added, mixed evenly, sterile filtered, the temperature is controlled at -50°C at normal pressure for 5 hours, the pressure is reduced and the temperature is controlled at -30°C under vacuum for 4 hours, the temperature is lowered to -45°C under vacuum for 3 hours, the temperature is raised to 5°C under vacuum for 5 hours, and the mixture is sealed.

[0069] Comparative Example 1: Lyophilized powder of acetylated lanolin

[0070] formula:

[0071]

[0072] Preparation method:

[0073] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 35% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0074] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0075] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced in vacuum and the temperature is controlled at -25°C for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and sealed.

[0076] Comparative Example 2: Lyophilized powder of acetylated lanolin

[0077] formula:

[0078]

[0079]

[0080] Preparation method:

[0081] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 35% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0082] (2) dissolving mannitol, calcium gluconate and ethyl maltol in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly to obtain a mixture for later use;

[0083] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced in vacuum and the temperature is controlled at -25°C for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and sealed.

[0084] Comparative Example 3: Lyophilized powder of acetylated lanolin

[0085] formula:

[0086]

[0087] Preparation method:

[0088] (1) dissolving deacetylated lansanthin in an appropriate amount of 35% by mass ethanol aqueous solution to obtain an intermediate solution for later use;

[0089] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0090] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, ethyl maltol is added, mixed evenly, and sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced and the temperature is controlled at -25°C under vacuum for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and the mixture is sealed.

[0091] Comparative Example 4: Lyophilized powder of acetylated lanolin

[0092] formula:

[0093]

[0094] Preparation method:

[0095] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 35% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0096] (2) dissolving mannitol in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0097] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, ethyl maltol is added, mixed evenly, and sterile filtered (0.22 μm filter membrane), the temperature is controlled at -45°C at normal pressure for 4 h, the pressure is reduced and the temperature is controlled at -25°C under vacuum for 3 h, the temperature is lowered to -40°C under vacuum for 2 h, the temperature is raised to 0°C under vacuum for 4 h, and the mixture is sealed.

[0098] Comparative Example 5: Lyophilized powder of acetylated lanolin

[0099] formula:

[0100]

[0101] Preparation method:

[0102] (1) Mix deacetylated lanolin and phospholipids, add an appropriate amount of 35% ethanol aqueous solution to dilute, and obtain an intermediate solution for later use;

[0103] (2) dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for later use;

[0104] (3) The mixture is freeze-dried, pre-frozen at 5°C at normal pressure for 2 h, ethyl maltol is added, mixed evenly, sterile filtered (0.22 μm filter membrane), vacuum-decompressed and temperature controlled at -40°C for 3 h, vacuum-controlled temperature is lowered to -15°C for 2 h, vacuum-controlled temperature is raised to 10°C for 4 h, and sealed.

[0105] Stability of Deacetylated Lanatin Freeze-dried Powders of Examples 1 to 5 and Comparative Examples 1 to 5

[0106] Related substances

[0107] The related substances of deacetylated lanolin are determined according to the high performance liquid chromatography method (General Rule 0512) of the Chinese Pharmacopoeia. Test solution: Take this product, add a small amount of methanol to dissolve it by ultrasound, and dilute it with the mobile phase to make a solution containing about 0.2 mg per 1 ml. Control solution: Accurately measure 1 ml of the test solution, place it in a 100 ml volumetric flask, dilute it to the scale with the mobile phase, and shake it well. Chromatographic conditions: Use octadecylsilane bonded silica gel as the filler; use water as mobile phase A, acetonitrile-methanol (22:14) as mobile phase B, and perform gradient elution according to Table 2; the detection wavelength is 220 nm; the injection volume is 20 μl. System suitability requirements: The separation degree between the deacetylated lanolin peak and the adjacent impurity peak should meet the requirements. Determination method: Accurately measure the test solution and the control solution, inject them into the liquid chromatograph respectively, and record the chromatogram. Limit: If there are impurity peaks in the chromatogram of the test solution, the sum of the areas of the impurity peaks shall not be greater than 5 times (5.0%) the area of ​​the main peak of the control solution.

[0108] Table 2 Gradient elution program

[0109]

[0110]

[0111] The deacetylated lanolin freeze-dried powders of Examples 1 to 5 and Comparative Examples 1 to 5 were placed at 25° C. and 60% relative humidity for 12 months for a long-term test. The contents of related substances were detected on the first day, at the end of the second month, at the end of the sixth month, and at the end of the twelfth month during the experiment.

[0112] Figure 2 The following is a graph showing the changes in the contents of related substances in the lyophilized powder of deacetylated lanolin of Examples 1 to 5 and Comparative Examples 1 to 5. The experimental data show that the lyophilized powder of deacetylated lanolin of the present invention can remain stable in long-term tests, and the contents of related substances increase slowly or remain essentially unchanged.

[0113] Moisture absorption and weight gain

[0114] The deacetylated lanolin freeze-dried powders of Examples 1 to 5 and Comparative Examples 1 to 5 were placed at 25° C. and 60% relative humidity for 12 months for a long-term test. The powders were weighed at the end of the 12th month of the test period to calculate the weight gain due to moisture absorption.

[0115] Table 3 Hygroscopic weight gain of acetylanthin freeze-dried powder of Examples 1 to 5 and Comparative Examples 1 to 5

[0116] Hygroscopic weight gain (%) Example 1 0.11 Example 2 0.12 Example 3 0.10 Example 4 0.17 Example 5 0.14 Comparative Example 1 0.83 Comparative Example 2 0.77 Comparative Example 3 0.21 Comparative Example 4 0.24 Comparative Example 5 0.37

[0117] Experiments show that the present invention overcomes the defect that deacetylated lanthrin is hygroscopic. The deacetylated lanthrin freeze-dried powders of Examples 1 to 5 have low moisture absorption and weight gain in long-term tests and high stability.

Claims

1. A deacetylated lanolin freeze-dried powder, characterized in that: The deacetylated lanthanoside freeze-dried powder is composed of 0.2 to 0.6 parts by weight of deacetylated lanthanoside, 8 to 15 parts by weight of mannitol, 0.5 to 1.5 parts by weight of ethyl maltol, 2 to 5 parts by weight of calcium gluconate, and 1 to 3 parts by weight of phospholipids; The preparation method of the deacetylated lanolin freeze-dried powder is as follows: Step (1): mixing deacetylated lansanthin with phospholipids, adding a suitable amount of solvent to dilute, and obtaining an intermediate solution for standby use; Step (2): dissolving mannitol and calcium gluconate in an appropriate amount of water, adding the intermediate solution of step (1), mixing evenly, and obtaining a mixture for standby use; Step (3): freeze-dry the mixture, pre-freeze at 0°C to 10°C at normal pressure for 1 to 3 hours, add ethyl maltol, mix well, filter aseptically, control the temperature at -40°C to -50°C at normal pressure for 3 to 5 hours, reduce the pressure under vacuum and control the temperature at -20°C to -30°C for 2 to 4 hours, reduce the temperature under vacuum to -35°C to -45°C for 1 to 3 hours, increase the temperature under vacuum to -5°C to 5°C for 3 to 5 hours, seal, and obtain.

2. The deacetylated lanolin freeze-dried powder according to claim 1, characterized in that The phospholipid is selected from at least one of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol and phosphatidylserine.

3. The freeze-dried powder of deacetylated lanatoside according to claim 1, characterized in that The deacetyl lanthrin freeze-dried powder is composed of 0.4 parts by weight of deacetyl lanthrin, 11 parts by weight of mannitol, 0.9 parts by weight of ethyl maltol, 3.5 parts by weight of calcium gluconate, and 2.2 parts by weight of phospholipids.

4. The deacetylated lanolin freeze-dried powder according to claim 1 or 2, characterized in that: The phospholipid is phosphatidylcholine and / or phosphatidylethanolamine.

5. The freeze-dried powder of deacetylated lanolin according to claim 1 or 2, characterized in that: The phospholipid is phosphatidylethanolamine.

6. The deacetylated lanatoside freeze-dried powder according to claim 1, characterized in that: The solvent is selected from at least one of ethanol, water and methanol.

7. The freeze-dried powder of deacetylated lanolin according to claim 1, characterized in that The solvent is an ethanol aqueous solution with a mass fraction of 30-40%.

8. The freeze-dried powder of deacetylated lanatoside according to claim 1, characterized in that The step (3) is as follows: freeze-drying the mixture, pre-freezing at 5°C at normal pressure for 2 hours, adding ethyl maltol, mixing evenly, aseptically filtering, controlling the temperature at -45°C at normal pressure for 4 hours, reducing the pressure under vacuum and controlling the temperature at -25°C for 3 hours, lowering the temperature under vacuum to -40°C for 2 hours, raising the temperature under vacuum to 0°C for 4 hours, and sealing to obtain the product.

9. The freeze-dried powder of deacetylated lanatoside according to claim 1, characterized in that The filtration was performed using a 0.22 μm filter membrane.

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