A pharmaceutical composition containing betaine phosphate, a preparation method and application thereof in preparing a drug for treating methylmalonic acidemia combined with homocystinemia

Abstract

The invention discloses a pharmaceutical composition containing phosphobetaine, a preparation method and application of the pharmaceutical composition in preparing a medicine for treating methylmalonic acidemia combined with homocystinemia, belonging to the technical field of betaine. The preparation method comprises the following steps: preparing phosphobetaine and compounding; the compounding comprises the following steps: preparing calcium hydroxycarbonate, compounding once, compounding twice and coating; the pharmaceutical composition containing phosphobetaine of the invention can ensure the therapeutic effect on methylmalonic acidemia combined with homocystinemia, and the preparation method of the pharmaceutical composition containing phosphobetaine of the invention can also improve the purity and yield of phosphobetaine, avoid agglomeration of phosphobetaine crystals, and the prepared pharmaceutical composition containing phosphobetaine has weak moisture absorption, strong fluidity and good high temperature resistance.

Description

Technical Field

[0001] The invention relates to the technical field of betaine, and in particular to a pharmaceutical composition containing betaine phosphate, a preparation method and application of the pharmaceutical composition in preparing a medicine for treating methylmalonic acidemia combined with homocystinemia. Background Art

[0002] Bone diseases are common diseases, including fractures, osteoporosis, arthritis, bone tumors, etc., which manifest as local or systemic bone tissue damage. Bone diseases impair motor function and can also cause a variety of secondary diseases and damage to other systemic organs. Rare diseases and genetic diseases are potentially behind chronic bone diseases, such as different types of homocystinemia.

[0003] Research on the pathophysiological mechanism of chronic bone disease has found that many patients with chronic bone disease suffer from hypophosphatemia. Phosphorus can mediate many biological processes of living organisms. Phosphorus metabolism is essential for maintaining life. Phosphorus metabolism homeostasis depends on the ingenious and complex regulatory mechanism of phosphate between the intestine, kidney, bone, and body fluids. However, compared with the widespread attention paid to calcium and vitamin D, the impact of phosphorus metabolism on chronic bone disease has been neglected, and people's understanding and research on phosphorus metabolism homeostasis lags far behind calcium and vitamin D.

[0004] Therefore, achieving phosphorus metabolism homeostasis by regulating phosphorus metabolism is of great significance for the prevention and treatment of chronic bone disease and its complications.

[0005] Methylmalonic acidemia is a common organic acid metabolic disease. The results of neonatal disease screening and clinical research in my country show that about 70% of methylmalonic acidemia patients have concurrent homocystinemia, that is, methylmalonic acidemia combined with homocystinemia. There are significant individual differences among patients. The disease can occur in all age groups from newborns to the elderly, and it is easy to cause multiple organ and system diseases such as nervous, skeletal, blood, cardiovascular, kidney, and eye. Among them, many patients suffer from osteoporosis and hypophosphatemia.

[0006] Therefore, in the treatment of methylmalonic acidemia combined with homocystinemia, phosphorus metabolism should be regulated to prevent and treat bone age while treating organic acid metabolism.

[0007] Currently, commonly used medicinal phosphorus preparations include alendronate sodium vitamin D3 tablets, vitamin D phosphate calcium gluconate tablets, and fibroblast growth factor 23. However, the aforementioned medicinal phosphorus preparations can only regulate phosphorus metabolism and are not helpful in the treatment of methylmalonic acidemia combined with homocystinemia. In addition, the combined use of multiple drugs increases the cost of treatment and may also cause interactions between drugs, affecting drug efficacy. It may also lead to excessive drug concentrations in the body and cause toxic reactions.

[0008] The inventors have found through experiments that phosphobetaine can regulate homocysteine ​​metabolism and phosphorus metabolism at the same time, and achieve a good therapeutic effect on methylmalonic acidemia combined with homocystinemia. Therefore, the inventors began to study the preparation method of phosphobetaine in order to further study and improve the treatment of methylmalonic acidemia combined with homocystinemia. However, the following problems still exist in the preparation and application:

[0009] First, the inventors react chloroacetic acid with sodium hydroxide to obtain a sodium chloroacetate solution, then react sodium chloroacetate with trimethylamine to obtain a betaine solution, then react betaine with phosphoric acid to obtain a phosphobetaine solution, and finally cool the phosphobetaine solution for crystallization, filter, and dry the filter residue to obtain phosphobetaine. When sodium chloroacetate and trimethylamine are reacted, a large amount of sodium chloride is produced, and the sodium chloride is difficult to completely remove, which affects the purity of the obtained phosphobetaine. In order to solve this problem, the most commonly used method is to increase the concentration of the phosphobetaine solution when cooling and crystallizing the phosphobetaine solution, and wash the filter residue with water for many times after cooling, crystallizing and filtering. However, increasing the concentration of the phosphobetaine solution will cause the crystals of the obtained phosphobetaine to agglomerate, and since phosphobetaine is easily soluble in water, washing the filter residue with water for many times will reduce the yield of phosphobetaine.

[0010] Second, betaine phosphate itself has strong moisture absorption and weak fluidity. In order to solve this problem, the most commonly used method at present is to compound betaine phosphate with calcium salt, specifically, spray-drying betaine phosphate solution and calcium salt solution to obtain a betaine phosphate complex. However, the binding force between betaine phosphate and calcium salt is poor, which leads to easy separation of betaine phosphate and calcium salt during long-term high-temperature storage, thereby reducing the fluidity of the betaine phosphate complex and improving the moisture absorption of the betaine phosphate complex. Summary of the invention

[0011] In view of the deficiencies in the prior art, the present invention provides a pharmaceutical composition containing phosphobetaine, a preparation method and use of the pharmaceutical composition containing phosphobetaine in preparing a medicament for treating methylmalonic acidemia combined with homocystinemia. The pharmaceutical composition containing phosphobetaine of the present invention can ensure the therapeutic effect on methylmalonic acidemia combined with homocystinemia, and the preparation method of the pharmaceutical composition containing phosphobetaine of the present invention can also improve the purity and yield of phosphobetaine and avoid agglomeration of phosphobetaine crystals. In addition, the prepared pharmaceutical composition containing phosphobetaine has weak moisture absorption, strong fluidity and good high temperature resistance.

[0012] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0013] A method for preparing a pharmaceutical composition containing betaine phosphate, comprising the following steps: preparing betaine phosphate, compounding;

[0014] The method for preparing betaine phosphate comprises the following steps: adding a first portion of deionized water and chloroacetic acid into a four-necked flask, controlling the temperature in the four-necked flask to 35-50° C., controlling the stirring speed to 100-150 rpm, stirring until completely dissolved, dripping a sodium hydroxide aqueous solution until the pH value is 4-5, controlling the temperature in the four-necked flask to 20-25° C., slowly introducing trimethylamine, and after the introduction, controlling the temperature in the four-necked flask to 35-55° C., stirring for 5.5-6.5 hours, and obtaining a betaine solution; vacuum concentrating the betaine solution at 70-80° C. to half of the original volume, filtering while hot, taking a filtrate, adding the second portion of deionized water, betaine, and betaine phosphate into a four-necked flask, controlling the stirring speed in the four-necked flask to 100-150 rpm, and stirring at room temperature until After complete dissolution, vacuum concentrate to half of the original volume at 70-80° C., filter while hot, take the filtrate, add it to a four-necked flask, control the stirring speed in the four-necked flask to 100-200 rpm, and drop a phosphoric acid aqueous solution at room temperature until the pH is 2-2.5 to obtain a phosphotinyl solution; divide the phosphotinyl solution into 2 parts, vacuum concentrate one part at 70-80° C. to half of the original volume as a concentrated phosphotinyl solution, add the other part to a beaker, control the temperature in the beaker to 0-5° C., drop the concentrated phosphotinyl solution at a temperature of 30-40° C., let it stand for 18-20 hours after the dropwise addition is completed, filter, take the filter cake, use 1800-2000 mL of deionized water to wash the filter cake, dry and crush the filter cake to obtain phosphotinyl;

[0015] In the preparation of betaine phosphate, the mass ratio of deionized water, chloroacetic acid, trimethylamine, the second portion of deionized water, betaine, and betaine phosphate is 370-400:200-210:128-135:400-450:24-26:20-21;

[0016] The mass concentration of the sodium hydroxide aqueous solution is 15-25%;

[0017] The dropping speed of the sodium hydroxide aqueous solution is 3 to 4 g / min;

[0018] The introduction rate of the trimethylamine is 5-7 g / min;

[0019] The mass concentration of the betaine is 99.5%;

[0020] The mass concentration of the betaine phosphate is 99.5%;

[0021] The mass concentration of the phosphoric acid aqueous solution is 30-40%;

[0022] The drop rate of the concentrated betaine phosphate solution is 2 to 3 g / min;

[0023] The compounding comprises the following steps: preparing calcium hydroxycarbonate, primary compounding, secondary compounding, and coating;

[0024] The method for preparing hydroxy calcium carbonate comprises the following steps: adding calcium chloride, calcium oxide and deionized water into a four-necked flask, controlling the temperature in the four-necked flask to -3°C to -1°C, controlling the stirring speed to 50 to 100 rpm, stirring for 10 to 15 minutes, adding sodium bicarbonate, stirring for 2 to 3 hours, and obtaining a reaction liquid; adding the reaction liquid into a beaker, standing at room temperature for 1.5 to 2 hours, filtering, obtaining a filter cake, drying and crushing the filter cake, and obtaining hydroxy calcium carbonate;

[0025] In the preparation of calcium hydroxycarbonate, the mass ratio of calcium chloride, calcium oxide, deionized water and sodium bicarbonate is 28-30:28.3-30.3:360-380:42.5-45.5;

[0026] The primary composite method comprises adding calcium hydroxycarbonate, betaine phosphate obtained in the step of preparing betaine phosphate, and deionized water into a four-necked flask, controlling the stirring speed of the four-necked flask to 50-100 rpm, stirring at room temperature for 40-50 minutes, and then standing at room temperature for 1.5-2 hours, filtering, taking a filter cake, drying and crushing the filter cake to obtain a primary composite;

[0027] In the first compounding, the mass ratio of calcium hydroxycarbonate, betaine phosphate obtained in the step of preparing betaine phosphate, and deionized water is 48-50:14-15:400-420;

[0028] The secondary compounding method comprises adding the primary compound, sodium stearate and deionized water into a four-necked flask, controlling the temperature in the four-necked flask to 50-60° C., controlling the stirring speed to 50-100 rpm, stirring for 30-40 minutes, filtering, taking the filter cake, drying and crushing the filter cake to obtain the secondary compound;

[0029] In the secondary compounding, the mass ratio of the primary compound, sodium stearate and deionized water is 60-62:15-16:700-750;

[0030] The coating method comprises the following steps: adding the secondary complex and deionized water into a four-necked flask, controlling the stirring speed of the four-necked flask to 50-100 rpm, stirring for 20-30 minutes at room temperature, dripping a calcium chloride solution, stirring for 10-15 minutes after the dripping is completed, filtering, taking a filter cake, washing the filter cake with deionized water for 3-4 times, drying and crushing the filter cake, and obtaining a pharmaceutical composition containing betaine phosphate;

[0031] In the coating, the mass ratio of the secondary composite, deionized water, and calcium chloride solution is 70-72:500-550:35-40;

[0032] The mass concentration of the calcium chloride solution is 10%.

[0033] The dropping speed of the calcium chloride solution is 5-6 g / min.

[0034] A pharmaceutical composition containing betaine phosphate prepared by the above-mentioned preparation method.

[0035] A use of the aforementioned pharmaceutical composition containing phosphobetaine in the preparation of a drug for treating methylmalonic acidemia combined with homocystinemia.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] (1) The preparation method of the pharmaceutical composition containing betaine phosphate of the present invention comprises two steps of preparing betaine phosphate and compounding. In the step of preparing betaine phosphate, the betaine solution is vacuum concentrated and filtered twice, wherein betaine and betaine phosphate are added before the second vacuum concentration. Because the sodium chloride content in the filtrate obtained after the first vacuum concentration and filtration is low and difficult to remove, betaine and betaine phosphate are additionally added in order to further remove the sodium chloride, which not only increases the mass of betaine, but also adds betaine phosphate whose solubility is between that of betaine and sodium chloride, thereby further The remaining sodium chloride is removed in the first step, thereby improving the purity of the obtained phosphotin. In addition, after the phosphotin solution is equally divided, one portion is concentrated as a concentrated phosphotin solution, and the other portion is directly subjected to cooling crystallization. Then, during the cooling crystallization, the concentrated phosphotin solution is heated to 30-40° C. and then dripped into the solution. As the temperature of the concentrated phosphotin solution drops rapidly, a large number of crystal nuclei are generated. After the dripping is completed, the remaining phosphotin solution grows around the crystal nuclei, thereby avoiding the problem of crystal agglomeration when a high-concentration phosphotin solution is used for cooling crystallization, and avoiding the problem of crystal grain agglomeration when a high-concentration phosphotin solution is used for cooling crystallization. When a low-concentration betaine phosphate solution is cooled and crystallized, the crystal nucleus grows too slowly, thereby increasing the yield of the obtained betaine phosphate; in the compounding step, firstly, hydroxy calcium carbonate with hydroxyl groups on the surface is prepared, and then in the first compounding, the hydroxy calcium carbonate is mixed with the betaine phosphate, and the hydroxy calcium carbonate and the betaine phosphate are combined by intermolecular forces to form a primary composite, and then in the second compounding, sodium stearate is mixed with the first composite, and the sodium stearate and the first composite are combined by intermolecular forces to form a secondary composite, and finally the secondary composite is dispersed in water, and calcium chloride is added, and the calcium chloride and the secondary composite are combined. The sodium stearate on the surface reacts to coat a layer of calcium stearate on the surface of the primary composite, thereby obtaining a pharmaceutical composition containing betaine phosphate, wherein the core is calcium carbonate and betaine phosphate and the shell is calcium stearate. There is an intermolecular force between calcium carbonate, betaine phosphate and calcium stearate. Compared with directly mixing calcium carbonate, betaine phosphate and calcium stearate by stirring, the intermolecular force is greater. Therefore, the fluidity and high temperature resistance of the pharmaceutical composition containing betaine phosphate are improved, and the moisture absorption of the pharmaceutical composition containing betaine phosphate is reduced. The composite of calcium carbonate and betaine phosphate can also be more conducive to calcium stearate coating;

[0038] (2) The pharmaceutical composition containing phosphobetaine of the present invention has a good therapeutic effect on methylmalonic acidemia combined with homocystinemia;

[0039] (3) The method for preparing the pharmaceutical composition containing betaine phosphate of the present invention, wherein the mass purity of the prepared betaine phosphate is 99.3-99.4%, and the molar yield is 94.29-96.35%;

[0040] (4) The method for preparing the pharmaceutical composition containing betaine phosphate of the present invention, wherein the prepared betaine phosphate crystals do not have the problem of agglomeration;

[0041] (5) The method for preparing the pharmaceutical composition containing betaine phosphate of the present invention has weak moisture absorption. After the pharmaceutical composition containing betaine phosphate prepared by the present invention is placed in an environment with a temperature of 30°C and a relative humidity of 75°C for 10 days, there is no moisture problem;

[0042] (6) The method for preparing the pharmaceutical composition containing betaine phosphate of the present invention, wherein the prepared pharmaceutical composition containing betaine phosphate has strong fluidity and a repose angle of 27 to 28°;

[0043] (7) The method for preparing the pharmaceutical composition containing betaine phosphate of the present invention has good high temperature resistance. After the pharmaceutical composition containing betaine phosphate prepared by the present invention is sealed and placed in an environment with a temperature of 60°C and a relative humidity of 50°C for 1 day, the angle of repose is 28-29°. DETAILED DESCRIPTION

[0044] In order to more clearly explain the technical features, purposes and effects of the present invention, specific implementation methods of the present invention are now described.

[0045] Example 1

[0046] A method for preparing a pharmaceutical composition containing betaine phosphate, specifically comprising:

[0047] 1. Preparation of betaine phosphate: 370g of deionized water and 200g of chloroacetic acid are added to a four-necked flask, the temperature in the four-necked flask is controlled to 35°C, the stirring speed is controlled to 100rpm, and after stirring until completely dissolved, a sodium hydroxide aqueous solution is added dropwise to a pH of 4, the temperature in the four-necked flask is controlled to 20°C, 128g of trimethylamine is slowly introduced, and after the introduction, the temperature in the four-necked flask is controlled to 35°C, and stirred for 5.5h to obtain a betaine solution; the betaine solution is vacuum concentrated to half of the original volume at 70°C, filtered while hot, and the filtrate is taken, and 400g of deionized water, 24g of betaine with a mass purity of 99.5%, and 20g of betaine phosphate with a mass purity of 99.5% are added to a four-necked flask, the stirring speed in the four-necked flask is controlled to 100rpm, and at room temperature After stirring until completely dissolved, vacuum concentrate to half of the original volume at 70°C, filter while hot, take the filtrate, add it to a four-necked flask, control the stirring speed in the four-necked flask to 100rpm, and drop phosphoric acid aqueous solution at room temperature until the pH is 2 to obtain a phosphotin solution; divide the phosphotin solution into 2 parts, concentrate one part at 70°C in vacuum to half of the original volume as a concentrated phosphotin solution, add the other part to a beaker, control the temperature in the beaker to 0°C, drop the concentrated phosphotin solution at 30°C, let it stand for 18h after the addition is completed, filter, take the filter cake, use 1800mL of deionized water to wash the filter cake, dry and crush the filter cake to obtain 501.46g of phosphotin with a mass purity of 99.3% and a molar yield of 95.77%;

[0048] The mass concentration of the sodium hydroxide aqueous solution is 15%;

[0049] The dropping speed of the sodium hydroxide aqueous solution is 3g / min;

[0050] The feeding speed of the trimethylamine is 5 g / min;

[0051] The mass concentration of the phosphoric acid aqueous solution is 30%;

[0052] The drop rate of the concentrated betaine phosphate solution is 2 g / min;

[0053] The calculation formula of the molar yield of phosphobetaine in this embodiment is: [(mass of the obtained phosphobetaine × mass purity of the obtained phosphobetaine - mass of the added phosphobetaine × 99.5%) / 215.14] / (mass of the added chloroacetic acid / 94.50+mass of the added betaine × 99.5% / 117.15) × 100%;

[0054] 2. Compound:

[0055] (1) Preparation of hydroxy calcium carbonate: 28 g of calcium chloride, 28.3 g of calcium oxide, and 360 g of deionized water were added to a four-necked flask, the temperature in the four-necked flask was controlled to -3°C, the stirring speed was controlled to 50 rpm, and the mixture was stirred for 10 min. 42.5 g of sodium bicarbonate was added and stirred for 2 h to obtain a reaction solution. The reaction solution was added to a beaker, allowed to stand at room temperature for 1.5 h, filtered, and a filter cake was taken. The filter cake was dried and crushed to obtain hydroxy calcium carbonate.

[0056] (2) Primary compounding: 48 g of calcium hydroxycarbonate, 14 g of betaine phosphate obtained in the first step of preparing betaine phosphate, and 400 g of deionized water are added to a four-necked flask, the stirring speed of the four-necked flask is controlled to 50 rpm, and the mixture is stirred at room temperature for 40 min, then allowed to stand at room temperature for 1.5 h, filtered, and the filter cake is taken, dried, and crushed to obtain a primary compound;

[0057] (3) Secondary compounding: add 60 g of the primary compound, 15 g of sodium stearate and 700 g of deionized water into a four-necked flask, control the temperature in the four-necked flask to 50°C, control the stirring speed to 50 rpm, stir for 30 min, filter, take the filter cake, dry and crush the filter cake to obtain a secondary compound;

[0058] (4) Coating: 70 g of the secondary composite and 500 g of deionized water were added to a four-necked flask, the stirring speed of the four-necked flask was controlled to 50 rpm, and the mixture was stirred at room temperature for 20 min. 35 g of calcium chloride solution was added dropwise, and the mixture was stirred for 10 min after the addition was completed. The mixture was filtered, and the filter cake was taken. The filter cake was washed with deionized water for 3 times, and 250 mL of deionized water was used for each washing of the filter residue. The filter cake was dried and crushed to obtain a pharmaceutical composition containing betaine phosphate.

[0059] The mass concentration of the calcium chloride solution is 10%;

[0060] The dropping speed of the calcium chloride solution is 5 g / min.

[0061] This embodiment also provides a pharmaceutical composition containing betaine phosphate prepared by the aforementioned preparation method.

[0062] This embodiment also provides a use of the aforementioned pharmaceutical composition containing betaine phosphate in the preparation of a drug for treating methylmalonic acidemia combined with homocystinemia.

[0063] Example 2

[0064] A method for preparing a pharmaceutical composition containing betaine phosphate, specifically comprising:

[0065] 1. Preparation of betaine phosphate: 380g of deionized water and 205g of chloroacetic acid are added to a four-necked flask, the temperature in the four-necked flask is controlled to 45°C, the stirring speed is controlled to 120rpm, and after stirring until completely dissolved, a sodium hydroxide aqueous solution is added dropwise to a pH of 4.5, the temperature in the four-necked flask is controlled to 22°C, 133g of trimethylamine is slowly introduced, and after the introduction, the temperature in the four-necked flask is controlled to 45°C, and stirred for 6h to obtain a betaine solution; the betaine solution is vacuum concentrated to half of the original volume at 75°C, filtered while hot, and the filtrate is taken, and 420g of deionized water, 25g of betaine with a mass purity of 99.5%, and 20.5g of betaine phosphate with a mass purity of 99.5% are added to a four-necked flask, the stirring speed in the four-necked flask is controlled to 120rpm, and at room temperature After stirring until completely dissolved, vacuum concentrate to half of the original volume at 75°C, filter while hot, take the filtrate, add it to a four-necked flask, control the stirring speed in the four-necked flask to 150rpm, and drop a phosphoric acid aqueous solution at room temperature until the pH is 2.2 to obtain a phosphotinyl solution; divide the phosphotinyl solution into 2 parts, concentrate one part at 75°C to half of the original volume in vacuum as a concentrated phosphotinyl solution, add the other part to a beaker, control the temperature in the beaker to 2°C, drop the concentrated phosphotinyl solution at a temperature of 35°C, let it stand for 19h after the addition is completed, filter, take the filter cake, use 1900mL of deionized water to wash the filter cake, dry and crush the filter cake to obtain 517.21g of phosphotinyl with a mass purity of 99.4% and a molar yield of 96.35%;

[0066] The mass concentration of the sodium hydroxide aqueous solution is 20%;

[0067] The dropping speed of the sodium hydroxide aqueous solution is 3.5 g / min;

[0068] The feeding rate of the trimethylamine is 6 g / min;

[0069] The drop rate of the concentrated betaine phosphate solution is 2.5 g / min;

[0070] The calculation formula for the molar yield of phosphobetaine in this embodiment is the same as the calculation formula for the molar yield of phosphobetaine in Example 1;

[0071] 2. Compound:

[0072] (1) Preparation of hydroxy calcium carbonate: 29 g of calcium chloride, 29.5 g of calcium oxide, and 370 g of deionized water were added to a four-necked flask, the temperature in the four-necked flask was controlled to -2°C, the stirring speed was controlled to 80 rpm, and the mixture was stirred for 12 min. 44 g of sodium bicarbonate was added and stirred for 2.5 h to obtain a reaction solution. The reaction solution was added to a beaker, allowed to stand at room temperature for 2 h, filtered, and a filter cake was taken. The filter cake was dried and crushed to obtain hydroxy calcium carbonate.

[0073] (2) Primary compounding: 49 g of calcium hydroxycarbonate, 14.5 g of betaine phosphate obtained in the first step of preparing betaine phosphate, and 410 g of deionized water were added to a four-necked flask, the stirring speed of the four-necked flask was controlled to 80 rpm, stirred at room temperature for 45 min, and then allowed to stand at room temperature for 2 h, filtered, and the filter cake was taken, and the filter cake was dried and crushed to obtain a primary compound;

[0074] (3) Secondary compounding: 61 g of the primary compound, 15.5 g of sodium stearate and 720 g of deionized water were added into a four-necked flask, the temperature in the four-necked flask was controlled to 55° C., the stirring speed was controlled to 80 rpm, the mixture was stirred for 35 min, and the mixture was filtered to obtain a filter cake, which was then dried and crushed to obtain a secondary compound;

[0075] (4) Coating: 71 g of the secondary composite and 525 g of deionized water were added to a four-necked flask, the stirring speed of the four-necked flask was controlled to 80 rpm, and the mixture was stirred at room temperature for 25 min. 38 g of calcium chloride solution was added dropwise, and the mixture was stirred for 12 min after the addition was completed. The mixture was filtered, and the filter cake was taken. The filter cake was washed with deionized water for 3 times, and 280 mL of deionized water was used for each washing of the filter residue. The filter cake was dried and crushed to obtain a pharmaceutical composition containing betaine phosphate.

[0076] The mass concentration of the calcium chloride solution is 10%.

[0077] The dropping speed of the calcium chloride solution is 5.5 g / min.

[0078] This embodiment also provides a pharmaceutical composition containing betaine phosphate prepared by the aforementioned preparation method.

[0079] This embodiment also provides a use of the aforementioned pharmaceutical composition containing betaine phosphate in the preparation of a drug for treating methylmalonic acidemia combined with homocystinemia.

[0080] Example 3

[0081] A method for preparing a pharmaceutical composition containing betaine phosphate, specifically comprising:

[0082] 1. Preparation of betaine phosphate: 400g of deionized water and 210g of chloroacetic acid are added to a four-necked flask, the temperature in the four-necked flask is controlled to 50°C, the stirring speed is controlled to 150rpm, and after stirring until completely dissolved, a sodium hydroxide aqueous solution is added dropwise to a pH of 5, the temperature in the four-necked flask is controlled to 25°C, 135g of trimethylamine is slowly introduced, and after the introduction, the temperature in the four-necked flask is controlled to 55°C, and stirred for 6.5h to obtain a betaine solution; the betaine solution is vacuum concentrated to half of the original volume at 80°C, filtered while hot, and the filtrate is taken, and 450g of deionized water, 26g of betaine with a mass purity of 99.5%, and 21g of betaine phosphate with a mass purity of 99.5% are added to a four-necked flask, the stirring speed in the four-necked flask is controlled to 150rpm, and stirred at room temperature After stirring until completely dissolved, vacuum concentrate to half of the original volume at 80°C, filter while hot, take the filtrate, add it to a four-necked flask, control the stirring speed in the four-necked flask to 200rpm, and drop phosphoric acid aqueous solution at room temperature until the pH is 2.5 to obtain a phosphotin solution; divide the phosphotin solution into 2 parts, concentrate one part at 80°C in vacuum to half of the original volume as a concentrated phosphotin solution, add the other part to a beaker, control the temperature in the beaker to 5°C, drop the concentrated phosphotin solution at 40°C, let it stand for 20h after the addition is completed, filter, take the filter cake, use 2000mL of deionized water to wash the filter cake, dry and crush the filter cake to obtain 520.11g of phosphotin with a mass purity of 99.3% and a molar yield of 94.29%;

[0083] The mass concentration of the sodium hydroxide aqueous solution is 25%;

[0084] The dropping speed of the sodium hydroxide aqueous solution is 4 g / min;

[0085] The feeding rate of the trimethylamine is 7 g / min;

[0086] The mass concentration of the phosphoric acid aqueous solution is 40%;

[0087] The drop rate of the concentrated betaine phosphate solution is 3 g / min;

[0088] The calculation formula for the molar yield of phosphobetaine in this embodiment is the same as the calculation formula for the molar yield of phosphobetaine in Example 1;

[0089] 2. Compound:

[0090] (1) Preparation of hydroxy calcium carbonate: 30 g of calcium chloride, 30.3 g of calcium oxide, and 380 g of deionized water were added to a four-necked flask, the temperature in the four-necked flask was controlled to -1°C, the stirring speed was controlled to 100 rpm, and the mixture was stirred for 15 min. 45.5 g of sodium bicarbonate was added and stirred for 3 h to obtain a reaction solution. The reaction solution was added to a beaker, allowed to stand at room temperature for 2 h, filtered, and a filter cake was taken. The filter cake was dried and crushed to obtain hydroxy calcium carbonate.

[0091] (2) Primary compounding: 50 g of calcium hydroxycarbonate, 15 g of betaine phosphate obtained in the first step of preparing betaine phosphate, and 420 g of deionized water were added to a four-necked flask, the stirring speed of the four-necked flask was controlled to 100 rpm, stirred at room temperature for 50 min, and then allowed to stand at room temperature for 2 h, filtered, and the filter cake was taken, and the filter cake was dried and crushed to obtain a primary compound;

[0092] (3) Secondary compounding: 62 g of the primary compound, 16 g of sodium stearate and 750 g of deionized water were added to a four-necked flask, the temperature in the four-necked flask was controlled to 60°C, the stirring speed was controlled to 100 rpm, the mixture was stirred for 40 min, filtered, the filter cake was taken, the filter cake was dried and crushed to obtain a secondary compound;

[0093] (4) Coating: 72 g of the secondary composite and 550 g of deionized water were added to a four-necked flask, the stirring speed of the four-necked flask was controlled to 100 rpm, and the mixture was stirred at room temperature for 30 min. 40 g of calcium chloride solution was added dropwise, and the mixture was stirred for 15 min after the addition was completed. The mixture was filtered, and the filter cake was taken. The filter cake was washed with deionized water for 4 times, and 300 mL of deionized water was used for each washing of the filter residue. The filter cake was dried and crushed to obtain a pharmaceutical composition containing betaine phosphate.

[0094] The mass concentration of the calcium chloride solution is 10%.

[0095] The dropping speed of the calcium chloride solution is 6 g / min.

[0096] This embodiment also provides a pharmaceutical composition containing betaine phosphate prepared by the aforementioned preparation method.

[0097] This embodiment also provides a use of the aforementioned pharmaceutical composition containing betaine phosphate in the preparation of a drug for treating methylmalonic acidemia combined with homocystinemia.

[0098] Comparative Example 1

[0099] Adjustments were made based on the method for preparing the pharmaceutical composition containing betaine phosphate in Example 2, and the specific adjustments were as follows:

[0100] In the first step of preparing betaine phosphate, the addition of betaine with a mass purity of 99.5% and betaine phosphate with a mass purity of 99.5% are omitted, and the process of dividing the betaine phosphate solution into two parts and the subsequent dropwise addition process are omitted. Specifically, the first step of preparing betaine phosphate is changed to:

[0101] 380g of deionized water and 205g of chloroacetic acid were added to a four-necked flask, the temperature in the four-necked flask was controlled to 45°C, the stirring speed was controlled to 120rpm, and after stirring until completely dissolved, sodium hydroxide aqueous solution was added dropwise until the pH value was 4.5, the temperature in the four-necked flask was controlled to 22°C, 131g of trimethylamine was slowly introduced, and after the introduction, the temperature in the four-necked flask was controlled to 45°C, and stirred for 6h to obtain a betaine solution; the betaine solution was vacuum concentrated to half of the original volume at 75°C, and the mixture was stirred for 6h. Hot filtration, taking the filtrate, adding it to a four-necked flask, controlling the stirring speed in the four-necked flask to 150 rpm, adding phosphoric acid aqueous solution at room temperature until the pH is 2.2, and obtaining a phosphobetaine solution; adding the phosphobetaine solution to a beaker, controlling the temperature in the beaker to 2°C, standing for 19 hours, filtering, taking the filter cake, washing the filter cake with 1900 mL of deionized water, drying and crushing the filter cake, and obtaining 447.38 g of phosphobetaine with a mass purity of 97.2% and a molar yield of 93.18%;

[0102] The mass concentration of the sodium hydroxide aqueous solution is 20%;

[0103] The dropping speed of the sodium hydroxide aqueous solution is 3.5 g / min;

[0104] The feeding rate of the trimethylamine is 6 g / min;

[0105] The mass concentration of the phosphoric acid aqueous solution is 35%.

[0106] The calculation formula of the molar yield of phosphobetaine in this comparative example is: (mass of the obtained phosphobetaine × mass purity of the obtained phosphobetaine / 215.14) / (mass of the added chloroacetic acid / 94.50)×100%;

[0107] The rest of the technical solutions are the same as those in Example 2.

[0108] This comparative example also provides a pharmaceutical composition containing betaine phosphate prepared by the aforementioned preparation method.

[0109] This comparative example also provides an application of the aforementioned pharmaceutical composition containing betaine phosphate in the preparation of a drug for treating methylmalonic acidemia combined with homocystinemia.

[0110] Comparative Example 2

[0111] Adjustments were made based on the method for preparing the pharmaceutical composition containing betaine phosphate in Example 2, and the specific adjustments were as follows:

[0112] The second step of compounding is changed to directly mixing calcium carbonate, betaine phosphate and calcium stearate, specifically: 49g of calcium carbonate, 14.5g of betaine phosphate obtained in the step of preparing betaine phosphate in step 1, and 15.5g of calcium stearate are added to a beaker, and stirred at room temperature at a stirring speed of 80 rpm for 45 minutes to obtain a pharmaceutical composition containing betaine phosphate;

[0113] The rest of the technical solutions are the same as those in Example 2.

[0114] This comparative example also provides a pharmaceutical composition containing betaine phosphate prepared by the aforementioned preparation method.

[0115] This comparative example also provides an application of the aforementioned pharmaceutical composition containing betaine phosphate in the preparation of a drug for treating methylmalonic acidemia combined with homocystinemia.

[0116] Test Example 1

[0117] It was observed whether the crystals of phosphobetaine obtained in the step of preparing phosphobetaine in step 1 in Example 1-3 and Comparative Example 1-2 had agglomeration problems, and the observation results were as follows:

[0118]

[0119] Test Example 2

[0120] 50 g of the pharmaceutical composition containing betaine phosphate obtained in the second compounding step in Example 1-3 and Comparative Example 1-2 were respectively placed in a 500 mL beaker with an open mouth, and then placed in an environment with a temperature of 30° C. and a relative humidity of 75° C. for 10 days to observe whether there was a moisture problem. The observation results are as follows:

[0121]

[0122] Test Example 3

[0123] The angle of repose of the pharmaceutical composition containing betaine phosphate obtained in the second compounding step in Example 1-3 and Comparative Example 1-2 was tested, and the test results are as follows:

[0124]

[0125] Then, the pharmaceutical composition containing betaine phosphate obtained in the second compounding step in Example 1-3 and Comparative Example 1-2 was sealed in a packaging bag, and then placed in an environment with a temperature of 60° C. and a relative humidity of 50° C. for 1 day, and then the angle of repose was tested. The test results are as follows:

[0126]

[0127] Test Example 4

[0128] The pharmaceutical composition containing betaine phosphate obtained in the second compounding step in Example 2 was mixed with starch and sodium carboxymethyl starch in a mass ratio of 80:15:5 and then tableted to prepare betaine phosphate tablets with a specification of 300 mg. The tablets were then tested for efficacy. The specific cases are as follows:

[0129] Case 1:

[0130] Male, 13 years old, healthy before 12 years old, began to grow rapidly at 12 years old, voice change, leg pain, irritability, difficulty falling asleep, and decreased academic performance. The local hospital suspected "growing pains, adolescent mood disorders" and gave him vitamin D and calcium carbonate D3 orally. Although the leg pain improved after two weeks, he refused to go to school, talked nonsense, had hallucinations and anorexia. The psychiatric department considered "schizophrenia" and recommended further examination. Blood tests found macrocytic anemia, significantly increased homocysteine ​​in the blood (160μmol / L), decreased 25-hydroxyvitamin D, and significantly increased methylmalonic acid in the urine. Magnetic resonance imaging showed mild cerebral atrophy. Genetic analysis found MMACHC double heterozygous mutations, and was diagnosed with methylmalonic acidemia combined with homocystinemia (cblC type). Hydroxocobalamin was given intramuscular injection (10 mg per day), oral phosphobetaine tablets (15 tablets per day), and vitamin AD drop pills (1 tablet per day). A follow-up examination one week later showed that the anemia had improved and the blood homocysteine ​​level had dropped significantly (57μmol / L). After half a year of treatment, the child returned to school and maintained treatment. He is now 15 years old and leads a normal life without any mental abnormalities or leg pain.

[0131] Case 2:

[0132] Female, came to the hospital at the age of 7 and a half. The child had myopia since childhood, strabismus was discovered at the age of 3, "lens dislocation" was diagnosed at the age of 6, and received "binocular intraocular lens replacement". At the age of 7 years and 3 months, she had headaches and vomiting, and then paralysis of the left limbs. The homocysteine ​​in her serum was significantly increased (209μmol / L), and the methionine was significantly increased (851μmol / L). Vitamin D was decreased. Brain magnetic resonance imaging showed right frontal lobe cerebral infarction, which was consistent with venous sinus thrombosis. Chest CT showed pulmonary embolism, abnormal electroencephalogram, and whole exome analysis detected compound heterozygous mutations in the CBS gene. She was diagnosed with homocystinemia type 1 caused by cystathionine β-synthase deficiency. She was given large doses of vitamin B6 (500 mg per day), oral phosphobetaine tablets (30 tablets per day), and a low-methionine diet. The child is now 9 years old and her condition is stable. Both blood homocysteine ​​and methionine have decreased significantly. She can attend school normally and has good growth and development.

[0133] Case 3:

[0134] Male, first diagnosed at the age of 3, with physical growth retardation, short stature, multiple fractures, motor backwardness, walking at the age of 2, unstable gait, frequent urination, frequent leg and limb pain, sleep disorders, crying and restlessness, vitamin D and calcium supplementation were ineffective. Examination found that blood vitamin D and blood calcium were normal, blood phosphorus was significantly reduced (0.85mmol / L), serum total homocysteine ​​was increased, glomerular function was normal, amino acids in urine were increased, urine calcium / creatinine ratio was significantly increased (1.2), trace albumin in urine was increased, X-ray examination found multiple fractures in the limbs, indicating renal tubular damage, renal osteodystrophy, hypophosphatemia, mild homocystinemia, oral administration of betaine phosphate tablets (6 tablets per day), vitamin B6 and niacin, the child's condition gradually improved, blood phosphorus increased, physical growth rate accelerated, and intelligence and motor skills improved significantly, no fractures occurred, now 5 years old, can go to kindergarten normally.

[0135] Unless otherwise specified, all percentages used in the present invention are by mass.

[0136] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a pharmaceutical composition containing betaine phosphate, characterized in that: The method comprises the following steps: preparing betaine phosphate, compounding; The method for preparing betaine phosphate comprises the following steps: mixing a first portion of deionized water and chloroacetic acid, stirring at 35-50° C. until completely dissolved, dripping a sodium hydroxide aqueous solution until the pH value is 4-5, slowly introducing trimethylamine at 20-25° C., and stirring at 35-55° C. for 5.5-6.5 hours after the introduction to obtain a betaine solution; vacuum concentrating the betaine solution at 70-80° C. to half of the original volume, filtering while hot, taking a filtrate, mixing the filtrate with a second portion of deionized water, betaine, and betaine phosphate, stirring at room temperature until completely dissolved, and evaporating at 70-80° C. The phosphate betaine solution is divided into two equal parts, one of which is concentrated to half of the original volume under vacuum at 70-80° C. as a concentrated phosphate betaine solution, and the other part is cooled to 0-5° C., and the concentrated phosphate betaine solution at a temperature of 30-40° C. is added dropwise, and after the addition is completed, the mixture is allowed to stand for 18-20 hours, filtered, and a filter cake is taken. After the filter cake is washed, the filter cake is dried and crushed to obtain phosphate betaine; The compounding comprises the following steps: preparing calcium hydroxycarbonate, primary compounding, secondary compounding, and coating; The method for preparing hydroxy calcium carbonate comprises: mixing calcium chloride, calcium oxide and deionized water, stirring at -3°C to -1°C for 10 to 15 minutes, adding sodium bicarbonate, stirring for 2 to 3 hours to obtain a reaction solution; allowing the reaction solution to stand at room temperature for 1.5 to 2 hours, filtering, taking a filter cake, drying and crushing the filter cake to obtain hydroxy calcium carbonate; The primary composite method comprises mixing calcium hydroxycarbonate, betaine phosphate obtained in the step of preparing betaine phosphate, and deionized water, stirring at room temperature for 40 to 50 minutes, and then standing at room temperature for 1.5 to 2 hours, filtering, taking a filter cake, drying and crushing the filter cake to obtain a primary composite; The secondary compounding method comprises mixing the primary compound, sodium stearate and deionized water, stirring at 50-60° C. for 30-40 minutes, filtering, taking the filter cake, drying and crushing the filter cake to obtain the secondary compound; The coating method comprises the following steps: mixing the secondary complex and deionized water, stirring for 20 to 30 minutes at room temperature, dropping a calcium chloride solution, stirring for 10 to 15 minutes after the dropping is completed, filtering, taking a filter cake, washing the filter cake, drying and crushing the filter cake to obtain a pharmaceutical composition containing betaine phosphate.

2. The method for preparing the pharmaceutical composition containing betaine phosphate according to claim 1, characterized in that: In the preparation of betaine phosphate, the mass ratio of the first portion of deionized water, chloroacetic acid, trimethylamine, the second portion of deionized water, betaine, and betaine phosphate is 370-400:200-210:128-135:400-450:24-26:20-21.

3. The method for preparing the pharmaceutical composition containing betaine phosphate according to claim 1, characterized in that: In the preparation of betaine phosphate, the mass concentration of the sodium hydroxide aqueous solution is 15-25%; The dropping speed of the sodium hydroxide aqueous solution is 3 to 4 g / min; The introduction rate of the trimethylamine is 5-7 g / min; The mass concentration of the betaine is 99.5%; The mass concentration of the betaine phosphate mixed with the filtrate is 99.5%; The mass concentration of the phosphoric acid aqueous solution is 30-40%; The dropwise addition rate of the concentrated betaine phosphate solution is 2 to 3 g / min.

4. The method for preparing the pharmaceutical composition containing betaine phosphate according to claim 1, characterized in that: In the preparation of calcium hydroxycarbonate, the mass ratio of calcium chloride, calcium oxide, deionized water and sodium bicarbonate is 28-30:28.3-30.3:360-380:42.5-45.

5.

5. The method for preparing the pharmaceutical composition containing betaine phosphate according to claim 1, characterized in that: In the primary compounding, the mass ratio of calcium hydroxycarbonate, betaine phosphate obtained in the betaine phosphate preparation step, and deionized water is 48-50:14-15:400-420.

6. The method for preparing the pharmaceutical composition containing betaine phosphate according to claim 1, characterized in that: In the secondary compounding, the mass ratio of the primary compound, sodium stearate and deionized water is 60-62:15-16:700-750.

7. The method for preparing the pharmaceutical composition containing betaine phosphate according to claim 1, characterized in that: In the coating, the mass ratio of the secondary composite, deionized water, and calcium chloride solution is 70-72:500-550:35-40; The mass concentration of the calcium chloride solution is 10%; The dropping speed of the calcium chloride solution is 5-6 g / min.

8. A pharmaceutical composition containing betaine phosphate prepared by the preparation method according to any one of claims 1 to 7.

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