Toremifene citrate liposome, preparation method and preparation

Abstract

The present invention belongs to the technical field of pharmaceutical preparations and specifically relates to a toremifene citrate liposome, a preparation method, and a preparation. The present invention encapsulates toremifene citrate in a lipid carrier composed of toremifene citrate, peanut lecithin, cholesterol, and calcium propionate. The preparation method is optimized to provide a toremifene citrate liposome with high drug loading, high encapsulation efficiency, and high stability. The liposome is then further prepared with pharmaceutically acceptable excipients into a toremifene citrate preparation, which improves oral bioavailability, reduces impurity content, and improves the stability of the preparation.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a toremifene citrate liposome, a preparation method and a preparation. Background Art

[0002] Toremifene citrate was first marketed in Finland in 1988. Its chemical name is 2[4-(4-chloro-1,2-diphenyl-1-butenyl)phenoxy-N,N-dimethylethylamine citrate, and its molecular formula is C 32 H 36 ClNO8, molecular weight 598.08. Toremifene citrate is a nonsteroidal triphenylethylene derivative that binds to estrogen receptors and can produce estrogen-like or anti-estrogen effects, or both. It can be used as an oral selective estrogen receptor modulator for the treatment of advanced (metastatic) breast cancer.

[0003] Toremifene citrate is a white or off-white crystalline powder, odorless, slightly soluble in methanol or ethanol, very slightly soluble in acetone, almost insoluble in water, and readily soluble in glacial acetic acid. Toremifene citrate's poor water solubility results in low oral bioavailability, limiting its widespread clinical application.

[0004] According to the literature, Wang Zhuo reported the dissolution characteristics of toremifene citrate tablets in different media in the Chinese Journal of Pharmaceutical Industry. Toremifene citrate was almost insoluble in pH 6.8 phosphate buffer. The dissolution percentages in 0.1 mol / L HCl, solutions containing 0.1%, 0.5%, and 1.0% sodium lauryl sulfate, and 10% ethanol after 60 min were (44.26±13.00)%, (27.72±12.58)%, (46.20±9.68)%, (87.15±1.16)%, and (78.27±9.38)%, respectively. As is well known, gastric juice is acidic, and the dissolution of toremifene citrate in 0.1 mol / L HCl is slow and unstable, with a dissolution percentage of only 44.3% after 60 min, indicating low oral bioavailability. (Wang Zhuo, Zhang Limei, Yang Zhangwei, et al. Dissolution characteristics of toremifene citrate tablets in different media [J]. Chinese Journal of Pharmaceutical Industry, 2004 (6).)

[0005] Liposomes are ultramicrospherical carrier formulations formed by a lipid bilayer. They are a typical example of nano-drug delivery systems and have been successfully applied in clinical practice. They offer promising application prospects and numerous advantages. Compared with traditional dosage forms, their advantages include increasing the solubility of poorly soluble drugs, extending their half-life in vivo, enabling tumor targeting, improving drug pharmacokinetic properties, enhancing drug efficacy, and mitigating adverse reactions. Since the successful launch of Doxil in 1995, research on liposomes has matured, resulting in significant breakthroughs in anti-tumor, anti-fungal, analgesic, and gene therapy.

[0006] Currently, there are no reports on the research of toremifene citrate liposomes. Summary of the Invention

[0007] To overcome the deficiencies of the prior art, the present invention provides a toremifene citrate liposome with high drug loading, high encapsulation efficiency and high stability, and also provides a preparation prepared from the toremifene citrate liposome, which improves the solubility and stability of the toremifene citrate preparation.

[0008] Specifically, the technical solution of the present invention is:

[0009] The first object of the present invention is to provide a toremifene citrate liposome, wherein the toremifene citrate liposome is composed of the following components by weight:

[0010] 10 parts by weight of toremifene citrate

[0011] 10-40 parts by weight of peanut lecithin

[0012] 5-10 parts by weight of cholesterol

[0013] 0.5-2 parts by weight of calcium propionate.

[0014] In a preferred embodiment, the toremifene citrate liposomes are composed of the following components by weight:

[0015] 10 parts by weight of toremifene citrate

[0016] 25 parts by weight of peanut lecithin

[0017] 8 parts by weight of cholesterol

[0018] 1 part by weight of calcium propionate.

[0019] A second object of the present invention is to provide a method for preparing the toremifene citrate liposome, comprising the following steps:

[0020] (1) Peanut lecithin and cholesterol were dissolved in an organic solvent, and the mixture was placed in a water bath at 45°C to 55°C for rotary evaporation to remove 1 / 2 of the organic solvent. Calcium propionate and toremifene citrate were then added, and the mixture was continued to be rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded lecithin film.

[0021] (2) Adding a buffer solution with a pH of 5.0 to 6.5 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure to obtain toremifene citrate liposomes.

[0022] In many embodiments, the organic solvent is selected from one of chloroform, methanol, and ethanol.

[0023] In a preferred embodiment, the organic solvent is chloroform.

[0024] In various embodiments, the buffer solution is selected from one of a phosphate buffer solution, a citrate buffer solution, and a carbonate buffer solution.

[0025] In a preferred embodiment, the buffer solution is a phosphate buffer solution.

[0026] In a preferred embodiment, the pH of the buffer solution is 5.8.

[0027] The third object of the present invention is to provide a toremifene citrate preparation, which is composed of the above-mentioned toremifene citrate liposomes and pharmaceutically acceptable excipients.

[0028] In various embodiments, the preparation is a tablet, a capsule, or a granule.

[0029] The present invention also provides the use of the toremifene citrate liposome in preparing a drug for treating tumors.

[0030] Compared with the prior art, the technical effects of the present invention are:

[0031] 1. The present invention utilizes liposome technology to encapsulate toremifene citrate in a lipid carrier. By optimizing lipid materials and preparation methods, a toremifene citrate liposome with high drug loading, high encapsulation efficiency and high stability is provided.

[0032] 2. The present invention further prepares a toremifene citrate preparation by combining the aforementioned toremifene citrate liposomes with pharmaceutically acceptable excipients. Dissolution and related substance testing demonstrates that the toremifene citrate preparation provided by the present invention exhibits stable, uniform, and rapid dissolution, thereby improving oral bioavailability, reducing impurity content, and enhancing formulation stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1: Electron micrograph of embodiment 2 toremifene citrate liposome

[0034] Figure 2 : Dissolution curves of Examples 5 to 7, Comparative Examples 5 and 6, and commercially available toremifene citrate tablets

[0035] Figure 3 : Changes in the contents of related substances in Examples 5 to 7, Comparative Examples 5 and 6, and commercially available toremifene citrate tablets Implementation Method

[0036] Example 1 Preparation of peanut lecithin

[0037] (1) Add 0.05% cellulase and water to the peanut oil foot and enzymatically hydrolyze for 1.5 h. Then add 0.08% protein hydrolase and continue enzymatic hydrolysis for 1 h. After ultrasonic treatment at 55 °C for 25 min, dry and dehydrate to obtain dehydrated peanut oil foot.

[0038] (2) The dehydrated peanut oil foot was placed in an extraction cell, and CO2 was introduced at a flow rate of 2 mL / min. At a temperature of 35 °C and a pressure of 30 MPa, static extraction was performed for 10 min, and then dynamic extraction was performed for 20 min.

[0039] (3) Pre-mix ethanol with CO2, with the added amount of ethanol being 2%. Then, introduce the CO2 mixed with ethanol at a flow rate of 2 mL / min. At 45°C and a pressure of 35 MPa, extract the product of step (1) for 150 min, separate and recover the carbon dioxide and ethanol, and obtain peanut lecithin.

[0040] Example 2 The formula and preparation method of toremifene citrate liposomes are as follows

[0041] formula:

[0042] 10 parts by weight of toremifene citrate

[0043] 25 parts by weight of peanut lecithin

[0044] 8 parts by weight of cholesterol

[0045] 1 part by weight of calcium propionate.

[0046] Preparation method:

[0047] (1) Peanut lecithin and cholesterol were dissolved in chloroform, and the mixture was placed in a water bath at 45°C to 55°C for rotary evaporation to remove 1 / 2 of the organic solvent. Calcium propionate and toremifene citrate were then added, and the mixture was continued to be rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded lecithin film.

[0048] (2) Adding a phosphate buffer solution of pH=5.8 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure to obtain toremifene citrate liposomes.

[0049] Example 3 The formula and preparation method of toremifene citrate liposomes are as follows

[0050] formula:

[0051] 10 parts by weight of toremifene citrate

[0052] 10 parts by weight of peanut lecithin

[0053] 5 parts by weight of cholesterol

[0054] 0.5 parts by weight of calcium propionate.

[0055] Preparation method:

[0056] (1) Peanut lecithin and cholesterol were dissolved in methanol and rotary evaporated in a water bath at 45°C to 55°C to remove 1 / 2 of the organic solvent. Calcium propionate and toremifene citrate were then added and rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded lecithin film.

[0057] (2) Adding a citrate buffer solution of pH=5.0 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure emulsification to obtain toremifene citrate liposomes.

[0058] Example 4 The formula and preparation method of toremifene citrate liposomes are as follows

[0059] formula:

[0060] 10 parts by weight of toremifene citrate

[0061] 40 parts by weight of peanut lecithin

[0062] 10 parts by weight of cholesterol

[0063] 2 parts by weight of calcium propionate.

[0064] Preparation method:

[0065] (1) Peanut lecithin and cholesterol were dissolved in ethanol and rotary evaporated in a water bath at 45°C to 55°C to remove 1 / 2 of the organic solvent. Calcium propionate and toremifene citrate were then added and rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded lecithin film.

[0066] (2) Adding a carbonate buffer solution with a pH of 6.5 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure to obtain toremifene citrate liposomes.

[0067] Comparative Example 1 The formula and preparation method of toremifene citrate liposomes are as follows

[0068] formula:

[0069] 10 parts by weight of toremifene citrate

[0070] 25 parts by weight of peanut lecithin

[0071] 8 parts by weight of cholesterol.

[0072] Preparation method:

[0073] (1) Peanut lecithin and cholesterol were dissolved in chloroform, and the mixture was rotary evaporated in a water bath at 45°C to 55°C to remove 1 / 2 of the organic solvent. Toremifene citrate was then added and the mixture was rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded phospholipid film.

[0074] (2) Adding a phosphate buffer solution of pH=5.8 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure to obtain toremifene citrate liposomes.

[0075] Comparative Example 2 The formula and preparation method of toremifene citrate liposomes are as follows

[0076] formula:

[0077] 10 parts by weight of toremifene citrate

[0078] 25 parts by weight of peanut lecithin

[0079] 8 parts by weight of cholesterol

[0080] 4 parts by weight of calcium propionate.

[0081] Preparation method:

[0082] (1) Peanut lecithin and cholesterol were dissolved in chloroform, and the mixture was placed in a water bath at 45°C to 55°C for rotary evaporation to remove 1 / 2 of the organic solvent. Calcium propionate and toremifene citrate were then added, and the mixture was continued to be rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded lecithin film.

[0083] (2) Adding a phosphate buffer solution of pH=5.8 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure to obtain toremifene citrate liposomes.

[0084] Comparative Example 3 The formula and preparation method of toremifene citrate liposomes are as follows

[0085] formula:

[0086] 10 parts by weight of toremifene citrate

[0087] 25 parts by weight of peanut lecithin

[0088] 8 parts by weight of cholesterol

[0089] 1 part by weight of potassium sorbate.

[0090] Preparation method:

[0091] (1) Peanut lecithin and cholesterol were dissolved in chloroform, and the mixture was placed in a water bath at 45°C to 55°C for rotary evaporation to remove 1 / 2 of the organic solvent. Potassium sorbate and toremifene citrate were then added, and the mixture was continued to be rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded lecithin film.

[0092] (2) Adding a phosphate buffer solution of pH=5.8 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure to obtain toremifene citrate liposomes.

[0093] Comparative Example 4 The formula and preparation method of toremifene citrate liposomes are as follows

[0094] formula:

[0095] 10 parts by weight of toremifene citrate

[0096] 25 parts by weight of peanut lecithin

[0097] 8 parts by weight of cholesterol

[0098] 1 part by weight of calcium propionate.

[0099] Preparation method:

[0100] (1) Peanut lecithin, cholesterol, and calcium propionate were dissolved in chloroform and rotary evaporated in a water bath at 45°C to 55°C. Toremifene citrate was then added and rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded phospholipid film.

[0101] (2) Adding a phosphate buffer solution of pH=5.8 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure to obtain toremifene citrate liposomes.

[0102] Example 5 The formula and preparation method of Toremifene Citrate Tablets (100 tablets) are as follows

[0103] The toremifene citrate liposomes of Example 1 were filtered and dried, 210 g of which was taken and sieved, and 30 g of microcrystalline cellulose, 120 g of starch, 20 g of sodium starch glycolate, and 5 g of talc were added and tableted to obtain toremifene citrate tablets.

[0104] Example 6 The formula and preparation method of Toremifene Citrate Tablets (100 tablets) are as follows

[0105] The toremifene citrate liposomes of Example 2 were filtered and dried, 199 g of which was taken and sieved, 80 g of starch, 60 g of mannitol, 15 g of low-substituted hydroxypropyl cellulose, and 2 g of magnesium stearate were added, and tablets were obtained to obtain toremifene citrate tablets.

[0106] Example 7 The formula and preparation method of Toremifene Citrate Tablets (100 tablets) are as follows

[0107] The toremifene citrate liposomes of Example 3 were filtered and dried, 279 g of which was taken and sieved, and appropriate amounts of 10 g of sodium carboxymethyl cellulose, 80 g of starch, 10 g of cross-linked polyvinylpyrrolidone, and 1 g of micropowdered silica gel were added and tableted to obtain toremifene citrate tablets.

[0108] Example 8 The formula and preparation method of Toremifene Citrate Capsules (100 capsules) are as follows

[0109] The toremifene citrate liposomes of Example 1 were filtered and dried, 210 g of which was taken and sieved, and 20 g of microcrystalline cellulose, 80 g of starch, 15 g of sodium starch glycolate, and 5 g of talc were added and filled into capsules to obtain toremifene citrate capsules.

[0110] Example 9 The formula and preparation method of Toremifene Citrate Granules (100 bags) are as follows

[0111] The toremifene citrate liposomes of Example 1 were filtered and dried, 210 g of which was taken and sieved, and 50 g of microcrystalline cellulose, 150 g of starch, 5 g of talc, and 15 g of sodium starch glycolate were added and mixed. An appropriate amount of water was added, and the mixture was wet granulated, dried, and granulated to obtain toremifene citrate granules.

[0112] Comparative Example 5 The formula and preparation method of Toremifene Citrate Tablets are as follows

[0113] The toremifene citrate liposomes prepared in Comparative Example 1 were filtered and dried, and appropriate amounts of microcrystalline cellulose, starch, sodium starch glycolate, and talc were added and tableted to obtain toremifene citrate tablets.

[0114] Comparative Example 6 The formula and preparation method of Toremifene Citrate Tablets are as follows

[0115] The toremifene citrate liposomes of Example 3 were filtered and dried, and appropriate amounts of microcrystalline cellulose, starch, sodium starch glycolate, and talc were added and tableted to obtain toremifene citrate tablets.

[0116] Toremifene citrate tablets (H20130705) available on the market Example

[0117] 1. Quality evaluation of toremifene citrate liposomes

[0118] 1. Toremifene citrate liposome drug loading in Examples 2-4 and Comparative Examples 1-5

[0119] The toremifene citrate encapsulated in the toremifene citrate liposomes was measured, and the mass ratio of the toremifene citrate liposomes was calculated to calculate the drug loading.

[0120] Table 1 Drug loading of toremifene citrate liposomes in Examples 2-4 and Comparative Examples 1-5

[0121]

[0122] 2. Toremifene Citrate Liposome Encapsulation Efficiency in Examples 2-4 and Comparative Examples 1-5

[0123] The toremifene citrate liposomes were separated from the free toremifene citrate by dialysis, and the encapsulation efficiency was determined.

[0124] Table 2 Encapsulation efficiency of toremifene citrate liposomes in Examples 2 to 4 and Comparative Examples 1 to 5

[0125]

[0126] 3. Toremifene citrate liposome leakage rate in Examples 2-4 and Comparative Examples 1-5

[0127] The cells were stored at 40±2°C and 75%±5% relative humidity for 1 month and 6 months. The amount of toremifene leaked into the medium after storage and the amount of toremifene encapsulated in the liposomes before storage were measured, and the leakage rate was calculated.

[0128] Table 3 Leakage rate of toremifene citrate liposomes in Examples 2 to 4 and Comparative Examples 1 to 5

[0129]

[0130] In summary, the toremifene citrate liposomes of Examples 2 to 4 and Comparative Examples 1 to 5 were evaluated in terms of drug loading, encapsulation efficiency, and leakage rate. The results showed that the toremifene citrate liposomes of Examples 2 to 4 of the present invention had a high drug loading capacity, which could improve the drug delivery efficiency of the lipid carrier for toremifene citrate; the toremifene citrate liposomes of Examples 2 to 4 of the present invention had a high encapsulation efficiency, which improved the efficiency of toremifene citrate entrapment by the toremifene citrate liposomes; and the toremifene citrate liposomes of Examples 2 to 4 of the present invention had a low leakage rate, indicating that they were highly stable and could extend their shelf life.

[0131] 2. Quality Evaluation of Toremifene Citrate Tablets

[0132] 1. Dissolution of Toremifene Citrate Tablets

[0133] According to the dissolution test method of the 2020 edition of the Chinese Pharmacopoeia, 0.1 mol / L HCl was used as the medium, the rotation speed was 100 r / min, and the temperature was (37±0.5)°C. 1 mL of the dissolution solution was collected at 1, 2, 5, 8, 10, 15, 30, 45, and 60 min, respectively, and filtered through a 0.8 mm filter membrane. The filtrate was diluted 10 times and then injected for determination.

[0134] Figure 2 The dissolution curves of Examples 5 to 7, Comparative Examples 5 and 6, and commercially available toremifene citrate tablets are shown. The results show that the toremifene citrate tablets of the present invention dissolve stably, uniformly, and rapidly in 0.1 mol / L HCl, with a dissolution percentage of 90% at 60 min, thereby improving oral bioavailability.

[0135] 2. Content of related substances in toremifene citrate tablets

[0136] Take appropriate amounts of Examples 5-7, Comparative Examples 5 and 6, and commercially available toremifene citrate tablets, grind them, dissolve them in mobile phase, and dilute them to a solution containing 1.0 mg per 1 mL. This will serve as the test solution. Accurately measure an appropriate amount and dilute it with mobile phase to a solution containing 10 μg per 1 mL. This will serve as the control solution. High-performance liquid chromatography (HPLC) was performed according to General Method 0512, using octadecylsilane bonded silica gel as the filler, methanol-water-triethylamine (930:69:1) as the mobile phase, and a detection wavelength of 240 nm. The theoretical plate number, calculated based on the toremifene peak, should be no less than 1500. Accurately measure 20 μl of each solution and inject them into a liquid chromatograph. Record the chromatogram until twice the retention time of the main component peak. If impurity peaks are present in the chromatogram of the test solution, the sum of the areas of the impurity peaks must not exceed 1.0% of the main peak area of the control solution.

[0137] Among them, the accelerated test conditions are: temperature 40℃±2%, relative humidity: 75%±5%.

[0138] Figure 3 The changes in the content of related substances in Examples 5 to 7, Comparative Examples 5 and 6, and commercially available toremifene citrate tablets show that the toremifene citrate tablets of the present invention have high stability and low content of related substances.

Claims

1. A toremifene citrate liposome, characterized in that: The toremifene citrate liposome is composed of the following ingredients by weight: 10 parts by weight of toremifene citrate 10-40 parts by weight of peanut lecithin 5-10 parts by weight of cholesterol 0.5-2 parts by weight of calcium propionate; The preparation method of the toremifene citrate liposome comprises the following steps: (1) Peanut lecithin and cholesterol were dissolved in an organic solvent, and the mixture was placed in a water bath at 45°C to 55°C for rotary evaporation to remove 1 / 2 of the organic solvent. Calcium propionate and toremifene citrate were then added, and the mixture was continued to be rotary evaporated in a water bath at 50°C to 60°C to form a drug-loaded lecithin film. (2) adding a buffer solution with a pH of 5.0 to 6.5 to the drug-loaded phospholipid film prepared in step (1) for hydration, and homogenizing by ultrasonication and high pressure emulsification to obtain toremifene citrate liposomes; The organic solvent is selected from at least one of chloroform, methanol, and ethanol; The buffer solution is selected from at least one of a phosphate buffer solution, a citrate buffer solution, and a carbonate buffer solution; The preparation method of the peanut lecithin is: (1) Add 0.05% cellulase and water to the peanut oil foot and enzymatically hydrolyze for 1.5 h. Then add 0.08% protein hydrolase and continue enzymatic hydrolysis for 1 h. After ultrasonic treatment at 55 °C for 25 min, dry and dehydrate to obtain dehydrated peanut oil foot. (2) Place the dehydrated peanut oil foot in the extraction cell and introduce CO2 at a flow rate of 2 ml / min. At a temperature of 35 °C and a pressure of 30 MPa, perform static extraction for 10 min and then dynamic extraction for 20 min. (3) Pre-mix ethanol with CO2 in an amount of 2% ethanol, introduce the CO2 mixed with ethanol at a flow rate of 2 ml / min, extract the product of step (1) for 150 min at 45°C and a pressure of 35 MPa, separate and recover the carbon dioxide and ethanol, and obtain peanut lecithin.

2. The toremifene citrate liposome according to claim 1, wherein The toremifene citrate liposome is composed of the following ingredients by weight: 10 parts by weight of toremifene citrate 25 parts by weight of peanut lecithin 8 parts by weight of cholesterol 1 part by weight of calcium propionate.

3. The toremifene citrate liposome according to claim 1, wherein The organic solvent is chloroform.

4. The toremifene citrate liposome according to claim 1, wherein the buffer solution is a phosphate buffer solution.

5. The toremifene citrate liposome according to claim 1, wherein The pH of the buffer solution is 5.8.

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