CN106924176B - Tamoxifen flexible nano liposome gel and preparation method thereof - Google Patents

Abstract

The invention discloses a tamoxifen flexible nano liposome gel and a preparation method thereof, wherein the gel is prepared from tamoxifen, cholesterol, a phospholipid material, a water-soluble gel matrix, a transdermal penetration enhancer, a surfactant, a preservative, a humectant, a pbs buffer solution, ammonia water, distilled water and ethanol which are used as raw materials according to a certain proportion. The preparation method comprises the following steps: 1) preparing tamoxifen flexible nanoliposome suspension; 2) preparing a blank gel matrix; 3) and uniformly mixing the blank gel matrix and the tamoxifen flexible nano liposome suspension to obtain the tamoxifen flexible nano liposome gel. The gel has high drug loading and encapsulation effects on tamoxifen, and has good stability and transdermal permeability. The preparation method is simple, convenient to operate and low in preparation cost.

Description

A kind of tamoxifen flexible nanoliposome gel and preparation method thereof

technical field

The invention relates to the technical field of pharmaceutical preparations, in particular to a tamoxifen flexible nanoliposome gel and a preparation method thereof.

Background technique

Tamoxifen (TAM) is a selective estrogen receptor modulator (SERM) that interferes with certain activities of estrogen, mimics the effects of other estrogens, and upregulates the production of transforming growth factor beta, which The reduction is associated with the development of malignant tumors and also has a specific inhibitory effect on protein kinase C. These effects all have inhibitory effects on tumor cells that are dependent on estrogen for continued growth. Clinically, it is mostly used for the treatment of breast cancer with progressive development of premenopausal estrogen (ER+) and progesterone receptor (PR+).

Nearly 100 countries in the world use TAM to treat advanced recurrent breast cancer and ovarian cancer, mainly for the treatment and prevention of breast cancer. Patients are prevented from dying because of TAM treatment. Although new drugs for breast cancer endocrine therapy emerge in an endless stream, they cannot shake their status. However, oral TAM has many inevitable side effects, such as fever, endometrial thickening, etc. In clinical practice, drugs Treatment compliance is often not high. If the adverse reactions can be improved on the premise of ensuring the local breast dose, it will undoubtedly be a huge breakthrough. In a sense, it will open a new window for chemotherapy.

The preparations of TAM in clinical application are mostly tablets. TAM is widely distributed in the body, and it kills tumor cells and also damages normal cells. During treatment, it may cause DNA damage, hypertriglyceridemia, bone marrow suppression, ocular toxicity, psychotic episodes, endocrine disorders, and vascular embolism. Sexual diseases, ovarian lesions, endometriosis, etc., and even induce endometrial cancer, increase systemic adverse reactions and affect clinical application. In order to make TAM more concentrated in the tumor site, the preparation research of TAM has become a hot spot. Modern studies have shown that the TAM gel is prepared into a gel, and it is found that the TAM gel and the oral regimen have a similar decrease in the Ki67 labeling index, while the systemic effect is more pronounced. Small. Zhu Ling et al. found that the tamoxifen citrate gel formulation formed a drug reservoir in the mammary gland and its surrounding tissues after percutaneous administration, which had a certain sustained-release property and significantly prolonged the half-life of tamoxifen in plasma. . The TAM magnetic microspheres prepared by Liu Rangru et al. increased its drug concentration at the tumor site and increased its efficacy. The TAM microemulsion external preparation prepared by Zhang Yi et al. It has become the first choice for research to improve the side effects of tamoxifen and improve the efficacy. It is reported that TAM external preparations include ointment and cream, but there is no report of TAM transfer body gel, so it is necessary to improve the existing dosage form. , to prepare a dosage form with constant drug release rate, high bioavailability and less toxic and side effects, which requires the efforts of scientific researchers to explore.

SUMMARY OF THE INVENTION

Based on the above-mentioned prior art, the present invention provides a tamoxifen flexible nanoliposome gel and a preparation method thereof. The gel has high drug loading and encapsulation effect on tamoxifen, And the stability and transdermal permeability are good.

The preparation method is simple, the operation is convenient, and the preparation cost is low.

The technical scheme adopted to realize the above-mentioned purpose of the present invention is:

A tamoxifen flexible nanoliposome gel is prepared from the following raw materials in mass fractions:

A tamoxifen flexible nanoliposome gel is prepared from the following raw materials in mass fractions:

Further, the phospholipid material is one of soybean lecithin, hydrogenated soybean lecithin and dipalmitoyl phosphatidylcholine, or a composite phospholipid of hydrogenated soybean lecithin and soybean lecithin, or hydrogenated soybean lecithin and dipalmitoyl phosphatidylcholine A complex phospholipid of palmitoylphosphatidylcholine.

Further, the water-soluble gel matrix is a cellulose derivative, and the cellulose derivative is methyl cellulose or sodium carboxymethyl cellulose.

Further, the skin penetration enhancer is amine, menthol, lauro azone, poloxamer, sodium lauryl sulfate, fatty acid or fatty acid ester.

Further, the amine is urea; the fatty acid is oleic acid or lauric acid; the fatty acid ester is lauryl lactate, isopropyl myristate, propylene glycol dipelargonate or two sebacate ethyl ester.

Further, described surfactant A is sodium cholate, sodium deoxycholate, fatty acid glyceride, sucrose fatty acid ester, polysorbate or polyoxyethylene fatty acid ester, and described surfactant B is cholic acid Sodium, sodium deoxycholate or fatty acid glycerides.

Further, the preservatives are benzoic acid, sodium benzoate, sorbic acid or parabens, and the parabens are methylparaben, ethylparaben, paraben Propyl or butyl paraben.

Further, the moisturizing agent is glycerin, propylene glycol or sorbitol.

A preparation method of tamoxifen flexible nano-liposome gel, comprising the following steps:

1) Dissolve tamoxifen, cholesterol, phospholipid material, and surfactant A in an organic solvent, wherein the organic solvent is chloroform, ethanol or diethyl ether, obtain solution A, concentrate solution A to remove the organic solvent, add PBS buffer The mass of the PBS buffer added is 1-5% of the mass of the water-soluble gel matrix, and it is hydrated under normal temperature and pressure to obtain a tamoxifen flexible nanoliposome suspension;

2) Mix the water-soluble gel matrix, surfactant B and distilled water homogeneously, and the quality that the distilled water adds is 0.1-1% of the quality of the water-soluble gel matrix to obtain solution B, and then mix the transdermal penetration enhancer, preservative and The humectant is dissolved in ethanol, and the mass of the ethanol added is 7-12% of the mass of the water-soluble gel matrix to obtain solution C. Add solution C to solution B under stirring. After swelling is good, add ammonia water to adjust to neutrality. The added mass is 1-5% of the mass of the water-soluble gel matrix to obtain a blank gel matrix;

3) Mixing the blank gel matrix and the tamoxifen flexible nanoliposome suspension to obtain a tamoxifen flexible nanoliposome gel.

Compared with the prior art, the beneficial effects and advantages of the present invention are:

1. In terms of drug loading and encapsulation efficiency, the present study found that the sample loading of the tamoxifen flexible nanoliposome gel was 0.5-1.2 mg/mL, and the encapsulation efficiency was 85.6-96.0%. The flexible nanoliposome gel of the present invention has higher drug loading and encapsulation effects on tamoxifen.

2. In terms of stability, the present study found that the particle size of the tamoxifen flexible nanoliposome gel is between 50.2±0.12 and 100.8±0.26nm, and the potential is -21.8±0.15～-37.7±0.45 mv, the polydispersity coefficient is 0.1 ± 0.05 ~ 0.18 ± 0.01, put the tamoxifen flexible nanoliposome gel at room temperature and 4 ° C, respectively, and observe the appearance, particle size and particle size within 1, 5, 10, and 15 days. The particle size, potential and drug content of tamoxifen flexible nanoliposome gel were found to increase slightly at room temperature, but the potential did not change significantly. changes, thus indicating that the tamoxifen flexible nanoliposome gel of the present invention has good stability at room temperature and 4°C.

3. In the aspect of in vitro transdermal permeability, the present study found that the percutaneous steady-state permeation rate and intradermal retention of the tamoxifen flexible nanoliposome gel are approximately 3 times that of the ordinary tamoxifen gel. -7 times, indicating that the flexible nanoliposome is a novel carrier that can effectively promote transdermal penetration.

In a word, the present invention uses tamoxifen as a drug and a flexible nanoliposome gel as a carrier, which solves the defects of tamoxifen with large side effects and drug compliance. The tamoxifen flexible nanolipid of the present invention The body gel can continuously release and penetrate the skin within 24 hours, and the drug enters the administration site through the skin, which can maintain a stable local blood drug concentration, reduce the number of patients taking the drug, and increase the patient's compliance. In addition, the tamoxifen flexible nanoliposome gel of the present invention avoids the first-pass effect of oral administration of the drug through the gastrointestinal tract and the liver, and improves the bioavailability. Therefore, the tamoxifen flexible nanoliposome gel preparation of the present invention has a fast absorption rate, a high transdermal amount, has the characteristics of stability and high efficiency, has obvious advantages in clinical application, and can effectively realize the drug for a long time. continuous transdermal penetration, so as to achieve the targeted effect of local administration.

Instruction drawings

Fig. 1 is the percutaneous penetration curve of the tamoxifen flexible nanoliposome gel prepared in Example 1 and the common tamoxifen gel.

Figure 2 is the percutaneous penetration curve of the tamoxifen flexible nanoliposome gel prepared in Example 2 and the common tamoxifen gel.

FIG. 3 is the transdermal penetration curve of the tamoxifen flexible nanoliposome gel prepared in Example 3 and the common tamoxifen gel.

FIG. 4 is a comparison diagram of the skin drug retention of the tamoxifen flexible nanoliposome gel prepared in Example 1 and the common tamoxifen gel.

Figure 5 is a comparison diagram of the skin drug retention of the tamoxifen flexible nanoliposome gel prepared in Example 2 and the common tamoxifen gel.

6 is a comparison diagram of the skin drug retention of the tamoxifen flexible nanoliposome gel prepared in Example 3 and the common tamoxifen gel.

Detailed ways

The present invention will be described in detail below with reference to specific embodiments.

Example 1

A tamoxifen flexible nanoliposome gel is prepared from the following raw materials in mass fractions:

The phospholipid material is a composite phospholipid of hydrogenated soybean lecithin and soybean lecithin, which is denoted as SPC/HSPC (the molar ratio of SPC and HSPC is 3:1).

The preparation method of above-mentioned tamoxifen flexible nano-liposome gel, its steps are:

1) Dissolve tamoxifen, cholesterol, SPC/HSPC and fatty acid glycerides in chloroform to obtain solution A, remove chloroform by rotary evaporation under reduced pressure at 37° C., add PBS buffer (pH=7.4), Hydrate for 15 minutes at room temperature and pressure to obtain a tamoxifen flexible nanoliposome suspension;

2) Mix sodium carboxymethyl cellulose, sodium deoxycholate and distilled water evenly to obtain solution B, then dissolving laurozone, benzoic acid and propylene glycol in ethanol to obtain solution C, and gradually add solution C to the solution In B, stirring at a speed of 500rpm for 15min, after mixing and swelling, ammonia water (25wt%) was added to adjust the pH value to about 7.0 to obtain a blank gel matrix;

3) Mix the blank gel matrix and the tamoxifen flexible nanoliposome suspension, and stir at 500 rpm for 15 minutes to obtain a tamoxifen flexible nanoliposome gel.

Test 1. Transdermal experiment of the tamoxifen flexible nanoliposome gel of the present invention

experiment method:

1) Fix the treated suckling pig skin on the Frank diffusion cell, add PBS buffer (PH=7.4) in the receiving cell, and add tamoxifen flexible nanoliposome gel (prepared in Example 1) in the supply chamber. ), a constant temperature water bath at 37°C and stirring at 300r·min ^-1 at the same time, the receiving solution was taken at 0.5h, 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 48h, and an equal amount of PBS was added. The mobile phase (acetonitrile: water = 60:40) was diluted 10 times, analyzed by high performance liquid chromatography, the content of tamoxifen was determined, and the cumulative transdermal penetration was calculated; The blank gel prepared by the method in step 2 of this example was prepared by mixing and stirring with tamoxifen) as a control, and experiments were carried out under the same conditions, the content of tamoxifen was measured, and the cumulative transdermal penetration was calculated.

2) Remove the suckling pig skin after the skin penetration test in 1), remove the surface drug of the suckling pig skin, rinse the stratum corneum and dermis layer with PBS buffer, wipe the surface moisture, accurately weigh, and then cut the suckling pig into pieces. The skin was placed in a 10 mL volumetric flask, and methanol was ultrasonically extracted for 1 h, and the volume was constant to obtain a skin extract. : water = 60: 40) diluted 10 times, analyzed by high performance liquid chromatography, determined the content of tamoxifen, and calculated the retention amount of tamoxifen per unit mass of skin.

test results:

1. The percutaneous penetration curves of the tamoxifen flexible nanoliposome gel prepared in Example 1 and the common tamoxifen gel are shown in Figure 1. The tamoxifen flexible nanoliposome gel The fitting equation of the transdermal penetration curve of the gel is Q=8.196t-15.935 (r=0.994), and the fitting equation of the common tamoxifen gel is Q=2.1717t-4.03 (r=0.991). The drug was released uniformly for 48 hours, the cumulative permeation amount of the tamoxifen flexible nanoliposome gel prepared in Example 1 was 378.26 μg·cm ^-1 , and the percutaneous steady-state permeation rate J was 8.196ug·cm ^-2 ·h ^{- 1} .

2. The skin drug retention of the tamoxifen flexible nanoliposome gel and ordinary tamoxifen gel prepared in Example 1 is shown in Figure 4. It can be seen from Figure 4 that the preparation in Example 1 The skin drug retention of the tamoxifen flexible nanoliposome gel was 15.96 μg·g ^-1 , and the skin drug retention of the ordinary tamoxifen gel was 5.63 μg·g ^-1 .

It can be seen that the tamoxifen flexible nanoliposome gel of the present invention has a good controlled release drug, and can significantly increase the drug skin retention, thereby prolonging the drug action time, reducing the drug toxicity and side effects, and achieving local treatment.

Example 2

A tamoxifen flexible nanoliposome gel is prepared from the following raw materials in mass fractions:

The phospholipid material is a complex phospholipid of dipalmitoyl phosphatidyl choline and soybean lecithin, which is denoted as DPPC/HSPC (the molar ratio of DPPC and HSPC is 1:3).

The preparation method of above-mentioned tamoxifen flexible nano-liposome gel, its steps are:

1) Dissolve tamoxifen, cholesterol, DPPC/HSPC, and sodium cholate in chloroform to obtain solution A, remove chloroform by rotary evaporation under reduced pressure at 37° C., add PBS buffer (pH=7.4), Hydrate for 15 minutes at room temperature and pressure to obtain a tamoxifen flexible nanoliposome suspension;

2) Mix sodium carboxymethyl cellulose, sodium deoxycholate and distilled water evenly to obtain solution B, then dissolving laurodiazone, benzoic acid and propylene glycol in ethanol to obtain solution C, and gradually add solution C to the solution In B, stirring at 500rpm for 15min, after mixing and swelling, adding ammonia water to adjust the pH value to about 7.0 to obtain a blank gel matrix;

3) Mix the blank gel matrix and the tamoxifen flexible nanoliposome suspension, and stir at 500 rpm for 15 minutes to obtain a tamoxifen flexible nanoliposome gel.

Test 2. Transdermal experiment of the tamoxifen flexible nanoliposome gel of the present invention

experiment method:

1) Fix the treated suckling pig skin on the Frank diffusion cell, add PBS buffer (PH=7.4) in the receiving cell, and add tamoxifen flexible nanoliposome gel (prepared in Example 2) in the supply chamber. ), a constant temperature water bath at 37°C and stirring at 300r·min ^-1 at the same time, the receiving solution was taken at 0.5h, 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 48h, and an equal amount of PBS was added. The mobile phase (acetonitrile: water = 60:40) was diluted 10 times, analyzed by high performance liquid chromatography, the content of tamoxifen was determined, and the cumulative transdermal penetration was calculated; The blank gel prepared by the method in step 2 of this example was prepared by mixing and stirring with tamoxifen) as a control, and experiments were carried out under the same conditions, the content of tamoxifen was measured, and the cumulative transdermal penetration was calculated.

2) Remove the suckling pig skin after the skin penetration test in 1), remove the surface drug of the suckling pig skin, rinse the stratum corneum and dermis layer with PBS buffer, wipe the surface moisture, accurately weigh, and then cut the suckling pig into pieces. The skin was placed in a 10 mL volumetric flask, and methanol was ultrasonically extracted for 1 h, and the volume was constant to obtain a skin extract. : water = 60: 40) diluted 10 times, analyzed by high performance liquid chromatography, determined the content of tamoxifen, and calculated the retention amount of tamoxifen per unit mass of skin.

test results:

1. The percutaneous penetration curves of the tamoxifen flexible nanoliposome gel prepared in Example 2 and the common tamoxifen gel are shown in Figure 2, and the drug was released uniformly for 48 hours. The cumulative permeation amount of the flexible nanoliposome gel of moxifen was 523.87μg·cm ^-1 , and the percutaneous steady-state permeation rate J was 11.383ug·cm ^-2 ·h ^-1 .

2. The skin drug retention of the tamoxifen flexible nanoliposome gel and ordinary tamoxifen gel prepared in Example 2 is shown in Figure 5. It can be seen from Figure 5 that the preparation in Example 2 The skin drug retention of the tamoxifen flexible nanoliposome gel was 11.47μg·g ^-1 , and the skin drug retention of the ordinary tamoxifen gel was 5.43μg·g ^-1 .

It can be seen that the tamoxifen flexible nanoliposome gel of the present invention has a good controlled release drug, and can significantly increase the drug skin retention, thereby prolonging the drug action time, reducing the drug toxicity and side effects, and achieving local treatment.

Example 3

A tamoxifen flexible nanoliposome gel is prepared from the following raw materials in mass fractions:

The preparation method of above-mentioned tamoxifen flexible nano-liposome gel, its steps are:

1) Dissolve tamoxifen, cholesterol, HSPC and polysorbate in chloroform to obtain solution A, remove chloroform by rotary evaporation under reduced pressure at 37°C, add PBS buffer (pH=7.4), and at room temperature Hydrate for 15 min under normal pressure to obtain a tamoxifen flexible nanoliposome suspension;

2) Mix sodium carboxymethyl cellulose, fatty acid glycerides and distilled water to obtain solution B, then dissolve laurofenone, benzoic acid and propylene glycol in ethanol to obtain solution C, and gradually add solution C to solution B , stirring at 500rpm for 15min, after mixing and swelling, adding ammonia water to adjust the pH value to about 7.0 to obtain a blank gel matrix;

3) Mix the blank gel matrix and the tamoxifen flexible nanoliposome suspension, and stir at 500 rpm for 15 minutes to obtain a tamoxifen flexible nanoliposome gel.

Test 3. Transdermal experiment of the tamoxifen flexible nanoliposome gel of the present invention

experiment method:

1) Fix the treated suckling pig skin on the Frank diffusion cell, add PBS buffer (PH=7.4) in the receiving cell, and add the tamoxifen flexible nanoliposome gel (prepared in Example 3) in the supply chamber. ), a constant temperature water bath at 37°C and stirring at 300r·min ^-1 at the same time, the receiving solution was taken at 0.5h, 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 48h, and an equal amount of PBS was added. The mobile phase (acetonitrile: water = 60:40) was diluted 10 times, analyzed by high performance liquid chromatography, the content of tamoxifen was determined, and the cumulative transdermal penetration was calculated; The blank gel prepared by the method in step 2 of this example was prepared by mixing and stirring with tamoxifen) as a control, and experiments were carried out under the same conditions, the content of tamoxifen was measured, and the cumulative transdermal penetration was calculated.

2) Remove the suckling pig skin after the skin penetration test in 1), remove the surface drug of the suckling pig skin, rinse the stratum corneum and dermis layer with PBS buffer, wipe the surface moisture, accurately weigh, and then cut the suckling pig into pieces. The skin was placed in a 10 mL volumetric flask, and methanol was ultrasonically extracted for 1 h, and the volume was constant to obtain a skin extract. : water = 60: 40) diluted 10 times, analyzed by high performance liquid chromatography, determined the content of tamoxifen, and calculated the retention amount of tamoxifen per unit mass of skin.

test results:

1. The percutaneous penetration curves of the tamoxifen flexible nanoliposome gel prepared in Example 3 and the common tamoxifen gel are shown in Figure 3, and the drug was released uniformly for 48 hours. The cumulative permeation amount of the flexible nanoliposome gel of moxifen was 1289.32μg·cm ^-1 , and the steady-state permeation rate J was 27.367ug·cm ^-2 ·h ^-1 .

2. The skin drug retention of the tamoxifen flexible nanoliposome gel and ordinary tamoxifen gel prepared in Example 3 is shown in Figure 6. As can be seen from Figure 6, Example 3 prepared The skin drug retention of tamoxifen flexible nanoliposome gel was 8.27μg·g ^-1 , and the skin drug retention of ordinary tamoxifen gel was 5.13μg·g ^-1 .

It can be seen that the tamoxifen flexible nanoliposome gel of the present invention has a good controlled release drug, and can significantly increase the drug skin retention, thereby prolonging the drug action time, reducing the drug toxicity and side effects, and achieving local treatment.

Claims (4)

1. a tamoxifen flexible nano-liposome gel is characterized in that comprising the raw material of following massfraction:

The water-soluble gel matrix is a cellulose derivative, and the cellulose derivative is methyl cellulose or sodium carboxymethyl cellulose. Described transdermal penetration enhancer is lauroketone; Described surfactant A is sodium cholate, sodium deoxycholate, fatty acid glyceride, sucrose fatty acid ester, polysorbate or polyoxyethylene fatty acid ester, and described surfactant B is sodium cholate, Sodium deoxycholate or fatty acid glycerides; The moisturizing agent is glycerin, propylene glycol or sorbitol. 2. The tamoxifen flexible nanoliposome gel according to claim 1, characterized in that: the phospholipid material is soybean lecithin, hydrogenated soybean lecithin and dipalmitoyl phosphatidylcholine One is either a complex phospholipid of hydrogenated soybean lecithin and soybean lecithin, or a complex phospholipid of hydrogenated soybean lecithin and dipalmitoyl phosphatidylcholine. 3. tamoxifen flexible nanoliposome gel according to claim 1, is characterized in that: described antiseptic is benzoic acid, sodium benzoate, sorbic acid or paraben, described Parabens are methylparaben, ethylparaben, propylparaben or butylparaben. 4. a preparation method of the arbitrary described tamoxifen flexible nano-liposome gel of claim 1-3, is characterized in that comprising the steps: 1) Dissolve tamoxifen, cholesterol, phospholipid material, and surfactant A in an organic solvent, wherein the organic solvent is chloroform, ethanol or diethyl ether, obtain solution A, concentrate solution A to remove the organic solvent, add pbs buffer The added mass of the PBS buffer is 1-5% of the mass of the water-soluble gel matrix, and hydrated at normal temperature and pressure to obtain a tamoxifen flexible nanoliposome suspension; 2) Mix the water-soluble gel matrix, surfactant B and distilled water homogeneously, and the quality that the distilled water adds is 0.1-1% of the quality of the water-soluble gel matrix to obtain solution B, and then mix the transdermal penetration enhancer, preservative and The humectant is dissolved in ethanol, and the mass of the ethanol added is 7-12% of the mass of the water-soluble gel matrix to obtain solution C. Add solution C to solution B under stirring. After swelling is good, add ammonia water to adjust to neutrality. The added mass is 1-5% of the mass of the water-soluble gel matrix to obtain a blank gel matrix; 3) Mixing the blank gel matrix and the tamoxifen flexible nanoliposome suspension to obtain a tamoxifen flexible nanoliposome gel.

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