CN105078892B - The preparation method of albendazole thermal sensitive liposome - Google Patents

Abstract

The invention discloses an albendazole thermosensitive liposome, which consists of dipalmitoylphosphatidylcholine, monopalmitoylphosphatidylcholine, distearoylphosphatidylglycerol and albendazole. The preparation process is as follows: first put the above four raw materials into a round-bottomed flask in proportion, then add the chloroform-methanol mixed solution, remove the chloroform-methanol mixed solution on a rotary evaporator after dissolving; continue to add phosphate buffer solution, and hydrate to obtain milky white liquid; then transfer the milky white liquid to a centrifuge tube, and use an ultrasonic cell disruptor to process it for 150 seconds; finally filter, and freeze-dry the filtrate to obtain a finished product. The remarkable effect of the present invention is that ABZ is encapsulated in the heat-sensitive liposome carrier to increase the solubility of the drug, slow down the release of the drug, enhance the liver targeting, and finally achieve the purpose of improving bioavailability; in addition, the present invention The obtained albendazole thermosensitive liposome (TSL‑ABZ) has a high significant encapsulation efficiency and ABZ drug loading.

Description

The preparation method of albendazole thermosensitive liposome

technical field

The invention relates to a preparation method of a new dosage form of albendazole, in particular to a preparation method of albendazole thermosensitive liposome.

Background technique

Albendazole (ABZ), also known as Changchongqing, chemical name 5-propylthio-1H-benzimidazole-2-carbamate methyl ester, is a broad-spectrum antiparasitic drug of benzimidazoles, But because it is insoluble in water, the intestinal absorption rate and bioavailability are very low. Traditionally, albendazole is in the form of powder or tablet. The cure rate of this common dosage form for non-intestinal parasitic diseases is only about 30%, and the concentration of liver drugs is relatively low. Someone proposed a kind of albendazole liposome afterwards, but adopt lecithin as raw material in its scheme, the albendazole liposome that prepares does not have heat sensitivity.

Later, the researchers conducted a study on high-intensity focused ultrasound-induced albendazole thermosensitive liposomes to kill protoscoleiae, and found that HIFU combined with albendazole thermosensitive lipid (TSL-ABZ) had a The killing effect is better than that of HIFU irradiation alone. When the HIFU power is constant, the death rate of protoscolums increases with the increase of drug concentration within a certain range of TSL-ABZ concentration; this study aims to explore the effect of high intensity focused ultrasound (HIFU) on the Effect of azole thermosensitive liposomes (Albendazolethermosensitive liposomes, TSL-ABZ) on killing protoscoleiae.

The researchers set up sham ultrasound groups, pure HIFU group, HIFU+ABZ water 100μL group, HIFU+TSL-ABZ100μL group, HIFU+TSL-ABZ 200μL group, HIFU+TSL-ABZ 300μL group, HIFU+TSL-ABZ 600μL group group, HIFU+TSL-ABZ 1000μL group, the experimental group was 1mL of protocephala suspension and 1mL of PBS buffer solution containing different concentrations of liposomes, each group was irradiated with HIFU 75W×12s, and the suspension was processed after treatment. Stain with eosin, observe the morphological changes of protoscoleia under a light microscope and count the mortality rate; fix the sections of protoscoleiae in agarose, and observe with HE staining. The results showed that after combined use of albendazole thermosensitive liposomes and high-intensity focused ultrasound, morphological observation showed that the cortex was severely damaged, calcium particles disappeared, vacuolation was obvious, head hooks fell off, and some structures even disappeared, and the mortality rate was high . It shows that the combination of albendazole thermosensitive liposome and high-intensity focused ultrasound has a good killing effect on protoscoleiae.

Contents of the invention

The object of the present invention is to provide a kind of preparation method of albendazole thermosensitive liposome.

The purpose of the invention is achieved in this way:

A kind of preparation method of albendazole thermosensitive liposome, described albendazole thermosensitive liposome is made up of the raw material of following mass parts:

Dipalmitoylphosphatidylcholine 60-70 parts,

Monopalmitoylphosphatidylcholine 5-9 parts,

3-8 parts of distearoylphosphatidylglycerol,

Albendazole 1.5-5 parts;

The gist of it is to follow these steps:

Step 1. Put the dipalmitoylphosphatidylcholine, monopalmitoylphosphatidylcholine, distearoylphosphatidylglycerol and albendazole into the round-bottomed flask successively according to the proportion, and then add the Add the chloroform-methanol mixture, avoid light and fully dissolve;

Step 2, the round-bottomed flask is placed on a rotary evaporator, the chloroform-methanol mixture is removed, and a white film is obtained on the inner wall of the round-bottomed flask;

Step 3, adding a phosphate buffer solution of pH=5 to 6 into the round bottom flask, hydrating for 30 minutes to obtain milky white liquid;

Step 4: Transfer the milky white liquid to a centrifuge tube, use an ultrasonic cell breaker to treat it for 150 seconds, pass it through a 0.22 μm microporous membrane, and finally freeze-dry the obtained filtrate to obtain the finished product.

As a preferred technical scheme, the mass parts of each raw material in the above-mentioned method are:

Dipalmitoylphosphatidylcholine 65-67 parts,

Monopalmitoylphosphatidylcholine 6-8 parts,

4-6 parts of distearoylphosphatidylglycerol,

3 to 4 parts of albendazole.

As a preferred technical solution, in the third step above, the pH of the phosphate buffer solution is 5.3.

As a preferred technical solution, in the above step 4, the ultrasonic power of the ultrasonic cell disruptor is 50%, and the ultrasonic time is 15 minutes.

Beneficial effect: adopting the albendazole thermosensitive liposome and the preparation method thereof of the present invention, ABZ is encapsulated in the thermosensitive liposome carrier, so as to improve the solubility of the drug, slow down the release of the drug, and enhance the liver targeting , and finally achieve the purpose of improving bioavailability; in addition, the albendazole thermosensitive liposome (TSL-ABZ) obtained by the present invention has higher significant encapsulation efficiency and ABZ drug loading capacity.

Detailed ways

The present invention will be further described below in conjunction with embodiment.

A kind of albendazole thermosensitive liposome is made up of the raw material of following mass parts, as shown in table 1:

Table 1, albendazole thermosensitive liposome raw material composition (unit: parts by mass)

A kind of preparation method of albendazole thermosensitive liposome, carries out according to the following steps:

Step 1. Put the dipalmitoylphosphatidylcholine, monopalmitoylphosphatidylcholine, distearoylphosphatidylglycerol and albendazole into the round-bottomed flask successively according to the proportion, and then add the Add the chloroform-methanol mixture, avoid light and fully dissolve;

Step 2, the round-bottomed flask is placed on a rotary evaporator, the chloroform-methanol mixture is removed, and a white film is obtained on the inner wall of the round-bottomed flask;

Step 3, adding a phosphate buffer solution of pH=5 to 6 into the round bottom flask, hydrating for 30 minutes to obtain milky white liquid;

The pH of the phosphate buffer can also be 5.3.

Step 4, transfer the milky white liquid to a centrifuge tube, and use an ultrasonic cell disruptor to process it for 150s. The ultrasonic power of the ultrasonic cell disruptor is 50%, the ultrasonic time is 15min, and then pass through a 0.22 μm microporous filter membrane, and finally The obtained filtrate is freeze-dried to obtain the finished product.

Below in conjunction with test example the present invention will be further described:

Taking Example 1 in Table 1 as an example, the TSL-ABZ obtained in Example 1 is the test group 1, and the following TSL-ABZ control group is set, as shown in Table 2:

Table 2, TSL-ABZ control group raw material composition (unit: mass parts)

The encapsulation efficiency (%) and ABZ drug loading (%) of the test group 1 and the control group 1-1 to 1-6 were tested respectively, and the test results are shown in Table 3:

Encapsulation efficiency (%), ABZ drug loading (%) of table 3, test group and control group

It can be seen from Table 3 that the encapsulation efficiency and ABZ drug loading of the TSL-ABZ of the test group 1 were significantly improved compared with those of the control groups 1-1 to 1-6.

According to the above method, take Example 2, Example 3 and Example 4 as test groups 2, 3, and 4 respectively, and set up the control group 2-1~2-6, the control group 3-1~3-6 and the control group accordingly. Groups 4-1 to 4-6, the analysis of test results showed that the encapsulation efficiency of TSL-ABZ in test groups 2, 3, and 4 were all greater than 79%, and the drug loading of ABZ was greater than 2.1%, and the control group 2-1 The encapsulation efficiencies of TSL-ABZ of ~2-6, control groups 3-1-3-6 and control groups 4-1-4-6 were all less than 46%, and the drug loads of ABZ were all less than 1.26%.

Adopting the present invention can significantly improve the encapsulation efficiency and ABZ drug loading capacity.

Finally, it should be noted that the above description is only a preferred embodiment of the present invention, and those of ordinary skill in the art can make a variety of similar implementations under the inspiration of the present invention without violating the purpose and claims of the present invention. It means that such transformations all fall within the protection scope of the present invention.

Claims (4)

1. a kind of preparation method of albendazole thermal sensitive liposome, the albendazole thermal sensitive liposome is by following mass fraction Raw material forms：

60~70 parts of dipalmitoylphosphatidylcholine,

5~9 parts of monopalmitoylphosphatidylcholine,

3~8 parts of distearoylphosphatidylglycerol,

1.5~5 parts of albendazole；

It is characterized in that carry out according to the following steps：

Step 1: by the dipalmitoylphosphatidylcholine, monopalmitoylphosphatidylcholine, distearoylphosphatidylglycerol and Ah Parbendazole is proportionally put into round-bottomed flask successively, then chloroform-methanol mixed liquor is added into the round-bottomed flask, and lucifuge is filled Divide dissolving；

Step 2: the round-bottomed flask is placed on Rotary Evaporators, chloroform-methanol mixed liquor is removed, in round-bottomed flask Wall obtains white film；

Step 3: adding the phosphate buffer of pH=5~6 into the round-bottomed flask, aquation 30min, obtains milky white liquid；

Step 4: the milky white liquid is transferred in centrifuge tube, using ultrasonic cell disruption instrument handle after 150s after 0.22 μm of miillpore filter, obtained filtrate is finally freezed, get product.

2. the preparation method of albendazole thermal sensitive liposome according to claim 1, it is characterised in that：The matter of the raw material Measuring number is：

65~67 parts of dipalmitoylphosphatidylcholine,

6~8 parts of monopalmitoylphosphatidylcholine,

4~6 parts of distearoylphosphatidylglycerol,

3~4 parts of albendazole.

3. the preparation method of albendazole thermal sensitive liposome according to claim 1 or 2, it is characterised in that：The step In three, the pH=5.3 of the phosphate buffer.

4. the preparation method of albendazole thermal sensitive liposome according to claim 3, it is characterised in that：The step 4 In, the ultrasonic power of the ultrasonic cell disruption instrument is 50%, ultrasonic time 15min.