CN107362143B - Nifedipine proliposome and preparation method thereof - Google Patents

Abstract

The invention discloses a nifedipine proliposome, and also provides a new PEI-STA polymer, which plays a synergistic effect with DSPE-PEG2000 to improve the nifedipine proliposome in the intestinal tract. Compared with the nifedipine crude drug, the present invention obviously improves the solubility of the drug by more than 50 times, and the photostability is improved by 2.53 times, which is convenient for storage and transportation. The pharmacokinetic experiments of nifedipine proliposomes proved that increasing the bioavailability of nifedipine by 10.2 times can improve the solubility, stability and bioavailability of nifedipine, and prolong the nifedipine in vivo. Clearance time, reduce dosing frequency. It is a promising drug for the treatment of hypertension.

Description

A kind of nifedipine proliposome and preparation method thereof

Technical field:

The invention discloses a nifedipine proliposome, and also provides a preparation process of the drug, belonging to the technical field of medical pharmacy.

technical background:

Nifedipine is a pale yellow crystalline powder, odorless and tasteless, and is a calcium channel blocker for the treatment of hypertension. However, nifedipine has poor water solubility, low oral bioavailability, significant liver first-pass effect, and elimination cycle in the body. It is short, the frequency of administration is high, the individual differences are obvious, and the fluctuation of plasma drug concentration will be accompanied by different degrees of side effects, so users often feel dizzy during the first few doses. Common side effects include headache, fatigue, cough, shortness of breath, drowsiness, bradycardia, etc. In severe cases, it can lead to a rapid drop in blood pressure, loss of consciousness, cerebral ischemic infarction, myocardial infarction, complete heart obstruction, and even death. In addition, nifedipine is easily decomposed when exposed to ultraviolet light, and will be decomposed into nitroso derivatives and nitro derivatives when exposed to ultraviolet light, which is not conducive to use and storage.

There are two main obstacles to the absorption of nifedipine in the intestinal tract (1) There is a hydrophilic mucus layer in the intestinal tract, and nifedipine is a hydrophobic drug, which is difficult to penetrate the mucus layer barrier in the intestinal tract and is not easily absorbed by the intestinal tract (2 ) If nifedipine is made into a common liposome preparation, although it can penetrate the mucus layer, its ability to penetrate the intestinal epithelial cell barrier is weak, and the ability to improve the bioavailability of nifedipine is limited.

Invention content:

The invention discloses a nifedipine proliposome, which can improve the solubility, stability and bioavailability of nifedipine, prolong the clearance time of nifedipine in the body, and reduce the frequency of administration.

The invention also discloses the preparation method of the above nifedipine proliposome drug, which can improve the bioavailability of nifedipine and give full play to the drug effect, reduce side effects and prolong the action time. It is a promising drug for the treatment of hypertension.

A nifedipine proliposome disclosed in the invention is characterized in that it is prepared from the following raw materials in proportion by weight and number:

2-10 parts of HSPC, 0.2-3 parts of sodium deoxycholate, 2-8 parts of nifedipine, 0.2-5 parts of polyethyleneimine-linked stearic acid (PEI-STA), 1-7 parts of DSPE-PEG2000 , 4-20 parts of spray drying protective agent, and the rest are preparation accessories.

The preparation method of a kind of nifedipine proliposome of the present invention comprises the following steps:

1) Synthesis of polyethyleneimine-linked stearic acid (PEI-STA):

Dissolve 1-8 parts of polyethyleneimine with a molecular weight of 600-70000 and 0.6-18 parts of stearic acid in 40-200 parts of dichloromethane, and carry out under the catalysis of 1-14 parts of EDCI and 0.5-10 parts of NHS condensation reaction;

Described polyethyleneimine-stearic acid structure is as follows:

Wherein, R=-COCH3(CH2)16 or H;

Each polyethyleneimine polymerization unit connects 1-4 stearic acid molecules-COCH3(CH2)16;

2) Weigh HSPC, sodium deoxycholate, nifedipine, polyethyleneimine-linked stearic acid, DSPE-PEG2000 in proportion, dissolve in 300-3000 parts of methanol and ultrasonicate until completely dissolved to form a lipid phase, and quickly inject 3000 - 15000 parts of phosphate buffer solution to form liposomes, add spray drying protective agent to ultrasonically dissolve, and carry out spray drying to obtain nifedipine proliposomes.

The polyethyleneimine involved in the present invention is a water-soluble polymer material of polyamino acids, and is colorless or light yellow viscous liquid; the present invention adopts hydrophobic stearic acid to modify polyethyleneimine to transform it into amphiphilic Because of the cationic properties of PEI-STA, it can enhance the ability of liposomes to penetrate small intestinal epithelial cells and improve the nifedipine liposomes. bioavailability.

In addition, DSPE-PEG2000 in the liposome formulation enhances the hydrophilicity of the liposome and improves the ability of the liposome to penetrate the mucus layer barrier in the intestinal tract. The invention discloses the synergistic effect of DSPE-PEG2000 and a synthetic new material PEI-STA in a nifedipine proliposome pharmaceutical composition to enhance the oral bioavailability of nifedipine.

The following experiments show that a nifedipine proliposome drug disclosed in the present invention can improve the solubility, stability and bioavailability of nifedipine, reduce side effects, and prolong the clearance time of nifedipine in the body, reducing the Dosing frequency.

Solubility: a nifedipine proliposome drug prepared by using the preferred recipe material of the present invention, the encapsulation rate of the nifedipine proliposome prepared in the example was 94.21%. After the nifedipine prolipid proliposome is reconstituted, the solubility of nifedipine in water at 25°C can reach 10 mg/mL, which is much higher than 200 ug/mL of the nifedipine bulk drug.

Light stability: 1mg/mL nifedipine proliposome and nifedipine bulk drug (dissolved in 10% ethanol, 10% Tween 80, 80% normal saline) were sealed and placed in 4500LX light The photostability of the drug was determined by placing it in a light box with a high intensity for 10 days. The results are shown in Table 1.

Table 1. the photostability of nifedipine proliposome and nifedipine bulk drug of the present invention

In vitro release: artificial intestinal fluid and gastric fluid were used to simulate the release of nifedipine proliposomes and raw materials in the gastrointestinal tract, and 9.6 mg of nifedipine proliposomes and raw materials were placed in a pre-treated dialysis bag (Molecular weight cut-off is 8000-12,000Da), seal both ends of the dialysis bag, put it into 200mL of dissolution medium at a temperature of 37±0.5℃, and at a speed of 350r/min, at a fixed time point, take 5mL of the corresponding dissolution medium, and immediately The same volume of release medium was added, and the time points were selected as 0.083, 0.167, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, 24, and 48h, and nifedipine was measured at different time points. , the cumulative release at each time point was calculated, and the release curve was drawn based on this, as shown in Figure 1. The nifedipine proliposome of the present invention exists relatively stably in the stomach, and has a sustained release effect in the intestinal tract.

Pharmacokinetic study of nifedipine proliposome of the present invention in rats:

The 30 rats were randomly divided into 6 groups, the oral administration dose was 6 mg/kg, and the administration groups were respectively the nifedipine proliposomes of the present invention prepared in Example 2 and the nifedipine without DSPE-PEG2000. Proliposomes, nifedipine proliposomes without PEI-STA, nifedipine proliposomes without DSPE-PEG2000 and PEI-STA, nifedipine bulk drug, containing DSPE-PEG2000 and nifedipine proliposomes of polyethyleneimine. At 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, and 36 h after administration, blood was collected from the eye canthus of the two groups of rats, and placed in a centrifuge tube treated with heparin sodium. , add 10 μL of 0.02 mg/mL internal standard nimodipine solution, add 10 μL of 1 mol/L sodium hydroxide, vortex for 1 min, then add ether-trichloromethane (5:1) 600 μL, vortex for 10 min, 10,000 rpm , centrifuge for 10 min, take the upper organic phase and place it in another test tube, dry it under nitrogen flow at 40°C, redissolve the residue in 200 μL of acetonitrile-water (1:1), and determine the content of nifedipine by high performance liquid chromatography, see figure 2.

Conclusion: The pharmacokinetic results are shown in Figure 2. The area under the curve (AUC) of nifedipine proliposomes without DSPE-PEG2000 and PEI-STA in rats is given as nifedipine. 1.51 times that of the bulk drug group; the AUC of the nifedipine proliposome prepared by the present invention is 10.20 times that of the nifedipine bulk drug group; and the nifedipine prolipid without DSPE-PEG2000 The AUC of the nifedipine proliposome was 2.28 times that of the nifedipine bulk drug group; the AUC of the nifedipine proliposome without PEI-STA was 2.21 times that of the nifedipine bulk drug group. The elimination half-life of the nifedipine bulk drug is 1.39h, and the elimination half-life of the nifedipine proliposome of the present invention is 6.30h.

The AUC of nifedipine proliposome containing DSPE-PEG2000 and polyethyleneimine was only 2.86 times that of the nifedipine bulk drug group.

To sum up, the DSPE-PEG2000 and PEI-STA involved in the present invention have a synergistic effect in the preparation of nifedipine proliposomes, and greatly promote the intestinal absorption of nifedipine proliposomes. Compared with the synthesized new material PEI-STA, the unmodified polyethyleneimine does not have the effect of synergistically with DSPE-PEG2000 to improve the bioavailability of nifedipine.

The nifedipine proliposome of the invention can improve the bioavailability of nifedipine, prolong the elimination half-life and stabilize the blood drug concentration.

The nifedipine proliposome drug disclosed in the invention increases the contact area between the drug and the intestinal epithelial cells by increasing the solubility of nifedipine, so as to improve the absorption efficiency, and in addition, the nifedipine proliposome is released in the stomach Slow formation of a colloidal structure, delaying gastric emptying, and improving residence time are beneficial for improving bioavailability, prolonging half-life and stabilizing plasma drug concentrations.

The beneficial effects of the present invention are:

A new PEI-STA polymer is provided, which plays a synergistic effect with DSPE-PEG2000 to improve the absorption effect of nifedipine proliposome in the intestinal tract; The solubility of the drug is more than 50 times, and the photostability is increased by 2.53 times, which is convenient for storage and transportation. The pharmacokinetic experiments of nifedipine proliposomes proved that the bioavailability of nifedipine was increased by 10.2 times.

Description of drawings

Figure 1. Cumulative release rate of nifedipine proliposomes and raw materials in artificial intestinal juice and gastric juice;

Figure 2. The drug-time curve of nifedipine proliposome in rats.

Detailed ways:

The present invention is further described by the following examples, and does not limit the present invention in any way. On the premise of not departing from the technical solutions of the present invention, any changes or changes that are easily realized by those of ordinary skill in the art made by the present invention will be fall within the scope of the claims of the present invention.

Example 1:

Molecular weight is 1 gram of polyethyleneimine of 600, 0.6 gram of stearic acid, reaction catalyst is 1 gram of EDCI, NHS is 0.5 gram, and reaction solvent is 40 milliliters of methylene dichlorides, and the stearic acid ( PEI-STA),

2 g of HSPC, 0.2 g of sodium deoxycholate, 2 g of nifedipine, 0.2 g of polyethyleneimine-linked stearic acid, 1 g of DSPE-PEG2000 dissolved in 300 ml of methanol and sonicated until completely dissolved to form a lipid phase, Quickly inject 3000 ml of phosphate buffer to form liposomes, add 4 grams of spray drying protective agent mannitol to dissolve by ultrasonic, and carry out spray drying to obtain nifedipine proliposomes.

Example 2:

Molecular weight is 8 grams of polyethyleneimine of 70000, 18 grams of stearic acid, reaction catalyst is that EDCI is 14 grams, NHS is 10 grams, and reaction solvent is 200 milliliters of methylene dichlorides, and the stearic acid ( PEI-STA),

10 g of HSPC, 3 g of sodium deoxycholate, 8 g of nifedipine, 5 g of polyethyleneimine-linked stearic acid, and 7 g of DSPE-PEG2000 were dissolved in 3000 ml of methanol and sonicated until completely dissolved to form a lipid phase, Quickly inject 15,000 ml of phosphate buffer to form liposomes, add 20 grams of spray drying protective agent mannitol for ultrasonic dissolution, and carry out spray drying to obtain nifedipine proliposomes.

Example 3:

Molecular weight is 4 grams of polyethyleneimine of 8000, 12 grams of stearic acid, reaction catalyst is that EDCI is 10 grams, NHS is 8 grams, and reaction solvent is 100 milliliters of methylene dichlorides, and the stearic acid ( PEI-STA),

5 g of HSPC, 1 g of sodium deoxycholate, 4 g of nifedipine, 2 g of polyethyleneimine-linked stearic acid, 3 g of DSPE-PEG2000 dissolved in 1000 ml of methanol and sonicated until completely dissolved to form a lipid phase, Quickly inject 10,000 ml of phosphate buffer to form liposomes, add 10 grams of spray drying protective agent mannitol for ultrasonic dissolution, and carry out spray drying to obtain nifedipine proliposomes.

Clinical efficacy test:

The clinical trial used a double-blind method to randomly divide 240 eligible patients with essential hypertension after clinical physical examination into two groups (control group and experimental group). The patients in the experimental group took the nifedipine proliposomes (containing 5 mg of nifedipine) of Examples 1 to 3, once a day. After 1 week of continuous medication, the systolic and diastolic blood pressures of the two groups of patients were measured, and the adverse reactions of the patients during the treatment were counted. The results are shown in Tables 2 and 3.

Table 2. Therapeutic effects of nifedipine proliposome and nifedipine

Before treatment, there was no significant difference in diastolic and systolic blood pressure between the experimental group and the control group (P>0.05). After one week of treatment, the blood pressure of the patients in the experimental group decreased significantly compared with the control group. *Represents the experimental group after treatment compared with the control group after treatment, P<0.01.

Table 3. Adverse reactions of nifedipine proliposome and nifedipine

Compared with the API, nifedipine proliposome has higher bioavailability, so the antihypertensive effect can be improved on the basis of reducing the dosage, and the elimination half-life of nifedipine proliposome is long, and the blood The drug concentration is stable, which can reduce the side effects caused by multiple high-dose administration.

Claims (1)

1. a nifedipine proliposome is characterized in that being made by following proportioning raw material by weight and number ratio: 2-10 parts of HSPC, 0.2-3 parts of sodium deoxycholate, 2-8 parts of nifedipine, 0.2-5 parts of polyethyleneimine-linked stearic acid, 1-7 parts of DSPE-PEG2000, spray drying protective agent 4-20 servings, the rest are preparation excipients; The preparation method of above-mentioned nifedipine proliposome, comprises the following steps: 1) Synthesis of polyethyleneimine-linked stearic acid: Dissolve 1-8 parts of polyethyleneimine with a molecular weight of 600-70000 and 0.6-18 parts of stearic acid in 40-200 parts of dichloromethane, and carry out under the catalysis of 1-14 parts of EDCI and 0.5-10 parts of NHS condensation reaction; 2) Weigh HSPC, sodium deoxycholate, nifedipine, polyethyleneimine-linked stearic acid, DSPE-PEG2000 in proportion, dissolve in 300-3000 parts of methanol and ultrasonicate until completely dissolved to form a lipid phase, and quickly inject 3000 - 15000 parts of phosphate buffer solution to form liposomes, add spray drying protective agent to ultrasonically dissolve, and carry out spray drying to obtain nifedipine proliposomes.

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