CN112494429A

CN112494429A - Rupatadine fumarate nanoemulsion and preparation method and application thereof

Info

Publication number: CN112494429A
Application number: CN202011595137.8A
Authority: CN
Inventors: 马燕辉; 潘海群; 侯培宁; 侯宇翔; 戴信敏
Original assignee: Beijing Fahrenheit Kaiyuan Pharmaceutical Technology Co ltd
Current assignee: Beijing Fahrenheit Kaiyuan Pharmaceutical Technology Co ltd
Priority date: 2020-12-29
Filing date: 2020-12-29
Publication date: 2021-03-16
Anticipated expiration: 2040-12-29
Also published as: CN112494429B

Abstract

The invention belongs to the field of medicines, and particularly relates to rupatadine fumarate nanoemulsion as well as a preparation method and application thereof. The rupatadine fumarate nanoemulsion per 100ml comprises the following components: 100 mg-150 mg of rupatadine fumarate; 40-60 g of oil phase; 10 g-30 g of surfactant; 5-15 g of cosurfactant; a proper amount of pH regulator; the volume of water is 100 mL. The nano emulsion provided by the invention does not contain cyclodextrin compounds, has higher safety, uniform particle size distribution, clear and transparent appearance and good fluidity and stability, and can increase the bioavailability by increasing the solubility of rupatadine fumarate.

Description

Rupatadine fumarate nanoemulsion and preparation method and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to rupatadine fumarate nanoemulsion as well as a preparation method and application thereof.

Background

Rupatadine fumarate, chemical name 8-chloro-6, 11-dihydro-11- [1- [ (5-methyl-3-pyridinyl) methyl ] -4-piperidinylidene ] -5H-benzo [5, 6] cyclohepta [1, 2-b ] pyridine fumarate. Rupatadine fumarate is a novel, potent antiallergic agent developed by Uriach pharmaceutical company, Spain, as a selective histamine H1 receptor inverse agonist and a Platelet Activating Factor (PAF) antagonist. In animal experiments, the product shows strong histamine H1 receptor antagonism and PAF receptor blocking effect; has inhibitory effect on mast cell degranulation, neutrophil and eosinophil migration, and cytokine release. The product was first marketed in spain 3/15/2003. Compared with loratadine, cetirizine and the like, the compound has better curative effect and smaller side effect, does not find cardiac toxicity, and can be a first-line medicament for treating allergic rhinitis.

The solubility of rupatadine fumarate in different pH media (CN103108635) is as follows:

pH value of medium	Solubility (g/L)
		1.4	19.60
2.0	7.70
		2.4	5.59
3.1	4.86
		4.1	4.05
5.5	4.06
		6.0	0.96
6.4	0.17
		7.0	0.035

11.0

＜0.005

As can be seen from the above table, rupatadine fumarate is better in solubility under acidic conditions and lower in solubility under neutral conditions, and IF documents of japanese rupatadine fumarate tablets disclose that rupatadine fumarate is extremely insoluble in water and is poor in solubility. Rupatadine fumarate, however, is unstable in acidic solution and forms an adduct of fumaric acid and rupatadine (compound I), as shown in the following formula:

in order to solve the problems of low solubility and stability of rupatadine fumarate, a great deal of research is carried out by those skilled in the art.

CN101669926A discloses a nasal administration liquid preparation of rupatadine fumarate, CN101669901A discloses an ophthalmic administration liquid preparation, and sulfobutyl ether-beta-cyclodextrin, methyl-beta-cyclodextrin and hydroxypropyl-beta-cyclodextrin are added in the two patents to increase the solubility of main drugs. However, both of the above preparations contain cyclodextrin compounds, and long-term administration of the cyclodextrin compound-containing drugs causes damage to children's teeth.

Therefore, how to find a proper dosage form to improve the solubility and stability of rupatadine fumarate still remains a problem to be solved in the technical field.

Disclosure of Invention

The invention aims to overcome the defects in the prior art, and provides the rupatadine fumarate nanoemulsion as well as the preparation method and the application thereof, the solubility of the rupatadine fumarate is increased, the bioavailability is increased, the rupatadine fumarate nanoemulsion is stable in property, does not contain cyclodextrin, and is higher in safety for patients, particularly children patients.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the invention provides rupatadine fumarate nanoemulsion, wherein each 100ml of rupatadine fumarate nanoemulsion comprises the following components:

wherein the appropriate amount of pH regulator is an amount which regulates the pH range of the rupatadine fumarate nano emulsion to 5-6.

The dosage of the auxiliary materials, particularly the dosage of the oil phase, the surfactant and the cosurfactant directly influences the microstructure and the special physicochemical property of the rupatadine fumarate nano-emulsion, and the formed nano-emulsion is unstable even can not form the nano-emulsion when the dosage is improper. When the mass ratio of the surfactant to the cosurfactant is different, the properties and characteristics of the formed nano-emulsion are greatly different, which not only influences the particle size of the nano-emulsion, but also influences the drug loading quality and the drug effect. A large number of experimental researches show that the formed nano emulsion has small particle size, clear and transparent appearance and good fluidity only within the content range of the components.

The rupatadine fumarate nanoemulsion provided by the invention is stable in property and convenient to take, belongs to O/W type nanoemulsion, and has the advantages of average particle size of about 14nm, polydispersity index (PDI) of 0.155 and uniform particle size distribution. The nanoemulsion can increase the solubility of the rupatadine fumarate, can reduce the enzymolysis of the drug in vivo as a drug carrier, can form a protective effect on the drug and improve the absorption of the drug by gastrointestinal tracts, thereby increasing the bioavailability. The rupatadine fumarate nanoemulsion does not contain cyclodextrin, and is higher in safety for patients, particularly children patients.

Preferably, the rupatadine fumarate nanoemulsion comprises the following components per 100 ml:

In any of the above embodiments, preferably, the oil phase comprises at least one of soybean oil, isopropyl myristate, ethyl oleate, dimethicone, and liquid paraffin.

In any of the above embodiments, preferably, the surfactant includes at least one of tween 80, tween 60, polyoxyethylene octylphenol ether-10, span 60, and span 80.

In any of the above embodiments, preferably, the co-surfactant comprises at least one of ethanol, propylene glycol, glycerol, n-butanol, and polyethylene glycol 400.

In any of the above embodiments, preferably, the pH adjuster is one selected from acetic acid-sodium acetate, acetic acid-ammonium acetate, citric acid-disodium hydrogen phosphate, and potassium dihydrogen phosphate-sodium hydroxide.

In any of the above embodiments, preferably, the water includes at least one of ultrapure water, deionized water, and distilled water.

In a second aspect, the present invention provides a method for preparing rupatadine fumarate nanoemulsion according to the first aspect, comprising the following steps:

weighing the cosurfactant and the rupatadine fumarate according to the formula ratio, and uniformly mixing to obtain a first mixture;

weighing the surfactant and the oil phase according to the formula ratio, and uniformly mixing to obtain a second mixture;

adding the first mixture into the second mixture, uniformly mixing, and dropwise adding water while stirring until a stable clear emulsion is formed;

and adding a pH regulator into the stable clear emulsion, regulating the pH to 5-6, and adding water to a constant volume to obtain the rupatadine fumarate nano emulsion.

The preparation method of the rupatadine fumarate nanoemulsion provided by the invention can improve the drug loading capacity to the maximum extent by fractional mixing, is simple and reliable, has good repeatability, does not need special equipment in the preparation process, can be spontaneously formed, and is suitable for industrial production.

In a third aspect, the invention provides an application of the rupatadine fumarate nanoemulsion of the first aspect in preparing an antiallergic drug.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the dosage of the reagent is the dosage of the reagent in the conventional experiment operation if no special description exists; the experimental methods are conventional methods unless otherwise specified.

In a first aspect, an embodiment of the present invention provides rupatadine fumarate nanoemulsion, where each 100ml of the rupatadine fumarate nanoemulsion includes the following components:

The invention discovers that when the pH value is less than 5, the rupatadine fumarate is unstable, the impurity content in the rupatadine fumarate nanoemulsion is higher, and when the pH value is more than 6, the rupatadine fumarate nanoemulsion is unstable, a main drug is easy to separate out, turbidity occurs, and the drug loading is reduced. Only the pH is limited within the range of 5-6, the rupatadine fumarate nanoemulsion is most stable and the impurity content is lowest.

The rupatadine fumarate nanoemulsion provided by the invention is stable in property and convenient to take, belongs to O/W type nanoemulsion, has an average particle size of about 14nm, a polydispersity index (PDI) of 0.155, and uniform particle size distribution, and is small in particle size, large in drug loading capacity and high in stability. The nanoemulsion can increase the solubility of the rupatadine fumarate, can reduce the enzymolysis of the drug in vivo as a drug carrier, can form a protective effect on the drug and improve the absorption of the drug by gastrointestinal tracts, thereby increasing the bioavailability. The rupatadine fumarate nanoemulsion does not contain cyclodextrin compounds, and has higher safety for patients, particularly children patients.

Further, each 100ml of the rupatadine fumarate nanoemulsion comprises the following components:

Further, the oil phase comprises at least one of soybean oil, isopropyl myristate, ethyl oleate, dimethyl silicone oil and liquid paraffin. The selection of the oil phase is crucial to the solubilization of the drug and the existence of a single-phase region of the nanoemulsion, the shorter the hydrocarbon chain of the oil, the deeper the organic phase penetrates into an interfacial film, and the more stable the nanoemulsion is, but the oil phase with the longer hydrocarbon chain is helpful for increasing the dissolution of the drug, so the contradiction is overcome by combining the comprehensive consideration of the dissolution condition of the rupatadine fumarate. The oil phase in the embodiment can be selected to increase the solubility of the rupatadine fumarate, and the formed nanoemulsion is relatively stable. Preferably, the oil phase is ethyl oleate, the taste of the rupatadine fumarate nanoemulsion can be improved by selecting the ethyl oleate as the oil phase, and the safety of the ethyl oleate is higher.

Further, the surfactant comprises at least one of tween 80, tween 60, polyoxyethylene octyl phenol ether-10, span 60 and span 80. The type and amount of the surfactant directly influence whether the nanoemulsion can be formed and the toxicity. The surfactant in the embodiment can reduce the oil-water interfacial tension, form a firm emulsion film and has a solubilizing effect on the rupatadine fumarate. Preferably, the surfactant is a mixture of tween 80 and span 80, wherein the mass ratio of tween 80 to span 80 is 2-4, for example, the mass ratio of tween 80 to span 80 can be 2, 3 or 4.

Further, the cosurfactant comprises at least one of ethanol, propylene glycol, glycerol, n-butanol and polyethylene glycol 400, preferably, the cosurfactant is propylene glycol, the propylene glycol can obviously improve the solubility of the rupatadine fumarate, the drug loading rate is increased, the particle size of the nanoemulsion is reduced through self shearing action, and the nanoemulsion area is the largest when Km (mass ratio of the surfactant to the cosurfactant) is 2: 1.

Further, the pH regulator is one selected from acetic acid-sodium acetate, acetic acid-ammonium acetate, citric acid-disodium hydrogen phosphate, and potassium dihydrogen phosphate-sodium hydroxide.

Further, the water includes at least one of ultrapure water, deionized water, and distilled water.

In a second aspect, an embodiment of the present invention provides a method for preparing rupatadine fumarate nanoemulsion according to the first aspect, including the following steps:

In a third aspect, the embodiment of the invention provides an application of the rupatadine fumarate nanoemulsion of the first aspect in preparing an antiallergic drug.

The invention is described in further detail with reference to a part of the test results, which are described in detail below with reference to specific examples.

The propylene glycol described in the following examples and comparative examples is 1, 2-propanediol.

The component contents of rupatadine fumarate nanoemulsion of examples 1-3 and comparative examples 1-2 are shown in table 1, calculated on 100ml rupatadine fumarate nanoemulsion.

TABLE 1 examples 1-3 and comparative examples 1-2 Lapatadine fumarate nanoemulsion component content

Example 1

The components of rupatadine fumarate nanoemulsion provided in this example are shown in table 1.

The preparation method of the rupatadine fumarate nanoemulsion provided by the embodiment comprises the following steps:

(1) weighing 10g of propylene glycol and 128mg of rupatadine fumarate, mixing and uniformly stirring to obtain a first mixture;

(2) weighing 15g of tween 80, 5g of span 80 and 50g of ethyl oleate, mixing and uniformly stirring to obtain a second mixture;

(3) adding the first mixture into the second mixture, mixing and stirring uniformly, and dropwise adding distilled water while stirring until a stable clear transparent emulsion is formed;

(4) adding 0.397g of citric acid and 0.568g of disodium hydrogen phosphate into the stable clear transparent emulsion, adjusting the pH to 5.2, adding distilled water to a constant volume of 100mL, and filtering to obtain the rupatadine fumarate nanoemulsion.

Example 2

(2) weighing 20g of tween 80, 5g of span 80 and 45g of ethyl oleate, mixing and uniformly stirring to obtain a second mixture;

(4) adding 0.320g of citric acid and 0.568g of disodium hydrogen phosphate into the stable clear transparent emulsion, adjusting the pH to 5.5, adding distilled water to a constant volume of 100mL, and filtering to obtain the rupatadine fumarate nanoemulsion.

Example 3

(1) weighing 7g of propylene glycol and 128mg of rupatadine fumarate, mixing and uniformly stirring to obtain a first mixture;

(2) weighing 10g of tween 80, 5g of span 80 and 55g of ethyl oleate, mixing and uniformly stirring to obtain a second mixture;

(4) adding 0.246g of citric acid and 0.568g of disodium hydrogen phosphate into the stable clear transparent emulsion, adjusting the pH to 6.0, adding distilled water to a constant volume of 100mL, and filtering to obtain the rupatadine fumarate nanoemulsion.

Comparative example 1

The components of the rupatadine fumarate nanoemulsion provided by the comparative example are shown in table 1.

The preparation method of the rupatadine fumarate nanoemulsion provided by the comparative example comprises the following steps:

(2) weighing 15g of tween 80, 5g of span 80 and 30g of ethyl oleate, mixing and uniformly stirring to obtain a second mixture;

Comparative example 2

(1) weighing 30g of propylene glycol and 128mg of rupatadine fumarate, mixing and uniformly stirring to obtain a first mixture;

(2) weighing 15g of tween 80, 5g of span 80 and 20g of ethyl oleate, mixing and uniformly stirring to obtain a second mixture;

Example 4

(1) Investigation of centrifugal stability

This example examines the centrifugal stability of rupatadine fumarate nanoemulsions prepared in examples 1-3 and comparative examples 1-2. Under the condition of room temperature, the rupatadine fumarate nano-emulsion prepared in the examples 1-3 and the comparative examples 1-2 is placed in a centrifuge tube, centrifuged for 30min at 4000 rpm and 14000 rpm on a centrifuge respectively, whether the emulsion is layered or not is observed, and the test result is shown in table 2.

TABLE 2 centrifugation results of rupatadine fumarate nanoemulsions prepared in examples 1-3 and comparative examples 1-2

As can be seen from Table 2, the nano-emulsions prepared in examples 1 to 3 did not undergo delamination even at high rotation speeds, and the centrifugal stability was very good, while comparative examples 1 to 2 all underwent delamination at high rotation speeds. The invention finds that the stability of the nano emulsion is poor although clear and stable nano emulsion can be obtained after changing the contents of the oil phase and the cosurfactant.

(2) Temperature stability investigation

The rupatadine fumarate nanoemulsion prepared in examples 1-3 and comparative examples 1-2 was filled in a transparent colorless glass bottle, sealed, placed at two temperature conditions of room temperature (25 ℃) and 40 ℃ for 40 days, sampled every 5 days to observe the properties, and the results are shown in tables 3-4.

TABLE 325 ℃ retention of the sample for observation

TABLE 440 ℃ sample observation

As can be seen from tables 3 and 4: the nanoemulsion of the embodiment of the invention is kept clear and transparent through a temperature stability test, and has no unstable demulsification phenomena such as layering, turbidity or crystal precipitation, so that the nanoemulsion is a stable pharmaceutical dosage form. On the contrary, the nanoemulsion obtained by adopting the formula of the comparative example has unstable demulsification phenomenon, turbid solution and unstable system.

Example 5

This example examines the long-term stability of rupatadine fumarate nanoemulsions prepared in examples 1-3 and comparative examples 1-2. The prepared rupatadine fumarate nanoemulsion is sealed in a brown glass bottle, is placed at room temperature (25 ℃) and relative humidity (60 +/-5)% for 12 months, is sampled at 0, 3, 6, 9 and 12 months respectively, the characters of the nanoemulsion and the content of the main drug are inspected, the result is shown in table 5, the determination method of the content of the main drug is high performance liquid chromatography (0512 of the four ministry of China pharmacopoeia 2020 edition), and the specific operation steps are as follows:

the solvent is methanol-water (volume ratio 50: 50).

Preparing an ammonium acetate buffer salt solution: 1.54g of ammonium acetate is taken, dissolved in 1000mL of water and the pH is adjusted to 6.0 +/-0.05 by dilute acetic acid solution.

Preparing a test solution: precisely measuring 5mL of rupatadine fumarate nanoemulsion prepared in examples 1-3 and comparative examples 1-2, putting the rupatadine fumarate nanoemulsion into a 50mL measuring flask, adding a proper amount of solvent, shaking to dissolve, diluting with the solvent to a scale, and shaking up.

Preparing a reference substance solution: precisely weighing a proper amount of rupatadine fumarate reference substance (about equal to 10mg of rupatadine), putting the reference substance into a 10mL measuring flask, adding a proper amount of solvent, carrying out ultrasonic treatment to dissolve the reference substance, diluting the reference substance to a scale by using the solvent, shaking the reference substance uniformly, precisely weighing 1mL, putting the reference substance into the 10mL measuring flask, diluting the reference substance to the scale by using the solvent, and shaking the reference substance uniformly.

Chromatographic conditions are as follows: octyl silane bonded silica gel is used as a filling agent, ammonium acetate buffer salt solution-methanol (volume ratio is 25: 75) is used as a mobile phase, the flow rate is 1.0mL per minute, the column temperature is 40 ℃, the detection wavelength is 264nm, and the injection volume is 20 mu l.

System applicability requirements: in the chromatogram of the control solution, the number of theoretical plates is not less than 3000 calculated according to the rupatadine peak.

The content determination of the main drug of the test sample: precisely measuring the test solution and the reference solution, respectively injecting into a high performance liquid chromatograph, and recording the chromatogram. Calculating C in the sample by peak area according to external standard method₂₆H₂₆ClN₃The content of (a).

Quality standard: the product contains rupatadine (C)₂₆H₂₆ClN₃) Should be 95.0% -105.0% of the marked amount.

The labeled amount of the rupatadine fumarate nanoemulsion prepared in examples 1-3 and comparative examples 1-2 was 1 mg/mL.

The content of main drugs: the percentage of the quantity is indicated.

TABLE 5 Long-term stability test results of rupatadine fumarate nanoemulsion prepared in examples 1-3 and comparative examples 1-2

The results in table 5 show that the nanoemulsion of the embodiment of the invention maintains clear and transparent after long-term stability tests, is a stable drug formulation, has no obvious change in the content of the main drug, and has good stability.

Example 6

In this example, the safety of rupatadine fumarate nanoemulsion prepared in examples 1-3 was examined. 120 rats were divided into A, B groups and C groups, each group consisting of 40 rats, and the rupatadine fumarate nanoemulsion prepared in example 1 was administered to the A group of rats, the rupatadine fumarate nanoemulsion prepared in example 2 was administered to the B group of rats, and the rupatadine fumarate nanoemulsion prepared in example 3 was administered to the C group of rats, both in clinical dose, for 30 days by continuous gavage. In the test period, the animals of each test group grow well, and indexes such as body weight, food intake, blood routine, blood biochemistry, organ coefficient and the like are all in normal ranges, which shows that the rupatadine fumarate nano-emulsion prepared in examples 1-3 is good in safety.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims

1. The rupatadine fumarate nanoemulsion is characterized in that each 100 milliliters of the rupatadine fumarate nanoemulsion comprises the following components:

2. The rupatadine fumarate nanoemulsion of claim 1, wherein the rupatadine fumarate nanoemulsion comprises the following components per 100 milliliters:

3. The rupatadine fumarate nanoemulsion of claim 1, wherein the oil phase comprises at least one of soybean oil, isopropyl myristate, ethyl oleate, dimethicone, liquid paraffin.

4. The rupatadine fumarate nanoemulsion of claim 1, wherein the surfactant comprises at least one of tween 80, tween 60, polyoxyethylene octylphenol ether-10, span 60, and span 80.

5. The rupatadine fumarate nanoemulsion of claim 1, wherein the co-surfactant comprises at least one of ethanol, propylene glycol, glycerol, n-butanol, polyethylene glycol 400.

6. The rupatadine fumarate nanoemulsion of claim 1, wherein the pH adjuster is selected from one of acetic acid-sodium acetate, acetic acid-ammonium acetate, citric acid-disodium hydrogen phosphate, and potassium dihydrogen phosphate-sodium hydroxide.

7. The rupatadine fumarate nanoemulsion of claim 1, wherein the water comprises at least one of ultrapure water, deionized water, distilled water.

8. A process for the preparation of rupatadine fumarate nanoemulsion of any of claims 1-7, comprising the following steps:

9. Use of rupatadine fumarate nanoemulsion of any one of claims 1-7 for the preparation of an antiallergic drug.