CN100428937C

CN100428937C - Levosimendan freeze-drying composition

Info

Publication number: CN100428937C
Application number: CNB2003101022724A
Authority: CN
Inventors: 王晋; 妥福军
Original assignee: HONGSHENG MEDICINE DEVELOPMENT Co Ltd XI'AN
Current assignee: HONGSHENG MEDICINE DEVELOPMENT Co Ltd XI'AN
Priority date: 2003-10-30
Filing date: 2003-10-30
Publication date: 2008-10-29
Anticipated expiration: 2023-10-30
Also published as: CN1611220A

Abstract

The present invention relates to a medicinal composition comprising levosimendan as an active ingredient and a preparation method thereof, particularly to a powder injection comprising levosimendan as an active ingredient and a preparation method thereof. Compared with the existing congener injection, the powder injection of the present invention has the advantages of high stability, long storage period, and convenient transportation, storage and use. The composition also contains citric acid, phospholipid, cyclodextrin and derivatives thereof or a surface active agent. The product has favorable solubility and stability.

Description

Levosimendan freeze-dried composition

Technical Field

The levosimendan powder of the present invention is pharmaceutical and the main route of administration is intravenous injection. The injectable powders of the invention have good stability and they are particularly suitable for infusion. Levosimendan, chemical name (R) - (-) -2- [4(4 methyl-6 oxo-1, 4, 5, 6-tetrahydropyridazin-3) -yl-phenylhydrazono ] malononitrile, for clinical use in the treatment of congestive heart failure.

Background of the invention

Levosimendan, the levorotatory form of (-) -2- [ 4- (4-methyl-6-oxo-1, 4, 5, 6-tetrahydropyridazin-3) -yl-phenylhydrazono ] malononitrile, is disclosed in EP 565546B 1 and WO 97/35841. Levosimendan is effective in the treatment of heart failure and exhibits a marked dependence on binding to troponin. The structural formula of levosimendan is as follows:

the blood flow effects of levosimendan in humans are disclosed by Sundberg, s. et al in am.j.cardiol.1995; 75: 1061 to 1066. The pharmacokinetics of levosimendan after intravenous and oral administration is disclosed by Sandell E-p, et al in j.cardiovasc, pharmacol, 26(suppl.1), S57-S62, 1995 levosimendan for the treatment of myocardial ischemia is disclosed in WO 93/21921. Formulations for transdermal administration of levosimendan are disclosed in WO 98/01111. Clinical studies have demonstrated the beneficial effects of levosimendan on patients with heart failure.

Administration by the vascular route, for example, intravenous injection, has many advantages, including, inter alia: (1) intravenous injection or infusion can achieve a response almost immediately; (2) the therapeutic response is more easily controlled by vascular administration; (3) patients who cannot take drugs orally due to loss of consciousness may be dosed via blood vessels, or may have impaired absorption due to inactivation of the gastrointestinal tract.

The preparation of solutions for levosimendan injection is disclosed in WO01/19334, and injection solutions, in particular suitable for intravenous route, involve many problems of levosimendan stability due to chemical and physical factors. Levosimendan solutions are more sensitive to chemical factors, which limits the shelf life of the solution and produces unwanted degradation products. Levosimendan is sensitive to heat, the storage temperature of the levosimendan injection on the market in Europe is 2-8 ℃, the color of the injection can be deepened from yellow green to orange along with the prolonging of the storage time, the content is reduced, and related substances are increased. Levosimendan is poorly soluble in water and readily precipitates from aqueous solutions. Once precipitated, solutions for intravenous injection are dangerous because particles may block blood vessels. The solubility of levosimendan decreases rapidly as the pH decreases from neutral, and therefore, a low pH is in principle detrimental to solubility. Thus, there is a need for improved aqueous levosimendan formulations which are stable to chemical and physical factors during long-term storage and which are suitable for intravenous administration.

Therefore, there is a great need in the art for levosimendan formulated to have good solubility and stability. The entire text of the patent specification

Through a great deal of research on the prescription and the preparation process, the invention is completed through further research on the basis that the problems can be effectively solved by adopting the freeze drying process.

The conventional powder injection can solve the problem of the stability of the preparation, but the dissolution performance of levosimendan in the product is not obviously improved, other excipients are also required to be added, besides a pH regulator, namely organic acid, is added to ensure the stability of the levosimendan, phospholipid, poloxamer, polyethylene glycol, beta-cyclodextrin and derivatives or surfactants thereof are also required to be added, a stable freeze-dried product can be prepared by freeze drying, and the stable freeze-dried product can be quickly re-dispersed into a clear colloidal dispersion or solution by adding an aqueous diluent for medicine when in use. In addition, the composition does not contain organic solvents such as ethanol and the like, so that irritation reaction is avoided.

It is an object of the present invention to provide lyophilized compositions of levosimendan containing phospholipids which are amphiphilic and which make them particularly suitable for formulating levosimendan which is less stable in the solution state and poorly water soluble in the present invention. This is very beneficial for extending the shelf life and improving the solubility of levosimendan formulations. The liposome or lipid complex containing levosimendan can be prepared by adopting a freeze drying method, a film drying method and a reverse phase evaporation method which are commonly used in the field and adjusting the dosage of phospholipid, then stable freeze-dried powder is prepared by freeze drying, and the obtained freeze-dried product can be quickly re-dispersed into colloidal dispersion after being diluted by a proper amount of medicinal aqueous diluent. The phospholipids used in the present invention are not limited to a particular class of phospholipids, including natural or synthetic phospholipids, and exemplary phospholipids include, but are not limited to, lecithin, soybean phospholipid, cephalin, phosphatidic acid, phosphatidylcholine dipalmitoyl, phosphatidylethanolamine, diaminocholesterol, soybean steroyl glycoside, soybean sterol, ergosterol, and mixtures thereof. Advantageously, the weight ratio of levosimendan to phospholipid is from about 1: 1 to about 1: 100, preferably from about 1: 5 to about 1: 25.

Levosimendan is chemically stable at a pH in the range of 3.0 to 4.5 and therefore the pH can be adjusted to this range by the addition of organic acids, preferred acidic compounds include pharmaceutically acceptable organic acids having a pKa in the range of 2 to 4, including 2-hydroxyalkanoic acids such as citric, lactic, tartaric or malic acid. If a pharmaceutically acceptable buffer system is used, the buffer is selected from the class of buffers that are effective in maintaining the pH below 5.5, preferably below 5 or less, and most preferably in the range of 3 to 4.5, and the buffer is formulated using well-known methods.

It is another object of the present invention to provide a lyophilized levosimendan mixture containing cyclodextrin. Inclusion of the poorly soluble drug levosimendan with cyclodextrin significantly improves its physicochemical properties, for example: the drug can be powdered to improve the stability and solubility, preferably, the cyclodextrin and the derivative thereof are selected from beta-cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin, hydroxyethyl-beta-cyclodextrin and a mixture thereof, preferably 2-hydroxypropyl-beta-cyclodextrin. Wherein the molar ratio of levosimendan to cyclodextrin is about 1: 1 to 1: 20, preferably 1: 1 to 1: 5. The conditions of inclusion temperature, dispersion force, stirring speed and the like can be designed and selected by adopting a conventional method.

It is another object of the present invention to provide a lyophilized levosimendan composition containing a surfactant, which is a risk for clinical use, in which the lyophilized levosimendan formulation containing levosimendan is often back-turbid or crystallized after dilution with a pharmaceutically acceptable diluent as described above. The present invention facilitates rapid dissolution of levosimendan by solubilizing it using a surfactant (e.g., poloxamer) while avoiding the use of irritating solvents. Studies show that the povidone, the polyethylene glycol, the poloxamer, the polyoxyethylene castor oil, the polyoxyethylene hydrogenated castor oil, the Tween-20, the Tween 80 or the mixed system thereof. The amount of surfactant used can be determined by routine experimentation to ensure a clear solution after redispersion. Advantageously, the weight ratio of levosimendan to surfactant is from about 1: 1 to about 1: 200, preferably from about 1: 5 to about 1: 50.

Preferably, a pharmaceutical freeze-drying excipient can be further added into the pharmaceutical composition. Preferably, the excipient is selected from the group consisting of mannitol, sorbitol, sodium chloride, glucose, fructose, xylitol, lactose, and mixtures thereof. Mannitol is preferred. The weight ratio of levosimendan to the excipient is 1: 50-1: 1000, preferably 1: 100-1: 300.

The active substance levosimendan is used in an amount of about 2.5mg to about 25mg, preferably about 5mg to about 12.5mg, based on the total weight of the composition.

The freeze-drying period can be adjusted by those skilled in the art according to the requirements of clinical preparations and specific production equipment, and generally, the uniform preparation can be placed in a vial of 5-50 ml, the pre-freezing temperature is-20 ℃ to-60 ℃, preferably-30 ℃ to 40 ℃, and the pre-freezing time is 1 hour to 6 hours, preferably 3-4 hours. The freeze-drying temperature is-10 ℃ to-30 ℃, preferably-20 ℃ to-25 ℃, and the freeze-drying time is 10 hours to 40 hours, preferably 20 hours to 24 hours. The final moisture content of the lyophilized composition is generally less than 5%, preferably 1% to 1.5%.

These lyophilised compositions of the invention are suitable for clinical use. Before use, a suitable amount of a sterile pharmaceutically acceptable aqueous diluent (e.g., water for injection, physiological saline, aqueous dextrose, xylitol, and other known aqueous carriers) may be added and redispersed into a colloidal dispersion or solution formulation for intramuscular injection or intravenous drip (intravenous administration).

The following examples are intended to further illustrate the present invention and are not intended to limit the scope of the present invention.

Example 1: preparing powder injection containing phospholipid

Taking 12.5mg of levosimendan, 10mg of citric acid, 75mg of phospholipid and 10mg of cholesterol, and adding 5ml of absolute ethyl alcohol. Stirring to completely dissolve, drying under reduced pressure on a rotary evaporator, removing solvent, adding 5ml of water, stirring to be uniform, performing ultrasonic oscillation, filtering, adding 300mg of mannitol, placing in a sterile vial, performing freeze drying in a freeze dryer, and sealing to obtain light yellow-light yellow levosimendan powder injection.

Example 2: preparing powder injection containing cyclodextrin

Taking 12.5mg of levosimendan and 10mg of citric acid, using 5ml of absolute ethyl alcohol, stirring until the levosimendan and the citric acid are completely dissolved, taking 150mg of 2-hydroxypropyl-beta-cyclodextrin, adding 25ml of water, adding the levosimendan solution into the 2-hydroxypropyl-beta-cyclodextrin solution under stirring, and preparing the uniform cyclodextrin inclusion compound. Drying under reduced pressure on a rotary evaporator, and volatilizing ethanol. Adding mannitol 300mg under stirring, filtering the solution with sterilized sintered glass funnel, placing the filtrate in sterile penicillin bottle, freeze drying in freeze dryer, and sealing. Thus obtaining the light yellow-light yellow levosimendan powder injection.

The pharmaceutical composition prepared in example 3 was confirmed to be in the form of inclusion compound by X-ray powder diffraction and thermal analysis (DSC).

Example 3: preparing powder injection containing surfactant

Taking 12.5mg of levosimendan, 50.0mg of polyethylene glycol, 150mg of polyoxyethylene castor oil and 0.3g of mannitol, adding 5ml of water, and stirring until the levosimendan, the polyethylene glycol, the polyoxyethylene castor oil and the mannitol are completely dissolved. Filtering with 0.22 μm microporous membrane, placing in sterile penicillin bottle to obtain yellowish clear solution, freeze drying in a freeze dryer, and sealing. Thus obtaining the light yellow-light yellow levosimendan powder injection.

The quality and stability of levosimendan powder injection and the animal anaphylaxis, hemolysis and vascular irritation are researched.

1. Content determination:

high performance liquid chromatography: octadecylsilane chemically bonded silica is used as a filling agent; dissolving in phosphate buffer (sodium dihydrogen phosphate 2.34g, adding water 950ml, adding 5% phosphoric acid solution about 3.7ml, adjusting pH to 3.0 + -0.1, adding water to 1000ml) -acetonitrile (60: 40) as mobile phase; the flow rate was 1ml per minute; the detection wavelength was 360 nm.

Assay samples were formulated to give levosimendan concentrations of 75 μ g/ml, a procedure which was not specifically described. Injecting 20 mu l of the mixture into a liquid chromatograph, and recording a chromatogram; another reference levosimendan (7.5 mg) is weighed precisely and placed in a 100ml measuring flask, 10ml of methanol is added to dissolve the reference levosimendan, the reference levosimendan is diluted to the scale with a mobile phase and shaken up, and the measurement is carried out by the same method. Calculating according to the peak area by an external standard method to obtain the product.

2. Related substances

The sample is prepared into a sample solution and a control solution which respectively contain 200 mu g and 2 mu g of levosimendan in each 1ml, and the specific preparation method is omitted. According to the method of content determination, 20 μ l of the control solution is injected into a liquid chromatograph, the detection sensitivity is adjusted to make the peak height of the main component about 20% of the full range, then 20 μ l of each of the test solution and the control solution is injected into the liquid chromatograph, and the chromatogram is recorded until the retention time of the main component peak is 6 times. The peak area of the impurity, except the solvent peak, shown in the chromatogram of the test solution should not be more than 1.5 times (1.5%) of the area of the main bee of the control solution.

Comparing the stability of powder injection and injection

The stability of levosimendan powder injection prepared in example 3 and conventional small water injection is examined, the influencing factors are determined by placing the levosimendan powder injection in 40 ℃ +/-2 ℃ for 10 days under the illumination of 4500LX +/-500 LX, sampling is carried out on the 5 th day and the 10 th day respectively, and the examined data are shown in the following table 1.

TABLE 1 test for influence factors to examine the thermal and light stability of powder and solution

The above results show that: the powder injection can be quickly redispersed into clear liquid after being added with water; under the same experimental conditions, the stability of the powder injection is superior to that of a solution, and particularly the appearance of the powder injection is far superior to that of the solution. The good solubility and stability not only ensure the safety of clinical medication, but also correspondingly prolong the effective period of levosimendan.

4. Allergy, hemolytic and vascular irritation studies

The levosimendan powder injection of the invention has no anaphylaxis (guinea pig test), hemolytic property and vascular irritation (big ear white rabbit test).

Claims

1. A freeze-dried pharmaceutical composition comprises the following three components:

i. the amount of levosimendan to be administered,

(ii) an organic acid, wherein,

a phospholipid, or, a beta cyclodextrin and derivatives thereof,

wherein the organic acid is one or a mixture of citric acid, lactic acid, tartaric acid or malic acid; the phospholipid is selected from one or more of lecithin, soybean phospholipid, cephalin, phosphatidic acid, dipalmitoyl phosphatidylcholine, hydrogen phosphatidyl ethanolamine and phosphatidyl serine; the beta cyclodextrin and the derivatives thereof are selected from one or a mixture of more of beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin and hydroxyethyl-beta-cyclodextrin; and the pharmaceutical composition further comprises a pharmaceutically acceptable freeze-drying excipient, wherein the freeze-drying excipient is selected from one or a mixture of mannitol, sorbitol, sodium chloride, glucose, fructose, sucrose, xylitol and lactose, and the pharmaceutical composition does not contain ethanol.

2. The pharmaceutical composition according to claim 1 wherein the weight ratio of levosimendan to the organic acid is from 1: 0.1 to 1: 10.

3. The pharmaceutical composition according to claim 1 wherein the weight ratio of levosimendan to said mixture of one or more phospholipids is from 1: 1 to 1: 75.

4. The pharmaceutical composition of claim 1 wherein the weight ratio of levosimendan to beta-cyclodextrin and its derivatives is 1: 1 to 1: 15.

5. The pharmaceutical composition of claim 1 wherein the weight ratio of levosimendan to the freeze-dried excipient is from 1: 50 to 1: 1000.

6. The pharmaceutical composition of claim 5 wherein the weight ratio of levosimendan to the freeze-dried excipient is from 1: 100 to 1: 300.

7. A process for preparing a pharmaceutical composition according to any one of claims 1 to 6 by one of the following methods:

the method comprises the following steps: dissolving the pharmaceutical composition in water, optionally filtering for sterilization, directly freeze-drying, and then subpackaging freeze-dried powder; or,

the method 2 comprises the following steps: dissolving the pharmaceutical composition in water, then subpackaging the obtained solution in 5ml, 10ml, 20ml or 50ml of freeze-drying vials, placing the vials into metal trays with the sizes corresponding to those of freeze-drying boxes without sealing, sterilizing the freeze-drying boxes, reducing the temperature of a partition plate of the freeze-drying boxes to-30 ℃ to-40 ℃ and maintaining the temperature for 1-2 hours, then carrying out vacuum treatment on the freeze-drying boxes to ensure that the vacuum degree in the freeze-drying boxes is 0.2-0.1 Pa, then heating the partition plate, strictly controlling the temperature of the partition plate to be 5 ℃ below the melting point, keeping the temperature for 12-24 hours, finally heating to 25-30 ℃, keeping the temperature for a certain time to ensure that the temperature of the preparation is coincided with the temperature of the partition plate, placing sterile filtered gas, controlling the water content of the preparation to be 0.1-3%, and rapidly sealing.

8. Use of a pharmaceutical composition according to any one of claims 1 to 6 for the manufacture of a medicament for the short-term treatment of adults with weakened myocardial contractility due to organic heart disease or after cardiac surgery.