JP7582205B2 - A stable lyophilized formulation containing daptomycin - Google Patents

Description

The present invention relates to a stable lyophilized formulation containing daptomycin.

Daptomycin is a cyclic lipopeptide antibiotic that is used to treat sepsis, infective endocarditis, deep skin infections, secondary infections in trauma, burns, and surgical wounds, and secondary infections in erosions and ulcers. Daptomycin is prescribed as an intravenous solution to treat infections caused by several gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) (Patent Document 1, etc.). Daptomycin for injection (Cubicin (registered trademark), MSD K.K.) is provided as a lyophilized powder (Non-Patent Document 1, etc.).
However, storage in a cool place, usually at 2°C to 8°C, is required.

Special Publication No. 2013-511557

Package insert for Cubicin 350mg for intravenous injection

Under these circumstances, there is a need for a formulation containing daptomycin that is stable when stored at room temperature.
An object of the present invention is to provide a freeze-dried preparation containing daptomycin that can maintain stability when stored at room temperature.

As a result of extensive research, the present inventors have found that the use of a specific stabilizer can stabilize a lyophilized preparation containing daptomycin to such an extent that it can be stored at room temperature.
This application includes the following inventions:
[1] daptomycin,
A stable freeze-dried preparation comprising one or more stabilizers selected from L-arginine, inositol, dextran, meglumine, macrogol and L-histidine.
[2] The freeze-dried preparation described above, wherein the stabilizer is one or more selected from the group consisting of L-arginine, inositol and dextran.
[3] The freeze-dried preparation described above, wherein the stabilizer is L-arginine.
[4] The stable freeze-dried preparation according to the above, wherein the pH of the preparation when dissolved with water for injection is 5.5 to 7.5.

According to the present invention, a freeze-dried formulation containing daptomycin can be provided that can maintain stability when stored at room temperature.

1 is a graph showing the increase in total related substances relative to the amount of L-arginine added in a daptomycin-containing lyophilized formulation of the present invention.

The daptomycin-containing lyophilized formulation of the present application is a lyophilized formulation that can maintain the stability of daptomycin even when stored at room temperature, i.e., a solid daptomycin-containing formulation for preparing a liquid formulation containing daptomycin.
Daptomycin is disclosed, for example, in US Pat. No. 4,537,717 and is a cyclic polypeptide antibiotic having the chemical formula C ₇₂ H ₁₀₁ N ₁₇ O ₂₆ and a molecular weight of 1620.67.
The daptomycin-containing lyophilized preparation of the present application contains one or more stabilizers selected from L-arginine, inositol, dextran, meglumine, macrogol, and L-histidine.
The content of the stabilizer may be 3% by mass to 250% by mass, preferably 6% by mass to 200% by mass, and more preferably 10% by mass to 136% by mass, based on the total mass of daptomycin. From another viewpoint, the content may be 1% by mass to 80% by mass, preferably 5% by mass to 70% by mass, and more preferably 10% by mass to 60% by mass, based on the total mass of the lyophilized formulation. Specifically, the content is preferably 10 mg to 800 mg, more preferably 20 mg to 600 mg, even more preferably 30 mg to 500 mg, and particularly preferably 35 mg to 500 mg, based on 367.5 mg of daptomycin.
In particular, when L-arginine is contained, the amount is preferably 200 mg to 800 mg, more preferably 300 mg to 600 mg, and even more preferably 350 mg to 500 mg.
When inositol is contained, the amount is preferably 1 mg to 150 mg, more preferably 10 mg to 100 mg, and even more preferably 20 mg to 80 mg.
When dextran is contained, the amount is preferably 10 mg to 500 mg, more preferably 50 mg to 300 mg, and even more preferably 70 mg to 250 mg. The type of dextran is not particularly limited, but dextran 40 is preferred.
When meglumine is contained, the amount is preferably 10 mg to 400 mg, more preferably 30 mg to 200 mg, and even more preferably 50 mg to 150 mg.
When macrogol is contained, the amount is preferably 10 mg to 500 mg, more preferably 50 mg to 300 mg, and even more preferably 70 mg to 250 mg. The type of macrogol is not particularly limited, but macrogol 4000 is preferred.
When L-histidine is contained, the amount is preferably 10 mg to 500 mg, more preferably 50 mg to 300 mg, and even more preferably 70 mg to 250 mg.

In addition to the stabilizers described above, the composition may further contain stabilizers known in the art. Examples of such stabilizers include those exemplified in JP 2004-10511 A, JP 05-306235 A, and JP 2001-525372 A. The amount of stabilizer other than the stabilizers described above may be 0.1% by mass to 100% by mass, preferably 0.1% by mass to 99.99% by mass, based on the total mass of daptomycin.

The daptomycin-containing lyophilized formulation of the present application may contain any additives commonly used in pharmaceuticals, as necessary, including additives known in the art, such as pH adjusters, isotonicity agents, bulking agents, antioxidants, preservatives/antiseptics, carbohydrates, water-soluble polymers, hydrophilic or hydrophobic materials, gelatin, oils, etc.
Examples of preservatives/antiseptics include cresol, benzyl alcohol, phenol, benzalkonium chloride, benzethonium chloride, chlorobutanol, phenylethyl alcohol, methylparaben, propylparaben, thimerosal, nitric acid, and phenylmercuric acetate, but it is preferable that they are not included.

The daptomycin-containing lyophilized formulation of the present application can be produced by preparing a solution containing daptomycin, one or more stabilizers selected from L-arginine, inositol, dextran, meglumine, macrogol, and L-histidine, and optionally additives, and filtering the solution through sterile filtration, filling the solution into vials, and lyophilizing the solution according to standard methods.
The solution may be prepared by a method known in the art. For example, daptomycin or the like is dissolved in an aqueous medium at room temperature or with cooling. Dissolution may be performed manually or with a stirrer or the like. The solution may be prepared using a metal container or a non-metallic container such as glass or plastic. Examples of the aqueous medium include water for injection, distilled water for injection, physiological saline, buffer solution, and replacement fluid (infusion such as electrolyte infusion), with distilled water for injection being preferred.
From the viewpoint of drug stability, the pH of the obtained solution is preferably 5.5 to 7.5, and more preferably 6.0 to 7.0. The pH can be adjusted using an acid such as hydrochloric acid or an alkali such as sodium hydroxide. A pharma- ceutically acceptable buffer solution may also be used.
The aqueous medium may be used in an amount of, for example, 2 ml to 6 ml relative to the total mass of daptomycin, or from another perspective, the concentration of daptomycin may be 50 w/v % to 275 w/v %.
It is preferable to dissolve daptomycin and the like while cooling the aqueous medium. The cooling temperature can be, for example, 20° C. or lower, and preferably 15° C. or lower.
When a freeze-dryer is used in the next step as described below, the cooling may be carried out during transfer to the freeze-dryer and/or within the dryer.
Sterile filtration can be performed, for example, using a filter. The resulting solution may be filtered through a filter with a pore size of 0.2 μm to 0.45 μm, for example, a pore size of 0.22 μm. The filter may be, for example, a cartridge filter made of polyvinylidene fluoride. Sterilization by filtration includes any process that removes or kills all microbial life forms, including infectious agents (fungi, bacteria, viruses, spore forms, etc.) contained in a liquid. Two or more filters with different pore sizes may be used. Typically, a filter with a pore size of 0.22 μm can remove microorganisms as a microfiltration. Sterilization can be performed by methods known in the art. Sterilization can utilize heat, chemicals, radiation, high pressure, high pressure steam, etc., but it is preferable to adopt a sterilization method that does not involve heat load.

The lyophilization of the cooled daptomycin-containing solution can be carried out under commonly applied conditions by using known lyophilization methods such as tray lyophilization, spray lyophilization, and vial/syringe lyophilization. For this purpose, it is preferable to use a lyophilizer capable of carrying out these lyophilization methods. In lyophilization, it is usually preferable to dry the lyophilized product so that its moisture content is 3.0% by mass or less, more preferably 2.0% by mass or less, and even more preferably 1.5% by mass or less.

The lyophilized preparation is preferably a preparation filled in a container such as a vial or a syringe. In particular, from the viewpoint of easy operation during use, the lyophilized preparation is preferably in the form of, for example, a double-chamber or single-chamber pre-filled syringe. In this case, each preparation preferably contains, for example, 300 mg to 550 mg of daptomycin.
Containers such as vials and syringes may be made of either glass or plastic. Of these, glass or olefin-based plastics are preferred. As plastics, cycloolefin polymers, polyethylene and polypropylene are particularly preferred, and cycloolefin polymers and polypropylene are more preferred. When the above-mentioned daptomycin is contained, a container of 10 mL to 20 mL may be used, and a 10 mL container is preferably used.

Generally, it is preferable to store the vial, syringe, etc. in a package. It is also preferable to place an oxygen scavenger on the outside of the vial, syringe, etc.
The packaging material is preferably one with low water vapor and/or oxygen permeability. For example, it is more preferable that the oxygen permeability is 1 mL/ ^m2 -day-atm or less and/or the water vapor transmission rate is 1 g/m2-day or less. Examples of such packaging include resin bags made of various resins (including aluminum-deposited products) such as polypropylene, polyethylene, polystyrene, polyamide, polycarbonate, polyvinyl chloride, acrylic resin, polyester, and polyolefin, in a single layer structure or a laminated structure.

The daptomycin-containing lyophilized formulations of the present application can be supplied in the containers described above as sterile, pale yellow to light brown lyophilized cakes (lumps or powders) containing, for example, approximately 500 mg or 350 mg of daptomycin per formulation for intravenous use.
The daptomycin-containing lyophilized preparation of the present application can be redissolved in the above-mentioned aqueous medium as necessary and used as an injection solution, etc. In this case, the components and concentrations in these aqueous media can be those used for ordinary injections.
The pH of the solution containing daptomycin after reconstitution is preferably 5.5 to 7.5 from the viewpoint of drug stability.
The daptomycin-containing freeze-dried preparation of the present application can ensure stability over a long period of time even when stored at room temperature, and can be expected to dissolve quickly when reconstituted.

The freeze-dried formulation of the present application will be described in more detail below with reference to examples, although the present invention is not limited to the following examples.
Examples and Comparative Examples Drug solutions were prepared with the compositions shown in the following Table 1. As the stabilizers in the following Table 1, L-arginine, inositol, dextran 40, macrogol 4000, histidine, lactose, D-sorbitol, D-mannitol, xylitol and glycine were added in the amounts shown in Table 2.
First, daptomycin was dissolved in distilled water for injection (cooled to 10°C) to which sodium hydroxide (appropriate amount) had been added, the pH was adjusted to 4.7 by adding sodium hydroxide solution, and then the solution was diluted with distilled water for injection to obtain a daptomycin stock solution (daptomycin concentration: 110.25 mg/mL). Then, each additive was added and dissolved in the daptomycin stock solution in the amounts shown in Table 2, and further, a hydrochloric acid solution (appropriate amount) or a sodium hydroxide solution (appropriate amount) was added to adjust the pH to 6.8 to 7.1, and then the solution was diluted with distilled water for injection (daptomycin concentration: 91.875 mg/mL).
The resulting aqueous solution was sterilized by filtration, and the cooled aqueous solution was filled into a 10 mL glass vial (manufactured by Nipro) so that the vial contained 367.5 mg of daptomycin. The vial was then freeze-dried using a freeze-dryer, stoppered with a rubber stopper, and tightened with an aluminum cap to produce a freeze-dried preparation.


As a reference example, a lyophilized preparation was produced in the same manner as above except that the stabilizer was not included.
The obtained lyophilized preparation was stored at 60° C. for 21 days, after which the increase in the total amount of related substances was measured. The results are shown in Table 3. In Table 3, the increase in the amount of lyophilized preparation produced without using any additives was set to 1, and the values shown were calculated according to the following formula.
(1) Calculation of the increase: (each additive "total amount of related substances at 60°C for 21 days" - each additive "amount at the start") / (no additive "total amount of related substances at 60°C for 21 days" - no additive "amount at the start")

(2) Measurement method of total related substances The lyophilized preparation obtained in the Examples or Comparative Examples was redissolved in 7 mL of water. This solution was diluted 50-fold with a water/acetonitrile mixture (2:1), and 10 μL of the solution was used in the test to measure the change over time in the total related substances.
<Detection conditions>
Equipment name: High-performance liquid chromatograph Detector: Ultraviolet absorption photometer (wavelength: 221 nm)
Column: 250 x 4.6 mm I.D. Octyl silica gel
Column temperature: constant temperature around 30°C Mobile phase A: 0.34% ammonium phosphate buffer (pH 3.1)
Mobile phase B: acetonitrile Mobile phase delivery: The concentration gradient was controlled by changing the mixing ratio of mobile phase A and mobile phase B as follows:

Flow rate: 1mL/min.
Under the above detection conditions, the peak area of each sample was measured by automatic integration, and the percentage of the total related substances was obtained.

Of the additives shown in Table 2, L-arginine, inositol, dextran 40 and L-histidine were confirmed to significantly inhibit the increase in both total related substances and lactone hydrolysates compared to the lyophilized preparation produced without additives. Macrogol 4000 was confirmed to significantly inhibit the increase in total related substances compared to the lyophilized preparation produced without additives.

Test Example: Of the above stabilizers, L-arginine was found to have the most significant effect, and freeze-dried preparations were produced at the addition amounts of 350 mg, 400 mg, and 500 mg. For the specimen, L-arginine was dissolved in a hydrochloric acid solution at 10°C, after which daptomycin was dissolved and the pH was adjusted to 6.8 with a hydrochloric acid solution (sodium hydroxide was not used). Other than that, the freeze-dried preparations were produced in the same manner as in the above examples.
As a comparative example, a lyophilized preparation was produced by adding 524 mg of sucrose instead of L-arginine (since sucrose was described in Patent Document 1: JP-T-2013-511557). However, a sodium hydroxide solution was used instead of a hydrochloric acid solution, and the pH was adjusted to 7.0.
The freeze-dried preparation thus obtained was stored at 60° C. for 21 days, after which the increase in the total amount of related substances and the time required for redissolution were evaluated. The results are shown in Table 4, and the evaluation results of the increase in the total amount of related substances are shown in FIG.

As is clear from Table 4 and Figure 1, it was confirmed that when L-arginine was used as a stabilizer in a solution containing daptomycin, the generation of total related substances was suppressed and an increase in the total related substances over time could be prevented with an increase in the amount of L-arginine added. It was also confirmed that the addition of L-arginine allowed the daptomycin-containing lyophilized preparation to be redissolved relatively quickly.

The daptomycin-containing lyophilized formulation of the present application can be stabilized to the extent that it can be stored at room temperature.

Claims (3)

Daptomycin,
and a stabilizer which is L-arginine,
The content of L-arginine is 350 mg to 400 mg per 367.5 mg of daptomycin, and
The freeze-dried preparation contains no amino acids other than L-arginine. The lyophilized formulation of claim 1, which does not contain a calcium source. 3. The stable lyophilized preparation according to claim 1 or 2, wherein the pH of the preparation when dissolved with water for injection is 5.5 to 7.5.