Method for preparing solution for quantification of olamine

Abstract

To provide a method for preparing a solution for quantification of olamine serving as a counter ion in an oral solid preparation comprising eltrombopag olamine.SOLUTION: A method for preparing a solution for quantification of olamine according to one embodiment of the present invention comprises the steps of: (i) adding a solution having a pH of less than 2.5 to an oral solid preparation; (ii) subjecting the solution obtained in step (i) to ultrasonic treatment in a water bath or an organic solvent bath to obtain a suspension of the oral solid preparation and then, if the temperature of the suspension is below 10°C, heating the suspension to 10°C or higher; and (iii) filtering the suspension obtained in step (ii) to provide the filtrate as the solution for quantification of olamine.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for preparing a solution for quantifying olamine as a counter ion contained in an oral solid dosage form containing eltrombopag olamine. [Background technology]

[0002] In the presence of multivalent ions, eltrombopag olamine forms a complex with 2-aminoethanol (hereinafter referred to as "olamine"), converting it to eltrombopag without the counter ion olamine (hereinafter referred to as "eltrombopag free form"), resulting in a decrease in solubility. Because solubility has a significant effect on bioabsorption, understanding the content of eltrombopag olamine in a formulation is important for understanding the formulation's stability and dissolution characteristics, and ultimately for ensuring pharmaceutical quality.

[0003] For example, methods for quantifying eltrombopag olamine are disclosed in Non-Patent Documents 1 to 4. All of the methods for quantifying eltrombopag olamine disclosed in Non-Patent Documents 1 to 4 are methods for quantifying eltrombopag olamine as eltrombopag.

[0004] Furthermore, methods for quantifying olamine are known (for example, Non-Patent Documents 5 and 6). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Balaji Jayagopal et al., Journal of Pharmaceutical and Biomedical Analysis, (2021), vol. 203, Article 114231 [Non-patent document 2] Dharti Patel et al., World Journal of Pharmacy and Pharmaceutical Sciences, (2020), vol. 9, p1742-1751 [Non-patent document 3] Dr. Ajay I. Patel et al., Journal of the Turkish Chemical Society, Section A: Chemistry, (2020), vol. 7, p845-850 [Non-patent document 4] Madasu Raja Kumar et al., World Journal of Pharmacy and Pharmaceutical Sciences, (2018), vol. 7, p744-762 [Non-Patent Document 5] HPLC Separation of Ethanolamines on Amaze SCMixed-Mode Column [Retrieved March 8, 2024], Internet <URL:https: / / helixchrom.com / applications / hplc-separation-of-ethanolamines-on-amaze-sc-mixed-mode-column / > [Non-patent document 6] Shodex HPLC column product description page [searched March 8, 2024], Internet<URL:https: / / www.shodex.com / ja / dc / 08 / 02 / 08.html> Summary of the Invention [Problem to be solved by the invention]

[0006] Conventional methods for quantifying eltrombopag olamine cannot distinguish between eltrombopag olamine and free eltrombopag, which poses a problem in that the content of eltrombopag olamine in a pharmaceutical preparation cannot be accurately determined.

[0007] An object of one aspect of the present invention is to provide a technique for preparing a solution for quantifying olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine. [Means for solving the problem]

[0008] The present inventors focused on olamine as a counter ion and believed that if the content of olamine could be determined, the drawbacks of conventional methods for quantifying eltrombopag olamine could be overcome and the content of eltrombopag olamine in a formulation could be determined. As a result of extensive research aimed at solving the above-mentioned problems, the present inventors discovered for the first time that a sample solution for quantifying olamine as a counter ion in an oral solid formulation can be prepared by ultrasonically treating an oral solid formulation containing eltrombopag olamine in a solution of a specific pH, and then heating the resulting suspension at a specific temperature as necessary, thereby completing the present invention.

[0009] That is, in order to solve the above-mentioned problems, a method for preparing a solution for quantifying olamine according to one aspect of the present invention is a method for preparing a solution for quantifying olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine, the method comprising the steps of: (i) adding a solution with a pH of less than 2.5 to the oral solid formulation containing eltrombopag olamine; (ii) ultrasonically treating the solution obtained in step (i) in a water or organic solvent bath to obtain a suspension of the oral solid formulation, and then, if the suspension is at a temperature lower than 10°C, warming the suspension to 10°C or higher; and (iii) filtering the suspension of the oral solid formulation obtained in step (ii) and obtaining the filtrate as a solution for quantifying the olamine. Includes. [Effects of the Invention]

[0010] According to one aspect of the present invention, a technique for preparing a solution for quantifying olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine can be provided. DETAILED DESCRIPTION OF THE INVENTION

[0011] One embodiment of the present invention will be described in detail below. In this specification, unless otherwise specified, the expression "A to B" representing a range of numerical values ​​means "A or more and B or less."

[0012] [1. Preparation of solution for olamine determination] A method for preparing a solution for quantifying olamine according to one embodiment of the present invention (hereinafter, sometimes referred to as the "preparation method according to the present invention") is a method for preparing a solution for quantifying olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine, the method comprising the steps of: (i) adding a solution with a pH of less than 2.5 to the oral solid formulation containing eltrombopag olamine; (ii) ultrasonically treating the solution obtained in step (i) in a water or organic solvent bath to obtain a suspension of the oral solid formulation, and then, if the suspension is at a temperature lower than 10°C, warming the suspension to 10°C or higher; and (iii) filtering the suspension of the oral solid formulation obtained in step (ii) and obtaining the filtrate as a solution for quantifying the olamine. Includes.

[0013] According to a preparation method of one embodiment of the present invention, olamine derived from olamine as a counter ion can be extracted with high extraction efficiency from an oral solid formulation containing eltrombopag olamine. Furthermore, although the detailed reasons will be explained later, the olamine quantification solution obtained by the preparation method of one embodiment of the present invention contains only olamine that has dissociated from eltrombopag olamine after the start of preparation of the quantification solution. In other words, at the time of preparation of the quantification solution, free olamine (hereinafter sometimes referred to as "free olamine") that has already dissociated from eltrombopag olamine in the oral solid formulation is not present. Therefore, when olamine is quantified in the olamine quantification solution prepared by the preparation method of one embodiment of the present invention, the quantified olamine is olamine derived from olamine as a counter ion. In this specification, the term "olamine as a counter ion" refers to olamine present as a counter ion of eltrombopag. Furthermore, the term "olamine derived from olamine as a counter ion" refers to olamine that is dissociated from eltrombopag olamine after the start of preparation of the assay solution, and may be expressed as "counter ion olamine."

[0014] In this way, by using the olamine quantification solution prepared by the preparation method according to one embodiment of the present invention as a sample solution for quantifying olamine, it is possible to more accurately determine the content of olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine, and as a result, it is possible to more accurately determine the content of eltrombopag olamine.

[0015] The reason why free olamine is absent in the olamine quantification solution prepared by the preparation method according to one embodiment of the present invention is explained below. In oral solid formulations, olamine as a counter ion is thought to exist in an equilibrium state due to electrostatic interactions. On the other hand, free olamine in oral solid formulations is thought to be dissolved in ethanol and / or water, which are present in trace amounts in oral solid formulations. Furthermore, olamine is highly reactive because it possesses the properties of both an amine and an alcohol. Therefore, free olamine generated in oral solid formulations is thought to react rapidly and irreversibly with additives, impurities, etc. contained in the oral solid formulation.

[0016] Furthermore, the operation of extracting counter ion olamine from eltrombopag olamine, which is bound by electrostatic interaction (ionic bond) in the production method according to one embodiment of the present invention, does not involve the dissociation energy required to cleave the covalent bond, and therefore it is believed that it is possible to extract only counter ion olamine from eltrombopag olamine.

[0017] From the above, free olamine is not present in the oral solid formulation. Therefore, it can be said that free olamine is not present in the olamine quantification solution according to one embodiment of the present invention prepared using such an oral solid formulation. Note that although the case where the oral solid formulation is, for example, a tablet has been described, the same is considered to apply when the oral solid formulation is in a dosage form other than a tablet.

[0018] (eltrombopag olamine) Eltrombopag olamine has the chemical name 3'-{(2Z)-2-[1-(3,4-dimethylphenyl)-3-methyl-5-oxo-1,5-dihydro-4H-pyrazol-4-ylidene]hydrazino}-2'-hydroxybiphenyl-3-carboxylic acid bis(2-aminoethanol) and the molecular formula C 25 H 22 N4O4·2(C2H7NO), a compound with a molecular weight of 564.6.

[0019] In the presence of polyvalent ions, olamine forms a complex, resulting in free eltrombopag, which reduces the solubility of eltrombopag olamine. [ka]

[0020] Eltrombopag olamine can be produced by known methods, such as the production method described in WO 03 / 098992. Additionally, oral solid formulations containing eltrombopag olamine are commercially available, for example, as Revolade (registered trademark) tablets (manufactured by Novartis Pharma K.K.).

[0021] In one embodiment of the present invention, a method for preparing an olamine quantification solution is prepared using an oral solid formulation containing eltrombopag olamine. The dosage form of the oral solid formulation containing eltrombopag olamine is not particularly limited, and any dosage form of oral solid formulation is applicable. Examples of such dosage forms include tablets, granules, powders, and hard capsules. Tablets may be coated with a conventional coating agent.

[0022] The oral solid formulation containing eltrombopag olamine may contain ingredients such as additives that are commonly used in the pharmaceutical field, and may have a size that is commonly used in the pharmaceutical field.

[0023] Hereinafter, each step of a manufacturing method according to one embodiment of the present invention will be described in detail.

[0024] <Process (i)> Step (i) is a step of adding a solution having a pH of less than 2.5 to an oral solid formulation containing eltrombopag olamine. By adding a solution having a pH of less than 2.5 to an oral solid formulation containing eltrombopag olamine, counterion olamine can be extracted from the oral solid formulation containing eltrombopag olamine with high extraction efficiency. In the following description, for convenience, the "solution having a pH of less than 2.5" will be referred to as the "acid solution."

[0025] From the viewpoint of improving the extraction efficiency of eltrombopag olamine, the pH of the acid solution is preferably 2.0 or less, more preferably 1.8 or less, and there is no particular lower limit to the pH of the acid solution.

[0026] The type of acid solution is not particularly limited, and examples thereof include hydrochloric acid solution, sulfuric acid solution, nitric acid solution, and trifluoroacetic acid (TFA) solution.

[0027] The amount of acid solution added to the oral solid formulation containing eltrombopag olamine can be an amount that does not interfere with obtaining a suspension of the oral solid formulation during the ultrasonic treatment in the next step (ii). For example, the amount of acid solution added to the oral solid formulation containing eltrombopag olamine is preferably about 0.01 mL or more per 1 mg of oral solid formulation. On the other hand, if the amount of acid solution is too large, eltrombopag olamine may be too diluted to accurately quantify it, so the amount of acid solution is preferably about 0.8 mL or less per 1 mg of eltrombopag olamine.

[0028] <Process (ii)> Step (ii) is a step of ultrasonically treating the solution obtained in step (i) in a water or organic solvent bath to obtain a suspension of the oral solid formulation, and then, if the suspension is at a temperature lower than 10°C, warming the suspension to 10°C or higher.

[0029] The liquid poured into the ultrasonic generator for ultrasonic treatment can be water or an organic solvent that is liquid at room temperature and has a boiling point of about 75°C or higher, but water is preferred. When using an organic solvent, it is preferable to use ethanol or edible vegetable oil from the standpoint of safety.

[0030] The ultrasonic treatment conditions can be those under which a suspension of an oral solid preparation containing eltrombopag olamine is obtained from the oral solid preparation, and are preferably those under which the oral solid preparation containing eltrombopag olamine is completely disintegrated and a suspension in which the oral solid preparation is uniformly dispersed in a solution is obtained. Such conditions can be achieved, for example, by using a conventionally known ultrasonic treatment device and adjusting the output, pulse duration, treatment time, etc. of the ultrasonic treatment device depending on the dosage form, size, etc. of the oral solid preparation containing eltrombopag olamine.

[0031] If the suspension obtained by ultrasonic treatment is at a temperature below 10°C, it is heated to 10°C or higher. This improves the extraction efficiency of eltrombopag olamine. From the viewpoint of improving the extraction efficiency of eltrombopag olamine, it is more preferable to heat the suspension until it reaches 15°C or higher, more preferably until it reaches 20°C or higher, even more preferably until it reaches 30°C or higher, still more preferably until it reaches 40°C or higher, and even more preferably until it reaches 50°C or higher.

[0032] From the viewpoint of improving the extraction efficiency of eltrombopag olamine, it is preferable to maintain the temperature of the suspension at 10°C or higher for a specific period of time after the temperature reaches 10°C or higher. In this specification, such a period of time is referred to as the "temperature maintenance time." The temperature maintenance time is preferably 15 minutes or longer, and more preferably 20 minutes or longer.

[0033] The suspension in step (ii) can be heated by a conventionally known method, for example, by immersing a container containing the suspension in a water bath or oil bath set to a specific temperature. The method for heating the suspension is not particularly limited as long as it can reach 10°C or higher. For example, the suspension may be left at room temperature (20°C to 25°C) until the suspension reaches 10°C or higher.

[0034] <Step (iii)> Step (iii) is a step of filtering the suspension of the oral solid formulation obtained in step (ii) to obtain the filtrate as a solution for quantifying the olamine. The filtration method is not particularly limited, but filtration is preferably performed using a filter with a smaller pore size, such as a conventionally known membrane filter with a pore size of 0.45 μm or less.

[0035] The obtained filtrate can be used as a solution for quantifying olamine. The filtrate may be used as an undiluted solution for quantifying olamine, or may be diluted as necessary and used as a solution for quantifying olamine.

[0036] According to a preparation method of one aspect of the present invention, counter ion olamine can be extracted from an oral solid formulation containing eltrombopag olamine with high extraction efficiency. For example, according to a preparation method of one aspect of the present invention, the extraction efficiency of counter ion olamine calculated from the following formula (1) is 90% or more, preferably 97% or more. Extraction efficiency (%) = Content of counterion olamine in the filtrate obtained from the oral solid dosage form (measured value) / Content of eltrombopag olamine in the oral solid dosage form (theoretical value) × 100 (1)

[0037] Therefore, by quantifying the counter ion olamine using the olamine quantification solution prepared by the preparation method according to one embodiment of the present invention, it is possible to more accurately determine the content of eltrombopag olamine in an oral solid formulation.

[0038] <2. Olamine Quantitative Solution> The present invention also encompasses an olamine quantification solution prepared by the preparation method according to one embodiment of the present invention. By quantifying the counter ion olamine using the olamine quantification solution according to one embodiment of the present invention, it becomes possible to more accurately determine the content of eltrombopag olamine in an oral solid dosage form.

[0039] An example of a method for quantifying eltrombopag olamine using an olamine quantification solution according to one embodiment of the present invention will be described below.

[0040] First, one tablet of the oral solid formulation is used to prepare a solution for quantification of olamine by the preparation method according to one embodiment of the present invention. The prepared solution for quantification of olamine is used as a sample solution, and the peak area A of olamine is measured by liquid chromatography. T Similarly, for a separately prepared standard solution of olamine, the peak area A of olamine was determined using liquid chromatography. S The standard solution of olamine is prepared by the method described in the Examples below, and liquid chromatography is performed under the conditions described in the Examples below.

[0041] Olamine peak area A T and A S From this, the content (%) of olamine relative to the eltrombopag olamine (drug substance) contained in one tablet of the oral solid formulation can be calculated using the following formula (2): E=M S ×A T / A S / d / C×100 ···(2) E: The olamine content (%) of eltrombopag olamine (drug substance) contained in the oral solid dosage form used to prepare the olamine quantitative solution d: Dilution factor (where 1 is the volume of the container used to prepare the standard solution and z is the volume of the container used to prepare the olamine quantitation solution, then 25 / z. For example, if the volume of the container used to prepare the standard solution is 20 mL and the volume of the container used to prepare the olamine quantitation solution is 10 mL, then z is 1 / 2 and d is 50.) MS : Amount of olamine (mg) used to prepare the standard solution C: Theoretical content (mg) of eltrombopag olamine in one oral solid tablet (For example, eltrombopag olamine tablets 12.5 mg: 15.9, eltrombopag olamine tablets 25 mg: 31.8).

[0042] As described above, all of the olamine detected by liquid chromatography is counterion olamine, and therefore the olamine content (%) is the same as the eltrombopag olamine content (%).

[0043] In this way, by using the olamine quantification solution according to one embodiment of the present invention, the content of olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine can be determined more accurately, and as a result, the content of eltrombopag olamine can be determined more accurately. Note that the above-mentioned method is only one example of a method for quantifying eltrombopag olamine using the olamine quantification solution according to one embodiment of the present invention, and eltrombopag olamine contained in an oral solid formulation containing eltrombopag olamine may also be quantified by quantifying counter ion olamine using other methods.

[0044] After the start of preparation of the quantitative solution, a portion of the olamine dissociated from eltrombopag olamine may react with additives, impurities, and the like contained in the oral solid formulation. As can be seen from the extraction efficiency of counter ion olamine shown in the Examples below, the proportion of such "olamine reacted with additives" after dissociation is small. However, it is possible to measure "olamine reacted with additives" after dissociation by a test under different conditions from the counter ion olamine. By further performing such a test, the content of olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine can be more accurately determined.

[0045] The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope of the claims. Embodiments obtained by appropriately combining the technical means disclosed in the embodiments are also included in the technical scope of the present invention. [Example]

[0046] <Reagents and solutions> 2-aminoethanol (Fujifilm Wako Pure Chemical Industries, special grade) 0.1% trifluoroacetic acid solution / acetonitrile mixture (49:1) 0.1% trifluoroacetic acid solution 1 mL of trifluoroacetic acid was mixed with 1000 mL of water. Trifluoroacetic acid (Fujifilm Wako Pure Chemical Industries, special grade) Acetonitrile (Fujifilm Wako Pure Chemical Industries, high performance liquid chromatograph grade) Hydrophilic PTFE membrane filter, pore size 0.45 μm (ADVANTEC) - Silica gel column for liquid chromatography, bonded with alkyl groups having acidic groups, φ3.0 mm x 10 cm, 5 μm (HELIX AMAZE SC)

[0047] The details of the solutions added in step (i) were as follows: The pH was adjusted using sodium hydroxide. Since 0.1% TFA = 13 mM, the molar concentrations of the other acid solutions were also adjusted to 13 mM. Below are the measured pH values ​​of each acid solution. 13mM acetic acid: pH 3.37 13mM Hydrochloric Acid: pH 1.99 ·13mM sulfuric acid: pH1.79 0.1% TFA (=13 mM): pH 1.99

[0048] [1. Preparation of standard solutions] Approximately 172.5 mg of olamine was accurately weighed, and the mobile phase for liquid chromatography was added to make exactly 100 mL. 4 mL of this solution was accurately weighed, and the mobile phase was added to make exactly 20 mL, which was used as the standard solution.

[0049] 2. Preparation of sample solution

[0050] (Preparation Example 1) The oral solid formulation used to prepare the olamine quantitative solution was a plain tablet consisting of 31.8 mg of Eltrompovaguolamine, 14.9 mg of crystalline cellulose (PH102), 29.8 mg of D-mannitol (Mannit P), 3.2 mg of povidone K30 (Eifutact), 102.8 mg of crystalline cellulose (PH102), 100 mg of crystalline cellulose (KG802), 28 mg of sodium starch glycolate (Primogel), 24 mg of hydrogenated castor oil (Lubriwax-101), 12 mg of anhydrous calcium phosphate (Fujicalin), and 3.5 mg of magnesium stearate, film-coated with 14 mg of OPADRY WHITE.

[0051] One tablet of the above-mentioned oral solid formulation was taken and 10 mL of 0.1% TFA was added (step (i)). This solution was sonicated under the treatment conditions shown in Table 1 until complete disintegration of the tablet was confirmed visually, and the suspension after sonication was left at room temperature for 20 minutes (step (ii)). Thereafter, the mobile phase was further added to make the total volume exactly 20 mL. The resulting solution was filtered through a membrane filter with a pore size of 0.45 μm or less, and used as the sample solution of Preparation Example 1 (step (iii)).

[0052] (Preparation Examples 2 to 9, Comparative Preparation Examples 1 to 6) Sample solutions of Preparation Examples 2 to 9 and Comparative Preparation Examples 1 to 6 were prepared in the same manner as Preparation Example 1, except that steps (i) and (ii) were performed under the conditions shown in Table 1. For Preparation Examples 2 to 4 and 7 to 9, the suspensions after ultrasonic treatment were allowed to stand at the respective temperatures for 20 minutes. In the Preparation Examples and Comparative Preparation Examples other than Comparative Preparation Example 6, the temperature of the suspension after treatment in step (ii) was 10°C or higher.

[0053] 3. Quantitation of Olamine 10 μL of each sample solution and standard solution was tested by liquid chromatography under the following conditions, and the peak area A of olamine in each solution was T and AS asked for.

[0054] <Liquid chromatography test conditions> Detector: Differential refractometer (detector temperature: constant temperature around 40°C) Column: A stainless steel tube having an inner diameter of 3.0 mm and a length of 10 cm was packed with 5 μm silica gel bonded with an alkyl group having an acidic group for liquid chromatography. Column temperature: constant temperature around 35°C Mobile phase: 0.1% trifluoroacetic acid solution / acetonitrile mixture (49:1) Flow rate: 1.0 mL per minute

[0055] <System Compatibility> System performance: When operating under the above conditions for 10 μL of standard solution, the theoretical plate number and symmetry factor of the olamine peak were 1500 or more and 0.8 to 13, respectively. System reproducibility: When the test was repeated six times using 10 μL of the standard solution under the above conditions, the relative standard deviation of the peak area of ​​olamine was 1.0% or less.

[0056] <Calculation of olamine content> For each sample solution of Preparation Examples 1 to 9 and Comparative Preparation Examples 1 to 6, the peak area A of olamine T and A S The content (%) of olamine relative to the eltrombopag olamine (drug substance) contained in one tablet of the oral solid formulation was calculated using the above-mentioned formula (2). The results are shown in Table 1.

[0057] [Table 1]

[0058] The results shown in Table 1 indicate that the extraction efficiency of olamine differs depending on the pH. In addition, the extraction efficiency of olamine differs depending on whether or not heating was performed after ultrasonic treatment in an ice-water bath.

[0059] From the above results, it was found that a solution for olamine quantification could be prepared by adding a solution of less than pH 2.5 to an oral solid formulation containing eltrombopag olamine in step (i), subjecting the resulting solution to ultrasonic treatment in step (ii), and subjecting the resulting suspension to step (iii). Furthermore, it was found that the use of this solution for olamine quantification could more accurately determine the content of olamine as a counter ion in an oral solid formulation containing eltrombopag olamine, thereby enabling more accurate determination of the content of eltrombopag olamine. [Industrial Applicability]

[0060] The present invention can be used to determine the content of eltrombopag olamine in an eltrombopag olamine-containing oral solid formulation.

Claims

[Claim 1] 1. A method for preparing a solution for quantifying olamine as a counter ion contained in an oral solid formulation containing eltrombopag olamine, comprising: (i) adding a solution having a pH of less than 2.5 to the oral solid formulation containing eltrombopag olamine; (ii) ultrasonically treating the solution obtained in the step (i) in a water or organic solvent bath to obtain a suspension of the oral solid formulation, and then, if the suspension is at a temperature lower than 10°C, warming the suspension to 10°C or higher; and (iii) filtering the suspension of the oral solid formulation obtained in the step (ii) and obtaining the filtrate as a solution for quantifying the olamine. A method for preparing a solution for quantifying olamine, comprising:

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