JP4906546B2 - Revoholinate-containing aqueous solution formulation - Google Patents

Description

  The present invention relates to a stabilized aqueous solution preparation containing levofolinate or a salt thereof as an active ingredient, and a method for stabilizing a levofolinate-containing aqueous solution.

  Levophorinate (1-LV) has the chemical name (6S) -N- [4-[[(2-amino-5-formyl-1,4,5,6,7,8-hexahydro-4-oxo-6- Pteridinyl) methyl] amino] benzoyl] -L-glutamic acid, which is used as an antidote for the antifolate methotrexate and as an antitumor effect enhancer for fluorouracil.

  In the chemical synthesis process of levofolinate, the racemic body (leucovorin) containing equal amounts of d-form and 1-form is obtained by the configuration of the 6-position carbon atom, but only one has biological activity. It has been reported that levofolinate calcium, which is a calcium salt of a compound obtained by removing the non-active d-form from leucovorin, has been marketed (trade name: Isoborin Injection (Takeda Pharmaceutical)).

  However, since levofolinate calcium is usually extremely unstable in aqueous solutions, the only preparations that have been marketed so far are lyophilized preparations, and the aqueous preparations must be prepared at the time of administration. There has been a demand for the development of an aqueous solution preparation that can stably dissolve salts and can be stored for a long period of time.

  Moreover, although the effective concentration of levofolinate calcium is relatively high, a preparation containing a sufficient amount of active ingredient in one vial has not been put on the market. Therefore, when preparing an aqueous solution preparation with an effective concentration at the time of use, a plurality of vials are used. However, since this operation is extremely complicated, development of an aqueous solution preparation containing levofolinate at a high concentration is desired. It was.

As an attempt to improve the stability of hololinates or salts thereof in an aqueous solution, a mixture comprising leucovorin salts, tromethamine (buffering agent), and monothioglycerol (antioxidant) (Patent Document 1), hololinates Prefilled syringe preparation (Patent Document 2) filled with an aqueous solution containing as an active ingredient, 5-formyl- (6S) -tetrahydrofolic acid or a salt thereof as an active ingredient, and a pH adjuster and an antioxidant were blended Although an aqueous solution preparation for injection (Patent Document 3) has been reported, a sufficient effect has not been obtained for any of these.
Japanese Patent Laid-Open No. 3-90026 JP 2006-25953 A JP 2006-111614 A

  An object of the present invention is to provide a levofolinate-containing high concentration aqueous solution preparation in which levofolinate or a salt thereof is stably dissolved and can be stored for a long period of time, and a method for stabilizing levofolinate-containing high concentration aqueous solution.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have surprisingly found that levofolinate or a salt thereof has 4- (2-hydroxyethyl) -1-piperazine ethanesulfonic acid (hereinafter referred to as HEPES). Or a salt thereof, piperazine-1,4-bis (2-ethanesulfonic acid) (hereinafter referred to as PIPES) or a salt thereof, and one or more compounds selected from nicotinamide, an aqueous solution of levofolinate or a salt thereof It was found that the aqueous solution formulation containing levofolinate that can be stored for a long period of time was obtained, and the present invention was completed.

That is, the present invention provides the following components (A) and (B);
(A) one or more compounds selected from (B) (i) HEPES or a salt thereof, (ii) PIPES or a salt thereof, and (iii) nicotinic acid amide. An aqueous solution preparation containing levofolinate is provided.

  The present invention also provides a stable levofolinate-containing aqueous solution characterized by adding one or more compounds selected from HEPES or a salt thereof, PIPES or a salt thereof, and nicotinamide to an aqueous solution containing levofolinate or a salt thereof. It provides a method.

  In the aqueous solution preparation of the present invention, levofolinate or a salt thereof can be dissolved at a high concentration and stably and can be stored for a long period of time, and even when used clinically, it is not necessary to dissolve the solid preparation over time, It can be administered to the patient immediately, or can be administered simply by diluting to an appropriate concentration with a diluent. Moreover, the fall of the pH value of the aqueous solution at the time of storage is suppressed.

  The component (A) levofolinate or a salt thereof used in the aqueous solution preparation of the present invention is an active ingredient of the aqueous solution preparation, and is preferably used in the form of a salt in the present invention.

  As the salt of levofolinate used in the present invention, alkali metal salts such as sodium salt and potassium salt, or alkaline earth metal salts such as calcium salt and magnesium salt can be preferably used, and calcium salt is particularly preferable. These salts include both anhydrous salts that are anhydrides and hydrated salts that are hydrates.

  The salt of the component levofolinate used in the present invention may be formed in an aqueous preparation by adding levofolinate and an alkaline agent. In this case, examples of the alkali agent include sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, sodium carbonate, sodium hydrogen carbonate and the like, among which calcium hydroxide is preferable. In the aqueous preparation, a salt of levofolinate may be formed by salt exchange with other compounds.

  Component (B) used in the aqueous solution preparation of the present invention is at least one selected from HEPES or a salt thereof, PIPES or a salt thereof, and nicotinamide, and among them, HEPES or a salt thereof and nicotinamide are preferable.

  As a salt of HEPES and PIPES, an alkali metal salt such as a sodium salt can be suitably used, and a sodium salt is preferable, and HEPES-1 sodium salt (hereinafter referred to as HEPES-Na), PIPES-1 sodium salt (hereinafter referred to as “HEPES-1 sodium salt”) PIPES-Na), PIPES-1.5 sodium salt (hereinafter referred to as PIPES-1.5Na) and the like are preferable.

  HEPES or a salt thereof (HEPES conversion) is contained in an amount of 0.1 mg to a saturated amount, and further 0.1 to 670 mg of levofolinate or a salt thereof with respect to 100 mg of levofolinate or a salt thereof (revophorinate conversion) in the aqueous solution preparation of the present invention. It is preferable in terms of improving the stability of the salt in an aqueous solution, and considering the safety as a pharmaceutical product, 0.1 to 50 mg, more preferably 0.1 to 6 mg, and particularly preferably 0.1 to 1.5 mg is preferable.

  In the aqueous solution of levofolinate or a salt thereof, PIPES or a salt thereof (PIPES conversion) contains 1 mg to a saturated amount, and further 1 mg to 7 g, with respect to 100 mg of levofolinate or a salt thereof (revophorinate conversion) in the aqueous solution preparation of the present invention It is preferable in terms of stability improvement in the case of 1 to 500 mg, more preferably 1 to 40 mg in view of safety as a pharmaceutical product.

  The stability of levofolinate or a salt thereof in an aqueous solution is 10 mg to a saturated amount and further 10 mg to 20 g of nicotinamide relative to 100 mg of levofolinate or a salt thereof (revophorinate equivalent) in the aqueous solution preparation of the present invention. In view of safety as a pharmaceutical, it is preferable to contain 10 mg to 1 g, further 10 to 250 mg, particularly 10 to 62.5 mg.

The aqueous preparation of the present invention preferably further contains one or more compounds selected from the group of triethanolamine, meglumine and trometamol for the purpose of improving the stability of levofolinate or a salt thereof in an aqueous solution. .
More preferably, HEPES and at least one compound selected from the group of triethanolamine, nicotinamide and trometamol are contained, and particularly preferably HEPES, triethanolamine and nicotinamide are contained. .

  For 100 mg of levofolinate or a salt thereof (revophorinate conversion) in the aqueous solution preparation of the present invention, triethanolamine is contained in an amount of 1.5 mg to a saturated amount, and further 1.5 mg to 20 g in an aqueous solution of levofolinate or a salt thereof. From the viewpoint of improving the stability, it is preferable to contain 1.5 to 500 mg, more preferably 1.5 to 30 mg, and particularly preferably 1.5 to 7.5 mg in view of safety as a pharmaceutical product.

  Meglumine is contained in an amount of 100 mg to a saturated amount, and further 100 mg to 20 g, with respect to 100 mg of levofolinate or a salt thereof (revophorinate conversion) in the aqueous solution preparation of the present invention, which improves the stability of levofolinate or a salt thereof in an aqueous solution In view of safety as a pharmaceutical, it is preferable to contain 100 mg to 15.4 g, and more preferably 100 mg to 3.85 g.

  The amount of trometamol in the aqueous solution preparation of the present invention containing 100 mg (converted to levofolinate) of trometamol in the range of 3.5 mg to a saturated amount, and further 3.5 mg to 20 g is stable in the aqueous solution of levofolinate or a salt thereof. From the viewpoint of improving the safety, it is preferable to contain 3.5 mg to 1 g, further 3.5 to 72 mg, particularly 3.5 to 18 mg in view of safety as a pharmaceutical product.

  The pH of the aqueous solution preparation of the present invention is preferably 9 to 11, more preferably 9 to 10, more preferably 9.2 to 10, particularly 9.5 from the viewpoint of improving the stability of levofolinate or a salt thereof in an aqueous solution. Here, the pH of the water-soluble preparation means the pH at the preparation of the preparation, and is a value at room temperature (25 ° C.). The pH adjustment is preferably performed using an acid such as acetic acid, hydrochloric acid or sulfuric acid, or an alkali such as sodium hydroxide, sodium carbonate or sodium bicarbonate.

  The dissolved oxygen concentration in the aqueous preparation of the present invention is preferably 1 mg / L or less, and particularly preferably 0.5 mg / L or less from the viewpoint of improving the stability of levofolinate or a salt thereof in an aqueous solution. In order to remove dissolved oxygen, it is preferable to replace the oxygen in the container space and the aqueous solution with an inert gas such as nitrogen. For example, a method in which an aqueous solution preparation is bubbled with nitrogen and filling into a container under a nitrogen atmosphere is suitably used. Examples of the inert gas include nitrogen or a rare gas such as helium, argon, neon, and nitrogen is particularly preferable.

  The aqueous solution preparation of the present invention is useful as an active folic acid preparation because levofolinate or a salt thereof as an active ingredient has an excellent fluorouracil enhancing effect. Examples of target malignant tumors include gastric cancer and colorectal cancer.

  The dosage form of the aqueous solution preparation of the present invention is preferably an injection preparation, particularly an intravenous preparation. In addition to the above components, injectable distilled water, glucose, mannose, lactose, saccharides typified by inositol, inorganic salts typified by sodium chloride, and other stabilizers and excipients that are usually used for injections. Components such as an agent and a buffer may be used. In the preparation for injection, levofolinate or a salt thereof is preferably contained in an amount of 2-50 mg / mL, more preferably 5-50 mg / mL in terms of levofolinate. Moreover, since the aqueous solution preparation of the present invention is stable even when levofolinate or a salt thereof is blended at a high concentration, levofolinate or a salt thereof can be 5 mg / mL or more in terms of levofolinate.

  The aqueous solution preparation of the present invention further contains a good buffer other than those described above, for example, 2-morpholinoethanesulfonic acid, monohydrate (hereinafter referred to as MES), bis (2-hydroxyethyl) iminotris (hydroxymethyl) methane (hereinafter referred to as “methyl methane”). , Bis-Tris), N- (2-acetamido) iminodiacetic acid (hereinafter referred to as ADA), N- (2-acetamido) -2-aminoethanesulfonic acid (hereinafter referred to as ACES), 2-hydroxy-3 -Morpholinopropane sulfonic acid (hereinafter referred to as MOPSO), N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid (hereinafter referred to as BES), 3-morpholinopropane sulfonic acid (hereinafter referred to as MOPS), N-tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid (hereinafter referred to as TES) ), 3- [N, N-bis (2-hydroxyethyl) amino] -2-hydroxypropanesulfonic acid (hereinafter referred to as DIPSO), 2-hydroxy-N-tris (hydroxymethyl) methyl-3-aminopropanesulfone Acid (hereinafter referred to as TAPSO), piperazine-1,4-bis (2-hydroxy-3-propanesulfonic acid), dihydrate (hereinafter referred to as POPSO), 2-hydroxy-3- [4- (2- Hydroxyethyl) -1-piperazinyl] propanesulfonic acid, monohydrate (hereinafter referred to as HEPPSO), 3- [4- (2-hydroxyethyl) -1-piperazinyl] propanesulfonic acid (hereinafter referred to as EPPS), N -[Tris (hydroxymethyl) methyl] glycine (hereinafter referred to as Tricine), N, N-bis (2-hydroxy Til) glycine (hereinafter referred to as Bicine), N-tris (hydroxymethyl) methyl-3-aminopropanesulfonic acid (hereinafter referred to as TAPS), N-cyclohexyl-2-aminoethanesulfonic acid (hereinafter referred to as CHES), N -Cyclohexyl-2-hydroxy-3-aminopropanesulfonic acid (hereinafter referred to as CAPSO), N-cyclohexyl-3-aminopropanesulfonic acid (hereinafter referred to as CAPS), and the like. Among these, MES, Bis-Tris, ADA, ACES, MOPSO, BES, MOPS, TES, DIPSO, TAPSO, POPSO, HEPPSO, EPPS, and the like are preferable. With respect to 100 mg of levofolinate or a salt thereof (revophorinate conversion) in the aqueous solution preparation of the present invention, these good buffers contain 0.1 mg to a saturated amount, and further 0.1 mg to 10 g. An aqueous solution of levofolinate or a salt thereof It is preferable from the viewpoint of improving the stability in the medium, and 0.1 mg to 1 g, more preferably 0.1 to 100 mg is particularly preferable in consideration of safety as a pharmaceutical product.

  If necessary, the aqueous solution preparation of the present invention may further contain isotonic agents (glucose, sodium chloride, potassium chloride, glycerin, mannitol, etc.), soothing agents (glucose, inositol, benzyl alcohol, mepivacaine hydrochloride, xylocaine hydrochloride, hydrochloric acid) Procaine, carbocaine hydrochloride, glycerin, propylene glycol, lidocaine hydrochloride, etc.) and preservatives (paraoxybenzoic acid esters such as methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, thimerosal, chlorobutanol, benzyl alcohol, etc.) be able to. These addition amounts can be easily set by those skilled in the art according to known techniques.

  In order to protect the aqueous preparation of the present invention from light, a light-shielding container such as amber glass may be used. Furthermore, if necessary, a light stabilizer may be added, and pharmaceutically acceptable light stabilizers can be widely used. For example, sugars such as glucose, sucrose, fructose, maltose, xylitol, sorbitol And sugar alcohols such as mannitol and polyhydric alcohols such as glycerin and propylene glycol. In addition, benzotriazole derivatives, benzophenone derivatives, salicylic acid derivatives, paraaminobenzoic acid derivatives, cinnamic acid derivatives, urocanic acid derivatives, riboflavin, folic acid, and the like are also suitable light stabilizers. One or more of these light stabilizers can be added to the aqueous solution preparation of the present invention, and the amount added is 0.01 to 1 w / v%, preferably 0.05 to 0.1 w / v%. It is.

  The aqueous solution preparation of the present invention may further contain an antioxidant as necessary. As the antioxidant, ascorbic acid or a salt thereof, sodium pyrosulfite, cysteine hydrochloride, sodium bisulfite, thioglycerol, thioglycolic acid or a salt thereof, sodium sulfite, or the like is used. The salt to be exemplified is exemplified. One or more of these antioxidants can be blended in the aqueous solution preparation of the present invention, and the amount added can be easily set by those skilled in the art according to known techniques.

  The aqueous solution preparation of the present invention may contain other drugs (anti-neoplastic agents such as irinotecan hydrochloride (CPT-11), oxaliplatin, 5-fluorouracil, uracil, mitomycin, etc.) as necessary, Other drugs may be packaged separately to form the aqueous solution preparation and kit product of the present invention.

  In the present invention, an aqueous solution preparation for injection is not limited to an injection in a final form, but an injection solution precursor preparation (liquid injection) that can be prepared into a final injection solution by using a solution (diluent) at the time of use. , Concentrated injection, etc.).

  The aqueous solution preparation of the present invention can be produced by a method known per se. For example, the pH of an aqueous solution containing levofolinate calcium, HEPES, triethanolamine, and nicotinamide is adjusted to a predetermined value, and then diluted to a predetermined levofolinate calcium concentration. Thereafter, it can be produced by sterilizing and filtering under aseptic conditions, filling a container such as an ampoule, vial, syringe (prefilled syringe) and sealing.

  By the method of the present invention, the aqueous solution containing levofolinate is stabilized by adding one or more compounds selected from HEPES or a salt thereof, PIPES or a salt thereof, and nicotinamide to an aqueous solution containing levofolinate or a salt thereof. be able to.

  Hereinafter, the content of the present invention will be described in more detail with reference to examples, but the present invention is not limited by these.

Example 1 Stabilizing effect of various additives (1) Sample preparation and test method A 5 mg or 20 mg (as levofolinate) / mL aqueous solution of levofolinate calcium is prepared by sonication and stirring. HEPES, PIPES-1.5Na and nicotinamide were added to this solution, adjusted to pH 9 by adding sodium hydroxide solution, stored at 60 ° C, sampled every week, diluted with 50% methanol and analyzed by HPLC. did.

(2) HPLC analysis conditions Column: A stainless tube having an inner diameter of 4.6 mm and a length of 15 cm is filled with 5 μm of octadecylsilylated silica gel for liquid chromatography (L-column ODS 5 μm, 4.6 × 150 mm).
Column temperature: constant temperature around 45 ° C. mobile phase: 0.008 mol / L sodium dihydrogen phosphate solution / methanol / tetrabutylammonium hydroxide reagent mixture (385: 110: 4) to which phosphoric acid is added to adjust pH to 7. Flow rate adjusted to 5: Adjust so that the retention time of revoholinate is about 10 minutes (about 1.0 mL / min)
Detector: UV absorptiometer (measurement wavelength: 254 nm)

(3) Results As shown in Table 1, an aqueous solution of levofolinate calcium was stored at 60 ° C. and a stability test was performed. In the case of water for injection alone, a precipitate was observed after 2 weeks, but when various additives were added to this, the stability was remarkably improved, especially when HEPES and PIPES-1.5Na were added, after 4 weeks. It was also confirmed that there was an extremely excellent stabilization effect.

Example 2 Stability evaluation test (examination of pH)
(1) Sample preparation and test method Add levofolinate calcium to an aqueous solution of each additive shown in Table 2, and stir in warm water at 50 ° C to dissolve well. To this is added sodium hydroxide solution to adjust the pH to 8.0, 9.0, 10.0 and 11.0. This solution was stored at 60 ° C. for 2 weeks and sampled every week. The sampled solution was diluted with 50% methanol and analyzed by HPLC (HPLC conditions are according to Example 1).

(2) Results As shown in Table 2, the aqueous solution of levofolinate calcium was stored at 60 ° C. and a stability test was performed. When HEPES was added alone and when nicotinamide was further added in combination, the stabilizing effect of the latter formulation tended to be improved in all pH ranges, but both showed pH 9.0. In the case of ˜10.0, an extremely excellent stabilizing effect was shown. In addition, it was confirmed that a sufficient stabilizing effect was exhibited even with a high concentration levofolinate-containing aqueous solution of 20 mg / mL.

Example 3 Stability evaluation test (examination of prescription)
(1) Sample preparation and test method Add levofolinate calcium to the aqueous solution of each additive shown in Tables 3 to 5, and stir in warm water at 50 ° C to dissolve well. Sodium hydroxide solution is added to this to adjust the pH to 9.5. This solution was stored at 60 ° C. for 1 month and sampled every week. The sampled solution was diluted with 50% methanol and analyzed by HPLC (HPLC conditions are according to Example 1).

(2) Results As shown in Table 3, an aqueous solution of levofolinate calcium (5 mg / mL as levofolinate) was stored at 60 ° C. and a stability test was performed. In the case of water for injection alone, the residual rate of levofolinate was about 70% or less after 2 weeks, and a precipitate was observed after 3 weeks. When HEPES was added thereto, no precipitation was observed after 4 weeks, and the residual rate of levofolinate was 90% or more. By adding triethanolamine in addition to HEPES, the residual rate at 4 weeks increased to 92% or more, and by adding nicotinamide, a higher residual rate (about 94%) was obtained.

  As shown in Tables 4 and 5, aqueous solutions of levofolinate calcium (10 mg / mL and 20 mg / mL as levofolinate) were similarly stored at 60 ° C. and subjected to a stability test. Although there was a tendency for the residual rate to decrease as the concentration increased, compared with the case of no additive (only water for injection), a very excellent stabilizing effect was recognized by adding HEPES alone, and another 1 By adding a combination of more than seed additives, the stabilizing effect was further improved.

Example 4
The injections of Examples 1 to 6 below were obtained by the following production method.
122 mg of levofolinate calcium is added to 18 mL of an aqueous HEPES solution adjusted to a concentration of 0.067 mg / mL, and dissolved well in 50 ° C. warm water. Additives were added to this solution and dissolved by stirring. Each additive was added to this solution and dissolved, then the pH was adjusted to 9.5 with a sodium hydroxide solution, and the HEPES aqueous solution was added to make 20 mL.

Example 1
Revoholinate calcium 122mg
HEPES 1.34mg
Sodium hydroxide appropriate amount Water for injection Total 20mL
pH 9.5
Dissolved oxygen concentration 0.5mg / L

Example 2
Revoholinate calcium 122mg
HEPES 1.34mg
Triethanolamine 6.83mg
Sodium hydroxide appropriate amount Water for injection Total 20mL
pH 9.5
Dissolved oxygen concentration 0.5mg / L

Example 3
Revoholinate calcium 122mg
HEPES 1.34mg
Triethanolamine 6.83mg
Nicotinamide 57.3mg
Sodium hydroxide appropriate amount Water for injection Total 20mL
pH 9.5
Dissolved oxygen concentration 0.5mg / L

Example 4
Revoholinate calcium 122mg
HEPES 1.34mg
Triethanolamine 6.83mg
Trometamol 15.8mg
Sodium hydroxide appropriate amount Water for injection Total 20mL
pH 9.5
Dissolved oxygen concentration 0.5mg / L

Example 5
Revoholinate calcium 122mg
HEPES 1.34mg
Triethanolamine 6.83mg
Nicotinamide 57.3mg
Trometamol 15.8mg
Sodium hydroxide appropriate amount Water for injection Total 20mL
pH 9.5
Dissolved oxygen concentration 0.5mg / L

Example 6
Revoholinate calcium 122mg
PIPES-1.5Na 20mg
Meglumine 1000mg
Trometamol 15.8mg
Sodium hydroxide appropriate amount Water for injection Total 20mL
pH 9.5
Dissolved oxygen concentration 0.5mg / L

  The levofolinate-containing aqueous solution preparations (injections) of Examples 1 to 6 were all slightly yellowish clear aqueous solutions, and no crystal precipitation and decomposition of levofolinate were observed.

Claims (10)

The following components (A) and (B):
(A) Levophorinate or a salt thereof (B) (i) 4- (2-hydroxyethyl) -1-piperazine ethanesulfonic acid or a salt thereof (ii) Piperazine-1,4-bis (2-ethanesulfonic acid) or a salt thereof A levofolinate-containing aqueous solution preparation comprising a salt, and (iii) one or more compounds selected from the group of nicotinamide.   Furthermore, pH is 9-11, The levofolinate containing aqueous solution formulation of Claim 1 characterized by the above-mentioned.   The levofolinate-containing aqueous preparation according to claim 1 or 2, further comprising one or more compounds selected from the group consisting of triethanolamine, meglumine, and trometamol.   The levofolinate-containing aqueous solution preparation according to any one of claims 1 to 3, wherein the salt of levofolinate is an alkali metal salt or an alkaline earth metal salt.   The levofolinate-containing aqueous solution preparation according to any one of claims 1 to 4, wherein the salt of levofolinate is a calcium salt.   It is an aqueous solution preparation for injection, The levofolinate-containing aqueous solution preparation according to any one of claims 1 to 5.   The levofolinate-containing aqueous solution preparation according to any one of claims 1 to 6, wherein the concentration of levofolinate or a salt thereof is 5 mg / mL or more in terms of levofolinate.   8. The levofolinate-containing aqueous solution preparation according to any one of claims 1 to 7, wherein the dissolved oxygen concentration in the aqueous solution preparation is 1 mg / L or less.   4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid or a salt thereof, piperazine-1,4-bis (2-ethanesulfonic acid) or a salt thereof, and nicotinamide in an aqueous solution containing levofolinate or a salt thereof One or more types of compounds chosen from these are added, The stabilization method of the aqueous solution containing a levofolinate characterized by the above-mentioned.   Furthermore, pH is adjusted to 9-11, The stabilization method of the aqueous solution containing a levofolinate of Claim 9 characterized by the above-mentioned.