JP3501471B2 - Stabilizing composition and method of calcitonin - Google Patents

Description

Detailed Description of the Invention

[0001]

The present invention relates to calcitonins (Ca
The present invention relates to a stabilized composition which is a lyophilized product containing lcitonins) as an active ingredient and a method for stabilizing calcitonins.

[0002]

2. Description of the Related Art Calcitonins are polypeptide hormones involved in blood calcium concentration, and are drugs used for improving pain in hypercalcemia, Paget's disease of bone or osteoporosis, and natural calcitonin and its analogs. It has been known. These calcitonins,
A method of incorporating a saccharide such as mannitol as an excipient is generally used in preparing a freeze-dried preparation that is dissolved before use (JP-A-2-111).

Further, when calcitonin is formulated as an injectable solution, it contains sodium chloride as an isotonicity agent, an acetate buffer as a buffer, a citrate buffer and a lactate buffer, and 5-200 units of calcitonin as a main ingredient. Known injection preparations are freeze-dried preparations, in which excipients such as lactose, mannitol, dextran, and glycine are dissolved in these solutions in an amount of 2 to 3% and freeze-dried. 3-75532).

[0004]

Mannitol is the most widely used as an excipient for a freeze-dried preparation for dissolution of peptides, proteins and the like, but mannitol is a lyophilized preparation for dissolution-in-use of calcitonin. Even if it is used as an excipient, satisfactory results cannot be obtained regarding its stabilizing effect. Further, for example, generally, for the purpose of isotonicity, 0.
Freeze-dried injections of calcitonins with a sodium chloride concentration of about 7-0.9% cause the shrinkage of the cake due to too much sodium chloride, making it difficult to obtain the desired freeze-dried preparation. Then, as an excipient,
Even if 3% of sugars and amino acids were added, the same drawback could not be solved, and a freeze-dried composition containing calcitonin as an active ingredient and having good stability in use and a method for stabilizing calcitonin. Was wanted.

[0005]

[Means for Solving the Problems] To solve these problems, in order to obtain a safe and stable pharmaceutical formulation containing calcitonin as an active ingredient, as a result of various studies, surprisingly, various saccharides were 1/100 to 1/4 with respect to 1 part by weight of sugar
It was found that the stability of the freeze-dried preparation is remarkably improved by blending parts by weight of sodium chloride and freeze-drying as compared with the case of using the saccharide alone. As a result, by using saccharides and sodium chloride in combination as a stabilizer of a lyophilized formulation of calcitonin, it is safe and excellent in heat and storage stability, and also a lyophilized formulation of calcitonin which has no problem in appearance, Alternatively, it became possible to prepare a lyophilized composition.

That is, the present invention is a lyophilized product containing calcitonin as an active ingredient, containing a saccharide and sodium chloride, and having 1/100 to 1/4 parts by weight of sodium chloride to 1 part by weight of the saccharide. After dissolving a lyophilized composition characterized by being present, and calcitonin as an active ingredient, saccharides and sodium chloride in 1/100 to 1/4 parts by weight of sodium chloride per 1 part by weight of saccharides in an aqueous medium. It is a method for stabilizing calcitonins characterized by freeze-drying.

[0007] First, the calcitonin which is the active ingredient of the present invention is a polypeptide having at least a serum calcium lowering action, and a natural calcitonin or an analogue thereof is known. Examples of natural calcitonin include eel calcitonin, salmon calcitonin, porcine calcitonin, human calcitonin, chicken calcitonin and the like, which are secreted from endocrine cells and serve as calcium-regulating hormone. As the analog, an analog in which the S—S bond of natural calcitonin is replaced with a CH ₂ —CH ₂ bond by aminosuberic acid, for example, elcatonin ([Asu ^1.7 ] eel calcitonin), [Asu ^1.7 ] chicken calcitonin, [ Asu
^1.7 ] salmon calcitonin and [Asu ^1.7 ] human calcitonin are known.

[0008] Calcitonins are generally effective in a very small amount. For example, in elcatonin, a single dose of elcatonin having an activity of 6000 units / mg may be intravenously administered at about 10 to 40 units. In these calcitonins, an amount of 1 unit to 5000 units may be appropriately used in general.

Examples of the saccharides used in the present invention include monosaccharides, water-soluble polysaccharides of disaccharides or more, such as mannitol,
Examples thereof include water-soluble sugars such as glucose, sorbitol, inositol, xylitol, galactose, fructose, sucrose, maltose, lactose, trehalose, dextran and cyclodextrin. These saccharides may be used alone or in combination of two or more, and are not limited to these saccharides.

The amount of saccharide added is preferably 2 or more, more preferably 20 to 2 with respect to 1 weight of calcitonin.
0000 weight may be added. However, the object of the present invention is a stabilized composition of calcitonins, and since this composition is not necessarily limited to those used as a preparation for intravenous administration, it can be used as a solution in a container such as a vial or ampoule. The adjusted product is dispensed and then freeze-dried, and the amount of calcitonin as a main drug used at this time is usually 1 unit to 5000 units. On the other hand, the saccharide used in the present invention is, of course, used as a stabilizer during freezing,
A fixed amount may be added as an appropriate extender.

For this reason, it would be more realistic to define the amount of saccharide added in terms of the amount per ampoule or vial, rather than the relative ratio to the main drug, calcitonin. In this case, the amount added is about 0.5 to 100 mg, more preferably 1 to 20 mg. It is clear that the amount of sugar added in any case is not limiting in any way.

In the present invention, the amount of sodium chloride added is defined by the relative ratio with the saccharide. Calcitonins are more stable as the amount of sodium chloride added increases with respect to the saccharide, but when the weight ratio to the saccharide is 25% or more, shrinkage is observed in the cake after freeze-drying and the stability tends to decrease. Therefore, it is usually 1/100 to 1/4 parts by weight, preferably 1/100 parts by weight with respect to 1 part by weight of saccharides.
20 to 1/5 parts by weight may be used. Examples of the aqueous medium include distilled water for injection and a buffer solution. Further, the above-mentioned aqueous solvent may contain a water-soluble organic solvent, for example, a small amount of ethanol etc., as long as it does not exhibit toxicity.

To produce the freeze-dried stabilized composition for dissolution according to the present invention, for example, calcitonins, sugars and sodium chloride having the above-mentioned composition may be added, if necessary, to a well-known pH adjusting agent and a stabilizing agent. The agent, the bulking agent, the preservative and the like may be mixed, dissolved in the above aqueous medium, sterile filtered, and lyophilized according to a conventional method. For this freeze-drying, known freeze-drying methods such as tray freeze-drying, spray freeze-drying and vial freeze-drying can be adopted under the conditions usually used.

[0014]

The present invention will be described in detail below with reference to examples and comparative examples, but the present invention is not limited thereto.

[0015]

[Example 1] Elcatonin (manufactured by Asahi Kasei Corporation) 1.5 m
g (6000 units / mg), sucrose 500 mg and sodium chloride 50 mg, and sterile distilled water 50 m
1 was added and dissolved. After aseptic filtration, 0.5 ml each was dispensed into a glass vial, freeze-dried, replaced with nitrogen, and stoppered and wrapped to obtain a dry formulation for dissolution before use (invention product 1).

[0016]

Examples 2 to 6 In Example 1, the sugars shown in Table 1 were used in place of sucrose, and sodium chloride was treated in the same manner with the amounts shown in Table 1 to obtain each freeze-dried preparation (the present invention). Products 2 to 6) were obtained.

[0017]

[Table 1]

[0018]

[Example 7] Salmon calcitonin (manufactured by Sigma) 0.2 m
g, sucrose 100 mg and sodium chloride 10
10 mg of sterile distilled water was added and dissolved. Next, 0.2 ml each was dispensed into a glass vial, freeze-dried, and nitrogen substitution was performed, followed by stoppering and winding to obtain a soluble preparation for use. The obtained preparation had a good residual rate of calcitonin after 3 months at 40 ° C.

[0019]

Example 8 Eel calcitonin (manufactured by Sigma) 0.1
5 mg, maltose 180 mg and sodium chloride 9
mg was taken, and 9 ml of sterile distilled water was added and dissolved. Next, 0.1 ml each was dispensed into a glass ampoule, freeze-dried and then sealed to obtain a standard product for content measurement. The resulting formulation is 40
The calcitonin residual rate after 3 months at 0 ° C was good.

[0020]

[Example 9] Human calcitonin (manufactured by Sigma) 0.3 m
g, trehalose 100 mg and sodium chloride 10
10 mg of sterile distilled water was added and dissolved. Then, 0.5 ml each was dispensed into a glass vial, and after freeze-drying, it was stoppered and wound to obtain a dry formulation for dissolution before use. The obtained preparation had a good residual rate of calcitonin after 3 months at 40 ° C.

[0021]

EFFECT OF THE INVENTION Stability test of calcitonins

Preparation of Control Formulation

(1) Elcatonin (made by Asahi Kasei Corporation)
1.5 mg and 50 mg of sodium chloride were taken, and 50 ml of sterile distilled water was added and dissolved. After aseptic filtration, 0.5 ml each was dispensed into a glass vial, freeze-dried, replaced with nitrogen, capped, and tightened to obtain a dry formulation for dissolution before use (control product 1).

(2) Elcatonin (manufactured by Asahi Kasei Corporation)
1.5 mg and sucrose 500 mg were taken, and sterile distilled water 50 ml was added and dissolved. After aseptic filtration, 0.5 ml each was dispensed into a glass vial, freeze-dried, replaced with nitrogen, stoppered, and tightened to obtain a dry formulation for dissolution before use (control product 2).

(3) Elcatonin (made by Asahi Kasei Corporation)
1.5 mg and 500 mg of mannitol were taken, and 50 ml of sterile distilled water was added and dissolved. After aseptic filtration, 0.5 ml each was dispensed into a glass vial, freeze-dried to replace with nitrogen, and then capped and wound to obtain a dry formulation for dissolution before use (control product 3).

(4) Elcatonin (made by Asahi Kasei Corporation)
1.5 mg and lactose 500 mg were taken, and sterile distilled water 50 ml was added and dissolved. After aseptic filtration, 0.5 ml each was dispensed into a glass vial, freeze-dried, replaced with nitrogen, and stoppered and wound to obtain a dissolution-type dry preparation for use (control product 4).

(5) Elcatonin (made by Asahi Kasei Corporation)
1.5 mg and dextran 500 mg were taken, and 50 ml of sterile distilled water was added and dissolved. After aseptic filtration, 0.5 ml each was dispensed into a glass vial, freeze-dried, replaced with nitrogen, and stoppered and wound to obtain a dissolution-type dry preparation before use (control product 5).

(6) Elcatonin (made by Asahi Kasei Corporation)
1.5 mg, lactose 1250 mg and sodium chloride 450 mg were taken, and sterile distilled water 50 ml was added and dissolved. After aseptic filtration, dispense 0.5 ml each into a glass vial, freeze-dry and replace with nitrogen, then stopper and wrap.
A dry formulation that was dissolved before use was obtained (control product 6).

Test method

The products 1 to 6 of the present invention obtained in the above-mentioned Examples 1 to 6
Each of Control Products 1 to 6 was stored at 40 ° C., sampled over time, and the elcatonin content was measured using high performance liquid chromatography (HPLC) under the following conditions. HPLC measurement condition column: YMC AM-302 ODS S-512
0 (manufactured by YMC), inner diameter 4.6 × 150 mm, mobile phase: 0.1% TFA: acetonitrile (65: 3)
5) Flow rate: 1.0 ml / min Detection: UV 220 nm

Results Table 2 shows the composition of each sample and the residual ratio of elcatonin stored at 40 ° C for 3 months.

[0032]

[Table 2]

As shown in Table 2, the saccharide alone has a temperature of 40 °
C, the stability after 3 months decreased, and in the sample containing sodium chloride alone or a large amount of sodium chloride added, the cake contracted during freeze-drying, which was not preferable, and the stability also decreased. On the other hand, all of the products of the present invention in which sodium chloride and saccharides were used in combination had a stable residual activity of calcitonin of 85% or more after 3 months at 40 ° C, and the combined effect was recognized. As described above, by using sugar and a certain amount of sodium chloride as a stabilizer for calcitonins, it became possible to obtain a stable and highly safe lyophilized preparation of calcitonins.

Continuation of front page (51) Int.Cl. ⁷ identification code FI A61P 19/10 A61K 37/30 (58) Fields investigated (Int.Cl. ⁷ , DB name) A61K 38/23 A61K 47/02 A61K 47 / 26 A61P 3/14 A61P 19/08 A61P 19/10 CA (STN) REGISTRY (STN) JISST file (JOIS)

Claims (7)

(57) [Claims] 1. Elcatonin as an active ingredient, containing a saccharide and sodium chloride, wherein the amount of sodium chloride is 1/100 to 1/4 parts by weight per 1 part by weight of the saccharide.
And a freeze-dried composition. 2. The saccharide is one or more saccharides selected from the group consisting of mannitol, glucose, sorbitol, inositol, xylitol, galactose, fructose, sucrose, maltose, lactose, trehalose, dextran, cyclodextrin. The freeze-dried composition according to claim 1. 3. The freeze-dried composition is a freeze-dried preparation.
The freeze-dried composition according to claim 1 or 2,
object. 4. The freeze-dried composition is an injection.
The freeze-dried composition according to any one of claims 1 to 3.
object. 5. The freeze-dried composition is in a cake state.
Freeze-drying according to any one of claims 1 to 4.
Composition. 6. A single saccharide containing elcatonin as an active ingredient.
Sodium chloride 1/100 to 1/4 parts by weight to parts by weight
Lyophilization of elcatonin, which is characterized in that the saccharide and sodium chloride are dissolved in an aqueous medium and then lyophilized.
A method for stabilizing the storage stability of a dried composition . 7. The saccharide is one or more saccharides selected from the group consisting of mannitol, glucose, sorbitol, inositol, xylitol, galactose, fructose, sucrose, maltose, lactose, trehalose, dextran, cyclodextrin. The stabilization method according to claim 6 .