JP3608802B2 - Stable calcitonin pharmaceutical composition and method for producing the same - Google Patents

Description

表１に示すごとく、実施例では比較例及び対照例と比べて安定であった。
【００４１】
試験例２．
実施例７〜１１及び比較例９、１０で調製した試料溶液を、室温で振幅３ｃｍ、１００回／分の振盪回数で振盪し、外観（溶状）を観察した。また、溶液のヒトカルシトニン濃度を液体クロマトグラフィーで定量し、ヒトカルシトニンの残存率を求めた。その結果を表２に示す。
【００４２】

表２に示すごとく、実施例では比較例及び対照例と比べて安定であった。
【００４３】
試験例３．
実施例７〜９で調製した試料溶液を、室温で振幅３ｃｍ、１００回／分の振盪回数で７日間振盪し、その後の溶液の生物活性を以下のように検討した。即ち、２４時間絶食後、引き続き２４時間低カルシウム食で飼育したＳ．Ｄ．系雄性ラット（６週齢）を、ペントバルビタール麻酔下、背位固定し、大腿動脈カニューレ及び気管カニューレを施した。試料溶液５μｌ（ヒトカルシトニン５０μｇ／ｋｇ）を鼻腔内に投与し、経時的に大腿動脈より採血し、血清カルシウム濃度をカルシウム測定用試薬（サンアッセイＣａ：三光純薬社製）で定量した。結果を表３に示す。
【００４４】

表３に示すごとく、７日間振盪させた溶液においても血清カルシウム濃度の低下が認められた。
【００４５】
【発明の効果】
以上、本発明により、局所刺激性が少なく、実用的でかつ溶液で安定なカルシトニン溶液を提供するカルシトニン医薬組成物を製造することができる。[0001]
[Industrial application fields]
The present invention relates to a stable calcitonin pharmaceutical composition and a method for producing the same.
[0002]
[Prior Art and Problems to be Solved by the Invention]
Calcitonin is a polypeptide hormone having various pharmacological activities used for the treatment of pain improvement, hypercalcemia, Paget's disease and the like in osteoporosis. Although calcitonin is usually administered by injection, depending on the disease, long-term continuous administration is required, and a simple administration formulation other than injection is desired. Nasal administration is one of the methods, but in this case, it is desirable that a single dose is 0.2 to 0.3 ml or less, and if the liquid volume is large, it leaks from the nasal cavity and cannot be effectively administered.
[0003]
Therefore, in order to administer an effective therapeutic amount, a relatively high concentration of drug solution is required. For example, for human calcitonin, it is desirable to prepare a drug solution of 0.5 mg / ml or more. However, calcitonin easily aggregates in a solution, and easily aggregates due to mechanical stress such as shaking. Moreover, the higher the concentration, the more unstable, and it is difficult to stably prepare a solution having a calcitonin concentration desirable for treatment.
[0004]
If the drug solution causes a cloudy precipitate due to aggregation, uniform administration is impossible, and absorption from the nasal mucosa is significantly inhibited. In general, gelatin, albumin, some surfactants, etc. have been reported to prevent aggregation of proteins and peptides. However, a practical and stable calcitonin pharmaceutical composition and calcitonin solution should be prepared with little local irritation. Was impossible.
[0005]
[Means for Solving the Problems]
As a result of intensive studies on the stabilization of calcitonin solutions by various additives that are acceptable for medical use, the present inventors have formulated a certain kind of additive to provide a stable and absorption at a concentration desired for treatment. The present inventors have found that a good calcitonin pharmaceutical composition and a calcitonin solution can be prepared.
[0006]
That is, the present invention relates to a stable calcitonin pharmaceutical composition and a method for producing a calcitonin solution, and as a stabilizer, citric acid, polyvinyl alcohol, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, It is characterized by containing a plurality.
[0007]
These stabilizers (especially citric acid) are effective even when used alone, but preferably (1) citric acid, and (2) polyvinyl alcohol, polyoxyethylene sorbitan fatty acid ester and polyoxyethylene It is desirable to contain any one or more of hydrogenated castor oil, and more preferably, among these combinations, for example, citric acid and polyvinyl alcohol, citric acid and polyoxyethylene sorbitan fatty acid ester, or citric acid and polyoxyethylene It is desirable to combine hardened castor oil. By adding these stabilizers to the calcitonin pharmaceutical composition or calcitonin solution, a further stabilizing effect is recognized.
[0008]
The citric acid used in the present invention is added and mixed so as to be usually 0.05 to 5% (W / V), preferably 0.1 to 2% (W / V) with respect to the chemical solution.
Polyvinyl alcohol is a synthetic polymer obtained by radical polymerization of vinyl acetate, and is usually 0.001 to 10% (W / V), preferably 0.01 to 5% (W / V) with respect to the chemical solution. Add and mix so that
Polyoxyethylene sorbitan fatty acid ester is a nonionic surfactant obtained by addition polymerization of ethylene oxide to sorbitan fatty acid ester, and examples thereof include polysorbate-40, polysorbate-60, and polysorbate-80.
[0009]
The polyoxyethylene sorbitan fatty acid ester is added and mixed so as to be usually 0.001 to 1% (W / V), preferably 0.005 to 0.5% (W / V) with respect to the chemical solution.
Polyoxyethylene hydrogenated castor oil is a nonionic surfactant obtained by addition polymerization of ethylene oxide to a hydrogenated oil obtained by adding hydrogen to castor oil. For example, Nikkor HCO-10, Nikkor HCO-40, Nikkor HCO-50, Nikkor HCO-60, etc. are mentioned.
The polyoxyethylene hydrogenated castor oil is added to and mixed with the chemical solution so that it is usually 0.001 to 1% (W / V), preferably 0.005 to 0.5% (W / V).
[0010]
The pharmaceutical composition of the present invention can be in the form of a solution or in the form of a powder to be used after being dissolved in a specific solution. In the case of a powder used by dissolving in a solution at the time of use, citric acid, polyvinyl alcohol, polyoxyethylene sorbitan fatty acid ester and / or polyoxyethylene hydrogenated castor oil may be added to either the powder or the solution, Moreover, you may add to both a powder and a solution.
[0011]
As additives, pH adjusters, bactericides / preservatives, viscosity modifiers, freeze-drying aids, absorption accelerators and the like can be added as necessary.
As the pH adjuster, hydrochloric acid, acetic acid or the like is used.
Any bactericidal / preservative may be used as long as it is usually used in pharmaceutical compositions, and examples thereof include paraoxybenzoic acid esters and benzalkonium chloride.
Examples of the viscosity modifier include polyvinyl pyrrolidone and hydroxypropyl methylcellulose.
[0012]
Examples of the lyophilization aid include aminoacetic acid, mannitol, sucrose, glucose, dextran and the like.
Examples of absorption promoters include sodium caprate, bestatin, amastatin, nafamostat mesylate, camostat mesylate, aprotinin and the like.
[0013]
As a dosage form of the pharmaceutical composition of the present invention, a solution or a drug solution obtained by dissolution at the time of use can be administered by dripping or spraying using a drop container for nasal drop, a spray container or a nasal aerosol applicator. .
The calcitonin that can be used in the present invention is not particularly limited since those derived from humans, pigs, cows, salmon, eels, rats and the like can be used, but in the following examples, examples using human calcitonin are shown. .
[0014]
【Example】
EXAMPLES The present invention will be specifically described below with reference to examples and test examples, but the present invention is not limited to these.
Example 1.
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 15mg
Purified water was added to a final volume of 10 ml.
[0015]
Example 2
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Polyvinyl alcohol 100mg
Purified water was added to a final volume of 10 ml.
[0016]
Example 3
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Polysorbate 80 10mg
Purified water was added to a final volume of 10 ml.
[0017]
Example 4
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Citric acid 15mg
Purified water was added to a final volume of 10 ml.
[0018]
Example 5 FIG.
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Polyvinyl alcohol 100mg
Purified water was added to a final volume of 10 ml.
[0019]
Example 6
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Polysorbate 80 10mg
Purified water was added to a final volume of 10 ml.
[0020]
Example 7
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 60mg
Polyvinyl alcohol 100mg
Purified water was added to a final volume of 10 ml.
[0021]
Example 8 FIG.
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 60mg
Polysorbate 80 1mg
Purified water was added to a final volume of 10 ml.
[0022]
Example 9
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 60mg
Polyoxyethylene hydrogenated castor oil ^{* 1} 1mg
Purified water was added to a final volume of 10 ml.
* 1: Nikkor HCO-60 (Nikko Chemicals Corporation)
[0023]
Example 10
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 60mg
Polyvinyl alcohol 100mg
Aprotinin 1mg
Purified water was added to a final volume of 10 ml.
[0024]
Example 11
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 60mg
Polysorbate 80 1mg
Nafamostat mesylate 1mg
Purified water was added to a final volume of 10 ml.
[0025]
Example 12
20 mg of human calcitonin and 1000 mg of purified sucrose were dissolved in 10 ml of purified water and filtered aseptically. The filtrate was aseptically filled into vials and lyophilized. As an attached solution, 2 mg of benzalkonium chloride and 15 mg of citric acid were dissolved in 10 ml of purified water, and a sterile filtered solution was prepared.
[0026]
Control Example 1
Human calcitonin 20mg
Purified water was added to a final volume of 10 ml.
[0027]
Control Example 2.
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Purified water was added to a final volume of 10 ml.
[0028]
Control Example 3.
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Purified water was added to a final volume of 10 ml.
[0029]
Comparative Example 1
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Taurocholic acid 100mg
Purified water was added to a final volume of 10 ml.
[0030]
Comparative Example 2
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Deoxycholic acid 100mg
Purified water was added to a final volume of 10 ml.
[0031]
Comparative Example 3
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Sucrose fatty acid ester ^{* 2} 125mg
Purified water was added to a final volume of 10 ml.
* 2: Ryoto Sugar Ester S1670 (Mitsubishi Kasei Foods)
[0032]
Comparative Example 4
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Sucrose fatty acid ester ^{* 3} 125mg
Purified water was added to a final volume of 10 ml.
* 3: Ryoto sugar ester P1670 (Mitsubishi Kasei Foods)
[0033]
Comparative Example 5
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Taurocholic acid 100mg
Purified water was added to a final volume of 10 ml.
[0034]
Comparative Example 6
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Deoxycholic acid 100mg
Purified water was added to a final volume of 10 ml.
[0035]
Comparative Example 7
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Sucrose fatty acid ester ^{* 2} 125mg
Purified water was added to a final volume of 10 ml.
* 2: Ryoto Sugar Ester S1670 (Mitsubishi Kasei Foods)
[0036]
Comparative Example 8
Human calcitonin 20mg
Mannitol 500mg
Benzalkonium chloride 2mg
Sucrose fatty acid ester ^{* 3} 125mg
Purified water was added to a final volume of 10 ml.
* 3: Ryoto sugar ester P1670 (Mitsubishi Kasei Foods)
[0037]
Comparative Example 9
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 60mg
Human serum albumin 200mg
Purified water was added to a final volume of 10 ml.
[0038]
Comparative Example 10
Human calcitonin 20mg
Refined white sugar 1000mg
Benzalkonium chloride 2mg
Citric acid 60mg
Gelatin 100mg
Purified water was added to a final volume of 10 ml.
The said calcitonin pharmaceutical composition in this invention is very stable as shown in the following test examples 1-3.
[0039]
Test Example 1
The sample solutions prepared in Examples 1 to 6, Comparative Examples 1 to 8, and Control Examples 1 to 3 were shaken at room temperature with an amplitude of 3 cm and a shaking frequency of 100 times / minute, and the appearance (solution state) was observed. The results are shown in Table 1.
[0040]

As shown in Table 1, the examples were more stable than the comparative examples and the control examples.
[0041]
Test Example 2
The sample solutions prepared in Examples 7 to 11 and Comparative Examples 9 and 10 were shaken at room temperature with an amplitude of 3 cm and a shaking frequency of 100 times / minute, and the appearance (solution state) was observed. Further, the concentration of human calcitonin in the solution was quantified by liquid chromatography, and the residual ratio of human calcitonin was determined. The results are shown in Table 2.
[0042]

As shown in Table 2, the example was more stable than the comparative example and the control example.
[0043]
Test Example 3
The sample solutions prepared in Examples 7 to 9 were shaken at room temperature with an amplitude of 3 cm and a shaking frequency of 100 times / minute for 7 days, and then the biological activity of the solutions was examined as follows. That is, after 24 hours fasting, S. bred was kept on a low calcium diet for 24 hours. D. Strain male rats (6 weeks old) were fixed in the dorsal position under pentobarbital anesthesia and subjected to femoral artery cannula and tracheal cannula. 5 μl of sample solution (human calcitonin 50 μg / kg) was administered intranasally, blood was collected from the femoral artery over time, and the serum calcium concentration was quantified with a calcium measurement reagent (Sun Assay Ca: manufactured by Sanko Junyaku Co., Ltd.). The results are shown in Table 3.
[0044]

As shown in Table 3, a decrease in serum calcium concentration was also observed in the solution shaken for 7 days.
[0045]
【The invention's effect】
As described above, according to the present invention, it is possible to produce a calcitonin pharmaceutical composition that provides a calcitonin solution that is practical and stable with little local irritation.

Claims (3)

In a liquid preparation containing calcitonin at a concentration of 0.5 mg / mL or more, calcitonin can be added by adding one or more aggregation inhibitors of citric acid, polyvinyl alcohol, polyoxyethylene sorbitan fatty acid ester and polyoxyethylene hydrogenated castor oil. how to suppress aggregation. The concentration of the aggregation inhibitor in the liquid is 0.05 to 5% (W / V) for citric acid, 0.001 to 10% (W / V) for polyvinyl alcohol, and 0.001 for polyoxyethylene sorbitan fatty acid ester. The method of claim 1, which is ˜1% (W / V), and 0.001 to 1% (W / V) for polyoxyethylene hydrogenated castor oil. The method according to claim 1 or 2, which is a nasal agent.