JPH06786B2 - Method for producing crystalline freeze-dried preparation of sodium cefarotin - Google Patents

Description

【発明の詳細な説明】 本発明はセファロスポリン系抗生物質であるセファロチ
ンナトリウムの結晶性凍結乾燥製剤、特に顆粒状品の製
造法に関するものである。The present invention relates to a method for producing a crystalline lyophilized preparation of cephalosporin sodium, which is a cephalosporin antibiotic, in particular a granular product.

周知の如くセファロチンナトリウムは各種の疾病の予防
および／または治療に広く用いられている重要な医薬品
であり、従来その注射用製剤は、普通セファロチンナト
リウムの結晶性粉末を注射用バイアル（瓶）に小分けす
る粉末充填法により製造されてきたが、この方法では微
小な異物の混入を避け得ないので、近時凍結乾燥法が採
用されることが多い。As is well known, cefarotin sodium is an important drug that is widely used for the prevention and / or treatment of various diseases. Conventionally, the injectable preparation is usually a crystalline powder of cephalotin sodium for injection vials (bottles). It has been manufactured by the powder filling method divided into small parts. However, since this method cannot avoid the inclusion of minute foreign matter, the freeze-drying method is often adopted recently.

しかしながら、通常の凍結乾燥法により得られるセファ
ロチンナトリウムは無定形（アモルファス）の乾燥粉末
であって安定性に欠け、比較的短期間の間に着色するこ
とが知られている。However, it is known that cephalothin sodium obtained by a usual freeze-drying method is an amorphous (amorphous) dry powder, lacks stability, and is colored in a relatively short period of time.

従って、セファロチンナトリウムの凍結乾燥に際して、
これを安定な、そして製品価値を損なうことのない結晶
の形で得ようとするセファロチンナトリウムの改良され
た凍結乾燥法も既にいくつか提案されている。Therefore, during freeze-drying of cephalotin sodium,
Several improved freeze-drying processes of sodium cephalotin have already been proposed in an attempt to obtain this in the form of crystals that are stable and do not impair product value.

すなわち、 (1)セファロチンナトリウムの水溶液を予備凍結し、次
いで昇温して結晶を析出させた後、水を昇華させる方法
（特開昭５１−１２３８１３号）。That is, (1) a method in which an aqueous solution of sodium cephalothin is pre-frozen, then the temperature is raised to precipitate crystals, and then water is sublimated (JP-A-51-123813).

(2)有機溶媒を含むセファロチンナトリウムの水溶液を
予備凍結し、次いで昇温させてセファロチンナトリウム
の結晶を析出させた後、凍結乾燥を行う方法（特開昭５
６−１２０６１５号）。(2) A method of pre-freezing an aqueous solution of sodium cephalotin containing an organic solvent, then heating the temperature to precipitate crystals of sodium cephalothin, and then performing freeze-drying (Japanese Patent Laid-Open No. Sho 5).
6-120615).

(3)セファロチンナトリウムの飽和もしくは過飽和水溶
液にセファロチンナトリウムの微細結晶（種晶）を添加
し、この液を予備凍結した後、昇温して結晶を析出させ
凍結乾燥する、という本出願自身の開発にかかる方法
（特願昭５８−３６４６１号（特開昭６０−１１４９１
号））。(3) The present application itself, in which fine crystals (seed crystals) of sodium cephalotin are added to a saturated or supersaturated aqueous solution of sodium cephalotin, the liquid is pre-frozen, and then heated to precipitate crystals and freeze-dry. (Japanese Patent Application No. 58-36461 (JP-A-60-11491)
issue)).

などである。And so on.

しかしながら、上記の改良法でも、その(1)、(2)の方法
は、結晶の析出に長時間を要し、また微量のアモルファ
ス分の形成を抑制し得ず、経時的な着色も避けられない
という欠点がある。一方、上記(3)の方法では、短時間
での結晶析出および経時的な着色の防止も共に可能であ
るが、その反面セファロチンナトリウムの種晶を無塵、
無菌的に調製し、添加しなければならないという煩雑さ
を伴っている。However, even in the above improved method, the methods (1) and (2) require a long time for the precipitation of crystals, cannot suppress the formation of a trace amount of amorphous components, and avoid coloring over time. It has the drawback of not having it. On the other hand, in the above method (3), it is possible to prevent crystal precipitation in a short time and coloration over time, but on the other hand, dust-free seed crystals of cephalothin sodium,
It involves the complexity of aseptically preparing and adding.

このような状況に鑑み、本発明者等は、特別な添加物を
使用せずにセファロチンナトリウムの結晶性粉末を得る
ための凍結乾燥法について鋭意研究した結果、凍結乾燥
に付すセファロチンナトリウム水溶液の濃度、保存条
件、予備凍結速度および予備凍結後の再昇温々度（晶析
温度）を特定の範囲にコントロールすることによって著
しく晶析時間を短縮することができ、ほぼ完全にアモル
ファス分の形成を抑制し得、しかもこのとき意外にも乾
燥速度および再溶解速度のきわめて速い顆粒状の結晶性
凍結乾燥品が得られることを見い出し、本発明を完成す
るに至った。In view of such a situation, the present inventors have diligently studied a freeze-drying method for obtaining a crystalline powder of cefarotin sodium without using a special additive, and a cefalotin sodium aqueous solution to be freeze-dried. The crystallization time can be significantly shortened by controlling the concentration, storage conditions, pre-freezing rate, and reheating temperature (crystallization temperature) after pre-freezing within a specific range. It has been found that a granular crystalline freeze-dried product which can suppress the above-mentioned phenomenon and which has a surprisingly high drying rate and re-dissolution rate can be obtained at this time, thus completing the present invention.

すなわち、本発明の要旨は、水溶液の濃度が２６〜３６
重量％であり、かつ、その温度が１０〜３０℃であるセ
ファロチンナトリウム過飽和水溶液を、０．５℃／分以
下の冷却温度で−１５℃以下に徐冷凍結後、−５〜−２
℃まで昇温せしめて晶析を行い、次いで−３０〜−４０
℃に再冷却後凍結乾燥工程に付すことに存し、セファロ
チンナトリウムの顆粒状の結晶性凍結乾燥品を得ること
をその目的とするものである。That is, the gist of the present invention is that the concentration of the aqueous solution is 26 to 36.
The supersaturated aqueous solution of sodium cephalotin, which is 10% to 30 ° C by weight, is slowly cooled to -15 ° C or less at a cooling temperature of 0.5 ° C / min or less, and then -5 to -2.
Crystallization is performed by raising the temperature to ℃, and then -30 to -40
The purpose is to obtain a granular crystalline freeze-dried product of cephalotin sodium, which consists in subjecting it to a freeze-drying step after recooling to ° C.

以下に本発明方法の要点について詳述する。The essential points of the method of the present invention will be described in detail below.

まず、本発明においては、凍結乾燥に付すセファロチン
ナトリウムの水溶液は過飽和水溶液でなければならな
い。セファロチンナトリウムの３０℃における水への溶
解度は２４．５％（重量％、以下同じ）であるが、目的
とする顆粒状の晶析を効率よく行うためには２６％以上
の水溶液濃度とすることが好ましい。しかし、過飽和度
が過ぎると以後無菌濾過を行う時にセファロチンナトリ
ウムの結晶析出を生じ易くなるので、３６％を上限と
し、最も好ましい濃度範囲は２８〜３２％である。First, in the present invention, the aqueous solution of cephalotin sodium to be freeze-dried must be a supersaturated aqueous solution. The solubility of cephalotin sodium in water at 30 ° C. is 24.5% (weight%, the same applies hereinafter), but in order to efficiently carry out the desired granular crystallization, the aqueous solution concentration is 26% or more. It is preferable. However, when the degree of supersaturation is too high, crystal precipitation of sodium cephalotin is likely to occur during subsequent sterile filtration, so the upper limit is 36%, and the most preferable concentration range is 28 to 32%.

次に、上記のセファロチンナトリウム水溶液を１０〜３
０℃に約１時間保持する。このときの温度が１０℃未満
の場合には後述する晶析工程で結晶化度の小さい稠密複
晶が生成し易くなり、また３０℃を越えると同じく晶析
工程ではセファロチンナトリウムの結晶成長が著しく阻
害されるため、いずれの場合も顆粒状の結晶性凍結乾燥
品の生成率が著しく低下する。従って、より好ましい温
度範囲は１５〜２６℃である。Next, 10 to 3 of the above-mentioned aqueous solution of sodium cephalothin is added.
Hold at 0 ° C. for about 1 hour. If the temperature at this time is lower than 10 ° C., a dense double crystal having a low degree of crystallinity is likely to be generated in the crystallization step described later, and if it exceeds 30 ° C., the crystal growth of sodium cephalotin also occurs in the crystallization step. In both cases, the production rate of the granular crystalline freeze-dried product is remarkably reduced because of significant inhibition. Therefore, the more preferable temperature range is 15 to 26 ° C.

次いで、至適温度に保持した上記セファロチンナトリウ
ム水溶液をバイアルに所定量分注（充填）する。バイア
ルは適当な容量のものでよいが、例えば１４mlバイアル
の場合はセファロチンナトリウム１ｇ力価相当量、１０
０mlバイアルの場合は１ｇ〜２ｇ力価相当量の溶液を分
注する。Next, a predetermined amount of the above-mentioned aqueous solution of cephalotin sodium maintained at the optimum temperature is dispensed (filled) into a vial. The vial may have an appropriate volume, but for example, in the case of a 14 ml vial, 1 g of cephalotin sodium has a titer equivalent to 10,
For a 0 ml vial, dispense 1 g to 2 g of titer equivalent solution.

上記バイアルを１０〜３０℃の任意の温度に設定した棚
に搬入した後、０．５℃／分以下の冷却速度、好ましく
は０．０５〜０．５℃／分の冷却温度で−１５℃、より
好ましくは−３０℃以下に冷却し、バイアル内溶液を凍
結する。このとき冷却速度が０．５℃／分を越えると凍
結した内容液の氷晶が小さくなり過ぎ、その結果セファ
ロチンナトリウムを含むスケルトン部の厚みが薄くなる
ために晶析工程での顆粒成長が阻害される。また、冷却
速度が０．０５℃／分未満ではバイアル内容液の完全凍
結までの所要時間が長過ぎて好ましくない。それ故最も
好ましい冷却速度の範囲は０．１〜０．３℃／分であ
る。所要凍結時間は約２時間であるが、徐冷後約１時間
放置することが好ましい。After carrying the above vial into a shelf set to an arbitrary temperature of 10 to 30 ° C., a cooling rate of 0.5 ° C./min or less, preferably −15 ° C. at a cooling temperature of 0.05 to 0.5 ° C./min. More preferably, the solution in the vial is frozen by cooling to −30 ° C. or lower. At this time, if the cooling rate exceeds 0.5 ° C./minute, the ice crystals of the frozen content liquid become too small, and as a result, the skeleton portion containing sodium cephalotin becomes thin, so that the grain growth in the crystallization step occurs. Be hindered. Further, if the cooling rate is less than 0.05 ° C./minute, the time required for completely freezing the vial content liquid is too long, which is not preferable. Therefore, the most preferable cooling rate range is 0.1 to 0.3 ° C./min. The required freezing time is about 2 hours, but it is preferable to leave it for about 1 hour after gradual cooling.

次いで、棚温をセファロチンナトリウム水溶液の融解終
了温度（−１℃）よりおよそ４〜１℃低い任意の温度に
まで昇温し、一定時間その温度に保持する。この間にセ
ファロチンナトリウムの針状晶が氷晶界面付近を起点と
して放射状に成長する。晶析のために棚温を一定に保つ
時間、すなわち晶析時間は少なくとも１時間であり、好
ましくは２〜３時間である。また、晶析温度が低すぎる
と結晶化度の小さい稠密複晶が生じ易く、晶析温度が高
すぎると結晶成長が著しく阻害され、アモルファス分が
形成し易くなることから晶析温度の好ましい範囲は−
４．５〜−３．５℃である。Then, the shelf temperature is raised to an arbitrary temperature which is lower by about 4 to 1 ° C. than the melting end temperature (-1 ° C.) of the sodium cephalotin aqueous solution, and the temperature is maintained for a certain period of time. During this period, acicular needles of sodium cephalotin grow radially starting from the vicinity of the ice crystal interface. The time for keeping the shelf temperature constant for crystallization, that is, the crystallization time is at least 1 hour, preferably 2-3 hours. Further, if the crystallization temperature is too low, a dense double crystal with a small degree of crystallinity is likely to occur, and if the crystallization temperature is too high, crystal growth is significantly inhibited, and an amorphous component is easily formed. Is-
It is 4.5 to -3.5 ° C.

上記の晶析操作が完了したらバイアルを−３０℃以下に
再冷却した後、常法により凍結乾燥を行う。例えば、セ
ファロチンナトリウム１ｇ力価相当量を分注したバイア
ルの場合は、棚温３０℃、真空度０．２mbarで５〜１５
時間、次いで棚温５０℃、真空度０．２mbarで１〜３時
間乾燥を行う。After the above crystallization operation is completed, the vial is recooled to -30 ° C or lower, and then freeze-dried by a conventional method. For example, in the case of a vial in which 1 g of cefalotin sodium titer is dispensed, 5 to 15 at a shelf temperature of 30 ° C and a vacuum degree of 0.2 mbar.
Drying is carried out for 1 hour at a shelf temperature of 50 ° C. and a vacuum degree of 0.2 mbar for 1 hour.

以上のような一連の操作を連続して実施することにより
バイアルに充填されたセファロチンナトリウムの粒径
０．２〜１．５mmの顆粒状の結晶性凍結乾燥品が得られ
る。By continuously carrying out the above-mentioned series of operations, a granular crystalline lyophilized product having a particle size of 0.2 to 1.5 mm of cefarotin sodium filled in a vial can be obtained.

上記本発明によれば、以下の実験にも示すように凍結乾
燥時間を従来法に比して著しく短縮することができ、ま
た、再溶解速度が速く、結晶化度も大きく、さらに経時
的着色度のきわめて低い製品的価値に優れた顆粒状の結
晶性凍結乾燥品を得ることができる。According to the present invention, the freeze-drying time can be significantly shortened as compared with the conventional method as shown in the following experiment, the re-dissolution rate is fast, the crystallinity is large, and the coloring over time is further achieved. It is possible to obtain a granular crystalline lyophilized product which has an extremely low product value and is excellent.

実験１（凍結乾燥品の製法と諸物性との関係） 本発明方法で製造したセファロチンナトリウムの凍結乾
燥品の結晶化度、外観、再溶解時間、経時的着色度の増
加および含量変化について、それぞれ従来法で得たもの
と比較した。Experiment 1 (Relationship between production method of freeze-dried product and various physical properties) With respect to crystallinity, appearance, redissolution time, increase in coloring degree with time and content change of the freeze-dried product of cephalotin sodium produced by the method of the present invention, Each was compared with that obtained by the conventional method.

（ａ）凍結乾燥品の製造法と結晶化度との関係 本発明方法に従う後記実施例１，２および３で得られた
凍結乾燥品Ａ、ＢおよびＣ、特開昭５１−１２３８１３
号公報に開示されている製造法（参考例１）で得られた
凍結乾燥品Ｄ、および特願昭５８−３６４６１号に開示
した製造法（参考例２）により得られた凍結乾燥品Ｅの
結晶化度を熱分析法により測定した。結果を表１に示
す。なお、凍結乾燥品Ｄの晶析時間は１５時間である。(A) Relationship between production method of freeze-dried product and crystallinity Freeze-dried products A, B and C obtained in Examples 1, 2 and 3 described below according to the method of the present invention, JP-A-51-123813.
Of the freeze-dried product D obtained by the production method disclosed in Japanese Patent Publication (Reference Example 1) and the freeze-dried product E obtained by the production method disclosed in Japanese Patent Application No. 58-36461 (Reference Example 2). The crystallinity was measured by thermal analysis. The results are shown in Table 1. The crystallization time of freeze-dried product D is 15 hours.

上表から明らかなように本発明に従って製造した試料
Ａ、ＢおよびＣの結晶化度はいずれも１００％で、種晶
を添加して製造した場合のＥと等しく良好な結晶化度を
示し、種晶を用いない公知の方法で製造したＤの試料と
比較すると有意に優れていることが分る。 As is clear from the above table, the crystallinity of each of the samples A, B and C produced according to the present invention was 100%, and showed a crystallinity as good as E when produced by adding a seed crystal, It can be seen that it is significantly superior to the sample of D produced by a known method without using seed crystals.

（ｂ）凍結乾燥品の製造法と製品外観との関係 上記（ａ）で使用した凍結乾燥品の外観をそれぞれ観察
してみると、本発明品である試料Ａ、Ｂ、Ｃは直径約
０．３mmの顆粒の集合体であり、他の試料Ｄ、Ｅの結晶
性粉末ケーキとは全く異なっている。(B) Relationship between production method of freeze-dried product and product appearance When the appearance of the freeze-dried product used in (a) above is observed, Samples A, B, and C of the present invention have a diameter of about 0. It is an aggregate of 0.3 mm granules, which is completely different from the other crystalline powder cakes of Samples D and E.

（ｃ）凍結乾燥品の製造法と再溶解時間との関係 上記（ｂ）と同じ試料Ａ、Ｄ、Ｅに、それぞれ２５℃の
蒸留水４mlを入れ、手振盪により再溶解させたときの完
全溶解までの所要時間を測定した。(C) Relationship between production method of freeze-dried product and redissolution time 4 ml of distilled water at 25 ° C. was added to each of the same samples A, D, and E as in (b) above, and they were completely dissolved by hand shaking. The time required for dissolution was measured.

結果を表２に示す。The results are shown in Table 2.

上表から明らかなように本発明により得た試料Ａ（１ｇ
力価）の再溶解時間は、他の方法によって製造した結晶
性凍結乾燥ケーキであるＤ、Ｅのそれに比べて著しく短
いことが分かる。これは上記（ｂ）でも明らかなように
結晶性顆粒を形成している試料Ａの溶解に対する実効表
面積が試料Ｄ、Ｅに比し著しく増大していることに起因
する。なお、試料Ｄの再溶解時間がＥよりもかなり長い
のは、種晶添加法によって得た試料Ｅに比べ種晶無添加
のＤには結晶化度の小さい稠密複晶が多く含まれている
ためである。 As is apparent from the above table, sample A (1 g
It can be seen that the re-dissolution time of (titer) is significantly shorter than that of crystalline freeze-dried cakes D, E produced by other methods. This is because the effective surface area for dissolution of the sample A forming crystalline granules is remarkably increased as compared with the samples D and E, as is apparent from the above (b). The re-dissolution time of Sample D is considerably longer than that of E. Compared to Sample E obtained by the seed crystal addition method, D without seed crystal addition contains a large amount of dense double crystals with a small crystallinity. This is because.

（ｄ）凍結乾燥品の製造法と経時的着色度との関係 上記（ｃ）と同じ試料について、５０℃におけるそれぞ
れの経時的着色度を調べた。着色度は各試料の２０％再
溶解水溶液の４１０nmの吸光度を測定することにより決
定し、その吸光度の増加をΔＥ₄₁₀で表わした。結果を
第１図に示す 第１図より試料Ａは、種晶添加法により結晶化度を１０
０％とした試料Ｅと同様に、結晶化度のやや小さい試料
Ｄに比べて着色変化が著しく少ないことが分かる。(D) Relationship between production method of freeze-dried product and coloring over time With respect to the same sample as (c) above, the coloring over time at 50 ° C. was examined. The degree of coloration was determined by measuring the absorbance at 410 nm of a 20% redissolved aqueous solution of each sample, and the increase in the absorbance was represented by ΔE ₄₁₀ . The results are shown in FIG. 1. As shown in FIG. 1, Sample A has a crystallinity of 10 by the seed crystal addition method.
It can be seen that, similarly to the sample E with 0%, the change in color is significantly smaller than that of the sample D having a slightly small crystallinity.

（ｅ）凍結乾燥品の製造法と、経時的含量変化との関係 上記（ｄ）と同じ試料につき、５０℃におけるセファロ
チンナトリウムの経時的な含量変化を高速液体クロマト
グラフィー（ＨＰＬＣ）により調べた。その結果は第２
図に示すとおりいずれの試料も含量変化はなかった。(E) Relationship between production method of freeze-dried product and change in content over time With respect to the same sample as in (d) above, change in content of sodium cephalotin at 50 ° C with time was examined by high performance liquid chromatography (HPLC). . The result is the second
As shown in the figure, there was no change in the content of any of the samples.

実験２（凍結乾燥品の製造法と凍結乾燥速度との関係） 上記実験１（ｄ）と同じ試料Ａ、ＤおよびＥについて凍
結乾燥時の乾燥速度を凍乾庫内蔵天秤により測定した。
なお、このときの乾燥条件は一度−４０℃まで冷却した
後、棚温３０℃、真空度０．４mbarで一定とした。結果
を下記表３に示す。Experiment 2 (Relationship between Freeze-Dried Product Manufacturing Method and Freeze-Drying Rate) For the same samples A, D, and E as in the above-mentioned Experiment 1 (d), the drying rate at the time of freeze-drying was measured by a freeze-dryer built-in balance.
The drying conditions at this time were such that the temperature was once cooled to −40 ° C., then the shelf temperature was 30 ° C., and the vacuum degree was 0.4 mbar. The results are shown in Table 3 below.

表３には単位昇華表面積に対する１次乾燥初速度が示さ
れている。これにより明らかなように顆粒状晶析を行っ
た本発明品の試料Ａは、結晶性セーキを形成する試料Ｄ
およびＥに比べて凍結乾燥速度は大きく、該表の最下欄
に示す同一乾燥条件での蒸留水の乾燥速度と比べてほぼ
等しいことが分かる。なお、試料Ｄの乾燥速度がＥに比
べて小さいのは前記実験１の（ｃ）の場合と同じ理由に
よるものである。 Table 3 shows the primary drying initial rate per unit sublimation surface area. As apparent from this, the sample A of the product of the present invention in which the granular crystallization was performed is the sample D which forms a crystalline shake.
It can be seen that the freeze-drying rate is higher than those of E and E, and is almost equal to the drying rate of distilled water under the same drying conditions shown in the bottom column of the table. The reason why the drying speed of the sample D is smaller than that of E is because of the same reason as in the case of the experiment 1 (c).

実験３（凍結乾燥品の製法とセファロチンナトリウム水
溶液の濃度との関係） セファロチンナトリウム水溶液の濃度を変え、以後の条
件は後記実施例１に従って凍結乾燥を行い、得られた凍
結乾燥品の顆粒生成率を調べた。顆粒生成率は篩過法に
より顆粒部分とケーキ部分を分離し、その重量比により
決定した。結果を第３図に示す。第３図は顆粒状晶析を
行うための至適セファロチンナトリウム濃度が２６〜３
６％であることを示している。水溶液濃度が２６％未満
では晶析速度が小さくなり、アモルファス分の生成を避
け得なくなり、また３６％を越えると高粘度なるため無
菌濾過が困難となりかつ、種晶となる結晶の析出が起こ
り易く、部分的に結晶性ケーキを形成し易くなる。Experiment 3 (Relationship between production method of freeze-dried product and concentration of sodium cephalothin solution) Concentration of sodium cephalothin solution was changed, and the subsequent conditions were freeze-dried according to Example 1 to be described later. The production rate was investigated. The granule production rate was determined by separating the granule portion and the cake portion by the sieving method and determining the weight ratio. Results are shown in FIG. Fig. 3 shows that the optimal cephalotin sodium concentration for performing granular crystallization is 26 to 3
It shows that it is 6%. If the concentration of the aqueous solution is less than 26%, the crystallization rate will be low, and the formation of amorphous components will be unavoidable. If it exceeds 36%, aseptic filtration will be difficult due to the high viscosity, and the precipitation of crystals as seed crystals will easily occur. , Partially facilitates the formation of a crystalline cake.

実験４（凍結乾燥品の製法とセファロチンナトリウム水
溶液の凍結前の温度との関係） 濃度２８％のセファロチンナトリウム水溶液を使用し、
凍結直前の液温を０〜４０℃まで変化させた後、冷却速
度０．２℃／分で−４０℃まで冷却、凍結させた後、後
記実施例１に従った凍結乾燥を行い、得られた凍結乾燥
品の顆粒生成率を調べた。その結果を第４図に示す。Experiment 4 (Relationship between production method of freeze-dried product and temperature before freezing of sodium cephalotin aqueous solution) Using an aqueous cefalotin sodium solution having a concentration of 28%,
After changing the liquid temperature immediately before freezing to 0 to 40 ° C., the liquid was cooled to −40 ° C. at a cooling rate of 0.2 ° C./min and frozen, and then freeze-dried according to Example 1 described below to obtain a product. The granulation rate of the freeze-dried product was examined. The results are shown in FIG.

第４図は顆粒状晶析を行うための凍結前の液温の至適条
件が１０〜３０℃であり、より好ましくは１５〜２６℃
であることを示している。凍結前の液温が１０℃未満で
はセファロチンナトリウムの過飽和溶液が擬液晶構造を
形成し、結晶化度の小さい稠密複晶を生じ易くなり、ま
た３０℃を越えると結晶成長が阻害されてアモルファス
分を形成し易くなるので共に好ましくない。FIG. 4 shows that the optimum liquid temperature before freezing for performing granular crystallization is 10 to 30 ° C., more preferably 15 to 26 ° C.
Is shown. If the liquid temperature before freezing is less than 10 ° C, the supersaturated solution of cephalotin sodium forms a pseudo-liquid crystal structure, and a dense double crystal with a low degree of crystallinity is likely to be formed. Both of them are not preferable because they easily form the minute portion.

実験５（凍結乾燥品の製品とセファロチンナトリウム水
溶液の凍結速度との関係） 濃度２８％のセファロチンナトリウム水溶液を使用し、
凍結前の液温を２２℃とした後、冷却速度を変えて−４
０℃まで冷却、凍結した。次いで、後記実施例１に従っ
て凍結乾燥を行い、得られた凍結乾燥品の顆粒生成率を
調べた。結果は第５図に示すように冷却速度が小さい程
顆粒生成率は増大する。この理由は、冷却速度が０．５
℃／分を越えると凍結氷晶およびスケルルトンの厚みが
小さくなり、結晶成長に寄与する溶質の集中度が小さく
なること、および再昇温時の氷晶の形態変化が急激すぎ
て氷晶界面での不均質核生成を阻害することなどによる
ものである。Experiment 5 (Relationship between Freeze-Dried Product and Freezing Rate of Cephalotin Sodium Aqueous Solution) A cephalotin sodium aqueous solution having a concentration of 28% was used,
After setting the liquid temperature before freezing to 22 ° C, change the cooling rate to -4
It was cooled to 0 ° C. and frozen. Then, freeze-drying was performed according to Example 1 described later, and the granule formation rate of the obtained freeze-dried product was examined. As a result, as shown in FIG. 5, the smaller the cooling rate, the higher the granule production rate. The reason for this is that the cooling rate is 0.5
If the temperature exceeds ℃ / min, the thickness of frozen ice crystals and skeletons will be small, the concentration of solutes that contribute to crystal growth will be small, and the morphology change of ice crystals at the time of re-heating will be too rapid to cause ice crystal interface. This is due to the inhibition of heterogeneous nucleation of.

実験６（凍結乾燥品の製法と晶析温度との関係）後記実
施例１に従いセファロチンナトリウム濃度２８％の水溶
液を調製、凍結し、晶析温度を変えて５時間晶析を行っ
た後、さらに後記実施例１に従って凍結乾燥を行い、得
られた凍結乾燥品の顆粒生成率を調べた。結果を第６図
に示す。第６図により顆粒状晶析を行うための晶析温度
条件が−６〜−２℃ときわめてその範囲が狭いことが分
かる。晶析温度が−６℃以下では結晶化度の低い稠密複
晶およびアモルファス分から成る結晶性凍結乾燥ケーキ
が得られ、−２℃以上では氷晶が完全融解するためアモ
ルファス分が形成される。以下に実施例および参考例を
挙げ、本発明方法を具体的に説明する。Experiment 6 (Relationship between production method of lyophilized product and crystallization temperature) An aqueous solution having a cefalotin sodium concentration of 28% was prepared and frozen according to Example 1 to be described later, and crystallization was performed for 5 hours while changing the crystallization temperature. Further, freeze-drying was performed according to Example 1 described below, and the granule formation rate of the obtained freeze-dried product was examined. Results are shown in FIG. It can be seen from FIG. 6 that the crystallization temperature condition for performing granular crystallization is -6 to -2 ° C, which is a very narrow range. When the crystallization temperature is −6 ° C. or lower, a crystalline freeze-dried cake composed of a dense double crystal having a low degree of crystallinity and an amorphous component is obtained, and at −2 ° C. or higher, the ice crystal is completely melted to form an amorphous component. Hereinafter, the method of the present invention will be specifically described with reference to Examples and Reference Examples.

実施例１ セファロチンナトリウム粉末２８０ｇを局方蒸留水７２
０ｇに５０℃で加熱溶解し、直ちに５℃に急冷する。次
いで２０℃に昇温し、１時間した後０．２μのメンブラ
ンフィルターで濾過し、容量１４mlのバイアルに３．３
mlずつ分注する。このバイアルを冷却速度０．５℃／分
で−４０℃まで冷却し、次いで−４℃に昇温し、この温
度に３時間保持して晶析させる。次いで−４０℃に再冷
却した後、棚温３０℃、真空度０．４mbarで５時間、次
いで５０℃、０．０３mbarで１．５時間、凍結真空乾燥
機で乾燥させる。この操作で結晶化度が１００％である
粒径０．３mmの顆粒状の結晶性凍結乾燥品が得られる。Example 1 280 g of sodium cephalothin powder was mixed with 72 g of pharmacopoeia distilled water.
It is dissolved in 0 g by heating at 50 ° C and immediately cooled to 5 ° C. Then, the temperature was raised to 20 ° C., and after 1 hour, the mixture was filtered through a 0.2 μ membrane filter and then 3.3 ml in a vial having a volume of 14 ml.
Dispense ml by ml. The vial is cooled to −40 ° C. at a cooling rate of 0.5 ° C./min, then heated to −4 ° C. and kept at this temperature for 3 hours for crystallization. After recooling to −40 ° C., the product is dried in a freeze-vacuum dryer at a shelf temperature of 30 ° C. and a vacuum degree of 0.4 mbar for 5 hours, and then at 50 ° C. and 0.03 mbar for 1.5 hours. By this operation, a granular crystalline freeze-dried product having a crystallinity of 100% and a particle size of 0.3 mm is obtained.

実施例２ セファロチンナトリウム粉末３００ｇを局方蒸留水７０
０ｇに６５℃で加熱溶解し、直ちに５℃に急冷する。次
いで１８℃に昇温し、１時間保持した後、０．２μのメ
ンブランフィルターで濾過し、容量２０mlのバイアルに
６．２mlずつ分注する。このバイアルを冷却速度０．２
℃／分で−４０℃まで冷却し、次いで−３．５℃に昇温
し、この温度に４時間保持して晶析させる。次いで−４
０℃に再冷却した後、棚温３０℃、真空度０．２mbarで
１０時間、次いで５０℃、０．０３mbarで１．５時間、
凍結真空乾燥機で乾燥させる。この操作で結晶化度が１
００％である粒径０．８mmの顆粒状の結晶性凍結乾燥品
が得られる。Example 2 300 g of sodium cephalothin powder was added to 70 g of pharmacopoeia distilled water.
Heat and dissolve in 0 g at 65 ° C and immediately quench to 5 ° C. Then, the temperature is raised to 18 ° C., the temperature is maintained for 1 hour, the mixture is filtered through a 0.2 μ membrane filter, and 6.2 ml is dispensed into a vial having a volume of 20 ml. Cool this vial to a cooling rate of 0.2
The mixture is cooled to -40 ° C at a rate of ° C / min, then heated to -3.5 ° C, and kept at this temperature for 4 hours for crystallization. Then -4
After recooling to 0 ° C., shelf temperature 30 ° C., vacuum degree 0.2 mbar for 10 hours, then 50 ° C. 0.03 mbar for 1.5 hours,
Dry in a freeze vacuum dryer. Crystallinity is 1 by this operation
A granular crystalline lyophilized product with a particle size of 0.8 mm is obtained, which is 00%.

実施例３ セファロチンナトリウム粉末３２０ｇを局方蒸留水６８
０ｇに７０℃で加熱溶解し、直ちに５℃に急冷する。次
いで２５℃に昇温し、１．５時間保持した後、０．２μ
のメンブランフィルターで濾過し容量１００mmのバイア
ルに６．０mlずつ分注する。このバイアルを冷却速度
０．１℃／分で−３５℃まで冷却し、次いで−４．５℃
に昇温し、この温度に４時間保持して晶析させる。次い
で−４０℃に再冷却した後、棚温２５℃、真空度０．３
mbarで４時間、次いで５０℃、０．０３mbarで１時間、
凍結真空乾燥機で乾燥させる。この操作で結晶化度１０
０％である粒径０．２mmの顆粒状の結晶性凍結乾燥品が
得られる。Example 3 320 g of sodium cephalothin powder was mixed with 68 g of pharmacopoeia distilled water.
It is dissolved in 0 g by heating at 70 ° C. and immediately cooled to 5 ° C. Then, after raising the temperature to 25 ° C. and holding for 1.5 hours, 0.2 μ
Filter with a membrane filter of No. 1 and dispense 6.0 ml into a vial having a volume of 100 mm. The vial was cooled to -35 ° C at a cooling rate of 0.1 ° C / min, then -4.5 ° C.
The temperature is raised to and maintained at this temperature for 4 hours for crystallization. Then, after recooling to -40 ° C, shelf temperature 25 ° C, vacuum degree 0.3.
mbar for 4 hours, then 50 ° C., 0.03 mbar for 1 hour,
Dry in a freeze vacuum dryer. Crystallinity 10 by this operation
A granular crystalline lyophilized product with a particle size of 0.2 mm, which is 0%, is obtained.

参考例１ セファロチンナトリウム粉末１１４ｇを局方蒸留水２０
０mlに溶解し、更に蒸留水を加えて全量５００mlとし、
その５mlずつを容量１４mlのバイアルに分注する。次い
でバイアルを−２５℃の棚に置いて２時間冷して凍結
し、次いで−７℃に昇温し、この温度に１５時間保持し
て晶析させる。ついで棚温１０℃、真空度０．１４mbar
で２０時間、次いで棚温３５℃、０．０３mbarで１時
間、凍結真空乾燥機により乾燥する。表面の結晶化度が
９４．５％である結晶性白色ケーキが得られる。Reference Example 1 114 g of sodium cephalothin powder was mixed with 20 g of pharmacopoeia
Dissolve in 0 ml and add distilled water to make a total volume of 500 ml.
Dispense 5 ml each into a vial having a volume of 14 ml. The vials are then placed on a shelf at -25 ° C to cool for 2 hours to freeze, then warmed to -7 ° C and held at this temperature for 15 hours for crystallization. Then the shelf temperature is 10 ° C and the vacuum degree is 0.14 mbar.
For 20 hours and then at a shelf temperature of 35 ° C. for 1 hour at 0.03 mbar in a freeze vacuum dryer. A crystalline white cake with a surface crystallinity of 94.5% is obtained.

参考例２ セファロチンナトリウム粉末２８０ｇを局方蒸留水７２
０ｇに５０℃で加熱溶解し、直ちに５℃に急冷する。次
いで、セファロチンナトリウムの種晶（約１×０．３
μ）の５０％エタノールスラリー１０μｌ（水溶液の５
×１０^-3％）をエッペンドルフタイプピベツトを用いて
滴下し、マグネチックスタラーで約５分間攪拌した後２
０μリジメッシュフィルターで濾過し、容量１４mlのバ
イアルに３．３mlずつ分注する。このバイアルを−４０
℃に再冷却した後、棚温３０℃真空度０．４mbarで８時
間、次いで５０℃、０．０３mbarで１．５時間凍結真空
乾燥機で乾燥させる。この操作で結晶化度が１００％で
ある結晶性白色ケーキが得られる。Reference Example 2 280 g of cephalothin sodium powder was added to 72 g of pharmacopoeia distilled water.
It is dissolved in 0 g by heating at 50 ° C and immediately cooled to 5 ° C. Next, seed crystals of cephalotin sodium (about 1 × 0.3
μ) 50% ethanol slurry 10 μl (5% of aqueous solution)
(× 10 ^-3 %) was added dropwise using an Eppendorf type pipette, and the mixture was stirred with a magnetic stirrer for about 5 minutes and then 2
Filter through a 0μ Rigimesh filter and dispense into 3.3 ml aliquots in 14 ml vials. -40 this vial
After re-cooling to 0 ° C., it is dried in a freeze-vacuum dryer at a shelf temperature of 30 ° C. and a vacuum degree of 0.4 mbar for 8 hours and then at 50 ° C. and 0.03 mbar for 1.5 hours. By this operation, a crystalline white cake having a crystallinity of 100% is obtained.

【図面の簡単な説明】[Brief description of drawings]

第１図は本発明方法および他の公知方法によって得られ
たセファロチンナトリウムの凍結乾燥品の経時的着色度
の度合を示すグラフ、第２図は第１図の場合と同じ試料
についてのセファロチンナトリウムの経時的な含量変化
を示すグラフ、第３図は凍結乾燥に付すセファロチンナ
トリウム水溶液の濃度とその顆粒生成率との関係を示す
グラフ、第４図は凍結前のセファロチンナトリウム水溶
液の温度とその顆粒生成率との関係を示すグラフ、第５
図はセファロチンナトリウム水溶液の凍結までの冷却速
度とその顆粒生成率との関係を示すグラフ、第６図は晶
析温度とセファロチンナトリウムの顆粒生成率との関係
を示すグラフである。FIG. 1 is a graph showing the degree of coloring over time of a freeze-dried product of cephalotin sodium obtained by the method of the present invention and other known methods, and FIG. 2 is the same sample as in the case of FIG. Fig. 3 is a graph showing the change in sodium content over time, Fig. 3 is a graph showing the relationship between the concentration of the cefarotin sodium aqueous solution subjected to freeze-drying and its granule formation rate, and Fig. 4 is the temperature of the cefarotin sodium aqueous solution before freezing. Graph showing the relationship between the rate of granule formation and
FIG. 6 is a graph showing the relationship between the cooling rate of an aqueous cephalotin sodium solution until freezing and the granule formation rate thereof, and FIG. 6 is a graph showing the relationship between the crystallization temperature and the granule formation rate of cephalothin sodium.

───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭51−123813（ＪＰ，Ａ) 特開 昭56−120615（ＪＰ，Ａ) 特開 昭60−11491（ＪＰ，Ａ) ─────────────────────────────────────────────────── ─── Continuation of front page (56) Reference JP-A-51-123813 (JP, A) JP-A-56-120615 (JP, A) JP-A-60-11491 (JP, A)

Claims (2)

【特許請求の範囲】[Claims] 【請求項１】水溶液の濃度が２６〜３６重量％であり、
かつ、その温度が１０〜３０℃であるセファロチンナト
リウム過飽和水溶液を、０．５℃／分以下の冷却速度で
−１５℃以下に徐冷凍結後、−５〜−２℃まで昇温せし
めて晶析を行い、次いで−３０〜−４０℃に再冷却後凍
結乾燥工程に付すことを特徴とするセファロチンナトリ
ウムの結晶性凍結乾燥製剤の製造法。1. The concentration of the aqueous solution is 26 to 36% by weight,
Also, the supersaturated aqueous solution of sodium cephalotin having a temperature of 10 to 30 ° C is slowly cooled to -15 ° C or less at a cooling rate of 0.5 ° C / min or less, and then heated to -5 to -2 ° C. A method for producing a crystalline freeze-dried preparation of sodium cephalotin, which comprises subjecting to crystallization, recooling to -30 to -40 ° C, and then subjecting to a freeze-drying step. 【請求項２】セファロチンナトリウム凍結水溶液の晶析
温度が−４．５〜−３．５℃である特許請求の範囲第
（１）項記載の製造法。2. The method according to claim 1, wherein the frozen aqueous solution of sodium cephalotin has a crystallization temperature of -4.5 to -3.5 ° C.