title: The improved dissolution and prevention of ampoule breakage attained by the introduction of pretreatment into the production process of the lyophilized formulation of recombinant human Interleukin-11 (rhIL-11).
abstract: Lyophilized protein formulations sometimes pose problems such as the formation of a cloudy solution upon reconstitution. Ampoule or vial breakage can also occur during the production processes of lyophilized pharmaceutical products. Various efforts have been made to overcome those difficult problems. In this study, we introduce a particular temperature program into the production process of a recombinant human Interleukin-11 (rhIL-11) lyophilized formulation containing sodium phosphates (Na2HPO4/NaH2PO4, pH 7.0) and glycine in an attempt to improve its dissolution properties and to prevent ampoule breakage from occurring. The formulation was pretreated by nucleating ice and maintaining the solution overnight at a temperature of -6 degrees C. The solution was then completely frozen at a lower temperature. This pretreatment proved successful in not only producing a lyophilized cake which readily disintegrated and dissolved in the reconstitution media, but also prevented ampoule breakage from occurring during the production processes. In contrast, a lyophilized cake produced without the pretreatment created a cloudy solution particularly when reconstituted using water for injection contaminated with aluminum (Al3+), although the solution became transparent within 20-30 min. The pretreatment induced the crystallization of sodium dibasic phosphate (Na2HPO4) in the freeze-concentrate whereas direct freezing without the pretreatment did not crystallize the salt. Thermal analyses (DSC and TMA) showed that amorphous sodium dibasic phosphate in the freeze-concentrate became crystallized upon heating, accompanied by an increase in volume, which probably caused the ampoule breakage that occurred without the pretreatment. Although power X-ray diffraction (PXRD) experiments suggested that, with or without the pretreatment, glycine assumed the beta-form and sodium phosphate stayed amorphous in the final products, an electrostatic interaction between dibasic phosphate anions and rhIL-11, a highly cationic protein, would only exist in the lyophilized cake produced without the pretreatment. This interaction is highly likely because aluminum facilitates the formation of a cloudy solution upon reconstitution possibly by using the divalent anions which effectively reduce electrostatic repulsions between aluminum and the protein to form an aggregate structure that is not readily soluble. The pretreatment would circumvent the interaction by crystallizing the sodium salt before freezing creating a relatively soluble lyophilized cake that is much less sensitive to aluminum.
authors.affiliation: Novel Pharmaceutical Laboratories, Yamanouchi Pharmaceutical Co. Ltd., 180 Ozumi, Yaizu, Shizuoka 425-0072, Japan. hirakura.yutaka@yamanouchi.co.jp
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hirakura
authors.forename: Seiki
authors.initials: S
authors.lastname: Kojima
authors.forename: Akira
authors.initials: A
authors.lastname: Okada
authors.forename: Shigeharu
authors.initials: S
authors.lastname: Yokohama
authors.forename: Shoji
authors.initials: S
authors.lastname: Yokota
mesh_terms: D002626:Chemistry, Pharmaceutical / Q000379:methods* / Q000592:standards; D003460:Crystallization; D004055:Differential Thermal Analysis / Q000379:methods; D004304:Dosage Forms / Q000592:standards; D004349:Drug Packaging / Q000379:methods* / Q000592:standards*; D004355:Drug Stability; D005612:Freeze Drying / Q000295:instrumentation / Q000379:methods*; D005898:Glass; D005998:Glycine / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007267:Injections; D017370:Interleukin-11 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007564:Japan; D010710:Phosphates / Q000737:chemistry; D011409:Propylene Glycols / Q000737:chemistry; D012995:Solubility; D013499:Surface Properties; D013501:Surface-Active Agents / Q000737:chemistry; D013696:Temperature; D022722:Thermogenesis; D013997:Time Factors; D014961:X-Ray Diffraction / Q000379:methods
chemical_list: D004304:Dosage Forms; D017370:Interleukin-11; D010710:Phosphates; D011409:Propylene Glycols; D013501:Surface-Active Agents; C041787:1,3-propanediol; C018279:sodium phosphate; D005998:Glycine