JPH06321800A - Lyophilized preparation - Google Patents

Abstract

(57) Summary [Objective] The present invention provides a lyophilized preparation of a peptide or protein. [Structure] To provide a freeze-dried preparation of a peptide or protein characterized by containing gluconate.

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a lyophilized preparation of peptide or protein containing gluconate. [0002] Peptides and proteins are unstable in a solution state, and operations such as lyophilization are required to provide stable peptide or protein preparations. However, it is known that upon freeze-drying, peptides and proteins are inactivated or insolubilized due to decomposition and denaturation due to various factors. When freeze-drying, a conventionally known stabilizing base for saccharides such as mannitol, maltose, glucose and lactose, and hydrophilic amino acids such as glycine and arginine is used. However, some of the peptides or proteins do not give satisfactory results,
The present inventors have further earnestly studied the stabilizing base. The present invention is characterized in that in the preparation of a freeze-dried preparation of a peptide or protein, gluconate is added as a stabilizing base to provide a freeze-dried preparation that is stable for a long period of time. Have. Examples of the gluconate used in the present invention include calcium gluconate, magnesium gluconate, sodium gluconate, potassium gluconate, and the like. Particularly preferred is magnesium gluconate. Gluconate alone or 2
It can be used as a mixture of one or more kinds, and may be used in combination with other stabilizing bases which have been conventionally used.
The addition amount of the gluconate used in the present invention is 1/10 to 100,000 parts by weight to 1 part by weight of the peptide or protein. The concentration is preferably 0.5 to 10%. The pH of the solution for preparing the freeze-dried preparation of the present invention is preferably 4.0 to 7.5, and in some cases, a pH adjuster, physiologically acceptable additives, diluents, excipients, etc. may be added. Can be mixed. The freeze-dried preparation of the present invention can be prepared by freezing a gluconate-containing peptide or protein solution at -15 ° C to -60 ° C and then drying under reduced pressure. The peptide or protein in the present invention is, in particular, a physiologically active peptide or protein, and examples thereof include tuftsin, vasopressin, luteinizing hormone-releasing hormone, human growth hormone, growth hormone-releasing factor,
Substance-P, insulin, tetragastrin, basic fibroblast growth factor, acidic fibroblast growth factor, hepatocyte growth factor, epidermal growth factor, bone morphogenetic factor, colony stimulating factor, insulin-like growth factor, nerve growth factor , Stem cell growth factor, tumor necrosis factor, vascular endothelial cell growth factor,
Examples include various interleukins. Example 1 Water for injection containing 2.5% magnesium gluconate,
Tuftsin, vasopressin, luteinizing hormone-releasing hormone, substance P, insulin, epidermal growth factor or basic fibroblast growth factor 50μ each
g / ml was added, and 1 ml was filled in 1 vial, kept at -40 ° C for 2 hours, frozen, and then dried under reduced pressure to prepare a freeze-dried preparation. Example 2 Tuftsin, vasopressin, luteinizing hormone-releasing hormone, substance P, insulin, epidermal growth factor or basic fibroblast growth factor (50 μg) were added to water for injection containing 3.0% potassium gluconate. /
Add 1 ml to 1 vial, hold at -20 ° C for 2 hours, freeze, and dry under reduced pressure.
A lyophilized formulation was prepared. Example 3 Tuftsin, vasopressin, luteinizing hormone-releasing hormone, substance P, insulin, epidermal growth factor or basic fibroblast growth factor (50 μg) were added to water for injection containing 2.0% sodium gluconate.
/ Ml, 1 ml was filled in 1 vial, and the mixture was kept at -50 ° C for 1 hour, frozen, and then dried under reduced pressure.
A lyophilized formulation was prepared. Test Example 1 The freeze-dried preparation prepared in Example 1 was stored under severe conditions of 60 ° C. for 2 weeks, and the stability of bioactive peptides or proteins was compared. As a control, instead of 2.5% magnesium gluconate in Example 1, 2.
5% glycine or 2.5% mannitol was used as a comparative control formulation. The results are shown in Table 1. Table 1 Reagent Base Remaining Rate (2 weeks) Tuftsin Glycine 0 Mannitol 0 Magnesium Gluconate 101.0 Vasopressin Glycine 8.1 Mannitol 43.2 Magnesium Gluconate 60.4 Luteinizing Glycine 63.6 Release Hormone Mannitol 75.8 Magnesium Gluconate 98.2 Substance-P Glycine 45.3 Mannitol 32.8 Magnesium Gluconate 63.6 Insulin Glycine 25.7 Mannitol 14.5 Magnesium Gluconate 53.2 Epidermal Growth Factor Glycine 0 Mannitol 0 Glucon Magnesium acid 63.2 Basic fibroblast Glycine 5.5 Growth factor mannitol 0 Magnesium gluconate 73.2 [Effect of the invention] These tests Et al., Freeze-dried preparation of the present invention showed excellent stability.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Office reference number FI technical display location A61K 37/34 8314-4C 47/26 J 7433-4C

Claims (1)

[Claims] 1. A freeze-dried preparation of a peptide or protein, which contains a gluconate.