Insulin lyophilized

The mechanisms and kinetics of degradation of human insulin lyophilized powders at pH 2-5 were studied as a function of water content.. .. The degradation rate inaeased with decreasing apparent pH ( pH ), yielding, at any given water content, solid state pH rate profiles parallel to the solution pH rate profile. The pH dependence was accounted for in terms of the fraction of insulin A21 carboxyl in its neutral form, with an apparent of - 4, independent of water content."... 

Lyophilized copolymer was ground down to colloidal dimensions (<1 iJim) using a laboratory planetary mill (Pulverisette, Fritsch GmbH). A 110-mg sample of copolymer powder, 20 mg of bovine insulin, and 20 mg of GOD were mixed, and the mixture was compressed into a disk-shaped matrix of 5-mm thickness and 15-mm diameter. 

Strickley, R.G., and B.D. Anderson, Solid-state stability of human insulin. I. Mechanism and the effect of water on the kinetics of degradation in lyophiles from pH 2-5 solutions. Pharm Res, 1996.13(8) 1142-53. 

Insulin Trehalose (0.5% w/v) Lyophilized -2 fold increased stability at 35°C [25]... 

Protein-based drugs have been formulated mainly as stable liquids or in cases where liquid stability is limiting as lyophilized dosage forms to be reconstituted with a suitable diluent prior to injection. This is because their delivery has been limited primarily to the parenteral routes of intravenous (IV), subcutaneous (SC), or intramuscular (IM) administration. There are a few drugs that have been developed for pulmonary delivery, such as rhDNase (Pulmozyme ) and an inhalable formulation of insulin (e.g., Exubra ). However, even such drugs have been formulated as either liquid or lyophilized or spray-dried powders. This chapter will focus only on excipients that are applicable to liquid and lyophilized protein formulations. 

Costantino HR, Langer R, Klibanov AM. Moisture induced aggregation of lyophilized insulin. Pharm Res 1994 11 21-29. 

Ten severely hyperglycemic patients with type 2 diabetes taking oral agents were treated at random with two injections of isophane daily or with lyophilized nasal insulin before each meal, with an added injection of subcutaneous isophane when necessary (261). The periods were separated by 2 months. Nasal insulin produced control of diabetes comparable to isophane, except in three patients. Adverse effects included transient pruritus, sneezing and rhinorrhea, and chronic nasal crusts. One patient was withdrawn because of cough and dizziness after each nasal dose. 

Parenteral 1 mg lyophilized powder to reconstitute for injection 1 See Table 41-4 for Insulin Preparations. 

A solution of insulin is added to an aqueous suspension of Technosphere particles. The insulin is adsorbed to the surface of the particles to form TI. The solvent is then removed from the suspension by lyophilization to produce the bulk drug powder. TI has a mean geometric diameter of 2.0 p,m and an aerodynamic diameter of 2.5 xm. 

I-Tyr-A 14-insulin (e.g. human, porcine or bovine insulin) is prepared according to routine procedures. Aliquots in buffer solution (0.05 M phosphate buffer, pH 7.4,1 % BSA, 0.05 % NaN3) are lyophilized. 

I-insulin tracer lyophilized, is dissolved in 20 ml H20 bidest. It can be stored up to 24 hours at 2-8 °C after reconstitution. 

Crystalline cellulose, hydroxypropyl cellulose, and Carbopol 934 have been studied in combination with lyophilized insulin as bioadhesive powder dosage forms for nasal delivery. Each formulation tested resulted in an decrease in plasma glucose level after nasal administration in dog and rabbit models. The most effective formulation, crystalline cellulose blended with insulin, decreased the plasma glucose level to 49% of the control value. In ternary systems the lyophilized Carbopol 934 and insulin blend with crystalline cellulose powder has been the most effective, leading to a hypoglycemia on the order of one-third of the effect obtained after intravenous injection of the same dose of insulin. The plasma glucose levels obtained in the volunteers after administration of the insulin-Carbopol-crystalline cellulose powder formulation were quite variable [38],... 

Powder Lyophilization Insulin Starch-Carbopol 974P, maltodextrin-Carbopol 974P In rabbits The highest absolute bioavailability obtained was 14.4% 46... 

Grasbeck in 1955 (G26, G27) collected normal human gastric juice on ice, after i.v. stimulation with insulin, neutralized it in vitro, removed mucus by centrifugation, and dialyzed and lyophilized the supernatant. 

Proteins were lyophilized directly in the reaction vessels. Only in the case of insulin at LpH 7.5 was it necessary to lyophilize the protein from a large volume and transfer it to the reaction vessel. Insulin (20 mg) was lyophilized from a solution of 1 mM sodium phosphate buffer (40 ml), pH 7.5 and 40 mM sodium metaborate buffer (1 ml), pH 10. Ribonuclease and a-chymotrypsin were lyophilized from a solution of 40 mM sodium phosphate buffer (1 ml), pH 7.5 and 40 mM sodium metaborate buffer (1 mi), pH 10. Anhydrous octane (2 ml) was added to the protein and the medium was sonicated until the protein was finely dispersed, at which time [ Cjiodomethane (100 gl) was added. To prevent the loss of... 

There is a wide range of devices available to suit different applications. Some are ideally suited to chronic injections of varying doses of insulin several times a day, others are more suited to weekly dosing of the same dose of a therapeutic protein or even a monoclonal antibody. Still others will be capable of conveniently reconstituting lyophilized drug formulations... 

Those fractions containing insulin-like material were pooled and the ethanol was removed by gel filtration on Sephadex G-25 disposable columns (Pharmacia PD-10) (j 7). The eluates were pooled and lyophilized. The material was then gel filtered on a Sephadex G-50 (fine) column and a peak of insulin-like immunoactivity was recovered in the region where mammalian insulins migrate on Sephadex G-50 (Figure 4). 

Those fractions containing the immunoactive insulin were pooled, lyophilized and tested for insulin bioactivity in a bioassay which measures the incorporation of 3H-glucose into toluene soluble lipids using adipocytes from male rats. (J 3). The amount of insulin bioactivity recovered from the extract was similar to the amount of immunoactivity obtained in the insulin radioimmunoassay. In addition the bioactivity was almost completely removed in the presence of an excess of anti-insulin antibody as well as by an antibody which blocks the insulin receptor (data not shown) (Figure 5) (4 ). 

Sample preparation Dissolve 70 mg trinitrobenzenesulfonic acid in 1 mL 100 mM pH 8.2 sodium bicarbonate and immediately add an aliquot to 50 volumes of 10 mg/mL insulin in 100 mM pH 8.2 sodium bicarbonate, let stand in the dark at room temperature for 2 h, add to a 150 X 60 column of Sephadex G25 made up in 100 mM pH 8.2 sodium bicarbonate, collect the major colored band and lyophilize it. Reconstitute, inject an aliquot. 

Flores-Ferntodez, G.M., Sold, R.J., Griebenow, K. (2009) The relation between moisture-induced aggregation and structural changes in lyophilized insulin. J Pharm Pharmacol, 61 (11), 1555-1561. 

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