Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai
Reexamination Certificate
1999-03-08
2002-05-07
Low, Christopher S. F. (Department: 1653)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Peptide containing doai
C514S866000, C530S303000, C530S308000, C530S324000
Reexamination Certificate
active
06384016
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to stable, injectable aqueous solutions of peptides such as glucagon, glucagon-like peptide-1 (GLP-1, e.g. GLP-1(7-37) and GLP-1(7-36) amide) and analogues and derivatives thereof.
BACKGROUND OF THE INVENTION
Glucagon and glucagon-like peptide 1 (GLP-1) are polypeptide hormones both derived from proglucagon. Upon tissue-specific processing, glucagon is produced in the pancreas, while GLP-1 is predominantly secreted in the intestine. Physiologically, both peptides play major roles in the regulation of blood glucose. Glucagon is directly involved via glycogenolytic as well as gluconeogenetic effects. GLP-1 is indirectly involved via stimulation of insulin and inhibition of glucagon secretion. Thus, the two peptides affect the blood glucose in opposite directions.
Due to its glycogenolytic effect on the liver, glucagon is used for the treatment of acute hypoglycemia (e.g. “insulin shock”). In addition, glucagon excerts a spasmolytic effect on smooth muscles, and this effect is used clinically in connection with several imaging procedures, especially radiology.
GLP-1 fragments such as GLP-1 (7-37), GLP-1 (7-36) amide and functional analogues thereof (hereinafter designated GLP-1 compounds) stimulate pancreatic insulin release in the presence of glucose, and this finding suggests that GLP-1 compounds may be useful in the treatment of type II diabetes.
Glucagon (29 amino acids; see e.g.
The Merck Index
, 10
th
Edition for the amino acid sequence) and GLP-1 compounds (typically about 29-31 amino acids) are single chain highly homologous peptides. While little information is available on the solution behaviour of GLP-1 compounds, the physico-chemical properties of glucagon are well described in the literature (J.Biol.Chem. 244, 6675-6679 (1969); Nature 257, 751-757 (1975); FEBS Letts. 119, 265-270 (1980)). The peptide exhibits very low solubility in water at a pH in the range of about 4-9. Outside this region, aqueous solutions are generally metastable and prone to gel formation or flocculation within hours of their preparation.
Accordingly, glucagon compositions for injection are currently marketed in the form of a lyophilized powder which is to be reconstituted using a suitable diluent shortly before use. The resulting acidic solution may then be injected. In contrast with the single-dose compositions of glucagon used for acute hypoglycemia and the motility indication in radiology, the utility of GLP-1 compounds for the treatment of type II diabetes generally involves multiple-dose soluble compositions. Given the instability of glucagon, the formulation of such compositions is inherently difficult.
Thus, there is a need for stable aqueous formulations of peptides such as glucagon and GLP-1 compounds. In the case of glucagon, the availability of a ready-to-use composition is desirable for emergency treatment of acute hypoglycemia, enabling the patient or another person present to treat the hypoglycemic incidence immediately. Also for the mobility indication for glucagon used in radiology, and for the type II diabetes indication for GLP-1 compounds, the availability of a ready-to-use aqueous composition provides obvious advantages in terms of convenience and safety.
Glucagon compositions solubilized by the addition of detergent have received relatively little attention in the literature. British patent No. 1 202 607 describes the use of either cationic (quarternary ammonium bases) or anionic (alkyl-aryl-sulphonates), monovalent detergents. These compounds provide enhanced solubilization of 1 mg/ml glucagon over a limited pH range using a 6-fold molar excess of detergent.
A series of other studies on glucagon-detergent interactions employ much lower glucagon concentrations and do not address the effect of detergent on peptide solubility. Thus, in studies using 0-200 &mgr;M of the amphoteric (twitterionic) detergent hen egg lysolecithin, binding of detergent was shown to induce partial helical structure in solutions of glucagon containing about 0.02 mg/ml peptide (J.Biol.Chem. 247, 4986-4991; 4992-4995; (1972)). Similarly, in studies employing dilute (0.005-0.05 mg/ml) solutions of glucagon, lysolecithin was found more efficient than the anionic detergents dipalmitoyl phosphatidic acid and sodium dodecyl sulfate (SDS) in inducing partial helical structure in glucagon (Biopolymers 21, 1217-1228 (1982); Biopolymers 22, 1003-1021 (1983)).
At the other extreme, a few examples are available characterizing the interaction between glucagon and concentrated detergent solutions. These include the binding of dilute glucagon to micelles of the anionic detergent SDS at 25 mM, pH 2 (Biochemistry 19, 2117-2122, (1980)), and the solubilization of glucagon (up to 20 mg/ml) via binding to micelles of the amphoteric dodecylphosphocholine present in a 40-fold molar excess (e.g. 3.5 mg/ml glucagon is solubilized using 40 mM detergent at neutral pH; Biochim.Biophys.Acta 603, 298-312, (1980))
With the exception of a single example describing the solubilization of 20 mg/ml GLP-1 (7-36) amide bound to micelles of dodecylphosphocholine at 280 mM (cf. the glucagon case above, Biochemistry 33, 3532-3539, 1994), no reports are available on the binding of detergents by GLP-1 compounds and the resulting effect on peptide solubility and stability. In contrast, several ways of preparing prolonged delivery of GLP-1 compounds have been described (e.g. European patent application No. 0619322 and column 6 of U.S. Pat. No. 5,120,712). The possibilities include use of polymers, incorporation into particles of polymer material or into an oil suspension, addition of zinc ion, protamine sulphate and/or phenolic compounds, and preparation of amorphous/crystalline formulations.
BRIEF DESCRIPTION OF THE INVENTION
It has now surprisingly been found that soluble and stable formulations of glucagon and GLP-1 compounds may be prepared by means of addition of detergents carrying multiple charges (i.e. two or more negative, two or more positive, or both positive and negative charges). When added in an amount corresponding to e.g. about 0.5-20 moles of detergent per mole of peptide, this class of detergents is capable of solubilizing pharmaceutically relevant concentrations of glucagon and GLP-1 compounds in the entire pH range from 4 to 9. The expanded pH range allows optimal selection of conditions to suppress chemical degradation of the solubilized peptide. Furthermore, preferred detergents consist of naturally occurring building blocks which are more biologically acceptable than those previously described.
Thus, in its broadest aspect, the present invention relates to a pharmaceutical composition in the form of a solution comprising a polypeptide containing an amphiphatic &agr;-helix which is stabilized by interaction with a detergent carrying multiple charges. Such polypeptides include, but are not limited to, glucagon, GLP-1 compounds, corticotropin (ACTH), calcitonin, and functional analogues thereof.
One aspect of the invention relates to an aqueous composition in the form of a solution comprising at least one peptide selected from glucagon, GLP-1, and analogues and derivatives thereof together with a stabilizing and solubilizing amount of at least one detergent, said detergent having at least 2 positive charges, at least 2 negative charges, or a combination of at least one positive charge and at least one negative charge, the peptide or analogue or derivative thereof being present in a concentration of at least about 0.1 mg/ml, with the proviso that the detergent is not dodecyl phosphocholine.
Another aspect of the invention relates to a method for stabilizing an aqueous composition in the form of a solution comprising at least one peptide selected from glucagon, GLP-1 and analogues and derivatives thereof by adding to the composition a stabilizing and solubilizing amount of at least one such detergent having multiple charges.
In a further embodiment, the invention relates to a method for treating insulin dependent or non-insulin dependent diabetes mellitus or obesity in a
Green, Esq. Reza
Gregg, Esq. Valeta A.
Low Christopher S. F.
Mohamed Abdel A.
Novo Nordisk A S
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