Formulations for IL-11

Drug – bio-affecting and body treating compositions – Lymphokine – Interleukin

Reexamination Certificate

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C530S351000

Reexamination Certificate

active

06270757

ABSTRACT:

FIELD OF INVENTION
The present invention relates to novel formulations comprising IL-11
BACKGROUND OF THE INVENTION
A variety of regulatory molecules, known as cytokines, have been identified including interleukin- 11 (IL-11). IL-11 stimulates a variety of hematopoietic and immune functions. The various protein forms of IL-11 and DNA encoding various forms of IL-11 activity are described in Bennett, et al., U.S. Pat. No. 5,215,895 (June 1, 1993); McCoy, et al., U.S. Pat. No. 5,270,181 (Dec. 14, 1993); and McCoy, et al., U.S. Pat. No. 5,292,646 (Mar. 8, 1994), and incorporated herein by reference. Thus, the term “IL-11” includes protein produced by recombinant genetic engineering techniques; purified from cell sources producing the factor naturally or upon induction with other factors; or synthesized by chemical techniques; or a combination of the foregoing.
To maximize the pharmacological benefit of any protein, it is essential to have finished dosage forms that are stable, easily and reproducibly manufactured, and designed for standard routes of administration. Specifically, it is desirable to have stable, concentrated forms of bulk protein, e.g., IL-11 which, in turn, are suitable for further manufacture of finished dosage forms of protein, which can then be administered e.g., via sub cutaneous injection.
In both bulk protein and finished dosage forms, protein stability can be affected by such factors as ionic strength, pH, temperature, repeated cycles of freeze/thaw and shear forces. Active protein may be lost as a result of physical instabilities, including denaturation and aggregation (both soluble and insoluble aggregate formation), as well as chemical instabilities, including, for example, hydrolysis, deamidation, and oxidation, to name just a few. For a general review of stability of protein pharmaceuticals, see, for example, Manning, et al., Pharmaceutical Research 6:903-918 (1989). In addition, it is desirable to maintain stability in the absence of carrier proteins.
While it is widely appreciated that these possible protein instabilities can occur, until a protein has been studied it is impossible to predict the particular instability problems that a particular protein may have. Any of these instabilities can potentially result in the formation of a protein or protein by-product or derivative having lowered activity, increased toxicity, and/or increased immunogenicity. Indeed, protein precipitation can lead to thrombosis, non-homogeneity of dosage form and immune reactions. Thus, the safety and efficacy of any pharmaceutical formulation of a protein is directly related to its stability.
Accordingly, there continues to exist a need in the art for methods for improving protein stability during the concentration process as well as providing stability in the absence of other carrier proteins in a concentration sufficiently high for various routes of administration including, e.g., sub cutaneous injection, intra venous injection.
BRIEF SUMMARY OF THE INVENTION
One aspect of the present invention provides novel compositions and methods for obtaining concentrated preparations of IL-11, useful as bulk drug product.
Another aspect of the present invention provides compositions comprising formulations of IL-11 of a concentration, useful for administration in final dosage forms.
Preferred compositions include IL-11 and glycine, and optionally include a buffering agent. Preferred glycine concentrations range from 100 mM to 300 mM, with 300 mM most preferred. IL-11 concentration ranges from 0.1 mg/mL to 20.0 mg/mL, the most preferred being 5.0 mg/mL. Suitable buffering agents include histidine and sodium phosphate, ranging in concentration from 5 mM to 40 mM; with 10 mM preferred for sodium phosphate and 20 mM preferred for histidine, with sodium phosphate being the preferred buffering agent.
The compositions of the present invention may be either frozen, liquid, or lyophilized.
DETAILED DESCRIPTION OF THE INVENTION
In developing an appropriate drug dosage formulation, various factors are considered, including the solubility of a particular protein, its stability, and any particular handling requirements associated with the protein. While not all proteins are sensitive to handling, Applicants find that IL-11 is, in fact, sensitive to handling; both soluble aggregate and precipitate formation is observed after the protein has been “handled.” Such handling includes, for example, any of the usual normal shear forces associated with shipping to clinical sites and with packaging operations. Also, sometimes protein formulations are pumped through stainless steel or other tubing during manufacture or stressed by the delivery systems which can subject them to shear forces; sometimes it is necessary to subject the protein to a variety of freeze/thaw cycles, thereby also exposing the protein to potential denaturations.
While some efforts have been undertaken to overcome the stability problem by lyophilizing the protein of interest and shipping the protein in lyophilized form, once the protein has reached its destination it must be reconstituted by either the health care worker or the patient. Proper reconstitution requires that the procedure be done under sterile conditions, that it be done gently, and that an assessment be made regarding the integrity of the reconstituted solution. Because of potential inconveniences associated with the use of lyophilized dosage forms, when possible, liquid dosage forms are more desirable. Development of stable liquid dosage forms of protein pharmaceuticals is a challenge, because generally the liquid form is less stable than the lyophilized form. However, if the stability problems of a liquid dosage form are overcome, the liquid form can be utilized.
Applicants find that some of the chemical instability of IL-11 is a result of hydrolysis between Asp
133
and Pro
134
. Also, deamidation of Asn
49
to Asp
49
is detected. In addition, oxidation of Met
58
is observed. All of these chemical reactions are evidence of IL-11 protein chemical instability. IL-11 is also subject to certain physical instabilities including a dimerization process (which is actually a shift in equilibrium between the monomeric and dimeric forms of IL-11), as well as aggregate formation.
According to the present invention, the addition of glycine, at an appropriate pH, acts to prevent aggregation of IL-11 and protects IL-11 from the harmful effects of shearing. This in turn increases the ability to handle the protein and provides enhanced shelf-life for IL-11 products. The present invention also provides for IL-11 formulations, containing glycine, which are suitable for sub cutaneous injection. The IL-11 concentration ranges from 0.5 to 20.0 mg/mL. The IL-11 formulation can be in either a liquid or a lyophilized dosage form. Moreover, addition of an appropriate buffering agent slows the rate of hydrolysis, deamidation, and oxidation. Buffering is accomplished with a suitable buffer, or any buffering agent as is known to one skilled in the art, which will adequately buffer at neutral pH. Preferred is histidine; most preferred is sodium phosphate.


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Paul et

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