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Title:  Method of treating inflammatory bowel disease using a topical formulation of IL-11

United States Patent:  6,540,993

Issued:  April 1, 2003

Inventors:  Warne; Nicholas W. (Andover, MA); Bedrosian; Camille L. (Belmont Hills, MA); Keith, Jr.; James C. (Andover, MA); Schwerschlag; Ullrich S. (Beverly Farms, MA); Schendel; Paul F. (Wayland, MA)

Assignee:  Wyeth (Madison, NJ)

Appl. No.:  662994

Filed:  September 15, 2000

Abstract

Provided by the present invention are topical formulations of Interleukin-11 and methods for treating a variety of disorders, including inflammatory bowel diseases (e.g., Crohn's disease, ulcerative colitis, indeterminate colitis, and infectious colitis), mucositis (e.g., oral mucositis, gastrointestinal mucositis, nasal mucositis, and proctitis), necrotizing enterocolitis, inflammatory skin disorders (e.g., psoriasis, atopic dermatitis, and contact hypersensitivity), aphthous ulcers, pharyngitis, esophagitis, peptic ulcers, gingivitis, periodontitis, and ocular diseases (e.g., conjunctivitis, retinitis, and uveitis).

DETAILED DESCRIPTION OF THE INVENTION

Provided by the present invention are methods and compositions for topical administration of IL-11, for the treatment of various disorders, such as inflammatory bowel diseases (e.g., Crohn's disease, ulcerative colitis, indeterminate colitis, and infectious colitis), mucositis (e.g., oral mucositis, gastrointestinal mucositis, nasal mucositis, and proctitis), necrotizing enterocolitis, aphthous ulcers, psoriasis, pharyngitis, esophagitis, peptic ulcers, gingivitis, periodontitis, and ocular diseases (e.g., conjunctivitis, retinitis, and uveitis).

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods and materials are described. For purposes of the present invention, the following terms are defined below.

As used herein, the term "topical" refers to application at the site of inflammation (i.e., non-systemic administration), and includes, for example, oral, intranasal, intrabronchial, topical, and rectal routes of administration. The term "topical formulation" refers to a pharmaceutical formulation which is suitable for topical administration of the active ingredient.

As used herein, the term "pharmaceutically acceptable" refers to a carrier medium which does not interfere with the effectiveness of the biological activity of the active ingredient(s) and which is not toxic to the host to which it is administered.

As used herein, the terms "therapeutically effective amount" and "therapeutically effective dose" as applied to the active ingredient refers to the amount of the component in the composition or administered to the host that results in an increase in the therapeutic index of the host. The "therapeutic index" can be defined for purposes herein in terms of efficacy, i.e., extent of reduction or inhibition of inflammation. Suitable doses of the active ingredient can be determined using well-known methods, a variety of which are known and readily available in the pharmaceutical sciences, including, for example, measurement of markers associated with the disorder (e.g., TNF-.alpha., IL-1.beta., IL-6, and/or IL-12 p40 mRNA), the biological effects of TNF-.alpha., and decreased symptomatology.

Interleukin 11 (IL-11) is a pleiotropic cytokine that stimulates primitive lymphohematopoietic progenitor cells and synergizes with other hematopoietic growth factors to stimulate the proliferation and maturation of megakaryocytes. IL-11 is described in detail in International Application PCT/US90/06803, published May 30, 1991, as well as in U.S. Pat. No. 5,215,895; issued Jun. 1, 1993. A cloned human IL-11 was previously deposited with the ATCC, 12301 Parklawn Drive, Rockville, Md., on Mar. 30, 1990 under ATCC No. 68284. Moreover, as described in U.S. Pat. No. 5,270,181, issued Dec. 14, 1993, and U.S. Pat. No. 5,292,646, issued Mar. 8, 1994, IL-11 may also be produced recombinantly as a fusion protein with another protein. IL-11 can be produced in a variety of host cells by resort to now conventional genetic engineering techniques. In addition, IL-11 can be obtained from various cell lines, for example, the human lung fibroblast cell line, MRC-5 (ATCC Accession No. CCL 171), and Paul et al., the human trophoblastic cell line, TPA30-1 (ATCC Accession No. CRL 1583). A cDNA encoding human IL-11, as well as the deduced amino acid sequence (amino acids 1 to 199), is described in Proc. Natl. Acad. Sci. USA 87:7512 (1990). U.S. Pat. No. 5,292,646, supra, describes a des-Pro form of IL-11 in which the N-terminal proline of the mature form of IL-11 (amino acids 22-199) has been removed (amino acids 23-199). As is appreciated by one skilled in the art, any form of IL-11 which retains IL-11 activity, such as variants through the substitution or deletion of amino acids, analogs and derivatives of IL-11, is useful according to the present invention. The disclosure of each of the above publications is hereby incorporated by reference for the contents thereof.

In addition to recombinant techniques, IL-11 may also be produced by known conventional chemical synthesis. Methods for constructing the polypeptides useful in the present invention by synthetic means are known to those of skill in the art. The synthetically constructed cytokine polypeptide sequences, by virtue of sharing primary, secondary, or tertiary structural and conformational characteristics with the natural cytokine polypeptides are anticipated to possess biological activities in common therewith. Such synthetically constructed cytokine polypeptide sequences or fragments thereof, which duplicate or partially duplicate the functionality thereof may also be used in the method of this invention. Thus, they may be employed as biologically active or immunological substitutes for the natural, purified cytokines useful in the present invention.

Modifications in the protein, peptide or DNA sequences of these cytokines or active fragments thereof may also produce proteins which may be employed in the methods of this invention. Such modified cytokines can be made by one skilled in the art using known techniques. Modifications of interest in the cytokine sequences, e.g., the IL-11 sequence, may include the replacement, insertion or deletion of one or more selected amino acid residues in the coding sequences. Mutagenic techniques for such replacement, insertion or deletion are well known to one skilled in the art. (See, e.g., U.S. Pat. No. 4,518,584.)

Other specific mutations of the sequences of the cytokine polypeptides which may be useful therapeutically as described herein may involve, e.g., the insertion of one or more glycosylation sites. An asparagine-linked glycosylation recognition site can be inserted into the sequence by the deletion, substitution or addition of amino acids into the peptide sequence or nucleotides into the DNA sequence. Such changes may be made at any site of the molecule that is modified by addition of O-linked carbohydrate. Expression of such altered nucleotide or peptide sequences produces variants which may be glycosylated at those sites.

Additional analogs and derivatives of the sequence of the selected cytokine which would be expected to retain or prolong its activity in whole or in part, and which are expected to be useful in the present method, may also be easily made by one of skill in the art. One such modification may be the attachment of polyethylene glycol (PEG) onto existing lysine residues in the cytokine sequence or the insertion of one or more lysine residues or other amino acid residues that can react with PEG or PEG derivatives into the sequence by conventional techniques to enable the attachment of PEG moieties.

Additional analogs of these selected cytokines may also be characterized by allelic variations in the DNA sequences encoding them, or induced variations in the DNA sequences encoding them. It is anticipated that all analogs disclosed in the above-referenced publications, including those characterized by DNA sequences capable of hybridizing to the disclosed cytokine sequences under stringent hybridization conditions or non-stringent conditions (Sambrook et al., Molecular Cloning. A Laboratory Manual, 2d edit., Cold Spring Harbor Laboratory, New York (1989)) will be similarly useful in this invention.

Also considered as derivatives useful in these methods are fusion molecules, prepared by fusing the sequence or a biologically active fragment of the sequence of one cytokine to another cytokine or proteinaceous therapeutic agent, e.g., IL-11 fused to IL-6 (see, e.g., methods for fusion described in PCT/US91/06186 (WO92/04455), published Mar. 19, 1992). Alternatively, combinations of the cytokines may be administered together according to the method.

Thus, where in the description of the methods of this invention IL-11 is mentioned by name, it is understood by those of skill in the art that IL-11 encompasses the protein produced by the sequences presently disclosed in the art, as well as proteins characterized by the modifications described above yet which retain substantially similar activity. Standard laboratory tests are utilized to monitor progress of the treatment. Levels of TNF-.alpha. in serum or the biologic effects of TNF-.alpha. could be followed in a variety of these diseases. Decreased symptomatology could also be used to monitor the effectiveness of treatment as is well known to physicians skilled in the art of treating such disorders. Treatment is preferably prophylactic, but may also be at the onset of symptoms associated with the aforementioned disorders.

In one aspect of the invention, IL-11 is used in a therapeutic composition to alleviate or prevent the onset of symptoms associated with an inflammatory disorder. While it is possible to administer IL-11 alone, it is believed preferable to present it as part of a pharmaceutical formulation. In accordance with this aspect of the invention, the pharmaceutical compositions comprise IL-11 in a therapeutically effective dose together with one or more pharmaceutically acceptable carriers and optionally other therapeutic ingredients. A wide variety of pharmaceutically acceptable carriers are known to those of skill in the art. See, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 17th ed. (1985), which is incorporated by reference herein. Preferred carriers include inert, non-toxic solids (e.g., dextrose, dextrin, cellulose, pectin, starch, lactose, sucrose, and calcium phosphate,), semi-solids (e.g., glycerol stearate, polyethylene glycol, stearic acid, agar, gelatin, and propylene glycol) and liquids (e.g., buffered saline, water, an organic solvent, and pharmaceutically acceptable oils or fats).

As will be appreciated by those of skill in the art, the preferred form of the pharmaceutical composition of IL-11 will depend on the intended mode of administration, which in turn will depend on the location and nature of the inflammatory disorder to be treated. For example, delivery to the mouth (e.g., for treatment of oral mucositis, aphthous ulcers, gingivitis, periodontitis), head and/or neck (e.g., for treatment of pharyngitis, esophagitis) can be in the form of aqueous-based oral solutions, suspensions, emulsions, syrups, elixirs, gels, patches, lozenges, tablets, or capsules. Delivery to the gastrointestinal tract (e.g., for treatment of gastrointestinal mucositis, peptic ulcers and inflammatory bowel diseases, such as Crohn's disease, ulcerative colitis, indeterminate colitis, and infectious colitis) can be in the form of oral solutions, gels, suspensions, tablets, capsules, and the like. IL-11, either alone or in combination with other components in a pharmaceutical composition, can be added to infant formula for delivery to the gastrointestinal tract of an infant suffering from or susceptible to necrotizing enterocolitis. It is also possible to formulate the IL-11 preparation for rectal administration (e.g., for treatment of proctitis), e.g., in the form of enema, suppositories, rectal-foam, and the like. Delivery to the eye (e.g., for treatment of ocular diseases such as conjunctivitis, retinitis, and uveitis) can be in the form of solutions, gels, or suspensions. Delivery to the nose (e.g., for treatment of nasal mucositis) can be in the form of solutions, gels, or suspensions. The intranasal formulations may be formulated, for example, into an aqueous or partially aqueous solution, which can then be utilized in the form of a nasal drop or an aerosol. Delivery to the skin (e.g., for treatment of psoriasis) can be in the form of aqueous-based solutions, gels, suspensions, lotions, creams, ointments, patches, and the like.

Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators. Suitable examples of liquid carriers for oral administration include water (partially containing additives as above), alcohols (including monohydric alcohols and polyhydric alcohols, e.g., glycols) and their derivatives, oils (e.g., peanut oil, sesame oil, olive oil, and coconut oil), and combinations of the above. Compositions comprising such carriers and adjuvants may be formulated using well known conventional materials and methods. Such materials and methods are described, for example, in Remington's Pharmaceutical Sciences, supra. Infant formula is also a suitable liquid carrier, particularly when the pharmaceutical composition is used to treat or prevent necrotizing enterocolitis.

A solid carrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablet preferably contain up to 99% of the active ingredient, and may be formulated for immediate and/or sustained release of the active ingredient. Suitable solid carriers include, for example, calcium or sodium phosphate, magnesium stearate, talc, sugars, glycine, lactose, dextrin, starch, gelatin, cellulose, cellulose derivatives (e.g., methyl cellulose, hydroxypropylmethyl cellulose, and sodium carboxymethyl cellulose), polyvinylpyrrolidone, low melting point waxes, and combinations of the above.

Oral tablets may be prepared using a variety of well known methods and in a variety of conventional forms. Exemplary forms include dry powder compaction tablets, micro-particulate systems (e.g., wherein the active ingredient is spray-dried onto a scaffold particle), and hard or soft-gel capsules. The tablets may be optionally covered with an enteric coating, which remains intact in the stomach, but will dissolve and release the contents of the tablet once it reaches the small intestine. Most currently used enteric coatings are those which remain undissociated in the low pH environment of the stomach, but readily solubilize when the pH rises to about 4 or 5. A number of commercially available enteric coatings may be used depending on the target part of the intestinal tract, i.e., the site of the inflammatory bowel disorder in the patient. Such coatings include, for example, methacrylic acid-methacrylic acid ester-based copolymer, which is sold under the trade name "Eudragit"; anionic water-soluble, polymer cellulose ether, which is sold under the trade name "Aqualon"; cellulose acetate phthalate; polyvinyl acetate phthalate; hydroxypropyl methylcellulose phthalate; and the like. Compositions comprising such carriers and adjuvants may be formulated, and tablets prepared from such compositions, using well known conventional materials and methods. Such materials and methods are described, for example, in Remington's Pharmaceutical Sciences, supra.

In one embodiment of the invention, the pharmaceutical composition comprises one or more sustained or controlled release excipients such that a slow or sustained, preferably constant, release of the active ingredient is achieved. A wide variety of suitable excipients are known in the art and are not a part of this invention. Such sustained/controlled release excipients and systems are described, for example, in U.S. Pat. No. 5,612,053 (Baichwal et al.), U.S. Pat. No. 5,554,387 (Baichwal), U.S. Pat. No. 5,512,297 (Baichwal), U.S. Pat. No. 5,478,574 (Baichwal et al.), and U.S. Pat. No. 5,472,711 (Baichwal et al.), each of which is incorporated by reference herein. If desired, the pharmaceutical composition can be formulated to provide a pulse dose of the active ingredient. A variety of pulse-dose systems, which provide low or high-pulsed doses, are known in the art and are not a part of this invention. Such pulse-dose formulations are described, for example, in Y. H. Bae, "Stimuli-Sensitive Drug Delivery," Controlled Drug Delivery (ed. K. Park) ACS Press, Washington, D.C. (1997); Drugs (1982) 23:207-226; Fertil.Steril. (1983) 39:695-699, 1983; Brit. J. Cancer (1982) 45:86-94; Int. J. Radiat. Oncol. Biol. Phys. (1982) 8:915-919; J. Clin. Endocrinol. Metab. (1981) 53:184-91; and Diabetes (1977) 26:571-581.

In another embodiment of the invention, the pharmaceutical composition is formulated to provide direct and/or targeted delivery of the active ingredient to a specific anatomic site or sites within the gastrointestinal tract; e.g., the duodenum, jejunum, ileum, cecum and/or colon, each of which is believed to contain receptors for IL-11. Methods for providing targeted delivery of macromolecules, including proteins and polypeptides, to specific tissues or organs within a mammalian host are well known in the art and not a part of this invention. Such targeted delivery systems are described, for example, in R. J. Mrsny, "Site-Specific Drug Delivery in the Gastrointestinal Tract," Controlled Drug Delivery, supra; M. Mezei and D. Meisner, "Liposomes and Nanoparticles as Ocular Drug Delivery Systems," Biopharmaceutics of Ocular Drug Delivery, Ch. 6 (ed. P. Edman) CRC Press, Boca Raton, Fla. (1992); T. L. Bowersock and H. Hogenesch, "Oral Immunization Using Microparticles," Controlled Drug Delivery, supra; M. Mezei, "Liposomes in Topical Application of Drugs," Liposomes as Drug Carriers: Trends and Progress (ed. G. Gregoriadis), J. Wiley & Sons Publ. USA (1988), pp. 663-677; and J. J. Berti and J. J. Lipsky, "Transcutaneous Drug Delivery: A Practical Review," Mayo CLW Proc. (1995) 70:581-586.

IL-11, either alone or in combination with other therapeutic agents, may also be administered topically in the form of a dermal patch or transdermal delivery system. Such patches and systems are especially suited for treatment of inflammatory skin disorders such as psoriasis, and for inflammatory disorders involving the mucosa of the head, neck and/or mouth, e.g., oral mucositis, pharyngitis, esophagitis, gingivitis, perodontitis, and aphthous ulcers. In this embodiment of the invention, the pharmaceutical composition may be administered through the use of a dermal patch containing the active ingredient(s) and a carrier that is inert to the active ingredient(s), non-toxic to the skin or mucosal epithelium, and allows delivery of the agent to the dermis and/or epithelium. Dermal patches and delivery systems, utilizing active or passive transdermal delivery carriers, comprising IL-11 may be prepared suing well known methods and materials, including, for example, microporous membranes, silicon polymers and diffusion matrixes. Such materials and methods are described, for example, in Remington's Pharmaceutical Sciences, supra.

In yet another embodiment of the invention, at least one proteinase inhibitor is combined with IL-11 in a therapeutic composition to treat inflammatory diseases. In accordance with this aspect of the invention, the pharmaceutical composition comprises IL-11 and one or more proteinase inhibitors, which stabilize the IL-11 molecule against proteolytic degradation. It may be preferable to include a proteinase inhibitor in the pharmaceutical composition to protect IL-11 against proteolytic enzymes in the gastrointestinal tract, particularly in rectal formulations. A wide variety of suitable proteinase inhibitors are known in the art and are not a part of this invention. Suitable proteinase inhibitors include, for example, aprotinin, a kallikrein-trypsin inhibitor, and protein serine proteinase inhibitors such as .alpha.-macroglobulin, soybean trypsin inhibitor, and ovomucoid.

A suitable treatment regimen for patients undergoing treatment, including for example prophylactic treatment, may be determined by the attending physician based upon such factors as the patient's age, sex, weight, and general health. Generally, a suitable dose of IL-11 ranges broadly, preferably between about 1 and about 250 microgram (.mu.g) per kilogram (kg) of body weight of recipient per treatment. Another suitable dose may be in the range of about 1 to about 100 gg per kg of body weight, and more preferably in the range of about 10 to about 50 .mu.g per kg of body weight. If desired, these doses can be adjusted to units. A unit is conventionally described as the concentration of polypeptide which leads to half-maximal stimulation in a suitable assay, e.g., for IL-11, the T1165 assay described in PCT/US90/06803. Doses may be administered daily for between one day and six months, or for as long as is deemed necessary and safe, as is readily ascertained by standard tests by the attending physician, depending upon the nature of the disorder being treated. Where appropriate, the dosages may be adjusted upward or downward, for example, a dosing regimen requiring administration of IL-11 at a dose of 25 .mu.g/kg, daily for one week, or fewer days, or multiple weeks if indicated. The progress of treatment is appropriately monitored by measurement of markers associated with the disorder being treated to determine if such a dose results in a decrease of for example, TNF-.alpha. levels (or corresponding marker) and if not, increasing the dose two-fold for an additional time period of treatment and measurement of marker levels until an effective dosing regimen is reached.

Claim 1 of 12 Claims

We claim:

1. A method of treating Inflammatory Bowel Disease comprising administering to a patient a topical formulation of a pharmaceutical composition composed of a pharmaceutically effective amount of IL-11 and a carrier for delivery of IL-11 to the gastrointestinal tract.
 


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