U.S. patent application number 11/050246 was filed with the patent office on 2005-06-16 for topical formulation for delivery of interleukin-11.
Invention is credited to Bedrosian, Camille L., Keith, James C. JR., Schendel, Paul F., Schwertschlag, Ullrich S., Warne, Nick W..
Application Number | 20050129658 11/050246 |
Document ID | / |
Family ID | 34519601 |
Filed Date | 2005-06-16 |
United States Patent
Application |
20050129658 |
Kind Code |
A1 |
Warne, Nick W. ; et
al. |
June 16, 2005 |
Topical formulation for delivery of interleukin-11
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).
Inventors: |
Warne, Nick W.; (Andover,
MA) ; Bedrosian, Camille L.; (Belmont Hills, MA)
; Keith, James C. JR.; (Andover, MA) ;
Schwertschlag, Ullrich S.; (Beverly Farms, MA) ;
Schendel, Paul F.; (Wayland, MA) |
Correspondence
Address: |
MINTZ, LEVIN, COHN, FERRIS, GLOVSKY
AND POPEO, P.C.
ONE FINANCIAL CENTER
BOSTON
MA
02111
US
|
Family ID: |
34519601 |
Appl. No.: |
11/050246 |
Filed: |
February 3, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11050246 |
Feb 3, 2005 |
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09604378 |
Jun 27, 2000 |
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6887461 |
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09604378 |
Jun 27, 2000 |
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09179026 |
Oct 26, 1998 |
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6126933 |
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09179026 |
Oct 26, 1998 |
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08892407 |
Jul 15, 1997 |
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5948402 |
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08892407 |
Jul 15, 1997 |
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08495724 |
Jun 27, 1995 |
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5679339 |
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Current U.S.
Class: |
424/85.2 |
Current CPC
Class: |
A61K 38/2073 20130101;
A61K 9/0031 20130101; A61K 9/006 20130101; A61K 9/0056
20130101 |
Class at
Publication: |
424/085.2 |
International
Class: |
A61K 038/20 |
Foreign Application Data
Date |
Code |
Application Number |
May 30, 1996 |
WO |
PCT/US96/08059 |
Claims
What is claimed is:
1. A method of treating an inflammatory disorder, comprising
administering a topical formulation of a pharmaceutical composition
comprising a pharmaceutically effective amount of IL-11.
2. The method of claim 1 wherein the disorder is mucositis.
3. The method of claim 2 wherein the mucositis is oral
mucositis.
4. The method of claim 2 wherein the mucositis is gastrointestinal
mucositis.
5. The method of claim 1 wherein the disorder is an inflammatory
skin disorder.
6. The method of claim 5 wherein the inflammatory skin disorder is
psoriasis.
7. The method of claim 1 wherein the disorder is necrotizing
enterocolitis.
8. The method of claim 1 wherein the disorder is an ocular
disease.
9. The method of claim 8 wherein the ocular disease is selected
from the group consisting of conjunctivitis, retinitis and
uveitis.
10. The method of claim 1 wherein the disorder is selected from the
group consisting of aphthous ulcers, pharyngitis, esophagitis,
peptic ulcers, gingivitis, and periodontitis.
11. The method of claim 1 wherein the pharmaceutically effect
amount of IL-11 is between about 1 and about 250 .mu.g/kg body
weight.
12. The method of claim 1, wherein the pharmaceutical composition
comprises a solution containing IL-11 and a suitable liquid
carrier.
13. The method of claim 12, wherein the suitable liquid carrier is
selected from the group consisting of water, organic solvents, oils
and fats.
14. The method of claim 12, wherein the suitable liquid carrier is
sodium bicarbonate.
15. The method of claim 12, wherein the suitable liquid carrier is
an infant formula.
16. The method of claim 1, wherein the pharmaceutical composition
comprises an immediate release carrier for immediate release of
IL-11 into the oral cavity.
17. The method of claim 16, wherein the immediate release carrier
is selected from the group consisting of sugars, glycine, lactose,
dextrin, starch, gelatin, cellulose, methyl cellulose,
hydroxypropylmethyl cellulose, and sodium carboxymethyl
cellulose.
18. The method of claim 16, wherein the topical formulation is
selected from the group consisting of an oral gel, tablet or
suspension.
19. The method of claim 1, wherein the pharmaceutical composition
comprises an immediate release carrier for delivery of IL-11 to the
gastrointestinal tract.
20. The method of claim 19, wherein the immediate release carrier
is selected from the group consisting of sugars, glycine, lactose,
dextrin, starch, gelatin, cellulose, methyl cellulose,
hydroxypropylmethyl cellulose, and sodium carboxymethyl
cellulose.
21. The method of claim 19, wherein the topical formulation is a
pill, tablet or capsule.
22. The method of claim 1, wherein the pharmaceutical composition
comprises a sustained-release carrier for delivery of IL-11 to the
oral cavity.
23. The method of claim 21, wherein the topical formulation is a
patch, lozenge or an uncoated tablet.
24. The method of claim 1, wherein the pharmaceutical composition
comprises a sustained-release carrier for delivery of IL-11 to the
gastrointestinal tract.
25. The method of claim 24, wherein the topical formulation is a
pill, tablet or capsule.
26. The method of claim 1, wherein the pharmaceutical composition
comprises an immediate release carrier for delivery of IL-11 for
cervical administration.
27. The method of claim 25, wherein the topical formulation is
selected from the group consisting of a topical cream, solution,
ointment, and gel.
28. The method of claim 1, wherein the pharmaceutical composition
comprises a sustained-release carrier for delivery of IL-11 for
cervical administration.
29. The method of claim 26, wherein the topical formulation is a
topical cream, solution, ointment or gel.
30. The method of claim 1, wherein the pharmaceutical composition
comprises an enema preparation of IL-11 and a suitable liquid
carrier for delivery to the colon.
31. The method of claim 1, wherein the pharmaceutical composition
comprises a proteinase inhibitor.
32. The method of claim 31, wherein the proteinase inhibitor is
selected from the group consisting of aprotinin,
.alpha.-macroglobulin, soybean trypsin inhibitor, and
ovomucoid.
33. The method of claim 31, wherein the proteinase inhibitor is
aprotinin.
34. The method of claim 1, wherein the topical formulation
comprises an enteric coating.
35. The method of claim 34, wherein the enteric coating is selected
from the group consisting of a methacrylic acid-methacrylic acid
ester-based copolymer, an anionic water-soluble polymer cellulose
ether, cellulose acetate phthalate, polyvinyl acetate phthalate,
and hydroxypropyl methylcellulose phthalate.
36. A composition for treating an inflammatory disorder, wherein
said composition is a topical formulation comprising a
pharmaceutically effective amount of IL-11 and a suitable liquid
carrier.
37. The composition of claim 36, wherein said suitable liquid
carrier is sodium bicarbonate.
38. The composition of claim 36, wherein said suitable liquid
carrier is an infant formula.
39. A composition for treating an inflammatory disorder, wherein
said composition is a topical formulation comprising a
pharmaceutically effective amount of IL-11 and a suitable solid
carrier.
40. The composition of claim 39, wherein said suitable solid
carrier is an immediate release carrier.
41. The composition of claim 40, wherein said immediate release
carrier is selected from the group consisting of sugars, glycine,
lactose, dextrin, starch, gelatin, cellulose, methyl cellulose,
hydroxypropylmethyl cellulose, and sodium carboxymethyl
cellulose.
42. The composition of claim 39, wherein said suitable solid
carrier is a sustained-release carrier.
43. The composition of claim 39, wherein said suitable solid
carrier comprises a proteinase inhibitor.
44. The composition of claim 39, wherein the proteinase inhibitor
is selected from the group consisting of aprotinin,
a-macroglobulin, soybean trypsin inhibitor, and ovomucoid.
45. The composition of claim 44, wherein the topical formulation
comprises an enteric coating.
46. The composition of claim 45, wherein the enteric coating is
selected from the group consisting of a methacrylic
acid-methacrylic acid ester-based copolymer, an anionic
water-soluble polymer cellulose ether, cellulose acetate phthalate,
polyvinyl acetate phthalate, and hydroxypropyl methylcellulose
phthalate.
Description
RELATED APPLICATION(S)
[0001] This is a Continuation of U.S. application Ser. No.
09/179,026, filed Oct. 26, 1998, which is a continuation-in-part of
U.S. application Ser. No. 08/892,407, filed Jul. 15, 1997, which is
a divisional of U.S. application Ser. No. 08/495,724, filed Jun.
27, 1995, now Pat. No. 5,679,339, issued Oct. 21, 1997, the entire
teachings of which are incorporated herein by reference.
FIELD OF INVENTION
[0002] The present invention relates generally to novel
compositions and methods for topical delivery of interleukin-11
(IL-11). In preferred embodiments, patients are treated employing
topical delivery of recombinant human IL-11 for 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).
BACKGROUND OF THE INVENTION
[0003] Inflammatory responses include a broad range of host
reaction to a variety of insults, such as injury, infection, or
rejection. It is the overproduction of mediators that is believed
to be associated with a broad range of disorders, including AIDS,
arthritis (rheumatoid arthritis, osteoarthritis,
spondyloarthropathies), antibiotic-induced diarrheal diseases,
multiple sclerosis, osteoporosis, gingivitis, peptic ulcer disease,
esophagitis, diabetes, retinitis, uveitis, reperfusion injury after
myocardial infarction, cerebral vascular accident, aphthous ulcers
(oral), atherosclerosis, tumor metastases, asthma, preeclampsia,
pancreatitis, psoriasis, infertility and allergic disorders such as
rhinitis, conjunctivitis, and urticaria.
[0004] Some of these disorders and their symptoms are briefly
summarized below. According to the methods of the present
invention, IL-11 is administered topically to modulate the host's
over reaction at the site of insult, thereby treating the following
disorders.
[0005] Inflammatory Bowel Disease: Inflammatory bowel disease is
the term generally applied to four diseases of the bowel, namely
Crohn's disease, ulcerative colitis, indeterminate colitis, and
infectious colitis.
[0006] Ulcerative Colitis: Ulcerative colitis is a chronic
inflammatory disease of unknown etiology afflicting the large
intestine. The course of the disease may be continuous or
relapsing, mild or severe. The earliest lesion is an inflammatory
infiltration with abscess formation at the base of the crypts of
Lieberkuhn. Coalescence of these distended and ruptured crypts
tends to separate the overlying mucosa from its blood supply,
leading to ulceration. Signs and symptoms of the disease include
cramping, lower abdominal pain, rectal bleeding, and frequent,
loose discharges consisting mainly of blood, pus, and mucus with
scanty fecal particles. A total colectomy may be required for acute
severe or chronic, unremitting ulcerative colitis.
[0007] Crohn's Disease: Crohn's disease (also known as regional
enteritis or ulcerative ileitis) is also a chronic inflammatory
disease of unknown etiology but, unlike ulcerative colitis, it can
affect any part of the bowel. The most prominent feature of the
disease is the granular, reddish-purple edematous thickening of the
bowel wall. With the development of inflammation, these granulomas
often lose their circumscribed borders and integrate with the
surrounding tissue. Diarrhea and obstruction of the bowel are the
predominant clinical features. As with ulcerative colitis, the
course of the disease may be continuous or relapsing, mild or
severe but, unlike ulcerative colitis, it is not curable by
resection of the involved segment of bowel. Most patients with
Crohn's disease require surgery at some point, but subsequent
relapse is common and continuous medical treatment is usual.
[0008] Mucositis: Mucositis involves ulcerative breakdown of
mucosal epithelial tissue, and is literally defined as inflammation
of the mucous membrane. The pathophysiology of mucositis in
response to toxic insults to the mucosa by chemotherapy or by
ionizing radiation is complex and involves a cascade of
interactions among cells, cytokines and the oral microflora. The
underlying premise for susceptibility of the mucosa of the
oropharynx and gastrointestinal tract to chemotherapy or radiation
damage is related to rapid epithelial stem cell turnover. Mucositis
may be characterized by the following phases:
[0009] 1. Early inflammatory phase characterized by release of
inflammatory cytokines in response to local tissue damage caused by
cytotoxic agent(s);
[0010] 2. Epithelial phase characterized by death of basal cells,
which hinders re-population of the epithelium. This inability to
regenerate leads to atrophy followed by ulceration. The ulceration
represents loss of an important anatomic barrier at a site of local
microflora;
[0011] 3. Infection phase characterized by local invasion of
microflora that results in an inflammatory response to the local
infection. The inflammation results in additional local tissue
damage and possibly erosive ulceration; and
[0012] 4. Healing phase characterized by resolution of the
infection and regeneration of epithelium.
[0013] Oral mucositis produces the following clinical symptoms and
signs resulting from cellular damage: 1) sensation of dryness; 2)
asymptomatic redness and erythema; 3) solitary white elevated
desquamative patches which are painful upon pressure contact; and
4) large, painful, contiguous pseudomembranous lesions associated
with dysphagia and decreased oral intake. These spontaneously
painful lesions histopathologically show loss of epithelial cells
to the basement membrane, which exposes the connective tissue
stroma with its associated innervation.
[0014] As with oral mucosa, gastrointestinal mucosal damage results
from disturbance of cellular mitosis that leads to reduction in the
turnover rate of the basal cells of the intestinal crypts. The
symptoms and signs of gastrointestinal mucositis include tenesmus
(painful ineffectual straining at stool), pain, bleeding, diarrhea,
telangectasia (neovascularization), and progression to ulceration.
Early signs of diarrhea include increased stool frequency, loose or
watery stool, food aversion, increased bowel sounds, abdominal
pain, and some loss of skin turgor indicative of dehydration. When
the diarrhea is severe it may be associated with mucosal
ulceration, bleeding, intestinal perforation and proctitis. Stool
exam may reveal occult blood and fecal leukocytes.
[0015] Necrotizing Enterocolitis: Necrotizing enterocolits is an
inflammatory disease of unknown etiology that afflicts between 1-5%
of all infants admitted to neonatal intensive care units, most of
whom are premature infants. Signs and symptoms include abdominal
distention, gastrointestinal hemorrhage, and feeding intolerance.
The disease most often involves the ileum and colon, and is
characterized by loss of epithelium and submucosal edema,
ulcerations, and, in severe cases, transmural necrosis.
[0016] Psoriasis: Psoriasis is a chronic inflammatory skin disorder
involving hyperproliferation of the epidermis and inflammation of
both the epidermis and the dermis. Macrophage, T-cells and
neutrophil infiltration of the dermis and epidermis is seen, and
proinflammatory mediators are released from these activated
cells.
[0017] Aphthous Ulcers (oral): Although the cause of aphthous
ulcers remain unknown, many physicians believe they are caused by
autoimmune phenomena, which cause the destruction of discrete areas
of the oral mucosa which leads to oral ulceration. Among the
cytokines present in these active areas of ulceration, TNF-.alpha.
appears to play a predominant role.
[0018] Gingivitis: Adult periodontitis is strongly associated with
infection by Porphyromonas gingivalis. Proteolytic enzymes, which
are produced in large quantity by this bacteria, are considered as
important pathogenic agents. The increased production and flow of
gingival crevicular fluid (GCF) is an important change in gingival
tissues during periodontal infection, correlating with clinical
indices of gingival inflammation. Indeed, salivary protein and
albumin concentrations of individuals with periodontitis, which are
an indication of plasma leakage due to vascular permeability
enhancement (VPE), are significantly increased compared to healthy
subjects. The production of GCF appears dependent on VPE induced at
periodontitis sites, presumably involving proteinase(s) of P.
gingivalis in their generation.
[0019] Esophagitis: The most common cause of esophagitis is the
chronic reflux of hydrochloric acid from the stomach due to
inefficiency of the cardiac sphincter of the stomach. The chronic
presence of acid in the lower esophagus leads to damage of the
esophageal mucosa. In the most severe form, a syndrome called
Barrett's esophagus can develop which often leads to esophageal
cancer. Other causes of esophagitis include parenteral chemotherapy
and ionizing radiation, associated with radiation therapy for
cancer in the thoracic cavity.
[0020] Retinitis: Inflammation of the light sensitive retina,
retinitis, can occur due to a variety of viral, bacterial or
autoimmune etiologies. The end result is destruction of the retina
and loss of sight.
[0021] Uveitis: Inflammation of the anterior portion of the eye its
associated structures, the iris and cornea occurs with a relatively
high frequency in patients with autoimmune disorders.
[0022] Peptic Ulcer Disease: Inhibition of gastric acid secretion
with H.sub.2-receptor antagonists and, more recently, blockers of
H.sup.-,K.sup.--ATPase (also known as the proton pump) has been the
mainstay of therapy for peptic ulcer disease. The pathophysiology
of peptic ulcers remains obscure. An appreciation of the complexity
of the physiology of the gastric mucosa has led to a hypothesis
that peptic ulcers are the result of an imbalance in the relative
importance of aggressive (acid, pepsin) and protective (mucus,
bicarbonate, blood flow, prostaglandins, etc.) factors. Infection
of the mucosa of the human gastric antrum with the bacterium
Helicobacter pylori has been widely accepted as the cause of
chronic, active, type B gastritis. Further, this form of gastritis
has been linked directly to peptic ulcer disease by studies showing
that eradication of H. pylori reverses this gastritis and prevents
duodenal ulcer relapse. Because cytokines are the principal
mediators by which immune/inflammatory cells communicate with each
other and with other cells, it is likely that these small peptides
are involved in the pathogenesis of chronic active type B gastritis
and the resulting peptic ulcer disease.
[0023] Some cytokines (IL-1, epidermal growth factor, transforming
growth factor-.alpha., acidic and basic fibroblast growth factors)
tip the balance towards peptic ulcer healing; others (tumor
necrosis factor-.alpha.) appear to have no effect; still others
(IL-4) may even cause gastrointestinal damage.
[0024] The infiltration and activation of multiple types of
inflammatory cells result in a series of degenerative changes in
the vasculature of the affected area, as well as inciting damage of
the surrounding parenchymal tissue.
SUMMARY OF THE INVENTION
[0025] Provided by the present invention are novel compositions for
the topical administration of IL-11 in order to treat a number of
disorders where such administration is preferable to systemic
administration. Among the reasons that local administration may be
preferred are the ease of administering a topical formulation
compared to administration of subcutaneous injectable formulations.
In certain classes of patients, the toxicity profile of
chemotherapeutic agents may be such that concurrent parenteral
administration of IL-11 is relatively unsuited. Other patients may
have medical conditions for which the adverse event profile of
parenteral IL-11 is relatively unsuited.
[0026] According to the present invention, IL-11, analogs, and
derivatives thereof, are administered to patients, either
prophylactically or at the onset of symptoms associated with the
aforementioned disorders, through one of several forms of local
administration described below. IL-11 can be administered in
suitable pharmaceutically acceptable carriers either alone or in
combination with other conventional agents useful in alleviating
the symptoms associated with the aforementioned disorders.
[0027] In one embodiment, the present invention comprises
preparations of IL-11 which are suitable for oral delivery to the
mouth. Suitable oral preparations may be prepared with
aqueous-based solutions such as sodium bicarbonate (e.g.,
Brioschi.RTM.), or in gels and suspensions for topical
administration in the mouth. Oral preparations may also take the
form of patches for delivery of IL-11 to the mouth via sustained
release. Additional oral preparations may comprise IL-11 in the
form of a lozenge or an uncoated tablet which is retained in the
mouth. The oral preparations are particularly well-suited for
disorders and inflammatory responses involving the mucosa of the
head, neck and/or mouth. Such conditions include oral mucositis,
pharyngitis, esophagitis, gingivitis, periodontitis, and aphthous
ulcers (oral). Such conditions may result, for example, from
chemotherapy or radiotherapy for head and neck cancer, cervical
esophageal cancer or lung cancer.
[0028] In other embodiments, the present invention comprises
preparations of IL-11 which are suitable for topical delivery for
mucosa and/or dermis. Such topical preparations may be prepared in
the form of aqueous-based solutions, gels, ointments or creams for
topical administration, as gels and suspensions for cervical
administration, as pills, tablets, capsules or suppositories for
immediate or sustained release to the gastrointestinal tract, or in
the form of solution for enema. Such topical preparations are
especially suited for treatment of disorders relating to local
regions, such as psoriasis, as well as inflammatory bowel disease,
esophagitis, and gastrointestinal mucositis. Some of these
conditions may result, for example, from chemotherapy and/or
radiotherapy for colorectal cancer, prostate cancer, cervical
esophageal cancer or lung cancer.
[0029] Suitable doses of IL-11 are generally in the range of
between about 1 and about 250 .mu./kg body weight, and may be
administered from once a week up to about six times daily.
Treatment may continue for a period of between one day and six
months, or for as long as is deemed necessary and safe in the
treatment of the aforementioned disorders, as is readily
ascertained by standard tests by the attending physician, depending
upon the nature of the disorder being treated.
DETAILED DESCRIPTION OF THE INVENTION
[0030] 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).
[0031] 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.
[0032] 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.
[0033] 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.
[0034] 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.
[0035] 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.
[0036] 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.
[0037] 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.)
[0038] 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.
[0039] 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.
[0040] 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.
[0041] 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/U.S.91/06186
(WO92/04455), published Mar. 19, 1992). Alternatively, combinations
of the cytokines may be administered together according to the
method.
[0042] 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.
[0043] 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).
[0044] 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.
[0045] 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.
[0046] 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.
[0047] 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.
[0048] 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.
[0049] 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.
[0050] 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.
[0051] 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.
[0052] 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 .mu.g 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.
[0053] The following examples illustrate the methods of the present
invention and in particular the use of topical IL-11 formulations
in treating Crohn's disease, chemotherapy-induced ulcerative
mucositis, colitis (colonic ulcers), psoriasis, and inflammatory
bowel disease. However, as the skilled artisan will recognize from
the present disclosure, numerous modifications and variations are
possible and comprised within the present invention. The examples
are not exhaustive, and do not limit the scope of the invention in
any way.
EXAMPLE 1
Treatment of Inflammatory Bowel Disease
[0054] In two well-established models it has been shown that
subcutaneous rhIL-11 is able to significantly reduce the clinical
signs and histologic lesions of gastrointestinal inflammation:
acetic acid-induced colitis, a model of acute injury; and the
HLA-B27 transgenic rat, a model of chronic inflammation. While
conducting further studies of the mechanism of rhIL-11 action, oral
topical activity was seen in hamsters with 5-FU induced oral
mucositis and mucosal activity was documented in in vitro/ex vivo
studies of intestinal transport. Subsequently, these models have
been used to confirm the effectiveness of topically and orally
administered rhIL-11.
[0055] Applicants first examined local topical delivery of rhIL-11
to the colon by enema. To determine the stability of rhIL-11 in the
lumen of the gut, rhIL-11 and intestinal chyme from HLA-B27 rats
were combined and incubated at 37.degree. C. for up to several
hours in the presence or absence of a proteinase inhibitors. Fluid
samples were analyzed periodically by SDS-PAGE. The addition of a
proteinase inhibitor, avian ovoinhibitor, stabilized rhIL-11
against the proteolytic degradation observed in the rat chyme.
Next, twenty-three HLA-B27 rats (22-23 weeks old) were randomized
to one of six treatment groups: rhIL-11 Vehicle; 50 .mu.g/ml
ovoinhibitor; rhIL-11 200 .mu.g/ml; or rhIL-11 at 50, 200 or 800
.mu./ml combined with 50 .mu.g/ml of ovoinhibitor. Animals received
a colonic irrigation (1 ml) of the respective formulations, Monday,
Wednesday and Friday, for three weeks. Stool character was assessed
daily. At the end of the experiment, terminal blood samples were
obtained, and then the animals were euthanized by an overdose of
anesthetic. Platelet counts and colonic mucosal lesions were
quantitated and analyzed as described previously. Platelet counts
were not different in any of the groups. In the vehicle treated
animals, normal, soft or diarrhea stools were present at 16.5, 33,
or 50% of the total animal observation days, respectively. In the
50 .mu.g/ml+50 .mu.g/ml ovoinhibitor animals, normal, soft, or
diarrhea stools were present at 65.9, 25, or 9.1% of the total
animal observation days, respectively. Histologic lesion scores
were 7.25.+-.1.4 and 7.8.+-.1.4 in the vehicle and ovoinhibitor
groups, respectively. ANOVA testing showed that all of the rhIL-11
lesion scores were lower than either vehicle or ovoinhibitor
(P<0.05), and the lowest lesion score was in the 50 .mu.g
rhIL-11+50 .mu.g ovoinhibitor group, 2.1.+-.0.4. Topically
administered rhIL-11 was beneficial in the model of chronic
inflammatory bowel disease when combined with a proteinase
inhibitor. Since this study was based around a dose of 200
.mu.g/ml, no dosing group of 50 .mu.g/ml without the proteinase
inhibitor was included.
[0056] Next, the effects of lower doses of recombinant human
interleukin eleven (rhIL-11) with or without Aprotinin (an FDA
approved and marketed Proteinase inhibitor) delivered by enema were
assessed in the acetic acid model of acute colitis. Colitis was
induced in the descending colons of 80 anesthetized Sprague-Dawley
rats by intraluminal instillation of 10% acetic acid for 10 seconds
on Day 0 of the study. The rats were randomly assigned to the 8
groups according to their different treatments as described
below:
1TABLE 1 Group No. Animals Dosage Schedule 1 Vehicle 10 1 ml.
Buffer 3 days/wk/2 wks. 2 Aprotinin 10 50 .mu.g/ml 3 days/wk/2 wks.
3 & 4 rhIL-11 .+-. 20 10 .mu.g/.mu.g/ml 3 days/wk/2 wks
Aprotinin 5 & 6 rhIL-11 .+-. 20 25 .mu.g/g/ml 3 days/wk/2 wks
Aprotinin 7 & 8 rhIL-11 .+-. 20 50 .mu.g/ml 3 days/wk/2 wks
Aprotinin
[0057] Peripheral hematologic parameters were assessed. On day 15,
the animals were euthanized, and colons were removed for
determination of lesion scores by gross examination and histologic
evaluation. Platelet counts in all groups were in normal range. All
animals treated with acetic acid developed diffuse colonic lesions,
characterized by ulceration, hemorrhage, edema, depletion of goblet
cells, and infiltration of leukocytes. The gross and histologic
lesions of all rhIL-11 treated animals were significantly reduced
compared with vehicle controls on day 15. Administration of rhIL-11
10 .mu.g with 50 .mu.g Aprotinin 3 days/week/for 2 weeks or 50
.mu.g/ml rhIL-11 3 days/wk/2 wks appeared to be most effective in
this model of colonic mucosal injury.
EXAMPLE 2
Treatment of Oral Mucositis
[0058] 95 male Golden Syrian hamsters were equally divided into
five groups. Group 1 [control] received phosphate buffered saline
and 0.5% hamster serum (vehicle) topically on days 3-14. Groups 2
and 4 received 50 .mu.g IL-11 subcutaneously, twice daily, on days
0-14 and 3-14, respectively. Groups 3 and 5 received 100 .mu.g
IL-11 topically, four times daily, on days 3-14 and 0-14,
respectively. Ulcerative mucositis was induced by administration of
60 mg/kg body weight of 5-fluorouracil on days 0 and 2. The left
buccal pouch was superficially irritated on day 4. Ulcerative
mucositis was evaluated blindly starting on day 6 by scoring
standardized photographs. The scoring was done on a scale of 0-10,
with 10 being the most severe. Animals were weighed daily. Blood
was taken from 3 animals per group on days 6, 10 and 14.
[0059] For the entire experiment, average mean ulcerative mucositis
scores for all groups were significantly [p<0.05] lower than
control. [G1=5.0; G2=3.8; G3=4.1; G4=3.3; G5=4.0]. Day 14 survival
rates for G1 through G5 were 23%; 69%, 38%, 100% and 46%,
respectively. Thus, topical administration of IL-11 was shown to
favorably modify the course of chemotherapy-induced ulcerative
mucositis relative to control.
[0060] The present investigation demonstrates the beneficial
effects of a topical formulation of recombinant human IL-11
(rhIL-11) on the induction, course and resolution of
chemotherapy-induced mucositis.
[0061] Materials and Methods
[0062] Male LVG Golden Syrian hamsters (Charles River Laboratories,
Wilmington, Mass.), aged 5-6 weeks, were caged in small groups and
fed standard hamster chow and water ad libitum. All procedures were
conducted in accordance with guidelines set by the Harvard Medical
Area Standing Committee on Animals.
[0063] Mucositis induction and evaluation
[0064] All procedures were performed using anesthesia induced by
diethyl ether. To induce mucositis, 5-fluorouracil was administered
(60 mg/kg) by intraperitoneal injection on days 0 and 4. To mimic
the functional trauma, which occurs in humans, the left buccal
pouch mucosa was superficially irritated on day 4. Animals were
weighed and observed daily through day 14. Mucositis was evaluated
on days 6 to 14. To evaluate mucositis, animals were anesthetized
and the cheek pouch was everted, immobilized, and photographed
using a standard, validated technique. At the conclusion of the
study, all film was developed simultaneously. Photographs were
numbered, randomized and graded blindly by three independent
observers using a 10-point scale in which mild mucositis equaled a
score of 1 to 3, moderate 4 to 6, and severe greater than 6. Mild
mucositis was defined as mucosal erythema with vasodilation. Mold
mucositis was defined as ulceration with pseudomembrane formation,
erythema and vasodilation, and severe mucositis consisted of
diffuse ulceration with extensive areas of necrosis,
pseudomembranes, erythema and vasodilation.
[0065] Test groups and protocol
[0066] Ninety-five animals were prospectively randomized into five
equal groups. Each animal was individually numbered using an ear
punch technique. The following groups were studied:
2 Group 1 Vehicle control, topical, four times daily, days 3 to 14
(day 0 = day of first chemotherapy injection). Group 2 100 .mu.g
IL-11 per day in a divided dose given twice daily, subcutaneously,
days 0-14. Group 3 400 .mu.g IL-11 per day in a divided dose given
four times daily, topically, days 3-14. Group 4 100 .mu.g IL-11 per
day in a divided dose given twice daily, subcutaneously, days 3-14
Group 5 400 .mu.g Il-11 per day in a divided dose given four times
daily, topically, days 0-14.
[0067] Animals in the Group 1 were treated with topical application
of PBS containing 0.5% hamster serum. Animals in the test groups
received rhIL-11 made up in PBS containing 0.5% hamster serum.
Animals receiving topical therapy received treatment at 9 a.m.,
11:30 a.m., 2 p.m. and 4:30 p.m. Animals receiving injections of
IL-11 were treated at 9 a.m. and 4 p.m. Whole blood was obtained on
days 6 and 14 to evaluated the number of platelets.
[0068] Results
[0069] Cumulative mean mucositis scores for group 2 (3.0) and group
4 (2.7) were markedly less than that noted for the control (4.7).
Topical application of rhIL-11 was of benefit (group 3=3.3; group
5=3.4), though slightly less efficacious than rhIL-11 delivered by
the subcutaneous route. Interestingly, the efficacy of rhIL-11
given by either route was essentially comparable after day 10.
Marked rhIL-11 benefit was noted from day 6 through day 14 among
animals treated via the subcutaneous route. On days 6, 7, and 8,
animals in group 2 (IL-11 from day 0) had slightly less mucositis
than did animals in group 4 (IL-11 beginning on day 3). On day 9
this trend was reversed. No difference was seen for the remaining
days of the study.
[0070] Schedule of rhIL-11 administration had no observed effect on
its topical efficacy, with no significant differences observed
between scores of animals in groups 3 and 5. Less mucositis was
observed in animals treated with topically applied rhIL-11
throughout the course of the experiment. Reduction in mucositis
scores was essentially equivalent between both topical groups.
Interestingly, the effect of topically applied rhIL-11 was the same
as the subcutaneously administered peptide from day 11 to the
conclusion of the experiment.
[0071] Evaluation of the effect of rhIL-11 on moderate to severe
mucositis demonstrated a similar trend. From day 6 to day 9, the
percentage of control animals (group 1) with moderate to severe
mucositis was markedly greater than that noted in animals treated
with rhIL-11. In general, animals treated subcutaneously responded
more favorably. For example, on day 8, 67% of control animals had
mucositis scores equal to or greater than 5 compared to 13% of
animals in group 2 and 31% of animals in group 4. The frequency of
animals in the topically treated groups was 42% (group 3) and 39%
(group 5). This trend appears to have shifted on days 10 to 11
where it appears that topically treated animals did better than
those injected with rhIL-11. While this is true of surviving
animals, evaluation of survival data is of importance in
interpreting this apparent trend.
[0072] The above results demonstrate that topical administration of
rhIL-11 is beneficial in vivo for the treatment of oral
mucositis.
EXAMPLE 3
Treatment of Colitis (Colonic Ulcers)
[0073] The effects of three different dosages of recombinant human
interleukin-11 (rhIL-11), given subcutaneously (SC) either prior to
or subsequent to intracolonic administration of trinitrobenzene
sulfonic acid (TNB), were studied in Sprague-Dawley rats. The TNB
or control were given in a 40% ethanol solution to 312 anesthetized
adult male rats allotted to one of 26 groups (n=12). Control groups
were: subcutaneous (SC); saline alone; intrarectal (IR); 40%
ethanol alone; TNB alone; 40% ethanol alone, and SC, rhIL-11 at the
highest dosage alone and groups combining TNB with rhIL-11 therapy,
testing three dosages (100, 300, and 1,000 .mu.g/kg), given either
before or after induction of colitis with TNB. Body weight changes
were monitored. Rats were euthanized at 3 days, 7 days, or 14 days
after TNB administration. At necropsy, samples were collected to
evaluate fecal occult blood, mucosal myeloperoxidase activity and
mucosal gross indexes of ulceration. Histopathologic and
ultrastructural analyses of the colonic mucosa were performed. The
TNB alone elicited a prolonged, severe colitis in treated animals,
and the ethanol control group showed a short-lasting, less severe
colonic inflammatory response. Colonic ulcer indexes of rhIL-11
treated rats showed a consistent, dose-related reduction in the
severity of the TNB-induced colitis, whether the interleukin was
given before or after the TNB. This reduction was significant
(P<0.05) after administration of the intermediate (300 .mu.g/kg)
and highest (1,000 .mu.g/kg) dose levels of rhIL-11, in the groups
given rhIL-11 for 7 days after TNB. Myeloperoxidase activity was
increased during the TNB-induced colitis and was reduced by rhIL-11
administration (P<0.01). Fecal occult blood loss increased with
colitis and paralleled its severity. rhIL-11 enhanced mucus
production and decreased the severity of TNB-induced colitis.
EXAMPLE 4
Treatment of Mucositis
[0074] One of the dose-limiting toxicities of radiation therapy for
head and neck cancer is the occurrence of oral mucositis. The
effects of topical administration of IL-11 on this effect are
examined in conjunction with at least one of several regimens
employed for the treatment of advanced head and neck cancer. The
specifics of the regimens vary, and may include external beam
standard fractionated irradiation, hyperfractionated irradiation,
or accelerated fractionated irradiation. Various chemotherapy
agents may also be employed either sequentially or concurrently
with radiotherapy. Chemotherapy agents frequently employed in this
setting include 5-fluorouracil (5-FU), cisplatin, carboplatin,
hydroxyurea, paclitaxel, and topotecan, either alone or in various
combinations.
[0075] One regimen is outlined for the purpose of illustration. The
regimen involves hyperfractionated external beam irradiation and
concurrent chemotherapy with infusional 5-FU and cisplatin. This
regimen has been employed for several years for patients with
locally advanced squamous cell carcinoma of the head and neck.
Patients receive 125 centi-Gray (cG) twice per week of external
beam irradiation 5 days per week. The total planned dose is 7000
cG. During weeks 1 and 6 of radiotherapy, patients are treated with
5-FU and cisplatin. 5-FU is administered as a 5-day continuous
infusion of 600 mg/m/day while cisplatin is administered as a bolus
injection of 12 mg/m/day for 5 days. There is a 7-day scheduled
treatment break after 4000 cG to allow mucositis to subside. The
entire planned time for completion of the course of irradiation and
concurrent chemotherapy is 7 weeks.
[0076] Patients are randomly assigned in a ratio of 2:1 to receive
either topical IL-11 or placebo. Patients receiving IL-1 will
receive either 50 ug/L or 150 .mu.g/mL in 15 to 30 mL of Brioschi
(sodium bicarbonate). A 5 mg vial of IL-11 or excipient alone
[placebo] first will be resuspended with 1 mL water for injection
(WFI). When 30 mL of Brioschi is used, 300 .mu.L (for 50 ug/mL
dose) or 950 .mu.L (for the 150 ug/mL dose) containing either IL-11
or excipient alone will be removed from the vial and added to 30 mL
Brioschi. This final preparation, containing either IL-11 or
placebo excipient alone, is swished and swallowed twice daily for 6
to 8 weeks beginning the day before radiotherapy. All patients are
assessed to determine the effects of the topical preparation of
IL-11 on patients receiving chemotherapy and/or radiotherapy for
head and neck cancer. Among the endpoints summarized are:
[0077] Incidence of oral mucositis of Grade 3 or higher, as
measured by the modified National Cancer Institute Common Toxicity
Criteria;
[0078] Duration of oral mucositis of Grade 3 or higher. Duration is
measured as (1) the time from onset of Grade 3 or higher until
resolution to less than Grade 3 and
[0079] (2) the number of days of oral mucositis of Grade 3 or
higher;
[0080] Total duration of oral mucositis of any grade;
[0081] Maximum severity of oral mucositis experienced during the
study;
[0082] Incidence of hospitalization due to oral mucositis; and
[0083] Incidence of IV fluid resuscitation due to dehydration
secondary to poor oral intake [secondary to mucositis].
[0084] Patients receiving IL-11 exhibit significant improvement in
one or more of the above criteria.
EXAMPLE 5
Treatment of Psoriasis
[0085] The effects of IL-11 in modulating keratinocyte (the target
cell type in a number of cutaneous inflammatory skin diseases,
including psoriasis) growth and gene expression were assessed in
vitro. When rhIL-11 (recombinant human IL-11) was added to a basal
culture medium at a concentration of 100 ng/mL, it stimulated
keratinocyte proliferation. RT-PCR analysis showed that
keratinocytes grown in basal as well as enriched medium expressed
IL-11 receptor .alpha.-chain mRNA (IL-11R.alpha.). rhIL-11 was able
to stimulate phosphorylation of STAT-3 in keratinocytes when grown
in basal medium. To assess whether rhIL-11 could affect the levels
of surface intercellular adhesion molecule-1 (ICAM-1) induced by
IFN-.gamma. and TNF-.alpha., ketaninocytes were exposed to
IFN-.gamma. and TNF-.alpha. for one hour before adding rhIL-11.
Addition of rhIL-11 resulted in a significant decrease in the
amount of surface ICAM-1. These results show that IL-11 has direct
effects on keratinocytes, thereby downregulating cutaneous immune
responses.
[0086] While the present invention has been described in terms of
specific methods and compositions, it is understood that variations
and modifications will occur to those skilled in the art upon
consideration of the present invention.
[0087] Numerous modifications and variations in the invention as
described in the above illustrative examples are expected to occur
to those skilled in the art and, consequently, only such
limitations as appear in the appended claims should be placed
thereon. Accordingly, it is intended in the appended claims to
cover all such equivalent variations which come within the scope of
the invention as claimed.
[0088] The disclosure of all of the publications cited herein are
hereby incorporated by reference.
* * * * *