USOO8182814B2
`
`US 8,182,814 B2
`(10) Patent No.:
`(12) United States Patent
`
`Baca et al.
`(45) Date of Patent:
`May 22, 2012
`
`(54) METHODS OF TREATING INFLAMMATORY
`AIRWAY CONDITIONS BY INHIBITION OF
`IL-11 ACTIVITY
`
`(58) Field of Classification Search ........................ None
`See application file for complete search history.
`
`(75)
`
`Inventors: Manuel Baca, Gaithersburg, MD (US);
`Andrew Donald Nash, Kew (AU); Jack
`A. Elias, Woodbridge, CT (US)
`
`(73) Assignees: CSL Limited, Parkville, Vlctoria (AU);
`Yale University, New Haven, CT (US)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`
`(21) App1.No.: 12/256,883
`
`(22)
`
`Filed:
`
`Oct. 23, 2008
`
`(65)
`
`Prior Publication Data
`
`US 2009/0202533 A1
`
`Aug. 13, 2009
`
`Related U.S. Application Data
`
`(60) Provisional application No. 61/000,588, filed on Oct.
`26, 2007.
`
`(51)
`
`Int. Cl.
`(2006.01)
`A61K 38/20
`(2006.01)
`C07K 14/715
`(2006.01)
`C07K 14/54
`(2006.01)
`C07K 16/28
`(2006.01)
`C07K 16/24
`(52) U.S. Cl.
`............... 424/1431; 424/1331; 424/139.1;
`424/141.1; 424/145.1; 530/351; 530/387.3;
`530/388.22; 530/388.23
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`5,679,339 A * 10/1997 Keith et 31.
`.................. 424/852
`6,540,993 B1 *
`4/2003 W3rne et 31.
`..... 424/852
`
`7,252,820 B2 *
`8/2007 Boodhoo et 31.
`424/9467
`7,393,532 B1 *
`7/2008 de Sauv3ge et 31.
`....... 424/1441
`7,612,181 B2 * 11/2009 Wu et 31.
`.................... 530/387.3
`
`OTHER PUBLICATIONS
`
`Molet et 31., J. Allergy Clin. Immunology 2001, 108:430-438.*
`Wang, J. et 31. IL-11 selectively inhibits 3er0311ergen-induced pul-
`m0n3ry eosinophilia 3nd Th2 cytokine production. J Immunol 165,
`2222-31 (2000).*
`Minsh311 et 31., J. Allergy Clin. Immunology 2000, 105:232-238.*
`Kyung Sun Lee et 31., “Cysteinyl leukotriene upregulates IL-11
`expression in allergic airway disease of mice”, J. Allergy Clin.
`Immunol., 119(1): 141-149 (J3n. 1997; published online on Oct. 27,
`2006).
`
`* cited by examiner
`
`Primary Examiner 7 Lorraine Spector
`(74) Attorney, Agent, or Firm 7 Scu11y, Scott, Murphy &
`Presser, PC.
`
`(57)
`
`ABSTRACT
`
`The present invention provides a method for the treatment or
`prophylaxis of T—helper type 2 (Th2)-mediated disorders
`using antagonists of IL-1 1.
`
`16 Claims, 4 Drawing Sheets
`
`Lassen — Exhibit 1005, p. 1
`
`Lassen - Exhibit 1005, p. 1
`
`

`

`US. Patent
`
`May 22, 2012
`
`Sheet 1 of4
`
`US 8,182,814 B2
`
`24 HOURS
`
`D'O'OzO{OQA
`
`o 0
`
`v‘
`
`1'}A
`
`
`
`
`
`BALCellRecovery(x1004)
`
`(”NOD
`
`coo
`
`50.0
`
`40.0
`
`30.0
`
`20.0
`
`10.0
`
`0.0
`
`TOTALCELLS MACROPHAGE
`
`EOSINOPHIL
`
`LYMPHOCYTES NEUTROPHILS
`
`|L-11raKO+OVA
`
`DWT+SaIine
`
`.WT+OVA
`
`|L-11raKO+Sa|ine
`
`FIG. 1
`
`Lassen — Exhibit 1005, p. 2
`
`Lassen - Exhibit 1005, p. 2
`
`

`

`US. Patent
`
`May 22, 2012
`
`Sheet 2 of4
`
`US 8,182,814 B2
`
`48 HOURS
`
`" 70.0
`
`x1004V
`
`
`
`BALCellRecovery
`
`1 o :35:A.0‘
`
`'4o9‘
`
`A A
`
`60.0
`
`50.0
`
`40.0
`
`30.0
`
`20.0
`
`10.0
`
`0.0
`
`TOTAL CELLS
`
`MACROPHAG
`
`EOSINOPHIL
`
`LYMPHOCYTES
`
`NEUTROPHILS
`
`|L-11ra KO+OVA
`
`FIG. 2
`
`Lassen — Exhibit 1005, p. 3
`
`Lassen - Exhibit 1005, p. 3
`
`

`

`US. Patent
`
`May 22, 2012
`
`Sheet 3 of4
`
`US 8,182,814 B2
`
`72 HOURS
`
`90.0
`
`
`
`
`
`BALCellRecovery(x1004)
`
`A
`
`80.0
`
`0103\l
`
`coo
`
`40.0
`
`30.0
`
`20.0
`
`10.0
`
`.
`
`FIG. 3
`
`Lassen — Exhibit 1005, p. 4
`
`Lassen - Exhibit 1005, p. 4
`
`

`

`US. Patent
`
`May 22, 2012
`
`Sheet 4 of4
`
`US 8,182,814 B2
`
`24 Hours
`
`,
`
`4—.
`a1.
`5—
`EC
`
`3DU '
`
`5U
`
`BAL Inflammation
`
`lclsaline
`I CIDVA
`[II] Misaline
`MIDVA
`
`MN“GU'IUDO:
`
`Figure 4
`
`Lassen — Exhibit 1005, p. 5
`
`Lassen - Exhibit 1005, p. 5
`
`

`

`US 8,182,814 B2
`
`1
`METHODS OF TREATING INFLAMMATORY
`AIRWAY CONDITIONS BY INHIBITION OF
`IL-11 ACTIVITY
`
`APPLICATION DATA
`
`This application claims priority from and the benefit of
`US. Provisional Patent Application No. 61/000,588, filed
`Oct. 26, 2007, the entire contents of which are incorporated
`herein by reference.
`
`FIELD
`
`The present invention provides a method for the treatment
`or prophylaxis of T—helper type 2 (Th2)-mediated disorders
`using antagonists of lL-l 1.
`
`BACKGROUND
`
`Bibliographic details of references provided in the subject
`specification are listed at the end of the specification.
`Reference to any prior art is not, and shouldnot be taken as,
`an acknowledgment of or any form of suggestion that this
`prior art forms part of the common general knowledge in any
`country.
`Th2 cytokines, lL-4, lL-5, lL-9 and IL- 1 3, are derived from
`T helper type 2 (Th2) cells, although they may also derive
`from other cell types. These Th2 cytokines play an important
`role in the pathophysiology of allergic diseases including
`asthma.
`Asthma is a chronic disease that involves inflammation of
`
`the pulmonary airways and bronchial hyper-responsiveness
`leading to reversible obstruction of the lower airways (re-
`viewed in Bousquet et al, Am JRespir Crit Care Med 161(5):
`1720-1745, 2000). In a diagnostic context bronchial hyper-
`responsiveness is evidenced by decreased bronchial airflow
`following exposure to methacholine or histamine. Natural
`triggers that provoke airway obstruction include respiratory
`allergens, cold air, exercise, viral upper respiratory infection,
`and cigarette smoke. Bronchial provocation with allergen
`induces a prompt early phase immunoglobulin E (lgE)-me-
`diated decrease in bronchial airflow followed in many
`patients by a late-phase lgE-mediated reaction with a
`decrease in bronchial airflow for 4-8 hours.
`
`Asthmatic airways display lung hyperinflation, smooth
`muscle hypertrophy,
`fibrosis in the lamina reticularis,
`mucosal edema, epithelial cell sloughing, cilia cell disrup-
`tion, and mucus gland hypersecretion. Microscopically,
`asthma is characterized by the presence of increased numbers
`of eosinophils, mast cells, neutrophils,
`lymphocytes, and
`plasma cells in the bronchial tissues, bronchial secretions,
`and mucus. Activated CD4 T—lymphocytes that produce a Th2
`pattern of cytokines appear to be central to the initiation,
`development and maintenance of the disease phenotype
`(Robinson et al, NEnglJMed 326(5):298-304, 1992; Wills-
`Karp et al, Science 282(5397):2258-2261, 1998; Hamid et al
`J Clin Invest 87(5):1541-1546, 1991; Ray and Cohn, J Clin
`Invest 104(8):985-993, 1999). For example, the cytokines
`produced by these cells (including lL-4, lL-5, lL-9 and lL-13)
`regulate infiltration and mediator release by inflammatory
`cells and allergen specific antibody isotype switching from
`lgM to lgE. The activity of non-hemopoietic cells, for
`example mucus hypersecretion by goblet cells, is also regu-
`lated by Th2 cytokines.
`Regardless ofthe triggers of asthma, the repeated cycles of
`inflammation in the lungs with injury to the pulmonary tis-
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`sues followed by repair may produce long-term structural
`changes (“remodeling”) of the airways.
`In the most widely used animal model of human asthma,
`mice are sensitized to ovalbumin (ova, formulated in alum
`adjuvant) via the intraperitoneal route on one or more occa-
`sions.An allergic airway response is subsequently induced by
`single or repeated exposure to aerosol ova (generated via and
`ultrasonic nebulizer). Response parameters assessed over the
`subsequent 24-72 hr period include, for example, the accu-
`mulation of inflammatory cells and mediators in bronchoal-
`veolar lavage (BAL)
`fluid, bronchorestriction following
`intravenous administration of methacholine (airway hyper-
`reactivity) and ova specific serum lgE. Histological demon-
`stration ofinflammatory cell accumulation in lung tissues and
`goblet cell hyperplasia/metaplasia and associated mucus
`hypersecretion are also key characteristics of the mouse air-
`way response to ova. Large animal models of asthma (for
`example non-human primates and sheep), where lung archi-
`tecture, circulation and innervation more closely resemble
`that of humans, have been described but are less widely used.
`At the time studies described in this specification were per-
`formed there were no reports of the analysis of lL-l 1 antago-
`nists in either small or large animal models of asthma.
`lL-l 1 is a pleiotropic cytokine produced by a wide variety
`of cell types including fibroblasts, epithelial cells, chondro-
`cytes, endothelial cells, osteoblasts and certain tumor cells
`and cell lines (reviewed in Neben and Turner, Stem Cells.
`Suppl 2:156-62 1993, Du and Williams, Blood. 83(8):2023-
`2030, 1994). Human lL-11 is synthesized as a 19 kDa 199
`amino acid precursor protein, with a 21 amino acid leader
`sequence that is removed to generate a mature secreted pro-
`tein of 178 amino acids. lL-11 is highly conserved across
`speciesithe mature human and murine proteins share 88%
`homology at the amino acid level, while human and non-
`human primate lL-11 share 94% homology. Although the
`crystal structure of lL-11 has not been solved a variety of
`approaches (e.g. computer modeling and alanine scanning
`mutagenesis) suggest a 4 ot-helical bundle structure typical of
`many cytokines (Czupryn et al, Ann N YAcad Sci 762: 152-
`1 64, 1995).
`lL-11 was originally described as a soluble factor derived
`from stromal cells, which was capable of stimulating plasma-
`cytoma cell proliferation (Paul et al, Proc. Nat. Acad Sci.
`87:7512-7516, 1990). A variety of diverse biological proper-
`ties have subsequently been ascribed to lL-11 including: the
`ability to stimulate hemopoiesis, thrombopoiesis, megakary-
`opoiesis (Nandurkar et al, Blood 90:2148, 1997; Nakashima
`et al, Semin Hematol35(3):210-221, 1998), and bone resorp-
`tion (Sims et al, JBone Miner Res 20(7):1093-1102, 2005);
`the regulation of macrophage differentiation (Romas et al, J
`Exp Med 183(6):2581-2591, 1996); the regulation of proin-
`flammatory cytokine synthesis including TNFot and lL-lB
`(Leng etal,JImmunol159(5):2161-2168, 1997; Hermann et
`al, Arthritis Rheum 41 (8):1388-1397, 1998; Trepicchio et al,
`JImmunol 159(11):5661-5670, 1997); the ability to confer
`mucosal protection after chemotherapy and radiation therapy
`(Orazi et al, Lab Invest 75(1):33-42, 1996); and as an absolute
`requirement for normal development ofplacentation and sur-
`vival to birth (Robb et al, Nat Med 4:303, 1998). A number of
`these biological properties have been exploited in the devel-
`opment of new therapeutic strategies. Recombinant human
`lL-11 has been approved as a treatment for chemotherapy
`induced thrombocytopenia (Tepler et al, Blood 87(9):3607-
`3614, 1996) and is currently being assessed as a new approach
`to the treatment of chemotherapy induced gastrointestinal
`mucositis (Herrlinger et al, Am J Gastroenterol 101(4):793-
`797, 2006). Treatment with recombinant lL-11 in a mouse
`
`Lassen — Exhibit 1005, p. 6
`
`Lassen - Exhibit 1005, p. 6
`
`

`

`US 8,182,814 B2
`
`3
`model of rheumatoid arthritis (collagen induced arthritis,
`CIA) caused a significant reduction in the severity of estab-
`lished disease, which was associated with protection from
`joint damage, as assessed by histology (Walmsley et al,
`Immunology 95(1):31-37, 1998). In a subsequent Phase l/ll
`clinical study patients receiving a once weekly dose of lL-l 1
`(15 ug/kg) demonstrated a significant reduction in the number
`of tender joints, although there was no overall benefit at the
`ACR criterion of a 20% response (Moreland et al, Arthritis
`Res 3(4):247-252, 2001). Similarly, lL-11 has shown thera-
`peutic benefit in animal models of inflammatory bowel dis-
`ease (IBD; Peterson et al, Lab Invest 78(12): 1503-1512,
`1998) and this prompted clinical studies to assess the safety
`and efficacy of lL-11 in patients with active Crohns disease.
`While lL-11 was well tolerated and provided some clinical
`benefit, it remained significantly inferior when compared
`with a standard steroid based therapy (Herrlinger et al, supra
`2006).
`In addition to arthritis and lBD, lL-11 has also been dem-
`onstrated to provide therapeutic benefit in mouse (Lai et al,
`Nephron Exp Nephrol 101 (4):e146-1 54, 2005) and rat (Lai et
`al, JAm Soc Nephrol 12(11):2310-2320, 2001) models of
`glomerulonephritis. In these models, inflammatory disease is
`induced via the administration of ‘nephrotoxic serum’ (gen-
`erated by immunization of donor animals, for example sheep,
`with mouse or rat glomeruli preparations) and is assessed
`through standard histological and urine analysis.
`lL-11
`therapy resulted in a significant reduction in albuminuria at 24
`hrs as well as a decrease in fibrinogen deposition and infil-
`trating inflammatory cells at 14 days post induction ofdisease
`(Lai et al, supra 2005).
`In addition, lL-l 1 has been suggested as a potential thera-
`peutic agent in various other inflammatory disorders includ-
`ing radiation-induced lung damage (Redlich et al, JImmunol
`157(4):1i05 10, 1996), sepsis (Chang et al, Blood Cells Mol
`Dis 22(1):57-67, 1996) and psoriasis (Trepicchio et al, JClin
`Invest 104(11):1527-1537, 1999). U.S. Pat. No. 6,270,759
`suggests that lL-l 1 may be therapeutically useful for a variety
`of inflammatory conditions including asthma and rhinitis.
`The biological properties of lL-11 (IL-11 activity) are
`mediated through a multimeric receptor complex that incor-
`porates lL-11, the lL-llRot chain and gp130 (reviewed in
`Taga, JNeurochem 67(1): 1-10, 1996) and referred to as the
`lL-l 1 receptor complex. The lL-l 1R0. chain binds directly to
`lL-11 with low affinity (kDa ~10 nM), is unique to the lL-11
`receptor complex and is responsible for conferring specific-
`ity. gp130 is a shared receptor component used by members
`ofthe lL-6 ligand family (IL-6, lL-11, LlF, OSM and CNTF)
`and is responsible for the activation of intracellular signal
`transduction, primarily via the JAK/STAT pathway. Recent
`data suggests that the lL-l 1 receptor complex is a high aflinity
`(kDa ~400-800 pM), hexameric complex that incorporates
`two molecules of lL-l 1, two molecules of lL-l 1R0. and two
`molecules of gp130 (Barton et al, J Biol Chem 275(46):
`36197-36203, 2000).
`In contrast to the potential therapeutic approaches using
`lL-11, antagonists oflL-11 or lL-l 1R have been suggested as
`potential
`therapeutics for the treatment of osteoporosis
`(WO9959608) and in view of the role of lL-11 in the devel-
`opment ofplacentation and survival to birth (Robb et al, supra
`1998)
`as
`a
`contraceptive
`agents
`(WO9827996
`and
`W003099322).
`The role of lL-11 as a mediator of airway inflammation
`(including asthma) has primarily been investigated in mouse
`models, where one approach has been to assess the impact of
`increasing local
`lL-11 concentrations, Strategies used to
`achieve such an increase have included the local administra-
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`tion of recombinant lL-11 protein or local de novo synthesis
`via a lung specific lL-11 transgene. The results of these stud-
`ies have not been definitive and, in the context of airways
`disease such as asthma, the potential oflL-l 1 either as a target
`or as a novel therapeutic has remained unclear.
`Einarsson et al, JClin Invest 97(4):915-924, 1996 demon-
`strated that respiratory pathogens linked to asthma exacerba-
`tion (in contrast to other viral and bacterial pathogens) were
`potent stimulators of lung stromal cell lL-11 production in
`vitro. Consistent with this observation, lL-11 was readily
`detectable in aspirates from children with upper respiratory
`tract infections but not in aspirates from uninfected chil-
`dreniinterestingly the highest levels of lL-l 1 were detected
`in aspirates from children with clinical bronchospasm. When
`instilled into the lungs of mice, recombinant lL-11 induced a
`marked increase in sensitivity to methacholine and a mild
`mononuclear inflammatory response. In a subsequent report
`Tang et al, J. Clin. Invest. 98:2845, 1996 generated transgenic
`mice in which constitutive over-expression of lL-11 was tar-
`geted to the lung using the CC-10 promoter (CC-10/lL-11 Tg
`mice). In contrast to wildtype (wt) littermate controls, the
`transgenic animals demonstrated a nodular peribronchiolar
`mononuclear infiltrate with significant airways remodeling
`and sub-epithelial fibrosis. Furthermore, by two months of
`age the transgenic mice demonstrated increased airways
`resistance and airways hyperresponsivness to methacholine
`when compared with their wt littermates.
`While the above studies suggest that lL-11 overexpression
`may contribute to the development of experimental airways
`inflammation, a potential role for lL-11 in the pathology of
`asthma is less clear. For example, cell populations known to
`be central to the development of asthma pathology such as
`eosinophils and mast cells were not detected in the infiltrates
`induced by lL-l 1. Nevertheless lL-l 1 mRNA and protein has
`been detected in the epithelial and sub-epithelial layers of
`human bronchial biopsies, with levels significantly greater in
`moderate and severe asthmatics compared to patients with
`mild disease and non-asthmatics (Minshall et al, J Allergy
`Clin Immunol 105(2 Pt 1):232-238, 2000).
`To address this particular issue more directly Wang et al, J.
`Immunol. 165:2222, 2000 assessed the development of
`experimental asthma (OVA sensitization model)
`in the
`CC-10/lL-11 Tg mice. As expected OVA challenge of sensi-
`tized wt mice caused airway eosinophilic inflammation, Th2
`cell accumulation, and mucus hypersecretion with mucus
`metaplasia. Increased levels of endothelial cell VCAM-1,
`mucin (Muc) 5ac gene expression and bronchoalveolar lav-
`age and lung lL-4, lL-5, and lL-13 protein and mRNA were
`also noted. In contrast, OVA challenged CC10/lL-11 Tg mice
`that overexpressed lL-11 in the lung demonstrated lower
`levels of tissue and bronchoalveolar lavage inflammation,
`eosinophilia, and Th2 cell accumulation, and significantly
`lower levels of VCAM-1 and lL-4, lL-5, and lL-13 mRNA
`and protein. These studies demonstrate that lL-11 selectively
`inhibits many of the hallmarks of asthma pathology and
`prompted the authors to suggest that recombinant lL-11
`might be used as a treatment for Th2 mediated disorders such
`as asthma.
`
`More recent studies in the development of Th2 mediated
`disease have only served to add an additional layer of com-
`plexity (Chen et al, JImmunol 174(4):2305-2313, 2005). The
`Th2 cytokine lL-13 has been demonstrated to be key to the
`development of several aspects of asthma pathology includ-
`ing eosinophillic inflammation, mucus hypersecretion, air-
`ways hyper-responsiveness and allergen specific lgE. In
`agreement with these data lung-specific transgenic overex-
`pression of lL-13 (CC-10/lL-13 Tg mice) results in the devel-
`
`Lassen — Exhibit 1005, p. 7
`
`Lassen - Exhibit 1005, p. 7
`
`

`

`US 8,182,814 B2
`
`5
`opment of a severe Th2/asthma-like phenotype (Zhu et al, J
`Clin Invest 103(6):779-788, 1999). To assess a putative role
`for IL-1 1 in IL-13 activity (Chen et al, supra 2005) compared
`the expression of IL-1 1, IL-11R0t, and gp130 in lungs from
`wild-type mice and CC-10/IL-13 Tg mice and characterized
`the effects of a null mutation of IL-1 1R0. on the development
`of lung pathology in CC-10/IL-13 Tg mice. IL-13 was dem-
`onstrated to be a potent stimulator of IL-11 and IL-11R0t.
`Furthermore many ofthe pathological consequences of IL- 1 3
`overexpression, including inflammation, fibrosis, and mucus
`metaplasia, were substantially ameliorated in the absence of
`IL-1 1Rot. This led to the conclusion that IL-1 1R0. plays a key
`role in the pathogenesis of IL-13-induced inflammation and
`remodeling.
`Accordingly, with respect to airway-inflammation the role
`of IL-1 1 remains unclear. In contrast, for non-airway inflam-
`matory disease, the use of recombinant IL-11 as a novel
`therapeutic agent is well supported by published data.
`There is a need to develop new treatments for Th2-medi-
`ated disorders such as asthma.
`
`SUMMARY
`
`Throughout this specification, unless the context requires
`otherwise, the word “comprise”, or variations such as “com-
`prises” or “comprising”, will be understood to imply the
`inclusion of a stated element or integer or group of elements
`or integers but not the exclusion of any other element or
`integer or group of elements or integers.
`Nucleotide and amino acid sequences are referred to herein
`by a sequence identifier number (SEQ ID NO:). The SEQ ID
`NOs: correspond numerically to the sequence identifiers
`<400>1 (SEQ ID N021), <400>2 (SEQ ID NO:2), etc. A
`summary of the sequence identifiers is provided in Table 1.A
`sequence listing is provided after the claims.
`The present
`invention relates generally to the use of
`antagonists of IL-11 or IL-11Rot in the treatment of Th2-
`mediated disorders. Th2-mediated disorders include inflam-
`
`matory disorders such as asthma, chronic obstructive pulmo-
`nary disease (COPD), rhinitis, allergies and atopic dermatitis.
`Inparticular, the present invention provides the use of antago-
`nists of IL-11 or IL-11Rot in the treatment of asthma.
`
`The present invention is predicated in part on the elucida-
`tion of the role of IL-1 1 in an animal model of Th2-mediated
`
`inflammatory disorders such as asthma, and the effects of an
`antagonist of IL-11 or IL-11Rot in that model. In accordance
`with the present invention, inhibiting the activity of IL-11 is
`proposed to be useful
`in the treatment of Th2-mediated
`inflammatory disorders such as asthma, COPD, rhinitis, aller-
`gies and atopic dermatitis.
`Accordingly, one aspect ofthe present invention provides a
`method for the treatment of a Th2 -mediated disorder in a
`
`subject, the method comprising administering to the subject
`an amount of an antagonist of IL-1 1 or IL-1 1R0t. Reference to
`“an amount” includes an effective amount or an amount suf-
`
`ficient to ameliorate the symptoms of the Th2-mediated
`inflammatory disorder.
`In a particular embodiment, the Th2-mediated disorder is
`asthma.
`
`In another aspect the present invention provides a method
`for the treatment of asthma in a subject, the method compris-
`ing administering to the subject an amount of an antagonist of
`IL-11 or IL-11Rot.
`
`Particular antagonists include an IL-1 1 mutein, an anti-IL-
`1 1 antibody, an anti-IL-l 1R0. antibody and a soluble
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`IL-1 1R0. or functional part thereof. A “functional part” is that
`part of the antagonist that retains inhibitory activity towards
`IL-11 or IL-11Rot.
`
`Generally, the agent is administered in an amount and for a
`time and under conditions suflicient to ameliorate the symp-
`toms of the Th2-mediated inflammatory disorder.
`The administration may be systemic or local. Systemic
`administration is particularly useful. Reference to “systemic
`administration” includes intra-articular, intravenous, intrap-
`eritoneal, and subcutaneous injection, infusion, as well as
`administration via oral, rectal and nasal routes, or via inhala-
`tion. Administration by subcutaneous injection or via inhala-
`tion is particularly useful.
`The present invention further contemplates combination
`therapy such as targeting IL-11 and/or IL-11Rot and one or
`more other inflammatory targets.
`invention
`Accordingly, another aspect of the present
`relates to a method for the treatment of a Th2-mediated dis-
`
`order such as but not limited to asthma in a subject, the
`method comprising administering an antagonist of IL-11 or
`IL-11Rot and at least one other therapeutic agent such as an
`anti-inflammatory agent, a bronchodilator or an antibiotic.
`The co-administration may be simultaneous or sequential
`administration.
`
`Particular subjects are mammals such as humans.
`The present invention extends to the use of pharmaceutical
`compositions comprising antagonists of IL-11 or IL-11R0t.
`Useful compositions comprise an IL-11 mutein, an anti-IL-
`1 1 antibody, an anti-IL-l 1R0. antibody, or a soluble IL-1 1Rot.
`The present invention further provides the use ofan antago-
`nist of IL-1 1 or IL-1 1R0. in the manufacture of a medicament
`
`for the treatment of a Th2-mediated disorder in a subject.
`The present invention further provides the use ofan antago-
`nist of IL-1 1 or IL-1 1R0. in the manufacture of a medicament
`
`for the treatment of asthma in a subject.
`A medical kit is also provided comprising an antagonist of
`IL-11 or IL-11Rot
`together with instructions to use the
`antagonists in the treatment of a Th2 -mediated disorder such
`as asthma.
`
`A summary of sequence identifiers used throughout the
`subject specification is provided in Table 1.
`
`TABLE 1
`
`Summafl of sequence identifiers
`
`SEQUENCE
`ID NO:
`
`1
`2
`3
`4
`5
`6
`
`DESCRIPTION
`
`Murine IL- 11 mutein
`Murine IL- 11 mutein
`Murine IL- 11 mutein
`Human IL- 11 mutein
`Muc Sac primer for rtPCR
`Muc Sac primer for rtPCR
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`FIGS. 1, 2 and 3 are graphical representations showing the
`cell population in bronchoalveolar lavage (BAL) samples
`from wild type and IL-1 1R0. null mice challenged with phos-
`phate buffered saline (PBS) control and with OVA at 24, 48
`and 72 hours following exposure.
`FIG. 4 is a graphical representation showing the cell popu-
`lation in BAL samples from wild type mice challenged with
`OVA and treated with either an antagonist of IL-11/IL-1 1R0.
`or with a control.
`
`Lassen — Exhibit 1005, p. 8
`
`Lassen - Exhibit 1005, p. 8
`
`

`

`US 8,182,814 B2
`
`7
`DETAILED DESCRIPTION
`
`The singular forms “a”, “an” and “the” include plural
`aspects unless the context clearly dictates otherwise. Thus,
`for example, reference to “a cytokine” includes a single
`cytokine as well as two or more cytokines; reference to “an
`antibody” includes a single antibody, as well as two or more
`antibodies; reference to “the invention” includes single and
`multiple aspects, of an invention; and so forth.
`The present invention relates to a method for the treatment
`and prophylaxis of inflammatory conditions. It is predicated
`in part on an analysis of the use of a murine OVA-model of
`allergic asthma as a model of Th2-mediated inflammatory
`disorders.
`
`In this model, parameters of Th2 lung inflammation,
`mucus metaplasia and total and antigen-specific serum IgE
`levels enable the determination of the effectiveness of poten-
`tial therapeutic approaches in suppressing some of the key
`features of asthma.
`
`Infiltration of inflammatory cells into the airways, in par-
`ticular eosinophils, is an indicator of airway inflammation
`and a feature of asthmatic airways. The murine OVA-model
`of asthma results in a significant increase in the numbers of
`eosinophils and to a lesser extent macrophages migrating into
`the airways which can be easily seen in cell counts of fluid
`lavaged from the bronchoalveolar.
`In accordance with the present invention, inhibition of
`IL-11 activity including signaling with a test antagonist sig-
`nificantly impacts on the numbers of eosinophils and mac-
`rophages migrating into the airways as determined by cell
`counts of fluid lavaged from the bronchoalveolar of OVA-
`challenged mice, indicating that the antagonism of IL-11
`through inhibition of the formation of the IL-11 receptor
`complex is a useful therapeutic approach. These results were
`supported by experiments comparing IL-11Rot1 null mice
`with wildtype mice in the OVA-model of allergic asthma. The
`IL-11Rot1 null exhibited a reduction in the inflammatory
`response further suggesting that an antagonist of IL-11 or
`IL-11Rot a useful therapeutic approach.
`Accordingly, the present invention provides a method for
`the treatment of a Th2-mediated disorder in a subject, the
`method comprising administering to the subject an amount of
`an antagonist of IL-11 or IL-11R0t.
`Th2-mediated disorders include asthma, COPD, rhinitis,
`allergies and atopic dermatitis. A particular Th2-mediated
`disorder is asthma. Hence, another aspect the present inven-
`tion is directed to a method for the treatment of asthma in a
`subject, the method comprising administering to the subject
`an amount of an antagonist of IL-11 or IL-1 1R0t.
`Particular antagonists of IL-11 or IL-11Rot include an
`IL-1 1 mutein and an antibody specific for IL-1 1 or specific for
`IL-11Rot and a soluble IL-11Rot or a functional part thereof.
`Such a part is functional in the sense that it can still inhibit
`IL-1 1-mediated signaling.
`Reference to “amount” includes an effective amount or an
`
`amount suflicient to ameliorate symptoms of the Th2-medi-
`ated disorder.
`
`55
`
`The term “an antagonist of IL-11 or IL-11Rot” as used
`herein means an agent that binds to IL-11 or IL-11Rot and
`directly inhibits the formation on cells of a multimeric recep-
`tor complex that incorporates IL-11, IL-11Rot and gp130,
`thus inhibiting IL-11 signaling through the IL-11 receptor
`complex. Such antagonists inhibit the action of IL-11 on
`IL-11 sensitive cells. Examples of antagonists of IL-11 or
`IL-11Rot are:
`a. an IL-11 mutein;
`b. an antibody specific for IL-11;
`c. an antibody specific for IL-1 IR; and
`d. a soluble IL-11Rot.
`
`60
`
`65
`
`8
`Antagonists of IL-11 or IL-11Rot may also include agents
`that specifically inhibit expression of IL-11 or IL-11R, for
`example antisense polynucleotides that specifically recogn-
`ise a polynucleotide encoding IL-11 or the IL-11 receptor,
`interfering RNA that disrupt expression of IL-1 1 or the IL-1 1
`receptor or ribozymes that specifically recognise a polynucle-
`otide encoding IL-11 or the IL-11 receptor.
`An antibody specific for IL-11Rot and a soluble IL-11Rot
`may directly bind IL-11 and thereby directly inhibit the for-
`mation on cells of a multimeric receptor complex.
`Antagonists of IL-11 or IL-1 1R0. are known in the art, for
`example US. Pat. No. 6,998,123 describes a soluble
`IL-1 1Rot, IL-1 1-binding portions thereof, and commercially
`available antibodies to IL-11 and demonstrate their antago-
`nist activity. Soluble forms of IL-1 1R0. are also described in
`US. Pat. No. 6,528,281. International Patent Publication No.
`W0 03/099322 describes certain IL-11 muteins and demon-
`
`10
`
`15
`
`strates their antagonist activity.
`The term “IL-1 1” or its full name “interleukin-1 1” as used
`
`20
`
`25
`
`30
`
`35
`
`herein includes all mature forms of wild type mammalian
`IL-11, including murine, macaque and human, IL-1 1, and all
`truncated forms of such IL-11 that retain IL-11 signaling
`activity, i.e. the ability to bind with IL-11Rot and form a
`functional receptor complex with gp130. Mature human
`IL-11 is a 178 amino acid protein (i.e. lacking the 21 amino
`acid leader sequence of NPi000632, NCBI protein database
`Accession Number), and mature murine IL-1 1 is a 178 amino
`acid protein (i.e. lacking the 21 amino acid leader sequence of
`NPi032376, NCBI protein database Accession Number).
`The term “IL-1 1R0.” or its full name “interleukin-1 1 recep-
`tor alpha” as used herein includes, but is not limited to, human
`IL-11Rot having the nucleotide and amino acid sequences
`disclosed in SEQ ID NOs:1 and 2 of International Patent
`Publication NO. WO 96/19574 and murine IL-1 1R0. having
`the nucleotide and amino acid sequences disclosed in SEQ ID
`NOs:2 and 3 of International Patent Publication No. WO
`96/07737. IL-11Rot is also known as IL-11Rot1 and IL-11R
`
`40
`
`and the terms may be used herein interchangeably.
`The term “IL-1 1 mutein” as used herein refers to modified
`
`forms of mature IL-11 in which the amino acid sequence has
`been altered to retain effective binding to IL-1 1R0. but inhibit
`the formation of an IL-1 1 receptor complex with gp130. Such
`muteins compete with IL-11 for IL-11Rot binding and
`antagonize IL-11 signaling thereby inhibiting IL-11 action.
`Alterations to the sequence to form a mutein include amino
`acid substitutions of important residues for receptor binding.
`Conveniently, the mutein is based on human or murine IL-1 1
`and particularly human IL-11. International Publication No.
`WO 03/099322 describes certain IL-11 muteins and demon-
`
`45
`
`50
`
`strates their antagonist activity. Muteins may be expressed in
`suitable host cells and purified using standard techniques.
`IL-111 muteins may be further modified, for example to
`increase their in vivo half life, including for example, by the
`attachment of other elements such as a PEG groups. Methods
`for the PEGylation of peptides are well known in the art.
`The terms “antagonist”, “agent”,
`“compound”, and
`“active” may be used interchangeably herein to refer to a
`substance that induces a desired pharmacological and/or
`physiological effect and may include the IL-11 and IL-1 1R0.
`antagonists herein described. The terms also encompass phar-
`maceutically acceptable and pharmacologically active forms
`thereof, including salts. The desired effect is the inhibition of
`IL-11 activity or IL-11 receptor complex signaling.
`The terms “antibody” and “antibodies” include polyclonal
`and monoclonal antibodies and all the various forms derived
`
`from monoclonal antibodies, including but not limited to
`
`Lassen — Exhibit 1005, p. 9
`
`Lassen - Exhibit 1005, p. 9
`
`

`

`US 8,182,814 B2
`
`9
`full-length antibodies (e.g. having an intact Fc region), anti-
`gen-binding fragments, including for example, Fv, Fab, Fab'
`and F(ab')2 fragments; and antibody-derived polypeptides
`produced using recombinant methods such as single chain
`antibodies. The terms “antibody” and “antibodies” as used
`herein also refer to human antibodies produce