U.S. patent application number 12/322102 was filed with the patent office on 2009-11-05 for methods of treating inflammation using il-17 antagonists.
This patent application is currently assigned to Immunex Corporation. Invention is credited to Kendall M. Mohler.
Application Number | 20090274703 12/322102 |
Document ID | / |
Family ID | 22909803 |
Filed Date | 2009-11-05 |
United States Patent
Application |
20090274703 |
Kind Code |
A1 |
Mohler; Kendall M. |
November 5, 2009 |
Methods of treating inflammation using IL-17 antagonists
Abstract
Aspects of the present invention provide methods for treating
diseases associated with IL-17 mediated inflammatory or
immunoregulatory reactions.
Inventors: |
Mohler; Kendall M.;
(Poulsbo, WA) |
Correspondence
Address: |
IMMUNEX CORPORATION;LAW DEPARTMENT
1201 AMGEN COURT WEST
SEATTLE
WA
98119
US
|
Assignee: |
Immunex Corporation
Thousand Oaks
CA
|
Family ID: |
22909803 |
Appl. No.: |
12/322102 |
Filed: |
January 29, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12070981 |
Feb 22, 2008 |
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12322102 |
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10918084 |
Aug 13, 2004 |
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12070981 |
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10033522 |
Oct 18, 2001 |
6793919 |
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10918084 |
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60241230 |
Oct 18, 2000 |
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Current U.S.
Class: |
514/1.1 ;
424/141.1; 424/158.1 |
Current CPC
Class: |
A61K 2039/505 20130101;
A61K 38/1793 20130101; C07K 2319/30 20130101; A61P 19/02 20180101;
A61P 29/00 20180101; A61K 31/519 20130101; A61K 38/20 20130101;
C07K 2319/00 20130101 |
Class at
Publication: |
424/143.1 ;
424/158.1; 424/141.1; 514/2 |
International
Class: |
A61K 39/395 20060101
A61K039/395; A61K 38/00 20060101 A61K038/00 |
Claims
1. A method of treating an individual afflicted with a disease
associated with an IL-17-mediated inflammatory reaction, comprising
administering to the individual having a disease associated with an
IL-17-mediated inflammatory reaction an effective amount of an
IL-17 antagonist.
2. The method of claim 1, wherein the IL-17 antagonist is selected
from the group consisting of (a) a soluble IL-17 receptor; (b) an
antagonistic antibody that specifically binds IL-17; (c) an
antagonistic antibody that specifically binds IL-17 receptor; or
(d) combinations thereof.
3. A method of treating rheumatoid arthritis in an individual in
need thereof, comprising administering to the individual having
rheumatoid arthritis an effective amount of an IL-17
antagonist.
4. The method of claim 3, wherein the IL-17 antagonist is selected
from the group consisting of (a) a soluble IL-17 receptor; (b) an
antagonistic antibody that specifically binds IL-17; (c) an
antagonistic antibody that specifically binds IL-17 receptor; or
(d) combinations thereof.
5. The method of claim 2, wherein the antagonistic antibody that
specifically binds IL-17 is a monoclonal antibody.
6. The method of claim 4, wherein the antagonistic antibody that
specifically binds IL-17 is a monoclonal antibody.
7. The method according to claim 1, further comprising
administering, one or more therapeutics selected from the group
consisting of a TNF antagonist and an IL-1 antagonist.
8. The method of claim 7, wherein the IL-1 antagonist is selected
from the group consisting of soluble IL-1 receptor type II, IL-1 R
type I antibody, IL-1 receptor antagonist, and fusion protein
comprising soluble IL-1 receptor type I, and soluble IL-1 receptor
accessory protein.
9. The method of claim 7, wherein the TNF antagonist is selected
from the group consisting of TNF antibodies, soluble TNF receptor
p75, and soluble TNF receptor p55.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation of U.S. application Ser.
No. 12/070,981, filed Feb. 22, 2008, which is a continuation of
U.S. application Ser. No. 10/918,084, filed Aug. 13, 2004, now
abandoned, which is a divisional of U.S. application Ser. No.
10/033,522, filed Oct. 18, 2001, issued as U.S. Pat. No. 6,793,919
on Sep. 21, 2004, which claims the benefit under U.S.C. 119(e) of
U.S. provisional application Ser. No. 60/241,230, filed Oct. 18,
2000, all of which are incorporated by reference herein.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The invention pertains to methods for treating certain
diseases and disorders associated with inflammatory and
immunoregulatory responses. More particularly, the present
invention involves treating rheumatoid arthritis by administering
an IL-17 inhibitor or IL-17 antagonist, in particular IL-17
receptor, to an individual afflicted with such rheumatoid
arthritis.
[0004] 2. Description of Related Art
[0005] Cytokines are hormone-like molecules that regulate various
aspects of an immune or inflammatory response. Cytokines exert
their effects by specifically binding receptors present on cells,
and transducing a signal to the cells. Rouvier et al. (J. Immunol.
150:5445; 1993) reported a novel cDNA which they termed CTLA-8;
cloning of the human homolog led to the identification of this
family of molecules as Interleukin-17 (IL-17; Yao et al., Immunity
3:811; 1995). IL-17 is a cytokine produced by activated T cells
that stimulates the secretion of various proinflammatory molecules,
including tumor necrosis factor .alpha. (TNF-.alpha.),
Interleukin-1 .beta. (IL-1.beta.) and prostaglandin E.sub.2
(PGE.sub.2) from macrophages (Jovanovic et al., J. Immunol.
160:3513; 1998).
[0006] TNF-.alpha. and IL-1 are believed to play a role in the
inflammation and bone destruction that occurs in rheumatoid
arthritis (RA), albeit through different mechanisms (Joosten et
al., J. Immunol. 163:5049; 1999). Moreover, elevated levels of
IL-17 have been reported to occur in the synovial fluid of RA
patients, and may play a role in the bone destruction
characteristic of RA (Chabaud et al., Arthritis Rheum. 42:963,
1999; Jovanovic et al., Arthritis Rheum. 43:1134, 2000).
[0007] IL-17 acts on cells by binding to a specific receptor,
IL-17R, which was isolated as described U.S. Pat. No. 6,072,033,
issued Jun. 6, 2000. IL-17R is present on numerous cell types,
including synoviocytes and monocytes/macrophages. Although there
are numerous agents known in the art that are used in the treatment
of RA, there is a need to identify additional molecules that can be
used to treat or ameliorate the symptoms of this chronic
inflammatory disease.
SUMMARY OF THE INVENTION
[0008] The present invention relates to a method of treating a
mammal afflicted with a condition that relates to an inflammatory
response, in particular, rheumatoid arthritis, by administering an
IL-17 antagonist that inhibits IL-17 mediated signaling to a cell
via membrane-bound IL-17 receptor. Suitable IL-17 antagonists
include soluble IL-17 receptor, antagonistic antibodies that
specifically bind IL-17, antagonistic antibodies to IL-17 receptor
and combinations thereof.
[0009] Provided herein are methods for treating medical disorders
associated with IL-17 mediated inflammatory reactions or IL-17
mediated immunoregulatory reactions. The methods of the present
invention include administering an IL-17 antagonist, or IL-1
inhibitor, that inhibits IL-17 inflammatory or immunoregulatory
signaling, to an individual afflicted with an inflammatory or
immunoregulatory disease mediated by IL-17. More particularly, the
present invention involves administering an IL-17 antagonist such
as IL-17 receptor, to an individual inflicted with rheumatoid
arthritis, for a period of time sufficient to induce a sustained
improvement in the patient's condition.
DETAILED DESCRIPTION OF THE INVENTION
[0010] The present invention provides methods for treating an
individual including a human, who is suffering from a medical
disorder that is associated with IL-17 mediated inflammatory
reactions or IL-1 mediated immunoregulatory reactions. For purposes
of this disclosure, the terms "illness," "disease," "medical
condition" or "abnormal condition" are used interchangeably with
the term "medical disorder."
[0011] The subject methods involve administering to the patient an
IL-17 antagonist or IL-17 inhibitor that is capable of reducing the
effective amount of endogenous biologically active IL-17, by
preventing the binding of IL-17 to its receptor. Such antagonists
include receptor-binding peptide fragments of IL-17, antibodies
directed against IL-17 (antibodies that bind IL-17 and inhibit
binding thereof to IL-17 receptor), antibodies directed against
IL-17 receptor (antibodies that bind IL-17 receptor and inhibit
receptor binding of IL-17 without themselves transducing a signal
via IL-17 receptor), soluble forms of IL-17 receptor as discussed
herein, molecules that bind IL-17 or IL-17 receptor and inhibit the
interaction thereof and polypeptides comprising all or portions of
receptors for IL-17 or modified variants thereof, including
genetically-modified muteins, multimeric forms and
sustained-release formulations thereof. Particular antagonists are
soluble forms of IL-17 receptor. Other particular IL-17 antagonists
encompass chimeric proteins that include portions of both an
antibody molecule and an IL-17 antagonist molecule, particularly a
soluble portion of IL-17 receptor fused to an Fc. Such chimeric
molecules may form monomers, dimers or higher order multimers.
Preferred methods of the invention utilize IL-17 receptor in a form
that binds IL-17 and blocks IL-17 signal transduction, thereby
interrupting the proinflammatory and immunoregulatory effects of
IL-17.
[0012] The characterization, cloning and preparation of IL-17
receptor is described U.S. Pat. No. 6,072,033, issued Jun. 6, 2000,
incorporated herein by reference. The amino acid sequence of the
human IL-17 receptor (huIL-17 receptor) is shown in SEQ ID NO: 1.
The huIL-17 receptor has an N-terminal signal peptide with a
predicted cleavage site between amino acid 27 and 28. The signal
peptide is followed by a 293 amino acid extracellular domain, a 21
amino acid transmembrane domain, and a 525 amino acid cytoplasmic
tail. Soluble forms of huIL-17 receptor that are useful in the
methods of the present invention include the extracellular domain
(residues 1-320 of SEQ ID NO: 1 or residues 28-320 which excludes
the signal peptide) or a fragment of the extracellular domain that
has the properties of antagonizing or preventing binding of IL-17
receptor to IL-17. Other forms of the IL-17 receptor that are
useful in the present invention include muteins and variations that
are at least 70% or at least 90% homologous to the native IL-17
receptor of SEQ ID NO: 1 and as described in U.S. Pat. No.
6,072,033.
[0013] Other derivatives of the IL-17 receptor protein and homologs
thereof that are useful in the practice of this inventive method
include covalent or aggregative conjugates of the protein or its
fragments with other proteins or polypeptides, such as by synthesis
in recombinant culture as N-terminal or C-terminal fusions. For
example, the conjugated peptide may be a signal (or leader)
polypeptide sequence at the N-terminal region of the protein which
co-translationally or post-transitionally directs transfer of the
protein from its site of synthesis to its site of function inside
or outside of the cell membrane or wall (e.g., the yeast a-factor
leader).
[0014] Suitable forms of IL-17 inhibitors include chimeric proteins
which include a second polypeptide that may promote the spontaneous
formation by the chimeric protein of a dimer, trimer or higher
order multimer that is capable of binding IL-17 and preventing it
from binding to a cell-bound receptor that promotes IL-17 signaling
and inhibits or reduces the effects of inflammation and symptoms of
rheumatoid arthritis. Chimeric proteins used as antagonists may be
proteins that contain portions of an antibody molecule and a
soluble IL-17 receptor. Suitable fusion proteins include a IL-17
receptor polypeptide, e.g. the extracellular domain, or an IL-17
antagonistic fragment of the extracellular domain, linked to an
immunoglobulin Fc region. Fragments of an Fc region may also be
used, as well as Fc muteins that exhibit decreased affinity for Fc
receptors. A preferred Fc region is shown in SEQ ID NO:2. Depending
on the portion of the Fc region used, a fusion protein may be
expressed as a dimer, through formation of interchain disulfide
bonds. If the fusion proteins are made with both heavy and light
chains of an antibody, it is possible to form a protein oligomer
with as many as four IL-17 receptor regions.
[0015] Oligomeric forms of IL-17 inhibitors suitable for use in the
present invention also include an IL-17 receptor, the extracellular
domain of an IL-17 receptor, or an IL-17 inhibiting fragment of the
extracellular domain associated with a zipper domain, such as
zipper proteins described in U.S. Pat. No. 5,716,805, the
disclosure of which is incorporated by reference herein. Other
Examples of zipper domains are those found in the yeast
transcription factor GCN4 and a heat-stable DNA-binding protein
found in rat liver (C/EBP; Landschulz et al., Science 243:1681,
1989), the nuclear transforming proteins, fos and jun, which
preferentially form a heterodimer (O'Shea et al., Science 245:646,
1989; Turner and Tjian, Science 243:1689, 1989), and the gene
product of the murine proto-oncogene, c-myc (Landschulz et al.,
Science 240:1759, 1988). The fusogenic proteins of several
different viruses, including paramyxovirus, coronavirus, measles
virus and many retroviruses, also possess leucine zipper domains
(Buckland and Wild, Nature 338:547, 1989; Britton, Nature 353:394,
1991; Delwart and Mosialos, AIDS Research and Human Retroviruses
6:703, 1990). Examples of preferred zipper domains are those of SEQ
ID NO:3 and SEQ ID NO:4.
[0016] Other types of protein-based therapeutics are antibodies
that specifically recognize one or more epitopes of IL-17, or
epitopes of conserved variants of IL-17, or peptide fragments of
the IL-17 polypeptide that competitively inhibit IL-17 activity.
Antibodies to IL-17 can most conveniently be raised to a
recombinantly produced form of the protein. Or, antibodies that
specifically recognize a component of the IL-17 receptor and that
prevent signaling through the receptor by IL-17 can be used to
inhibit IL-17 activity. IL-17 antagonists that are antibodies
include but are not limited to polyclonal antibodies, monoclonal
antibodies (mAbs), humanized or chimeric antibodies, single chain
antibodies, Fab fragments, F(ab').sub.2 fragments, fragments
produced by a Fab expression library, anti-idiotypic (anti-Id)
antibodies, and epitope-binding fragments of any of the above.
Thus, such antibodies can, therefore, be utilized as part of
inflammatory disorder treatment methods.
[0017] For the production of antibodies, various host animals can
be immunized by injection with the IL-17 polypeptide, truncated
IL-17 polypeptides, a component of the IL-17 receptor (e.g., the
IL-17 extracellular region), a truncated version of a component of
the IL-17 receptor, and functional equivalents and mutants thereof.
Such host animals may include but are not limited to rabbits, mice,
and rats, to name but a few. Various adjuvants may be used to
increase the immunological response, depending on the host species,
including but not limited to Freund's (complete and incomplete),
mineral gels such as aluminum hydroxide, surface active substances
such as lysolecithin, pluronic polyols, polyanions, peptides, oil
emulsions, keyhole limpet hemocyanin, dinitrophenol, and
potentially useful human adjuvants such as BCG (bacille
Calmette-Guerin) and Corynebacterium parvum. Alternatively,
libraries of antibody fragments can be screened and used to develop
human antibodies through recombinant techniques. Such libraries are
commercially available from, for example, Cambridge Antibody
Technology (Melbourne, UK), and Morphosys (Munich, DE).
[0018] Monoclonal antibodies, which are homogeneous populations of
antibodies to a particular antigen, can be obtained by any
technique that provides for the production of antibody molecules by
continuous cell lines in culture. These include, but are not
limited to, the hybridoma technique of Kohler and Milstein, (U.S.
Pat. No. 4,376,110), the human B-cell hybridoma technique (Kosbor
et al., 1983, Immunology Today 4:72; Cole et al., 1983, Proc. Natl.
Acad. Sci. USA 80:2026-2030), and the EBV-hybridoma technique (Cole
et al., 1985, Monoclonal Antibodies And Cancer Therapy, Alan R.
Liss, Inc., pp. 77-96). Such antibodies may be of any
immunoglobulin class including IgG, IgM, IgE, IgA, IgD and any
subclass thereof. The hybridoma producing the mAb may be cultivated
in vitro or in vivo. Or, the antibody genes can be cloned and
optionally otherwise altered, and expressed in another cell line
approved for recombinant production of protein pharmaceuticals such
as, for example, CHO cells.
[0019] In addition, techniques developed for the production of
"chimeric antibodies" (Takeda et al., 1985, Nature, 314:452-454) by
splicing the genes from a mouse antibody molecule of appropriate
antigen specificity together with genes from a human antibody
molecule of appropriate biological activity can be used. A chimeric
antibody is a molecule in which different portions are derived from
different animal species, such as those having a variable region
derived from a porcine mAb and a human immunoglobulin constant
region.
[0020] Preferably, for use in humans, the antibodies are human or
humanized; techniques for creating such human or humanized
antibodies are also well known and are commercially available from,
for example, Protein Design Labs, Inc. (Fremont, Calif.), Medarex
Inc. (Princeton, N.J.) and Abgennix Inc. (Fremont, Calif.).
[0021] Techniques described for the production of single chain
antibodies (U.S. Pat. No. 4,946,778; Bird, 1988, Science
242:423-426; Huston et al., 1988, Proc. Natl. Acad. Sci. USA
85:5879-5883; and Ward et al., 1989, Nature 334:544-546) can also
be adapted to produce single chain antibodies against IL-17 gene
products and IL-17 receptor gene products. Single chain antibodies
are formed by linking the heavy and light chain fragments of the Fv
region via an amino acid bridge, resulting in a single chain
polypeptide.
[0022] Antibody fragments that recognize specific epitopes can be
generated by known techniques. For example, such fragments include
but are not limited to: the F(ab').sub.2 fragments which can be
produced by pepsin digestion of the antibody molecule and the Fab
fragments which can be generated by reducing the disulfide bridges
of the (ab').sub.2 fragments. Alternatively, Fab expression
libraries can be constructed (Huse et al., 1989, Science,
246:1275-1281) to allow rapid and easy identification of monoclonal
Fab fragments with the desired sThis invention additionally
provides for the use of soluble forms of IL-17 receptor, including
the extracellular domain and suitable fragments thereof in the
manufacture of a medicament for the prevention or treatment of
rheumatoid arthritis. This invention additionally provides for the
use of DNA encoding human IL-17 receptor, as described in U.S. Pat.
No. 6,072,033, in the manufacture of soluble IL-17 receptor for use
in the manufacture of a medicament for the treatment of rheumatoid
arthritis.
[0023] In one preferred embodiment of the invention,
sustained-release forms of soluble IL-17 receptor, or other IL-17
inhibitors described herein, are used. Sustained-release forms
suitable for use in the disclosed methods include, but are not
limited to, IL-17 receptor or other IL-17 inhibitor, that is
encapsulated in a slowly-dissolving biocompatible polymer, admixed
with such a polymer, and or encased in a biocompatible
semi-permeable implant. In addition, the soluble IL-17 receptor or
may be conjugated with polyethylene glycol (pegylated) to prolong
its serum half-life or to enhance protein delivery. Soluble forms
of IL-17 receptor, including monomers, fusion proteins (also called
"chimeric proteins), dimers, trimers or higher order multimers, are
useful in formulating IL-17 antagonists for treating rheumatoid
arthritis. Similarly, antibodies that antagonize the IL-17/IL-17R
interaction and signaling pathway are useful antagonists for
treating rheumatoid arthritis.
[0024] To treat rheumatoid arthritis, a molecule comprising an
IL-17 binding soluble IL-17 receptor, or antibody as described
herein, is administered to the patient in an amount and for a time
sufficient to induce a sustained improvement in at least one
indicator that reflects the severity of the rheumatoid arthritis.
An improvement is considered "sustained" if the patient exhibits,
or experiences as self-assessed, the improvement on at least two
occasions separated by one to four weeks. The degree of improvement
is determined based on signs or symptoms, and may also employ
questionnaires that are administered to the patient, such as
quality-of-life questionnaires.
[0025] Various indicators that reflect the extent of the patient's
rheumatoid arthritis may be assessed for determining whether the
amount and time of the treatment is sufficient. The baseline value
for the chosen indicator or indicators is established by
examination of the patient, or the patient's own self assessment,
prior to administration of the first dose of the soluble IL-17
receptor, or antagonistic antibody, as described above. Preferably,
the baseline examination is done within about 60 days of
administering the first dose.
[0026] Improvement is induced by repeatedly administering a dose of
soluble IL-17 receptor or other suitable IL-17 receptor derivative,
or antibody, as described herein, until the patient manifests an
improvement over baseline for the chosen indicator or indicators.
The degree of improvement is obtained by repeatedly administering
the medicament over a period of at least a month or more, e.g., for
one, two, or three months or longer, or indefinitely.
[0027] Any efficacious route of administration may be used to
therapeutically administer IL-17 receptor or antibody, as described
herein. If injected, a IL-17 inhibitor can be administered, for
example, via intra-articular, intravenous, intramuscular,
intralesional, intraperitoneal or subcutaneous routes by bolus
injection or by continuous infusion. Other suitable means of
administration include sustained release from implants, aerosol
inhalation, eyedrops, oral preparations, including pills, syrups,
lozenges or chewing gum, and topical preparations such as lotions,
gels, sprays, ointments or other suitable techniques.
Administration by inhalation is particularly beneficial when
treating diseases associated with pulmonary disorders.
Alternatively, IL-17 inhibitor polypeptides, such as a soluble
IL-17 receptor, may be administered by implanting cultured cells
that express the protein; for example, by implanting cells that
express a soluble IL-17 receptor. In one embodiment, the patient's
own cells are induced to produce by transfection in vivo or ex vivo
with a DNA that encodes an IL-17 inhibitor, and particularly
soluble IL-17 receptor. This DNA can be introduced into the
patient's cells, for example, by injecting naked DNA or
liposome-encapsulated DNA that encodes soluble IL-17 receptor, or
by other means of transfection. When soluble IL-17 receptor is
administered in combination with one or more other biologically
active compounds, these may be administered by the same or by
different routes, and may be administered simultaneously,
separately or sequentially.
[0028] Soluble IL-17 receptor or other antagonists of IL-17
preferably are administered in the form of a physiologically
acceptable composition comprising purified recombinant protein in
conjunction with physiologically acceptable carriers, excipients or
diluents. Such carriers are nontoxic to recipients at the dosages
and concentrations employed. Ordinarily, preparing such
compositions entails combining the IL-17 antagonist with buffers,
antioxidants such as ascorbic acid, low molecular weight
polypeptides (such as those having fewer than 10 amino acids),
proteins, amino acids, carbohydrates such as glucose, sucrose or
dextrins, chelating agents such as EDTA, glutathione and other
stabilizers and excipients. Neutral buffered saline or saline mixed
with conspecific serum albumin are exemplary appropriate diluents.
The IL-17 receptor or antagonistic antibody, preferably is
formulated as a lyophilizate using appropriate excipient solutions
(e.g., sucrose) as diluents. Appropriate dosages can be determined
in standard dosing trials, and may vary according to the chosen
route of administration. In accordance with appropriate industry
standards, preservatives may also be added, such as benzyl alcohol.
The amount and frequency of administration will depend, of course,
on such factors as the nature and severity of the indication being
treated, the desired response, the age and condition of the
patient, and so forth.
[0029] In one embodiment of the invention, the IL-17 inhibitor is
administered one time per week to treat rheumatoid arthritis, in
another embodiment is administered at least two times per week, and
in another embodiment is administered at least once per day. An
adult patient is a person who is 18 years of age or older. If
injected, the effective amount, per adult dose, ranges from 1-200
mg/m.sup.2, or from 1-40 mg/m.sup.2 or about 5-25 mg/m.sup.2.
Alternatively, a flat dose may be administered, whose amount may
range from 2-400 mg/dose, 2-100 mg/dose or from about 10-80
mg/dose. If the dose is to be administered more than one time per
week, an exemplary dose range is the same as the foregoing
described dose ranges or lower. Preferably, the IL-17 inhibitor is
administered two or more times per week at a per dose range of
25-100 mg/dose. In one embodiment of the invention, the various
indications described below are treated by administering a
preparation acceptable for injection containing IL-17 inhibitor at
80-100 mg/dose, or alternatively, containing 80 mg per dose. The
dose is administered repeatedly. If a route of administration other
than injection is used, the dose is appropriately adjusted in
accord with standard medical practices. For example, if the route
of administration is inhalation, dosing may be one to seven times
per week at dose ranges from 10 mg/dose to 50 mg per dose.
[0030] In many instances, an improvement in a patient's condition
will be obtained by injecting a dose of up to about 100 mg of IL-17
inhibitor one to three times per week over a period of at least
three weeks, though treatment for longer periods may be necessary
to induce the desired degree of improvement.
[0031] For pediatric patients (age 4-17), a suitable regimen
involves the subcutaneous injection of 0.4 mg/kg to 5 mg/kg of
IL-17 inhibitor, administered by subcutaneous injection one or more
times per week.
[0032] The invention further includes the administration of an
IL-17 inhibitor concurrently with one or more other drugs that are
administered to the same patient, each drug being administered
according to a regimen suitable for that medicament. This
encompasses pre-treatment, simultaneous treatment, sequential
treatment and alternating regimens. Examples of such drugs include
but are not limited to analgesics, corticosteroids, antagonists of
inflammatory cytokines, DMARDs, including methotrexate, and
non-steroidal anti-inflammatories. Additionally, IL-17 inhibitors
described herein, may be combined with each other or combined with
other molecules that reduce endogenous IL-17 levels.
[0033] In one preferred embodiment of the invention, methods for
treating rheumatoid arthritis include administering soluble IL-17
receptor or other IL-17 inhibitor described herein, in combination
with one or more additional cytokines or cytokine inhibitors. For
example, an IL-17 inhibitor may be administered in a composition
with agents that inhibit the interaction of inflammatory cytokines
with their receptors. Suitable agents that may be utilized in
combination with IL-17 inhibitors, but are not limited to, IL-1
inhibitors, such as type II IL-1 receptor, including IL-1 binding
fragments of type II IL-1 receptor, disclosed in U.S. Pat. No.
5,350,683; IL-1 binding and IL-1 inhibitory fragments of type I
IL-1 receptor; IL-1 receptor antagonist, IL-1 beta converting
enzyme (ICE) inhibitors, antibodies to IL-1, including IL-1 alpha
and IL-1 beta and other IL-1 family members, and therapeutics known
as IL-1 traps and antagonistic type I IL-1 receptor antibodies; TNF
inhibitors such as antagonistic TNF antibodies; soluble TNF
receptors p55 and p75, particularly ENBREL; IL-18 inhibitors
including IL-18 binding protein disclosed in WO 0012555; inhibitory
forms of IL-18 receptors, disclosed in WO 99/37772; antagonist
IL-18 antibodies, and antagonistic IL-18 receptor antibodies;
CD30-ligand inhibitors; and, CD4 inhibitors.
[0034] Specific IL-1 inhibitors include forms of IL-Ira described
in U.S. Pat. No. 5,075,222 and modified forms and variants
including those described in U.S. Pat. No. 5,922,573, WO 91/17184,
WO 92 16221, and WO 96 09323, all of which are incorporated herein
by reference. IL-1 beta converting enzyme (ICE) inhibitors include
peptidyl and small molecule ICE inhibitors including those
described in PCT patent applications WO 91/15577; WO 93/05071; WO
93/09135; WO 93/14777 and WO 93/16710; and European patent
application 0 547 699. Non-peptidyl compounds include those
described in PCT patent application WO 95/26958, U.S. Pat. No.
5,552,400, U.S. Pat. No. 6,121,266, Dolle et al., J. Med. Chem.,
39, pp. 2438-2440 (1996). Additional ICE inhibitors are described
in U.S. Pat. Nos. 6,162,790, 6,204,261, 6,136,787, 6,103,711,
6,025,147, 6,008,217, 5,973,111, 5,874,424, 5,847,135, 5,843,904,
5,756,466, 5,656,627, 5,716,929.
[0035] Further, suitable IL-1 antagonists encompass chimeric
proteins that include portions of both an antibody molecule and an
IL-1 antagonist molecule. Such chimeric molecules may form
monomers, dimers or higher order multimers. Other suitable IL-1
antagonists include peptides derived from IL-1 that are capable of
binding competitively to the IL-1 signaling receptor, IL-1 R type
I.
[0036] Additional inhibitors used in combination with IL-17
receptor include those that antagonize TGF.beta., IFN.gamma., IL-6
or IL-8. The cytokine inhibitors may be administered as separate
compositions, or together with IL-17 receptor, and the cytokine
inhibitors may be administered by the same or different routes.
[0037] Where the compounds are used together with one or more other
components, the compound and the one or more other components may
be administered simultaneously, separately or sequentially (usually
in pharmaceutical format).
[0038] It is understood that the response by individual patients to
the aforementioned medications or combination therapies may vary,
and the most efficacious combination of drugs for each patient will
be determined by the treating physician or physicians.
[0039] The following examples are offered by way of illustration,
and not by way of limitation. Those skilled in the art will
recognize that variations of the invention embodied in the examples
can be made, especially in light of the teachings of the various
references cited herein, the disclosures of which are incorporated
by reference herein.
Example 1
[0040] This example describes a construct for expression of an
IL-17R/Fc fusion protein. Construction of the IL-17 receptor DNA is
described in U.S. Pat. No. 6,072,033, issued Jun. 6, 2000. Briefly,
a soluble form of IL-17 receptor fused to the Fc region of human
IgG1 was constructed in the mammalian expression vector pDC409 by
utilizing a 980 bp DNA fragment (nucleotides encoding the amino
acid sequence of residues 1 to 322 of IL-17 receptor as shown in
SEQ ID NO:1) amplified from IL-17 receptor cDNA in a three way
ligation with a DNA fragment encoding human IgG1 Fc (SEQ ID NO:3)
and the plasmid pDC409 (described U.S. Ser. No. 08/235,397).
[0041] The IL-17 receptor/Fc expression plasmids were transfected
into mammalian cells (for example, CV-1/EBNA cells), and
supernatants collected. Following the collection, the IL-17
receptor/Fc fusion proteins were purified on a protein A sepharose
column (Pharmacia, Uppsala, Sweden) as described below. Protein
concentration was determined by an enzyme-linked immunoabsorbent
assay specific for the IgG Fc domain and by BCA analysis
(Pharmacia); purity was confirmed by SDS-polyacrylamide gel
electrophoresis analysis followed by silver stain of the gel.
Example 2
[0042] This example describes purification of IL-17 receptor fusion
proteins. IL-17 receptor/Fc fusion protein was purified by
conventional methods using Protein A or Protein G chromatography.
Approximately one liter of culture supernatant containing IL-17
receptor/Fc fusion protein was purified by filtering mammalian cell
supernatants (e.g., in a 0.45 m filter) and applying filtrate to a
protein A/G antibody affinity column (Schleicher and Schuell,
Keene, N.H.) at 4.degree. C. at a flow rate of 80 ml/hr for a 1.5
cm.times.12.0 cm column. The column was washed with 0.5 M NaCl in
PBS until free protein was not detected in the wash buffer.
Finally, the column was washed with PBS. Bound fusion protein was
eluted from the column with 25 mM citrate buffer, pH 2.8, and
brought to pH 7 with 500 mM Hepes buffer, pH 9.1.
Example 3
[0043] This example describes results obtained using IL-17 receptor
in a murine model of rheumatoid arthritis. Mice (male DBA/1 mice
five to eight weeks old) were immunized intradermally at the base
of the tail with 100 .mu.g type II collagen (CII) in complete
Freund's adjuvant (CFA). Twenty-one days later, the mice were
boosted with 200 .mu.g CII in incomplete Freund's adjuvant (IFA)
intradermally at the base of the tail. Signs of clinical arthritis
begin to appear in the mice three to five days after the
booster.
[0044] Mice were evaluated for signs of clinical score and disease
incidence three times weekly, beginning at the time of the booster.
Disease severity was evaluated using an established arthritis index
system. Each paw was assigned a clinical score based on the index.
Paw scores for each animal were combined to determine a total
cumulative score. The arthritis index used was: 0=normal
appearance; 1=erythema/edema in 1-2 digits; 2=erythema/edema in
more than two digits, or mild swelling in ankle/wrist joint;
3=erythema/edema in entire paw; 4=massive erythema/edema of entire
paw extending into proximal joints, ankylosis, loss of
function.
[0045] At the time of the booster, mice (15-20 mice per group) were
injected intraperitoneally with either 150 .mu.g rat IgG, 1 .mu.g
TNF receptor/Fc, 150 .mu.g IL-17 receptor/Fc as prepared in
Examples 1 and 2, or a combination of 1 .mu.g TNF receptor/Fc and
150 .mu.g IL-17 receptor/Fc. The treatment regimen was repeated
daily for fourteen days. The mice were evaluated for clinical score
and disease incidence three times weekly. The average final score
for each group is shown in Table 1.
TABLE-US-00001 TABLE 1 Decrease of Arthritis Symptoms in Mice Given
TNF receptor/Fc and/or IL-17 receptor/Fc Group: Treatment: Average
Final Score: Group 1 Rat IgG 8.4 Group 2 TNF receptor/Fc 5.7 Group
3 IL-17 receptor/Fc 5.1 Group 4 TNF receptor/Fc plus 1.7 IL-17
receptor/Fc
[0046] A second set of experiment using substantially the same
parameters was carried out. The average final score for each group
is shown in Table 2:
TABLE-US-00002 TABLE 2 Decrease of Arthritis Symptoms in Mice Given
TNF receptor/Fc and IL-17 receptor/Fc, Group: Treatment: Average
Final Score: Group 1 Rat IgG 9.2 Group 2 TNF receptor/Fc 5.9 Group
3 IL-17 receptor/Fc 3.9 Group 4 TNF receptor/Fc plus 5.0 IL-17
receptor/Fc
[0047] These results indicate that IL-17 receptor ameliorates the
symptoms of arthritis in an animal model of rheumatoid arthritis.
Moreover, IL-17 receptor may be used in combination with TNF
receptor (or other inhibitors of inflammation) to reduce the
severity of clinical arthritis.
Sequence CWU 1
1
41866PRTHomo sapiens 1Met Gly Ala Ala Arg Ser Pro Pro Ser Ala Val
Pro Gly Pro Leu Leu1 5 10 15Gly Leu Leu Leu Leu Leu Leu Gly Val Leu
Ala Pro Gly Gly Ala Ser 20 25 30Leu Arg Leu Leu Asp His Arg Ala Leu
Val Cys Ser Gln Pro Gly Leu 35 40 45Asn Cys Thr Val Lys Asn Ser Thr
Cys Leu Asp Asp Ser Trp Ile His 50 55 60Pro Arg Asn Leu Thr Pro Ser
Ser Pro Lys Asp Leu Gln Ile Gln Leu65 70 75 80His Phe Ala His Thr
Gln Gln Gly Asp Leu Phe Pro Val Ala His Ile 85 90 95Glu Trp Thr Leu
Gln Thr Asp Ala Ser Ile Leu Tyr Leu Glu Gly Ala 100 105 110Glu Leu
Ser Val Leu Gln Leu Asn Thr Asn Glu Arg Leu Cys Val Arg 115 120
125Phe Glu Phe Leu Ser Lys Leu Arg His His His Arg Arg Trp Arg Phe
130 135 140Thr Phe Ser His Phe Val Val Asp Pro Asp Gln Glu Tyr Glu
Val Thr145 150 155 160Val His His Leu Pro Lys Pro Ile Pro Asp Gly
Asp Pro Asn His Gln 165 170 175Ser Lys Asn Phe Leu Val Pro Asp Cys
Glu His Ala Arg Met Lys Val 180 185 190Thr Thr Pro Cys Met Ser Ser
Gly Ser Leu Trp Asp Pro Asn Ile Thr 195 200 205Val Glu Thr Leu Glu
Ala His Gln Leu Arg Val Ser Phe Thr Leu Trp 210 215 220Asn Glu Ser
Thr His Tyr Gln Ile Leu Leu Thr Ser Phe Pro His Met225 230 235
240Glu Asn His Ser Cys Phe Glu His Met His His Ile Pro Ala Pro Arg
245 250 255Pro Glu Glu Phe His Gln Arg Ser Asn Val Thr Leu Thr Leu
Arg Asn 260 265 270Leu Lys Gly Cys Cys Arg His Gln Val Gln Ile Gln
Pro Phe Phe Ser 275 280 285Ser Cys Leu Asn Asp Cys Leu Arg His Ser
Ala Thr Val Ser Cys Pro 290 295 300Glu Met Pro Asp Thr Pro Glu Pro
Ile Pro Asp Tyr Met Pro Leu Trp305 310 315 320Val Tyr Trp Phe Ile
Thr Gly Ile Ser Ile Leu Leu Val Gly Ser Val 325 330 335Ile Leu Leu
Ile Val Cys Met Thr Trp Arg Leu Ala Gly Pro Gly Ser 340 345 350Glu
Lys Tyr Ser Asp Asp Thr Lys Tyr Thr Asp Gly Leu Pro Ala Ala 355 360
365Asp Leu Ile Pro Pro Pro Leu Lys Pro Arg Lys Val Trp Ile Ile Tyr
370 375 380Ser Ala Asp His Pro Leu Tyr Val Asp Val Val Leu Lys Phe
Ala Gln385 390 395 400Phe Leu Leu Thr Ala Cys Gly Thr Glu Val Ala
Leu Asp Leu Leu Glu 405 410 415Glu Gln Ala Ile Ser Glu Ala Gly Val
Met Thr Trp Val Gly Arg Gln 420 425 430Lys Gln Glu Met Val Glu Ser
Asn Ser Lys Ile Ile Val Leu Cys Ser 435 440 445Arg Gly Thr Arg Ala
Lys Trp Gln Ala Leu Leu Gly Arg Gly Ala Pro 450 455 460Val Arg Leu
Arg Cys Asp His Gly Lys Pro Val Gly Asp Leu Phe Thr465 470 475
480Ala Ala Met Asn Met Ile Leu Pro Asp Phe Lys Arg Pro Ala Cys Phe
485 490 495Gly Thr Tyr Val Val Cys Tyr Phe Ser Glu Val Ser Cys Asp
Gly Asp 500 505 510Val Pro Asp Leu Phe Gly Ala Ala Pro Arg Tyr Pro
Leu Met Asp Arg 515 520 525Phe Glu Glu Val Tyr Phe Arg Ile Gln Asp
Leu Glu Met Phe Gln Pro 530 535 540Gly Arg Met His Arg Val Gly Glu
Leu Ser Gly Asp Asn Tyr Leu Arg545 550 555 560Ser Pro Gly Gly Arg
Gln Leu Arg Ala Ala Leu Asp Arg Phe Arg Asp 565 570 575Trp Gln Val
Arg Cys Pro Asp Trp Phe Glu Cys Glu Asn Leu Tyr Ser 580 585 590Ala
Asp Asp Gln Asp Ala Pro Ser Leu Asp Glu Glu Val Phe Glu Glu 595 600
605Pro Leu Leu Pro Pro Gly Thr Gly Ile Val Lys Arg Ala Pro Leu Val
610 615 620Arg Glu Pro Gly Ser Gln Ala Cys Leu Ala Ile Asp Pro Leu
Val Gly625 630 635 640Glu Glu Gly Gly Ala Ala Val Ala Lys Leu Glu
Pro His Leu Gln Pro 645 650 655Arg Gly Gln Pro Ala Pro Gln Pro Leu
His Thr Leu Val Leu Ala Ala 660 665 670Glu Glu Gly Ala Leu Val Ala
Ala Val Glu Pro Gly Pro Leu Ala Asp 675 680 685Gly Ala Ala Val Arg
Leu Ala Leu Ala Gly Glu Gly Glu Ala Cys Pro 690 695 700Leu Leu Gly
Ser Pro Gly Ala Gly Arg Asn Ser Val Leu Phe Leu Pro705 710 715
720Val Asp Pro Glu Asp Ser Pro Leu Gly Ser Ser Thr Pro Met Ala Ser
725 730 735Pro Asp Leu Leu Pro Glu Asp Val Arg Glu His Leu Glu Gly
Leu Met 740 745 750Leu Ser Leu Phe Glu Gln Ser Leu Ser Cys Gln Ala
Gln Gly Gly Cys 755 760 765Ser Arg Pro Ala Met Val Leu Thr Asp Pro
His Thr Pro Tyr Glu Glu 770 775 780Glu Gln Arg Gln Ser Val Gln Ser
Asp Gln Gly Tyr Ile Ser Arg Ser785 790 795 800Ser Pro Gln Pro Pro
Glu Gly Leu Thr Glu Met Glu Glu Glu Glu Glu 805 810 815Glu Glu Gln
Asp Pro Gly Lys Pro Ala Leu Pro Leu Ser Pro Glu Asp 820 825 830Leu
Glu Ser Leu Arg Ser Leu Gln Arg Gln Leu Leu Phe Arg Gln Leu 835 840
845Gln Lys Asn Ser Gly Trp Asp Thr Met Gly Ser Glu Ser Glu Gly Pro
850 855 860Ser Ala8652212PRTHomo sapiens 2Arg Ser Cys Asp Lys Thr
His Thr Cys Pro Pro Cys Pro Ala Pro Glu1 5 10 15Leu Leu Gly Gly Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 20 25 30Thr Leu Met Ile
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 35 40 45Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 50 55 60Val Glu
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn65 70 75
80Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
85 90 95Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
Pro 100 105 110Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
Pro Arg Glu 115 120 125Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp
Glu Leu Thr Lys Asn 130 135 140Gln Val Ser Leu Thr Cys Leu Val Lys
Gly Phe Tyr Pro Ser Asp Ile145 150 155 160Ala Val Glu Trp Glu Ser
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 165 170 175Thr Pro Pro Val
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 180 185 190Leu Thr
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 195 200
205Ser Val Met His 210331PRTHomo sapien 3Lys Gln Ile Glu Asp Lys
Ile Glu Glu Ile Leu Ser Lys Ile Tyr His1 5 10 15Ile Glu Asn Glu Ile
Ala Arg Ile Lys Lys Leu Ile Gly Glu Arg 20 25 30431PRTHomo sapiens
4Lys Gln Leu Glu Asp Lys Leu Glu Glu Leu Leu Ser Lys Leu Tyr His1 5
10 15Leu Glu Asn Glu Leu Ala Arg Leu Lys Lys Leu Leu Gly Glu Arg 20
25 30
* * * * *