U.S. patent application number 17/144685 was filed with the patent office on 2021-05-06 for treatment for rheumatoid arthritis.
This patent application is currently assigned to MORPHOSYS AG. The applicant listed for this patent is MORPHOSYS AG. Invention is credited to Bodo BROCKS, Daniela Della DUCATA, Stefan HARTLE, Stephane LECLAIR, Kai ROSPORT, Amgad SHEBL, Stefan STEIDL.
Application Number | 20210130451 17/144685 |
Document ID | / |
Family ID | 1000005331555 |
Filed Date | 2021-05-06 |
![](/patent/app/20210130451/US20210130451A1-20210506\US20210130451A1-2021050)
United States Patent
Application |
20210130451 |
Kind Code |
A1 |
HARTLE; Stefan ; et
al. |
May 6, 2021 |
TREATMENT FOR RHEUMATOID ARTHRITIS
Abstract
The present invention provides anti-GM-CSF antibodies for use in
the treatment of rheumatoid arthritis. Anti-GM-CSF antibodies, in
particular MOR103, are administered to patients suffering from
rheumatoid arthritis at dosages that are beneficial in a clinical
setting.
Inventors: |
HARTLE; Stefan; (Planegg,
DE) ; LECLAIR; Stephane; (Planegg, DE) ;
SHEBL; Amgad; (Planegg, DE) ; STEIDL; Stefan;
(Planegg, DE) ; BROCKS; Bodo; (Planegg, DE)
; DUCATA; Daniela Della; (Planegg, DE) ; ROSPORT;
Kai; (Planegg, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MORPHOSYS AG |
Planegg |
|
DE |
|
|
Assignee: |
MORPHOSYS AG
Planegg
DE
|
Family ID: |
1000005331555 |
Appl. No.: |
17/144685 |
Filed: |
January 8, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
15880726 |
Jan 26, 2018 |
10913792 |
|
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17144685 |
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14429996 |
Mar 20, 2015 |
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PCT/EP2013/069501 |
Sep 19, 2013 |
|
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15880726 |
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61703871 |
Sep 21, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 2039/505 20130101;
A61K 2039/54 20130101; A61K 39/3955 20130101; C07K 2317/76
20130101; C07K 16/243 20130101; A61K 31/519 20130101; A61K 2039/545
20130101; C07K 2317/21 20130101 |
International
Class: |
C07K 16/24 20060101
C07K016/24; A61K 31/519 20060101 A61K031/519; A61K 39/395 20060101
A61K039/395 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 20, 2012 |
EP |
12185235.4 |
Claims
1.-20. (canceled)
21. A pharmaceutical composition comprising: (a) an
anti-Granulocyte-Macrophage Colony Stimulating Factor (anti-GM-CSF)
antibody; (b) histidine; (c) sorbitol; and (d) Tween-80, wherein
said anti-GM-CSF antibody comprises the HCDR1 region of sequence
GFTFSSYWMN (SEQ ID NO.: 2), the HCDR2 region of sequence
GIENKYAGGATYYAASVKG (SEQ ID NO.: 3), the HCDR3 region of sequence
GFGTDF (SEQ ID NO.: 4), the LCDR1 region of sequence SGDSIGKKYAY
(SEQ ID NO.: 5), the LCDR2 region of sequence KKRPS (SEQ ID NO.:
6), and the LCDR3 region of sequence SAWGDKGM (SEQ ID NO.: 7).
22. The pharmaceutical composition according to claim 21, wherein
said anti-GM-CSF antibody comprises the variable heavy chain
peptide of sequence: TABLE-US-00004 (SEQ ID NO: 8)
QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMNWVRQAPGKGLEWVSG
IENKYAGGATYYAASVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
GFGTDFWGQGTLVTVSS;
and the variable light chain peptide of sequence: TABLE-US-00005
(SEQ ID NO: 9) DIELTQPPSVSVAPGQTARISCSGDSIGKKYAYWYQQKPGQAPVLVIYKK
RPSGIPERFSG SNSGNTATLTISGTQAEDEADYYCSAWGDKGMVFGGGT KLTVLGQ.
23. The pharmaceutical composition according to claim 21,
comprising: (b) 30 mM histidine; (c) 200 mM sorbitol; and (d) 0.02%
Tween-80.
24. The pharmaceutical composition according to claim 23,
comprising 40 mg to 400 mg of the anti-GM-CSF antibody.
25. The pharmaceutical composition according to claim 23,
comprising 75 mg, 100 mg, 150 mg, 180 mg, 200 mg, 300 mg, or 400 mg
of the anti-GM-CSF antibody.
26. The pharmaceutical composition according to claim 23,
comprising 75 mg of the anti-GM-CSF antibody.
27. The pharmaceutical composition according to claim 23,
comprising 100 mg of the anti-GM-CSF antibody.
28. The pharmaceutical composition according to claim 23,
comprising 150 mg of the anti-GM-CSF antibody.
29. The pharmaceutical composition according to claim 23,
comprising 180 mg of the anti-GM-CSF antibody.
30. The pharmaceutical composition according to claim 23, wherein
the composition has a pH of 6.0.
31. The pharmaceutical composition according to claim 22,
comprising: (b) 30 mM histidine; (c) 200 mM sorbitol; and (d) 0.02%
Tween-80.
32. The pharmaceutical composition according to claim 31,
comprising 40 mg to 400 mg of the anti-GM-CSF antibody.
33. The pharmaceutical composition according to claim 31,
comprising 75 mg, 100 mg, 150 mg, 180 mg, 200 mg, 300 mg, or 400 mg
of the anti-GM-CSF antibody.
34. The pharmaceutical composition according to claim 31,
comprising 75 mg of the anti-GM-CSF antibody.
35. The pharmaceutical composition according to claim 31,
comprising 100 mg of the anti-GM-CSF antibody.
36. The pharmaceutical composition according to claim 31,
comprising 150 mg of the anti-GM-CSF antibody.
37. The pharmaceutical composition according to claim 31,
comprising 180 mg of the anti-GM-CSF antibody.
38. The pharmaceutical composition according to claim 31, wherein
the composition has a pH of 6.0.
Description
FIELD OF THE INVENTION
[0001] The present invention provides anti-GM-CSF antibodies for
use in the treatment of rheumatoid arthritis, and methods for the
treatment of rheumatoid arthritis using such antibodies.
Anti-GM-CSF antibodies, in particular MOR103, are administered to
patients suffering from rheumatoid arthritis at dosages that are
beneficial in a clinical setting.
BACKGROUND OF THE INVENTION
[0002] Rheumatoid arthritis (RA) is a chronic systemic inflammatory
disease that affects 0.5% to 1% of the adult population worldwide.
RA primarily affects the joints and is characterized by chronic
inflammation of the synovial tissue, which eventually leads to the
destruction of cartilage, bone and ligaments and can cause joint
deformity. RA has a peak incidence between 40 and 60 years of age
and affects primarily women. The cause of RA is not known; however,
certain histocompatibility antigens are associated with poorer
outcomes. Nonsteroidal anti-inflammatory drugs (NSAIDs) provide
only symptomatic relief. Disease-modifying antirheumatic drugs
(DMARDs), the cornerstone of RA treatment throughout all stages of
the disease, maintain or improve physical function and retard
radiographic joint damage. Pro-inflammatory cytokines, such as
tumor necrosis factor-alpha (TNF.alpha.), interleukin-1,
interleukin-6 and granulocyte macrophage colony stimulating factor
(GM-CSF), which lead to the activation and proliferation of immune
cells, are found to be increased in the inflamed joint.
[0003] More recently, biological compounds, such as antibodies,
that target tumor necrosis factor alpha (TNF.alpha.), B-cells, or
T-cells have been used to treat RA, but still many patients fail to
respond to these therapies. Colony-stimulating factors (CSFs) have
been suggested for a potential point of intervention for
inflammatory disorders, such as RA (reviewed e.g. in Nat Rev
Immunol (2008) 8, 533-44) or Nat Rev Rheumatol (2009) 5, 554-9).
One of such CSF is granulocyte-macrophage colony-stimulation factor
(GM-CSF).
[0004] MOR103 is a fully human anti-GM-CSF antibody (Mol Immunol
(2008) 46, 135-44; WO 2006/122797). MOR 103 is also in a clinical
Phase Ib trial for multiple sclerosis. The present invention
describes the development of a clinically efficacious treatment
regimen comprising MOR103 for RA.
SUMMARY OF THE INVENTION
[0005] In one aspect, the present invention provides an anti-GM-CSF
antibody for use in the treatment of a patient suffering from
rheumatoid arthritis, wherein said antibody is administered to said
patient in a manner to achieve a therapeutically effective antibody
level in the blood of said patient equal or higher compared to the
intravenous administration of said antibody at a dose of at least
1.0 mg/kg when administered weekly over at least four weeks.
[0006] In another aspect, the present invention also provides a
method to treat a patient suffering from rheumatoid arthritis, said
method comprising administering to said patient an anti-GM-CSF
antibody in a manner to achieve a therapeutically effective
antibody level in the blood of said patient equal or higher
compared to the intravenous administration of said antibody at a
dose of at least 1.0 mg/kg when administered weekly over at least
four weeks.
[0007] In an embodiment, the anti-GM-CSF antibody is administered
intravenously, optionally at a dosage of at least 1.0 mg/kg, or at
a dose of about 1.0 mg/kg or about 1.5 mg/kg. In an embodiment, the
anti-GM-CSD antibody is administered weekly, over at least four
weeks.
[0008] In an embodiment, the anti-GM-CSF antibody is administered
subcutaneously, optionally at a dose of at least 2.0 mg/kg, or at a
dose of about 2.0 mg/kg, about 3.0 mg/kg or about 4.0 mg/kg. In an
embodiment, the anti-GM-CSF antibody is administered biweekly,
monthly or bimonthly. In another embodiment, the antibody is
administered at a fixed dose of about 75 mg, of about 100 mg, of
about 150 mg, of about 200 mg, of about 300 mg or of about 400 mg.
Administration of fixed doses may be every week, every second week,
every third week, every fourth week or every sixth week.
[0009] In an embodiment, the dosage of anti-GM-CSF antibody
administered to said patient and frequency of said administration
is sufficient to provide and maintain a serum concentration of said
antibody at at least 2 .mu.g/ml in said patient over the duration
of said treatment.
[0010] In another aspect, the present invention provides an
anti-GM-CSF antibody, wherein said anti-GM-CSF antibody is an
antibody comprising an HCDR1 region of sequence GFTFSSYWMN (SEQ ID
NO.: 2), an HCDR2 region of sequence GIENKYAGGATYYAASVKG (SEQ ID
NO.: 3), an HCDR3 region of sequence GFGTDF (SEQ ID NO.: 4), an
LCDR1 region of sequence SGDSIGKKYAY (SEQ ID NO.: 5), an LCDR2
region of sequence KKRPS (SEQ ID NO.: 6), and an LCDR3 region of
sequence SAWGDKGM (SEQ ID NO.: 7) for use in the treatment of a
patient suffering from rheumatoid arthritis, wherein said antibody
is administered to said patient in a manner to achieve a
therapeutically effective antibody level in the blood of said
patient equal or higher compared to the intravenous administration
of said antibody at a dose of at least 1.0 mg/kg when administered
weekly over at least four weeks.
[0011] In another aspect, the present invention provides an
anti-GM-CSF antibody, wherein said anti-GM-CSF antibody is an
antibody comprising an HCDR1 region of sequence GFTFSSYWMN (SEQ ID
NO.: 2), an HCDR2 region of sequence GIENKYAGGATYYAASVKG (SEQ ID
NO.: 3), an HCDR3 region of sequence GFGTDF (SEQ ID NO.: 4), an
LCDR1 region of sequence SGDSIGKKYAY (SEQ ID NO.: 5), an LCDR2
region of sequence KKRPS (SEQ ID NO.: 6), and an LCDR3 region of
sequence SAWGDKGM (SEQ ID NO.: 7) for use in the treatment of a
patient suffering from rheumatoid arthritis, wherein said antibody
is administered intravenously at a dose of about 1.0 mg/kg or at a
dose of about 1.5 mg/kg and wherein said antibody in administered
weekly over at least four weeks.
[0012] In another aspect, the present invention provides an
anti-GM-CSF antibody for use in the treatment of a patient
suffering from rheumatoid arthritis, wherein said antibody is
administered to said patient in a manner to achieve a
therapeutically effective antibody level in the blood of said
patient equal or higher compared to the intravenous administration
of said antibody at a dose of at least 1.0 mg/kg or at least 1.5
mg/kg when administered weekly over at least four weeks, and
wherein said anti-GM-CSF antibody is administered in combination
with a DMARD, such as methotrexate.
[0013] In an embodiment, the administration of said antibody to
achieve such a therapeutically effective amount comprises the
administration of said antibody intravenously at a dose at least
0.6, at least 0.7, at least 0.8, at least 0.9 or at least 1.0
mg/kg. In other embodiments, the antibody of the present invention
is administered intravenously at a dose of about 1.0 mg/kg or a
dose of about 1.5 mg/kg. Administration may be monthly, biweekly
(every two weeks) or weekly.
[0014] In another aspect, the present invention provides an
anti-GM-CSF antibody for use in the treatment of a patient
suffering from rheumatoid arthritis, wherein said antibody is
administered to said patient subcutaneously in a manner to achieve
a therapeutically effective antibody level in the blood of said
patient equal or higher compared to the intravenous administration
of said antibody at a dose of at least 1.0 mg/kg or at least 1.5
mg/kg when administered weekly over at least four weeks, and
wherein said anti-GM-CSF antibody is administered in combination
with a DMARD, such as methotrexate.
[0015] In an embodiment, the administration of said antibody to
achieve such a therapeutically effective amount comprises the
administration of said antibody subcutaneously at a dose of at
least 1.0, at least 1.5, at least 2.0, at least 2.5, at least 3.0,
at least 3.5 or at least 4.0 mg/kg. In other embodiments, the
antibody of the present invention is administered subcutaneously at
a dose of about 2.0 mg/kg, a dose of about 3.0 mg/kg or a dose of
about 4.0 mg/kg. Administration may be monthly, biweekly (every two
weeks) or weekly.
[0016] In an embodiment, the administration of said antibody to
achieve such a therapeutically effective amount comprises the
administration of said antibody subcutaneously at a fixed dose of
about 40 mg, at a fixed dose of 75 mg, at a fixed dose of 100 mg,
at a fixed dose of 140 mg, at a fixed dose of 150 mg, at a fixed
dose of 180 mg, at a fixed dose of 200 mg, at a fixed dose of 280
mg, at a fixed dose of 300 mg or at a fixed dose of 400 mg.
Administration of fixed doses may be every week, every second week,
every third week, every fourth week or every sixth week.
[0017] In another aspect, the present invention provides a method
of treating a patient suffering from rheumatoid arthritis, said
method comprising administering to said patient an anti-GM-CSF
antibody subcutaneously at [0018] (i) a dose of at least 1.0 mg/kg,
or [0019] (ii) a fixed dose of between 40 mg and 400 mg. The
anti-GM-CSF antibody may be administered to said patient in a
manner to achieve to a serum concentration of said antibody at at
least 2 .mu.g/ml in said patient over the duration of said
treatment. The antibody may be administered to said patient in a
manner to achieve a therapeutically effective antibody level in the
blood of said patient equal or higher compared to the intravenous
administration of said antibody at a dose of at least 1.0 mg/kg
when administered weekly over at least four weeks.
[0020] In another aspect, the present invention provides an
anti-GM-CSF antibody for inhibiting progression of structural joint
damage in a rheumatoid arthritis patient comprising administering
to said patient said antibody in a manner to achieve a
therapeutically effective antibody level in the blood of said
patient equal or higher compared to the intravenous administration
of said antibody at a dose of at least 1.0 mg/kg when administered
weekly over at least four weeks.
BRIEF DESCRIPTION OF THE DRAWINGS
[0021] FIG. 1 shows the amino acid sequence and the DNA sequence of
MOR04357.
[0022] FIG. 2 shows the mean changes of the DAS28 score after four
weeks (left panel) and after eight weeks (right panel) of treatment
compared. DAS28 score changes are compared to baseline levels, i.e.
disease status prior to treatment.
[0023] FIG. 3 shows the average ACR20 score of all treatment arms
after four weeks. An increase of the ACR20 scores corresponds to an
improvement of the severity of disease.
[0024] FIG. 4 shows the average ACR20 score of all treatment arms
after eight weeks. An increase of the ACR20 scores corresponds to
an improvement of the severity of disease.
DESCRIPTION
[0025] The terms "GM-CSF" and "GMCSF" refer to the protein known as
GM-CSF or Granulocyte-macrophage colony-stimulating factor, having
the following synonyms: Colony-stimulating factor 2, CSF2, GMCSF,
GM-CSF, Granulocyte-macrophage colony-stimulating factor,
MGC131935, MGC138897, Molgramostin, Sargramostim. Human GM-CSF has
the amino acid sequence of (UniProt P04141):
TABLE-US-00001 (SEQ ID NO.: 1)
MWLQSLLLLGTVACSISAPARSPSPSTQPWEHVNAIQEARRLLNLSRDTA
AEMNETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASH
YKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFDCWEPVQE
[0026] "MOR103" is an anti-GM-CSF antibody whose amino acid
sequence and DNA sequence is provided in FIG. 1. "MOR103" and
"MOR04357" and "MOR4357" are used as synonyms to describe the
antibody shown in FIG. 1. MOR04357 comprises an HCDR1 region of
sequence GFTFSSYWMN (SEQ ID NO.: 2), an HCDR2 region of sequence
GIENKYAGGATYYAASVKG (SEQ ID NO.: 3), an HCDR3 region of sequence
GFGTDF (SEQ ID NO.: 4), an LCDR1 region of sequence SGDSIGKKYAY
(SEQ ID NO.: 5), an LCDR2 region of sequence KKRPS (SEQ ID NO.: 6),
and an LCDR3 region of sequence SAWGDKGM (SEQ ID NO.: 7). MOR04357
comprises a variable heavy chain of the sequence
QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMNWVRQAPGKGLEWVSGIENKYAGGA
TYYAASVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGFGTDFWGQGTLVTVSS (SEQ ID
NO.: 8) and a variable light chain of the sequence
DIELTQPPSVSVAPGQTARISCSGDSIGKKYAYWYQQKPGQAPVLVIYKKRPSGIPERFSGS
NSGNTATLTISGTQAEDEADYYCSAWGDKGMVFGGGTKLTVLGQ (SEQ ID NO.: 9).
[0027] In certain embodiments, the antibody used in the present
invention is an antibody specific for GM-CSF. In other embodiments,
the antibody used in the present invention is an antibody specific
for a polypeptide encoding an amino acid sequence comprising SEQ ID
NO.: 1.
[0028] As used herein, "specifically for" or "specifically binding
to" refers to an antibody selectively or preferentially binding to
GM-CSF. Preferably the binding affinity for antigen is of Kd value
of 10.sup.-9 mol/l or lower (e.g. 10.sup.-10 mol/l), preferably
with a Kd value of 10.sup.-10 mol/l or lower (e.g. 10.sup.-12
mol/l). The binding affinity is determined with a standard binding
assay, such as surface plasmon resonance technique
(BIACORE.RTM.).
[0029] In certain embodiments, the antibody used in the present
invention is MOR103. In other embodiments, the antibody used in the
present invention is an antibody comprising an HCDR1 region of
sequence GFTFSSYWMN (SEQ ID NO.: 2), an HCDR2 region of sequence
GIENKYAGGATYYAASVKG (SEQ ID NO.: 3), an HCDR3 region of sequence
GFGTDF (SEQ ID NO.: 4), an LCDR1 region of sequence SGDSIGKKYAY
(SEQ ID NO.: 5), an LCDR2 region of sequence KKRPS (SEQ ID NO.: 6),
and an LCDR3 region of sequence SAWGDKGM (SEQ ID NO.: 7). In other
embodiments, the antibody used in the present invention is an
antibody comprising a variable heavy chain of the sequence
QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMNWVVRQAPGKGLEWVSGIENKYAGGA
TYYAASVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGFGTDFWGQGTLVTVSS (SEQ ID
NO.: 8) and a variable light chain of the sequence
DIELTQPPSVSVAPGQTARISCSGDSIGKKYAYWYQQKPGQAPVLVIYKKRPSGIPERFSGS
NSGNTATLTISGTQAEDEADYYCSAWGDKGMVFGGGTKLTVLGQ (SEQ ID NO.: 9). In
other embodiments, the antibody used in the present invention is an
antibody which cross-competes with an antibody comprising an HCDR1
region of sequence GFTFSSYWMN (SEQ ID NO.: 2), an HCDR2 region of
sequence GIENKYAGGATYYAASVKG (SEQ ID NO.: 3), an HCDR3 region of
sequence GFGTDF (SEQ ID NO.: 4), an LCDR1 region of sequence
SGDSIGKKYAY (SEQ ID NO.: 5), an LCDR2 region of sequence KKRPS (SEQ
ID NO.: 6), and an LCDR3 region of sequence SAWGDKGM (SEQ ID NO.:
7). In other embodiments, the antibody used in the present
invention is an antibody which binds to the same epitope like an
antibody specific for GM-CSF comprising an HCDR1 region of sequence
GFTFSSYWMN (SEQ ID NO.: 2), an HCDR2 region of sequence
GIENKYAGGATYYAASVKG (SEQ ID NO.: 3), an HCDR3 region of sequence
GFGTDF (SEQ ID NO.: 4), an LCDR1 region of sequence SGDSIGKKYAY
(SEQ ID NO.: 5), an LCDR2 region of sequence KKRPS (SEQ ID NO.: 6),
and an LCDR3 region of sequence SAWGDKGM (SEQ ID NO.: 7).
[0030] The term "antibody" is used in the broadest sense and
specifically covers monoclonal antibodies, polyclonal antibodies,
multispecific antibodies (e.g. bispecific antibodies) formed from
at least two intact antibodies, and antibody fragments so long as
they exhibit the desired biological activity.
[0031] "Antibody fragments" herein comprise a portion of an intact
antibody which retains the ability to bind antigen. Examples of
antibody fragments include Fab, Fab', F(ab')2, and Fv fragments;
diabodies; linear antibodies; single-chain antibody molecules; and
multispecific antibodies formed from antibody fragments.
[0032] The term "monoclonal antibody" as used herein refers to an
antibody obtained from a population of substantially homogeneous
antibodies, i.e., the individual antibodies comprising the
population are identical and/or bind the same epitope, except for
possible variants that may arise during production of the
monoclonal antibody, such variants generally being present in minor
amounts. In contrast to polyclonal antibody preparations that
typically include different antibodies directed against different
determinants (epitopes), each monoclonal antibody is directed
against a single determinant on the antigen. In addition to their
specificity, the monoclonal antibodies are advantageous in that
they are uncontaminated by other immunoglobulins.
[0033] The monoclonal antibodies herein specifically include
"chimeric" antibodies (immunoglobulins) in which a portion of the
heavy and/or light chain is identical with or homologous to
corresponding sequences in antibodies derived from a particular
species or belonging to a particular antibody class or subclass,
while the remainder of the chain(s) is identical with or homologous
to corresponding sequences in antibodies derived from another
species or belonging to another antibody class or subclass, as well
as fragments of such antibodies, so long as they exhibit the
desired biological activity.
[0034] "Humanized" forms of non-human (e.g., murine) antibodies are
chimeric antibodies that contain minimal sequence derived from
non-human immunoglobulin. For the most part, humanized antibodies
are human immunoglobulins (recipient antibody) in which residues
from a hypervariable region of the recipient are replaced by
residues from a hypervariable region of a non-human species (donor
antibody) such as mouse, rat, rabbit or nonhuman primate having the
desired specificity, affinity, and capacity. In some instances,
framework region (FR) residues of the human immunoglobulin are
replaced by corresponding non-human residues. Furthermore,
humanized antibodies may comprise residues that are not found in
the recipient antibody or in the donor antibody. These
modifications are made to further refine antibody performance. In
general, the humanized antibody will comprise substantially all of
at least one, and typically two, variable domains, in which all or
substantially all of the hypervariable regions correspond to those
of a non-human immunoglobulin and all or substantially all of the
FRs are those of a human immunoglobulin sequence, except for FR
substitution(s) as noted above. The humanized antibody optionally
also will comprise at least a portion of an immunoglobulin constant
region, typically that of a human immunoglobulin.
[0035] A "human antibody" herein is one comprising an amino acid
sequence structure that corresponds with the amino acid sequence
structure of an antibody obtainable from a human B-cell, and
includes antigen-binding fragments of human antibodies. Such
antibodies can be identified or made by a variety of techniques,
including, but not limited to: production by transgenic animals
(e.g., mice) that are capable, upon immunization, of producing
human antibodies in the absence of endogenous immunoglobulin;
selection from phage display libraries expressing human antibodies
or human antibody; generation via in vitro activated B; and
isolation from human antibody producing hybridomas.
[0036] In certain embodiments, the antibody used in the present
invention is a monoclonal antibody.
[0037] In other embodiments, the antibody used in the present
invention is a chimeric, a humanized or a human antibody. In
preferred embodiments, the antibody used in the present invention
is a human antibody.
[0038] In certain embodiments, the antibody used in the present
invention is administered in combination with an additional drug
that treats RA.
[0039] The additional drug may be one or more medicaments, and
include, for example, immunosuppressive agents, non-steroidal
anti-inflammatory drugs (NSAIDs), disease modifying anti-rheumatic
drugs (DMARDs) such as methotrexate (MTX), anti-B-cell surface
marker antibodies, such as anti-CD20 antibodies (e.g. rituximab),
TNF-alpha-inhibitors, corticosteroids, and co-stimulatory
modifiers, or any combination thereof. Optionally, the second or
additional drug is selected from the group consisting of
non-biological DMARDs, NSAIDS, and corticosteroids.
[0040] These additional drugs are generally used in the same
dosages and with administration routes as used hereinbefore and
hereinafter. If such additional drugs are used at all, preferably,
they are used in lower amounts than if the first medicament were
not present, especially in subsequent dosings beyond the initial
dosing with the first medicament, so as to eliminate or reduce side
effects caused thereby. The combined administration of an
additional drug includes co-administration (concurrent
administration), using separate formulations or a single
pharmaceutical formulation, and consecutive administration in
either order, wherein preferably there is a time period while both
(or all) active agents (medicaments) simultaneously exert their
biological activities.
[0041] The term "DMARD" refers to "Disease-Modifying Anti-Rheumatic
Drugs" and includes among others hydroxycloroquine, sulfasalazine,
methotrexate, leflunomide, azathioprine, D-penicillamine, gold
salts (oral), gold salts (intramuscular), minocycline, cyclosporine
including cyclosporine A and topical cyclosporine, and
TNF-inhibitors, including salts, variants, and derivatives thereof.
Exemplary DMARDs herein are non-biological, i.e. classic, DMARDs,
including, azathioprine, chloroquine, hydroxychloroquine,
leflunomide, methotrexate and sulfasalazine.
[0042] Methotrexate is an especially preferred DMARD of the present
invention. Therefore, in certain embodiments, the antibody used in
the present invention is administered in combination with a DMARD.
In other embodiments, the antibody used in the present invention is
administered in combination with methotrexate.
[0043] A "TNF-inhibitor" as used herein refers to an agent that
inhibits, to some extent, a biological function of TNF-alpha,
generally through binding to TNF-alpha and/or its receptor and
neutralizing its activity. Examples of TNF inhibitors include
etanercept (ENBREL.RTM.), infliximab (REMICADE.RTM.), adalimumab
(HUMIRA.RTM.), certolizumab pegol (CIMZIA.RTM.), and golimumab
(SIMPONI.RTM.).
[0044] "Treatment" of a patient or a subject refers to both
therapeutic treatment and prophylactic or preventative measures.
The terms "effective amount" or "therapeutically effective" refer
to an amount of the antibody that is effective for treating
rheumatoid arthritis. Such effective amount can result in any one
or more of reducing the signs or symptoms of RA (e.g. achieving
ACR20), reducing disease activity (e.g. Disease Activity Score,
DAS20), slowing the progression of structural joint damage or
improving physical function. In one embodiment, such clinical
response is comparable to that achieved with intravenously
administered anti-GM-CSF antibody.
[0045] The antibody of the present invention may be administered in
different suitable forms. Potential forms of administration include
systemic administration (subcutaneous, intravenous, intramuscular),
oral administration, inhalation, transdermal administration,
topical application (such as topical cream or ointment, etc.) or by
other methods known in the art. The doses (in mg/kg) specified in
the present invention refer to milligrams of antibody per kilogram
of body weight of the patient. In vitro cell based assays showed
that an anti-GM-CSF antibody (MOR103) is capable of inhibiting
several GM-CSF mediated responses. Evaluated responses include TF-1
cell proliferation, STAT5 phosphorylation, polymorphonuclear
neutrophils (PMN) migration, PMN up-regulation of CD11b, monocyte
up-regulation of MHC II, and eosinophil survival. Complete
inhibitory effects were generally reached at concentrations of
about 0.2 .mu.g/ml anti-GM-CSF antibody. GM-CSF concentrations up
to 1 ng/ml were applied in such studies. As a reference, GM-CSF
levels in the synovial fluid of RA patients were reported to be
<500 pg/ml. It is reasonable to consider that similar GM-CSF
concentrations as used in these in vitro studies are present in
affected tissues of RA patients
[0046] To effectively treat RA it may be important for an
anti-GM-CSF antibody to penetrate the synovium. There is evidence
to suggest that monoclonal antibodies can distribute into the
synovium when dosed subcutaneously or intravenously. Based on a
predicted penetration rate of 30%, continuous GM-CSF production and
considering patient heterogeneity, the minimal or sub-optimal
clinical effect level in RA patients is anticipated to be at a
serum concentration of approximately 2 .mu.g/ml antibody (thus,
approximately 10-fold higher than the inhibitory concentration
derived from in vitro studies).
[0047] A specific anti-GM-CSF antibody (MOR103) has been
administered to patients with active rheumatoid arthritis who
received 4 intravenous weekly doses of 0.3, 1, and 1.5 mg/kg. The
anti-GM-CSF antibody showed significant clinical efficacy on DAS28,
EULAR, ACR20, ACR50, ACR70 and tender joint counts following once a
week dosing with 1 and 1.5 mg/kg as compared to placebo.
[0048] In certain embodiments, the antibody of the present
invention is administered intravenously. In other embodiments, the
antibody of the present invention is administered
subcutaneously.
[0049] From other therapeutic antibodies it is known that a
concentration that leads to a certain level of the antibody in the
blood when administered intravenously corresponds to about 50-76%
of the blood concentration achieved when the same antibody
concentration is administered subcutaneously (Meibohm, B.:
Pharmacokinetics and Pharmacodynamics of Biotech Drugs, Wiley-VCH,
2006). For MOR103 this ratio was determined to be 52%, i.e. a given
concentration administered subcutaneously leads to a blood
concentration which is equivalent to about 52% of the blood
concentration when the same given concentration is administered
intravenously. Therefore, the concentration of a subcutaneous
formulation needs to be about twice as high to achieve the same
drug blood level as compared to an intravenous formulation.
[0050] In certain embodiments the blood level to be achieved in a
patient is equal or higher compared to the blood concentration
achieved with intravenous administration of the antibody of the
present invention at a dose of at least 1.0 mg/kg when administered
weekly over at least four weeks.
[0051] In alternative embodiments said blood concentration to be
achieved is equal or higher compared to the blood concentration
achieved with intravenous administration of the antibody of the
present invention at a doses of at least 0.4, 0.5, 0.6, 0.7, 0.8 or
0.9 mg/kg when administered weekly over at least four weeks. In
alternative embodiments the blood level to be achieved in a patient
is equal or higher compared to the blood concentration achieved
with intravenous administration of the antibody of the present
invention at a dose of at least 1.0 mg/kg when administered weekly
over at least two weeks or at least three weeks. In alternative
embodiments the blood level to be achieved in a patient is equal or
higher compared to the blood concentration achieved with
intravenous administration of the antibody of the present invention
at a dose of at least 1.0 mg/kg when administered biweekly over at
least two weeks or at least four weeks.
[0052] In certain embodiments, the antibody of the present
invention is administered intravenously. In other embodiments, the
antibody of the present invention is administered intravenously at
a dose at least 0.6, at least 0.7, at least 0.8, at least 0.9 or at
least 1.0 mg/kg. In other embodiments, the antibody of the present
invention is administered intravenously at a dose of about 1.0
mg/kg or a dose of about 1.5 mg/kg.
[0053] In certain embodiments, the antibody of the present
invention is administered subcutaneously. Various dosing regimen
have been simulated using the subcutaneous delivery of MOR103 in
order to produce plasma concentrations that are similar those
obtained after 1 mg/kg iv, a dose that was efficacious in RA. The
majority of simulations produce trough concentration values greater
than 2 ug/mL, a value that is believed to be the minimum blood
concentration that is required to produce efficacy in the context
of an anti-GM-CSF antibody. These studies indicate that
subcutaneous doses of 1, 2, 3 and 4 mg/kg can produce plasma
concentration similar to 1 mg/kg, IV depending on the dosing
frequency.
[0054] In other embodiments, the antibody of the present invention
is administered subcutaneously at a dose at least 1.0, at least
1.5, at least 2.0, at least 2.5, at least 3.0, at least 3.5 or at
least 4.0 mg/kg. In other embodiments, the antibody of the present
invention is administered subcutaneously at a dose of about 2.0
mg/kg, a dose of about 3.0 mg/kg or a dose of about 4.0 mg/kg. In
certain embodiments, the antibody of the present invention is
subcutaneously administered biweekly, monthly or bimonthly.
[0055] In other embodiments, the antibody of the present invention
is administered subcutaneously at a fixed dose. In such "fixed
dose" treatment the antibody is administered at a certain, fixed,
concentration, i.e. without taking into account a patient's body
weight. In certain embodiments, the antibody of the present
invention is administered at a fixed dose of between 40 mg and 400
mg, optionally at a fixed dose of 75 mg, at a fixed dose of 100 mg,
at a fixed dose of 140 mg, at a fixed dose of 150 mg, at a fixed
dose of 180 mg, at a fixed dose of 200 mg, at a fixed dose of 280
mg, at a fixed dose of 300 mg or at a fixed dose of 400 mg.
Administration of fixed doses may be every week, every second week,
every third week, every fourth week or every sixth week. Typically,
the antibody will be administered weekly at a fixed dose.
[0056] In an embodiment, the antibody will be administered weekly,
at a fixed subcutaneous dose of 40, 56, 70, 75 100, 140, 150, 180,
200, 210, or 280 mg.
[0057] In an embodiment, the antibody will be administered
biweekly, at a fixed subcutaneous dose of 70, 75, 100, 140, 150,
180, 200, 210, 280 or 300 mg.
[0058] In an embodiment, the antibody will be administered monthly,
at a fixed subcutaneous dose of 100, 140, 150, 180, 200, 210, 280,
300, 320, 350. 360 or 400 mg.
[0059] In an embodiment, the antibody is administered in a dose
sufficient to maintain trough concentration of antibody of at least
2 ug/mL. The trough concentration of antibody may be maintained at
2.0 ug/mL, 2.5 ug/mL, 3.0 ug/mL, 3.5 ug/mL, 4.0 ug/mL, 4.5 ug/mL or
5.0 ug/mL, during the course of therapy.
[0060] In alternative embodiments, the antibody will be
administered weekly, at a fixed subcutaneous dose of 28 or 35
mg.
[0061] In certain embodiments, the present invention provides an
anti-GM-CSF antibody for use in the treatment of a patient
suffering from rheumatoid arthritis, wherein said antibody is
administered to said patient in a manner to achieve a
therapeutically effective antibody level in the blood of said
patient equal or higher compared to the intravenous administration
of said antibody at a dose of at least 1.0 mg/kg when administered
weekly over at least four weeks.
[0062] In certain embodiments, the present invention provides a
method to treat a patient suffering from rheumatoid arthritis, said
method comprising administering to said patient an anti-GM-CSF
antibody in a manner to achieve a therapeutically effective
antibody level in the blood of said patient equal or higher
compared to the intravenous administration of said antibody at a
dose of at least 1.0 mg/kg when administered weekly over at least
four weeks.
[0063] In certain embodiments, the present invention provides an
anti-GM-CSF antibody for inhibiting progression of structural joint
damage in a rheumatoid arthritis patient comprising administering
to said patient said antibody in a manner to achieve a
therapeutically effective antibody level in the blood of said
patient equal or higher compared to the intravenous administration
of said antibody at a dose of at least 1.0 mg/kg when administered
weekly over at least four weeks.
[0064] The terms "drug" and "medicament" refer to an active drug to
treat rheumatoid arthritis or joint damage or symptoms or side
effects associated with RA. The term "pharmaceutical formulation"
refers to a preparation which is in such form as to permit the
biological activity of the active ingredient or ingredients, i.e.
the antibody of the present invention, to be effective, and which
contains no additional components which are unacceptably toxic to a
subject to which the formulation would be administered. Such
formulations are sterile.
[0065] The antibody herein is preferably recombinantly produced in
a host cell transformed with nucleic acid sequences encoding its
heavy and light chains (e.g. where the host cell has been
transformed by one or more vectors with the nucleic acid therein).
The preferred host cell is a mammalian cell, most preferably a
PER.C6 cell.
[0066] Therapeutic formulations of the antibody of the present
invention are prepared for storage by mixing the antibody having
the desired degree of purity with optional pharmaceutically
acceptable carriers, excipients or stabilizers in the form of
lyophilized formulations or aqueous solutions. Acceptable carriers,
excipients, or stabilizers are nontoxic to recipients at the
dosages and concentrations employed, and include buffers such as
phosphate, citrate, histidine and other organic acids; antioxidants
including ascorbic acid and methionine; preservatives (such as
octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride;
benzalkonium chloride, benzethonium chloride; phenol, butyl or
benzyl alcohol; alkyl parabens such as methyl or propyl paraben;
catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low
molecular weight (less than about 10 residues) polypeptides;
proteins, such as serum albumin, gelatin, or immunoglobulins;
hydrophilic polymers such as polyvinylpyrrolidone; amino acids such
as glycine, glutamine, asparagine, histidine, arginine, or lysine;
monosaccharides, disaccharides, and other carbohydrates including
glucose, mannose, or dextrins; chelating agents such as EDTA;
sugars such as sucrose, mannitol, trehalose or sorbitol;
salt-forming counter-ions such as sodium; metal complexes {e.g.
Zn-protein complexes); and/or non-ionic surfactants such as
TWEEN.TM. (such as Tween-80), PLURONICS.TM. or polyethylene glycol
(PEG).
[0067] In certain embodiments, the present invention provides a
pharmaceutical composition comprising an antibody of the present
invention and a pharmaceutically acceptable carrier and/or
excipient for use in any of the methods provided in the present
invention. In certain embodiments, the formulation for the antibody
of the present invention consists of 30 mM histidine, pH 6.0, 200
mM sorbitol and 0.02% Tween-80. In other embodiments, the
formulation for the antibody of the present invention consists of
PBS, pH 6.2 (0.2 g/l KCl, 0.96 g/l KH.sub.2PO.sub.4, 0.66 g/l
Na.sub.2HPO.sub.4.times.7H.sub.2O, 8 g/l NaCl).
EXAMPLES
Example 1: Design and Concept of a Clinical Phase Lb/Phase Ila
Trial
[0068] A multi-center, randomized, double-blinded,
placebo-controlled study to evaluate the safety, preliminary
clinical activity and immunogenicity of multiple doses of MOR103
administered intravenously to patients with active rheumatoid
arthritis was conducted.
[0069] Primary outcome measures were the adverse event rate and the
safety profile. Secondary outcome measures included DAS28 scores,
ACR scores and EULAR28 response criteria.
[0070] The clinical trial comprised three treatment arms. In each
treatment arm patient received either placebo or MOR103. The MOR103
doses were 0.3 mg/kg body weight for treatment arm 1, 1.0 mg/kg
body weight for treatment arm 2 and 1.5 mg/kg body weight for
treatment arm 3. MOR103 and placebo were administered
intravenously, weekly with 4 doses in total.
[0071] Summary of the treatment arms:
TABLE-US-00002 Arms Assigned Interventions Experimental: Group 1:
Drug: MOR103 MOR103, experimental MOR103 0.3 mg/kg or Biological:
MOR103 0.3 mg/kg placebo iv .times. 4 doses or placebo
Experimental: Group 2: Drug: MOR103 MOR103, experimental MOR103 1.0
mg/kg or Biological: MOR103 1.0 mg/kg placebo iv .times. 4 doses or
placebo Experimental: Group 3: Drug: MOR103 MOR103, experimental
MOR103 1.5 mg/kg or Biological: MOR103 1.5 mg/kg placebo iv .times.
4 doses or placebo
[0072] Eligible for participation in the study were patients of 18
years and older and of either sex (male and female). Healthy
volunteers were not accepted.
[0073] Inclusion criteria were as follows: [0074] Rheumatoid
arthritis (RA) per revised 1987 ACR criteria [0075] Active RA:
.gtoreq.3 swollen and 3 tender joints with at least 1 swollen joint
in the hand, excluding the PIP joints [0076] CRP>5.0 mg/L (RF
and anti-CCP seronegative); CRP>2 mg/l (RF and/or anti-CCP
seropositive) [0077] DAS28.ltoreq.5.1 [0078] Stable regimen of
concomitant RA therapy (NSAIDs, steroids, non-biological DMARDs).
[0079] Negative PPD tuberculin skin test
[0080] Exclusion criteria were as follows: [0081] Previous therapy
with B or T cell depleting agents other than Rituximab (e.g.
Campath). Prior treatment with Rituximab, TNF-inhibitors, other
biologics (e.g. anti-IL-1 therapy) and systemic immunosuppressive
agents is allowed with a washout period. [0082] Any history of
ongoing, significant or recurring infections [0083] Any active
inflammatory diseases other than RA [0084] Treatment with a
systemic investigational drug within 6 months prior to screening
[0085] Women of childbearing potential, unless receiving stable
doses of methotrexate or leflunomide [0086] Significant cardiac or
pulmonary disease (including methotrexate-associated lung toxicity)
[0087] Hepatic or renal insufficiency
Example 2: Patient Recruitment and Patient Population
[0088] Clinical sites for patient recruitment, screening and
treatment were located in Bulgaria, Germany, the Netherlands,
Poland and the Ukraine.
[0089] 96 patients were included in the trial. 27 patients received
placebo, 24 patients received MOR103 at a dose of 0.3 mg/kg, 22
patients received MOR103 at a dose of 1.0 mg/kg and 23 patients
received MOR103 at a dose of 1.5 mg/kg. The average age and the
average Body Mass Index (BMI) was about the same for all treatment
groups. Key characteristics are summarized in the following
Table:
TABLE-US-00003 MOR 103 Active Treatment Groups 0.3 1.0 1.5 Total
Placebo mg/kg mg/kg mg/kg active Characteristic N = 27 N = 24 N =
22 N = 23 N = 69 Age 53.8 57.4 49 53 53.3 BMI 26.3 26.3 26.1 25.7
26.0 Gender-Female 19 (70%) 21 (88%) 17 (77%) 18 (78%) 56 (81%)
White 27 24 22 23 69
[0090] 90% of all patients of the study were previously treated
with DMARDs. The most commonly used DMARD was methotrexate (75% of
all patients). The rate of previous treatment with DMARDs was
comparable in all treatment arms.
[0091] Prior to administration of MOR103 or the placebo the disease
activity of all patients was measured according to accepted
guidelines by calculating the DAS28 score, a 28-joint Disease
Activity Score (see e.g. Ann Rheum Dis (2009) 68, 954-60). DAS28
score is a validated and commonly used tool to quantify the disease
status of RA patients. The average DAS28 score was comparable for
all treatment arms.
Example 3: Safety Profile
[0092] Based on the available observed safety data, MOR103 showed a
favorable safety profile among all doses tested. The key
observations are as follows: [0093] No deaths were observed during
the conduct of the trial [0094] No infusion related reactions were
observed [0095] Two serious adverse events (SAEs) were observed:
[0096] One patient in the placebo group developed paronychia [0097]
One patient in the 0.3 mg/kg treatment arm developed pleurisy
[0098] More treatment-emergent adverse effects (TEAEs) were
observed in the placebo group (25.9%) than in the active groups
(14.5%) [0099] Most TEAEs were mild [0100] No severe TEAEs were
observed in the active groups
[0101] In summary, it can be concluded that treatment with MOR103
at all doses tested is safe. Two serious adverse events were
observed, both none in the treatment arms that showed clinical
efficacy (see below). Sub-cutaneous administration of MOR103 at a
dose that leads to an antibody drug level in the blood of patients
equivalent to the intravenous application of the present study is
expected to show a similar safety profile.
Example 4: Efficacy--DAS28
[0102] 4 weeks and 8 weeks after the first administration of MOR103
(or placebo) the DAS28 scores of all patients was determined. A
decrease in DAS28 scores correlates to diminished disease severity.
Results are shown in FIG. 2 as the mean changes compared to
baseline, i.e. disease status prior to treatment.
[0103] The placebo group only shows marginal changes. Patients
treated with MOR103 at 0.3 mg/kg showed a slight decrease in DAS28
scores, indicating slightly less severity of the disease. In
contrast, patients treated with MOR103 at 1.0 mg/kg or with 1.5
mg/kg showed a significant decrease in DAS28 scores, indicating the
high efficacy of MOR103 at these doses.
Example 5: Efficacy--ACR20
[0104] As another measure of efficacy the ACR20 criteria were used.
ACR criteria measure improvement in tender or swollen joint counts
and improvement in certain other parameters. The procedure to
measure ACR scores is highly standardized. The present clinical
trial applied the respective applicable guidelines. Results are
depicted in FIGS. 3 and 4. A higher score corresponds to an
improvement in the severity of the disease.
[0105] In line with the results of the DAS28 scores (see Example
4), also the ACR scores show a strong clinical improvement of
patients' condition upon treatment with either 1.0 mg/kg MOR103 or
1.5 mg/kg MOR103. The improvement after 4 weeks is highly
significant for the 1.0 mg/kg group (p<0.0001). Taken together,
the ACR20 scores confirm the surprising finding that the efficacy
of MOR103 can already be shown with a comparably low number or
patients in each treatment arm and a comparably short treatment
period.
Example 6: Clinical Trial with Additional Doses of MOR103
[0106] The clinical trial set out herein above is repeated with
additional doses of MOR103. MOR103 is administered to patients
intravenously at a dose of 0.5 mg/kg (treatment arm 1) and 0.75
mg/kg (treatment arm 2). All other parameters are identical to
Example 1.
[0107] Both treatment arms show a favorable safety profile and
demonstrate clinical efficacy as measured by DAS28 scores and ACR20
scores.
Example 7: Clinical Trial with a Sub-Cutaneous Formulation of
MOR103
[0108] The clinical trial set out herein above is repeated with a
sub-cutaneous formulation of MOR103. In order to achieve similar
levels of MOR103 in the blood of patients as observed for
intravenous treatment, the sub-cutaneous dose of MOR103 is
increased.
[0109] In different treatment arms MOR103 is administered to
patients at 1.5 mg/kg, 2.0 mg/kg, 3.0 mg/kg and 4.0 mg/kg. The drug
is administered sub-cutaneously, either biweekly, monthly or
bimonthly. All other parameters are identical to Example 1.
[0110] All treatment arms show a favorable safety profile and
demonstrate clinical efficacy as measured by DAS28 scores and ACR20
scores.
Example 8: Clinical Trial with a Sub-Cutaneous Formulation of
MOR103 at a Fixed Dose
[0111] Example 7 is repeated with a fixed dose of MOR103. In
different treatment arms MOR103 is administered to patients at
fixed dose of 75 mg, of 100 mg, of 150 mg, of 200 mg, of 300 mg and
of 400 mg. The drug is administered sub-cutaneously every week,
every second week, every fourth week or every sixth week. All other
parameters are identical to the Examples described herein
above.
[0112] All treatment arms show a favorable safety profile and
demonstrate clinical efficacy as measured by DAS28 scores and ACR20
scores.
Sequence CWU 1
1
91144PRTHomo Sapiens 1Met Trp Leu Gln Ser Leu Leu Leu Leu Gly Thr
Val Ala Cys Ser Ile1 5 10 15Ser Ala Pro Ala Arg Ser Pro Ser Pro Ser
Thr Gln Pro Trp Glu His 20 25 30Val Asn Ala Ile Gln Glu Ala Arg Arg
Leu Leu Asn Leu Ser Arg Asp 35 40 45Thr Ala Ala Glu Met Asn Glu Thr
Val Glu Val Ile Ser Glu Met Phe 50 55 60Asp Leu Gln Glu Pro Thr Cys
Leu Gln Thr Arg Leu Glu Leu Tyr Lys65 70 75 80Gln Gly Leu Arg Gly
Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met 85 90 95Met Ala Ser His
Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser 100 105 110Cys Ala
Thr Gln Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys 115 120
125Asp Phe Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu
130 135 140210PRTArtificial Sequencepolypeptide 2Gly Phe Thr Phe
Ser Ser Tyr Trp Met Asn1 5 10319PRTArtificial Sequencepolypeptide
3Gly Ile Glu Asn Lys Tyr Ala Gly Gly Ala Thr Tyr Tyr Ala Ala Ser1 5
10 15Val Lys Gly46PRTArtificial Sequencepolypeptide 4Gly Phe Gly
Thr Asp Phe1 5511PRTArtificial Sequencepolypeptide 5Ser Gly Asp Ser
Ile Gly Lys Lys Tyr Ala Tyr1 5 1065PRTArtificial
Sequencepolypeptide 6Lys Lys Arg Pro Ser1 578PRTArtificial
Sequencepolypeptide 7Ser Ala Trp Gly Asp Lys Gly Met1
58117PRTArtificial Sequencepolypeptide 8Gln Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Trp Met Asn Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Glu
Asn Lys Tyr Ala Gly Gly Ala Thr Tyr Tyr Ala Ala 50 55 60Ser Val Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr65 70 75 80Leu
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90
95Tyr Cys Ala Arg Gly Phe Gly Thr Asp Phe Trp Gly Gln Gly Thr Leu
100 105 110Val Thr Val Ser Ser 1159106PRTArtificial
Sequencepolypeptide 9Asp Ile Glu Leu Thr Gln Pro Pro Ser Val Ser
Val Ala Pro Gly Gln1 5 10 15Thr Ala Arg Ile Ser Cys Ser Gly Asp Ser
Ile Gly Lys Lys Tyr Ala 20 25 30Tyr Trp Tyr Gln Gln Lys Pro Gly Gln
Ala Pro Val Leu Val Ile Tyr 35 40 45Lys Lys Arg Pro Ser Gly Ile Pro
Glu Arg Phe Ser Gly Ser Asn Ser 50 55 60Gly Asn Thr Ala Thr Leu Thr
Ile Ser Gly Thr Gln Ala Glu Asp Glu65 70 75 80Ala Asp Tyr Tyr Cys
Ser Ala Trp Gly Asp Lys Gly Met Val Phe Gly 85 90 95Gly Gly Thr Lys
Leu Thr Val Leu Gly Gln 100 105
* * * * *