U.S. patent application number 15/235263 was filed with the patent office on 2017-01-05 for arthritis treatment.
The applicant listed for this patent is XBiotech, Inc.. Invention is credited to John Simard.
Application Number | 20170002071 15/235263 |
Document ID | / |
Family ID | 45328882 |
Filed Date | 2017-01-05 |
United States Patent
Application |
20170002071 |
Kind Code |
A1 |
Simard; John |
January 5, 2017 |
Arthritis Treatment
Abstract
Administration of a monoclonal Ab (mAb) that specifically
targets IL-1.alpha. is useful to treating articular and
extra-articular symptoms of arthritis.
Inventors: |
Simard; John; (Austin,
TX) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
XBiotech, Inc. |
Vancouver |
|
CA |
|
|
Family ID: |
45328882 |
Appl. No.: |
15/235263 |
Filed: |
August 12, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13162705 |
Jun 17, 2011 |
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15235263 |
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61356176 |
Jun 18, 2010 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 19/02 20180101;
A61P 19/00 20180101; A61K 2039/505 20130101; A61P 27/02 20180101;
A61P 29/00 20180101; C07K 16/245 20130101 |
International
Class: |
C07K 16/24 20060101
C07K016/24 |
Claims
1. A method for treating pain associated with arthritis in a human
subject, the method comprising: providing a pharmaceutical
composition comprising a pharmaceutically acceptable carrier and an
anti-IL-1.alpha. antibody, and administering the pharmaceutical
composition to the subject in an amount effective to reduce the
pain associated with arthritis in the subject, wherein the pain is
reduced.
2. The method of claim 1, wherein the pain occurs in a joint of the
subject.
3. The method of claim 1, wherein the pain is in the uvea of the
subject.
4. The method of claim 1, wherein the anti-IL-1.alpha. antibody is
a monoclonal antibody.
5. The method of claim 1, wherein the anti-IL-1.alpha. antibody is
a monoclonal antibody that comprises the all of the CDRs of
MABp1.
6. The method of claim 1, wherein the antibody is MABp1.
7. The method of claim 2, wherein the pharmaceutical composition is
administered by subcutaneous injection.
8. The method of claim 3, wherein the pharmaceutical composition is
administered to the eye topically.
9. The method of claim 1, wherein the amount of the
anti-IL-1.alpha. antibody effective to reduce pain associated with
arthritis in the subject is sufficient to raise the subject's
peripheral blood concentration of anti-IL-1.alpha. antibody to at
least 4 micrograms/ml.
10. The method of claim 1, wherein the amount of the
anti-IL-1.alpha. antibody effective to reduce pain associated with
arthritis in the subject is sufficient to raise monocyte
vacuolization in the subject.
11. The method of claim 2, wherein the mobility of the joint is
improved after administration of the pharmaceutical
composition.
12. The method of claim 2, wherein the pharmaceutical composition
is administered to the subject after an injection of corticosteroid
failed to relieve the pain.
13. The method of claim 1, wherein the subject lacked detectable
endogenous anti-IL-la antibodies before administration of the
pharmaceutical composition.
14. The method of claim 1, wherein the pharmaceutical composition
is repeatedly administered to the subject until the pain is
completely resolved.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of U.S.
patent application Ser. No. 13/162,705 filed on Jun. 17, 2011,
which claims priority from U.S. provisional patent application Ser.
No. 61/356,176 filed on Jun. 18, 2010. All patent applications are
incorporated herein by reference in their entirety.
STATEMENT AS TO FEDERALLY SPONSORED RESEARCH
[0002] Not applicable.
FIELD OF THE INVENTION
[0003] The invention relates generally to the fields of immunology,
inflammation, arthritis, and medicine. More particularly, the
invention relates to the use of antibodies (Abs) which specifically
bind interleukin-1.alpha. (IL-1.alpha.) to treat one or more
symptoms of arthritis.
BACKGROUND
[0004] Arthritis, the most common cause of disability in the United
States, is a collection of different conditions such as
osteoarthritis, rheumatoid arthritis, gout, psoriatic arthritis,
septic arthritis, and reactive arthritis. All types of arthritis
are characterized by joint inflammation which causes pain,
swelling, redness, stiffness, and warmth at that affected site.
Because afflicted subjects are less mobile due to pain and
stiffness, arthritis can indirectly lead to obesity, high
cholesterol, and/or heart disease. Arthritis can also cause
extra-articular disease such as iritis, uveitis, oral ulcers,
inflammation of the gastrointestinal tract, inflammation of the
genitourinary tract, and skin lesions.
[0005] For most types of arthritis, no cure exists and treatment is
largely symptomatic, e.g., administration of analgesics and
anti-inflammatory drugs. Non-steroidal anti-inflammatory drugs
(NSAIDs) can be used to reduce inflammation and pain. While
generally effective, NSAIDs may cause side effects such as
abdominal pain, bleeding, ulcers, and liver and kidney damage.
Corticosteroids are effective at reducing inflammation and joint
damage, but can cause a number of side effect are also associated
including bruising, weight gain, cataracts, bone thinning,
diabetes, and hypertension. Other drugs commonly used to treat
arthritis are methotrexate, cyclosporine, cyclophosphamide,
leflunomide, hydroxychloroquine, sulfasalazine, and minocycline.
These too can cause side effects such as liver damage and
immunosuppression. Tumor necrosis factor (TNF) inhibitors like
etanercept (Enbrel), infliximab (Remicade), and adalimumab (Humira)
are also useful for treating arthritis. Side effects of TNF
inhibitors include injection site reactions, heart failure,
lymphoma, and increased risk of infection.
SUMMARY
[0006] The invention is based on the discovery that administration
of an antibody (Ab) that specifically targets IL-1.alpha. in a
human subject suffering from arthritis reduces the number of
CD14+IL-1.alpha.+ peripheral blood monocytes in the subject and
markedly ameliorates inflammation in both articular and
extraarticular sites--all without any observed side effects other
than pain at the administration site.
[0007] Accordingly, the invention features a method of treating an
inflammatory pathology associated with arthritis in a human subject
by administering to the subject a pharmaceutical composition
including a pharmaceutically acceptable carrier and an amount of an
anti-IL-1.alpha. antibody effective to reduce at least one symptom
of the inflammatory pathology in the subject. The symptom can be
joint inflammation such as of the wrist or shoulder, or
inflammation of the eye such as uveitis. The anti-IL-1.alpha.
antibody can be a monoclonal antibody such as an IgG1. The
anti-IL-1.alpha. antibody can be the monoclonal antibody designated
as MABp1 or a monoclonal antibody that includes one or more
complementarity determining regions (CDRs) of MABp1.
[0008] The pharmaceutical composition can be administered to the
subject by injection, subcutaneously, intravenously,
intramuscularly, intraocularly, or directly into an inflamed joint.
The antibody might also be administered to the eye topically. In
the method, the amount of the anti-IL-1.alpha. antibody effective
to reduce at least one symptom of the inflammatory pathology in the
subject can be sufficient to raise the subject's peripheral blood
concentration of anti-IL-1.alpha. antibody to at least 4 ug/ml;
and/or sufficient to decrease the number of the subject's
CD14+IL-1.alpha.+ peripheral blood monocytes by at least 5%.
[0009] The method might also include a step of measuring the number
of CD14+IL-1.alpha.+ monocytes in the subject's peripheral blood
after administration of the pharmaceutical composition, e.g.,
wherein the step of measuring the number of CD14+IL-1.alpha.+
monocytes in the subject's peripheral blood is performed at least
two different time points after administration of the
pharmaceutical composition.
[0010] In another aspect, the invention features a method inducing
monocyte vacuolization in a subject by administering to the subject
a pharmaceutical composition including a pharmaceutically
acceptable carrier and an amount of an anti-IL-1.alpha. antibody
effective to induce vacuole formation in monocytes.
[0011] Unless otherwise defined, all technical terms used herein
have the same meaning as commonly understood by one of ordinary
skill in the art to which this invention belongs. Commonly
understood definitions of biological terms can be found in Rieger
et al., Glossary of Genetics: Classical and Molecular, 5th edition,
Springer-Verlag: New York, 1991; and Lewin, Genes V, Oxford
University Press: New York, 1994. Commonly understood definitions
of medical terms can be found in Stedman's Medical Dictionary,
27.sup.th Edition, Lippincott, Williams & Wilkins, 2000.
[0012] As used herein, an "antibody" or "Ab" is an immunoglobulin
(Ig), a solution of identical or heterogeneous Igs, or a mixture of
Igs. An "antibody" can also refer to fragments and engineered
versions of Igs such as Fab, Fab', and F(ab')2 fragments; and
scFv's, heteroconjugate Abs, and similar artificial molecules that
employ Ig-derived CDRs to impart antigen specificity. A "monoclonal
antibody" or "mAb" is an Ab expressed by one clonal B cell line or
a population of Ab molecules that contains only one species of an
antigen binding site capable of immunoreacting with a particular
epitope of a particular antigen. A "polyclonal antibody" or
"polyclonal Ab" is a mixture of heterogeneous Abs. Typically, a
polyclonal Ab will include myriad different Ab molecules which bind
a particular antigen with at least some of the different Abs
immunoreacting with a different epitope of the antigen. As used
herein, a polyclonal Ab can be a mixture of two or more mAbs.
[0013] An "antigen-binding portion" of an Ab is contained within
the variable region of the Fab portion of an Ab and is the portion
of the Ab that confers antigen specificity to the Ab (i.e.,
typically the three-dimensional pocket formed by the CDRs of the
heavy and light chains of the Ab). A "Fab portion" or "Fab region"
is the proteolytic fragment of a papain-digested Ig that contains
the antigen-binding portion of that Ig. A "non-Fab portion" is that
portion of an Ab not within the Fab portion, e.g., an "Fc portion"
or "Fc region." A "constant region" of an Ab is that portion of the
Ab outside of the variable region. Generally encompassed within the
constant region is the "effector portion" of an Ab, which is the
portion of an Ab that is responsible for binding other immune
system components that facilitate the immune response. Thus, for
example, the site on an Ab that binds complement components or Fc
receptors (not via its antigen-binding portion) is an effector
portion of that Ab.
[0014] When referring to a protein molecule such as an Ab,
"purified" means separated from components that naturally accompany
such molecules. Typically, an Ab or protein is purified when it is
at least about 10% (e.g., 9%, 10%, 20%, 30% 40%, 50%, 60%, 70%,
80%, 90%, 95%, 98%, 99%, 99.9%, and 100%), by weight, free from the
non-Ab proteins or other naturally-occurring organic molecules with
which it is naturally associated. Purity can be measured by any
appropriate method, e.g., column chromatography, polyacrylamide gel
electrophoresis, or HPLC analysis. A chemically-synthesized protein
or other recombinant protein produced in a cell type other than the
cell type in which it naturally occurs is "purified."
[0015] By "bind", "binds", or "reacts with" is meant that one
molecule recognizes and adheres to a particular second molecule in
a sample, but does not substantially recognize or adhere to other
molecules in the sample. Generally, an Ab that "specifically binds"
another molecule has a Kd greater than about 10.sup.5, 10.sup.6,
10.sup.7, 10.sup.8, 10.sup.9, 10.sup.10, 10.sup.11, or 10.sup.12
liters/mole for that other molecule.
[0016] A "therapeutically effective amount" is an amount which is
capable of producing a medically desirable effect in a treated
animal or human (e.g., amelioration or prevention of a disease or
symptom of a disease).
[0017] Although methods and materials similar or equivalent to
those described herein can be used in the practice or testing of
the present invention, suitable methods and materials are described
below. All publications mentioned herein are incorporated by
reference in their entirety. In the case of conflict, the present
specification, including definitions will control. In addition, the
particular embodiments discussed below are illustrative only and
not intended to be limiting.
BRIEF DESCRIPTION OF THE DRAWINGS
[0018] FIG. 1 is a graph and table showing the pharmacokinetics of
MABp1 after administration to a human subject with reactive
arthritis.
[0019] FIG. 2 is a series of graphs and histograms showing flow
cytometric blood analyses after administration of MABp1 to a human
subject with reactive arthritis.
[0020] FIG. 3 is a series of graphs showing flow cytometric blood
analyses after administration of MABp1 to a human subject with
reactive arthritis.
DETAILED DESCRIPTION
[0021] The invention encompasses compositions and methods for
treating a symptom or pathologic process associated with arthritis
in a subject. The below described preferred embodiments illustrate
adaptation of these compositions and methods. Nonetheless, from the
description of these embodiments, other aspects of the invention
can be made and/or practiced based on the description provided
below.
General Methodology
[0022] Methods involving conventional immunological and molecular
biological techniques are described herein. Immunological methods
(for example, assays for detection and localization of antigen-Ab
complexes, immunoprecipitation, immunoblotting, and the like) are
generally known in the art and described in methodology treatises
such as Current Protocols in Immunology, Coligan et al., ed., John
Wiley & Sons, New York. Techniques of molecular biology are
described in detail in treatises such as Molecular Cloning: A
Laboratory Manual, 2nd ed., vol. 1-3, Sambrook et al., ed., Cold
Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 2001; and
Current Protocols in Molecular Biology, Ausubel et al., ed., Greene
Publishing and Wiley-Interscience, New York. Ab methods are
described in Handbook of Therapeutic Abs, Dubel, S., ed.,
Wiley-VCH, 2007. General methods of medical treatment are described
in McPhee and Papadakis, Current Medical Diagnosis and Treatment
2010, 49.sup.th Edition, McGraw-Hill Medical, 2010; and Fauci et
al., Harrison's Principles of Internal Medicine, 17.sup.th Edition,
McGraw-Hill Professional, 2008
Treatment of Arthritis Symptoms
[0023] The compositions and methods described herein are useful for
treating an inflammatory pathology associated with arthritis in a
mammalian subject by administering to the subject a pharmaceutical
composition including an amount of an anti-IL-1.alpha. antibody
effective to reduce at least one symptom of the inflammatory
pathology in the subject. The mammalian subject might be any that
suffers from arthritis including, human beings, dogs, cats, horses,
cattle, sheep, goats, and pigs. Human subjects might be male,
female, adults, children, seniors (65 and older), and those with
other diseases. The particular symptom or pathologic process
associated with arthritis can be inflammation, pain, stiffness, or
degeneration of a joint (e.g., in the wrist, digits [metacarpal or
metatarsal joints], elbows, shoulders, hips, knees, ankles, foot,
neck, or back) or extraarticular tissue (e.g., iritis, uveitis,
oral ulcers, inflammation of the gastrointestinal tract,
inflammation of the genitourinary tract, or skin lesions).
Antibodies and Other Agents that Target IL-1.alpha.
[0024] Any suitable type of Ab that specifically binds IL-1.alpha.
and reduces a symptom or pathologic process caused by arthritis in
a subject might be used in the invention. For example, the
anti-IL-1.alpha. Ab used might be mAb, a polyclonal Ab, a mixture
of mAbs, or an Ab fragment or engineered Ab-like molecule such as
an scFv. The Ka of the Ab is preferably at least 1.times.10.sup.9
M.sup.-1 or greater (e.g., greater than 9.times.10.sup.10 M.sup.-1,
8.times.10.sup.10 M.sup.-1, 7.times.10.sup.10 M.sup.-1,
6.times.10.sup.10 M.sup.-1, 5.times.10.sup.10 M.sup.-1,
4.times.10.sup.10 M.sup.-1, 3.times.10.sup.10 M.sup.-1,
2.times.10.sup.10 M.sup.-1, or 1.times.10.sup.10 M.sup.-1). In a
preferred embodiment, the invention utilizes a fully human mAb that
includes (i) an antigen-binding variable region that exhibits very
high binding affinity for human IL-la and (ii) a constant region
that is effective at both activating the complement system though
C1q binding and binding to several different Fc receptors. The
human Ab is preferably an IgG1, although it might be of a different
isotype such as IgM, IgA, or IgE, or subclass such as IgG2, IgG3,
or IgG4. One example of a particularly useful mAb is MABp1, an
IL-1.alpha.-specific IgG1 monoclonal antibody described in U.S.
patent application Ser. No. 12/455,458 filed on Jun. 1, 2009. Other
useful mAbs are those than include at least one but preferably all
the CDRs of MABp1.
[0025] Because B lymphocytes which express Ig specific for human
IL-1.alpha. occur naturally in human beings, a presently preferred
method for raising mAbs is to first isolate such a B lymphocyte
from a subject and then immortalize it so that it can be
continuously replicated in culture. Subjects lacking large numbers
of naturally occurring B lymphocytes which express Ig specific for
human IL-1.alpha. may be immunized with one or more human
IL-1.alpha. antigens to increase the number of such B lymphocytes.
Human mAbs are prepared by immortalizing a human Ab secreting cell
(e.g., a human plasma cell). See, e.g., U.S. Pat. No.
4,634,664.
[0026] In an exemplary method, one or more (e.g., 5, 10, 25, 50,
100, 1000, or more) human subjects are screened for the presence of
such human IL-1.alpha.-specific Ab in their blood. Those subjects
that express the desired Ab can then be used as B lymphocyte
donors. In one possible method, peripheral blood is obtained from a
human donor that possesses B lymphocytes that express human
IL-1.alpha.-specific Ab. Such B lymphocytes are then isolated from
the blood sample, e.g., by cells sorting (e.g., fluorescence
activated cell sorting, "FACS"; or magnetic bead cell sorting) to
select B lymphocytes expressing human IL-1.alpha.-specific Ig.
These cells can then be immortalized by viral transformation (e.g.,
using EBV) or by fusion to another immortalized cell such as a
human myeloma according to known techniques. The B lymphocytes
within this population that express Ig specific for human
IL-1.alpha. can then be isolated by limiting dilution methods
(e.g., cells in wells of a microtiter plate that are positive for
Ig specific for human IL-1.alpha. are selected and subcultured, and
the process repeated until a desired clonal line can be isolated).
See, e.g., Goding, Monoclonal Abs: Principles and Practice, pp.
59-103, Academic Press, 1986. Those clonal cell lines that express
Ig having at least nanomolar or picomolar binding affinities for
human IL-1.alpha. are preferred. MAbs secreted by these clonal cell
lines can be purified from the culture medium or a bodily fluid
(e.g., ascites) by conventional Ig purification procedures such as
salt cuts, size exclusion, ion exchange separation, and affinity
chromatography.
[0027] Although immortalized B lymphocytes might be used in in
vitro cultures to directly produce mAbs, in certain cases it might
be desirable to use heterologous expression systems to produce
mAbs. See, e.g., the methods described in U.S. patent application
Ser. No. 11/754,899. For example, the genes encoding an mAb
specific for human IL-1.alpha. might be cloned and introduced into
an expression vector (e.g., a plasmid-based expression vector) for
expression in a heterologous host cell (e.g., CHO cells, COS cells,
myeloma cells, and E. coli cells). Because Igs include heavy (H)
and light (L) chains in an H2L2 configuration, the genes encoding
each may be separately isolated and expressed in different
vectors.
[0028] Although generally less preferred due to the greater
likelihood that a subject will develop an anti-Ab response,
chimeric mAbs (e.g., "humanized" mAbs), which are antigen-binding
molecules having different portions derived from different animal
species (e.g., variable region of a mouse Ig fused to the constant
region of a human Ig), might be used in the invention. Such
chimeric Abs can be prepared by methods known in the art. See,
e.g., Morrison et al., Proc. Nat'l. Acad. Sci. USA, 81:6851, 1984;
Neuberger et al., Nature, 312:604, 1984; Takeda et al., Nature,
314:452, 1984. Similarly, Abs can be humanized by methods known in
the art. For example, monoclonal Abs with a desired binding
specificity can be humanized by various vendors or as described in
U.S. Pat. Nos. 5,693,762; 5,530,101; or 5,585,089.
[0029] The mAbs described herein might be affinity matured to
enhance or otherwise alter their binding specificity by known
methods such as VH and VL domain shuffling (Marks et al.
Bio/Technology 10:779-783, 1992), random mutagenesis of the
hypervariable regions (HVRs) and/or framework residues (Barbas et
al. Proc Nat. Acad. Sci. USA 91:3809-3813, 1994; Schier et al. Gene
169:147-155, 1995; Yelton et al. J. Immunol. 155:1994-2004, 1995;
Jackson et al., J. Immunol. 154(7):3310-9, 1995; and Hawkins et al,
J. Mol. Biol. 226:889-896, 1992. Amino acid sequence variants of an
Ab may be prepared by introducing appropriate changes into the
nucleotide sequence encoding the Ab. In addition, modifications to
nucleic acid sequences encoding mAbs might be altered (e.g.,
without changing the amino acid sequence of the mAb) for enhancing
production of the mAb in certain expression systems (e.g., intron
elimination and/or codon optimization for a given expression
system). The mAbs described herein can also be modified by
conjugation to another protein (e.g., another mAb) or non-protein
molecule. For example, a mAb might be conjugated to a water soluble
polymer such as polyethylene glycol or a carbon nanotube (See,
e.g., Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605,
2005). See, U.S. patent application Ser. No. 11/754,899.
[0030] Preferably, to ensure that high titers of human
IL-1.alpha.-specific mAb can be administered to a subject with
minimal adverse effects, the mAb compositions of the invention are
at least 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 95, 96, 97, 98, 99,
99.9 or more percent by weight pure (excluding any excipients). The
mAb compositions of the invention might include only a single type
of mAb (i.e., one produced from a single clonal B lymphocyte line)
or might include a mixture of two or more (e.g., 2, 3, 4, 5, 6, 7,
8, 9, 10 or more) different types of mAbs.
[0031] To modify or enhance their function, the human IL-1.alpha.
mAbs might be conjugated another molecule such as a cytotoxin. A
human IL-1.alpha. specific mAb might be conjugated with one or more
cytotoxins to more effectively kill cells expressing IL-1.alpha..
Cytotoxins for use in the invention can be any cytotoxic agent
(e.g., molecule that can kill a cell after contacting the cell)
that can be conjugated to a human IL-1.alpha. specific mAb.
Examples of cytotoxins include, without limitation, radionuclides
(e.g., .sup.35S, .sup.14C, .sup.32P, .sup.125I, .sup.131I,
.sup.90Y, .sup.89Zr, .sup.201Tl, .sup.186Re, .sup.188Re, .sup.57Cu,
.sup.213Bi, and .sup.211At), conjugated radionuclides, and
chemotherapeutic agents. Further examples of cytotoxins include,
but are not limited to, antimetabolites (e.g., 5-fluorouricil
(5-FU), methotrexate (MTX), fludarabine, etc.), anti-microtubule
agents (e.g., vincristine, vinblastine, colchicine, taxanes (such
as paclitaxel and docetaxel), etc.), alkylating agents (e.g.,
cyclophasphamide, melphalan, bischloroethylnitrosurea (BCNU),
etc.), platinum agents (e.g., cisplatin (also termed cDDP),
carboplatin, oxaliplatin, JM-216, CI-973, etc.), anthracyclines
(e.g., doxorubicin, daunorubicin, etc.), antibiotic agents (e.g.,
mitomycin-C), topoisomerase inhibitors (e.g., etoposide,
tenoposide, and camptothecins), or other cytotoxic agents such as
ricin, diptheria toxin (DT), Pseudomonas exotoxin (PE) A, PE40,
abrin, saporin, pokeweed viral protein, ethidium bromide,
glucocorticoid, anthrax toxin and others. See, e.g., U.S. Pat. No.
5,932,188.
[0032] While the IL-1.alpha. specific Abs described above are
preferred for use the invention, in some cases, other agents that
specifically target IL-1.alpha. might be used so long as their
administration leads to improvement of one or more symptoms of
arthritis. These other agents might include small organic
molecules, aptamers, peptides, and proteins that specifically bind
IL-1.alpha..
Pharmaceutical Compositions and Methods
[0033] The anti-IL-1.alpha. Ab compositions may be administered to
animals or humans in pharmaceutically acceptable carriers (e.g.,
sterile saline), that are selected on the basis of mode and route
of administration and standard pharmaceutical practice. A list of
pharmaceutically acceptable carriers, as well as pharmaceutical
formulations, can be found in Remington's Pharmaceutical Sciences,
a standard text in this field, and in USP/NF. Other substances may
be added to the compositions and other steps taken to stabilize
and/or preserve the compositions, and/or to facilitate their
administration to a subject.
[0034] For example, the Ab compositions might be lyophilized (see
Draber et al., J. Immunol. Methods. 181:37, 1995; and
PCT/US90/01383); dissolved in a solution including sodium and
chloride ions; dissolved in a solution including one or more
stabilizing agents such as albumin, glucose, maltose, sucrose,
sorbitol, polyethylene glycol, and glycine; filtered (e.g., using a
0.45 and/or 0.2 micron filter); contacted with beta-propiolactone;
and/or dissolved in a solution including a microbicide (e.g., a
detergent, an organic solvent, and a mixture of a detergent and
organic solvent.
[0035] The Ab compositions may be administered to animals or humans
by any suitable technique. Typically, such administration will be
parenteral (e.g., intravenous, subcutaneous, intramuscular, or
intraperitoneal introduction). The compositions may also be
administered directly to the target site (e.g., an inflamed joint,
or the uvea or conjuctiva) by, for example, injection or topical
application. Other methods of delivery, e.g., liposomal delivery or
diffusion from a device impregnated with the composition, are known
in the art. The composition may be administered in a single bolus,
multiple injections, or by continuous infusion (e.g., intravenously
or by peritoneal dialysis).
[0036] A therapeutically effective amount is an amount which is
capable of producing a medically desirable result in a treated
animal or human. An effective amount of anti-IL-la Ab compositions
is an amount which shows clinical efficacy in arthritis patients as
measured by the improvement in pain and function as well as the
prevention of structural damage. As is well known in the medical
arts, dosage for any one animal or human depends on many factors,
including the subject's size, body surface area, age, the
particular composition to be administered, sex, time and route of
administration, general health, and other drugs being administered
concurrently. A preferred dose is one that is sufficient to raise
the subject's peripheral blood concentration of anti-IL-1.alpha. Ab
to at least 4 (e.g., at least 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50,
100, 200, 300, 400, 500, 1000, 2500, or 5000) micrograms/ml. It is
expected that an appropriate dosage of Abs would be in the range of
about 0.2 to 20 (e.g., 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20,
30, 50, or 100) mg/kg body weight for subcutaneous administration
and about 0.001 to 50 (e.g., 0.001, 0.01, 1, 5, 10, 15, 25, or 50)
mg per eye for topical administration to the eye. The dose may be
given repeatedly, e.g., hourly, daily, weekly, or monthly.
EXAMPLES
Example 1
Xilonix.TM.
[0037] Xilonix.TM. is a sterile injectable liquid formulation of 15
mg/mL MABp1 in a stabilizing isotonic buffer (pH 6.4). Each 10-mL
Type I borosilicate glass serum vial contains 5 mL of the
formulation, and is sealed with a 20-mm Daikyo Flurotec butyl
rubber stopper and flip-off aluminum seal. The product is stored at
5.+-.3.degree. C., with excursions to room temperature permitted.
The exact composition of the drug product is shown below:
TABLE-US-00001 Composition of the Drug Product (Xilonix .TM.)
Ingredient Grade Manufacturer Concentration MABp1 antibody GMP
XBiotech 15 mg/mL sodium phosphate dibasic compendial JT Baker 12
mg/mL citric acid monohydrate compendial JT Baker 2 mg/mL
Trehalose.cndot.2H2O (high- compendial Ferro- 60 mg/mL purity low
endotoxin) Pfanstiehl polysorbate 80 compendial JT Baker 0.2 mg/mL
Phosphoric acid, to compendial JT Baker 0.04 mg/mL adjust pH water
for injection compendial Microbix q.s.
Example 2
Treatment of Reactive Arthritis with an IL-1.alpha.-Specific
Monoclonal Antibody
[0038] A 48 year-old male patient with reactive arthritis was
administered a total 220 milligrams of MABp1, an
IL-1.alpha.-specific monoclonal antibody described in U.S. patent
application Ser. No. 12/455,458 filed on Jun. 1, 2009. The patient
had a long history of reactive arthritis, starting at age 16, when
he was diagnosed with Reiter's syndrome during hospitalization for
severe inflammation in his left knee. This inflammation resolved,
yet the patient experienced periodic relapses in several joints
until his mid-twenties. No further episodes occurred until, at age
35, the patient had a severe unilateral episode of uveitis that
lasted for 8 weeks. The uveitis was poorly managed with ophthalmic
corticosteroids and oral NSAIDS, resulting in some scaring. The
patient subsequently experienced at least three additional episodes
of uveitis of varying intensities, one episode requiring subcorneal
injection of corticosteroids.
[0039] Just prior to his 48.sup.th birthday, the patient developed
severe pain in his left shoulder and wrist. Evident swelling and
redness with almost complete loss of mobility affected the wrist.
The patient was unable to abduct his left arm greater than about
20.degree. due to intense shoulder pain. On that day, the patient
was given a subacromial injection of corticosteroids into the left
shoulder. The patient reported that the condition continued to
worsen with pain from shoulder and wrist reportedly becoming
continuous, interrupting work and preventing sleep. In addition,
pain and irritation in the left eye ensued, indicating onset of an
episode of uveitis. This was reportedly the first time joint
inflammation and uveitis occurred together. The patient was taking
ophthalmic corticosteroids, oral and topical ophthalmic NSAIDS with
little apparent benefit.
[0040] On day 0 (forty-two days after the subacromial injection of
corticosteroids), the patient was administered four subcutaneous
injections of MABp1, delivering a total of 110 mg of MABp1 (in
equal doses). No side effects other than pain during injection was
reported. Blood was drawn by venous puncture immediately prior to
injection into two 5 ml sodium heparin tubes. Plasma analysis using
an enzyme-linked immunadsorbant assay (ELISA) for the detection of
existing endogenous anti-IL-1.alpha. antibodies revealed no
pre-existing antibodies.
[0041] On day 1, the patient reported that he woke up that morning
without the throbbing pain that had become the "first sensation
upon waking." Over the next several days there was an evident
improvement in mobility. There was no induration or redness at the
injection sites. A blood draw was taken and flow cytometric
analysis (FACS) was performed to evaluate leuckocyte subsets and
IL-1.alpha. expression on monocytes. Analysis was also performed on
plasma to determine levels of MABp1 and to begin collection of
pharmacokinetic (pK) data for MABp1. FACS analysis of PBMC revealed
that most CD14+ monocytes (72.6%) expressed IL-1.alpha.. A MABp1
plasma concentration of 3.2 .mu.g/ml was observed.
[0042] On day 6, another blood sample was taken and analyzed using
FACS and for MABp1. The frequency of CD14+ monocytes stained by
MABp1 had declined to 47.3%. Plasma levels of MABp1 had increased
to 7 .mu.g/ml. Although not confirmed, the increase in MABp1
concentration was considered to reflect a depot effect of the
subcutaneous administration of MABp1. Although there had been
improvement, the patient still exhibited considerable tenderness
and pain with movement and the uveitis had flared since the
previous weekend, where the patient had attended a party and
consumed alcohol. The patient was administered another 110 mg of
MABp1 subcutaneously.
[0043] On day 14, a blood sample was taken and analyzed using FACS
and pK analysis was performed on plasma. CD14+ monocyte frequency
stained by MABp1 further declined to 21.7%. However, plasma levels
of MABp1 had also declined to 5.8 .mu.g/ml. This was unanticipated,
since plasma levels of MABp1 had increased over the week after the
first injection.
[0044] Approximately one month after the first injection of MABp1
the patient was reevaluated. Marked improvement was noted in
mobility and there was no pain in the wrist. Pain in shoulder was
present only upon abduction to 90.degree.. FACS analysis revealed
no detectable CD14+ monocytes stained by MABp1. Plasma levels of
MABp1 had declined to 1.6 .mu.g/ml, suggesting a half-life for
MABp1 of about two weeks.
[0045] Over the course of the next several weeks the patient showed
gradual but continuous improvement in mobility. There was complete
resolution of the uveitis. The improvement was noted even though
the patient discontinued use of all medications after the first
injection of MABp1. Approximately three months after the first
injection of MABp1, the frequency of CD14+ monocytes stained by
MABp1 had returned to pre-treatment levels. MABp1 levels in plasma
declined to 0.07 .mu.g/ml. However, the patient continued to do
well with continuing improvement in mobility of the shoulder.
Example 3
Screening of Plasma Samples for Endogenous Autoantibody Against
hIL-1A and Pharmacokinetics of MABp1
[0046] A method was developed for the screening of plasma samples
for endogenous autoantibody against human IL-1.alpha.
(hIL-1.alpha.) using a direct ELISA. This method was also used to
determine pharmacokinetics (pK) of MABp1 after administration, with
the exception, that higher dilutions plasma samples were made.
[0047] The direct ELISA involves coating of recombinant human
IL-1.alpha. on a polystyrene microplate. The bound human IL-1A
captures endogenous anti-human IL-1.alpha. antibody from test
samples. An HRP-conjugated-Fc specific, mouse-anti-human IgG is
then used to detect the captured endogenous anti-human IL-1A
antibody, followed by treatment with TMB substrate. On reacting
with HRP enzyme, the TMB substrate produces a deep blue-colored
soluble product. The enzymatic reaction is stopped by the addition
of a stop solution that turns the blue-colored product to yellow.
The colorimetric measurements are carried out on a microplate
reader at 450 nm.
[0048] About 5 ml plasma sample per sample is provided. Plasma is
kept at 2-8.degree. C. prior to aliquoting and storage at
-80.degree. C. Plasma samples are diluted 1:500, 1:1000 and
1:2000-fold to use as samples. A positive control in buffer is used
containing 20 .mu.g/ml MABp1 antibody stock as 1:5,000 and
1:10,000-fold dilutions on microplate. Buffer is used as a negative
control as well as a pre-determined negative control plasma, which
is diluted as 1:1,000, 1:2,000 and 1:5,000. An additional positive
plasma control is used, which is plasma spiked with 20 .mu.g/ml
MABp1 antibody and diluted as 1:5,000 and 1:10,000 for samples on
the microplate.
[0049] If the positive control value falls within .+-.2 standard
deviation, the ELISA data is considered acceptable. However if the
QC positive control value falls beyond .+-.2 standard deviation,
the ELISA data is considered unacceptable and the experiment would
be repeated. Using a Kaleidagraph, the logarithmic mean absorbance
of standard solution is plotted as a function of logarithmic
concentration along with absorbance error bars. The standard curve
should exhibit a linear behavior. Results from a pharmacokinetics
analysis of samples taken from the patient as described in Example
2 are shown in FIG. 1.
Example 4
Flow Cytometric (FACS) Examination of Blood Lineage Subsets
[0050] FACS procedures are described for both whole blood staining,
and staining of peripheral blood mononuclear cells (PBMC) enriched
from whole blood. Both whole blood and PBMC staining was performed
on all samples. This FACS analysis allows relative percentage
determination of blood lineage subsets: B and T lymphocytes, NK
cells, monocytes, neutrophils, and IL-1.alpha.+cells. Results from
FACs analyses of samples taken from the patient as described in
Example 2 are shown in FIGS. 2 and 3. A photomicrograph of a blood
smear showed that MABp1 administration caused extensive
vacuolization in peripheral blood monocytes when analyzed 32 days
post administration.
Example 5
Treatment of Uveitis with an IL-1.alpha.-Specific Monoclonal
Antibody
[0051] About two months following resolution of the uveitis
described in Example 2, the patient experienced another episode of
uveitis (predominantly iritis). The patient was started on
corticoseroid and non-steroidal anti-inflammatory drops (NSAIDS).
Oral NSAIDS were also used. The uveitis was unresponsive to
treatment and progressed. However, there was no evidence of any
joint involvement, with shoulder continuing to show improvement in
mobility. The patient was administered MABp1 topically to the
affected eye. MABp1 (15 mg/ml solution) was administered at a rate
of one drop per minute, for ten minutes, for a total of ten drops
to the affected eye (approximately 3.75 mg in 0.25 ml). The patient
did not complain of any pain during the administration. However,
for several hours after, the patient reported discomfort and
burning. Oral NSAIDs were taken and the patient slept. The next
morning, the patient reported considerable improvement, reduced
pain and less inflammation than prior to administration.
Twenty-four hours after the first administration of the MABP1
drops, the patient administered 10 drops in the same fashion.
Again, discomfort and burning was noted. Oral NSAIDs were taken,
and again the patient took bed rest. The uveitis resolved itself
completely. No further medications were taken. No recurrence of
uveitis was observed over the next four months.
Other Embodiments
[0052] It is to be understood that while the invention has been
described in conjunction with the detailed description thereof, the
foregoing description is intended to illustrate and not limit the
scope of the invention, which is defined by the scope of the
appended claims. Other aspects, advantages, and modifications are
within the scope of the following claims.
* * * * *