U.S. patent application number 11/929604 was filed with the patent office on 2008-02-28 for treatment of lfa-1 associated disorders with increasing doses of lfa-1 antagonist.
This patent application is currently assigned to Genentech, Inc.. Invention is credited to Russell L. Dedrick, Marvin R. Garovoy, Susan M. Kramer, Karen Starko.
Application Number | 20080050364 11/929604 |
Document ID | / |
Family ID | 27383218 |
Filed Date | 2008-02-28 |
United States Patent
Application |
20080050364 |
Kind Code |
A1 |
Garovoy; Marvin R. ; et
al. |
February 28, 2008 |
Treatment of LFA-1 Associated Disorders with Increasing Doses of
LFA-1 Antagonist
Abstract
A method is provided for reducing the occurrence of fever,
headache, nausea and/or vomiting associated with administration of
a therapeutic compound to a mammal in need thereof, comprising
administering to the mammal a first conditioning dose of a
non-target cell depleting compound which binds to a cell surface
receptor on a target mammalian cell; and administering a second
therapeutic dose of the compound, wherein the second dose is higher
than the first dose.
Inventors: |
Garovoy; Marvin R.; (San
Anselmo, CA) ; Kramer; Susan M.; (San Francisco,
CA) ; Dedrick; Russell L.; (Kensington, CA) ;
Starko; Karen; (Hillsborough, CA) |
Correspondence
Address: |
GENENTECH, INC.
1 DNA WAY
SOUTH SAN FRANCISCO
CA
94080
US
|
Assignee: |
Genentech, Inc.
South San Francisco
CA
94080
Xoma Ltd.
Berkeley
CA
94718
|
Family ID: |
27383218 |
Appl. No.: |
11/929604 |
Filed: |
October 30, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11078332 |
Mar 10, 2005 |
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11929604 |
Oct 30, 2007 |
|
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|
10655905 |
Sep 5, 2003 |
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11078332 |
Mar 10, 2005 |
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09936603 |
Feb 11, 2002 |
6652855 |
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PCT/US00/07189 |
Mar 17, 2000 |
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10655905 |
Sep 5, 2003 |
|
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60125351 |
Mar 19, 1999 |
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60125228 |
Mar 19, 1999 |
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Current U.S.
Class: |
424/130.1 ;
424/133.1 |
Current CPC
Class: |
A61K 39/39591 20130101;
A61K 31/395 20130101; C07K 16/2845 20130101; A61K 2039/505
20130101; C07K 2317/24 20130101; A61K 2039/545 20130101; A61P 43/00
20180101 |
Class at
Publication: |
424/130.1 ;
424/133.1 |
International
Class: |
A61K 31/395 20060101
A61K031/395; A61P 43/00 20060101 A61P043/00 |
Claims
1. A method for treating dermatitis, comprising administering to a
mammal in need thereof a first conditioning dose of an antibody or
a receptor binding fragment thereof which binds CD11a; and
administering a second therapeutic dose of the antibody, wherein
the second dose is higher than the first dose.
2. The method of claim 1 wherein the antibody is a humanized
antibody.
3. The method of claim 1, further comprising administering a third
therapeutic dose, wherein the third dose is higher than or equal to
the second dose.
4. The method of claim 1, further comprising administering a fourth
therapeutic dose, wherein the fourth dose is higher than or equal
to the third dose.
5. The method of claim 1 or claim 2, wherein administration is
intravenous or subcutaneous.
6. The method of claim 1 or claim 2, wherein administration is not
more than once per week.
7. The method of claim 1 or claim 2, wherein the antibody or
receptor binding fragment thereof is non-lymphocyte depleting.
8. The method of claim 7, wherein the antibody or receptor binding
fragment thereof is non-T-cell depleting.
9. The method of claim 1 or claim 2, wherein the first conditioning
dose is 0.7 mg/kg and the second therapeutic dose is 1.0 mg/kg.
10. The method of claim 1 or claim 2, wherein the first
conditioning dose is 0.7 mg/kg and the second therapeutic dose is
2.0 mg/kg.
11. A method for treating graft versus host or host versus graft
transplant rejection, comprising administering to a mammal in need
thereof a first conditioning dose of an antibody or a receptor
binding fragment thereof which binds CD11a; and administering a
second therapeutic dose of the antibody, wherein the second dose is
higher than the first dose.
12. The method of claim 11 wherein the antibody is a humanized
antibody.
13. The method of claim 11, further comprising administering a
third therapeutic dose, wherein the third dose is higher than or
equal to the second dose.
14. The method of claim 11, further comprising administering a
fourth therapeutic dose, wherein the fourth dose is higher than or
equal to the third dose.
15. The method of claim 11 or claim 12, wherein administration is
intravenous or subcutaneous.
16. The method of claim 11 or claim 12, wherein administration is
not more than once per week.
17. The method of claim 11 or claim 12, wherein the antibody or
receptor binding fragment thereof is non-lymphocyte depleting.
18. The method of claim 17, wherein the antibody or receptor
binding fragment thereof is non-T-cell depleting.
19. The method of claim 11 or claim 12, wherein the first
conditioning dose is 0.7 mg/kg and the second therapeutic dose is
1.0 mg/kg.
20. The method of claim 11 or claim 12, wherein the first
conditioning dose is 0.7 mg/kg and the second therapeutic dose is
2.0 mg/kg.
Description
[0001] This is a continuation under 37 CFR .sctn.1.53(b) of
co-pending U.S. application Ser. No. 11/078,332, filed Mar. 10,
2005, which is a continuation of U.S. application Ser. No.
10/655,905, filed Sep. 5, 2003 (now abandoned), which is a
divisional of U.S. application Ser. No. 09/936,603, filed Feb. 11,
2002, now U.S. Pat. No. 6,652,855, which claims priority to
international application no. PCT/US00/07189, filed Mar. 17, 2000,
which claims the benefit of provisional patent applications Ser.
No. 60/125,351 and Ser. No. 60/125,228, both filed Mar. 19, 1999,
the disclosures of which are incorporated by reference herein in
their entirety.
FIELD OF THE INVENTION
[0002] The invention relates to methods of treating mammals, for
example humans, to reduce the occurrence of undesired
administration reactions, to treat an LFA-1 mediated disease, to
condition a mammal to tolerate high doses of a therapeutic compound
and to down modulate a cell surface receptor.
DISCUSSION OF THE BACKGROUND
[0003] Administration of many therapeutic agents rapidly induces
adverse side effects, or events, including but not limited to
fever, headache, nausea, vomiting, breathing difficulties and
changes in blood pressure. These adverse events limit the amount of
a drug or therapeutic compound that can be given, which in turn
limits the therapeutic effectiveness that could be achieved with
higher doses of the drug. There is a continuing need to develop
techniques which limit the toxicity of higher drug doses so that
therapeutic efficacy can be improved. This need exists for both
polypeptide and non-polypeptide compounds.
[0004] Antibodies are one type of polypeptide compound for which
there are frequently adverse events upon administration which limit
the dose of the compound that can be administered. One compound
associated with adverse side effects is the murine monoclonal
antibody OKT3. OKT3 binds to the CD3 protein complex that is
associated with the T cell receptor (TCR) found on the surface of
all T lymphocytes. Administration of OKT3 to humans rapidly reduces
the number of circulating T cells (e.g. OKT3 is a cell depleting
compound) and reduces the amount of cell surface TCR found on those
T cells that remain (Cosimi, et al., 1981 N Engl J Med, 305(6),
308-314). The immunosuppressive effects of OKT3 have been
therapeutically useful in the treatment of renal transplant
rejection (Goldstein & Group, 1985 M Engl J Med, 313(6),
337-342). However, administration of OKT3 induces a number of
adverse side effects, including fever, chills, nausea, vomiting and
tightness of chest. These side effects are believed to be caused by
cytokine release from T cells due to OKT3-induced activation
(Abramowicz, et al., 1989 Transplantation, 47(4), 606-608) and
complement activation (Raasveld, et al., 1993 Kidney International,
43, 1140-1149).
[0005] Several strategies have been developed to reduce the
OKT3-induced side effects. Anti-inflammatory steroids have been
shown to attenuate the OKT3-induced cytokine release (Goldman, et
al., 1989 Lancet, ii (8666), 802) (Chatenoud, et al., 1990
Transplantation, 49(4), 697-702), and indomethacin can reduce the
febrile response (First, Schroeder, Hariharan, Alexander, &
Weiskittel, 1992 Transplantation, 53 (1), 91-94). A standard 5 mg
dose of OKT3 administered as a 2 hour infusion instead of the usual
bolus injection was better tolerated and reduced complement
activation, but not the cytokine release (ten Berge, Buysmann, van
Diepen, Surachno, & Hack, 1996 Transplant Proc. 28 (6),
3217-3220). The adverse events induced by OKT3 are most significant
after the first dose. While the initial dose (typically 5 mg)
induces cytokine release and activates complement, it also
eliminates the target T cells. With fewer T cells and reduced TCR
density on those that do remain, subsequent doses of OKT3 induce
less cytokine release (Chatenoud, et al., 1989 N Engl J Med, 320
(21), 1420-1421). One group found that after four daily doses of 5
mg, dosing could safely be escalated to 10, 15, and 25 mg over the
next 3 days (Woodle, et al., 1996 Clin Transplantation, 10,
389-395).
[0006] Adverse events have also been associated with the initial
administration of monoclonal antibodies directed to other cell
surface molecules. A humanized anti-CD4 monoclonal antibody induced
fever, chills, hypotension and chest tightness when given
intravenously to psoriasis and rheumatoid arthritis patients
(Isaacs, et al., 1997 Clin Exp Immunol, 110, 158-166). This
treatment down-modulated expression of CD4 and caused a reduction
in the number of circulating CD4-positive T cells, and but was not
completely depleting. Bispecific antibodies that interact with the
CD64 molecule, a receptor for the constant region of immunoglobulin
(Fc gamma RI), and tumor associated molecules (epidermal growth
factor receptor MDX-447, or HER2/neu MDX-H210) were shown to cause
flu-like symptoms such as fever and chills after the first dose
(Curnow, 1997, Cancer Immunol Immunother, 45, 210-215). Similar to
the effect of OKT3 on T cells, these antibodies caused a decrease
in the number of circulating monocytes, which express CD64, and
stimulated increases in plasma cytokines. A single dose of another
monoclonal antibody directed to CD64 (MDX-33) down-modulated the
expression of CD64 on monocytes and also caused chills, low-grade
fever, headache and muscle aches.
[0007] The interaction of T-lymphocytes with antigen-presenting
cells (APCs) is one of the initial steps in the activation of an
immunological response to what is perceived by the immune system to
be a foreign antigen. Although much attention has been focused on
the primary interaction of the T-cell receptor with the MHC-antigen
on the APC, several other cell surface components are also involved
in T-cell activation. These ligand pairs located on the cell
surface of the T-cell and the APC include: LFA-1/ICAM-1 (also
ICAM-2 and ICAM-3), CD28/B7, CD2/LFA-3, CD4/MHC Class II, and
CD8/MHC Class I. Interfering with the binding of any of these
ligand pairs (e.g., with the use of binding molecules such as
monoclonal antibodies) may decrease, inhibit, or discontinue the
T-cell responses (de Fourgerolles et al., 1994, J. Exp. Med.,
179:619-29; Dustin, M L et al, 1986, J Immunol, 137:245-54).
[0008] LFA-1 (consisting of CD11a and CD18 subunits) interaction
with ICAM is necessary for T-cell killing, T-helper and B-cell
responses, natural killing, and antibody-dependent cytotoxicity. In
addition, LFA-1/ICAM interactions are involved in adherence of
leukocytes to endothelial cells, fibroblasts, and epithelial cells,
facilitating the migration of leukocytes from the vasculature to
the sites of inflammation (Collins, T., 1995, Science and Medicine,
28-37; Dustin, M L. et al., 1991, Annual Rev Immunology,
9:27-66).
[0009] Using antibodies that interfere with LFA-1/ICAM interactions
decreases or inhibits the inflammatory process by blocking the
activation of T-cells and/or the extravasation of leukocytes. In
vitro, monoclonal antibodies against LFA-1 or its ligands have
inhibited T-cell activation (Kuypers, T. and Roos, D., 1989,
Research in Immunology, 140:461-86; Springer, T A, 1987, Annual Rev
Immunology, 5:223-52), T-cell dependent B-cell proliferation
(Fischer, A. et al., 1986, J Immunol, 136:3198-203), target cell
lysis (Krensky, A. et al., 1983, J Immunol, 131:6711-6), and
adhesion of T-cells to vascular endothelium (Dustin, M L. et al.,
1988, Journal of Cell Biology, 107:321-31). In mice, anti-CD11a
antibodies have induced tolerance to protein antigens (Benjamin, R.
et al, 1988, European Journal of Immunology, 18:1079-88; Tanaka, Y.
et al., 1995, European Journal of Immunology, 25:1555-8), delayed
the onset and reduced the severity of experimental autoimmune
encephalomyelitis (Gordon, E J et al., 1995, Journal of
Neuroimmunology, 62:153-60), inhibited lupus-associated
autoantibody production, and prolonged survival of several types of
tissue grafts (Cavazzana-Calco M S, Sarnacki S, Haddad E, et al.,
Transplantation 1995;59(11):1576-82; Nakakura E K, McCabe S M,
Zheng B, Shorthouse R A, et al., Transplantation 1993;55(2):412-7;
Connolly M K, Kitchens E A, Chan B, et al, Clinical Immunology and
Immunopathology 1994;72(2):198-203; He Y, Mellon J, Apte R,
Niederkorn J., Investigative Ophthalmology and Visual Science
1994;35(8):3218-25; Isobe M, Yagita H, Okumura K, Ihara A., Science
1992;255:1125-7; Kato Y, Yamataka A, Yagita H, et al., Ann Surg
1996;223(1):94-100; Nishihara M, Gotoh M, Fukuzaki T, et al.,
Transplantation Proceedings 1995;27(1):372; Talento A, Nguyen M,
Blake T, et al, Transplantation 1993;55(2):418-22; van Dijken P J,
Ghayur T, Mauch P, et al., Transplantation 1990;49(5):882-6). In
human clinical studies, murine anti-CD11a monoclonal antibodies
have been shown to help prevent graft failure following bone marrow
transplantation (Cavazzana-Calco M S, Bordigoni P, Michel G, et
al., British Journal of Haematology 1996;93:131-8; Fischer A,
Friedrich W, Fasth A., Blood 1991;77(2):249-56; Stoppa A M,
Maraninchi D, Blaise D, Viens P, et al., Transplant International
1991;4:3-7) and renal transplantation (Hourmant M, Le Mauff B, Le
Meur Y, et al., Transplantation 1994;58(3):377-80; Hourmant M,
Bedrossian J, Durand D, et al., Transplantation
1996;62(11):1565-70; Le Mauff B, Hourmant M, Rougier J P, et al.,
Transplantation 1991;52(2):291-6). An immunosuppressive drug that
could reduce the incidence of both acute graft rejection and
delayed graft function, while promoting long-term survival with
minimum toxicity with the potential of tolerance induction would
provide major benefits to the field of renal transplantation.
[0010] A need continues to exist for new methods of administering
therapeutic compounds which reduces side effects and which
increases the effectiveness of the therapeutic compound.
SUMMARY OF THE INVENTION
[0011] One object of the present invention is to provide an
improved method of administering a therapeutic compound. This and
other objects which will become apparent from the following
description of enabling embodiments have been achieved by the
method of the invention.
[0012] One aspect of the invention is a method for reducing the
occurrence of fever, headache, nausea and/or vomiting associated
with administration of a therapeutic compound to a mammal in need
thereof by administering to the mammal a first conditioning dose of
a non-target cell depleting compound which binds to a cell surface
receptor on a target mammalian cell; and then administering at
least a second therapeutic dose of the compound, wherein the second
dose is higher than the first dose.
[0013] A further aspect of the invention is a method for treating
an LFA-1 mediated disorder by administering to a mammal in need
thereof a first conditioning dose of a compound which binds to
lymphocyte surface receptor LFA-1; and then administering at least
a second therapeutic dose of the compound, wherein the second dose
is higher than the first dose. LFA-1 mediated disorders
contemplated include psoriasis, asthma, rheumatoid arthritis,
multiple sclerosis and transplant rejection. In a specific
embodiment, the graft or transplant is a renal transplant.
[0014] A further aspect of the invention is a method for
conditioning a mammal to tolerate high doses of a therapeutic
compound by administering to the mammal a first conditioning dose
of a non-target cell depleting compound which binds to a cell
surface receptor on a target mammalian cell; and then administering
at least a second therapeutic dose of the compound, wherein the
second dose is higher than the first dose.
[0015] Another aspect of the invention is a method for down
modulating a cell surface receptor in a mammalian cell population
by contacting a target mammalian cell displaying a receptor
molecule on the surface thereof with a first dose of a ligand which
binds to the receptor molecule and does not deplete the mammalian
cell population; and then further contacting the mammalian cell
population with at least a second dose of the ligand, wherein the
second dose is higher than the first dose.
[0016] The following preferred embodiments apply to all the above
methods of the invention. In preferred embodiments, the therapeutic
compound comprises a polypeptide which binds to an extracellular
domain of the receptor molecule. A preferred polypeptide is an
antibody or a fragment thereof. In one embodiment, the target
mammalian cell is a lymphocyte such as a T lymphocyte. Non-T
cell-depleting compounds or antibodies that bind CD11a or CD18 cell
surface receptors on a T cell, including the humanized anti-CD11a
antibody hu1124, are specifically encompassed.
[0017] Intravenous or subcutaneous mode of administration of the
therapeutic compound is contemplated. In a specific embodiment,
administration is not more than once per week. In an additional
embodiment of each of the above methods, the methods further
comprise administering a third therapeutic dose, wherein the third
dose is higher than the second dose. Yet a further embodiment is
the administration of a fourth therapeutic dose, wherein the fourth
dose is higher than or equal to the third dose.
[0018] Also provided is composition comprising an anti-CD11a
antibody or fragment thereof, and a pharmaceutical carrier, for use
as an active pharmaceutical agent for treating an LFA-1 mediated
disorder, wherein the antibody is a non-target cell-depleting
antibody and wherein the composition is administered to the mammal
as a first conditioning dose followed by a second therapeutic dose,
wherein the second therapeutic dose is higher than the first dose.
Another aspect is the use of a compound which binds to the
lymphocyte surface receptor LFA-1 in the preparation of a
medicament for the treatment of an LFA-1 mediated disorder, which
treatment comprises administering to the mammal, a first
conditioning dose of the compound followed by a second therapeutic
dose of the compound wherein the second therapeutic dose is higher
than the first dose.
[0019] Other aspects of the invention will become apparent from the
following description of preferred embodiments which are not
intended to be limiting of the invention.
BRIEF DESCRIPTION OF THE FIGURES
[0020] FIG. 1 shows the dosing schedule used in the study of
Example 1. A indicates time of Physician's Global Assessment and
PASI, and skin biopsy.
[0021] FIG. 2 shows the changes in PASI score results by dose group
for the study of Example 1. It is a summary of Mean Percent Change
(.+-.SD) in PASI scores from baseline to Days 28 and 56. *Subject
015 withdrew consent at Day 7 and did not have any data for Days 28
and 56.
[0022] FIG. 3 shows the dose response reduction in PASI score
results for the study of Example 1.
[0023] FIGS. 4-A through 4-E show the modulation of lymphocyte
CD11a and plasma hu1124 levels for each of Groups A-E, for the
study of Example 1.
[0024] FIG. 5 summarizes histologic data from Example 1. Histologic
evidence of decreased ICAM-1, Keratin 16 (Ker 16), and CD11a
expression in psoriatic plaques is shown.
[0025] FIG. 6 shows the decrease in acute adverse events over time,
achieved in Example 1 using the method of the invention. An acute
adverse event is one or more of fever (mild), headache (mild),
nausea or vomiting within 24 hours of dosing. A single patient may
have had more than one event.
DEFINITIONS
[0026] The term "antibody" is used in the broadest sense and
specifically covers single monoclonal antibodies, antibody
compositions with polyepitopic specificity, as well as antibody
fragments (e.g., Fab, F(ab').sub.2, scFv and Fv), so long as they
exhibit the desired biological activity.
[0027] 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 except for possible naturally occurring
mutations that may be present in minor amounts. Monoclonal
antibodies are highly specific, being directed against a single
antigenic site. Furthermore, in contrast to conventional
(polyclonal) antibody preparations which 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 synthesized
by the hybridoma culture, uncontaminated by other immunoglobulins.
The modifier "monoclonal" indicates the character of the antibody
as being obtained from a substantially homogeneous population of
antibodies, and is not to be construed as requiring production of
the antibody by any particular method. For example, the monoclonal
antibodies to be used in accordance with the present invention may
be made by the hybridoma method first described by Kohler &
Milstein, Nature, 256:495 (1975), or may be made by recombinant DNA
methods (see, e.g., U.S. Pat. No. 4,816,567 (Cabilly et al.)).
[0028] 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, e.g. binding to a cell surface
receptor (U.S. Pat. No. 4,816,567 (Cabilly et al.); and Morrison et
al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).
[0029] "Humanized" forms of non-human (e.g., murine) antibodies are
chimeric immunoglobulins, immunoglobulin chains or fragments
thereof (such as Fv, Fab, Fab', F(ab').sub.2 or other
antigen-binding subsequences of antibodies) which contain minimal
sequence derived from non-human immunoglobulin. For the most part,
humanized antibodies are human immunoglobulins (recipient antibody)
in which residues from a complementarity determining region (CDR)
of the recipient are replaced by residues from a CDR of a non-human
species (donor antibody) such as mouse, rat or rabbit having the
desired specificity, affinity and capacity. In some instances, Fv
framework residues of the human immunoglobulin are replaced by
corresponding non-human residues. Furthermore, humanized antibody
may comprise residues which are found neither in the recipient
antibody nor in the imported CDR or framework sequences. These
modifications are made to further refine and optimize 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 CDR regions
correspond to those of a non-human immunoglobulin and all or
substantially all of the FR regions are those of a human
immunoglobulin consensus sequence. The humanized antibody optimally
also will comprise at least a portion of an immunoglobulin constant
region (Fc), typically that of a human immunoglobulin. For further
details see: Jones et al., Nature, 321:522-525 (1986); Reichmann et
al., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct.
Biol., 2:593-596 (1992)).
[0030] "Single-chain Fv" or "sFv" antibody fragments comprise the
V.sub.H and V.sub.L domains of antibody, wherein these domains are
present in a single polypeptide chain. Generally, the Fv
polypeptide further comprises a polypeptide linker between the
V.sub.H and V.sub.L domains which enables the sFv to form the
desired structure for antigen binding. For a review of sFv see
Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113,
Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315
(1994).
[0031] The term "diabodies" refers to small antibody fragments with
two antigen-binding sites, which fragments comprise a heavy chain
variable domain (V.sub.H) connected to a light chain variable
domain (V.sub.L) in the same polypeptide chain (V.sub.H and
V.sub.L). By using a linker that is too short to allow pairing
between the two domains on the same chain, the domains are forced
to pair with the complementary domains of another chain and create
two antigen-binding sites. Diabodies are described more fully in,
for example, EP 404,097; WO 93/11161; and Hollinger et al., Proc.
Natl. Acad. Sci. USA 90:6444-6448 (1993).
[0032] The expression "linear antibodies" when used throughout this
application refers to the antibodies described in Zapata et al.
Protein Eng. 8(10):1057-1062 (1995). Briefly, these antibodies
comprise a pair of tandem Fd segments
(V.sub.H-C.sub.H1-V.sub.H-C.sub.H1) which form a pair of antigen
binding regions. Linear antibodies can be bispecific or
monospecific.
[0033] A "variant" antibody, refers herein to a molecule which
differs in amino acid sequence from a "parent" antibody amino acid
sequence by virtue of addition, deletion and/or substitution of one
or more amino acid residue(s) in the parent antibody sequence. In
the preferred embodiment, the variant comprises one or more amino
acid substitution(s) in one or more hypervariable region(s) of the
parent antibody. For example, the variant may comprise at least
one, e.g. from about one to about ten, and preferably from about
two to about five, substitutions in one or more hypervariable
regions of the parent antibody. Ordinarily, the variant will have
an amino acid sequence having at least 75% amino acid sequence
identity with the parent antibody heavy or light chain variable
domain sequences, more preferably at least 80%, more preferably at
least 85%, more preferably at least 90%, and most preferably at
least 95%. Identity or homology with respect to this sequence is
defined herein as the percentage of amino acid residues in the
candidate sequence that are identical with the parent antibody
residues, after aligning the sequences and introducing gaps, if
necessary, to achieve the maximum percent sequence identity. None
of N-terminal, C-terminal, or internal extensions, deletions, or
insertions into the antibody sequence shall be construed as
affecting sequence identity or homology. The variant retains the
ability to bind the receptor and preferably has properties which
are superior to those of the parent antibody. For example, the
variant may have a stronger binding affinity, enhanced ability to
activate the receptor, etc. To analyze such properties, one should
compare a Fab form of the variant to a Fab form of the parent
antibody or a full length form of the variant to a full length form
of the parent antibody, for example, since it has been found that
the format of the antibody impacts its activity in the biological
activity assays disclosed herein. The variant antibody of
particular interest herein is one which displays at least about 10
fold, preferably at least about 20 fold, and most preferably at
least about 50 fold, enhancement in biological activity when
compared to the parent antibody.
[0034] The term "down modulating a cell surface receptor" means a
process or method which reduces the number of molecules of the
receptor on the surface of a cell type relative to the number of
molecules of the receptor before the process or method was
performed. For example, the administration of a therapeutic
compound which binds to a cell surface receptor will down regulate
the receptor if the number of receptor molecules on the surface of
the cell is less after administration than before administration of
the compound. The number of cell surface receptor molecules can be
measured histologically using known staining and counting
methods.
[0035] A "conditioning dose" is a dose which attenuates or reduces
the frequency or the severity of first dose adverse side effects
associated with administration of a therapeutic compound. The
conditioning dose may be a therapeutic dose, a sub-therapeutic
dose, a symptomatic dose or a sub-symptomatic dose. A therapeutic
dose is a dose which exhibits a therapeutic effect on the patient
and a sub-therapeutic dose is a dose which does not exhibit a
therapeutic effect on the patient treated. A symptomatic dose is a
dose which induces at least one adverse effect on administration
and a sub-symptomatic dose is a dose which does not induce an
adverse effect.
[0036] The term "cell surface receptor" as used herein means any
molecule displayed on the surface of a cell and available for
binding by therapeutic compounds which contact the surface of the
cell. Such a cell surface molecule is a "receptor" for the
therapeutic compound. Human leukocyte surface markers, including
but not limited to CD2, CD29, CD40, CD49a-d and CD58 may be cell
surface receptors within the invention. Suitable cell surface
receptors also include cell adhesion molecules such as the
leukocyte integrins CD11a/CD18, CD11b/CD18, CD11c/CD18 and
CD11d/CD18 which are heterodimeric surface receptor molecules. A
therapeutic compound may bind to either member of the heterodimeric
pair. The cell surface receptor may have an extracellular domain
which available for binding to a therapeutic compound as well as
transmembrane and intracellular domains. Other examples of
receptors include protein kinase receptors capable of intracellular
signaling through tyrosine phosphorylation, etc.
[0037] The term "non-target cell depleting compound" or a compound
or ligand which "does not deplete" a cell population means a
compound which binds to a cell surface receptor molecule, but does
not substantially reduce the number of cells in the cell
population. A non-depleting compound, for example, will reduce the
number of cells in a cell population by about 50% or less,
preferably by about 30% or less, more preferably by about 20% or
less, relative to the number of target cells in the cell population
prior to contact or treatment with the compound.
[0038] The term "LFA-1-mediated disorders" refers to pathological
states caused by cell adherence interactions involving the LFA-1
receptor on lymphocytes. Examples of such disorders include T cell
inflammatory responses such as inflammatory skin diseases including
psoriasis; responses associated with inflammatory bowel disease
(such as Crohn's disease and ulcerative colitis); adult respiratory
distress syndrome; dermatitis; meningitis; encephalitis; uveitic;
allergic conditions such as eczema and asthma and other conditions
involving infiltration of T cells and chronic inflammatory
responses; skin hypersensitivity reactions (including poison ivy
and poison oak); atherosclerosis; leukocyte adhesion deficiency;
autoimmune diseases such as rheumatoid arthritis, systemic lupus
erythematosus (SLE), diabetes mellitus, multiple sclerosis,
Reynaud's syndrome, autoimmune thyroiditis, experimental autoimmune
encephalomyelitis, Sjorgen's syndrome, juvenile onset diabetes, and
immune responses associated with delayed hypersensitivity mediated
by cytokines and T-lymphocytes typically found in tuberculosis,
sarcoidosis, polymyositis, granulomatosis and vasculitis;
pernicious anemia; diseases involving leukocyte diapedesis; CNS
inflammatory disorder, multiple organ injury syndrome secondary to
septicaemia or trauma; autoimmune haemolytic anemia; myethemia
gravis; antigen-antibody complex mediated diseases; all types of
transplantation rejection, including graft vs. host or host vs.
graft disease; etc.
[0039] "Treating" a disease, disorder, condition or cell population
includes therapy and prophylactic treatment on an acute short term
basis and on a chronic long-term basis.
[0040] The term "mammal" refers to any animal classified as a
mammal, including humans, domestic and farm animals, and zoo,
sports, or pet animals, such as dogs, horses, cats, cows, etc.
Preferably, the mammal herein is human.
[0041] A "ligand" or a "compound" which binds to a receptor
molecule on the surface of a cell may be any compound capable of
binding to the receptor molecule. The therapeutic compound may, for
example, be a small organic molecule (preferably having a molecular
weight of about 1000 g/mole or less) or a polypeptide. Suitable
polypeptide compounds or ligands can be prepared by methods known
in the art, for example, by isolating peptides or proteins having
high binding affinities for the surface molecule using phage
display technology. Phage display methods are disclosed, for
example, in U.S. Pat. No. 5,750,373; U.S. Pat. No. 5,821,047; U.S.
Pat. No. 5,780,279; U.S. Pat. No. 5,403,484; U.S. Pat. No.
5,223,409; U.S. Pat. No. 5,571,698; etc. In these methods, large
libraries of peptides or proteins displayed on the surface of phage
are produced and screened or panned to select those members of the
library which bind strongly to a target molecule which is generally
immobilized on a solid support. These methods may be used to select
peptides or proteins which bind to a cell surface receptor by
screening a phage display library using the cell surface receptor
as the target molecule. Repeated rounds of selection and separation
of the binders having high binding affinity produces polypeptide
compounds capable of binding to the cell surface receptor. The
polypeptide may be an antibody or an antibody fragment.
[0042] A "therapeutic" compound is any compound which is used in
treating a mammal.
[0043] The term "a compound which binds to lymphocyte surface
receptor LFA-1" generally refers to any compound capable of binding
to either component of LFA-1. The compound may be a protein which
recognizes and binds to LFA-1, for example a binding protein, an
antibody directed against either CD11a or CD18 or both, but also
includes ICAM-1, soluble forms of ICAM-1 (e.g., the ICAM-1
extracellular domain, alone or fused to an immunoglobulin
sequence), antibodies to ICAM-1, and fragments thereof, or other
molecules capable of inhibiting the interaction of LFA-1 and
ICAM-1.
[0044] The term "anti-LFA-1 antibody" or "anti-LFA-1 MAb" refers to
an antibody directed against either CD11a or CD18 or both. The
anti-CD11a antibodies include, e.g., MHM24 [Hildreth et al., Eur.
J. Immunol., 13: 202-208 (1983)], R3.1 (IgG1) [R. Rothlein,
Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Conn.],
25-3 (or 25.3), an IgG1 available from Immunotech, France [Olive et
al., in Feldmann, ed., Human T cell Clones, A new Approach to
Immune Regulation, Clifton, N.J., Humana, 1986 p. 173], KBA (IgG2a)
[Nishimura et al., Cell. Immunol., 107: 32 (1987); Nishimura et
al., ibid., 94: 122 (1985)], M7/15 (IgG2b) [Springer et al.,
Immunol. Rev., 68: 171 (1982)], IOT16 [Vermot Desroches et al.,
Scand. J. Immunol., 33: 277-286 (1991)], SPVL7 [Vermot Desroches et
al., supra], and M17 (IgG2a), available from ATCC, which are rat
anti-murine CD11a antibodies. Preferred anti-CD11a antibodies are
the humanized antibodies described in WO 98/23761.
[0045] Examples of anti-CD18 antibodies include MHM23 [Hildreth et
al., supra], M18/2 (IgG2a) [Sanches-Madrid et al., J. Exp. Med.,
158: 586 (1983)], H52 [Fekete et al., J. Clin. Lab Immunol., 31:
145-149 (1990)], Mas191c [Vermot Desroches et al., supra], IOT18
[Vermot Desroches et al., supra], 60.3 [Taylor et al., Clin. Exp.
Immunol., 71: 324-328 (1988)], and 60.1 [Campana et al., Eur. J.
Immunol., 16: 537-542 (1986)].
[0046] Other examples of suitable LFA-1 binding molecules,
including antibodies, are described in Hutchings et al., supra, WO
91/18011 published Nov. 28, 1991, WO 91/16928 published Nov. 14,
1991, WO 91/16927 published Nov. 14, 1991, Can. Pat. Appln.
2,008,368 published Jun. 13, 1991, WO 90/15076 published Dec. 13,
1990, WO 90/10652 published Sep. 20, 1990, EP 387,668 published
Sep. 19, 1990, EP 379,904 published Aug. 1, 1990, EP 346,078
published Dec. 13, 1989, U.S. Pat. No. 5,071,964, U.S. Pat. No.
5,002,869, Australian Pat. Appln. 8815518 published Nov. 10, 1988,
EP 289,949 published Nov. 9, 1988, and EP 303,692 published Feb.
22, 1989.
[0047] The antibody is appropriately from any source, including
chicken and mammalian such as rodent, goat, primate, and human.
Preferably, the antibody is from the same species as the species to
be treated, and more preferably the antibody is human or humanized
and the host is human. While the antibody can be a polyclonal or
monoclonal antibody, preferably it is a monoclonal antibody, which
can be prepared by conventional technology. The antibody is an
IgG-1, -2, -3, or -4, IgE, IgA, IgM, IgD, or an intraclass chimera
in which Fv or a CDR from one class is substituted into another
class. The antibody may have an Fc domain capable of an effector
function or may not be capable of binding complement or
participating in ADCC.
Detailed Description of the Preferred Embodiments
[0048] It has now been discovered that a dosing schedule in which a
first conditioning dose of a non-target cell depleting compound
which binds to a cell surface receptor on a target mammalian cell
is followed by a second higher dose of the compound, is effective
in conditioning a mammal to tolerate increasing or higher doses of
the therapeutic compound. This dosing schedule allows one to reduce
the occurrence of adverse effects which arise from the initial
administration and subsequent administrations of the therapeutic
compound, that is, the method of the invention reduces the first
dose adverse effects of administration and condition the mammal to
further higher doses. Although some adverse effects such as fever,
headache, nausea, vomiting, breathing difficulties, myalgia, chills
and changes in blood pressure will still be observed, the frequency
and/or severity of these adverse effects is surprisingly reduced
relative to administration using conventional dosing schedules such
as daily administration of equal doses of a therapeutic compound.
The method of the present invention, therefore, allows one to
increase subsequent doses and obtain the therapeutic benefit of
higher doses of the therapeutic compounds, while, at the same time,
minimizing the occurrence of adverse side effects.
[0049] The therapeutic compound which is administered using the
dosing schedule of the present invention is a non-target cell
depleting compound which binds to a cell surface receptor on the
target cell. Such a compound does not substantially reduce the
number of cells in the cell population. For example, if the target
cell is a lymphocyte, administration of a non-lymphocyte cell
depleting compound according to the dosing schedule of the present
invention will result in binding of the compound to a lymphocyte
cell surface receptor, but will not result in a decrease in the
number of circulating lymphocytes over the course of administration
of the therapeutic compound. It is contemplated that administration
of the therapeutic compound will, in some cases, result in
increases in the number of target cells in the cell population,
however, and such an effect is to be considered within the scope of
the present invention.
[0050] The non-target cell depleting compound which is administered
according to the method of the invention may be any non-depleting
therapeutic compound which is capable of binding to a cell surface
receptor. Many therapeutic compounds are well known to exert a
therapeutic effect by binding to a selective cell surface marker or
receptor. These known therapeutic compounds will be apparent to one
having ordinary skill in the art and may be used in the method of
the present invention. Suitable therapeutic compounds include
non-peptidic organic compounds, preferably having a molecular
weight less than about 1,000 g/mol, more preferably less than about
600 g/mol; peptide therapeutic compounds, generally containing 8 to
about 200, preferably about 15 to about 150, more preferably about
20 to about 100 amino acid residues; and protein therapeutic
compounds, generally having secondary, tertiary and possibly
quaternary structure. Suitable peptides compounds can be prepared
by known solid-phase synthesis or recombinant DNA technology which
are well known in the art.
[0051] A particularly preferred method of selecting a non-depleting
peptide compound is through the use of phage display technology.
Using known phage display methods, libraries of peptides or
proteins are prepared in which one or more copies of individual
peptides or proteins are displayed on the surface of a
bacteriophage particle. DNA encoding the particular peptide or
protein is within the phage particle. The surface-displayed
peptides or proteins are available for interaction and binding to
target molecules which are generally immobilized on a solid support
such as a 96-well plate or chromatography column support material.
Binding and/or interaction of the display peptide or protein with a
target molecule under selected screening conditions allows one to
select members of the library which bind or react with the target
molecule under the selected conditions. For example, peptides which
bind under particular pH or ionic conditions may be selected.
Alternatively, a target cell population can be immobilized on a
solid surface using known techniques and the peptide or protein
phage library can be panned against the immobilized cells to select
peptides or proteins which bind to cell surface receptors on the
target cell population. Phage display techniques are disclosed, for
example, in U.S. Pat. No. 5,750,373; U.S. Pat. No. 5,821,047; U.S.
Pat. No. 5,780,279; U.S. Pat. No. 5,403,484; U.S. Pat. No.
5,223,407; U.S. Pat. No. 5,571,698; and others.
[0052] One category of preferred polypeptide non-depleting
compounds, are compounds containing an antibody or a fragment
thereof which immunologically recognize and bind to cell surface
receptors. Methods of preparing antibodies to specific cell surface
receptors are well known in the art and have been practiced for
many years. Suitable antibodies may be prepared using conventional
hybridoma technology or by recombinant DNA methods. Preferred
antibodies are humanized forms of non-human antibodies.
Alternatively, antibodies may be prepared from antibody phage
libraries using methods described, for example, in U.S. Pat. No.
5,565,332; U.S. Pat. No. 5,837,242; U.S. Pat. No. 5,858,657; U.S.
Pat. No. 5,871907; U.S. Pat. No. 5,872,215; U.S. Pat. No.
5,733,743, and others. Suitable compounds include full-length
antibodies as well as antibody fragments such as Fv, Fab, Fab' and
F (ab').sub.2 fragments which can be prepared by reformatting the
full length antibodies using known methods.
[0053] Additional preferred polypeptide therapeutic compounds are
immunoadhesin molecules also known as hybrid immunoglobulins. These
polypeptides are useful as cell adhesion molecules and ligands and
also useful in therapeutic or diagnostic compositions and methods.
An immunoadhesin typically contains an amino acid sequence of a
ligand binding partner protein fused at its C-terminus to the
N-terminus of an immunoglobulin constant region sequence. A
suitable immunoadhesin may contain the extracellular domain of a
leukocyte integrin molecule, e.g. LFA-1, LFA-2, LFA-3, MAC-1,
p150,95, aDb2, etc., fused to the hinge and CH2 and/or CH3
sequences of the a human IgG constant region. Immunoadhesins and
methods of preparing the same are described in U.S. Pat. No.
5,428,130; U.S. Pat. No. 5,714,147; U.S. Pat. No. 4,428,130; U.S.
Pat. No. 5,225,538; U.S. Pat. No. 5,116,964; U.S. Pat. No.
5,098,833; U.S. Pat. No. 5,336,603; U.S. Pat. No. 5,565,335;
etc.
[0054] In a method of the invention, adverse side effects are
reduced by administering to a mammal a first conditioning dose of a
non-target cell-depleting compound which binds to a cell surface
receptor on a target mammalian cell followed by a therapeutic dose.
The term "therapeutic" in this context means that the compounds
binds to the surface of the target cell and produce a change in the
symptoms or conditions associated with the disease or condition
which is being treated. It is sufficient that a therapeutic dose
produce an incremental change in the symptoms or conditions
associated with the disease; a cure or complete remission of
symptoms is not required. One having ordinary skill in this art can
easily determine whether a dose is therapeutic by establishing
criteria for measuring changes in symptoms or conditions of the
disease being treated and then monitoring changes in these criteria
according to known methods. External physical conditions,
histologic examination of affected tissues in patients or the
presence or absence of specific cells or compounds, including in a
lesion, associated with a disease may provide objective criteria
for evaluating therapeutic effect. In one example, the method of
the invention may be used to treat psoriasis where therapeutic
effect is determined by a physician's global assessment (PGA) of
the patient and by Psoriases Area and Severity Index (PASI) scores.
A decrease in PASI score indicates a therapeutic effect. Psoriatic
disease activity can also be determined based on Overall Lesion
Severity (OLS) scale, percentage of total body surface area (BSA)
affected by psoriasis, and psoriasis plaque thickness. For asthma,
one indicator of therapeutic effect is a decrease in nonspecific
airway hyperresponsiveness to methacholine challenges (basal and
post-allergen; see Examples), upon treatment by the method of the
invention. Airway hyperresponsiveness can be measured by FEV.sub.1
(volume of air that can be forced from the lungs in 1 second). For
transplant or graft survival and function, therapeutic
effectiveness can be measured, e.g., by the incidence of acute
graft rejection, by graft function, and length of graft survival,
as described in the Examples. Other indicators of therapeutic
effect will be readily apparent to one having ordinary skill in the
art and may be used to establish efficacy of the dose.
[0055] In the method of the invention, the first dose in followed
by a second dose which is higher than the first dose, that is,
contains a greater amount of the therapeutic compound. The first
dose serves to condition the mammal to tolerate the higher second
therapeutic dose. In this way, the mammal is able to tolerate
higher doses of the therapeutic compound than could be administered
initially. Also within the scope of the present invention are
additional doses, which may be administered after the second dose.
For example, an additional, third dose which is higher than or
equal to the second dose and an additional fourth dose which is
higher than or equal to the second or third dose are contemplated
within the method of present invention.
[0056] In a further embodiment, the first dose may be repeated one
or more times before the second higher dose is administered. The
first dose may be administered, for example, one, two or three
times, most preferably only one time before the second higher dose
is administered.
[0057] The doses may be administered according to any time schedule
which is appropriate for treatment of the disease or condition. For
example, the dosages may be administered on a daily, weekly,
biweekly or monthly basis in order to achieve the desired
therapeutic effect and reduction in adverse effects. The dosages
can be administered before, during or after the development of the
disorder. For example, to prevent host versus graft or graft versus
host rejection, the initial conditioning dose may be administered
before, during or after transplantation has occurred. The specific
time schedule can be readily determined by a physician having
ordinary skill in administering the therapeutic compound by routine
adjustments of the dosing schedule within the method of the present
invention. The time of administration of the first and second
dosages as well as subsequent dosages will be adjusted to minimize
adverse effects while maintaining a maximum therapeutic effect. The
occurrence of adverse effects can be monitored by routine patient
interviews and adjusted to minimize the occurrence of side effects,
in particular, fever, headache, nausea and/or vomiting by adjusting
the time of the dosing. Any dosing time is to be considered to be
within the scope of the present invention so long as the first
conditioning dose of the non-depleting compound is administered
followed by a second higher dose of the compound. For example,
additional doses may be on a daily or weekly schedule followed by
subsequent biweekly or monthly doses.
[0058] The dosage amount will depend on the specific disease or
condition which is treated and can be readily determined using
known dosage adjustment techniques by a physician having ordinary
skill in treatment of the disease or condition. The dosage amount
will generally lie with an established therapeutic window for the
therapeutic compound which will provide a therapeutic effect while
minimizing additional morbidity and mortality. Typically,
therapeutic compounds will be administered in a dosage ranging from
0.001 mg/kg to about 100 mg/kg per dose, preferably 0.1-20 mg/kg.
The preferred dose of about 0.1-20 mg/kg is particularly useful for
non-cell depleting compounds containing antibodies or fragments
thereof.
[0059] Typically, the therapeutic compound used in the method of
this invention is formulated by mixing it at ambient temperature at
the appropriate pH, and at the desired degree of purity, with
physiologically acceptable carriers, i.e., carriers that are
non-toxic to recipients at the dosages and concentrations employed.
The pH of the formulation depends mainly on the particular use and
the concentration of antagonist, but preferably ranges anywhere
from about 3 to about 8. Formulation in an acetate buffer at pH 5
is a suitable embodiment. Where the therapeutic compound is an
anti-LFA-1 antibody (such as hu1124), a suitable embodiment is a
formulation at pH 6.0.
[0060] The therapeutic compound, e.g. an anti-LFA-1 antibody, for
use herein is preferably sterile. Sterility can be readily
accomplished by sterile filtration through (0.2 micron) membranes.
Preferably, therapeutic peptides and proteins are stored as aqueous
solutions, although lyophilized formulations for reconstitution are
acceptable.
[0061] The therapeutic compound may be formulated, dosed, and
administered in a fashion consistent with good medical practice.
Factors for consideration in this context include the particular
disorder being treated, the particular mammal being treated, the
clinical condition of the individual patient, the cause of the
disorder, the site of delivery of the agent, the method of
administration, the time scheduling of administration, and other
factors known to medical practitioners. The "therapeutically
effective amount" of the therapeutic compound to be administered
will be governed by such considerations, and is the minimum amount
necessary to prevent, ameliorate, or treat the disease, for example
an LFA-1-mediated disorder, including treating rheumatoid
arthritis, psoriasis, multiple sclerosis, asthma, or prolonging
survival of a transplanted graft. Such amount is preferably below
the amount that is toxic to the host or renders the host
significantly more susceptible to infections.
[0062] The therapeutic compound may be administered by any suitable
means, including parenteral, subcutaneous, intraperitoneal,
intrapulmonary, intranasal, and intralesional administration.
Parenteral infusions include intramuscular, intravenous,
intraarterial, intraperitoneal, or subcutaneous administration.
Preferably the dosing is given by injections, most preferably
intravenous or subcutaneous injections, depending in part on
whether the administration is brief or chronic.
[0063] In another aspect of the invention, a method of
down-modulating a cell surface receptor in a cell population in a
mammal is provided by contacting a target mammalian cell displaying
a receptor molecule on the surface thereof with a first dose of a
ligand which binds to the receptor molecule and does not deplete
the mammalian cell population; and then further contacting the
mammalian cell population with a second dose of the ligand, wherein
the second dose is higher than the first dose. Binding of the
ligand to the cell surface receptor reduces the number of receptors
on the surface of the cell even though the numbers of cells are not
substantially reduced. This reduction in surface receptors is
associated with reduced side effects such as reduced fever,
headache, nausea and/or vomiting.
[0064] The non-cell depleting compound or ligand used in this
method may be same as described above, and may be provided to the
mammal in the same manner of administration, using the same dosing
schedule and in the same dosage amounts as described above.
[0065] Another aspect of the invention is the treatment of an LFA-1
mediated disorder by administering to a mammal, preferably a human
patient, in need of such a treatment a first conditioning dose of a
compound which binds to lymphocyte surface receptor LFA-1; and
administering a second therapeutic dose of the compound, where the
second dose is higher than the first dose. This aspect of the
invention is different than conventional dosing methods for the
treatment of such diseases which generally treat with regularly
spaced, even doses of a therapeutic compound. For example, the
immunoadhesin LFA-3TIP, which is a recombinant dimeric protein
consisting of the first extracellular domain of human LFA-3 fused
to the hinge and CH2 and CH3 human IgG regions, has been
administered in equal weekly doses of 0.005 mg/kg, 0.025 mg.kg,
0.050 mg/kg or 0.075 mg/kg. In a second study, patients received
0.05, 0.10 or 0.15 mg/kg in equal doses every four weeks (Krueger
et al.). In contrast, the method of the invention provides a first
conditioning dose and then a second therapeutic higher dose.
[0066] This method of dose scheduling conditions the patient to
tolerate higher doses of the therapeutic compound than would be
tolerated by the patient, particularly when there are adverse
effects on the patients due to administration of the therapeutic
compound. The low first dose of the method of the invention is
useful for reducing the fever, headache, nausea, vomiting, etc.
which often accompany an initial administration of a drug compound.
However, it is possible to give a low first dose, within the scope
of the invention, to patients who do not experience adverse effects
on first administration.
[0067] The first dose conditions the patient to receive a second
higher dose with a reduction in adverse effects which may be
observed with higher doses of therapeutic compounds. Generally, as
the dosage amount increases, the number of adverse effects also
increases. The method of the invention allows administration of
larger therapeutic doses more quickly and with fewer adverse
effects. This improves the effectiveness of the therapy since the
patient is able to tolerate larger doses and for a longer time
since there are fewer unpleasant side effects.
[0068] Any compound which binds to a lymphocyte surface receptor
LFA-1 and reduces the severity of symptoms or conditions associated
with an LFA-1 mediated disease may be used in this embodiment of
the invention. Preferred compounds are peptide or protein
compounds, more preferably such compounds which are or which
contain an antibody or fragment thereof or which are fusions to an
antibody fragment such as an immunoadhesin. Particularly preferred
compounds are anti-CD11a antibodies or compounds containing
fragments thereof.
[0069] As described above, the dosage amount will depend on the
specific LFA-1 mediated disease which is treated and can be readily
determined using known dosage adjustment techniques by a physician
having ordinary skill in treatment of these diseases. The dosage
amount will generally lie within an established therapeutic window
for the therapeutic compound which will provide a therapeutic
effect while minimizing additional morbidity and mortality.
Typically, therapeutic compounds will be administered in a dosage
ranging from 0.001 mg/kg to about 100 mg/kg per dose, preferably
0.1-20 mg/kg. The preferred dose of about is particularly useful
for compounds containing antibodies or fragments thereof.
[0070] The therapeutic compound for treatment of an LFA-1 mediated
disease may be administered by any suitable means, including
parenteral, subcutaneous, intraperitoneal, intrapulmonary,
intranasal, and intralesional administration. Parenteral infusions
include intramuscular, intravenous, intraarterial, intraperitoneal,
or subcutaneous administration. Preferably the dosing is given by
injections, most preferably intravenous or subcutaneous injections,
depending in part on whether the administration is brief or
chronic. As described above in more detail, the therapeutic
compound for treatment of an LFA-1 mediated disease may be
formulated, dosed, and administered in a fashion consistent with
good medical practice.
[0071] Psoriasis is an inflammatory disease characterized by
hyperproliferation of keratinocytes and accumulation of activated T
cells in the epidermis and dermis of psoriatic lesions. The
upregulation of ICAM-1 on keratinocytes and its interaction with
T-cell LFA-1 in lesional skin indicate that treatment with an
anti-CD11a antibody might interfere with the disease process in
psoriasis.
[0072] Allergic asthma is characterized by the cardinal features of
airway inflammation, reversible airway obstruction, and
hyperresponsiveness. In people with allergic asthma, lymphocytes
are believed to play a central role in the asthmatic inflammatory
response to an inhaled allergen. Chronic asthma symptoms may result
from continual activation of lung lymphocytes in response to
chronic exposure to perennial allergens (e.g., house dust mite, dog
dander, or cat hair) or from sequential exposure to seasonal
allergens to which the patient is reactive.
[0073] Within minutes of an inhaled exposure to an aerosolized
allergen, the patient with allergic asthma will experience an
immediate response (the early asthmatic response or EAR), which is
characterized by a fall in the volume of air that can be forced
from the lungs in 1 second (FEV.sub.1) over 0-2 hours. In most
cases, this fall in FEV.sub.1 is reversible by treatment with a
.beta.-agonist bronchodilator. Approximately 50% of these patients
will go on to experience a second fall in FEV.sub.1 (the late
asthmatic response or LAR) 3-7 hours after the initial allergen
exposure. LAR is associated with more pronounced airway
inflammation and increased bronchial reactivity to nonspecific
stimuli. Airway hyperresponsiveness to the non-allergen-specific
challenge of methacholine is another cardinal feature of asthma.
Hyperresponsiveness of the airways in response to low levels of
methacholine is recognized by a decrease in FEV.sub.1 and may be
exacerbated by exposure of the airways to allergen, viral
infection, or physical irritants. Aerosolized allergen bronchial
challenge, or bronchoprovocation, can be performed in the
laboratory in patients with allergic asthma and is useful and
relevant as a model for the study of anti-asthma medications
(Crescioli et al. 1991 Ann Allergy 66:245-51; Cockcroft et al.,
1987, Am Rev Respir Dis 135:264-267; Ward et al. 1993, Am Rev
Respir Dis 147:518-523.). Most drugs with proven efficacy in asthma
management attenuate airway responses (EAR and/or LAR) to inhaled
allergens administered in allergen bronchial challenges.
[0074] Interleukins (IL-5, IL-3) and granulocyte/macrophage colony
stimulating factor are known to be important for eosinophil
differentiation, maturation, adherence, activation, and
degranulation. These cytokines are produced by T cells isolated
from patients with asthma (Walker et al., 1991, J. Immunol.
146:1829-35). The T-cell products IL-4 and IL-13 are key mediators
of inflammation, increasing IgE levels. Activated T cells are
increased in peripheral blood, bronchoalveolar lavage, and
bronchial biopsy specimens from patients with asthma (Azzawi et al.
1990, Am Rev Resp Dis 142:1407-13; Corrigan & Kay 1990, Am Rev
Respir Dis 141:970-7). Lymphocytes are the predominant cell type
identified in morphometric studies of submucosal biopsies obtained
from patients with asthma (Djukanovic et al., 1992, Am Rev Respir
Dis 145:669-74).
EXAMPLES
[0075] The method of the invention is exemplified by the treatment
of psoriasis, asthma and kidney transplant rejection (LFA-1
mediated diseases) with a therapeutic compound (an antibody) which
binds to a cell surface receptor (CD11a). These examples also
provide a representative example of down modulating a cell surface
receptor in a mammalian cell population, reducing side effects and
of conditioning a mammal to tolerate high doses of a therapeutic
compound.
Example 1
Phase I Multiple-Dose, Dose-Escalation Study of the Effects of
hu1124 in Subjects with Moderate to Severe Psoriases. (HUPS249)
[0076] The study drug, hu1124, is a known humanized anti-CD11a
antibody [see WO 98/23761 (humanized MHM24(Fab)-8); Werther W A, et
al., Humanization of an anti lymphocyte function-associated antigen
(LFA)-1 monoclonal antibody and re-engineering of the humanized
antibody for binding to rhesus LFA-1. J Immunol 1996;157:4986-95).
hu1124 is a humanized IgG1 version of a murine anti-CD11a
monoclonal antibody, MHM24, which recognizes human and chimpanzee
CD11a. Humanization of MHM24 was accomplished by grafting the
murine complementarity determining regions (hypervariable region)
into consensus human IgG1/.kappa. heavy- and light-chain sequences.
For H and L chain V sequences of hu1124, refer to sequences shown
in FIG. 1 in Werther et al., 1996, and in FIGS. 1A and 1B in WO
98/23761, incorporated herein by reference.
[0077] The study drug, hu1124, was supplied as a single-use, clear,
colorless, sterile, non-pyrogenic solution in a glass vial. Each
vial contained 10 mL of solution at a concentration of 4 mg/mL in
10 mM sodium acetate pH 5.0, with 0.02% polysorbate 20, 0.1% sodium
acetate trihydrate, and 4% mannitol. No preservative was added to
the solution. All study drug was stored at 2-8.degree. C.
(35.6-46.4.degree. F.). The study drug was administered to subjects
by continuous intravenous infusion over 90 minutes into a
peripheral vein. The amount of drug to be given was based on
subject weight and the dosage group to which the subject was
assigned.
[0078] Safety was assessed by adverse events, clinical laboratory
assessments, and pre- and post-treatment vital signs. Immunological
activity was monitored by testing for the effects on cell-mediated
immunity reactions (delayed hypersensitivity), tetanus antibody
responses, and lymphocyte subpopulations (flow cytometry). Efficacy
was monitored by changes in clinical signs and symptoms of the
disease (including Psoriasis Area and Severity Index, PASI, scores)
and global changes compared with the baseline condition. Skin
biopsies were studied for the effects of hu1124 on lymphocytes
within psoriatic lesions. Pharmacokinetics of the study drug were
assessed by serial monitoring of plasma samples for hu1124
throughout the 98 days following the start of the study.
[0079] Thirty-nine subjects with moderate to severe plaque
psoriasis were enrolled at eight study centers. The subjects
included Caucasian, Black, Asian and Hispanic subjects. The
subjects ranged in age from 26 to 73 years, with only one subject
older than 70. Subject weight ranged from 65 to 122 kg. Between 15
to 72%, overall median of 33%, of the body surface area was
psoriatic for the subjects enrolled in this study. Baseline PASI
scores ranged from 15 to 42, with an overall median of 23 for the
39 subjects in this study. Each of the 39 subjects received
multiple doses of hu1124, ranging from 0.1-1.0 mg/kg, administered
by intravenous infusion. Of the 39 subjects, 10 were enrolled in
one of the 0.1 mg/kg dose groups (four in the 0.1 mg/kg
administered every two weeks dose group and six in the 0.1 mg/kg/wk
dose group), 17 were enrolled in the 0.3 mg/kg/wk dose group, six
were enrolled in the 0.3-0.6 mg/kg/wk dose group, and six were
enrolled in the 0.3-1.0 mg/kg/wk dose group. All but five subjects
completed the study. All subjects were evaluated for safety and
efficacy. The table below shows the treatment schedule and number
of patients in each arm of the study. TABLE-US-00001 TABLE 1 Dose
Escalation (mg/kg) and Treatment Schedule Day Day Day Day Day n Day
0 Day 7 14 21 28 35 42 Group A 4 0.1 -- 0.1 -- 0.1 -- 0.1 Group B 6
0.1 0.1 0.1 0.1 0.1 0.1 0.1 Group C 17 0.3 0.3 0.3 0.3 0.3 0.3 0.3
Group D 6 0.3 0.4 0.6 0.6 0.6 0.6 0.6 Group E 6 0.3 0.4 0.6 1.0 1.0
1.0 1.0
Results--Safety:
[0080] The first-dose acute adverse events were primarily fever
(reported by 17/39 or 44% of subjects), headache (reported by 7/39
or 18% of subjects), nausea (reported by 5/39 or 13% of subjects),
and vomiting (2/39 or 5% of subjects). Acute (within 48 hours after
dosing) adverse events of fever, headache, or vomiting after the
first dose were not reported in subjects who received 0.1 mg/kg
every other week or every week. The majority of acute adverse
events were mild in severity. Importantly, the frequency of acute
adverse events decreased with subsequent doses in each dose
group.
[0081] Multiple infusions of hu1124 were safe and well-tolerated by
the subjects in this study. The overall decreasing incidence of
acute adverse events after the first dose in each dose group
indicates that the acute adverse events are the result of initial
drug exposure and not absolute drug level, which in turn indicates
an adaptation or conditioning response to the "desensitizing" or
"conditioning" dosing schedule of the invention.
Results--Pharmacokinetic/Pharmacodynamics
[0082] The mean peak and trough plasma levels of hu1124 appeared to
be dose dependent and increased as the dose level of study
medication increased. No accumulation of hu1124 was observed in the
lower dose groups and a small degree of accumulation was observed
after the maximum doses were infused in the higher dose groups. A
decrease in CD11a expression was observed within 2-4 hours after
study medication administration in all dose groups, with full
recovery noted before the next dose in the lower dose groups and
within 7-10 days after the hu1124 levels decreased to below
detection levels. In the two highest dose groups, hu1124 binding
sites remained saturated during the course of treatment.
[0083] A reversible increase in the average number of lymphocytes
was observed in the higher dose groups after Day 7 with a return to
the pretreatment numbers after dosing was completed. No effects on
the distribution of T and B subtypes or decreases in T-cell
subclasses were observed with any dose.
[0084] The tetanus antibody test results indicated that an
established humoral antibody response, especially a second set IgG
mediated antibody response, is able to persist in the presence of
multiple doses of hu1124. Antibody response was evaluated in 36 of
39 patients by double antigen sandwich ELISA. No human anti-hu1124
antibody response was detected out to Day 98 after multiple weekly
doses.
Results--Efficacy
[0085] Efficacy was based on the global assessment of improvement,
PASI scores, and histological analysis of skin biopsies. Some
clinical improvement (.gtoreq. poor improvement) was observed in
76% (29/38) of the subjects at Day 56. Of the subjects who received
at least 0.3 mg/kg/wk, 64% (18/28) experienced clinical improvement
of at least fair. Five subjects experienced an excellent
improvement (i.e., 75-90% improvement from baseline), with higher
rates of clinical improvement noted as the dose of study drug was
escalated. Continuous improvement at Day 70 was observed in six
subjects in the higher dose groups. Subjects infused with the
higher doses of hu1124 had greater decreases in their PASI scores
compared with subjects infused with the lower doses. Ten subjects
had a 50% decrease in their PASI scores. A dose response as
determined by decreasing PASI scores was noted in the higher dose
groups at Day 28; however, by Day 56 no substantial differences
were detected among the higher dose groups. Histological analysis
of the skin biopsies revealed significant reduction in epidermal
thickness and T-cell infiltration with clear anti-inflammatory
effects and reversal of pathological epidermal hyperplasia in
subjects treated with at least 0.3 mg/kg/wk.
Conclusions
[0086] The conclusions from this study of multiple, escalated
infusions of hu1124 in subjects with moderate to severe plaque
psoriasis are as follows: [0087] (a) hu1124 was safe and
well-tolerated in subjects who received multiple, escalated,
infused doses ranging from 0.1 mg/kg/every two weeks to 1.0
mg/kg/wk; [0088] (b) hu1124 provided clinical improvement in the
majority of subjects as measured by global assessments, PASI
scores, and histology; [0089] (c) hu1124 reduced the CD11a levels
on circulating lymphocytes; [0090] (d) hu1124 did not affect an
established humoral antibody response and did not elicit an immune
response; [0091] (e) hu1124 did not deplete lymphocyte counts;
lymphocyte counts increased during higher dose treatment.
Example 2
[0091] A Single-Dose and Multiple-Dose, Escalating Dose Study to
Evaluate the Safety Pharmacokinetics, and Biological Activity of
Subcutaneously Administered hu1124 in Subjects with Moderate to
Severe Plaque Psoriasis (HUPS254)
[0092] This study assessed the safety, pharmacokinetics and
pharmacodynamics, and biological activity of hu1124 administered by
subcutaneous injection in a single dose and in multiple doses to
subjects with moderate to severe plaque psoriasis.
[0093] The study drug, hu1124, was the same as used in Example 1.
It was supplied as single use vials containing 100 milligrams of
sterile, pyrogen-free, lyophilized drug product which contains
hu1124 antibody at a concentration of 100 mg/mL and 0.02 mmole
L-histidine, and 0.96 mmole, -trehalose, pH 6.0 when reconstituted
with 1.0 mL of sterile water for injection.
[0094] Twenty-six subjects received hu1124 subcutaneously. Subjects
in Group A (n=2) received a single injection of 0.3 mg/kg of
hu1124. Subjects in Groups B through E (n=24) received eight weekly
injections in doses ranging from 0.5-2.0 mg/kg of hu1124.
[0095] Table 2 below shows the treatment schedule and number of
patients in each arm of the study. TABLE-US-00002 TABLE 2 Dose
Escalation and Treatment Schedule Dose n (Actual Day Day Day Day
Day Day mg/kg Enrollment) Day 0 Day 7 14 21 28 35 42 49 Group A 2
0.3 -- -- -- -- -- -- -- (0.3) Group B 4 0.5 0.5 0.5 0.5 0.5 0.5
0.5 0.5 (0.5) Group C 6 0.5 0.7 1.0 1.0 1.0 1.0 1.0 1.0 (0.5-1.0)
Group D 6 0.7 1.0 1.5 1.5 1.5 1.5 1.5 1.5 (0.7-1.5) Group E 8 1.0
1.5 2.0 2.0 2.0 2.0 2.0 2.0 (1.0-2.0)
[0096] Adverse events were reported and assessed in a manner
similar to Example 1. Acute adverse events included headache,
fever, chills, myalgia, nausea, and vomiting. Any of these adverse
events occurring within 48 hours from the time of injection with
hu1124 were considered acute. Safety was assessed by pre- and
post-treatment examinations (including vital sign measurements),
clinical laboratory assessments (including blood chemistries,
hematology, and urinalysis), by examination of reported adverse
clinical events, and by a hearing assessment. Evaluation of
efficacy was based on PGA levels of improvements, changes in PASI
scores, and histological analysis of skin biopsies at Day 56.
Results--Safety
[0097] Subcutaneous administration of hu1124 has been very well
tolerated. No local cutaneous reaction has been observed. The
incidence of acute adverse reactions (occurring within 48 hours)
that were seen previously, after intravenous administration
(Example 1, HUPS249), appeared to have decreased by approximately
50%. The low incidence of headache (mild) in eight of 26 subjects
(31%) and fever (low grade) in two of 26 subjects (8%) was
noteworthy. Use of the "desensitizing" or "conditioning" dosing
schedule of the invention for dose administration has allowed the
safe administration of up to 2 mg/kg with minimal acute adverse
events.
Results--Efficacy
[0098] The study population was defined as subjects who had a
history of and/or were considered for systemic therapy for chronic
moderate to severe plaque-type psoriasis (BSA>15% and
PASI>12) which had been diagnosed for at least six months and
had been stable for at least three months.
[0099] On the basis of the Physicians' Global Assessment (PGA), and
decreases in Psoriasis Area Severity Index (PASI) score, a clear
benefit of hu1124 treatment was seen.
[0100] Improvement in psoriasis as judged by the PGA suggests a
dose response. In Group A, [single dose group (0.3 mg/kg)], no
improvement was seen, as expected. Subjects in Groups B, C, and D
showed some improvement at Day 56. The improvements were as
follows: in Group B (0.5 mg/kg), 1 Excellent (75-99% improvement
over baseline), 1 Slight (1-24% improvement), 2 Withdrawn; in Group
C (0.5-1.0 mg/kg), 2 Good (50-74% improvement), 1 Fair (25-49%
improvement), 3 Slight; in Group D (0.7-1.5 mg/kg), 1 Good, 1 Fair,
4 Slight. Subjects in Group E show the greatest amount of
improvement. In this group, were 1 Excellent, 2 Good, 2 Fair, 2
Slight, and 1 Withdrawn. The response in Group E indicates that all
subjects dosed with hu1124 showed some improvement (one subject
withdrew at Day 7).
[0101] Psoriasis Area and Severity Index (PASI) scores decreased by
an average of 10.9% in the single dose 0.3 mg/kg group, by 47.1% in
the 0.5 mg/kg group, by 36.3% in the 0.5-1.0 mg/kg group, by 33.2%
in the 0.7-1.5 mg/kg group and by 35.6%, in the 1.0-2.0 mg/kg
group. In general, the higher the PGA, the greater the reduction in
PASI score.
[0102] This study was later expanded to add 15 more subjects to
Group C and 16 more to Group E. Thus, Group C had a final
enrollment of 21 subjects and Group E had 24 subjects, adding to a
total of 55 subjects enrolled at 10 study centers (Group A study
was discontinued).
[0103] The data generated from the total pool of 55 subjects from
the multiple dose groups showed the following results and
observations. Substantial improvements in PGA and PASI scores were
observed by Day 56. Of the 55 subjects in the multiple-dose groups,
45% experienced Good or better improvements in PGA (defined as an
improvement of .gtoreq.50% of all clinical signs and symptoms of
psoriasis compared to baseline) and 47% experienced at least a 50%
decrease in PASI scores. Furthermore, 18% of these subjects
experienced at least a 75% reduction in PASI scores by Day 56 and
were categorized as treatment responders. Among the multiple dose
groups, higher proportions of subjects in the 0.5-1.0 mg/kg and
1.0-2.0 mg/kg groups experienced Good or better improvement in PGA.
Of note, one subject in the 1.0-2.0 mg/kg group experienced compete
resolution of disease symptoms as assessed by PGA. Higher
proportions of subjects in these two dose groups also experienced
at least 50% reductions in PASI scores compared with subjects in
the other dose groups.
Results--Summary
[0104] Administration of a subcutaneous formulation of hu1124 was
well-tolerated at the site of injection. Clinical benefits were
clearly seen with Fair, Good, and Excellent responses (PGA)
observed in several dosing groups. The most significant benefits
were seen in Group E (1.0-2.0 mg/kg), in particular, where all
subjects dosed (one subject withdrew) showed improvement as early
as Days 28-42 of therapy.
Example 3
An Extended Duration, Multiple-Dose Study to Evaluate the Safety,
Pharmacokinetics, and Biological Activity of Intravenously and
Subcutaneously Administered hu1124 in Subjects with Moderate to
Severe Plaque Psoriasis (HUPS256)
[0105] In this study, two treatment groups receive multiple
intravenous doses of hu1124 from 0.3 mg/kg to 1.0 mg/kg, for 12
weeks. Each hu1124 dose will be administered one time weekly over a
period of 90 minutes. Three treatment groups receive multiple
subcutaneous doses of hu1124 from 0.7 to 4.0 mg/kg, injected one
time weekly for 12 weeks.
[0106] For intravenous administration, the study drug, hu1124, is
supplied as a single-use, clear, colorless, sterile, non-pyrogenic
solution in a glass vial. Each vial contains 10 mL of solution at a
concentration of 4 mg/mL in 10 mM sodium acetate pH 5.0, with 0.02%
polysorbate 20, 0.1% sodium acetate trihydrate, and 4% mannitol. No
preservative is added to the solution. All study drug is stored at
2-8.degree. C. (35.6-46.4.degree. F.). For subcutaneous
administration, the study drug is supplied as single use vials
containing 100 milligrams of sterile, pyrogen-free, lyophilized
drug product which contains hu1124 antibody at a concentration of
100 mg/mL and 0.02 mmole L-histidine, and 0.96 mmole, -trehalose,
pH 6.0 when reconstituted with 1.0 mL of sterile water for
injection.
[0107] Table 3 below shows the treatment schedule and number of
patients in each arm of the study. TABLE-US-00003 TABLE 3 Treatment
Schedule hu1124 (mg/kg) DAY 0 7 14 21 28 35 42 49 56 63 70 77
Dosing Schedule (Subcutaneous Administration) Group A 0.3 0.3 0.3
0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 (n = 6) Group B 0.3 0.6 1.0 1.0
1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 (n = 10) Dosing Schedule
(Intravenous Administration) Group C 0.7 1.0 1.0 1.0 1.0 1.0 1.0
1.0 1.0 1.0 1.0 1.0 (n = 20) Group D 0.7 2.0 2.0 2.0 2.0 2.0 2.0
2.0 2.0 2.0 2.0 2.0 (n = 20) Group E 0.7 4.0 4.0 4.0 4.0 4.0 4.0
4.0 4.0 4.0 4.0 4.0 (n = 20)
[0108] Safety is assessed by pre- and post-treatment examinations
(including vital sign measurements), hearing assessments, clinical
laboratory assessments (including blood chemistries, hematology,
and urinalysis), antibody to hu1124 (HAHA), and by examination of
reported adverse clinical events as in the previous examples.
Pharmacokinetics are assessed using various ex vivo immunologic
assays. Biological activity is assessed by changes in PASI.
[0109] Results To date, 61 patients have been enrolled and the
dosing has been well tolerated at all doses used in the study. One
adverse event of local skin reaction has been reported among the 40
patients who have received SC doses up to 2.0 mg/kg. None of the 18
patients who have received SC doses up to 4.0 mg/kg have reported
an adverse event of local skin reaction. Clinical benefits are seen
with Fair, Good, and Excellent responses (PGA) observed. To date,
anti-CD11a (hu1124) administered by SC injection appears to be
safe, well tolerated, and exhibits promising biological and
clinical activity.
Example 4
Phase III Study of Efficacy, Safety, and Tolerability of
Subcutaneous Administration of hu1124 in Subjects with Moderate to
Severe Plaque Psoriasis during Three Phases: First Treatment,
Retreatment, and Extended Treatment (ACD2058g)
[0110] Subjects receive 12 weekly doses of anti-CD11a (hu1124) or
placebo administered by SC injection, as outlined in the Table of
dosing schedule immediately below. The doses consist of an initial
conditioning dose at a concentration of 0.7 mg/kg SC and weekly
doses of 1.0 mg/kg administered SC or 2.0 mg/kg SC thereafter.
TABLE-US-00004 TABLE 4 Dosing Schedule Day 0 7 14 21 28 35 42 49 56
63 70 77 Low dose 0.7 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0
(mg/kg) High dose 0.7 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0
(mg/kg)
[0111] Anti-CD11a (hu1124) is supplied as in the above Examples 2
and 3. When reconstituted with 1.0 mL of Sterile Water for
Injection (SWFI), each vial contains hu1124 at a concentration of
100 mg/mL, as well as polysorbate 20, L-histidine hydrochloride,
and .beta..beta.-trehalose, at pH of 6.0.
[0112] Primary efficacy determinations are made on FT Day 84, which
is the end of the First Treatment (FT) period. On FT Day 84,
subjects are defined as responders, partial responders, or
non-responders according to the following definitions: Responder:
PASI has decreased .gtoreq.75% from FT Day 0; Partial responder:
PASI has decreased .gtoreq.50% but <75% from FT Day 0;
Non-responder: PASI has decreased <50% from FT Day 0.
[0113] This response to therapy determines whether subjects are
assigned to the Observation (OB) or Extended Treatment (ET) period
after completion of the FT Day 84 assessments. Subjects defined as
responders enter the Observation (OB) period and are followed
either for 6 months or until relapse, whichever occurs first.
Relapse is defined as loss of 50% or more of the improvement in
PASI achieved between FT Day 0 and FT Day 84 (see Section 4.5.3.a).
At the time of relapse, subjects who received active drug during
the First Treatment (FT) period enter the Retreatment (RT) period
and are re-randomized in a 2:1 ratio to anti-CD11a or placebo,
respectively. Subjects who received placebo during the First
Treatment (FT) period and qualify as responders receive anti-CD11a
during the Retreatment period. Despite re-randomization, subjects
remain within the dose level group (low dose or high dose) assigned
during the First Treatment (FT) period. During the Retreatment (RT)
period, subjects receive a second course of treatment consisting of
12 weekly SC injections.
[0114] Subjects defined as partial responders or non-responders at
the end of the First Treatment (FT) period are assigned to the
Extended Treatment (ET) period. Subjects remain within the dose
levels assigned in the First Treatment (FT) period. Subjects who
had received anti-CD11a in the First Treatment (FT) period are
re-randomized 2:1 to anti-CD11a or placebo, respectively. All
subjects who received placebo in the First Treatment (FT) period
are assigned to anti-CD11a within their dose level. ET Day 0 occurs
on the same day as FT Day 84; hence the two courses of study drug
treatment are continuous over a 24-week period.
[0115] In a related study (ACD2062g), subjects who had received
prior anti-CD11a treatment and have antibodies to anti-CD11a, and
subjects who are receiving concurrent topical psoriasis therapy or
Ultraviolet B light (UVB) phototherapy are treated on the same
dosing regimen as shown in the preceding table.
[0116] Efficacy, safety and tolerability are measured as described
in the above examples.
Results
[0117] The above dosing regimen with initial, lower conditioning
dose is well tolerated. Clinical benefits with Fair, Good, and
Excellent responses (PGA) are observed.
Example 5
A Phase II Study to Evaluate the Safety and Efficacy of anti-CD11a
Antibody, hu1124 in Adults with Allergic Asthma Undergoing
Aeroallergen-Provoked Bronchoconstriction (ACD2017g)
[0118] This study evaluates the safety, tolerability, and efficacy
of eight weekly SC (subcutaneous) injections of up to 2.0 mg/kg
hu1124 in adult subjects with mild to moderate allergic asthma. The
effect of hu1124 on early and late asthmatic response to an inhaled
aeroallergen, and on both nonspecific and allergen-induced airway
hyperresponsiveness, are evaluated. An initial low, conditioning
dose of 0.7 mg/kg is administered followed by subsequent dosing at
2.0 mg/kg.
[0119] Thirty-six subjects with allergic asthma, ages 18-60 years,
are treated in this study. Cutaneous responses to allergen
(screened by skin prick testing and assessed by endpoint titration)
are measured, and methacholine bronchial challenges are conducted.
These data are used to estimate the starting allergen dose for use
in allergen bronchial challenge, using an empirically derived and
validated formula.
[0120] During the treatment period, study drug (hu1124 or placebo)
is administered weekly by SC injection, for eight doses over 50
days (one dose per week). The first dose of study drug is
administered on Day 0. Subjects undergo allergen bronchial and
methacholine challenges. Methacholine challenges, measuring
nonspecific airway hyperresponsiveness (basal and post-allergen),
are performed the day prior to a scheduled allergen challenge and
.about.24 hours following the start of allergen challenge.
[0121] The hu1124 antibody is supplied by Genentech as a sterile,
pyrogen-free, lyophilized drug product in 10-mL glass vials. When
reconstituted with 1.0 mL of Sterile Water for Injection (SWFI),
each vial contains hu1124 at a concentration of 100 mg/mL, 20 mmol
of L-histidine, 240 mmol of .beta.,.beta.-trehalose, 0.04%
polysorbate 20 (pH of 6.0). The placebo formulation has the same
product composition but does not contain hu1124. Study drug is
administered by SC injection in the forearm, thigh, or abdomen. The
dosing regimen is shown in the table immediately below.
TABLE-US-00005 TABLE 5 Dosing Regimen Treat- Day ment 0 7 14 21 28
35 42 49 hu1124 0.7 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Placebo x x x x x x
x x Doses of hu1124 are given in mg/kg.
[0122] The safety and efficacy of hu1124 are assessed by the
incidence and severity of adverse events, laboratory tests,
physical examinations including vital signs, spirometry, and serum
antibody response to hu1124 at baseline and during the treatment
and follow-up periods. The incidence and magnitude of changes in
hearing are assessed by audiograms. Efficacy measures include
determining the change in LAR and EAR and allergen-induced increase
in airway responsiveness. All subjects are followed for 28 days
after dosing is complete.
[0123] Blood and urine samples are collected periodically for
analysis of blood chemistries, hematology, and urinalysis. Specific
parameters assessed are as follows: Chemistry: sodium, potassium,
chloride, bicarbonate, glucose, BUN, creatinine, calcium,
phosphorus, magnesium, total and direct bilirubin, albumin, ALT,
AST, alkaline phosphatase, uric acid, total protein. Hematology:
CBC with differential and platelet count. Urinalysis: complete
urinalysis with microscopic examination. Serum antibodies to
hu1124. Serum samples for pharmacokinetic evaluations. Vital sign
measurement consists of sitting blood pressure, respiratory rate,
pulse rate, and body temperature measured orally (.degree. C.).
[0124] The above dosing regimen is well tolerated and shows
efficacy in treating asthma.
Example 6
[0125] Phase I/II Study of Anti-CD11a (hu1124) for Renal
Transplantation
[0126] The use of a non-T cell-depleting, humanized, monoclonal
anti-CD11a antibody should cause significantly less toxicity,
sensitization, and more specific immunosuppression than currently
available anti-T cell monoclonal antibodies (OKT3).
[0127] This study evaluates the safety, pharmacokinetics and
pharmacodynamics (PK/PD), biological activity and clinical effects
of 12 weeks of subcutaneously administered anti-CD11a (hu1124) at 2
dose levels in patients undergoing their first cadaveric donor
kidney transplant. Patients are treated with 12 doses at weekly
intervals and followed for 13 weeks. Each dose group has two arms
in which standard background immunosuppression regimens are used,
as summarized in Study Design Table 5 below. The immunosuppressive
agents are cyclosporine, prednisone, mycophenolate mofetil (MMF)
and sirolimus. Each patient receives the initial "conditioning"
dose of anti-CD11a (hu1124) at 0.5 mg/kg or 0.7 mg/kg, dependent on
dose group assignment, on Day 0 at least 1 hour prior to surgery as
summarized in Table 6 below. Thereafter, each patient receives a
maintenance dose of anti-CD11a (hu1124) at 0.5 mg/kg or 2.0 mg/kg
each weekly visit for 11 weeks, based on the dose group.
TABLE-US-00006 TABLE 6 Study Design Dose Group n Group I Anti-CD11a
(hu1124) 0.5 mg/kg plus Arm A. Half-dose cyclosporine (2.5-5
mg/kg), sirolimus, 9 prednisone Arm B. Full-dose cyclosporine
(>5-10 mg/kg), MMF, 9 prednisone Group II Anti-CD11a (hu1124)
2.0 mg/kg plus Arm A. Half-dose cyclosporine (2.5-5 mg/kg),
sirolimus, 9 prednisone Arm B. Full-dose cyclosporine (>5-10
mg/kg), MMF, 9 prednisone
[0128] TABLE-US-00007 TABLE 7 Dosing Regimen Days of Study Dose
Group 0 7 14 21 28 35 42 49 56 63 70 77 Group I: 0.5 0.5 0.5 0.5
0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 mg/kg Group II: 2.0 0.7 2.0 2.0
2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 mg/kg
[0129] The study drug, anti-CD11a (hu1124), is supplied by XOMA as
a sterile, pyrogen-free, lyophilized drug product in 10 mL glass
vials which, when reconstituted with 1.0 mL Sterile Water for
Injection (SWFI), contain anti-CD11a (hu1124) at a concentration of
100 mg/mL, 20 mmol of L-histidine, 240 mmol of
.alpha.,.alpha.-trehalose, and 0.04% polysorbate 20 (pH of 6.0).
Vials are refrigerated at 2.degree. C.-8.degree. C. (36.degree.
F.-46.degree. F.). Reconstituted product is stable at room
temperature for up to 8 hours.
[0130] Safety is assessed by the incidence of adverse events, vital
signs, changes in laboratory values compared with baseline
(hematology, chemistry, urinalysis), infections, lymphoma, acute
rejection episodes, delayed graft function, graft loss, death,
incidence of human anti-humanized monoclonal antibody (HAHA).
[0131] After initial dosing of the study drug in each patient, the
pharmacokinetic characteristics of the study drug are assessed by
serial measurement of plasma concentrations of anti-CD11a (hu1124)
at various time points throughout the duration of the study. Flow
cytometry analysis of T cells, T lymphocyte subsets (CD3/CD4/CD8),
NK cells, and B lymphocytes, plus expression of CD11a on T
lymphocytes, are performed. Gene activation in cytotoxic T
lymphocytes (perforin, granzyme B, FAS L) and cytokine gene
expression (IL-10, IFN gamma, IL-2, TGF beta, IL-13) in urinary
lymphocytes (Vasconcellos et al. 1998, Transplantation 66: 562-566)
are examined. In addition, the humoral immune response is studied
by examining peripheral blood for anti-HLA antibody production, by
ELISA.
[0132] The biological activity is measured as follows. Alterations
in graft function are assessed by serial measurements of serum
creatinine, the need for dialysis during the initial 7 days post
transplant, development of proteinuria, acute rejection episodes,
and response to high-dose steroid therapy (as measured by serum
creatinine). As per transplant center standard of care, graft
biopsies are obtained at the time of implantation, when necessary
to confirm the diagnosis of acute rejection, or if rejection does
not occur, 1 week after treatment completion (Day 84). Material is
obtained for cytokine and cytotoxic T lymphocyte mRNA analysis,
quantitation of interstitial fibrosis, and other potential markers
of chronic allograft nephropathy (i.e., collagen expression).
[0133] The above dosing regimen with an initial, lower conditioning
dose of anti-CD11a antibody, followed by a higher therapeutic dose
reduces the incidence of both acute graft rejection and delayed
graft function, and promotes long-term survival with minimum
toxicity, compared to the administration of equal doses
throughout.
[0134] The method of the invention provides a higher therapeutic
index than conventional and current therapy by minimizing toxicity
and adverse side effects. The foregoing written specification is
considered to be sufficient to enable one skilled in the art to
practice the invention. Indeed, various modifications of the
invention in addition to those shown and described herein will
become apparent to those skilled in the art from the foregoing
description and fall within the scope of the appended claims.
* * * * *