U.S. patent application number 17/174019 was filed with the patent office on 2022-04-07 for methods of treating cancer using pd-1 axis binding antagonists and mek inhibitors.
This patent application is currently assigned to Genentech, Inc.. The applicant listed for this patent is Genentech, Inc.. Invention is credited to Melissa JUNTTILA.
Application Number | 20220105180 17/174019 |
Document ID | / |
Family ID | |
Filed Date | 2022-04-07 |
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United States Patent
Application |
20220105180 |
Kind Code |
A1 |
JUNTTILA; Melissa |
April 7, 2022 |
METHODS OF TREATING CANCER USING PD-1 AXIS BINDING ANTAGONISTS AND
MEK INHIBITORS
Abstract
The present invention describes combination treatment comprising
a PD-1 axis binding antagonist and a MEK inhibitor and methods for
use thereof, including methods of treating conditions where
enhanced immunogenicity is desired such as increasing tumor
immunogenicity for the treatment of cancer.
Inventors: |
JUNTTILA; Melissa; (South
San Francisco, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Genentech, Inc. |
South San Francisco |
CA |
US |
|
|
Assignee: |
Genentech, Inc.
South San Francisco
CA
|
Appl. No.: |
17/174019 |
Filed: |
February 11, 2021 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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15399118 |
Jan 5, 2017 |
10946093 |
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17174019 |
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PCT/US2015/040582 |
Jul 15, 2015 |
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15399118 |
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62024988 |
Jul 15, 2014 |
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International
Class: |
A61K 39/395 20060101
A61K039/395; C07K 16/28 20060101 C07K016/28; A61K 31/4523 20060101
A61K031/4523 |
Claims
1-53. (canceled)
54. A method for treating or delaying progression of cancer in an
individual comprising administering to the individual an effective
amount of a PD-1 axis binding antagonist and a MEK inhibitor,
wherein the individual has not been previously treated with a B-raf
antagonist, and wherein the individual has cancer that is at risk
of developing resistance to the B-raf antagonist.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the priority benefit of U.S.
provisional application Ser. No. 62/024,988, filed Jul. 15, 2014,
the contents of which are incorporated herein by reference in its
entirety.
SUBMISSION OF SEQUENCE LISTING ON ASCII TEXT FILE
[0002] The content of the following submission on ASCII text file
is incorporated herein by reference in its entirety: a computer
readable form (CRF) of the Sequence Listing (file name:
146392027540SeqList.txt, date recorded: Jul. 8, 2015, size: 22
KB).
BACKGROUND
[0003] The provision of two distinct signals to T-cells is a widely
accepted model for lymphocyte activation of resting T lymphocytes
by antigen-presenting cells (APCs). Lafferty et al, Aust. J. Exp.
Biol. Med. ScL 53: 27-42 (1975). This model further provides for
the discrimination of self from non-self and immune tolerance.
Bretscher et al, Science 169: 1042-1049 (1970); Bretscher, P. A.,
P.N.A.S. USA 96: 185-190 (1999); Jenkins et al, J. Exp. Med. 165:
302-319 (1987). The primary signal, or antigen specific signal, is
transduced through the T-cell receptor (TCR) following recognition
of foreign antigen peptide presented in the context of the major
histocompatibility-complex (MHC). The second or co-stimulatory
signal is delivered to T-cells by co-stimulatory molecules
expressed on antigen-presenting cells (APCs), and induces T-cells
to promote clonal expansion, cytokine secretion and effector
function. Lenschow et al., Ann. Rev. Immunol. 14:233 (1996). In the
absence of co-stimulation, T-cells can become refractory to antigen
stimulation, do not mount an effective immune response, and further
may result in exhaustion or tolerance to foreign antigens.
[0004] In the two-signal model T-cells receive both positive and
negative secondary co-stimulatory signals. The regulation of such
positive and negative signals is critical to maximize the host's
protective immune responses, while maintaining immune tolerance and
preventing autoimmunity. Negative secondary signals seem necessary
for induction of T-cell tolerance, while positive signals promote
T-cell activation. While the simple two-signal model still provides
a valid explanation for naive lymphocytes, a host's immune response
is a dynamic process, and co-stimulatory signals can also be
provided to antigen-exposed T-cells. The mechanism of
co-stimulation is of therapeutic interest because the manipulation
of co-stimulatory signals has shown to provide a means to either
enhance or terminate cell-based immune response. Recently, it has
been discovered that T cell dysfunction or anergy occurs
concurrently with an induced and sustained expression of the
inhibitory receptor, programmed death 1 polypeptide (PD-1). As a
result, therapeutic targeting of PD-1 and other molecules which
signal through interactions with PD-1, such as programmed death
ligand 1 (PD-L1) and programmed death ligand 2 (PD-L2) are an area
of intense interest.
[0005] PD-L1 is overexpressed in many cancers and is often
associated with poor prognosis (Okazaki T et al., Intern. Immun.
2007 19(7):813) (Thompson R H et al., Cancer Res 2006, 66(7):3381).
Interestingly, the majority of tumor infiltrating T lymphocytes
predominantly express PD-1, in contrast to T lymphocytes in normal
tissues and peripheral blood T lymphocytes indicating that
up-regulation of PD-1 on tumor-reactive T cells can contribute to
impaired antitumor immune responses (Blood 2009 114(8): 1537). This
may be due to exploitation of PD-L1 signaling mediated by PD-L1
expressing tumor cells interacting with PD-1 expressing T cells to
result in attenuation of T cell activation and evasion of immune
surveillance (Sharpe et al., Nat Rev 2002) (Keir M E et al., 2008
Annu. Rev. Immunol. 26:677). Therefore, inhibition of the
PD-L1/PD-1 interaction may enhance CD8+ T cell-mediated killing of
tumors.
[0006] The inhibition of PD-1 axis signaling through its direct
ligands (e.g., PD-L1, PD-L2) has been proposed as a means to
enhance T cell immunity for the treatment of cancer (e.g., tumor
immunity). Moreover, similar enhancements to T cell immunity have
been observed by inhibiting the binding of PD-L1 to the binding
partner B7-1. Furthermore, combining inhibition of PD-1 signaling
with other signaling pathways (e.g. MAPK pathway, "MEK") that are
deregulated in tumor cells may further enhance treatment efficacy.
However, an optimal therapeutic treatment would combine blockade of
PD-1 receptor/ligand interaction with an agent that directly
inhibited tumor growth, optionally further including unique immune
enhancing properties not provided by PD-1 blockade alone. There
remains a need for such an optimal therapy for treating,
stabilizing, preventing, and/or delaying development of various
cancers.
[0007] All references, publications, and patent applications
disclosed herein are hereby incorporated by reference in their
entirety.
BRIEF SUMMARY
[0008] Provided herein are methods for treating or delaying
progression of cancer in an individual comprising administering to
the individual an effective amount of a PD-1 axis binding
antagonist and a MEK inhibitor, wherein the individual has cancer
or is at risk of developing cancer that is resistant to a B-raf
antagonist. In some embodiments, the method further comprises
diagnosing the individual as having a cancer that is resistant to a
B-raf antagonist, wherein the diagnosing occurs prior to
administering the effective amount of the PD-1 axis binding
antagonist and the MEK inhibitor. In some embodiments, the method
further comprises selecting an individual for treatment based on
the individual having cancer that is resistant to a B-raf
antagonist or assessing that the individual is at risk of
developing cancer that is resistant to a B-raf antagonist, wherein
the selecting occurs prior to administering the effective amount of
the PD-1 axis binding antagonist and the MEK inhibitor. In some
embodiments, the individual has not been previously treated with a
B-raf antagonist. In some embodiments, the individual has been
previously treated with a B-raf antagonist.
[0009] In another aspect, provided herein are methods for treating
or delaying progression of cancer in an individual comprising (a)
diagnoising the individual as having a cancer that is resistant to
a B-raf antagonist; and (b) administering to the individual an
effective amount of a PD-1 axis binding antagonist and a MEK
inhibitor, wherein the administering occurs after diagnosing the
individual. In some embodiments, the individual has not been
previously treated with a B-raf antagonist. In some embodiments,
the individual has been previously treated with a B-raf
antagonist.
[0010] In another aspect, provided herein are methods for treating
or delaying progression of cancer in an individual comprising (a)
selecting an individual for treatment based on the individual
having cancer that is resistant to a B-raf antagonist or assessing
that the individual is at risk of developing cancer that is
resistant to a B-raf antagonist; and (b) administering to the
individual an effective amount of a PD-1 axis binding antagonist
and a MEK inhibitor, wherein the administering occurs after
selecting the individual. In some embodiments, the individual has
not been previously treated with a B-raf antagonist. In some
embodiments, the individual has been previously treated with a
B-raf antagonist.
[0011] In another aspect, provided herein are methods for treating
or delaying progression of cancer in an individual comprising
administering to the individual an effective amount of a PD-1 axis
binding antagonist and a MEK inhibitor, wherein the individual has
been previously treated with a B-raf antagonist for cancer.
[0012] In some embodiments, the cancer in the individual has
progressed within 1 month, 6 months, 1 year, or 5 years after
completing a B-raf antagonist-based therapy regimen. In some
embodiments, the B-raf antagonist is a small molecule inhibitor, an
antibody, a peptide, a peptidomimetic, an aptamer or a
polynucleotide. In some embodiments, the B-raf antagonist is
dabrafenib, vemurafenib, GSK 2118436, RAF265, XL281, ARQ736,
BAY73-4506, sorafenib, PLX4720, PLX-3603, GSK2118436, GDC-0879, or
N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-dif-
luorophenyl)propane-1-sulfonamide. In some embodiments, the B-raf
antagonist is a selective B-raf antagonist of B-raf V600. In some
embodiments, the selective B-raf antagonist of B-raf V600 is a
selective antagonist of B-raf V600E. In some embodiments, the
selective B-raf antagonist of B-raf V600 is a selective antagonist
of B-raf V600E, B-raf V600K, and/or V600D. In some embodiments, the
selective B-raf antagonist of B-raf V600 is a selective antagonist
of B-raf V600R.
[0013] In some embodiments, the cancer contains a BRAF V600E
mutation, a BRAF wildtype, a KRAS wildtype, or an activating KRAS
mutation. In some embodiments, the treatment results in a sustained
response in the individual after cessation of the treatment. In
some embodiments, the individual has colorectal cancer, melanoma,
lung cancer, ovarian cancer, breast cancer, pancreatic cancer,
hematological malignancy, bladder cancer, and/or renal cell
carcinoma. In some embodiments, the cancer is metastatic.
[0014] In some embodiments, the PD-1 axis binding antagonist is
selected from the group consisting of a PD-1 binding antagonist, a
PD-F1 binding antagonist and a PD-F2 binding antagonist. In some
embodiments, the PD-1 axis binding antagonist is a PD-1 binding
antagonist. In some embodiments, the PD-1 binding antagonist
inhibits the binding of PD-1 to its ligand binding partners. In
some embodiments, the PD-1 binding antagonist inhibits the binding
of PD-1 to PD-F1, PD-1 to PD-F2, or PD-1 to both PD-F1 and PD-F2.
In some embodiments, the PD-1 binding antagonist is an antibody. In
some embodiments, the PD-1 binding antagonist is MDX-1106, Merck
3745, CT-011, MEDI-0680, PDR001, REGN2810, BGB-108, BGB-A317, or
AMP-224. In some embodiments, the PD-1 binding antagonist is
nivolumab, pembrolizumab, pidilizumab, MEDI-0680, PDR001, REGN2810,
BGB-108, BGB-A317, or AMP-224. In some embodiments, the PD-1 axis
binding antagonist is a PD-L1 binding antagonist. In some
embodiments, the PD-L1 binding antagonist inhibits the binding of
PD-L1 to PD-1, PD-L1 to B7-1, or PD-L1 to both PD-1 and B7-1. In
some embodiments, the PD-L1 binding antagonist is an anti-PD-L1
antibody. In some embodiments, the anti-PD-L1 antibody is a
monoclonal antibody. In some embodiments, the anti-PD-L1 antibody
is an antibody fragment selected from the group consisting of Fab,
Fab'-SH, Fv, scFv, and (Fab').sub.2 fragments. In some embodiments,
the anti-PD-L1 antibody is a humanized antibody or a human
antibody. In some embodiments, the PD-L1 binding antagonist is
selected from the group consisting of: YW243.55.S70, MPDL3280A,
MEDI4736, MDX-1105, and MSB0010718C. In some embodiments, the PD-L1
binding antagonist is selected from the group consisting of:
YW243.55.S70, atezolizumab, durvalumab, MDX-1105, and avelumab. In
some embodiments, the antibody comprises a heavy chain comprising
HVR-H1 sequence of SEQ ID NO: 15, HVR-H2 sequence of SEQ ID NO:16,
and HVR-H3 sequence of SEQ ID NO:3; and a light chain comprising
HVR-L1 sequence of SEQ ID NO: 17, HVR-L2 sequence of SEQ ID NO: 18,
and HVR-L3 sequence of SEQ ID NO: 19. In some embodiments, the
antibody comprises a heavy chain variable region comprising the
amino acid sequence of SEQ ID NO:24 or 28 and a light chain
variable region comprising the amino acid sequence of SEQ ID NO:21.
In some embodiments, the PD-1 axis binding antagonist is a PD-L2
binding antagonist. In some embodiments, the PD-L2 binding
antagonist is an antibody. In some embodiments, the PD-L2 binding
antagonist is an immunoadhesin. In some embodiments, the MEK
inhibitor is a competitive inhibitor of MEK. In some embodiments,
the MEK inhibitor is more selective against an activating KRAS
mutation. In some embodiments, the MEK inhibitor is an allosteric
inhibitor of MEK. In some embodiments, the MEK inhibitor is more
selective against an activating BRAF mutation. In some embodiments,
the MEK inhibitor is a compound of the formula (I), (II), (III),
(IV), (V), (VI) or (VII), or a pharmaceutically acceptable salt or
solvate thereof. In some embodiments, the MEK inhibitor is selected
from the group consisting of G02442104, G-38963, G02443714,
G00039805 and GDC-0973, or a pharmaceutically acceptable salt or
solvate thereof. In some embodiments, the MEK inhibitor is
G02443714, G02442104 or G00039805.
[0015] In some embodiments, the MEK inhibitor is administered
continuously. In some embodiments, the MEK inhibitor is
administered intermittently. In some embodiments, the MEK inhibitor
is administered before the PD-1 axis binding antagonist. In some
embodiments, the MEK inhibitor is administered simultaneous with
the PD-1 axis binding antagonist. In some embodiments, the MEK
inhibitor is administered after the PD-1 axis binding antagonist.
In some embodiments, the PD-1 axis binding antagonist and/or the
MEK inhibitor is administered intravenously, intramuscularly,
subcutaneously, topically, orally, transdermally,
intraperitoneally, intraorbitally, by implantation, by inhalation,
intrathecally, intraventricularly, or intranasally.
[0016] In another aspect, provided herein are kits comprising a
PD-1 axis binding antagonist and a package insert comprising
instructions for using the PD-1 axis binding antagonist in
combination with a MEK inhibitor to treat or delay progression of
cancer in an individual, wherein the individual has cancer or is at
risk of developing cancer that is resistant to a B-raf antagonist.
In another aspect, provided herein are kits comprising a PD-1 axis
binding antagonist and a MEK inhibitor, and a package insert
comprising instructions for using the PD-1 axis binding antagonist
and the MEK inhibitor to treat or delay progression of cancer in an
individual, wherein the individual has cancer or is at risk of
developing cancer that is resistant to a B-raf antagonist. In
another aspect, provided herein are kits comprising a MEK inhibitor
and a package insert comprising instructions for using the MEK
inhibitor in combination with a PD-1 axis binding antagonist to
treat or delay progression of cancer in an individual, wherein the
individual has cancer or is at risk of developing cancer that is
resistant to a B-raf antagonist. In another aspect, provided herein
are kits comprising a PD-1 axis binding antagonist and a package
insert comprising instructions for using the PD-1 axis binding
antagonist in combination with a MEK inhibitor to treat or delay
progression of cancer in an individual, wherein the individual has
been previously treated with a B-raf antagonist for cancer. In
another aspect, provided herein are kits comprising a PD-1 axis
binding antagonist and a MEK inhibitor, and a package insert
comprising instructions for using the PD-1 axis binding antagonist
and the MEK inhibitor to treat or delay progression of cancer in an
individual, wherein the individual has been previously treated with
a B-raf antagonist for cancer. In another aspect, provided herein
are kits comprising a MEK inhibitor and a package insert comprising
instructions for using the MEK inhibitor in combination with a PD-1
axis binding antagonist to treat or delay progression of cancer in
an individual, wherein the individual has been previously treated
with a B-raf antagonist for cancer.
[0017] In some embodiments, the individual is a human.
[0018] It is to be understood that one, some, or all of the
properties of the various embodiments described herein may be
combined to form other embodiments of the present invention. These
and other aspects of the invention will become apparent to one of
skill in the art. These and other embodiments of the invention are
further described by the detailed description that follows.
BRIEF DESCRIPTION OF THE DRAWINGS
[0019] FIG. 1 shows tumor re-growth upon treatment with anti-PDL1,
a MEK inhibitor, or both. The graph shows the percent tumor volume
change over time during treatment with the indicated agent(s).
[0020] FIG. 2 shows individual animal responses to treatment with
Vemurafenib followed by anti-PDL1, a MEK inhibitor, or both. Each
bar depicts the percent change in tumor growth upon crossover from
Vemurafenib to the indicated treatment in an individual animal.
DETAILED DESCRIPTION
I. General Techniques
[0021] The techniques and procedures described or referenced herein
are generally well understood and commonly employed using
conventional methodology by those skilled in the art, such as, for
example, the widely utilized methodologies described in Sambrook et
al., Molecular Cloning: A Laboratory Manual 3d edition (2001) Cold
Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Current
Protocols in Molecular Biology (F. M. Ausubel, et al. eds.,
(2003)); the series Methods in Enzymology (Academic Press, Inc.):
PCR 2: A Practical Approach (MJ. MacPherson, B. D. Hames and G. R.
Taylor eds. (1995)), Harlow and Lane, eds. (1988) Antibodies, A
Laboratory Manual, and Animal Cell Culture (R. I. Freshney, ed.
(1987)); Oligonucleotide Synthesis (MJ. Gait, ed., 1984); Methods
in Molecular Biology, Humana Press; Cell Biology: A Laboratory
Notebook (J. E. Cellis, ed., 1998) Academic Press; Animal Cell
Culture (R. I. Freshney), ed., 1987); Introduction to Cell and
Tissue Culture (J. P. Mather and P. E. Roberts, 1998) Plenum Press;
Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J. B.
Griffiths, and D. G. Newell, eds., 1993-8) J. Wiley and Sons;
Handbook of Experimental Immunology (D. M. Weir and C. C.
Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J. M.
Miller and M. P. Calos, eds., 1987); PCR: The Polymerase Chain
Reaction, (Mullis et al., eds., 1994); Current Protocols in
Immunology (J. E. Coligan et al., eds., 1991); Short Protocols in
Molecular Biology (Wiley and Sons, 1999); Immunobiology (C. A.
Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997);
Antibodies: A Practical Approach (D. Catty., ed., IRL Press,
1988-1989); Monoclonal Antibodies: A Practical Approach (P.
Shepherd and C. Dean, eds., Oxford University Press, 2000); Using
Antibodies: A Laboratory Manual (E. Harlow and D. Lane (Cold Spring
Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.
D. Capra, eds., Harwood Academic Publishers, 1995); and Cancer:
Principles and Practice of Oncology (V. T. DeVita et al., eds.,
J.B. Lippincott Company, 1993).
II. Definitions
[0022] The term "antagonist" is used in the broadest sense, and
includes any molecule that partially or fully blocks, inhibits, or
neutralizes a biological activity of a native polypeptide disclosed
herein. In a similar manner, the term "agonist" is used in the
broadest sense and includes any molecule that mimics a biological
activity of a native polypeptide disclosed herein. Suitable agonist
or antagonist molecules specifically include agonist or antagonist
antibodies or antibody fragments, fragments or amino acid sequence
variants of native polypeptides, peptides, antisense
oligonucleotides, small organic molecules, etc. Methods for
identifying agonists or antagonists of a polypeptide may comprise
contacting a polypeptide with a candidate agonist or antagonist
molecule and measuring a detectable change in one or more
biological activities normally associated with the polypeptide.
[0023] The term "aptamer" refers to a nucleic acid molecule that is
capable of binding to a target molecule, such as a polypeptide. For
example, an aptamer of the invention can specifically bind to a
B-raf polypeptide, or to a molecule in a signaling pathway that
modulates the expression or activity of B-raf. The generation and
therapeutic use of aptamers are well established in the art. See,
e.g., U.S. Pat. No. 5,475,096, and the therapeutic efficacy of
Macugen.RTM. (Eyetech, New York) for treating age-related macular
degeneration.
[0024] The term "PD-1 axis binding antagonist" is a molecule that
inhibits the interaction of a PD-1 axis binding partner with either
one or more of its binding partner, so as to remove T-cell
dysfunction resulting from signaling on the PD-1 signaling
axis--with a result being to restore or enhance T-cell function
(e.g., proliferation, cytokine production, target cell killing). As
used herein, a PD-1 axis binding antagonist includes a PD-1 binding
antagonist, a PD-L1 binding antagonist and a PD-L2 binding
antagonist.
[0025] The term "PD-1 binding antagonists" is a molecule that
decreases, blocks, inhibits, abrogates or interferes with signal
transduction resulting from the interaction of PD-1 with one or
more of its binding partners, such as PD-L1, PD-L2. In some
embodiments, the PD-1 binding antagonist is a molecule that
inhibits the binding of PD-1 to its binding partners. In a specific
aspect, the PD-1 binding antagonist inhibits the binding of PD-1 to
PD-L1 and/or PD-L2. For example, PD-1 binding antagonists include
anti-PD-1 antibodies, antigen binding fragments thereof,
immunoadhesins, fusion proteins, oligopeptides and other molecules
that decrease, block, inhibit, abrogate or interfere with signal
transduction resulting from the interaction of PD-1 with PD-L1
and/or PD-L2. In one embodiment, a PD-1 binding antagonist reduces
the negative co-stimulatory signal mediated by or through cell
surface proteins expressed on T lymphocytes mediated signaling
through PD-1 so as render a dysfunctional T-cell less dysfunctional
(e.g., enhancing effector responses to antigen recognition). In
some embodiments, the PD-1 binding antagonist is an anti-PD-1
antibody. In a specific aspect, a PD-1 binding antagonist is
MDX-1106 described herein. In another specific aspect, a PD-1
binding antagonist is Merck 3745 described herein. In another
specific aspect, a PD-1 binding antagonist is CT-011 described
herein.
[0026] The term "PD-L1 binding antagonists" is a molecule that
decreases, blocks, inhibits, abrogates or interferes with signal
transduction resulting from the interaction of PD-L1 with either
one or more of its binding partners, such as PD-1, B7-1. In some
embodiments, a PD-L1 binding antagonist is a molecule that inhibits
the binding of PD-L1 to its binding partners. In a specific aspect,
the PD-L1 binding antagonist inhibits binding of PD-L1 to PD-1
and/or B7-1. In some embodiments, the PD-L1 binding antagonists
include anti-PD-L1 antibodies, antigen binding fragments thereof,
immunoadhesins, fusion proteins, oligopeptides and other molecules
that decrease, block, inhibit, abrogate or interfere with signal
transduction resulting from the interaction of PD-L1 with one or
more of its binding partners, such as PD-1, B7-1. In one
embodiment, a PD-L1 binding antagonist reduces the negative
co-stimulatory signal mediated by or through cell surface proteins
expressed on T lymphocytes mediated signaling through PD-L1 so as
to render a dysfunctional T-cell less dysfunctional (e.g.,
enhancing effector responses to antigen recognition). In some
embodiments, a PD-L1 binding antagonist is an anti-PD-L1 antibody.
In a specific aspect, an anti-PD-L1 antibody is YW243.55.S70
described herein. In another specific aspect, an anti-PD-L1
antibody is MDX-1105 described herein. In still another specific
aspect, an anti-PD-L1 antibody is MPDL3280A (atezolizumab)
described herein.
[0027] The term "PD-E2 binding antagonists" is a molecule that
decreases, blocks, inhibits, abrogates or interferes with signal
transduction resulting from the interaction of PD-L2 with either
one or more of its binding partners, such as PD-1. In some
embodiments, a PD-L2 binding antagonist is a molecule that inhibits
the binding of PD-L2 to its binding partners. In a specific aspect,
the PD-L2 binding antagonist inhibits binding of PD-L2 to PD-1. In
some embodiments, the PD-L2 antagonists include anti-PD-L2
antibodies, antigen binding fragments thereof, immunoadhesins,
fusion proteins, oligopeptides and other molecules that decrease,
block, inhibit, abrogate or interfere with signal transduction
resulting from the interaction of PD-L2 with either one or more of
its binding partners, such as PD-1. In one embodiment, a PD-L2
binding antagonist reduces the negative co-stimulatory signal
mediated by or through cell surface proteins expressed on T
lymphocytes mediated signaling through PD-L2 so as render a
dysfunctional T-cell less dysfunctional (e.g., enhancing effector
responses to antigen recognition). In some embodiments, a PD-L2
binding antagonist is an immunoadhesin.
[0028] The term "dysfunction" in the context of immune dysfunction,
refers to a state of reduced immune responsiveness to antigenic
stimulation. The term includes the common elements of both
exhaustion and/or anergy in which antigen recognition may occur,
but the ensuing immune response is ineffective to control infection
or tumor growth.
[0029] The term "dysfunctional", as used herein, also includes
refractory or unresponsive to antigen recognition, specifically,
impaired capacity to translate antigen recognition into down-stream
T-cell effector functions, such as proliferation, cytokine
production (e.g., IL-2) and/or target cell killing.
[0030] The term "anergy" refers to the state of unresponsiveness to
antigen stimulation resulting from incomplete or insufficient
signals delivered through the T-cell receptor (e.g. increase in
intracellular Ca.sup.+2 in the absence of ras-activation). T cell
anergy can also result upon stimulation with antigen in the absence
of co-stimulation, resulting in the cell becoming refractory to
subsequent activation by the antigen even in the context of
costimulation. The unresponsive state can often be overriden by the
presence of Interleukin-2. Anergic T-cells do not undergo clonal
expansion and/or acquire effector functions.
[0031] The term "exhaustion" refers to T cell exhaustion as a state
of T cell dysfunction that arises from sustained TCR signaling that
occurs during many chronic infections and cancer. It is
distinguished from anergy in that it arises not through incomplete
or deficient signaling, but from sustained signaling. It is defined
by poor effector function, sustained expression of inhibitory
receptors and a transcriptional state distinct from that of
functional effector or memory T cells. Exhaustion prevents optimal
control of infection and tumors. Exhaustion can result from both
extrinsic negative regulatory pathways (e.g., immunoregulatory
cytokines) as well as cell intrinsic negative regulatory
(costimulatory) pathways (PD-1, B7-H3, B7-H4, etc.).
[0032] "Enhancing T-cell function" means to induce, cause or
stimulate a T-cell to have a sustained or amplified biological
function, or renew or reactivate exhausted or inactive T-cells.
Examples of enhancing T-cell function include: increased secretion
of .gamma.-interferon from CD8.sup.+ T-cells, increased
proliferation, increased antigen responsiveness (e.g., viral,
pathogen, or tumor clearance) relative to such levels before the
intervention. In one embodiment, the level of enhancement is as
least 50%, alternatively 60%, 70%, 80%, 90%, 100%, 120%, 150%, or
200%. The manner of measuring this enhancement is known to one of
ordinary skill in the art.
[0033] A "T cell dysfunctional disorder" is a disorder or condition
of T-cells characterized by decreased responsiveness to antigenic
stimulation. In a particular embodiment, a T-cell dysfunctional
disorder is a disorder that is specifically associated with
inappropriate increased signaling through PD-1. In another
embodiment, a T-cell dysfunctional disorder is one in which T-cells
are anergic or have decreased ability to secrete cytokines,
proliferate, or execute cytolytic activity. In a specific aspect,
the decreased responsiveness results in ineffective control of a
pathogen or tumor expressing an immunogen. Examples of T cell
dysfunctional disorders characterized by T-cell dysfunction include
unresolved acute infection, chronic infection and tumor
immunity.
[0034] "Tumor immunity" refers to the process in which tumors evade
immune recognition and clearance. Thus, as a therapeutic concept,
tumor immunity is "treated" when such evasion is attenuated, and
the tumors are recognized and attacked by the immune system.
Examples of tumor recognition include tumor binding, tumor
shrinkage and tumor clearance.
[0035] "Immunogenicity" refers to the ability of a particular
substance to provoke an immune response. Tumors are immunogenic and
enhancing tumor immunogenicity aids in the clearance of the tumor
cells by the immune response. Examples of enhancing tumor
immunogenicity include treatment with anti-PDL antibodies and a MEK
inhibitor.
[0036] "Sustained response" refers to the sustained effect on
reducing tumor growth after cessation of a treatment. For example,
the tumor size may remain to be the same or smaller as compared to
the size at the beginning of the administration phase. In some
embodiments, the sustained response has a duration at least the
same as the treatment duration, at least 1.5.times., 2.0.times.,
2.5.times., or 3.0.times. length of the treatment duration.
[0037] As used herein, "cancer" and "cancerous" refer to or
describe the physiological condition in mammals that is typically
characterized by unregulated cell growth. Included in this
definition are benign and malignant cancers as well as dormant
tumors or micrometastases. Examples of cancer include but are not
limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia.
More particular examples of such cancers include but are not
limited to squamous cell cancer, lung cancer (including small-cell
lung cancer, non-small cell lung cancer, adenocarcinoma of the
lung, and squamous carcinoma of the lung), cancer of the
peritoneum, hepatocellular cancer, gastric or stomach cancer
(including gastrointestinal cancer), pancreatic cancer,
glioblastoma, cervical cancer, ovarian cancer, liver cancer,
bladder cancer, hepatoma, breast cancer, colon cancer, colorectal
cancer, endometrial or uterine carcinoma, salivary gland carcinoma,
kidney or renal cancer, liver cancer, prostate cancer, vulval
cancer, thyroid cancer, hepatic carcinoma and various types of head
and neck cancer, as well as B-cell lymphoma (including low
grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic
(SL) NHL; intermediate grade/follicular NHL; intermediate grade
diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic
NHL; high grade small non-cleaved cell NHL; bulky disease NHL;
mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's
Macroglobulinemia); chronic lymphocytic leukemia (CLL); acute
lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic
myeloblastic leukemia; and post-transplant lymphoproliferative
disorder (PTLD), as well as abnormal vascular proliferation
associated with phakomatoses, edema (such as that associated with
brain tumors), and Meigs' syndrome. Examples of cancer may include
primary tumors of any of the above types of cancer or metastatic
tumors at a second site derived from any of the above types of
cancer.
[0038] The term "antibody" includes monoclonal antibodies
(including full length antibodies which have an immunoglobulin Lc
region), antibody compositions with polyepitopic specificity,
multispecific antibodies (e.g., bispecific antibodies, diabodies,
and single-chain molecules, as well as antibody fragments (e.g.,
Fab, F(ab').sub.2, and Fv). The term "immunoglobulin" (Ig) is used
interchangeably with "antibody" herein.
[0039] The basic 4-chain antibody unit is a heterotetrameric
glycoprotein composed of two identical light (L) chains and two
identical heavy (H) chains. An IgM antibody consists of 5 of the
basic heterotetramer units along with an additional polypeptide
called a J chain, and contains 10 antigen binding sites, while IgA
antibodies comprise from 2-5 of the basic 4-chain units which can
polymerize to form polyvalent assemblages in combination with the J
chain. In the case of IgGs, the 4-chain unit is generally about
150,000 daltons. Each L chain is linked to an H chain by one
covalent disulfide bond, while the two H chains are linked to each
other by one or more disulfide bonds depending on the H chain
isotype. Each H and L chain also has regularly spaced intrachain
disulfide bridges. Each H chain has at the N-terminus, a variable
domain (V.sub.H) followed by three constant domains (C.sub.H) for
each of the .alpha. and .gamma. chains and four C.sub.H domains for
.mu. and .epsilon. isotypes. Each L chain has at the N-terminus, a
variable domain (V.sub.L) followed by a constant domain at its
other end. The V.sub.L is aligned with the V.sub.H and the C.sub.L
is aligned with the first constant domain of the heavy chain
(C.sub.H1). Particular amino acid residues are believed to form an
interface between the light chain and heavy chain variable domains.
The pairing of a V.sub.H and V.sub.L together forms a single
antigen-binding site. For the structure and properties of the
different classes of antibodies, see e.g., Basic and Clinical
Immunology, 8th Edition, Daniel P. Sties, Abba I. Terr and Tristram
G. Parsolw (eds), Appleton & Lange, Norwalk, Conn., 1994, page
71 and Chapter 6. The L chain from any vertebrate species can be
assigned to one of two clearly distinct types, called kappa and
lambda, based on the amino acid sequences of their constant
domains. Depending on the amino acid sequence of the constant
domain of their heavy chains (CH), immunoglobulins can be assigned
to different classes or isotypes. There are five classes of
immunoglobulins: IgA, IgD, IgE, IgG and IgM, having heavy chains
designated .alpha., .delta., .epsilon., .gamma. and .mu.,
respectively. The .gamma. and .alpha. classes are further divided
into subclasses on the basis of relatively minor differences in the
CH sequence and function, e.g., humans express the following
subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1 and IgA2.
[0040] The "variable region" or "variable domain" of an antibody
refers to the amino-terminal domains of the heavy or light chain of
the antibody. The variable domains of the heavy chain and light
chain may be referred to as "V77" and "V.sub.L", respectively.
These domains are generally the most variable parts of the antibody
(relative to other antibodies of the same class) and contain the
antigen binding sites.
[0041] The term "variable" refers to the fact that certain segments
of the variable domains differ extensively in sequence among
antibodies. The V domain mediates antigen binding and defines the
specificity of a particular antibody for its particular antigen.
However, the variability is not evenly distributed across the
entire span of the variable domains. Instead, it is concentrated in
three segments called hypervariable regions (HVRs) both in the
light-chain and the heavy chain variable domains. The more highly
conserved portions of variable domains are called the framework
regions (FR). The variable domains of native heavy and light chains
each comprise four FR regions, largely adopting a beta-sheet
configuration, connected by three HVRs, which form loops
connecting, and in some cases forming part of, the beta-sheet
structure. The HVRs in each chain are held together in close
proximity by the FR regions and, with the HVRs from the other
chain, contribute to the formation of the antigen binding site of
antibodies (see Kabat et al., Sequences of Immunological Interest,
Fifth Edition, National Institute of Health, Bethesda, Md. (1991)).
The constant domains are not involved directly in the binding of
antibody to an antigen, but exhibit various effector functions,
such as participation of the antibody in antibody-dependent
cellular toxicity.
[0042] 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 and/or post-translation modifications (e.g.,
isomerizations, amidations) that may be present in minor amounts.
Monoclonal antibodies are highly specific, being directed against a
single antigenic site. In contrast to 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 a
variety of techniques, including, for example, the hybridoma method
(e.g., Kohler and Milstein., Nature, 256:495-97 (1975); Hongo et
al., Hybridoma, 14 (3): 253-260 (1995), Harlow et al., Antibodies:
A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2.sup.nd
ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell
Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods
(see, e.g., U.S. Pat. No. 4,816,567), phage-display technologies
(see, e.g., Clackson et al., Nature, 352: 624-628 (1991); Marks et
al., J. Mol. Biol. 222: 581-597 (1992); Sidhu et al., J. Mol. Biol.
338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093
(2004); Fellouse, Proc. Natl. Acad Sci. USA 101(34): 12467-12472
(2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132
(2004), and technologies for producing human or human-like
antibodies in animals that have parts or all of the human
immunoglobulin loci or genes encoding human immunoglobulin
sequences (see, e.g., WO 1998/24893; WO 1996/34096; WO 1996/33735;
WO 1991/10741; Jakobovits et al., Proc. Natl. Acad Sci. USA 90:
2551 (1993); Jakobovits et al., Nature 362: 255-258 (1993);
Bruggemann et al., Year in Immunol. 7:33 (1993); U.S. Pat. Nos.
5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and
5,661,016; Marks et al., Bio/Technology 10: 779-783 (1992); Lonberg
et al., Nature 368: 856-859 (1994); Morrison, Nature 368: 812-813
(1994); Fishwild et al, Nature Biotechnol. 14: 845-851 (1996);
Neuberger, Nature Biotechnol. 14: 826 (1996); and Lonberg and
Huszar, Intern. Rev. Immunol. 13: 65-93 (1995).
[0043] The term "naked antibody" refers to an antibody that is not
conjugated to a cytotoxic moiety or radiolabel.
[0044] The terms "full-length antibody," "intact antibody" or
"whole antibody" are used interchangeably to refer to an antibody
in its substantially intact form, as opposed to an antibody
fragment. Specifically whole antibodies include those with heavy
and light chains including an Fc region. The constant domains may
be native sequence constant domains (e.g., human native sequence
constant domains) or amino acid sequence variants thereof. In some
cases, the intact antibody may have one or more effector
functions.
[0045] An "antibody fragment" comprises a portion of an intact
antibody, preferably the antigen binding and/or the variable region
of the intact antibody. Examples of antibody fragments include Fab,
Fab', F(ab').sub.2 and Fv fragments; diabodies; linear antibodies
(see U.S. Pat. No. 5,641,870, Example 2; Zapata et al, Protein Eng.
8(10): 1057-1062 [1995]); single-chain antibody molecules and
multispecific antibodies formed from antibody fragments. Papain
digestion of antibodies produced two identical antigen-binding
fragments, called "Fab" fragments, and a residual "Fc" fragment, a
designation reflecting the ability to crystallize readily. The Fab
fragment consists of an entire L chain along with the variable
region domain of the H chain (V.sub.H), and the first constant
domain of one heavy chain (C.sub.H1). Each Fab fragment is
monovalent with respect to antigen binding, i.e., it has a single
antigen-binding site. Pepsin treatment of an antibody yields a
single large F(ab').sub.2 fragment which roughly corresponds to two
disulfide linked Fab fragments having different antigen-binding
activity and is still capable of cross-linking antigen. Fab'
fragments differ from Fab fragments by having a few additional
residues at the carboxy terminus of the C.sub.H1 domain including
one or more cysteines from the antibody hinge region. Fab'-SH is
the designation herein for Fab' in which the cysteine residue(s) of
the constant domains bear a free thiol group. F(ab').sub.2 antibody
fragments originally were produced as pairs of Fab' fragments which
have hinge cysteines between them. Other chemical couplings of
antibody fragments are also known.
[0046] The Fc fragment comprises the carboxy-terminal portions of
both H chains held together by disulfides. The effector functions
of antibodies are determined by sequences in the Fc region, the
region which is also recognized by Fc receptors (FcR) found on
certain types of cells.
[0047] "Fv" is the minimum antibody fragment which contains a
complete antigen-recognition and -binding site. This fragment
consists of a dimer of one heavy- and one light-chain variable
region domain in tight, non-covalent association. From the folding
of these two domains emanate six hypervariable loops (3 loops each
from the H and L chain) that contribute the amino acid residues for
antigen binding and confer antigen binding specificity to the
antibody. However, even a single variable domain (or half of an Fv
comprising only three HVRs specific for an antigen) has the ability
to recognize and bind antigen, although at a lower affinity than
the entire binding site.
[0048] "Single-chain Fv" also abbreviated as "sFv" or "scFv" are
antibody fragments that comprise the V.sub.H and V.sub.L antibody
domains connected into a single polypeptide chain. Preferably, the
sFv 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 the sFv, see
Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113,
Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315
(1994).
[0049] "Functional fragments" of the antibodies of the invention
comprise a portion of an intact antibody, generally including the
antigen binding or variable region of the intact antibody or the Fc
region of an antibody which retains or has modified FcR binding
capability. Examples of antibody fragments include linear antibody,
single-chain antibody molecules and multispecific antibodies formed
from antibody fragments.
[0050] The term "diabodies" refers to small antibody fragments
prepared by constructing sFv fragments (see preceding paragraph)
with short linkers (about 5-10) residues) between the V.sub.H and
V.sub.L domains such that inter-chain but not intra-chain pairing
of the V domains is achieved, thereby resulting in a bivalent
fragment, i.e., a fragment having two antigen-binding sites.
Bispecific diabodies are heterodimers of two "crossover" sFv
fragments in which the V.sub.H and V.sub.L domains of the two
antibodies are present on different polypeptide chains. Diabodies
are described in greater detail in, for example, EP 404,097; WO
93/11161; Hollinger el al., Proc. Natl. Acad Sci. USA 90: 6444-6448
(1993).
[0051] 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(are) 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 (U.S. Pat. No. 4,816,567; Morrison
et al., Proc. Natl. Acad Sci. USA, 81:6851-6855 (1984)). Chimeric
antibodies of interest herein include PRIMATIZED.RTM. antibodies
wherein the antigen-binding region of the antibody is derived from
an antibody produced by, e.g., immunizing macaque monkeys with an
antigen of interest. As used herein, "humanized antibody" is used a
subset of "chimeric antibodies."
[0052] "Humanized" forms of non-human (e.g., murine) antibodies are
chimeric antibodies that contain minimal sequence derived from
non-human immunoglobulin. In one embodiment, a humanized antibody
is a human immunoglobulin (recipient antibody) in which residues
from an HVR (hereinafter defined) of the recipient are replaced by
residues from an HVR of a non-human species (donor antibody) such
as mouse, rat, rabbit or non-human primate having the desired
specificity, affinity, and/or capacity. In some instances,
framework ("FR") residues of the human immunoglobulin are replaced
by corresponding non-human residues. Furthermore, humanized
antibodies may comprise residues that are not found in the
recipient antibody or in the donor antibody. These modifications
may be made to further refine antibody performance, such as binding
affinity. In general, a humanized antibody will comprise
substantially all of at least one, and typically two, variable
domains, in which all or substantially all of the hypervariable
loops correspond to those of a non-human immunoglobulin sequence,
and all or substantially all of the FR regions are those of a human
immunoglobulin sequence, although the FR regions may include one or
more individual FR residue substitutions that improve antibody
performance, such as binding affinity, isomerization,
immunogenicity, etc. The number of these amino acid substitutions
in the FR are typically no more than 6 in the H chain, and in the L
chain, no more than 3. The humanized antibody optionally will also
comprise at least a portion of an immunoglobulin constant region
(Fc), typically that of a human immunoglobulin. For further
details, see, e.g., Jones et al., Nature 321:522-525 (1986);
Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op.
Struct. Biol. 2:593-596 (1992). See also, for example, Vaswani and
Hamilton, Am. Allergy, Asthma & Immunol. 1:105-115 (1998);
Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and
Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Pat. Nos.
6,982,321 and 7,087,409.
[0053] A "human antibody" is an antibody that possesses an
amino-acid sequence corresponding to that of an antibody produced
by a human and/or has been made using any of the techniques for
making human antibodies as disclosed herein. This definition of a
human antibody specifically excludes a humanized antibody
comprising non-human antigen-binding residues. Human antibodies can
be produced using various techniques known in the art, including
phage-display libraries. Hoogenboom and Winter, J. Mol. Biol.,
227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Also
available for the preparation of human monoclonal antibodies are
methods described in Cole et al., Monoclonal Antibodies and Cancer
Therapy, Alan R. Liss, p. 77 (1985); Boemer et al., J. Immunol.,
147(1):86-95 (1991). See also van Dijk and van de Winkel, Curr.
Opin. Pharmacol, 5: 368-74 (2001). Human antibodies can be prepared
by administering the antigen to a transgenic animal that has been
modified to produce such antibodies in response to antigenic
challenge, but whose endogenous loci have been disabled, e.g.,
immunized xenomice (see, e.g., U.S. Pat. Nos. 6,075,181 and
6,150,584 regarding XENOMOUSE.TM. technology). See also, for
example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562
(2006) regarding human antibodies generated via a human B-cell
hybridoma technology.
[0054] The term "hypervariable region," "HVR," or "HV," when used
herein refers to the regions of an antibody variable domain which
are hypervariable in sequence and/or form structurally defined
loops. Generally, antibodies comprise six HVRs; three in the VH
(H1, H2, H3), and three in the VL (L1, L2, L3). In native
antibodies, H3 and L3 display the most diversity of the six HVRs,
and H3 in particular is believed to play a unique role in
conferring fine specificity to antibodies. See, e.g., Xu et al.,
Immunity 13:37-45 (2000); Johnson and Wu, in Methods in Molecular
Biology 248:1-25 (Lo, ed., Human Press, Totowa, N.J., 2003).
Indeed, naturally occurring camelid antibodies consisting of a
heavy chain only are functional and stable in the absence of light
chain. See, e.g., Hamers-Casterman et al, Nature 363:446-448
(1993); Sheriff et al., Nature Struct. Biol 3:733-736 (1996).
[0055] A number of HVR delineations are in use and are encompassed
herein. The Kabat Complementarity Determining Regions (CDRs) are
based on sequence variability and are the most commonly used (Kabat
et al., Sequences of Proteins of Immunological Interest, 5th Ed.
Public Health Service, National Institutes of Health, Bethesda, Md.
(1991)). Chothia refers instead to the location of the structural
loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The AbM
HVRs represent a compromise between the Kabat HVRs and Chothia
structural loops, and are used by Oxford Molecular's AbM antibody
modeling software. The "contact" HVRs are based on an analysis of
the available complex crystal structures. The residues from each of
these HVRs are noted below.
TABLE-US-00001 Loop Kabat AbM Chothia Contact L1 L24-L34 L24-L34
L26-L32 L30-L36 L2 L50-L56 L50-L56 L50-L52 L46-L55 L3 L89-L97
L89-L97 L91-L96 L89-L96 H1 H31-H35B H26-H35B H26-H32 H30-H35B
(Kabat numbering) H1 H31-H35 H26-H35 H26-H32 H30-H35 (Chothia
numbering) H2 H50-H65 H50-H58 H53-H55 H47-H58 H3 H95-H102 H95-H102
H96-H101 H93-H101
[0056] HVRs may comprise "extended HVRs" as follows: 24-36 or 24-34
(L1), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3) in the VL and
26-35 (H1), 50-65 or 49-65 (H2) and 93-102, 94-102, or 95-102 (H3)
in the VH. The variable domain residues are numbered according to
Kabat et al., supra, for each of these definitions.
[0057] The expression "variable-domain residue-numbering as in
Kabat" or "amino-acid-position numbering as in Kabat," and
variations thereof, refers to the numbering system used for
heavy-chain variable domains or light-chain variable domains of the
compilation of antibodies in Kabat et al., supra. Using this
numbering system, the actual linear amino acid sequence may contain
fewer or additional amino acids corresponding to a shortening of,
or insertion into, a FR or HVR of the variable domain. For example,
a heavy-chain variable domain may include a single amino acid
insert (residue 52a according to Kabat) after residue 52 of H2 and
inserted residues (e.g. residues 82a, 82b, and 82c, etc. according
to Kabat) after heavy-chain FR residue 82. The Kabat numbering of
residues may be determined for a given antibody by alignment at
regions of homology of the sequence of the antibody with a
"standard" Kabat numbered sequence.
[0058] "Framework" or "FR" residues are those variable-domain
residues other than the HVR residues as herein defined.
[0059] A "human consensus framework" or "acceptor human framework"
is a framework that represents the most commonly occurring amino
acid residues in a selection of human immunoglobulin VL or VH
framework sequences. Generally, the selection of human
immunoglobulin VL or VH sequences is from a subgroup of variable
domain sequences. Generally, the subgroup of sequences is a
subgroup as in Kabat et al., Sequences of Proteins of Immunological
Interest, 5.sup.th Ed. Public Health Service, National Institutes
of Health, Bethesda, Md. (1991). Examples include for the VL, the
subgroup may be subgroup kappa I, kappa II, kappa III or kappa IV
as in Kabat et al., supra. Additionally, for the VH, the subgroup
may be subgroup I, subgroup II, or subgroup III as in Kabat et al.,
supra. Alternatively, a human consensus framework can be derived
from the above in which particular residues, such as when a human
framework residue is selected based on its homology to the donor
framework by aligning the donor framework sequence with a
collection of various human framework sequences. An acceptor human
framework "derived from" a human immunoglobulin framework or a
human consensus framework may comprise the same amino acid sequence
thereof, or it may contain pre-existing amino acid sequence
changes. In some embodiments, the number of pre-existing amino acid
changes are 10 or less, 9 or less, 8 or less, 7 or less, 6 or less,
5 or less, 4 or less, 3 or less, or 2 or less.
[0060] A "VH subgroup III consensus framework" comprises the
consensus sequence obtained from the amino acid sequences in
variable heavy subgroup III of Kabat et al., supra. In one
embodiment, the VH subgroup III consensus framework amino acid
sequence comprises at least a portion or all of each of the
following sequences: EVQLVESGGGLVQPGGSLRLSCAAS (HC-FR1)(SEQ ID
NO:4), WVRQAPGKGLEWV (HC-FR2), (SEQ ID NO:5),
RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (HC-FR3, SEQ ID NO:6), WGQGTLVTVSA
(HC-FR4), (SEQ ID NO:7).
[0061] A "VL kappa I consensus framework" comprises the consensus
sequence obtained from the amino acid sequences in variable light
kappa subgroup I of Kabat et al, supra. In one embodiment, the VH
subgroup I consensus framework amino acid sequence comprises at
least a portion or all of each of the following sequences:
DIQMTQSPSSLSASVGDRVTITC (LC-FR1) (SEQ ID NO: 11), WYQQKPGKAPKLLIY
(LC-FR2) (SEQ ID NO: 12), GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
(LC-FR3)(SEQ ID NO: 13), FGQGTKVEIKR (LC-FR4)(SEQ ID NO: 14).
[0062] An "amino-acid modification" at a specified position, e.g.
of the Fc region, refers to the substitution or deletion of the
specified residue, or the insertion of at least one amino acid
residue adjacent the specified residue. Insertion "adjacent" to a
specified residue means insertion within one to two residues
thereof. The insertion may be N-terminal or C-terminal to the
specified residue. The preferred amino acid modification herein is
a substitution.
[0063] An "affinity-matured" antibody is one with one or more
alterations in one or more HVRs thereof that result in an
improvement in the affinity of the antibody for antigen, compared
to a parent antibody that does not possess those alteration(s). In
one embodiment, an affinity-matured antibody has nanomolar or even
picomolar affinities for the target antigen. Affinity-matured
antibodies are produced by procedures known in the art. For
example, Marks et al., Bio/Technology 10:779-783 (1992) describes
affinity maturation by VH- and VL-domain shuffling. Random
mutagenesis of HVR and/or framework residues is described by, for
example: Barbas et al. Proc Nat. Acad. Sd. USA 91:3809-3813 (1994);
Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol.
155:1994-2004 (1995); Jackson et al, J. Immunol. 154(7):3310-9
(1995); and Hawkins et al, J. Mol. Biol. 226:889-896 (1992).
[0064] As use herein, the term "specifically binds to" or is
"specific for" refers to measurable and reproducible interactions
such as binding between a target and an antibody, which is
determinative of the presence of the target in the presence of a
heterogeneous population of molecules including biological
molecules. For example, an antibody that specifically binds to a
target (which can be an epitope) is an antibody that binds this
target with greater affinity, avidity, more readily, and/or with
greater duration than it binds to other targets. In one embodiment,
the extent of binding of an antibody to an unrelated target is less
than about 10% of the binding of the antibody to the target as
measured, e.g., by a radioimmunoassay (RIA). In certain
embodiments, an antibody that specifically binds to a target has a
dissociation constant (Kd) of .ltoreq.1 .mu.M, .ltoreq.100 nM,
.ltoreq.10 nM, .ltoreq.1 nM, or .ltoreq.0.1 nM. In certain
embodiments, an antibody specifically binds to an epitope on a
protein that is conserved among the protein from different species.
In another embodiment, specific binding can include, but does not
require exclusive binding.
[0065] As used herein, the term "immunoadhesin" designates
antibody-like molecules which combine the binding specificity of a
heterologous protein (an "adhesin") with the effector functions of
immunoglobulin constant domains. Structurally, the immunoadhesins
comprise a fusion of an amino acid sequence with the desired
binding specificity which is other than the antigen recognition and
binding site of an antibody (i.e., is "heterologous"), and an
immunoglobulin constant domain sequence. The adhesin part of an
immunoadhesin molecule typically is a contiguous amino acid
sequence comprising at least the binding site of a receptor or a
ligand. The immunoglobulin constant domain sequence in the
immunoadhesin may be obtained from any immunoglobulin, such as
IgG-1, IgG-2 (including IgG2A and IgG2B), IgG-3, or IgG-4 subtypes,
IgA (including IgA-1 and IgA-2), IgE, IgD or IgM. The Ig fusions
preferably include the substitution of a domain of a polypeptide or
antibody described herein in the place of at least one variable
region within an Ig molecule. In a particularly preferred
embodiment, the immunoglobulin fusion includes the hinge, CH2 and
CH3, or the hinge, CH1, CH2 and CH3 regions of an IgG1 molecule.
For the production of immunoglobulin fusions see also U.S. Pat. No.
5,428,130 issued Jun. 27, 1995. For example, useful immunoadhesins
as second medicaments useful for combination therapy herein include
polypeptides that comprise the extracellular or PD-1 binding
portions of PD-L1 or PD-L2 or the extracellular or PD-L1 or PD-L2
binding portions of PD-1, fused to a constant domain of an
immunoglobulin sequence, such as a PD-L1 ECD-Fc, a PD-L2 ECD-Fc,
and a PD-1 ECD-Fc, respectively. Immunoadhesin combinations of Ig
Fc and ECD of cell surface receptors are sometimes termed soluble
receptors.
[0066] A "fusion protein" and a "fusion polypeptide" refer to a
polypeptide having two portions covalently linked together, where
each of the portions is a polypeptide having a different property.
The property may be a biological property, such as activity in
vitro or in vivo. The property may also be simple chemical or
physical property, such as binding to a target molecule, catalysis
of a reaction, etc. The two portions may be linked directly by a
single peptide bond or through a peptide linker but are in reading
frame with each other.
[0067] A "PD-1 oligopeptide," "PD-L1 oligopeptide," or "PD-L2
oligopeptide" is an oligopeptide that binds, preferably
specifically, to a PD-1, PD-L1 or PD-L2 negative costimulatory
polypeptide, respectively, including a receptor, ligand or
signaling component, respectively, as described herein. Such
oligopeptides may be chemically synthesized using known
oligopeptide synthesis methodology or may be prepared and purified
using recombinant technology. Such oligopeptides are usually at
least about 5 amino acids in length, alternatively at least about
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,
24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,
41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,
58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acids in length or
more. Such oligopeptides may be identified using well known
techniques. In this regard, it is noted that techniques for
screening oligopeptide libraries for oligopeptides that are capable
of specifically binding to a polypeptide target are well known in
the art (see, e.g., U.S. Pat. Nos. 5,556,762, 5,750,373, 4,708,871,
4,833,092, 5,223,409, 5,403,484, 5,571,689, 5,663,143; PCT
Publication Nos. WO 84/03506 and WO84/03564; Geysen et al., Proc.
Natl. Acad Sci. USA., 81:3998-4002 (1984); Geysen et al., Proc
Natl. Acad Sci. USA., 82:178-182 (1985); Geysen et al, in Synthetic
Peptides as Antigens, 130-149 (1986); Geysen et al., J. Immunol.
Meth., 102:259-274 (1987); Schoofs et al, J. Immunol., 140:611-616
(1988), Cwirla, S. E. et al. Proc. Natl. Acad. Sci. USA, 87:6378
(1990); Lowman, H. B. et al. Biochemistry, 30:10832 (1991);
Clackson, T. et al. Nature, 352: 624 (1991); Marks, J. D. et al.,
J. Mol. Biol., 222:581 (1991); Kang, A. S. et al. Proc. Natl. Acad.
ScL USA, 88:8363 (1991), and Smith, G. P., Current Opin.
Biotechnol, 2:668 (1991).
[0068] A "blocking" antibody or an "antagonist" antibody is one
that inhibits or reduces a biological activity of the antigen it
binds. In some embodiments, blocking antibodies or antagonist
antibodies substantially or completely inhibit the biological
activity of the antigen. The anti-PD-L1 antibodies of the invention
block the signaling through PD-1 so as to restore a functional
response by T-cells (e.g., proliferation, cytokine production,
target cell killing) from a dysfunctional state to antigen
stimulation.
[0069] An "agonist" or activating antibody is one that enhances or
initiates signaling by the antigen to which it binds. In some
embodiments, agonist antibodies cause or activate signaling without
the presence of the natural ligand.
[0070] The term "Fc region" herein is used to define a C-terminal
region of an immunoglobulin heavy chain, including native-sequence
Fc regions and variant Fc regions. Although the boundaries of the
Fc region of an immunoglobulin heavy chain might vary, the human
IgG heavy-chain Fc region is usually defined to stretch from an
amino acid residue at position Cys226, or from Pro230, to the
carboxyl-terminus thereof. The C-terminal lysine (residue 447
according to the EU numbering system) of the Fc region may be
removed, for example, during production or purification of the
antibody, or by recombinantly engineering the nucleic acid encoding
a heavy chain of the antibody. Accordingly, a composition of intact
antibodies may comprise antibody populations with all K447 residues
removed, antibody populations with no K447 residues removed, and
antibody populations having a mixture of antibodies with and
without the K447 residue. Suitable native-sequence Fc regions for
use in the antibodies of the invention include human IgG1, IgG2
(IgG2A, IgG2B), IgG3 and IgG4.
[0071] "Fc receptor" or "FcR" describes a receptor that binds to
the Fc region of an antibody. The preferred FcR is a native
sequence human FcR. Moreover, a preferred FcR is one which binds an
IgG antibody (a gamma receptor) and includes receptors of the
Fc.gamma.RI, Fc.gamma.RII, and Fc.gamma.RIII subclasses, including
allelic variants and alternatively spliced forms of these
receptors, Fc.gamma.RII receptors include Fc.gamma.RIIA (an
"activating receptor") and Fc.gamma.RIIB (an "inhibiting
receptor"), which have similar amino acid sequences that differ
primarily in the cytoplasmic domains thereof. Activating receptor
Fc.gamma.RIIA contains an immunoreceptor tyrosine-based activation
motif (ITAM) in its cytoplasmic domain. Inhibiting receptor
Fc.gamma.RIIB contains an immunoreceptor tyrosine-based inhibition
motif (ITIM) in its cytoplasmic domain, (see M. Daeron, Annu. Rev.
Immunol. 15:203-234 (1997). FcRs are reviewed in Ravetch and Kind,
Annu. Rev. Immunol. 9: 457-92 (1991); Capel et al., Immunomethods
4: 25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126: 330-41
(1995). Other FcRs, including those to be identified in the future,
are encompassed by the term "FcR" herein.
[0072] The term "Fc receptor" or "FcR" also includes the neonatal
receptor, FcRn, which is responsible for the transfer of maternal
IgGs to the fetus. Guyer et al., J. Immunol. 117: 587 (1976) and
Kim et al., J. Immunol. 24: 249 (1994). Methods of measuring
binding to FcRn are known (see, e.g., Ghetie and Ward, Immunol.
Today 18: (12): 592-8 (1997); Ghetie et al, Nature Biotechnology 15
(7): 637-40 (1997); Hinton et al., J. Biol. Chem. 279 (8): 6213-6
(2004); WO 2004/92219 (Hinton et al.). Binding to FcRn in vivo and
serum half-life of human FcRn high-affinity binding polypeptides
can be assayed, e.g., in transgenic mice or transfected human cell
lines expressing human FcRn, or in primates to which the
polypeptides having a variant Fc region are administered. WO
2004/42072 (Presta) describes antibody variants which improved or
diminished binding to FcRs. See also, e.g., Shields et al, J. Biol.
Chem. 9(2): 6591-6604 (2001).
[0073] The phrase "substantially reduced," or "substantially
different," as used herein, denotes a sufficiently high degree of
difference between two numeric values (generally one associated
with a molecule and the other associated with a
reference/comparator molecule) such that one of skill in the art
would consider the difference between the two values to be of
statistical significance within the context of the biological
characteristic measured by said values (e.g., Kd values). The
difference between said two values is, for example, greater than
about 10%, greater than about 20%, greater than about 30%, greater
than about 40%, and/or greater than about 50% as a function of the
value for the reference/comparator molecule.
[0074] The term "substantially similar" or "substantially the
same," as used herein, denotes a sufficiently high degree of
similarity between two numeric values (for example, one associated
with an antibody of the invention and the other associated with a
reference/comparator antibody), such that one of skill in the art
would consider the difference between the two values to be of
little or no biological and/or statistical significance within the
context of the biological characteristic measured by said values
(e.g., Kd values). The difference between said two values is, for
example, less than about 50%, less than about 40%, less than about
30%, less than about 20%, and/or less than about 10% as a function
of the reference/comparator value.
[0075] "Carriers" as used herein include pharmaceutically
acceptable carriers, excipients, or stabilizers that are nontoxic
to the cell or mammal being exposed thereto at the dosages and
concentrations employed. Often the physiologically acceptable
carrier is an aqueous pH buffered solution. Examples of
physiologically acceptable carriers include buffers such as
phosphate, citrate, and other organic acids; antioxidants including
ascorbic acid; low molecular weight (less than about 10 residues)
polypeptide; proteins, such as serum albumin, gelatin, or
immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone;
amino acids such as glycine, glutamine, asparagine, arginine or
lysine; monosaccharides, disaccharides, and other carbohydrates
including glucose, mannose, or dextrins; chelating agents such as
EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming
counterions such as sodium; and/or nonionic surfactants such as
TWEEN.TM., polyethylene glycol (PEG), and PLURONICS.TM..
[0076] A "package insert" refers to instructions customarily
included in commercial packages of medicaments that contain
information about the indications customarily included in
commercial packages of medicaments that contain information about
the indications, usage, dosage, administration, contraindications,
other medicaments to be combined with the packaged product, and/or
warnings concerning the use of such medicaments, etc.
[0077] As used herein, the term "treatment" refers to clinical
intervention designed to alter the natural course of the individual
or cell being treated during the course of clinical pathology.
Desirable effects of treatment include decreasing the rate of
disease progression, ameliorating or palliating the disease state,
and remission or improved prognosis. For example, an individual is
successfully "treated" if one or more symptoms associated with
cancer are mitigated or eliminated, including, but are not limited
to, reducing the proliferation of (or destroying) cancerous cells,
decreasing symptoms resulting from the disease, increasing the
quality of life of those suffering from the disease, decreasing the
dose of other medications required to treat the disease, delaying
the progression of the disease, and/or prolonging survival of
individuals.
[0078] As used herein, "delaying progression of a disease" means to
defer, hinder, slow, retard, stabilize, and/or postpone development
of the disease (such as cancer). This delay can be of varying
lengths of time, depending on the history of the disease and/or
individual being treated. As is evident to one skilled in the art,
a sufficient or significant delay can, in effect, encompass
prevention, in that the individual does not develop the disease.
For example, a late stage cancer, such as development of
metastasis, may be delayed.
[0079] As used herein, "reducing or inhibiting cancer relapse"
means to reduce or inhibit tumor or cancer relapse or tumor or
cancer progression. As disclosed herein, cancer relapse and/or
cancer progression include, without limitation, cancer
metastasis.
[0080] An "effective amount" is at least the minimum concentration
required to effect a measurable improvement or prevention of a
particular disorder. An effective amount herein may vary according
to factors such as the disease state, age, sex, and weight of the
patient, and the ability of the antibody to elicit a desired
response in the individual. An effective amount is also one in
which any toxic or detrimental effects of the treatment are
outweighed by the therapeutically beneficial effects. For
prophylactic use, beneficial or desired results include results
such as eliminating or reducing the risk, lessening the severity,
or delaying the onset of the disease, including biochemical,
histological and/or behavioral symptoms of the disease, its
complications and intermediate pathological phenotypes presenting
during development of the disease. For therapeutic use, beneficial
or desired results include clinical results such as decreasing one
or more symptoms resulting from the disease, increasing the quality
of life of those suffering from the disease, decreasing the dose of
other medications required to treat the disease, enhancing effect
of another medication such as via targeting, delaying the
progression of the disease, and/or prolonging survival. In the case
of cancer or tumor, an effective amount of the drug may have the
effect in reducing the number of cancer cells; reducing the tumor
size; inhibiting (i.e., slow to some extent or desirably stop)
cancer cell infiltration into peripheral organs; inhibit (i.e.,
slow to some extent and desirably stop) tumor metastasis;
inhibiting to some extent tumor growth; and/or relieving to some
extent one or more of the symptoms associated with the disorder. An
effective amount can be administered in one or more
administrations. For purposes of this invention, an effective
amount of drug, compound, or pharmaceutical composition is an
amount sufficient to accomplish prophylactic or therapeutic
treatment either directly or indirectly. As is understood in the
clinical context, an effective amount of a drug, compound, or
pharmaceutical composition may or may not be achieved in
conjunction with another drug, compound, or pharmaceutical
composition. Thus, an "effective amount" may be considered in the
context of administering one or more therapeutic agents, and a
single agent may be considered to be given in an effective amount
if, in conjunction with one or more other agents, a desirable
result may be or is achieved.
[0081] As used herein, "in conjunction with" refers to
administration of one treatment modality in addition to another
treatment modality. As such, "in conjunction with" refers to
administration of one treatment modality before, during, or after
administration of the other treatment modality to the
individual.
[0082] As used herein, "complete response" or "CR" refers to
disappearance of all target lesions; "partial response" or "PR"
refers to at least a 30% decrease in the sum of the longest
diameters (SLD) of target lesions, taking as reference the baseline
SLD; and "stable disease" or "SD" refers to neither sufficient
shrinkage of target lesions to qualify for PR, nor sufficient
increase to qualify for PD, taking as reference the smallest SLD
since the treatment started.
[0083] As used herein, "progressive disease" or "PD" refers to at
least a 20% increase in the SLD of target lesions, taking as
reference the smallest SLD recorded since the treatment started or
the presence of one or more new lesions.
[0084] As used herein, "progression free survival" (PFS) refers to
the length of time during and after treatment during which the
disease being treated (e.g., cancer) does not get worse.
Progression-free survival may include the amount of time patients
have experienced a complete response or a partial response, as well
as the amount of time patients have experienced stable disease.
[0085] As used herein, "overall response rate" (ORR) refers to the
sum of complete response (CR) rate and partial response (PR)
rate.
[0086] As used herein, "overall survival" refers to the percentage
of individuals in a group who are likely to be alive after a
particular duration of time.
[0087] A "chemotherapeutic agent" is a chemical compound useful in
the treatment of cancer. Examples of chemotherapeutic agents
include alkylating agents such as thiotepa and cyclophosphamide
(CYTOXAN.RTM.); alkyl sulfonates such as busulfan, improsulfan, and
piposulfan; aziridines such as benzodopa, carboquone, meturedopa,
and uredopa; ethylenimines and methylamelamines including
altretamine, triethylenemelamine, trietylenephosphoramide,
triethiylenethiophosphoramide and trimethylolomelamine; acetogenins
(especially bullatacin and bullatacinone);
delta-9-tetrahydrocannabinol (dronabinol, MARINOL.RTM.);
beta-lapachone; lapachol; colchicines; betulinic acid; a
camptothecin (including the synthetic analogue topotecan
(HYCAMTIN.RTM.), CPT-11 (irinotecan, CAMPTOSAR.RTM.),
acetylcamptothecin, scopolectin, and 9-aminocamptothecin);
bryostatin; pemetrexed; callystatin; CC-1065 (including its
adozelesin, carzelesin and bizelesin synthetic analogues);
podophyllotoxin; podophyllinic acid; teniposide; cryptophycins
(particularly cryptophycin 1 and cryptophycin 8); dolastatin;
duocarmycin (including the synthetic analogues, KW-2189 and
CB1-TM1); eleutherobin; pancratistatin; TLK-286; CDP323, an oral
alpha-4 integrin inhibitor; a sarcodictyin; spongistatin; nitrogen
mustards such as chlorambucil, chlornaphazine, cholophosphamide,
estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide
hydrochloride, melphalan, novembichin, phenesterine, prednimustine,
trofosfamide, uracil mustard; nitrosureas such as carmustine,
chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine;
antibiotics such as the enediyne antibiotics (e.g., calicheamicin,
especially calicheamicin gamma1I and calicheamicin omegaI1 (see,
e.g., Nicolaou et al., Angew. Chem Intl. Ed Engl., 33: 183-186
(1994)); dynemicin, including dynemicin A; an esperamicin; as well
as neocarzinostatin chromophore and related chromoprotein enediyne
antibiotic chromophores), aclacinomysins, actinomycin, authramycin,
azaserine, bleomycins, cactinomycin, carabicin, carminomycin,
carzinophilin, chromomycinis, dactinomycin, daunorubicin,
detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including
ADRIAMYCIN.RTM., morpholino-doxombicin, cyanomorpholino-doxombicin,
2-pyrrolino-doxorubicin, doxorubicin HCl liposome injection
(DOXIL.RTM.) and deoxydoxorubicin), epirubicin, esombicin,
idarubicin, marcellomycin, mitomycins such as mitomycin C,
mycophenolic acid, nogalamycin, olivomycins, peplomycin,
potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin,
streptozocin, tubercidin, ubenimex, zinostatin, zorubicin;
anti-metabolites such as methotrexate, gemcitabine (GEMZAR.RTM.),
tegafur (UFTORAL.RTM.), capecitabine (XELODA.RTM.), an epothilone,
and 5-fluorouracil (5-FU); folic acid analogues such as denopterin,
methotrexate, pteropterin, trimetrexate; purine analogs such as
fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine
analogs such as ancitabine, azacitidine, 6-azauridine, carmofur,
cytarabine, dideoxyuridine, doxifluridine, enocitabine,
floxuridine, and imatinib (a 2-phenylaminopyrimidine derivative),
as well as other c-Kit inhibitors; anti-adrenals such as
aminoglutethimide, mitotane, trilostane; folic acid replenisher
such as frolinic acid; aceglatone; aldophosphamide glycoside;
aminolevulinic acid; eniluracil; amsacrine; bestrabucil;
bisantrene; edatraxate; defofamine; demecolcine; diaziquone;
elfornithine; elliptinium acetate; etoglucid; gallium nitrate;
hydroxyurea; lentinan; lonidainine; maytansinoids such as
maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol;
nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone;
2-ethylhydrazide; procarbazine; PSK.RTM. polysaccharide complex
(JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin;
sizofiran; spirogermanium; tenuazonic acid; triaziquone;
2,2',2''-trichlorotriethylamine; trichothecenes (especially T-2
toxin, verracurin A, roridin A and anguidine); urethan; vindesine
(ELDISINE.RTM., FILDESIN.RTM.); dacarbazine; mannomustine;
mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside
("Ara-C"); thiotepa; taxoids, e.g., paclitaxel (TAXOL.RTM.),
albumin-engineered nanoparticle formulation of paclitaxel
(ABRAXANE.TM.), and doxetaxel (TAXOTERE.RTM.); chloranbucil;
6-thioguanine; mercaptopurine; methotrexate; platinum analogs such
as cisplatin and carboplatin; vinblastine (VELBAN.RTM.); platinum;
etoposide (VP-16); ifosfamide; mitoxantrone; vincristine
(ONCOVIN.RTM.); oxaliplatin; leucovovin; vinorelbine
(NAVELBINE.RTM.); novantrone; edatrexate; daunomycin; aminopterin;
ibandronate; topoisomerase inhibitor RFS 2000;
difluorometlhylornithine (DMFO); retinoids such as retinoic acid;
pharmaceutically acceptable salts, acids or derivatives of any of
the above; as well as combinations of two or more of the above such
as CHOP, an abbreviation for a combined therapy of
cyclophosphamide, doxorubicin, vincristine, and prednisolone, and
FOLFOX, an abbreviation for a treatment regimen with oxaliplatin
(ELOXATIN.TM.) combined with 5-FU and leucovovin.
[0088] Additional examples of chemotherapeutic agents include
anti-hormonal agents that act to regulate, reduce, block, or
inhibit the effects of hormones that can promote the growth of
cancer, and are often in the form of systemic, or whole-body
treatment. They may be hormones themselves. Examples include
anti-estrogens and selective estrogen receptor modulators (SERMs),
including, for example, tamoxifen (including NOLVADEX.RTM.
tamoxifen), raloxifene (EVISTA.RTM.), droloxifene,
4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone,
and toremifene (FARESTON.RTM.); anti-progesterones; estrogen
receptor down-regulators (ERDs); estrogen receptor antagonists such
as fulvestrant (FASLODEX.RTM.); agents that function to suppress or
shut down the ovaries, for example, leutinizing hormone-releasing
hormone (LHRH) agonists such as leuprolide acetate (LUPRON.RTM. and
ELIGARD.RTM.), goserelin acetate, buserelin acetate and
tripterelin; anti-androgens such as flutamide, nilutamide and
bicalutamide; and aromatase inhibitors that inhibit the enzyme
aromatase, which regulates estrogen production in the adrenal
glands, such as, for example, 4(5)-imidazoles, aminoglutethimide,
megestrol acetate (MEGASE.RTM.), exemestane (AROMASIN.RTM.),
formestanie, fadrozole, vorozole (RIVISOR.RTM.), letrozole
(FEMARA.RTM.), and anastrozole (ARIMIDEX.RTM.). In addition, such
definition of chemotherapeutic agents includes bisphosphonates such
as clodronate (for example, BONEFOS.RTM. or OSTAC.RTM.), etidronate
(DIDROCAL.RTM.), NE-58095, zoledronic acid/zoledronate
(ZOMETA.RTM.), alendronate (FOSAMAX.RTM.), pamidronate
(AREDIA.RTM.), tiludronate (SKELID.RTM.), or risedronate
(ACTONEL.RTM.); as well as troxacitabine (a 1,3-dioxolane
nucleoside cytosine analog); anti-sense oligonucleotides,
particularly those that inhibit expression of genes in signaling
pathways implicated in abherant cell proliferation, such as, for
example, PKC-alpha, Raf, H-Ras, and epidermal growth factor
receptor (EGF-R); vaccines such as THERATOPE.RTM. vaccine and gene
therapy vaccines, for example, ALLOVECTIN.RTM. vaccine,
LEUVECTIN.RTM. vaccine, and VAXID.RTM. vaccine; topoisomerase 1
inhibitor (e.g., LURTOTECAN.RTM.); an anti-estrogen such as
fulvestrant; a Kit inhibitor such as imatinib or EXEL-0862 (a
tyrosine kinase inhibitor); EGFR inhibitor such as erlotinib or
cetuximab; an anti-VEGF inhibitor such as bevacizumab; arinotecan;
rmRH (e.g., ABARELIX.RTM.); lapatinib and lapatinib ditosylate (an
ErbB-2 and EGFR dual tyrosine kinase small-molecule inhibitor also
known as GW572016); 17AAG (geldanamycin derivative that is a heat
shock protein (Hsp) 90 poison), and pharmaceutically acceptable
salts, acids or derivatives of any of the above.
[0089] As used herein, the term "cytokine" refers generically to
proteins released by one cell population that act on another cell
as intercellular mediators or have an autocrine effect on the cells
producing the proteins. Examples of such cytokines include
lymphokines, monokines; interleukins ("ILs") such as IL-1, IL-1a,
IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL10, IL-11, IL-12,
IL-13, IL-15, IL-17A-F, IL-18 to IL-29 (such as IL-23), IL-31,
including PROLEUKIN.RTM. rIL-2; a tumor-necrosis factor such as
TNF-.alpha. or TNF-.beta., TGF-.beta.1-3; and other polypeptide
factors including leukemia inhibitory factor ("LIF"), ciliary
neurotrophic factor ("CNTF"), CNTF-like cytokine ("CLC"),
cardiotrophin ("CT"), and kit ligand ("KL").
[0090] As used herein, the term "chemokine" refers to soluble
factors (e.g., cytokines) that have the ability to selectively
induce chemotaxis and activation of leukocytes. They also trigger
processes of angiogenesis, inflammation, wound healing, and
tumorigenesis. Example chemokines include IL-8, a human homolog of
murine keratinocyte chemoattractant (KC).
[0091] As used herein and in the appended claims, the singular
forms "a," "or," and "the" include plural referents unless the
context clearly dictates otherwise.
[0092] Reference to "about" a value or parameter herein includes
(and describes) variations that are directed to that value or
parameter per se. For example, description referring to "about X"
includes description of "X".
[0093] The term "alkyl" as used herein refers to a saturated linear
or branched-chain monovalent hydrocarbon radical of one to twelve
carbon atoms. Examples of alkyl groups include, but are not limited
to, methyl (Me, --CH.sub.3), ethyl (Et, --CH.sub.2CH.sub.3),
1-propyl (n-Pr, n-propyl, --CH.sub.2CH.sub.2CH.sub.3), 2-propyl
(i-Pr, i-propyl, --CH(CH.sub.3).sub.2), 1-butyl (n-Bu, n-butyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propyl (i-Bu,
i-butyl, --CH.sub.2CH(CH.sub.3).sub.2), 2-butyl (s-Bu, s-butyl,
--CH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propyl (t-Bu, t-butyl,
--C(CH.sub.3).sub.3), 1-pentyl (n-pentyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentyl
(--CH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butyl
(--C(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butyl
(--CH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butyl
(--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butyl
(--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 3-hexyl
(--CH(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)),
2-methyl-2-pentyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.2CH.sub.3),
3-methyl-2-pentyl (--CH(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3),
4-methyl-2-pentyl (--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2),
3-methyl-3-pentyl (--C(CH.sub.3)(CH.sub.2CH.sub.3).sub.2),
2-methyl-3-pentyl (--CH(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2),
2,3-dimethyl-2-butyl (--C(CH.sub.3).sub.2CH(CH.sub.3).sub.2),
3,3-dimethyl-2-butyl (--CH(CH.sub.3)C(CH.sub.3).sub.3, 1-heptyl,
1-octyl, and the like.
[0094] The term "alkenyl" refers to linear or branched-chain
monovalent hydrocarbon radical of two to twelve carbon atoms with
at least one site of unsaturation, i.e., a carbon-carbon, sp.sup.2
double bond, wherein the alkenyl radical includes radicals having
"cis" and "trans" orientations, or alternatively, "E" and "Z"
orientations. Examples include, but are not limited to, ethylenyl
or vinyl (--CH.dbd.CH.sub.2), allyl (--CH.sub.2CH.dbd.CH.sub.2),
and the like.
[0095] The term "alkynyl" refers to a linear or branched monovalent
hydrocarbon radical of two to twelve carbon atoms with at least one
site of unsaturation, i.e., a carbon-carbon, sp triple bond.
Examples include, but are not limited to, ethynyl (--C.ident.CH),
propynyl (propargyl, --CH.sub.2C.ident.CH), and the like.
[0096] The terms "carbocycle", "carbocyclyl", "carbocyclic ring"
and "cycloalkyl" refer to a monovalent non-aromatic, saturated or
partially unsaturated ring having 3 to 12 carbon atoms as a
monocyclic ring or 7 to 12 carbon atoms as a bicyclic ring.
Bicyclic carbocycles having 7 to 12 atoms can be arranged, for
example, as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, and
bicyclic carbocycles having 9 or 10 ring atoms can be arranged as a
bicyclo [5,6] or [6,6] system, or as bridged systems such as
bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane and
bicyclo[3.2.2]nonane. Examples of monocyclic carbocycles include,
but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,
1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl,
cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl,
1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl,
cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, and the
like.
[0097] "Aryl" means a monovalent aromatic hydrocarbon radical of
6-18 carbon atoms derived by the removal of one hydrogen atom from
a single carbon atom of a parent aromatic ring system. Some aryl
groups are represented in the exemplary structures as "Ar". Aryl
includes bicyclic radicals comprising an aromatic ring fused to a
saturated, partially unsaturated ring, or aromatic carbocyclic or
heterocyclic ring. Typical aryl groups include, but are not limited
to, radicals derived from benzene (phenyl), substituted benzenes,
naphthalene, anthracene, indenyl, indanyl, 1,2-dihydronaphthalene,
1,2,3,4-tetrahydronaphthyl, and the like.
[0098] The terms "heterocycle," "heterocyclyl" and "heterocyclic
ring" are used interchangeably herein and refer to a saturated or a
partially unsaturated (i.e., having one or more double and/or
triple bonds within the ring) carbocyclic radical of 3 to 18 ring
atoms in which at least one ring atom is a heteroatom selected from
nitrogen, oxygen and sulfur, the remaining ring atoms being C,
where one or more ring atoms is optionally substituted
independently with one or more substituents described below. A
heterocycle may be a monocycle having 3 to 7 ring members (2 to 6
carbon atoms and 1 to 4 heteroatoms selected from N, O, P, and S)
or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and 1
to 6 heteroatoms selected from N, O, P, and S), for example: a
bicyclo [4,5], [5,5], [5,6], or [6,6] system. Heterocycles are
described in Paquette, Leo A.; "Principles of Modern Heterocyclic
Chemistry" (W. A. Benjamin, New York, 1968), particularly Chapters
1, 3, 4, 6, 7, and 9; "The Chemistry of Heterocyclic Compounds, A
series of Monographs" (John Wiley & Sons, New York, 1950 to
present), in particular Volumes 13, 14, 16, 19, and 28; and J. Am.
Chem. Soc. (1960) 82:5566. "Heterocyclyl" also includes radicals
where heterocycle radicals are fused with a saturated, partially
unsaturated ring, or aromatic carbocyclic or heterocyclic ring.
Examples of heterocyclic rings include, but are not limited to,
pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl,
tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl,
piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, piperazinyl,
homopiperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl,
oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl,
2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl,
dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl,
dihydropyranyl, dihydrothienyl, dihydrofuranyl,
pyrazolidinylimidazolinyl, imidazolidinyl,
3-azabicyco[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, and
azabicyclo[2.2.2]hexanyl. Spiro moieties are also included within
the scope of this definition. Examples of a heterocyclic group
wherein ring atoms are substituted with oxo (.dbd.O) moieties are
pyrimidinonyl and 1,1-dioxo-thiomorpholinyl.
[0099] The term "heteroaryl" refers to a monovalent aromatic
radical of 5- or 6-membered rings, and includes fused ring systems
(at least one of which is aromatic) of 5-18 atoms, containing one
or more heteroatoms independently selected from nitrogen, oxygen,
and sulfur. Examples of heteroaryl groups are pyridinyl (including,
for example, 2-hydroxypyridinyl), imidazolyl, imidazopyridinyl,
pyrimidinyl (including, for example, 4-hydroxypyrimidinyl),
pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl,
isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl,
quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl,
cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,
triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, triazolyl,
thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,
benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,
naphthyridinyl, and furopyridinyl.
[0100] The heterocycle or heteroaryl groups may be carbon
(carbon-linked) or nitrogen (nitrogen-linked) attached where such
is possible. By way of example and not limitation, carbon bonded
heterocycles or heteroaryls are bonded at position 2, 3, 4, 5, or 6
of a pyridine, position 3, 4, 5, or 6 of a pyridazine, position 2,
4, 5, or 6 of a pyrimidine, position 2, 3, 5, or 6 of a pyrazine,
position 2, 3, 4, or 5 of a furan, tetrahydrofuran, thiofuran,
thiophene, pyrrole or tetrahydropyrrole, position 2, 4, or 5 of an
oxazole, imidazole or thiazole, position 3, 4, or 5 of an
isoxazole, pyrazole, or isothiazole, position 2 or 3 of an
aziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4,
5, 6, 7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of
an isoquinoline.
[0101] By way of example and not limitation, nitrogen bonded
heterocycles or heteroaryls are bonded at position 1 of an
aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline,
3-pyrroline, imidazole, imidazolidine, 2-imidazoline,
3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline,
piperidine, piperazine, indole, indoline, 1H-indazole, position 2
of a isoindole, or isoindoline, position 4 of a morpholine, and
position 9 of a carbazole, or .beta.-carboline.
[0102] The heteroatoms present in heteroaryl or heterocyclcyl
include the oxidized forms such as N.sup.+.fwdarw.O.sup.-, S(O) and
S(O).sub.2.
[0103] The term "halo" refers to F, Cl, Br or I.
[0104] The phrase "pharmaceutically acceptable salt" as used
herein, refers to pharmaceutically acceptable organic or inorganic
salts of a compound of the invention. Exemplary salts include, but
are not limited, to sulfate, citrate, acetate, oxalate, chloride,
bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate,
isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate,
tannate, pantothenate, bitartrate, ascorbate, succinate, maleate,
gentisinate, fumarate, gluconate, glucuronate, saccharate, formate,
benzoate, glutamate, methanesulfonate "mesylate", ethanesulfonate,
benzenesulfonate, p-toluenesulfonate, pamoate (i.e.,
1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts, alkali metal
(e.g., sodium and potassium) salts, alkaline earth metal (e.g.,
magnesium) salts, and ammonium salts. A pharmaceutically acceptable
salt may involve the inclusion of another molecule such as an
acetate ion, a succinate ion or other counter ion. The counter ion
may be any organic or inorganic moiety that stabilizes the charge
on the parent compound. Furthermore, a pharmaceutically acceptable
salt may have more than one charged atom in its structure.
Instances where multiple charged atoms are part of the
pharmaceutically acceptable salt can have multiple counter ions.
Hence, a pharmaceutically acceptable salt can have one or more
charged atoms and/or one or more counter ion.
[0105] If the compound of the invention is a base, the desired
pharmaceutically acceptable salt may be prepared by any suitable
method available in the art, for example, treatment of the free
base with an inorganic acid, such as hydrochloric acid, hydrobromic
acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric
acid and the like, or with an organic acid, such as acetic acid,
maleic acid, succinic acid, mandelic acid, fumaric acid, malonic
acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a
pyranosidyl acid, such as glucuronic acid or galacturonic acid, an
alpha hydroxy acid, such as citric acid or tartaric acid, an amino
acid, such as aspartic acid or glutamic acid, an aromatic acid,
such as benzoic acid or cinnamic acid, a sulfonic acid, such as
p-toluenesulfonic acid or ethanesulfonic acid, or the like.
[0106] If the compound of the invention is an acid, the desired
pharmaceutically acceptable salt may be prepared by any suitable
method, for example, treatment of the free acid with an inorganic
or organic base, such as an amine (primary, secondary or tertiary),
an alkali metal hydroxide or alkaline earth metal hydroxide, or the
like. Illustrative examples of suitable salts include, but are not
limited to, organic salts derived from amino acids, such as glycine
and arginine, ammonia, primary, secondary, and tertiary amines, and
cyclic amines, such as piperidine, morpholine and piperazine, and
inorganic salts derived from sodium, calcium, potassium, magnesium,
manganese, iron, copper, zinc, aluminum and lithium.
[0107] The phrase "pharmaceutically acceptable" indicates that the
substance or composition must be compatible chemically and/or
toxicologically, with the other ingredients comprising a
formulation, and/or the mammal being treated therewith.
[0108] A"solvate" refers to an association or complex of one or
more solvent molecules and a compound of the invention. Examples of
solvents that form solvates include, but are not limited to, water,
isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid,
and ethanolamine. The term "hydrate" refers to the complex where
the solvent molecule is water.
[0109] It is understood that aspects and variations of the
invention described herein include "consisting of" and/or
"consisting essentially of" aspects and variations.
III. Methods
[0110] In one aspect, provided herein is a method for treating or
delaying progression of cancer in an individual comprising
administering to the individual an effective amount of a PD-1 axis
binding antagonist and a MEK inhibitor.
[0111] The methods of this invention may find use in treating
conditions where enhanced immunogenicity is desired such as
increasing tumor immunogenicity for the treatment of cancer. A
variety of cancers may be treated, or their progression may be
delayed, including but are not limited to a cancer that may contain
a B-raf V600E mutation, a cancer that may contain a B-raf wildtype,
a cancer that may contain a KRAS wildtype, or a cancer that may
contain an activating KRAS mutation.
[0112] In some embodiments, the individual has cancer or is at risk
of developing cancer that is resistant to a B-raf antagonist. In
some embodiments, the individual has been previously treated with a
B-raf antagonist for cancer. In some embodiments, the individual
has not been previously treated with a B-raf antagonist. B-raf is a
serine-threonine kinase known to be frequently mutated in cancer,
e.g., malignant melanoma, colorectal, ovarian, and thyroid cancer.
Typically, B-raf mutations observed in cancer include activating
mutations, such as the V600E mutation. Without wishing to be bound
to theory, it is thought that activating mutations in B-raf promote
deregulated MAPK/ERK signaling, leading to tumor cell proliferation
and survival.
[0113] While B-raf antagonists (e.g., B-raf inhibitors) are known
to produce effective short-term increases in patient survival and
tumor regression, resistance to B-raf inhibition is frequently
observed (see, e.g., Tentori, L., et al. Trends Pharmacol. Sci.
34(12):656-66 (2013)). Resistance to B-raf inhibition may be
characterized by numerous phenomena. In some embodiments,
resistance to B-raf inhibition may be characterized by one or more
of MAPK pathway reactivation, PI3K activation, CRAF upregulation,
NRAS mutation, PDGFR overexpression, COT1 overexpression, IGFR-1
overexpression, MEK1 mutation, HGF expression, PD-L1
overexpression, MEK2 mutation, MITF focal amplification, AKT
mutation (e.g., AKT1 or AKT3), B-raf amplification, and the
formation of RAF dimers (e.g., CRAF/CRAF dimers, CRAF/B-raf dimers,
and mutated dimerizing B-raf such as a B-raf variant lacking exons
4-8).
[0114] In some embodiments, resistance to B-raf inhibition may
refer to a cancer cell or tumor that is refractory to B-raf
inhibition. Resistance to B-raf inhibition is used herein in the
broadest sense and may include any cancer cell that was previously
or may be expected to be sensitive to B-raf inhibition through any
particular mechanism or at any particular dose of a B-raf
antagonist of the present disclosure. Resistance to B-raf
inhibition may refer to B-raf activity in the presence of a B-raf
antagonist. Resistance to B-raf inhibition may refer to a cell that
grows in the presence of a B-raf antagonist when it is not expected
to grow under such a condition, regardless of the enzymatic
activity of B-raf that may be present.
[0115] In some embodiments, the B-raf antagonist is a small
molecule inhibitor, an antibody, a peptide, a peptidomimetic, an
aptamer or a polynucleotide.
[0116] In some embodiments, the individual has previously been
treated with a B-raf antagonist. In some embodiments, a B-raf
antagonist may include vemurafenib (also known as ZELBORAF.RTM.),
dabrafenib (also known as TAFINLAR.RTM.), LGX818, GSK 2118436,
RAF265, XL281, ARQ736, BAY73-4506, sorafenib, PLX4720, PLX-3603,
GSK2118436, GDC-0879, or
N-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluo-
rophenyl)propane-1-sulfonamide. In some embodiments, a B-raf
antagonist may include MLN2480, LY3009120, a MEK inhibitor such as
trametinib (also known as MEKINIST.RTM.), or an EGFR inhibitor such
as erlotinib (also known as TARCEVA.RTM.). Further descriptions of
B-raf inhibitors may be found in Zambon, A. et al. Bioorg. Med.
Chem. Lett. 22(2):789-92 (2012); Tentori, L., et al. Trends
Pharmacol. Sci. 34(12):656-66 (2013); and Martin-Liberal, J. and
Larkin, J. Expert Opin. Pharmacother. 15(9): 1235-45 (2014),
WO2007/002325, WO2007/002433, WO2009111278, WO2009111279,
WO2009111277, WO2009111280 and U.S. Pat. No. 7,491,829.
[0117] In some embodiments, the B-raf antagonist is a selective
B-raf antagonist of B-raf V600. In some embodiments, the selective
B-raf antagonist of B-raf V600 is a selective antagonist of B-raf
V600E. In some embodiments, the selective B-raf antagonist of B-raf
V600 is a selective antagonist of B-raf V600E, B-raf V600K, and/or
V600D. In some embodiments, the selective B-raf antagonist of B-raf
V600 is a selective antagonist of B-raf V600R. Techniques for
generating and assaying B-raf antagonists that are selective for
B-raf V600 have been described in the art; see, e.g., Tsai, J, et
al., Proc. Natl. Acad. Sci. 105(8):3041-6 (2008).
[0118] In some embodiments, the cancer contains a BRAF V600E
mutation, a BRAF wildtype, a KRAS wildtype, or an activating KRAS
mutation. Methods for detecting the presence of such mutations may
include, without limitation, PCR, Sanger sequencing, use of a
mutation-specific antibody, and the like. Methods for determining
whether a cancer expresses a B-raf V600 are known in the art,
including without limitation the COBAS.RTM. 4800 B-raf V600
Mutation Test kit (Roche).
[0119] In some embodiments, the patient's cancer has been shown to
express a B-raf biomarker. In some embodiments, B-raf biomarker is
mutant B-raf. In some embodiments, mutant B-raf is B-raf V600. In
some embodiments, B-raf V600 is B-raf V600E. In some embodiments,
mutant B-raf is constitutively active.
[0120] In some embodiments, the cancer patient has progressed while
receiving a B-raf antagonist therapy (i.e., the patient is "B-raf
refractory"), or the patient has progressed within 1 month, 2
months, 3 months, 4 months, 5, months, 6 months, 7 months, 8
months, 9 months, 10 months, 11, months, 12 months, or more after
completing a B-raf antagonist-based therapy regimen.
[0121] In some embodiments, vemurafenib resistant cancer is meant
that the cancer patient has progressed while receiving
vemurafenib-based therapy (i.e., the patient is "vemurafenib
refractory"). In some embodiments, cancer in the patient has
progressed within 1 month, 2 months, 3 months, 4 months, 5, months,
6 months, 7 months, 8 months, 9 months, 10 months, 11, months, 12
months, or more after completing a vemurafenib-based therapy
regimen. In some embodiments, the cancer in the individual has
progressed within 1 month, 2 months, 3 months, 4 months, 5, months,
6 months, 7 months, 8 months, 9 months, 10 months, 11, months, 1
year, 2 years, 3 years, 4 years, or 5 years after completing a
B-raf antagonist-based therapy regimen.
[0122] In some embodiments, the treatment results in a sustained
response in the individual after cessation of the treatment.
[0123] In some embodiments, resistance to, e.g., B-raf inhibitor
develops (is acquired) after treatment with B-raf antagonist. In
other embodiments, the patient (e.g., the patient having B-raf
resistant cancer) has not been previously treated with a B-raf
antagonist.
[0124] In some embodiments, the patient is currently being treated
with B-raf antagonist, such as a B-raf inhibitor. In some
embodiments, the patient was previously treated with B-raf
antagonist. In some embodiments, the patient was not previously
treated with B-raf antagonist.
[0125] In some embodiments, the individual has colorectal cancer,
melanoma, lung cancer, ovarian cancer, breast cancer, pancreatic
cancer, hematological malignancy, bladder cancer, and/or renal cell
carcinoma. In some embodiments, the individual has non-small cell
lung cancer. The non-small cell lung cancer may be at early stage
or at late stage. In some embodiments, the individual has small
cell lung cancer. The small cell lung cancer may be at early stage
or at late stage. In some embodiments, the individual has renal
cell cancer. The renal cell cancer may be at early stage or at late
stage. In some embodiments, the individual has colorectal cancer.
The colorectal cancer may be at early stage or late stage. In some
embodiments, the individual has ovarian cancer. The ovarian cancer
may be at early stage or at late stage. In some embodiments, the
individual has breast cancer. The breast cancer may be at early
stage or at late stage. In some embodiments, the individual has
pancreatic cancer. The pancreatic cancer may be at early stage or
at late stage. In some embodiments, the individual has gastric
carcinoma. The gastric carcinoma may be at early stage or at late
stage. In some embodiments, the individual has bladder cancer. The
bladder cancer may be at early stage or at late stage. In some
embodiments, the individual has esophageal cancer. The esophageal
cancer may be at early stage or at late stage. In some embodiments,
the individual has mesothelioma. The mesothelioma may be at early
stage or at late stage. In some embodiments, the individual has
melanoma. The melanoma may be at early stage or at late stage. In
some embodiments, the individual has head and neck cancer. The head
and neck cancer may be at early stage or at late stage. In some
embodiments, the individual has thyroid cancer. The thyroid cancer
may be at early stage or at late stage. In some embodiments, the
individual has sarcoma. The sarcoma may be at early stage or late
stage. In some embodiments, the individual has prostate cancer. The
prostate cancer may be at early stage or at late stage. In some
embodiments, the individual has glioblastoma. The glioblastoma may
be at early stage or at late stage. In some embodiments, the
individual has cervical cancer. The cervical cancer may be at early
stage or at late stage. In some embodiments, the individual has
thymic carcinoma. The thymic carcinoma may be at early stage or at
late stage. In some embodiments, the individual has leukemia. The
leukemia may be at early stage or at late stage. In some
embodiments, the individual has lymphomas. The lymphoma may be at
early stage or at late stage. In some embodiments, the individual
has myelomas. The myelomas may be at early stage or at late stage.
In some embodiments, the individual has mycosis fungoides. The
mycosis fungoides may be at early stage or at late stage. In some
embodiments, the individual has merkel cell cancer. The merkel cell
cancer may be at early stage or at late stage. In some embodiments,
the individual has hematologic malignancies. The hematological
malignancies may be early stage or late stage. In some embodiments,
the individual is a human. In some embodiments, the cancer is
metastatic.
[0126] In some embodiments, the individual is a mammal, such as
domesticated animals (e.g., cows, sheep, cats, dogs, and horses),
primates (e.g., humans and non-human primates such as monkeys),
rabbits, and rodents (e.g., mice and rats). In some embodiments,
the individual treated is a human.
[0127] In another aspect, provided herein is a method of enhancing
immune function in an individual having cancer comprising
administering an effective amount of a PD-1 axis binding antagonist
and a MEK inhibitor.
[0128] In some embodiments, the CD8 T cells in the individual have
enhanced priming, activation, proliferation and/or cytolytic
activity relative to prior to the administration of the PD-1
pathway antagonist and the MEK inhibitor. In some embodiments, the
CD8 T cell priming is characterized by elevated CD44 expression
and/or enhanced cytolytic activity in CD8 T cells. In some
embodiments, the CD8 T cell activation is characterized by an
elevated frequency of .gamma.-IFN.sup.+ CD8 T cells. In some
embodiments, the CD8 T cell is an antigen-specific T-cell. In some
embodiments, the immune evasion by signaling through PD-L1 surface
expression is inhibited.
[0129] In some embodiments, the cancer cells in the individual have
elevated expression of MHC class I antigen expression relative to
prior to the administration of the PD-1 pathway antagonist and the
MEK inhibitor.
[0130] In some embodiments, the antigen presenting cells in the
individual have enhanced maturation and activation relative prior
to the administration of the PD-1 pathway antagonist and the MEK
inhibitor. In some embodiments, wherein the antigen presenting
cells are dendritic cells. In some embodiments, the maturation of
the antigen presenting cells is characterized by increased
frequency of CD83.sup.+ dendritic cells. In some embodiments, the
activation of the antigen presenting cells is characterized by
elevated expression of CD80 and CD86 on dendritic cells.
[0131] In some embodiments, the serum levels of cytokine IL-10
and/or chemokine IL-8, a human homolog of murine KC, in the
individual are reduced relative prior to the administration of the
anti-PD-L1 antibody and the MEK inhibitor.
[0132] In some embodiments, the cancer has elevated levels of
T-cell infiltration.
[0133] In some embodiments, the combination therapy of the
invention comprises administration of a PD-1 axis binding
antagonist and a MEK inhibitor. The PD-1 axis binding antagonist
and the MEK inhibitor may be administered in any suitable manner
known in the art. For example, The PD-1 axis binding antagonist and
the MEK inhibitor may be administered sequentially (at different
times) or concurrently (at the same time).
[0134] In some embodiments, the MEK inhibitor is administered
continuously. In some embodiments, the MEK inhibitor is
administered intermittently. In some embodiments, the MEK inhibitor
is administered before administration of the PD-1 axis binding
antagonist. In some embodiments, the MEK inhibitor is administered
simultaneously with administration of the PD-1 axis binding
antagonist. In some embodiments, the MEK inhibitor is administered
after administration of the PD-1 axis binding antagonist.
[0135] In some embodiments, provided is a method for treating or
delaying progression of cancer in an individual comprising
administering to the individual an effective amount of a PD-1 axis
binding antagonist and a MEK inhibitor, further comprising
administering an additional therapy. The additional therapy may be
radiation therapy, surgery (e.g., lumpectomy and a mastectomy),
chemotherapy, gene therapy, DNA therapy, viral therapy, RNA
therapy, immunotherapy, bone marrow transplantation, nanotherapy,
monoclonal antibody therapy, or a combination of the foregoing. The
additional therapy may be in the form of adjuvant or neoadjuvant
therapy. In some embodiments, the additional therapy is the
administration of small molecule enzymatic inhibitor or
anti-metastatic agent. In some embodiments, the additional therapy
is the administration of side-effect limiting agents (e.g., agents
intended to lessen the occurrence and/or severity of side effects
of treatment, such as anti-nausea agents, etc.). In some
embodiments, the additional therapy is radiation therapy. In some
embodiments, the additional therapy is surgery. In some
embodiments, the additional therapy is a combination of radiation
therapy and surgery. In some embodiments, the additional therapy is
gamma irradiation. In some embodiments, the additional therapy is
therapy targeting PI3K/AKT/mTOR pathway, HSP90 inhibitor, tubulin
inhibitor, apoptosis inhibitor, and/or chemopreventative agent. The
additional therapy may be one or more of the chemotherapeutic
agents described hereabove.
[0136] The PD-1 axis binding antagonist and the MEK inhibitor may
be administered by the same route of administration or by different
routes of administration. In some embodiments, the PD-1 axis
binding antagonist is administered intravenously, intramuscularly,
subcutaneously, topically, orally, transdermally,
intraperitoneally, intraorbitally, by implantation, by inhalation,
intrathecally, intraventricularly, or intranasally. In some
embodiments, the MEK inhibitor is administered intravenously,
intramuscularly, subcutaneously, topically, orally, transdermally,
intraperitoneally, intraorbitally, by implantation, by inhalation,
intrathecally, intraventricularly, or intranasally. An effective
amount of the PD-1 axis binding antagonist and the MEK inhibitor
may be administered for prevention or treatment of disease. The
appropriate dosage of the PD-1 axis binding antagonist and/or the
MEK inhibitor may be determined based on the type of disease to be
treated, the type of the PD-1 axis binding antagonist and the MEK
inhibitor, the severity and course of the disease, the clinical
condition of the individual, the individual's clinical history and
response to the treatment, and the discretion of the attending
physician.
[0137] Any of the PD-1 axis binding antagonists and the MEK
inhibitors known in the art or described below may be used in the
methods.
PD-1 Axis Binding Antagonists
[0138] Provided herein is a method for treating or delaying
progression of cancer in an individual comprising administering to
the individual an effective amount of a PD-1 axis binding
antagonist and a MEK inhibitor. For example, a PD-1 axis binding
antagonist includes a PD-1 binding antagonist, a PD-L1 binding
antagonist and a PD-L2 binding antagonist. Alternative names for
"PD-1" include CD279 and SLEB2. Alternative names for "PD-L1"
include B7-H1, B7-4, CD274, and B7-H. Alternative names for "PD-L2"
include B7-DC, Btdc, and CD273. In some embodiments, PD-1, PD-L1,
and PD-L2 are human PD-1, PD-L1 and PD-L2.
[0139] In some embodiments, the PD-1 binding antagonist is a
molecule that inhibits the binding of PD-1 to its ligand binding
partners. In a specific aspect the PD-1 ligand binding partners are
PD-L1 and/or PD-L2. In another embodiment, a PD-L1 binding
antagonist is a molecule that inhibits the binding of PD-L1 to its
binding partners. In a specific aspect, PD-L1 binding partners are
PD-1 and/or B7-1. In another embodiment, the PD-L2 binding
antagonist is a molecule that inhibits the binding of PD-L2 to its
binding partners. In a specific aspect, a PD-L2 binding partner is
PD-1. The antagonist may be an antibody, an antigen binding
fragment thereof, an immunoadhesin, a fusion protein, or
oligopeptide.
[0140] In some embodiment, the PD-1 binding antagonist is an
anti-PD-1 antibody (e.g., a human antibody, a humanized antibody,
or a chimeric antibody). In some embodiments, the anti-PD-1
antibody is selected from the group consisting of MDX-1106
(nivolumab, OPDIVO.RTM.), Merck 3745 (MK-3475, pembrolizumab,
KEYTRUDA.RTM.), CT-011 (pidilizumab), MEDI-0680 (AMP-514), PDR001,
REGN2810, BGB-108, and BGB-A317. In some embodiments, the PD-1
binding antagonist is an immunoadhesin (e.g., an immunoadhesin
comprising an extracellular or PD-1 binding portion of PD-L1 or
PD-L2 fused to a constant region (e.g., an Fc region of an
immunoglobulin sequence). In some embodiments, the PD-1 binding
antagonist is AMP-224. In some embodiments, the PD-L1 binding
antagonist is anti-PD-L1 antibody. In some embodiments, the
anti-PD-L1 binding antagonist is selected from the group consisting
of YW243.55.S70, MPDL3280A (atezolizumab), MEDI4736 (durvalumab),
MDX-1105, and MSB0010718C (avelumab). MDX-1105, also known as
BMS-936559, is an anti-PD-L1 antibody described in WO2007/005874.
Antibody YW243.55.S70 (heavy and light chain variable region
sequences shown in SEQ ID Nos: 20 and 21, respectively) is an
anti-PD-L1 described in WO 2010/077634 A1. MEDI4736 is an
anti-PD-L1 antibody described in WO2011/066389 and US2013/034559.
MDX-1106, also known as nivolumab, MDX-1106-04, ONO-4538,
BMS-936558, or OPDIVO.RTM., is an anti-PD-1 antibody described in
WO2006/121168. Merck 3745, also known as MK-3475, pembrolizumab,
lambrolizumab, KEYTRUDA.RTM., or SCH-900475, is an anti-PD-1
antibody described in WO2009/114335. CT-011, also known as hBAT or
hBAT-1, is an anti-PD-1 antibody described in WO2009/101611.
AMP-224, also known as B7-DCIg, is a PD-L2-Fc fusion soluble
receptor described in WO2010/027827 and WO2011/066342.
[0141] In some embodiments, the anti-PD-1 antibody is MDX-1106.
Alternative names for "MDX-1106" include MDX-1106-04, ONO-4538,
BMS-936558 or nivolumab. In some embodiments, the anti-PD-1
antibody is Nivolumab (CAS Registry Number: 946414-94-4). In a
still further embodiment, provided is an isolated anti-PD-1
antibody comprising a heavy chain variable region comprising the
heavy chain variable region amino acid sequence from SEQ ID NO:22
and/or a light chain variable region comprising the light chain
variable region amino acid sequence from SEQ ID NO:23. In a still
further embodiment, provided is an isolated anti-PD-1 antibody
comprising a heavy chain and/or a light chain sequence,
wherein:
[0142] (a) the heavy chain sequence has at least 85%, at least 90%,
at least 91%, at least 92%, at least 93%, at least 94%, at least
95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%
sequence identity to the heavy chain sequence:
TABLE-US-00002 (SEQ ID NO: 22) QVQLVES
GGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVA
VIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATN
DDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEP
VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVD
HKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVL
TVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL
YSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK,
or
[0143] (b) the light chain sequences has at least 85%, at least
90%, at least 91%, at least 92%, at least 93%, at least 94%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%
or 100% sequence identity to the light chain sequence:
TABLE-US-00003 (SEQ ID NO: 23)
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQ
GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV
DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG
LSSPVTKSFNRGEC.
[0144] Examples of anti-PD-L1 antibodies useful for the methods of
this invention, and methods for making thereof are described in PCT
patent application WO 2010/077634 A1, which is incorporated herein
by reference.
[0145] In some embodiments, the PD-1 axis binding antagonist is an
anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody
is capable of inhibiting binding between PD-L1 and PD-1 and/or
between PD-L1 and B7-1. In some embodiments, the anti-PD-L1
antibody is a monoclonal antibody. In some embodiments, the
anti-PD-L1 antibody is an antibody fragment selected from the group
consisting of Fab, Fab'-SH, Fv, scFv, and (Fab').sub.2 fragments.
In some embodiments, the anti-PD-L1 antibody is a humanized
antibody. In some embodiments, the anti-PD-L1 antibody is a human
antibody.
[0146] The anti-PD-L1 antibodies useful in this invention,
including compositions containing such antibodies, such as those
described in WO 2010/077634 A1, may be used in combination with a
MEK inhibitor to treat cancer. In some embodiments, the anti-PD-L1
antibody comprises a heavy chain variable region comprising the
amino acid sequence of SEQ ID NO:20 and a light chain variable
region comprising the amino acid sequence of SEQ ID NO:21.
[0147] In one embodiment, the anti-PD-L1 antibody contains a heavy
chain variable region polypeptide comprising an HVR-H1, HVR-H2 and
HVR-H3 sequence, wherein:
TABLE-US-00004 (SEQ ID NO: 1) (a) the HVR-H1 sequence is
GFTFSX.sub.1SWIH; (SEQ ID NO: 2) (b) the HVR-H2 sequence is
AWIX.sub.2PYGGSX.sub.3YYADSVKG; (SEQ ID NO: 3) (c) the HVR-H3
sequence is RHWPGGFDY ;
[0148] further wherein: X.sub.1 is D or G; X.sub.2 is S or L;
X.sub.3 is T or S.
[0149] In one specific aspect, X.sub.1 is D; X.sub.2 is S and
X.sub.3 is T. In another aspect, the polypeptide further comprises
variable region heavy chain framework sequences juxtaposed between
the HVRs according to the formula:
(HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4). In
yet another aspect, the framework sequences are derived from human
consensus framework sequences. In a further aspect, the framework
sequences are VH subgroup III consensus framework. In a still
further aspect, at least one of the framework sequences is the
following:
TABLE-US-00005 (SEQ ID NO: 4) HC-FR1 is EVQLVESGGGLVQPGGSLRLSCAAS
(SEQ ID NO: 5) HC-FR2 is WVRQAPGKGLEWV (SEQ ID NO: 6) HC-FR3 is
RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 7) HC-FR4 is
WGQGTLVTVSA.
[0150] In a still further aspect, the heavy chain polypeptide is
further combined with a variable region light chain comprising an
HVR-L1, HVR-L2 and HVR-L3, wherein:
TABLE-US-00006 (SEQ ID NO: 8) (a) the HVR-L1 sequence is
RASQX.sub.4X.sub.5X.sub.6TX.sub.7X.sub.8A; (SEQ ID NO: 9) (b) the
HVR-L2 sequence is SASX.sub.9LX.sub.10S,; (SEQ ID NO: 10) (c) the
HVR-L3 sequence is
QQX.sub.11X.sub.12X.sub.13X.sub.14PX.sub.15T;
[0151] further wherein: X.sub.4 is D or V; X.sub.5 is V or I;
X.sub.6 is S or N; X.sub.7 is A or F; X.sub.8 is V or L; X.sub.9 is
F or T; X.sub.10 is Y or A; X.sub.11 is Y, G, F, or S; X.sub.12 is
L, Y, F or W; X.sub.13 is Y, N, A, T, G, F or I; X.sub.14 is H, V,
P, T or I; X.sub.15 is A, W, R, P or T.
[0152] In a still further aspect, X.sub.4 is D; X.sub.5 is V;
X.sub.6 is S; X.sub.7 is A; X.sub.8 is V; X.sub.9 is F; X.sub.10 is
Y; Xu is Y; X.sub.12 is L; X.sub.13 is Y; X.sub.14 is H; X.sub.15
is A. In a still further aspect, the light chain further comprises
variable region light chain framework sequences juxtaposed between
the HVRs according to the formula:
(LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In
a still further aspect, the framework sequences are derived from
human consensus framework sequences. In a still further aspect, the
framework sequences are VL kappa I consensus framework. In a still
further aspect, at least one of the framework sequence is the
following:
TABLE-US-00007 (SEQ ID NO: 11) LC-FR1 is DIQMTQSPSSLSASVGDRVTITC
(SEQ ID NO: 12) LC-FR2 is WYQQKPGKAPKLLIY (SEQ ID NO: 13) LC-FR3 is
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 14) LC-FR4 is
FGQGTKVEIKR.
[0153] In another embodiment, provided is an isolated anti-PD-L1
antibody or antigen binding fragment comprising a heavy chain and a
light chain variable region sequence, wherein:
[0154] (a) the heavy chain comprises and HVR-H1, HVR-H2 and HVR-H3,
wherein further:
TABLE-US-00008 (SEQ ID NO: 1) (i) the HVR-H1 sequence is
GFTFSX.sub.1SWIH; (SEQ ID NO: 2) (ii) the HVR-H2 sequence is
AWIX.sub.2PYGGSX.sub.3YYADSVKG (SEQ ID NO: 3) (iii) the HVR-H3
sequence is RHWPGGFDY, and
[0155] (b) the light chain comprises and HVR-L1, HVR-L2 and HVR-L3,
wherein further:
TABLE-US-00009 (SEQ ID NO: 8) (i) the HVR-L1 sequence is
RASQX.sub.4X.sub.5X.sub.6TX.sub.7X.sub.8A (SEQ ID NO: 9) (ii) the
HVR-L2 sequence is SASX.sub.9LX.sub.10S; and (SEQ ID NO: 10) (iii)
the HVR-L3 sequence is
QQX.sub.11X.sub.12X.sub.13X.sub.14PX.sub.15T;
[0156] Further wherein: X.sub.1 is D or G; X.sub.2 is S or L;
X.sub.3 is T or S; X.sub.4 is D or V; X.sub.5 is V or I; X.sub.6 is
S or N; X.sub.7 is A or F; X.sub.8 is V or L; X.sub.9 is F or T;
X.sub.10 is Y or A; X.sub.11 is Y, G, F, or S; X.sub.12 is L, Y, F
or W; X.sub.13 is Y, N, A, T, G, F or I; X.sub.14 is H, V, P, T or
I; X.sub.15 is A, W, R, P or T.
[0157] In a specific aspect, X.sub.1 is D; X.sub.2 is S and X.sub.3
is T. In another aspect, X.sub.4 is D; X.sub.5 is V; X.sub.6 is S;
X.sub.7 is A; X.sub.8 is V; X.sub.9 is F; X.sub.10 is Y; X.sub.11
is Y; X.sub.12 is L; X.sub.13 is Y; X.sub.14 is H; X.sub.15 is A.
In yet another aspect, X.sub.1 is D; X.sub.2 is S and X.sub.3 is T,
X.sub.4 is D; X.sub.5 is V; X.sub.6 is S; X.sub.7 is A; X.sub.8 is
V; X.sub.9 is F; X.sub.10 is Y; X.sub.11 is Y; X.sub.12 is L;
X.sub.13 is Y; X.sub.14 is H and X.sub.15 is A.
[0158] In a further aspect, the heavy chain variable region
comprises one or more framework sequences juxtaposed between the
HVRs as:
(HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and
the light chain variable regions comprises one or more framework
sequences juxtaposed between the HVRs as:
(LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In
a still further aspect, the framework sequences are derived from
human consensus framework sequences. In a still further aspect, the
heavy chain framework sequences are derived from a Rabat subgroup
I, II, or III sequence. In a still further aspect, the heavy chain
framework sequence is a VH subgroup III consensus framework. In a
still further aspect, one or more of the heavy chain framework
sequences is the following:
TABLE-US-00010 (SEQ ID NO: 4) HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ
ID NO: 5) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 6) HC-FR3
RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 7) HC-FR4
WGQGTLVTVSA.
[0159] In a still further aspect, the light chain framework
sequences are derived from a Kabat kappa I, II, II or IV subgroup
sequence. In a still further aspect, the light chain framework
sequences are VL kappa I consensus framework. In a still further
aspect, one or more of the light chain framework sequences is the
following:
TABLE-US-00011 (SEQ ID NO: 11) LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ
ID NO: 12) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 13) LC-FR3
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 14) LC-FR4
FGQGTKVEIKR.
[0160] In a still further specific aspect, the antibody further
comprises a human or murine constant region. In a still further
aspect, the human constant region is selected from the group
consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further
specific aspect, the human constant region is IgG1. In a still
further aspect, the murine constant region is selected from the
group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still
further aspect, the murine constant region if IgG2A. In a still
further specific aspect, the antibody has reduced or minimal
effector function. In a still further specific aspect the minimal
effector function results from an "effector-less Fc mutation" or
aglycosylation. In still a further embodiment, the effector-less Fc
mutation is an N297A or D265A/N297A substitution in the constant
region.
[0161] In yet another embodiment, provided is an anti-PD-L1
antibody comprising a heavy chain and a light chain variable region
sequence, wherein: [0162] (a) the heavy chain further comprises and
HVR-H1, HVR-H2 and an HVR-H3 sequence having at least 85% sequence
identity to GFTFSDSWIH (SEQ ID NO: 15), AWISPYGGSTYYADSVKG (SEQ ID
NO: 16) and RHWPGGFDY (SEQ ID NOG), respectively, or [0163] (b) the
light chain further comprises an HVR-L1, HVR-L2 and an HVR-L3
sequence having at least 85% sequence identity to RASQDVSTAVA (SEQ
ID NO: 17), SASFLYS (SEQ ID NO: 18) and QQYLYHPAT (SEQ ID NO: 19),
respectively.
[0164] In a specific aspect, the sequence identity is 86%, 87%,
88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%.
In another aspect, the heavy chain variable region comprises one or
more framework sequences juxtaposed between the HVRs as:
(HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and
the light chain variable regions comprises one or more framework
sequences juxtaposed between the HVRs as:
(LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In
yet another aspect, the framework sequences are derived from human
consensus framework sequences. In a still further aspect, the heavy
chain framework sequences are derived from a Rabat subgroup I, II,
or III sequence. In a still further aspect, the heavy chain
framework sequence is a VH subgroup III consensus framework. In a
still further aspect, one or more of the heavy chain framework
sequences is the following:
TABLE-US-00012 (SEQ ID NO: 4) HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ
ID NO: 5) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 6) HC-FR3
RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 7) HC-FR4
WGQGTLVTVSA.
[0165] In a still further aspect, the light chain framework
sequences are derived from a Rabat kappa I, II, II or IV subgroup
sequence. In a still further aspect, the light chain framework
sequences are VL kappa I consensus framework. In a still further
aspect, one or more of the light chain framework sequences is the
following:
TABLE-US-00013 (SEQ ID NO: 11) LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ
ID NO: 12) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 13) LC-FR3
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 14) LC-FR4
FGQGTKVEIKR.
[0166] In a still further specific aspect, the antibody further
comprises a human or murine constant region. In a still further
aspect, the human constant region is selected from the group
consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further
specific aspect, the human constant region is IgG1. In a still
further aspect, the murine constant region is selected from the
group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still
further aspect, the murine constant region if IgG2A. In a still
further specific aspect, the antibody has reduced or minimal
effector function. In a still further specific aspect the minimal
effector function results from an "effector-less Fc mutation" or
aglycosylation. In still a further embodiment, the effector-less Fc
mutation is an N297A or D265A/N297A substitution in the constant
region.
[0167] In a still further embodiment, provided is an isolated
anti-PD-L1 antibody comprising a heavy chain and a light chain
variable region sequence, wherein:
[0168] (a) the heavy chain sequence has at least 85% sequence
identity to the heavy chain sequence:
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWIS
PYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSA
(SEQ ID NO:20), or
[0169] (b) the light chain sequence has at least 85% sequence
identity to the light chain sequence:
DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASF
LYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID
NO:21).
[0170] In a specific aspect, the sequence identity is 86%, 87%,
88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%.
In another aspect, the heavy chain variable region comprises one or
more framework sequences juxtaposed between the HVRs as:
(HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and
the light chain variable regions comprises one or more framework
sequences juxtaposed between the HVRs as:
(LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In
yet another aspect, the framework sequences are derived from human
consensus framework sequences. In a further aspect, the heavy chain
framework sequences are derived from a Rabat subgroup I, II, or III
sequence. In a still further aspect, the heavy chain framework
sequence is a VH subgroup III consensus framework. In a still
further aspect, one or more of the heavy chain framework sequences
is the following:
TABLE-US-00014 (SEQ ID NO: 4) HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ
ID NO: 5) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 6) HC-FR3
RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 7) HC-FR4
WGQGTLVTVSA.
[0171] In a still further aspect, the light chain framework
sequences are derived from a Rabat kappa I, II, II or IV subgroup
sequence. In a still further aspect, the light chain framework
sequences are VL kappa I consensus framework. In a still further
aspect, one or more of the light chain framework sequences is the
following:
TABLE-US-00015 (SEQ ID NO: 11) LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ
ID NO: 12) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 13) LC-FR3
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 14) LC-FR4
FGQGTKVEIKR.
[0172] In a still further specific aspect, the antibody further
comprises a human or murine constant region. In a still further
aspect, the human constant region is selected from the group
consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further
specific aspect, the human constant region is IgG1. In a still
further aspect, the murine constant region is selected from the
group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still
further aspect, the murine constant region if IgG2A. In a still
further specific aspect, the antibody has reduced or minimal
effector function. In a still further specific aspect, the minimal
effector function results from production in prokaryotic cells. In
a still further specific aspect the minimal effector function
results from an "effector-less Fc mutation" or aglycosylation. In
still a further embodiment, the effector-less Fc mutation is an
N297A or D265A/N297A substitution in the constant region.
[0173] In another further embodiment, provided is an isolated
anti-PD-L1 antibody comprising a heavy chain and a light chain
variable region sequence, wherein:
[0174] (a) the heavy chain sequence has at least 85% sequence
identity to the heavy chain sequence:
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWIS
PYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWG QGTLVTVSS
(SEQ ID NO:24), or
[0175] (b) the light chain sequence has at least 85% sequence
identity to the light chain sequence:
DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASF
LYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID
NO:21).
[0176] In a still further embodiment, provided is an isolated
anti-PDL1 antibody comprising a heavy chain and a light chain
variable region sequence, wherein:
[0177] (a) the heavy chain sequence has at least 85% sequence
identity to the heavy chain sequence:
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWI
SPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYW
GQGTLVTVSSASTK (SEQ ID NO:28), or
[0178] (b) the light chain sequence has at least 85% sequence
identity to the light chain sequence:
TABLE-US-00016 (SEQ ID NO: 21)
DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYS
ASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQ GTKVEIKR.
[0179] In a specific aspect, the sequence identity is 86%, 87%,
88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%.
In another aspect, the heavy chain variable region comprises one or
more framework sequences juxtaposed between the HVRs as:
(HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and
the light chain variable regions comprises one or more framework
sequences juxtaposed between the HVRs as:
(LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In
yet another aspect, the framework sequences are derived from human
consensus framework sequences. In a further aspect, the heavy chain
framework sequences are derived from a Rabat subgroup I, II, or III
sequence. In a still further aspect, the heavy chain framework
sequence is a VH subgroup III consensus framework. In a still
further aspect, one or more of the heavy chain framework sequences
is the following:
TABLE-US-00017 (SEQ ID NO: 4) HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ
ID NO: 5) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 6) HC-FR3
RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 25) HC-FR4
WGQGTLVTVSS.
[0180] In a still further aspect, the light chain framework
sequences are derived from a Rabat kappa I, II, II or IV subgroup
sequence. In a still further aspect, the light chain framework
sequences are VL kappa I consensus framework. In a still further
aspect, one or more of the light chain framework sequences is the
following:
TABLE-US-00018 (SEQ ID NO: 11) LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ
ID NO: 12) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 13) LC-FR3
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 14) LC-FR4
FGQGTKVEIKR.
[0181] In a still further specific aspect, the antibody further
comprises a human or murine constant region. In a still further
aspect, the human constant region is selected from the group
consisting of IgG1, IgG2, IgG2, IgG3, and IgG4. In a still further
specific aspect, the human constant region is IgG1. In a still
further aspect, the murine constant region is selected from the
group consisting of IgG1, IgG2A, IgG2B, and IgG3. In a still
further aspect, the murine constant region if IgG2A. In a still
further specific aspect, the antibody has reduced or minimal
effector function. In a still further specific aspect, the minimal
effector function results from production in prokaryotic cells. In
a still further specific aspect the minimal effector function
results from an "effector-less Fc mutation" or aglycosylation. In
still a further embodiment, the effector-less Fc mutation is an
N297A or D265A/N297A substitution in the constant region.
[0182] In yet another embodiment, the anti-PD-1 antibody is
MPDL3280A (atezolizumab). In a still further embodiment, provided
is an isolated anti-PD-1 antibody comprising a heavy chain variable
region comprising the heavy chain variable region amino acid
sequence from SEQ ID NO:24 and/or a light chain variable region
comprising the light chain variable region amino acid sequence from
SEQ ID NO:25. In a still further embodiment, provided is an
isolated anti-PD-1 antibody comprising a heavy chain and/or a light
chain sequence, wherein:
[0183] (a) the heavy chain sequence has at least 85%, at least 90%,
at least 91%, at least 92%, at least 93%, at least 94%, at least
95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%
sequence identity to the heavy chain sequence:
TABLE-US-00019 (SEQ ID NO: 26)
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAW
ISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRH
WPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAST
YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK,
or
[0184] (b) the light chain sequences has at least 85%, at least
90%, at least 91%, at least 92%, at least 93%, at least 94%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%
or 100% sequence identity to the light chain sequence:
TABLE-US-00020 (SEQ ID NO: 27)
DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYS
ASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQ
GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV
DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG
LSSPVTKSFNRGEC.
[0185] In a still further embodiment, the invention provides for
compositions comprising any of the above described anti-PD-L1
antibodies in combination with at least one
pharmaceutically-acceptable carrier.
[0186] In a still further embodiment, provided is an isolated
nucleic acid encoding a light chain or a heavy chain variable
region sequence of an anti-PD-L1 antibody, wherein: [0187] (a) the
heavy chain further comprises and HVR-H1, HVR-H2 and an HVR-H3
sequence having at least 85% sequence identity to GFTFSDSWIH (SEQ
ID NO: 15), AWISPYGGSTYYADSVKG (SEQ ID NO: 16) and RHWPGGFDY (SEQ
ID NOG), respectively, and [0188] (b) the light chain further
comprises an HVR-L1, HVR-L2 and an HVR-L3 sequence having at least
85% sequence identity to RASQDVSTAVA (SEQ ID NO: 17), SASFLYS (SEQ
ID NO: 18) and QQYLYHPAT (SEQ ID NO: 19), respectively.
[0189] In a specific aspect, the sequence identity is 86%, 87%,
88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%.
In aspect, the heavy chain variable region comprises one or more
framework sequences juxtaposed between the HVRs as:
(HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and
the light chain variable regions comprises one or more framework
sequences juxtaposed between the HVRs as:
(LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). In
yet another aspect, the framework sequences are derived from human
consensus framework sequences. In a further aspect, the heavy chain
framework sequences are derived from a Rabat subgroup I, II, or III
sequence. In a still further aspect, the heavy chain framework
sequence is a VH subgroup III consensus framework. In a still
further aspect, one or more of the heavy chain framework sequences
is the following:
TABLE-US-00021 (SEQ ID NO: 4) HC-FR1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ
ID NO: 5) HC-FR2 WVRQAPGKGLEWV (SEQ ID NO: 6) HC-FR3
RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 7) HC-FR4
WGQGTLVTVSA.
[0190] In a still further aspect, the light chain framework
sequences are derived from a Kabat kappa I, II, II or IV subgroup
sequence. In a still further aspect, the light chain framework
sequences are VL kappa I consensus framework. In a still further
aspect, one or more of the light chain framework sequences is the
following:
TABLE-US-00022 (SEQ ID NO: 11) LC-FR1 DIQMTQSPSSLSASVGDRVTITC (SEQ
ID NO: 12) LC-FR2 WYQQKPGKAPKLLIY (SEQ ID NO: 13) LC-FR3
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 14) LC-FR4
FGQGTKVEIKR.
[0191] In a still further specific aspect, the antibody described
herein (such as an anti-PD-1 antibody, an anti-PD-L1 antibody, or
an anti-PD-L2 antibody) further comprises a human or murine
constant region. In a still further aspect, the human constant
region is selected from the group consisting of IgG1, IgG2, IgG2,
IgG3, and IgG4. In a still further specific aspect, the human
constant region is IgG1. In a still further aspect, the murine
constant region is selected from the group consisting of IgG1,
IgG2A, IgG2B, and IgG3. In a still further aspect, the murine
constant region if IgG2A. In a still further specific aspect, the
antibody has reduced or minimal effector function. In a still
further specific aspect, the minimal effector function results from
production in prokaryotic cells. In a still further specific aspect
the minimal effector function results from an "effector-less Fc
mutation" or aglycosylation. In still a further aspect, the
effector-less Fc mutation is an N297A or D265A/N297A substitution
in the constant region.
[0192] In a still further aspect, provided herein are nucleic acids
encoding any of the antibodies described herein. In some
embodiments, the nucleic acid further comprises a vector suitable
for expression of the nucleic acid encoding any of the previously
described anti-PD-L1, anti-PD-1, or anti-PD-L2 antibodies. In a
still further specific aspect, the vector further comprises a host
cell suitable for expression of the nucleic acid. In a still
further specific aspect, the host cell is a eukaryotic cell or a
prokaryotic cell. In a still further specific aspect, the
eukaryotic cell is a mammalian cell, such as Chinese Hamster Ovary
(CHO).
[0193] The antibody or antigen binding fragment thereof, may be
made using methods known in the art, for example, by a process
comprising culturing a host cell containing nucleic acid encoding
any of the previously described anti-PD-L1, anti-PD-1, or
anti-PD-L2 antibodies or antigen-binding fragment in a form
suitable for expression, under conditions suitable to produce such
antibody or fragment, and recovering the antibody or fragment.
[0194] In a still further embodiment, the invention provides for a
composition comprising an anti-PD-L1, an anti-PD-1, or an
anti-PD-L2 antibody or antigen binding fragment thereof as provided
herein and at least one pharmaceutically acceptable carrier. In
some embodiments, the anti-PD-L1, anti-PD-1, or anti-PD-L2 antibody
or antigen binding fragment thereof administered to the individual
is a composition comprising one or more pharmaceutically acceptable
carrier. Any of the pharmaceutically acceptable carrier described
herein or known in the art may be used.
MEK Inhibitors
[0195] The invention provides methods for treating cancer or
slowing progression of cancer in an individual comprising
administering an effective amount of a PD-1 pathway antagonist and
a MEK inhibitor. Any known MEK inhibitors are intended, such as the
MEK inhibitor compounds described in PCT patent applications WO
03/077914 A1, WO 2005/121142 A1, WO 2007/044515 A1, WO 2008/024725
A1 and WO 2009/085983 A1, the content of which are incorporated
herein by reference. The MEK inhibitor administered may be in a
pharmaceutical composition or formulation. In some embodiments, the
pharmaceutical composition or formulation comprises one or more MEK
inhibitors described herein and a pharmaceutically acceptable
carrier or excipient.
[0196] In some embodiments, the MEK inhibitor is a competitive
inhibitor of MEK. In some embodiments, the MEK inhibitor is more
selective against an activating KRAS mutation. In some embodiments,
the MEK inhibitor is an allosteric inhibitor of MEK. In some
embodiments, the MEK inhibitor is more selective against an
activating B-raf mutation (e.g., B-raf V600E mutation). In some
embodiments, the MEK inhibitor binds and inhibits the activity of
MEK1 and/or MEK2 (such as human MEK1 and/or human MEK2).
[0197] In some embodiments, the MEK inhibitor is a compound
selected from the group consisting of GDC-0973 (also known as
"Cobimetinib" or "XL518"), G-38963, G02443714 (also known as
"AS703206"), G02442104 (also known as "GSK-1120212"), and G00039805
(also known as "AZD-6244"), or a pharmaceutically acceptable salt
or solvate thereof.
[0198] In some embodiments, the MEK inhibitor is a compound of
formula (I),
##STR00001##
or a pharmaceutically acceptable salt or solvate thereof, wherein
A, X, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, and
R.sup.7 are as defined in Group A, Group B, Group C, or Group
D:
Group A:
[0199] A is arylene optionally substituted with one, two, three or
four groups selected from R.sup.10, R.sup.12, R.sup.14, R.sup.16,
and R.sup.19 where R.sup.10, R.sup.12, R.sup.14 and R.sup.16 are
independently hydrogen, alkyl, alkenyl, alkynyl, halo, haloalkoxy,
hydroxy, alkoxy, amino, alkylamino, dialkylamino, haloalkyl,
--NHS(O).sub.2R.sup.8, --CN, --C(O)R.sup.8, --C(O)OR.sup.8,
--C(O)NR.sup.8R.sup.8' and --NR.sup.8C(O)R.sup.8' and where
R.sup.19 is hydrogen, alkyl, or alkenyl; [0200] X is alkyl, halo,
haloalkyl, or haloalkoxy; [0201] R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5 and R.sup.6 are independently hydrogen, halo,
nitro, --NR.sup.8R.sup.8', --OR.sup.8, --NHS(O).sub.2R.sup.8, --CN,
--S(O).sub.mR.sup.8, --S(O).sub.2NR.sup.8R.sup.8', --C(O)R.sup.8,
--C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8', --NR.sup.8C(O)OR.sup.8',
--NR.sup.8C(O)NR.sup.8'R.sup.8'', --NR.sup.8C(O)OR.sup.8',
--NR.sup.8C(O)R.sup.8', --CH.sub.2N(R.sup.25)(NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(N(R.sup.25a)(NO.sub.2)),
--CH.sub.2NR.sup.25C(.dbd.NH)(N(R.sup.25a)(CN)),
--CH.sub.2NR.sup.25C(.dbd.NH)(R.sup.25),
--CH.sub.2NR.sup.25C(NR.sup.25aR.sup.25b).dbd.CH(NO.sub.2), alkyl,
alkenyl, alkynyl, cycloalkyl, heteroaryl, or heterocycloalkyl;
where the alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl, and
heterocycloalkyl are independently optionally substituted with one,
two, three, four, five, six or seven groups independently selected
from halo, alkyl, haloalkyl, nitro, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted arylalkyl, optionally
substituted heteroaryl, --OR.sup.8, --NR.sup.8R.sup.8',
--NR.sup.8S(O).sub.2R.sup.9, --CN, --S(O).sub.mR.sup.9,
--C(O)R.sup.8, --C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8',
--NR.sup.8C(O)NR.sup.8'R.sup.8'', --NR.sup.8C(O)OR.sup.8' and
--NR.sup.8C(O)R.sup.8'; or one of R.sup.1 and R.sup.2 together with
the carbon to which they are attached, R.sup.3 and R.sup.4 together
with the carbon to which they are attached, and R.sup.5 and R.sup.6
together with the carbon to which they are attached form C(O) or
C(.dbd.NOH); [0202] m is 0, 1, or 2; [0203] R.sup.7 is hydrogen,
halo or alkyl; [0204] each R.sup.8, R.sup.8' and R.sup.8'' is
independently selected from hydrogen, hydroxy, optionally
substituted alkoxy, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,
heteroaryl, and heterocycloalkyl; where the alkyl, alkenyl,
alkynyl, aryl, cycloalkyl, heteroaryl, and heterocycloalkyl are
independently optionally substituted with one, two three, four, or
five groups independently selected from alkyl, halo, hydroxy,
hydroxyalkyl, optionally substituted alkoxy, alkoxyalkyl,
haloalkyl, carboxy, alkoxycarbonyl, alkenyloxycarbonyl, optionally
substituted cycloalkyl, optionally substituted
cycloalkyloxycarbonyl, optionally substituted aryl, optionally
substituted aryloxy, optionally substituted aryloxycarbonyl,
optionally substituted arylalkyl, optionally substituted
arylalkyloxy, optionally substituted arylalkyloxycarbonyl, nitro,
cyano, optionally substituted heterocycloalkyl, optionally
substituted heteroaryl, --S(O)R.sup.31 (where n is 0, 1, or 2 and
R.sup.31 is optionally substituted alkyl, optionally substituted
aryl, optionally substituted heterocycloalkyl, or optionally
substituted heteroaryl), --NR.sup.34SO.sub.2R.sup.34a (where
R.sup.34 is hydrogen or alkyl and R.sup.34a is alkyl, alkenyl,
cycloalkyl, aryl, heteroaryl, or heterocycloalkyl),
--SO.sub.2NR.sup.35R.sup.35a (where R.sup.35 is hydrogen or alkyl
and R.sup.35a is alkyl, alkenyl, cycloalkyl, aryl, heteroaryl, or
heterocycloalkyl), --NR.sup.32C(O)R.sup.32a (where R.sup.32 is
hydrogen or alkyl and R.sup.32a is alkyl, alkenyl, alkoxy, or
cycloalkyl), --NR.sup.30R.sup.30' (where R.sup.30 and R.sup.30' are
independently hydrogen, alkyl, or hydroxyalkyl), and
--C(O)NR.sup.33R.sup.33a (where R.sup.33 is hydrogen or alkyl and
R.sup.33a is alkyl, alkenyl, alkynyl, or cycloalkyl); and [0205]
each R.sup.9 is independently selected from alkyl, alkenyl,
alkynyl, aryl, cycloalkyl, heteroaryl, and heterocycloalkyl; where
the alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, and
heterocycloalkyl are independently optionally substituted with one,
two, three, four, or five groups selected from halo, hydroxy,
alkyl, haloalkyl, haloalkoxy, amino, alkylamino, and
dialkylamino;
Group B:
[0205] [0206] A is heteroarylene optionally substituted with one,
two, three, or four groups selected from R.sup.10, R.sup.12,
R.sup.14, R.sup.16 and R.sup.19 where R.sup.10, R.sup.12, R.sup.14
and R.sup.16 are independently hydrogen, alkyl, alkenyl, alkynyl,
halo, haloalkoxy, hydroxy, alkoxy, cyano, amino, alkylamino,
dialkylamino, haloalkyl, alkylsulfonylamino, alkylcarbonyl,
alkenylcarbonyl, alkoxycarbonyl, alkenyloxycarbonyl, aminocarbonyl,
alkylaminocarbonyl, dialkylaminocarbonyl, or alkylcarbonylamino;
where R.sup.19 is hydrogen, alkyl, or alkenyl; and where each alkyl
and alkenyl, either alone or as part of another group within
R.sup.10, R.sup.12, R.sup.14, R.sup.16, and R.sup.19, is
independently optionally substituted with halo, hydroxy, or alkoxy;
[0207] X is alkyl, halo, haloalkyl, or haloalkoxy; [0208] R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.3 and R.sup.6 are independently
hydrogen, halo, nitro, --NR.sup.8R.sup.8', --OR.sup.8,
--NHS(O).sub.2R.sup.8, --CN, --S(O).sub.mR.sup.8,
--S(O).sub.2NR.sup.8R.sup.8', --C(O)R.sup.8, --C(O)OR.sup.8,
--C(O)NR.sup.8R.sup.8', --NR.sup.8C(O)OR.sup.8',
--NR.sup.8C(O)NR.sup.8'R.sup.8'', --NR.sup.8C(O)OR.sup.8',
--NR.sup.8C(O)R.sup.8', --CH.sub.2N(R.sup.25)(NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(N(R.sup.25a)(NO.sub.2)),
--CH.sub.2NR.sup.25C(NH)(N(R.sup.25a)(CN)),
--CH.sub.2NR.sup.25C(.dbd.NH)(R.sup.25),
--CH.sub.2NR.sup.25C(NR.sup.25aR.sup.25b).dbd.CH(NO.sub.2), alkyl,
alkenyl, alkynyl, cycloalkyl, heteroaryl, or heterocycloalkyl,
where the alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl, and
heterocycloalkyl are independently optionally substituted with one,
two, three, four, five, six or seven groups independently selected
from halo, alkyl, haloalkyl, nitro, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted arylalkyl, optionally
substituted heteroaryl, --OR.sup.8, --NR.sup.8R.sup.8',
--NR.sup.8S(O).sub.2R.sup.9, --CN, --S(O).sub.mR.sup.9,
--C(O)R.sup.8, --C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8',
--NR.sup.8C(O)NR.sup.8'R.sup.8'', --NR.sup.8C(O)OR.sup.8' and
--NR.sup.8C(O)R.sup.8'; or one of R.sup.1 and R.sup.2 together with
the carbon to which they are attached, R.sup.3 and R.sup.4 together
with the carbon to which they are attached, and R.sup.5 and R.sup.6
together with the carbon to which they are attached form C(O) or
C(.dbd.NOH); [0209] m is 1 or 2; [0210] R.sup.7 is hydrogen, halo
or alkyl; and [0211] each R.sup.8, R.sup.8' and R.sup.8'' is
independently selected from hydrogen, hydroxy, optionally
substituted alkoxy, alkyl, haloalkyl, alkenyl, alkynyl, aryl,
cycloalkyl, heteroaryl, and heterocycloalkyl, where the alkyl,
alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, and
heterocycloalkyl are independently optionally substituted with one,
two three, four, or five groups independently selected from alkyl,
halo, hydroxy, hydroxyalkyl, optionally substituted alkoxy,
alkoxyalkyl, haloalkyl, carboxy, carboxy ester, nitro, cyano,
--S(O).sub.nR.sup.31 (where n is 0, 1, or 2 and R.sup.31 is
optionally substituted alkyl, optionally substituted aryl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, or optionally substituted heteroaryl),
--NR.sup.36S(O).sub.2R.sup.36a (where R.sup.36 is hydrogen, alkyl,
or alkenyl and R.sup.36a is alkyl, alkenyl, optionally substituted
aryl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, or optionally substituted heteroaryl),
--S(O).sub.2NR.sup.37R.sup.37a (where R.sup.37 is hydrogen, alkyl,
or alkenyl and R.sup.37a is alkyl, alkenyl, optionally substituted
aryl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, or optionally substituted heteroaryl), optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted arylalkyl,
optionally substituted aryloxy, optionally substituted
arylalkyloxy, optionally substituted heteroaryl, --NHC(O)R.sup.32
(where R.sup.32 is alkyl, alkenyl, alkoxy, or cycloalkyl) and
--NR.sup.30R.sup.30 (where R.sup.30 and R.sup.30' are independently
hydrogen, alkyl, or hydroxy alkyl), and --C(O)NHR.sup.33 (where
R.sup.33 is alkyl, alkenyl, alkynyl, or cycloalkyl);
Group C:
[0211] [0212] A is
[0212] ##STR00002## [0213] where R.sup.10 is hydrogen, alkyl,
alkenyl, alkynyl, halo, haloalkoxy, hydroxy, alkoxy, amino,
alkylamino, dialkylamino, haloalkyl, --NHS(O).sub.2R.sup.8, --CN,
--C(O)R.sup.8, --C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8 and
--NR.sup.8C(O)R.sup.8'; [0214] R.sup.10a is hydrogen, alkyl, or
alkenyl; [0215] Y.sup.1 is .dbd.CH-- or .dbd.N--; [0216] X is
alkyl, halo, haloalkyl, or haloalkoxy; [0217] R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are independently hydrogen,
halo, nitro, --NR.sup.8R.sup.8', --OR.sup.8, --NHS(O).sub.2R.sup.8,
--CN, --S(O).sub.mR.sup.8, --S(O).sub.2NR.sup.8R.sup.8',
--C(O)R.sup.8, --C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8',
--NR.sup.8C(O)OR.sup.8', --NR.sup.8C(O)NR.sup.8'R.sup.8'',
--NR.sup.8C(O)OR.sup.8', --NR.sup.8C(O)R.sup.8',
--CH.sub.2N(R.sup.25)NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(N(R.sup.25a)(NO.sub.2)),
--CH.sub.2NR.sup.25C(.dbd.NH)(N(R.sup.25a)(CN)),
--CH.sub.2NR.sup.25C(.dbd.NH)(R.sup.25),
--CH.sub.2NR.sup.25C(NR.sup.25aR.sup.25b).dbd.CH(NO.sub.2), alkyl,
alkenyl, alkynyl, cycloalkyl, heteroaryl, or heterocycloalkyl,
where the alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl, and
heterocycloalkyl are independently optionally substituted with one,
two, three, four, five, six or seven groups independently selected
from halo, alkyl, haloalkyl, nitro, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted arylalkyl, optionally
substituted heteroaryl, --OR.sup.8, --NR.sup.8R.sup.8',
--NR.sup.8S(O).sub.2R.sup.9, --CN, --S(O).sub.mR.sup.9,
--C(O)R.sup.8, --C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8',
--NR.sup.8C(O)NR.sup.8'R.sup.8'', --NR.sup.8C(O)OR.sup.8' and
--NR.sup.8C(O)R.sup.8'; or one of R.sup.1 and R.sup.2 together with
the carbon to which they are attached, R.sup.3 and R.sup.4 together
with the carbon to which they are attached, and R.sup.5 and R.sup.6
together with the carbon to which they are attached form C(O) or
C(NOH); [0218] m is 1 or 2; [0219] R.sup.7 is hydrogen, halo or
alkyl; and [0220] each R.sup.8, R.sup.8' and R.sup.8'' is
independently selected from hydrogen, hydroxy, optionally
substituted alkoxy, alkyl, haloalkyl, alkenyl, alkynyl, aryl,
cycloalkyl, heteroaryl, and heterocycloalkyl, where the alkyl,
alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, and
heterocycloalkyl are independently optionally substituted with one,
two three, four, or five groups independently selected from alkyl,
halo, hydroxy, hydroxyalkyl, optionally substituted alkoxy,
alkoxyalkyl, haloalkyl, carboxy, carboxy ester, nitro, cyano,
--S(O).sub.nR.sup.31 (where n is 0, 1, or 2 and R.sup.31 is
optionally substituted alkyl, optionally substituted aryl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, or optionally substituted heteroaryl),
--NR.sup.36S(O).sub.2R.sup.36a (where R.sup.36 is hydrogen, alkyl,
or alkenyl and R.sup.36a is alkyl, alkenyl, optionally substituted
aryl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, or optionally substituted heteroaryl),
--S(O).sub.2NR.sup.37R.sup.37a (where R.sup.37 is hydrogen, alkyl,
or alkenyl and R.sup.37a is alkyl, alkenyl, optionally substituted
aryl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, or optionally substituted heteroaryl), optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted arylalkyl,
optionally substituted aryloxy, optionally substituted
arylalkyloxy, optionally substituted heteroaryl, --NHC(O)R.sup.32
(where R.sup.32 is alkyl, alkenyl, alkoxy, or cycloalkyl) and
--NR.sup.30R.sup.30 (where R.sup.30 and R.sup.30' are independently
hydrogen, alkyl, or hydroxyalkyl), and --C(O)NHR.sup.33 (where
R.sup.33 is alkyl, alkenyl, alkynyl, or cycloalkyl); or
Group D:
[0220] [0221] A is
[0221] ##STR00003## [0222] R.sup.40 and R.sup.40a are independently
hydrogen or alkyl; [0223] X is alkyl, halo, haloalkyl, or
haloalkoxy; [0224] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5 and
R.sup.6 are independently hydrogen, halo, nitro,
--NR.sup.8R.sup.8', --OR.sup.8, --NHS(O).sub.2R.sup.8, --CN,
--S(O).sub.mR.sup.8, --S(O).sub.2NR.sup.8R.sup.8', --C(O)R.sup.8,
--C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8', --NR.sup.8C(O)OR.sup.8',
--NR.sup.8C(O)NR.sup.8'R.sup.8'', --NR.sup.8C(O)OR.sup.8',
--NR.sup.8C(O)R.sup.8', --CH.sub.2N(R.sup.25)(NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(NR.sup.25aR.sup.25b),
--CH.sub.2NR.sup.25C(.dbd.NH)(N(R.sup.25a)(NO.sub.2)),
--CH.sub.2NR.sup.25C(.dbd.NH)(N(R.sup.25a)(CN)),
--CH.sub.2NR.sup.25C(.dbd.NH)(R.sup.25),
--CH.sub.2NR.sup.25C(NR.sup.25aR.sup.25b).dbd.CH(NO.sub.2), alkyl,
alkenyl, alkynyl, cycloalkyl, heteroaryl, or heterocycloalkyl,
where the alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl, and
heterocycloalkyl are independently optionally substituted with one,
two, three, four, five, six or seven groups independently selected
from halo, alkyl, haloalkyl, nitro, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted arylalkyl, optionally
substituted heteroaryl, --OR.sup.8, --NR.sup.8R.sup.8',
--NR.sup.8S(O).sub.2R.sup.9, --CN, --S(O).sub.mR.sup.9,
--C(O)R.sup.8, --C(O)OR.sup.8, --C(O)NR.sup.8R.sup.8',
--NR.sup.8C(O)NR.sup.8'R.sup.8'', --NR.sup.8C(O)OR.sup.8' and
--NR.sup.8C(O)R.sup.8'; or one of R.sup.1 and R.sup.2 together with
the carbon to which they are attached, R.sup.3 and R.sup.4 together
with the carbon to which they are attached, and R.sup.5 and R.sup.6
together with the carbon to which they are attached form C(O) or
C(NOH); [0225] m is 1 or 2; [0226] R.sup.7 is hydrogen, halo or
alkyl; and [0227] R.sup.8, R.sup.8' and R.sup.8'' are independently
selected from hydrogen, hydroxy, optionally substituted alkoxy,
alkyl, haloalkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl,
and heterocycloalkyl, where the alkyl, alkenyl, alkynyl, aryl,
cycloalkyl, heteroaryl, and heterocycloalkyl are independently
optionally substituted with one, two three, four, or five groups
independently selected from alkyl, halo, hydroxy, hydroxyalkyl,
optionally substituted alkoxy, alkoxyalkyl, haloalkyl, carboxy,
carboxy ester, nitro, cyano, --S(O).sub.nR.sup.31 (where n is 0, 1,
or 2 and R.sup.31 is optionally substituted alkyl, optionally
substituted aryl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, or optionally substituted
heteroaryl), --NR.sup.36S(O).sub.2R.sup.36a (where R.sup.36 is
hydrogen, alkyl, or alkenyl and R.sup.36a is alkyl, alkenyl,
optionally substituted aryl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, or optionally substituted
heteroaryl), --S(O).sub.2NR.sup.37R.sup.37a (where R.sup.37 is
hydrogen, alkyl, or alkenyl and R.sup.37a is alkyl, alkenyl,
optionally substituted aryl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, or optionally substituted
heteroaryl), optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted arylalkyl, optionally substituted aryloxy,
optionally substituted arylalkyloxy, optionally substituted
heteroaryl, --NHC(O)R.sup.32 (where R.sup.32 is alkyl, alkenyl,
alkoxy, or cycloalkyl) and --NR.sup.30R.sup.30 (where R.sup.30 and
R.sup.30' are independently hydrogen, alkyl, or hydroxy alkyl), and
--C(O)NHR.sup.33 (where R.sup.33 is alkyl, alkenyl, alkynyl, or
cycloalkyl).
[0228] In some variations, the MEK inhibitor compound of the
formula (I) is a compound of the Group A, having the formula I(a)
or I(b):
##STR00004##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined for the formula (I), Group A, or as
defined in WO 2007/044515 A1, incorporated herein by reference.
[0229] In some variations, the MEK inhibitor compound of the
formula (I) is a compound of the Group B, having the formula I(c),
I(d), I(e), I(f), I(g), I(h), I(i), I(j), I(k), I(m), I(n), I(o),
I(p), I(q), I(r), I(s), I(u), I(v), I(w), I(x), I(cc) or I(dd):
##STR00005## ##STR00006## ##STR00007## ##STR00008## ##STR00009##
##STR00010##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined for the formula (I), Group B, or as
defined in WO 2007/044515 A1, incorporated herein by reference.
[0230] In some variations, the MEK inhibitor compound of the
formula (I) is a compound of the Group C, having the formula I(y)
or I(z):
##STR00011##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined for the formula (I), Group C, or as
defined in WO 2007/044515 A1, incorporated herein by reference.
[0231] In some variations, the MEK inhibitor compound of the
formula (I) is a compound of the Group D, having the formula I(aa)
or I(bb):
##STR00012##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined for the formula (I), Group D, or as
defined in WO 2007/044515 A1, incorporated herein by reference.
[0232] In some embodiments, the MEK inhibitor compound of the
formula (I) is a compound selected from the compound Nos. 1-362 as
listed in WO 2007/044515 A1, Table 1 on pages 71-144 (herein
collectively referred to as the Formula I Species), or a
pharmaceutically acceptable salt or solvate thereof.
[0233] Also embraced are any variations of formula (I) as described
in WO 2007/044515 A1, which is incorporated herein by reference.
Compounds of the formula (I) or any variations thereof can be
synthesized using methods known in the art, for example, the
synthetic methods described in WO 2007/044515 A1, incorporated
herein by reference.
[0234] Unless defined otherwise herein, the terms used in
describing compounds of the formula (I) should be understood to
have the same meaning as defined in WO 2007/044515 A1.
[0235] In some embodiments, the MEK inhibitor is a compound of
formula (II):
##STR00013##
or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0236] Z.sup.1 is CR.sup.1 or N;
[0237] Z.sup.2 is CR.sup.2 or N;
[0238] Z.sup.3 is CR.sup.3 or N;
[0239] Z.sup.4 is CR.sup.4 or N;
[0240] where one or two of Z.sup.1, Z.sup.2, Z.sup.3, and Z.sup.4
are N;
[0241] R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently
selected from H, halo, CN, CF.sub.3, --OCF.sub.3, --NO.sub.2,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y)R.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y)OR.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y)NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nNR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nOR.sup.11,
--(CR.sup.14R.sup.15).sub.nSR.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.12C(.dbd.Y)R.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.12C(.dbd.Y)OR.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.13C(.dbd.Y)NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nNR.sup.12SO.sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y)R.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y)OR.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y)NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nOS(O).sub.2(OR.sup.11),
--(CR.sup.14R.sup.15).sub.nOP(.dbd.Y)(OR.sup.11)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.nOP(OR.sup.11)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.nS(O)R.sup.11,
--(CR.sup.14R.sup.15).sub.nS(O).sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.n S(O).sub.2NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nS(O)(OR.sup.11),
--(CR.sup.14R.sup.15).sub.nS(O).sub.2(OR.sup.11),
--(CR.sup.14R.sup.15).sub.nSC(.dbd.Y)R.sup.11,
--(CR.sup.14R.sup.15).sub.nSC(.dbd.Y)OR.sup.11,
--(CR.sup.14R.sup.15).sub.nSC(.dbd.Y)NR.sup.11R.sup.12,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl;
[0242] W is
##STR00014##
[0243] R.sup.5 and R.sup.6 are independently selected from H or
C.sub.1-C.sub.12 alkyl;
[0244] X.sup.1 is selected from R.sup.11, --OR.sup.11,
--NR.sup.11R.sup.12, --S(O)R.sup.11, and --S(O).sub.2R.sup.11; when
X.sup.1 is R.sup.11 or --OR.sup.11, R.sup.11 or --OR.sup.11 of
X.sup.1 and --R.sup.5 are optionally taken together with the
nitrogen atom to which they are attached to form a 4-7 membered
saturated or unsaturated ring having 0-2 additional heteroatoms
selected from O, S and N, wherein said ring is optionally
substituted with one or more groups selected from halo, CN,
CF.sub.3, --OCF.sub.3, --NO.sub.2, oxo, --Si(C.sub.1-C.sub.6
alkyl), --(CR.sup.19R.sup.20).sub.nC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.n C(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOR.sup.16,
--(CR.sup.19R.sup.20).sub.n--SR.sup.16, --(CR.sup.19R.sup.20).sub.n
NR.sup.16C(.dbd.Y')R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.16C(.dbd.Y')OR.sup.17,
--(CR.sup.19R.sup.20).sub.n NR.sup.18C(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.17SO.sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOS(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.nOP(.dbd.Y')(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nOP(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nS(O)R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nS(O)(OR.sup.16),
--(CR.sup.19R.sup.20).sub.n S(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')NR.sup.16R.sup.17, and
R.sup.21;
[0245] X.sup.2 is selected from carbocyclyl, heterocyclyl, aryl,
and heteroaryl;
[0246] R.sup.11, R.sup.12 and R.sup.13 are independently H,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl,
[0247] or R.sup.11 and R.sup.12 together with the nitrogen to which
they are attached form a 3-8 membered saturated, unsaturated or
aromatic ring having 0-2 heteroatoms selected from O, S and N,
wherein said ring is optionally substituted with one or more groups
selected from halo, CN, CF.sub.3, --OCF.sub.3, --NO.sub.2,
C.sub.1-C.sub.6 alkyl, --OH, --SH, --O(C.sub.1-C.sub.6 alkyl),
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0248] R.sup.14 and R.sup.15 are independently selected from H,
C.sub.1-C.sub.12 alkyl, aryl, carbocyclyl, heterocyclyl, and
heteroaryl;
[0249] m and n are independently selected from 0, 1, 2, 3, 4, 5, or
6;
[0250] Y is independently O, NR.sup.11, or S;
[0251] wherein each said alkyl, alkenyl, alkynyl, carbocyclyl,
heterocyclyl, aryl and heteroaryl of R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5, R.sup.6, X.sup.1, X.sup.2, R.sup.11, R.sup.12,
R.sup.13, R.sup.14, and R.sup.15 is independently optionally
substituted with one or more groups independently selected from
halo, CN, CF.sub.3, --OCF.sub.3, --NO.sub.2, oxo,
--Si(C.sub.1-C.sub.6 alkyl),
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.n C(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOR.sup.16,
--(CR.sup.19R.sup.20).sub.n--SR.sup.16, --(CR.sup.19R.sup.20).sub.n
NR.sup.16C(.dbd.Y')R.sup.17, --(CR.sup.19R.sup.20).sub.n
NR.sup.16C(.dbd.Y')OR.sup.17, --(CR.sup.19R.sup.20).sub.n
NR.sup.18C(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.17SO.sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOS(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.nOP(.dbd.Y')(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nOP(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nS(O)R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nS(O)(OR.sup.16),
--(CR.sup.19R.sup.20).sub.n S(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')NR.sup.16R.sup.17, and
R.sup.21;
[0252] each R.sup.16, R.sup.17 and R.sup.18 is independently H,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, wherein
said alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or
heteroaryl is optionally substituted with one or more groups
selected from halo, oxo, CN, --OCF.sub.3, CF.sub.3, --NO.sub.2,
C.sub.1-C.sub.6 alkyl, --OH, --SH, --O(C.sub.1-C.sub.6 alkyl),
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0253] or R.sup.16 and R.sup.17 together with the nitrogen to which
they are attached form a 3-8 membered saturated, unsaturated or
aromatic ring having 0-2 heteroatoms selected from O, S and N,
wherein said ring is optionally substituted with one or more groups
selected from halo, CN, --OCF.sub.3, CF.sub.3, --NO.sub.2,
C.sub.1-C.sub.6 alkyl, --OH, --SH, --O(C.sub.1-C.sub.6 alkyl),
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0254] R.sup.19 and R.sup.20 are independently selected from H,
C.sub.1-C.sub.12 alkyl, --(CH.sub.2).sub.n-aryl,
--(CH.sub.2).sub.n-carbocyclyl, --(CH.sub.2).sub.n-heterocyclyl,
and --(CH.sub.2).sub.n-heteroaryl;
[0255] R.sup.21 is C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl,
C.sub.2-C.sub.8 alkynyl, carbocyclyl, heterocyclyl, aryl, or
heteroaryl, wherein each member of R.sup.21 is optionally
substituted with one or more groups selected from halo, CN,
--OCF.sub.3, CF.sub.3, --NO.sub.2, C.sub.1-C.sub.6 alkyl, --OH,
--SH, --O(C.sub.1-C.sub.6 alkyl), --S(C.sub.1-C.sub.6 alkyl),
--NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0256] each Y' is independently O, NR.sup.22, or S; and
[0257] R.sup.22 is H or C.sub.1-C.sub.12 alkyl.
[0258] In some variations, the MEK inhibitor compound of the
formula (II) is a compound of the formula (II-1-a), (II-1-b),
(II-1-c), (II-1-d), (II-1-e), (II-1-f), (II-1-g), (II-1-h),
(II-1-i), (II-2-a), (II-2-b), (II-2-c), (II-2-d), (II-2-e),
(II-2-f), (II-2-g), (II-2-h), (II-2-i), (II-3-a), (II-3-b),
(II-3-c), (II-3-d), (II-3-e), (II-3-f), (II-3-g), (II-3-h), or
(II-3-i):
##STR00015## ##STR00016## ##STR00017## ##STR00018## ##STR00019##
##STR00020##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined for the formula (II) or as defined in
WO 2008/024725 A1, incorporated herein by reference.
[0259] In some embodiments, the MEK inhibitor compound of the
formula (II) is a compound selected from the compounds of Examples
5-18, 20-102, 105-109, 111-118, 120-133, 136-149 and 151-160 in WO
2008/024725 A1 (herein collectively referred to as the Formula II
Species), or a pharmaceutically acceptable salt or solvate thereof.
These compounds exhibited an IC.sub.50 of less than 10 .mu.M in the
assay described either in Example 8a or 8b (MEK activity assays).
Most of these compounds exhibited an IC.sub.50 of less than 5
.mu.M. See page 62 in WO 2008/024725 A1.
[0260] Also embraced are MEK inhibitor compounds (and/or solvates
and salts thereof) described in WO 2008/024725 A1, which is
incorporated herein by reference, for example, aza-benzofuran
compounds of the formula (II) (designated as formula I in WO
2008/024725 A1, e.g., on page 3) and variations thereof as
described in WO 2008/024725 A1. Compounds of formula (II) can be
synthesized using methods known in the art, for example, the
synthetic methods described in WO 2008/024725 A1, incorporated
herein by reference.
[0261] In some embodiments, the MEK inhibitor is a compound of
formula (III):
##STR00021##
or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0262] Z.sup.1 is CR.sup.1 or N;
[0263] R.sup.1 is H, C.sub.1-C.sub.3 alkyl, halo, CF.sub.3,
CHF.sub.2, CN, OR.sup.A or NR.sup.AR.sup.A;
[0264] R.sup.1 is H, C.sub.1-C.sub.3 alkyl, halo, CF.sub.3,
CHF.sub.2, CN, OR.sup.A, or NR.sup.AR.sup.A;
[0265] wherein each R.sup.A is independently H or C.sub.1-C.sub.3
alkyl;
[0266] Z.sup.2 is CR.sup.2 or N;
[0267] Z.sup.3 is CR.sup.3 or N; provided that only one of Z.sup.1,
Z.sup.2 and Z.sup.3 can be N at the same time;
[0268] R.sup.2 and R.sup.3 are independently selected from H, halo,
CN, CF.sub.3, --OCF.sub.3, --NO.sub.2,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nNR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nOR.sup.11,
--(CR.sup.14R.sup.15).sub.nSR.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.12C(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.12C(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.13C(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nNR.sup.12SO.sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nOS(O).sub.2(OR.sup.11),
--(CR.sup.14R.sup.15).sub.nOP(.dbd.Y')(OR.sup.11)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.nOP(OR.sup.u)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.nS(O)R.sup.11,
--(CR.sup.14R.sup.15).sub.nS(O).sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.n S(O).sub.2NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nS(O)(OR.sup.11),
--(CR.sup.14R.sup.15).sub.nS(O).sub.2(OR.sup.11),
--(CR.sup.14R.sup.15).sub.n SC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nSC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nSC(.dbd.Y')NR.sup.11R.sup.12,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl;
[0269] R.sup.4 is H, C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.4
carbocyclyl;
[0270] Y is W--C(O)-- or W';
[0271] W is
##STR00022##
[0272] R.sup.5 is H or C.sub.1-C.sub.12 alkyl;
[0273] X.sup.1 is selected from R.sup.11 and --OR.sup.11; when
X.sup.1 is R.sup.11, X.sup.1 is optionally taken together with
R.sup.5 and the nitrogen atom to which they are bound to form a 4-7
membered saturated or unsaturated ring having 0-2 additional
heteroatoms selected from O, S and N, wherein said ring is
optionally substituted with one or more groups selected from halo,
CN, CF.sub.3, --OCF.sub.3, --NO.sub.2, oxo,
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOR.sup.16,
--(CR.sup.19R.sup.20).sub.n--SR.sup.16, --(CR.sup.19R.sup.20).sub.n
NR.sup.16C(.dbd.Y')R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.16C(.dbd.Y')OR.sup.17,
--(CR.sup.19R.sup.20).sub.n NR.sup.18C(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.17SO.sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOS(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.nOP(.dbd.Y')(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nOP(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nS(O)R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nS(O)(OR.sup.16),
--(CR.sup.19R.sup.20).sub.n S(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')NR.sup.16R.sup.17, and
R.sup.21;
[0274] each R.sup.11' is independently H, C.sub.1-C.sub.12 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, carbocyclyl,
heterocyclyl, aryl, or heteroaryl;
[0275] R.sup.11, R.sup.12 and R.sup.13 are independently H,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl,
[0276] or R.sup.11 and R.sup.12 together with the nitrogen to which
they are attached form a 3-8 membered saturated, unsaturated or
aromatic ring having 0-2 heteroatoms selected from O, S and N,
wherein said ring is optionally substituted with one or more groups
selected from halo, CN, CF.sub.3, --OCF.sub.3, --NO.sub.2,
C.sub.1-C.sub.6 alkyl, --OH, --SH, --O(C.sub.1-C.sub.6 alkyl),
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0277] R.sup.14 and R.sup.15 are independently selected from H,
C.sub.1-C.sub.12 alkyl, aryl, carbocyclyl, heterocyclyl, and
heteroaryl;
[0278] W is
##STR00023##
wherein
##STR00024##
is
##STR00025##
[0279] each X.sup.2 is independently O, S, or NR.sup.9;
[0280] each R.sup.7 is independently selected from H, halo, CN,
CF.sub.3, --OCF.sub.3, --NO.sub.2,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nNR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nOR.sup.11,
--(CR.sup.14R.sup.15).sub.nSR.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.12C(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.12C(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nNR.sup.13C(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nNR.sup.12SO.sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nOC(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nOS(O).sub.2(OR.sup.11),
--(CR.sup.14R.sup.15).sub.nOP(.dbd.Y')(OR.sup.11)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.nOP(OR.sup.11)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.nS(O)R.sup.11,
--(CR.sup.14R.sup.15).sub.nS(O).sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.n S(O).sub.2NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.nS(O)(OR.sup.11),
--(CR.sup.14R.sup.15).sub.nS(O).sub.2(OR.sup.11),
--(CR.sup.14R.sup.15).sub.n SC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nSC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nSC(.dbd.Y')NR.sup.11R.sup.12,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl;
[0281] each R.sup.8 is independently selected from C.sub.1-C.sub.12
alkyl, aryl, carbocyclyl, heterocyclyl, and heteroaryl;
[0282] R.sup.9 is selected from H,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.nC(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.qNR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.qOR.sup.11,
--(CR.sup.14R.sup.15).sub.qSR.sup.11,
--(CR.sup.14R.sup.15).sub.qNR.sup.12C(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.qNR.sup.12C(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.qNR.sup.13C(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.qNR.sup.12SO.sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.qOC(.dbd.Y')R.sup.11,
--(CR.sup.14R.sup.15).sub.qOC(.dbd.Y')OR.sup.11,
--(CR.sup.14R.sup.15).sub.qOC(.dbd.Y')NR.sup.11R.sup.12,
--(CR.sup.14R.sup.15).sub.qOS(O).sub.2(OR.sup.11),
--(CR.sup.14R.sup.15).sub.qOP(.dbd.Y')(OR.sup.11)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.qOP(OR.sup.11)(OR.sup.12),
--(CR.sup.14R.sup.15).sub.nS(O)R.sup.11,
--(CR.sup.14R.sup.15).sub.nS(O).sub.2R.sup.11,
--(CR.sup.14R.sup.15).sub.n S(O).sub.2NR.sup.11R.sup.12,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl;
[0283] R.sup.10 is H, C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.4
carbocyclyl;
[0284] X.sup.4 is
##STR00026##
[0285] R.sup.6 is H, halo, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.8
alkenyl, C.sub.2-C.sub.8 alkynyl, carbocyclyl, heteroaryl,
heterocyclyl, --OCF.sub.3, --NO.sub.2, --Si(C.sub.1-C.sub.6 alkyl),
--(CR.sup.19R.sup.20).sub.nNR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOR.sup.16, or
--(CR.sup.19R.sup.20).sub.n--SR.sup.16;
[0286] R.sup.6' is H, halo, C.sub.1-C.sub.6 alkyl, carbocyclyl,
CF.sub.3, --OCF.sub.3, --NO.sub.2, --Si(C.sub.1-C.sub.6 alkyl),
--(CR.sup.19R.sup.20).sub.nNR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOR.sup.16,
--(CR.sup.19R.sup.20).sub.n--SR.sup.16, C.sub.2-C.sub.8 alkenyl,
C.sub.2-C.sub.8 alkynyl, heterocyclyl, aryl, or heteroaryl;
[0287] p is 0, 1, 2 or 3;
[0288] n is 0, 1, 2 or 3;
[0289] q is 2 or 3;
[0290] wherein each said alkyl, alkenyl, alkynyl, carbocyclyl,
heterocyclyl, aryl and heteroaryl of R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5, R.sup.6, R.sup.6', R.sup.7, R.sup.8, R.sup.9,
R.sup.10, R.sup.11, R.sup.11', R.sup.12, R.sup.13, R.sup.14,
R.sup.15 and R.sup.A is independently optionally substituted with
one or more groups independently selected from halo, CN, CF.sub.3,
--OCF.sub.3, --NO.sub.2, oxo, --Si(C.sub.1-C.sub.6 alkyl),
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.n C(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOR.sup.16,
--(CR.sup.19R.sup.20).sub.nSR.sup.16,
--(CR.sup.19R.sup.20).sub.nNR.sup.16C(.dbd.Y')R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.16C(.dbd.Y')OR.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.18C(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nNR.sup.17SO.sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.nOC(.dbd.Y')NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nOS(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.nOP(.dbd.Y')(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nOP(OR.sup.16)(OR.sup.17),
--(CR.sup.19R.sup.20).sub.nS(O)R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2R.sup.16,
--(CR.sup.19R.sup.20).sub.nS(O).sub.2NR.sup.16R.sup.17,
--(CR.sup.19R.sup.20).sub.nS(O)(OR.sup.16),
--(CR.sup.19R.sup.20).sub.n S(O).sub.2(OR.sup.16),
--(CR.sup.19R.sup.20).sub.nSC(.dbd.Y')R.sup.16,
--(CR.sup.19R.sup.20).sub.nSC(.dbd.Y')OR.sup.16,
--(CR.sup.19R.sup.20).sub.n SC(.dbd.Y')NR.sup.16R.sup.17, and
R.sup.21;
[0291] each R.sup.16, R.sup.17 and R.sup.18 is independently H,
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl, wherein
said alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or
heteroaryl is optionally substituted with one or more groups
selected from halo, CN, --OCF.sub.3, CF.sub.3, --NO.sub.2,
C.sub.1-C.sub.6 alkyl, --OH, --SH, --O(C.sub.1-C.sub.6 alkyl),
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0292] or R.sup.16 and R.sup.17 together with the nitrogen to which
they are attached form a 3-8 membered saturated, unsaturated or
aromatic ring having 0-2 heteroatoms selected from O, S and N,
wherein said ring is optionally substituted with one or more groups
selected from halo, CN, --OCF.sub.3, CF.sub.3, --NO.sub.2,
C.sub.1-C.sub.6 alkyl, --OH, --SH, --O(C.sub.1-C.sub.6 alkyl),
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0293] R.sup.19 and R.sup.20 are independently selected from H,
C.sub.1-C.sub.12 alkyl, --(CH.sub.2).sub.n-aryl,
--(CH.sub.2).sub.n-carbocyclyl, --(CH.sub.2).sub.n-heterocyclyl,
and --(CH.sub.2).sub.n-heteroaryl;
[0294] R.sup.21 is C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.8 alkenyl,
C.sub.2-C.sub.8 alkynyl, carbocyclyl, heterocyclyl, aryl, or
heteroaryl, wherein each member of R.sup.21 is optionally
substituted with one or more groups selected from halo, oxo, CN,
--OCF.sub.3, CF.sub.3, --NO.sub.2, C.sub.1-C.sub.6 alkyl, --OH,
--SH, --O(C.sub.1-C.sub.6 alkyl), --S(C.sub.1-C.sub.6 alkyl),
--NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --SO.sub.2(C.sub.1-C.sub.6 alkyl), --CO.sub.2H,
--CO.sub.2(C.sub.1-C.sub.6 alkyl), --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)(C.sub.1-C.sub.6
alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHSO.sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)SO.sub.2(C.sub.1-C.sub.6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2NH(C.sub.1-C.sub.6 alkyl), --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --OC(O)NH.sub.2, --OC(O)NH(C.sub.1-C.sub.6 alkyl),
--OC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --OC(O)O(C.sub.1-C.sub.6
alkyl), --NHC(O)NH(C.sub.1-C.sub.6 alkyl),
--NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.6 alkyl), and
--N(C.sub.1-C.sub.6 alkyl)C(O)O(C.sub.1-C.sub.6 alkyl);
[0295] each Y' is independently O, NR.sup.22, or S; and
[0296] R.sup.22 is H or C.sub.1-C.sub.12 alkyl.
[0297] In some variations, the MEK inhibitor compound of the
formula (III) has the formula (III-a) or (III-b):
##STR00027##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined for the formula (III) or as defined in
WO 2009/085983 A1, incorporated herein by reference.
[0298] In some embodiments, the MEK inhibitor compound of the
formula (III) is a compound selected from the compounds listed in
Table 1, or a pharmaceutically acceptable salt or solvate
thereof.
TABLE-US-00023 TABLE 1 Compound No. Chemical Name Structure (III)-5
5-(2-Fluoro-4- iodophenylamino)-imidazo[1,5-
a]pyridine-6-carboxylic acid (2- hydroxyethoxy)-amide ##STR00028##
(III)-6 5-(2-Fluoro-4-iodo- phenylamino)-imidazo[1,5-
a]pyridine-6-carboxylic acid ((R)-2,3-dihydroxy-propoxy)- amide
##STR00029## (III)-7 5-(2-Fluoro-4-iodo- phenylamino)-imidazo[1,5-
a]pyridine-6-carboxylic acid ((S)-2-hydroxy-propoxy)-amide
##STR00030## (III)-8 5-(4-Bromo-2- fluorophenylamino)-
imidazo[1,5-a]pyridine-6- carboxylic acid (2- hydroxyethoxy)-amide
##STR00031## (III)-9 5-(4-Bromo-2-fluoro- phenylamino)-imidazo[1,5-
a]pyridine-6-carboxylic acid ((S)-2-hydroxy-propoxy)-amide
##STR00032## (III)-10 5-(4-Bromo-2-fluoro- phenylamino)-8-fluoro-
imidazo[1,5-a]pyridine-6- carboxylic acid ((S)-2-hydroxy-
propoxy)-amide ##STR00033## (III)-11 8-Fluoro-5-(2-fluoro-4-iodo-
phenylamino)-imidazo[1,5- a]pyridine-6-carboxylic acid (2-
hydroxy-ethoxy)-amide ##STR00034## (III)-12
8-Fluoro-5-(2-fluoro-4-iodo- phenylamino)-imidazo[1,5-
a]pyridine-6-carboxylic acid ((R)-2,3-dihydroxy-propoxy)- amide
##STR00035## (III)-13 8-Fluoro-5-(2-fluoro-4-iodo-
phenylamino)-imidazo[1,5- a]pyridine-6-carboxylic acid
((S)-2-hydroxy-propoxy)-amide ##STR00036## (III)-14
5-(2-Fluoro-methanesulfanyl- phenylamino)-imidazo[1,5-
a]pyridine-6-carboxylic acid (2- hydroxy-ethoxy)-amide ##STR00037##
(III)-15 5-(2-Fluoro-4-iodo- phenylamino)-imidazo[1,5-
a]pyrazine-6-carboxylic acid (2- hydroxy-ethoxy)-amide ##STR00038##
(III)-16 5-(2-Fluoro-4-iodo- phenylamino)-imidazo[1,5-
a]pyrazine-6-carboxylic acid ((S)-2-hydroxy-propoxy)-amide
##STR00039## (III)-17 5-(4-Cyclopropyl-2-fluoro-
phenylamino)-imidazo[1,5- a]pyridine-6-carboxylic acid (2-
hydroxy-ethoxy)-amide ##STR00040## (III)-18
(R)-N-(2,3-Dihydroxypropoxy)- 5-(2-fluoro-4-
iodophenylamino)imidazo[1,5- a]pyrazine-6-carboxamide ##STR00041##
(III)-19 N-Ethoxy-5-(2-fluoro-4- iodophenylamino)imidazo[1,5-
a]pyrazine-6-carboxamide ##STR00042## (III)-20
N-(Cyclopropylmethoxy)-5-(2- fluoro-4- iodophenylamino)imidazo[1,5-
a]pyrazine-6-carboxamide ##STR00043## (III)-21 5-(2-Fluoro-4-
iodophenylamino)-N- methylimidazo[1,5-a]pyrazine- 6-carboxamide
##STR00044## (III)-22 5-(4-Bromo-2- fluorophenylamino)-N-(2-
hydroxy-ethoxy)imidazo[1,5- a]pyrazine-6-carboxamide ##STR00045##
(III)-23 (S)-5-(4-Bromo-2- fluorophenylamino)-N-(2-
hydroxy-propoxy)imidazo[1,5- a]pyrazine-6-carboxamide ##STR00046##
(III)-24 (R)-5-(4-Bromo-2- fluorophenylamino)-N-(2,3-
dihydroxy-propoxy)imidazo[1,5- a]pyrazine-6-carboxamide
##STR00047## (III)-25 5-(4-Bromo-2- fluorophenylamino)-N-
(cyclopropyl- methoxy)imidazo[1,5- a]pyrazine-6-carboxamide
##STR00048##
[0299] Compounds in Table 1 correspond to Examples 5-25 in WO
2009/085983 A1. Compounds (III)-5-(III)-20 and (III)-22-(III)-24
exhibited an IC.sub.50 of less than 0.5 .mu.M in the assay
described in Example 8b (MEK activity assay). Some of these
compounds exhibited an IC.sub.50 of less than 0.1 .mu.M. Compounds
(III)-21 and (III)-25 exhibited an IC.sub.50 of less than 10 .mu.M.
See page 49 in WO 2009/085983 A1.
[0300] Also embraced are MEK inhibitor compounds (and/or solvates
and salts thereof) described in WO 2009/085983 A1, which is
incorporated herein by reference, for example, imidazopyridine
compounds of the formula (III) (designated as formula I in WO
2009/085983 A1, e.g., on page 3) and variations thereof as
described in WO 2009/085983 A1. Compounds of formula (III) can be
synthesized using methods known in the art, for example, the
synthetic methods described in WO 2009/085983 A1, incorporated
herein by reference.
[0301] In some embodiments, the MEK inhibitor is a compound of
formula (IV),
##STR00049##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in WO 03/077914 A1 for the formula I
on pages 4-9 or any applicable variations described in WO 03/077914
A1, incorporated herein by reference.
[0302] In some variations, the MEK inhibitor compound of the
formula (IV) is a compound of the formula (IV-a), (IV-b), (IV-c),
or (IV-d):
##STR00050##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in WO 03/077914 A1 for the formulae
II, III, IIIa and IIIb, respectively on pages 10-13 or any
applicable variations described in WO 03/077914 A1, incorporated
herein by reference.
[0303] In some embodiments, the MEK inhibitor compound of the
formula (IV) is a compound selected from the group consisting of:
[0304]
7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-3H-benzoimidazole-5-carboxylic
acid cyclopropylmethoxy-amide; [0305]
6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylic
acid cyclopropylmethoxy-amide; [0306]
6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-ca-
rboxylic acid (2-hydroxy-ethoxy)-amide; [0307]
6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-ca-
rboxylic acid (2,3-dihydroxy-propoxy)-amide; [0308]
6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-
-3H-benzoimidazole-5-carboxylic acid (2-hydroxy-ethoxy)-amide;
[0309]
[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-bro-
mo-2-methyl-phenyl)-amine; [0310]
1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazol-5--
yl]-2-hydroxy-ethanone; [0311]
1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-met-
hoxy ethanone; [0312]
6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-ca-
rboxylic acid (2-hydroxy-1,1-dimethyl-ethoxy)-amide; [0313]
6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-furan-2-ylmethyl)-
-3H-benzoimidazole-5-carboxylic acid (2-hydroxy-ethoxy)-amide;
[0314]
6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylic
acid (2-hydroxy-ethoxy)-amide; [0315]
6-(-Bromo-2-fluoro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-car-
boxylic acid (2-hydroxy-ethoxy)-amide; and [0316]
6-(2,4-Dichloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbox-
ylic acid (2-hydroxy-ethoxy)-amide; [0317] or a pharmaceutically
acceptable salt or solvate thereof.
[0318] Also embraced are any variations of formula (IV) as
described in WO 03/077914 A1, which is incorporated herein by
reference. Compounds of the formula (IV) or any variations thereof
can be synthesized using methods known in the art, for example, the
synthetic methods described in WO 03/077914 A1, incorporated herein
by reference.
[0319] In some embodiments, the MEK inhibitor is a compound of
formula (V),
##STR00051##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in WO 2005/121142 A1 for the formula
[I] on pages 6-10 or any applicable variations described in WO
2005/121142 A1, incorporated herein by reference.
[0320] Also embraced are any variations of formula (V) as described
in WO 2005/121142 A1, such as the individual MEK inhibitor
compounds described in WO 2005/121142 A1, e.g., Examples 1-1 to
1-343 in Table 1, Examples 2-1 and 2-2 in Table 2, Examples 3-1 to
3-9 in Table 3, Examples 4-1 to 4-148 in Table 4. Compounds of the
formula (V) or any variations thereof can be synthesized using
methods known in the art, for example, the synthetic methods
described in WO 2005/121142 A1, incorporated herein by
reference.
[0321] In some embodiments, the MEK inhibitor is a compound of
formula (VI),
##STR00052##
or a pharmaceutically acceptable salt or ester thereof,
wherein:
[0322] R1 is selected from the group consisting of bromo, iodo,
ethynyl, cycloalkyl, alkoxy, azetidinyl, acetyl, heterocycyl,
cyano, straight-chained alkyl and branched-chain alkyl;
[0323] R2 is selected from the group consisting of hydrogen,
chlorine, fluorine, and alkyl;
[0324] R3 is selected from the group consisting of hydrogen,
chlorine, and fluorine;
[0325] R4 is selected from the group consisting of hydrogen,
optionally substituted aryl, alkyl, and cycloalkyl;
[0326] R5 is selected from the group consisting of hydrogen and
##STR00053##
[0327] wherein R6 is selected from the group consisting of
hydroxyl, alkoxy, cycloalkyl, optionally substituted alkyl,
optionally substituted aryl, and optionally substituted
heteroaryl;
[0328] R7 and R8 are independently selected from the group
consisting of hydrogen and optionally substituted alkyl;
[0329] or R6 and R7 can together form a cycloalkyl group and R8 is
hydrogen.
[0330] In some variations, the MEK inhibitor compound is of the
formula (VI), or a pharmaceutically acceptable salt or ester
thereof, wherein the variables are as defined in WO 2007/096259 A1
for the formula I or any applicable variations described on pages
4-10 in WO 2007/096259 A1, incorporated herein by reference.
Further embraced MEK inhibitors are compounds described in Examples
1-182 in WO 2007/096259 A1, incorporated herein by reference.
[0331] In some embodiments, the MEK inhibitor compound of the
formula (VI) is a compound selected from the group consisting of:
[0332]
(2S,3S)-N-(4-Bromo-phenyl)-2-[(R)-4-(4-methoxy-phenyl)-2,5-dioxo-imidazol-
idin-1-yl]-3-phenyl-butyramide; [0333]
(2S,3S)-N-(4-Iodo-phenyl)-2-[(R)-4-(4-methoxy-phenyl)-2,5-dioxo-imidazoli-
din-1-yl]-3-phenyl-butyramide; [0334]
(2S,3S)-N-(2-Fluoro-4-iodo-phenyl)-2-{(R)-4-[4-(2-hydroxy-ethoxy)-phenyl]-
-2,5-dioxo-imidazolidin-1-yl}-3-phenyl-butyramide; [0335]
(2S,3S)-N-(4-Ethynyl-2-fluoro-phenyl)-2-{(R)-4-[4-(2-hydroxy-ethoxy)-phen-
yl]-2,5-dioxo-imidazolidin-1-yl}-3-phenyl-butyramide; [0336]
(2R,3S)-N-(4-Ethynyl-2-fluoro-phenyl)-2-{(R)-4-[4-(2-hydroxy-ethoxy)-phen-
yl]-2,5-dioxo-imidazolidin-1-yl}-3-phenyl-butyramide; [0337]
(2S,3S)-N-(2-Chloro-4-iodo-phenyl)-2-{(R)-4-[4-(2-hydroxy-ethoxy)-phenyl]-
-2,5-dioxo-imidazolidin-1-yl}-3-phenyl-butyramide; [0338]
(2S,3S)-2-{(R)-4-[4-(2-Hydroxy-ethoxy)-phenyl]-2,5-dioxo-imidazolidin-1-y-
l}-N-(4-iodo-2-methyl-phenyl)-3-phenyl-butyramide; [0339]
(2S,3S)-N-(2-Chloro-4-iodo-phenyl)-2-{(R)-4-[4-((R)-2,3-dihydroxy-propoxy-
)-phenyl]-2,5-dioxo-imidazolidin-1-yl}-3-phenyl-butyramide; [0340]
(2S,3S)-N-(2-Chloro-4-iodo-phenyl)-2-{(R)-4-[4-((S)-2,3-di
hydroxy-propoxy)-phenyl]-2,5-dioxo-imidazolidin-1-yl}-3-phenyl-butyramide-
; [0341]
(2S,3S)-2-{(R)-2,5-Dioxo-4-[4-(2-oxo-2-pyrrolidin-1-yl-ethoxy)-ph-
enyl]-imidazolidin-1-yl}-N-(2-fluoro-4-iodo-phenyl)-3-phenyl-butyramide;
[0342]
(2S,3S)-2-((R)-2,5-Dioxo-4-thiophen-3-yl-imidazolidin-1-yl)-N-(4-i-
odo-phenyl)-3-phenyl-butyramide; [0343]
(S)-2-[(R)-4-(2,3-Dihydro-benzo[1,4]dioxin-6-yl)-2,5-dioxo-imidazolidin-1-
-yl]-N-(2-fluoro-4-iodo-phenyl)-3-phenyl-propionamide; [0344]
(S)-2-[(R)-4-(4-Acetylamino-phenyl)-2,5-dioxo-imidazolidin-1-yl]-N-(2-flu-
oro-4-iodo-phenyl)-3-phenyl-propionamide; [0345]
(4-{(R)-1-[(1S,2S)-1-(2-Fluoro-4-iodo-phenylcarbamoyl)-2-phenyl-propyl]-2-
,5-dioxo-imidazolidin-4-yl}-phenoxymethyl)-phosphonic acid dimethyl
ester; [0346]
(2S,3S)-N-(2-Fluoro-4-iodo-phenyl)-2-((R)-4-isopropyl-2,5-dioxo-im-
idazolidin-1-yl)-3-phenyl-butyramide; [0347]
(S)-N-(2-Fluoro-4-iodo-phenyl)-2-{(R)-4-[4-(2-hydroxy-ethoxy)-phenyl]-2,5-
-dioxo-imidazolidin-1-yl}-3-methyl-butyramide; [0348]
(S)-N-(2-Fluoro-4-iodo-phenyl)-2-[(R)-4-(4-methoxy-phenyl)-2,5-dioxo-imid-
azolidin-1-yl]-3-o-tolyl-propionamide; [0349]
(S)-N-(2-Fluoro-4-iodo-phenyl)-2-[(R)-4-(4-methoxy-phenyl)-2,5-dioxo-imid-
azolidin-1-yl]-3-m-tolyl-propionamide; [0350]
(S)-N-(2-Fluoro-4-iodo-phenyl)-2-[(R)-4-(4-methoxy-phenyl)-2,5-dioxo-imid-
azolidin-1-yl]-3-p-tolyl-propionamide; and [0351]
(S)-N-(4-Cyclopropyl-2-fluoro-phenyl)-3-(4-fluoro-phenyl)-2-{(R)-4-[4-(2--
hydroxy-1-hydroxymethyl-ethoxy)-phenyl]-2,5-dioxo-imidazolidin-1-yl}-propi-
onamide; [0352] or a pharmaceutically acceptable salt or ester
thereof.
[0353] In some embodiments, the MEK inhibitor is a compound of
formula (VII),
##STR00054##
or a pharmaceutically acceptable salt or ester thereof,
wherein:
[0354] R1 is selected from the group consisting of halogen,
ethynyl, and cycloalkyl;
[0355] R2 is selected from the group consisting of hydrogen and
CH(R3)(R4);
[0356] R3 is selected from the group consisting of lower alkyl,
lower alkoxy, optionally substituted aryl, and optionally
substituted heteroaryl;
[0357] R4 is selected from the group consisting of hydrogen and
lower alkyl;
[0358] R5 is hydrogen or, taken together with R2 and the carbon to
which R2 and R5 are attached, forms lower cycloalkyl; and
[0359] R6 is selected from the group consisting of hydrogen, lower
alkyl, lower cycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl.
[0360] In some variations, the MEK inhibitor compound is of the
formula (VI), or a pharmaceutically acceptable salt or ester
thereof, wherein the variables are as defined in WO 2009/021887 A1
for the formula I or any applicable variations described on pages
4-5 in WO 2009/021887 A1, incorporated herein by reference. Further
embraced MEK inhibitors are compounds described in Examples 1-21 in
2009/021887 A1, incorporated herein by reference.
[0361] In some embodiments, the MEK inhibitor compound of the
formula (VI) is a compound selected from the group consisting of:
[0362]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-[(S)-1-(6-iodo-1H-benzoimidazol-2-y-
l)-2-phenyl-ethyl]-imidazolidine-2,4-dione; [0363]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-(5-iodo-1H-benzoimidazol-2-ylmethyl-
)-imidazolidine-2,4-dione; [0364]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-[(S)-1-(5-iodo-1H-benzoimidazol-2-y-
l)-2-methyl-propyl]-imidazolidine-2,4-dione; [0365]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-[(1R,2R)-1-(5-iodo-1H-benzoimidazol-
-2-yl)-2-methoxy-propyl]-imidazolidine-2,4-dione; [0366]
3-[(S)-1-(5-Iodo-1H-benzoimidazol-2-yl)-2-phenyl-ethyl]-imidazolidine-2,4-
-dione; compound with trifluoro-acetic acid; [0367]
(R)-3-[(S)-2-(4-Fluoro-phenyl)-1-(5-iodo-1H-benzoimidazol-2-yl)-ethyl]-5--
[4-(2-hydroxy-ethoxy)-phenyl]-imidazolidine-2,4-dione; [0368]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-[(S)-1-(5-iodo-1H-benzoimidazol-2-y-
l)-2-(4-methoxy-phenyl)-ethyl]-imidazolidine-2,4-dione; [0369]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-[(S)-1-(5-iodo-1H-benzoimidazol-2-y-
l)-2-thiophen-2-yl-ethyl]-imidazolidine-2,4-dione; [0370]
(R)-3-[(1S,2S)-1-(6-Iodo-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-5-phenyl-
-imidazolidine-2,4-dione; [0371]
(R)-3-[(1S,2S)-1-(6-Iodo-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-5-(4-met-
hoxy-phenyl)-imidazolidine-2,4-dione; [0372]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-[(1S,2S)-1-(6-iodo-1H-benzoimidazol-
-2-yl)-2-phenyl-propyl]-imidazolidine-2,4-dione; [0373]
(R)-3-[(1S,2S)-1-(6-Iodo-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-5-[4-(2--
methoxy-ethoxy)-phenyl]-imidazolidine-2,4-dione; [0374]
2-(4-{(R)-1-[(1S,2S)-1-(6-Iodo-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-2,-
5-dioxo-imidazolidin-4-yl}-phenoxy)-N,N-dimethyl-acetamide; [0375]
N,N-Bis-(2-hydroxy-ethyl)-2-(4-{(R)-1-[(1S,2S)-1-(6-iodo-1H-benzoimidazol-
-2-yl)-2-phenyl-propyl]-2,5-dioxo-imidazolidin-4-yl}-phenoxy)-acetamide;
[0376]
(R)-3-[(1S,2S)-1-(5-Iodo-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-5-
-isopropyl-imidazolidine-2,4-dione; [0377] (R)-5-Cyclohexyl-3-[(1
S,2S)-1-(5-iodo-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-imidazolidine-2,4-
-dione; [0378]
(R)-5-[4-(2-Hydroxy-ethoxy)-phenyl]-3-[1-(5-iodo-1H-benzoimidazol-2-yl)-c-
yclopropyl]-imidazolidine-2,4-dione; [0379]
(R)-3-[(1S,2S)-1-(6-Bromo-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-5-[4-(2-
-hydroxy-ethoxy)-phenyl]-imidazolidine-2,4-dione; [0380]
(R)-3-[(S)-1-(5-Cyclopropyl-1H-benzoimidazol-2-yl)-2-phenyl-ethyl]-5-[4-(-
2-hydroxy-ethoxy)-phenyl]-imidazolidine-2,4-dione; [0381]
(R)-3-[(S)-1-(5-Ethynyl-1H-benzoimidazol-2-yl)-2-phenyl-ethyl]-5-[4-(2-hy-
droxy-ethoxy)-phenyl]-imidazolidine-2,4-dione; and [0382]
(R)-3-[(1S,2S)-1-(5-Ethynyl-1H-benzoimidazol-2-yl)-2-phenyl-propyl]-5-[4--
(2-hydroxy-ethoxy)-phenyl]-imidazolidine-2,4-dione; [0383] or a
pharmaceutically acceptable salt or solvate thereof.
[0384] In some embodiments, the MEK inhibitor is a compound
selected from the group consisting of GDC-0973 (Methanone,
[3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl][3-hydroxy-3-(2S)-2--
piperidinyl-1-azetidinyl]-), G-38963, G02443714, G02442104, and
G00039805, or a pharmaceutically acceptable salt or solvate
thereof.
##STR00055##
IV Kits
[0385] In another aspect, provided herein is a kit comprising a
PD-L1 axis binding antagonist and/or a MEK inhibitor for treating
or delaying progression of a cancer in an individual or for
enhancing immune function of an individual having cancer. In some
embodiments, the kit comprises a PD-1 axis binding antagonist and a
package insert comprising instructions for using the PD-1 axis
binding antagonist in combination with a MEK inhibitor to treat or
delay progression of cancer in an individual or to enhance immune
function of an individual having cancer. In some embodiments, the
kit comprises a MEK inhibitor and a package insert comprising
instructions for using the MEK inhibitor in combination with a PD-1
axis binding antagonist to treat or delay progression of cancer in
an individual or to enhance immune function of an individual having
cancer. In some embodiments, the kit comprises a PD-1 axis binding
antagonist and a MEK inhibitor, and a package insert comprising
instructions for using the PD-1 axis binding antagonist and the MEK
inhibitor to treat or delay progression of cancer in an individual
or to enhance immune function of an individual having cancer. Any
of the PD-1 axis binding antagonists and/or MEK inhibitors
described herein may be included in the kits.
[0386] In some embodiments, the kit comprises a container
containing one or more of the PD-1 axis binding antagonists and MEK
inhibitors described herein. Suitable containers include, for
example, bottles, vials (e.g., dual chamber vials), syringes (such
as single or dual chamber syringes) and test tubes. The container
may be formed from a variety of materials such as glass or plastic.
In some embodiments, the kit may comprise a label (e.g., on or
associated with the container) or a package insert. The label or
the package insert may indicate that the compound contained therein
may be useful or intended for treating or delaying progression of
cancer in an individual or for enhancing immune function of an
individual having cancer. The kit may further comprise other
materials desirable from a commercial and user standpoint,
including other buffers, diluents, filters, needles, and
syringes.
EXAMPLES
[0387] The invention can be further understood by reference to the
following examples, which are provided by way of illustration and
are not meant to be limiting.
Example 1: Combination Treatment with an Anti-PDL1 Antibody and a
MEK Inhibitor Causes Sustained Tumor Regression in
Vemurafenib-Progressing Tumors
[0388] While B-raf inhibition (such as by treatment with
Vemurafenib) is effective in eliciting short-term tumor regression,
resistance is frequently observed. This Example describes the
finding that treatment with a combination of a PD-1 axis binding
antagonist and a MEK inhibitor induces sustained tumor regression
and increased progression-free survival in animals with
Vemurafenib-progressing tumors. Moreover, treatment with a
combination of a PD-1 axis binding antagonist and a MEK inhibitor
was surprisingly superior to treatment with either agent
individually.
[0389] Materials and Methods
Mouse Model
[0390] A melanoma GEM model B-raf.sup.V600E;PTEN.sup.fl/fl; TyCreER
was used. B-raf.sup.V600E and TyCreER alleles were as described in
Dankort, D., et al Nat. Genet. 41(5):544-52 (2009). The PTEN
conditional allele was as described in Lesche, R. et al. genesis
32:148-9 (2002).
Tumor Initiation
[0391] Tumors were initiated by application of tamoxifen as
described in Dankort, D., et al. Nat. Genet. 41(5):544-52 (2009).
Animals were enrolled into the study once their tumors reached a
size greater than or equal to 400 mm.sup.3.
Treatments
[0392] Prior to beginning treatment, each mouse received a biopsy
of a melanoma tumor. After the biopsy, mice were allowed to recover
for up to one week prior to receiving treatment. Mice were assigned
into initial treatment groups (n=20), and treatment commenced at
day 0.
[0393] For Vemurafenib treatment, mice were given either MCT, 200
.mu.L, PO, qd; or PLX-4032 (Vemurafenib), 50 mg/kg PO, BID (volume
not to exceed 300 .mu.L). When animals in any group reached
.about.2000 mm.sup.3, the tumors were biopsied a second time. After
the biopsy procedure, mice recovered for up to one week prior to
receiving further therapeutic treatment. The animals were then
re-assigned to the following treatment groups: GDC-0973
(Cobimetinib), 7.5 mg/kg PO, qd (volume not to exceed 300
.mu.L)+Ragweed Control (IgG2a), 10 mg/kg IP, three times weekly;
GDC-0973, 7.5 mg/kg PO, qd (volume not to exceed 300
.mu.L)+anti-PDL1 (IgG1-WT), 10 mg/kg IP, three times weekly; or
MCT, 200 uL, PO, qd+anti-PDL1 (IgG1-WT), 10 mg/kg IP, three times
weekly. IP dose volume did not exceed 300 .mu.L.
[0394] Mice were weighed and tumors measured at least once a week
until study termination. Mice received treatment each day until a
mean tumor volume of 2500 mm.sup.3 was achieved. Mice were then
euthanized, and melanoma tumors were collected for histology and
assessing molecular changes. Mice were perfused under anesthesia at
euthanasia.
[0395] Throughout the study, mice were monitored for clinical
appearance (body condition, coat appearance, posture, labored
breathing, etc.) at least 2 times a week, with increasing
frequency, up to daily, depending on severity of adverse clinical
signs observed. Moribund animals were euthanized. Mice with a body
condition score <2 were euthanized.
[0396] Results
[0397] Upon tumor induction, the melanoma GEM model
B-raf.sup.V600E;PTEN.sup.fl/fl;TyCreER causes tumors that show an
initial regression in size upon treatment with the B-raf inhibitor
Vemurafenib. After this initial regression, the tumors display
steady re-growth, thereby modeling resistance to B-raf inhibition
in Vemurafenib-progressing tumors.
[0398] This model was used to test the efficacy of PD-1 axis
binding antagonists and MEK inhibitors as a 2.sup.nd line therapy
for Vemurafenib-progressing tumors. As shown in FIG. 1, after first
line treatment with Vemurafenib, animals were treated with an
antibody against PD-L1, a MEK inhibitor (Cobimetinib), or both.
Treatment with anti-PD-L1 alone showed no effect on tumor growth.
Treatment with Cobimetinib caused an initial tumor regression, but
this response was not sustained and tumor re-growth was observed.
Combination treatment with anti-PD-L1 and Cobimetinib, however,
caused regression in every tumor, and this regression was
sustained. In addition, intratumoral GR1 levels were significantly
reduced, and a signature of T cell activation was observed (e.g.,
increased CD8, PRF1, and MHC I). Importantly, treatment with
anti-PD-L1 and Cobimetinib led to increased progression-free
survival (PFS).
[0399] FIG. 2 shows individual animal responses following crossover
from Vemurafenib to anti-PD-L1, Cobimetinib, or combination
treatment.
[0400] These results demonstrate that combined treatment with a
PD-1 axis binding antagonist and a MEK inhibitor leads to dramatic,
sustained tumor regression in Vemurafenib-progressing tumors, as
compared to treatment with each agent alone. Moreover, these
results demonstrate the superior efficacy of combined PD-1 axis/MEK
inhibition as a 2.sup.nd line treatment for tumors resistant to
B-raf inhibition.
[0401] All patents, patent applications, documents, and articles
cited herein are herein incorporated by reference in their
entireties.
Sequence CWU 1
1
28110PRTArtificial SequenceSynthetic ConstructVARIANT6Xaa = D or G
1Gly Phe Thr Phe Ser Xaa Ser Trp Ile His1 5 10218PRTArtificial
SequenceSynthetic ConstructVARIANT4Xaa = S or LVARIANT10Xaa = T or
S 2Ala Trp Ile Xaa Pro Tyr Gly Gly Ser Xaa Tyr Tyr Ala Asp Ser Val1
5 10 15Lys Gly39PRTArtificial SequenceSynthetic Construct 3Arg His
Trp Pro Gly Gly Phe Asp Tyr1 5425PRTArtificial SequenceSynthetic
Construct 4Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro
Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser 20
25513PRTArtificial SequenceSynthetic Construct 5Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val1 5 10632PRTArtificial
SequenceSynthetic Construct 6Arg Phe Thr Ile Ser Ala Asp Thr Ser
Lys Asn Thr Ala Tyr Leu Gln1 5 10 15Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys Ala Arg 20 25 30711PRTArtificial
SequenceSynthetic Construct 7Trp Gly Gln Gly Thr Leu Val Thr Val
Ser Ala1 5 10811PRTArtificial SequenceSynthetic
ConstructVARIANT5Xaa = D or VVARIANT6Xaa = V or IVARIANT7Xaa = S or
NVARIANT9Xaa = A or FVARIANT10Xaa = V or L 8Arg Ala Ser Gln Xaa Xaa
Xaa Thr Xaa Xaa Ala1 5 1097PRTArtificial SequenceSynthetic
ConstructVARIANT4Xaa = F or TVARIANT6Xaa = Y or A 9Ser Ala Ser Xaa
Leu Xaa Ser1 5109PRTArtificial SequenceSynthetic
ConstructVARIANT3Xaa = Y, G, F, or SVARIANT4Xaa = L, Y, F or
WVARIANT5Xaa = Y, N, A, T, G, F or IVARIANT6Xaa = H, V, P, T or
IVARIANT8Xaa = A, W, R, P or T 10Gln Gln Xaa Xaa Xaa Xaa Pro Xaa
Thr1 51123PRTArtificial SequenceSynthetic Construct 11Asp Ile Gln
Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg
Val Thr Ile Thr Cys 201215PRTArtificial SequenceSynthetic Construct
12Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr1 5 10
151332PRTArtificial SequenceSynthetic Construct 13Gly Val Pro Ser
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr1 5 10 15Leu Thr Ile
Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 20 25
301411PRTArtificial SequenceSynthetic Construct 14Phe Gly Gln Gly
Thr Lys Val Glu Ile Lys Arg1 5 101510PRTArtificial
SequenceSynthetic Construct 15Gly Phe Thr Phe Ser Asp Ser Trp Ile
His1 5 101618PRTArtificial SequenceSynthetic Construct 16Ala Trp
Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val1 5 10 15Lys
Gly1711PRTArtificial SequenceSynthetic Construct 17Arg Ala Ser Gln
Asp Val Ser Thr Ala Val Ala1 5 10187PRTArtificial SequenceSynthetic
Construct 18Ser Ala Ser Phe Leu Tyr Ser1 5199PRTArtificial
SequenceSynthetic Construct 19Gln Gln Tyr Leu Tyr His Pro Ala Thr1
520118PRTArtificial SequenceSynthetic Construct 20Glu Val Gln Leu
Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser 20 25 30Trp Ile
His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala
Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55
60Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr65
70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
Cys 85 90 95Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln
Gly Thr 100 105 110Leu Val Thr Val Ser Ala 11521108PRTArtificial
SequenceSynthetic Construct 21Asp Ile Gln Met Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg
Ala Ser Gln Asp Val Ser Thr Ala 20 25 30Val Ala Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ser Ala Ser Phe Leu
Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe
Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Tyr His Pro Ala 85 90 95Thr Phe
Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 10522440PRTArtificial
SequenceSynthetic Construct 22Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Asp Cys Lys Ala
Ser Gly Ile Thr Phe Ser Asn Ser 20 25 30Gly Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp
Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Thr
Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105
110Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser
115 120 125Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val
Lys Asp 130 135 140Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
Gly Ala Leu Thr145 150 155 160Ser Gly Val His Thr Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr 165 170 175Ser Leu Ser Ser Val Val Thr
Val Pro Ser Ser Ser Leu Gly Thr Lys 180 185 190Thr Tyr Thr Cys Asn
Val Asp His Lys Pro Ser Asn Thr Lys Val Asp 195 200 205Lys Arg Val
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala 210 215 220Pro
Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro225 230
235 240Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
Val 245 250 255Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn
Trp Tyr Val 260 265 270Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
Pro Arg Glu Glu Gln 275 280 285Phe Asn Ser Thr Tyr Arg Val Val Ser
Val Leu Thr Val Leu His Gln 290 295 300Asp Trp Leu Asn Gly Lys Glu
Tyr Lys Cys Lys Val Ser Asn Lys Gly305 310 315 320Leu Pro Ser Ser
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 325 330 335Arg Glu
Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr 340 345
350Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
355 360 365Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
Asn Tyr 370 375 380Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
Phe Phe Leu Tyr385 390 395 400Ser Arg Leu Thr Val Asp Lys Ser Arg
Trp Gln Glu Gly Asn Val Phe 405 410 415Ser Cys Ser Val Met His Glu
Ala Leu His Asn His Tyr Thr Gln Lys 420 425 430Ser Leu Ser Leu Ser
Leu Gly Lys 435 44023214PRTHomo sapiens 23Glu Ile Val Leu Thr Gln
Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30Leu Ala Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala
Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75
80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Asn Trp Pro Arg
85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala
Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr
Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200
205Phe Asn Arg Gly Glu Cys 21024118PRTArtificial SequenceSynthetic
Construct 24Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro
Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
Ser Asp Ser 20 25 30Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr
Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr
Ser Lys Asn Thr Ala Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg His Trp Pro Gly
Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser
Ser 1152511PRTArtificial SequenceSynthetic Construct 25Trp Gly Gln
Gly Thr Leu Val Thr Val Ser Ser1 5 1026448PRTArtificial
SequenceSynthetic Construct 26Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Asp Ser 20 25 30Trp Ile His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Trp Ile Ser Pro Tyr
Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105
110Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly 130 135 140Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn145 150 155 160Ser Gly Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln 165 170 175Ser Ser Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205Asn Thr Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220His
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser225 230
235 240Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg 245 250 255Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro 260 265 270Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala 275 280 285Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Ala Ser Thr Tyr Arg Val Val 290 295 300Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr305 310 315 320Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345
350Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
Glu Ser 370 375 380Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val Leu Asp385 390 395 400Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp Lys Ser 405 410 415Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His Glu Ala 420 425 430Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440
44527214PRTArtificial SequenceSynthetic Construct 27Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Ser Thr Ala 20 25 30Val Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr
Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Tyr His Pro
Ala 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val
Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn
Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn
Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu
Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200
205Phe Asn Arg Gly Glu Cys 21028122PRTArtificial SequenceSynthetic
Construct 28Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro
Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
Ser Asp Ser 20 25 30Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr
Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr
Ser Lys Asn Thr Ala Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg His Trp Pro Gly
Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser
Ser Ala Ser Thr Lys 115 120
* * * * *