U.S. patent application number 12/903051 was filed with the patent office on 2011-07-07 for compositions and methods using anti-cs1 antibodies to treat multiple myeloma.
This patent application is currently assigned to Facet Biotech Corporation. Invention is credited to Daniel Afar.
Application Number | 20110165154 12/903051 |
Document ID | / |
Family ID | 39033619 |
Filed Date | 2011-07-07 |
United States Patent
Application |
20110165154 |
Kind Code |
A1 |
Afar; Daniel |
July 7, 2011 |
COMPOSITIONS AND METHODS USING ANTI-CS1 ANTIBODIES TO TREAT
MULTIPLE MYELOMA
Abstract
Compositions and methods for treating MM are provided
herein.
Inventors: |
Afar; Daniel; (Fremont,
CA) |
Assignee: |
Facet Biotech Corporation
Redwood City
CA
|
Family ID: |
39033619 |
Appl. No.: |
12/903051 |
Filed: |
October 12, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11835245 |
Aug 7, 2007 |
7842293 |
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12903051 |
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60836250 |
Aug 7, 2006 |
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60856144 |
Nov 1, 2006 |
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Current U.S.
Class: |
424/133.1 ;
435/325 |
Current CPC
Class: |
A61K 31/454 20130101;
A61P 25/02 20180101; A61K 33/24 20130101; A61P 43/00 20180101; A61K
9/127 20130101; A61K 9/0019 20130101; A61K 2039/505 20130101; A61K
2039/545 20130101; A61K 31/704 20130101; A61K 31/475 20130101; A61K
31/675 20130101; A61P 35/00 20180101; A61K 45/06 20130101; C07K
16/2896 20130101; A61P 37/04 20180101; A61K 2039/507 20130101; A61K
39/39558 20130101; A61K 31/573 20130101; A61K 31/7048 20130101;
A61P 7/00 20180101; A61P 25/00 20180101; C07K 16/22 20130101; C07K
16/2803 20130101; A61K 31/454 20130101; A61K 2300/00 20130101; A61K
31/475 20130101; A61K 2300/00 20130101; A61K 31/573 20130101; A61K
2300/00 20130101; A61K 31/704 20130101; A61K 2300/00 20130101; A61K
39/39558 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/133.1 ;
435/325 |
International
Class: |
A61K 39/395 20060101
A61K039/395; C12N 5/09 20100101 C12N005/09; A61P 37/04 20060101
A61P037/04; A61P 35/00 20060101 A61P035/00 |
Claims
1. A pharmaceutical composition suitable for treating multiple
myeloma in a subject, comprising a therapeutically effective amount
of HuLuc63 in combination with one or more therapeutic agents,
wherein said composition is capable of being administered in a
single or multiple dose regimen.
2. A pharmaceutical composition suitable for treating multiple
myeloma in a subject, comprising a therapeutically effective amount
of HuLuc63 in combination with a one or more targeted agents,
conventional chemotherapeutic agents, hormonal therapy agents, or
supportive care agents, and/or combinations thereof, wherein said
composition is capable of being administered in a single or
multiple dose regimen.
3. The pharmaceutical composition according to claim 2, in which
one or more conventional therapeutic agents are selected from the
group consisting of thalidomide, vincristine, carmustine,
melphalan, cyclophosphamide, doxorubicin, doxorubicin HCl liposome
injection, cisplatin, etoposide, ara-C, and one or more hormonal
therapy agents selected from the group consisting of dexamethasone
and prednisone.
4. The pharmaceutical composition according to claim 2, in which
one or more one or more targeted agents are selected from the group
consisting of anti-angiogenic agents, inhibitors of growth factor
signaling, immunomodulators, inhibitors of protein synthesis,
folding and/or degradation, inhibitors of gene expression,
pro-apoptotic agents, agents that inhibit signal transduction,
other agents, and combinations thereof,
5. The pharmaceutical composition according to claim 3, comprising
dexamethasone.
6. The pharmaceutical composition according to claim 3, comprising
dexamethasone and thalidomide.
7. The pharmaceutical composition according to claim 3, comprising
vincristine, doxorubicin and dexamethasone.
8. The pharmaceutical composition according to claim 3, comprising
doxorubicin HCl liposome injection, vincristine and
dexamethasone.
9. The pharmaceutical composition according to claim 3, comprising
cyclophosphamide.
10. The pharmaceutical composition according to claim 3, comprising
dexamethasone, thalidomide, cisplatin, doxorubicin,
cyclophosphamide and etoposide.
11. The pharmaceutical composition according to claim 3, comprising
vincristine, doxorubicin, dexamethasone, and cyclophosphamide.
12. The pharmaceutical composition according to claim 3, comprising
doxorubicin HCl liposome injection, vincristine, dexamethasone, and
thalidomide.
13. The pharmaceutical composition according to claim 1 or 2, in
which HuLuc63 is in an injectable form.
14. The pharmaceutical composition according to claim 12,
containing from about 0.5 mg/kg to about 20 mg/kg of HuLuc63.
15. The pharmaceutical composition according to claim 1 or 2, in
which administration of said pharmaceutical composition elicits a
complete response.
16. The pharmaceutical composition according to claim 1 or 2, in
which administration of said pharmaceutical composition elicits a
very good partial response.
17. The pharmaceutical composition according to claim 1 or 2, in
which administration of said pharmaceutical composition elicits a
partial response.
18. The pharmaceutical composition according to claim 1 or 2, in
which administration of said pharmaceutical composition elicits a
minimal response.
19. A method for increasing sensitivity of a multiple myeloma cell
to a therapeutic agent, comprising contacting the cell with a
therapeutically effective amount of HuLuc63.
20. The method according to claim 19, further comprising contacting
the cell with a therapeutically effective amount of one or more
conventional therapeutic agents selected from the group consisting
of thalidomide, vincristine, carmustine, melphalan,
cyclophosphamide, doxorubicin, doxorubicin HCl liposome injection,
cisplatin, etoposide, ara-C, and one or more hormonal therapy
agents selected from the group consisting of dexamethasone and
prednisone.
21. The method according to claim 19, wherein the multiple myeloma
cell is resistant to the therapeutic agent.
22. The method according to claim 21, further comprising contacting
said resistant cell with a therapeutic agent that the cell is
sensitive to, said agent selected from the group consisting of one
or more conventional therapeutic agents selected from the group
consisting of thalidomide, vincristine, carmustine, melphalan,
cyclophosphamide, doxorubicin, doxorubicin HCl liposome injection,
cisplatin, etoposide, ara-C, and one or more hormonal therapy
agents selected from the group consisting of dexamethasone and
prednisone.
23. A method of treating multiple myeloma in a subject, the method
comprising administering a first pharmaceutical composition
comprising HuLuc63 and a second pharmaceutical composition
comprising one or more targeted agents, conventional
chemotherapeutic agents, hormonal therapy agents, or supportive
care agents, or combinations thereof, wherein said compositions are
capable of being administered in single or multiple dose
regimens.
24. A method of treating multiple myeloma in a subject, the method
comprising administering a therapeutically effective amount of
HuLuc63 in combination with one or more targeted agents selected
from the group consisting of anti-angiogenic agents, inhibitors of
growth factor signaling, immunomodulators, inhibitors of protein
synthesis, folding and/or degradation, inhibitors of gene
expression, pro-apoptotic agents, agents that inhibit signal
transduction, other agents, or combinations thereof, wherein said
compositions are capable of being administered in single or
multiple dose regimens.
25. The method according to claim 24, further comprising one or
more conventional chemotherapeutic agents and one or more hormonal
agents.
26. The method according to claim 25, in which one or more
conventional therapeutic agents are selected from the group
consisting of thalidomide, vincristine, carmustine, melphalan,
cyclophosphamide, doxorubicin, doxorubicin HCl liposome injection,
cisplatin, etoposide, ara-C, and one or more hormonal therapy
agents selected from the group consisting of dexamethasone and
prednisone.
27. The method according to claim 23, in which HuLuc63 is
administered intravenously at a dosage from approximately 0.5 mg/kg
to approximately 20 mg/kg.
28. The method according to claim 23, in which HuLuc63 is
administered concurrently with the pharmaceutical composition
comprising one or more of said targeted agents, conventional
chemotherapeutic agents, hormonal therapy agents, supportive care
agents, or combinations thereof.
29. The method according to claim 28, in which said administration
elicits a complete response.
30. The method according to claim 28, in which said administration
elicits a very good partial response.
31. The method according to claim 28, in which said administration
elicits a partial response.
32. The method according to claim 28, in which said administration
elicits a minimal response.
33. A method of treating multiple myeloma in a subject, comprising
administering a therapeutically effective amount of HuLuc63, a
therapeutically effective amount of one or more targeted agents,
conventional chemotherapeutic agents, hormonal therapy agents,
supportive care agents, or combinations thereof, and a
pharmaceutically acceptable carrier, wherein said composition is
capable of being administered in a single or multiple dose regimen.
Description
1. CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims benefit under 35 U.S.C. .sctn.119(e)
to application Ser. Nos. 60/836,250 filed Aug. 7, 2006 and
60/856,144 filed Nov. 1, 2006, the contents of which are
incorporated herein by reference.
2. BACKGROUND
[0002] Multiple myeloma ("MM") represents a malignant proliferation
of plasma cells derived from a single clone. The terms multiple
myeloma and myeloma are used interchangeably to refer to the same
condition. The myeloma tumor, its products, and the host response
to it result in a number of organ dysfunctions and symptoms of bone
pain or fracture, renal failure, susceptibility to infection,
anemia, hypocalcemia, and occasionally clotting abnormalities,
neurologic symptoms and vascular manifestations of hyperviscosity.
See D. Longo, in Harrison's Principles of Internal Medicine 14th
Edition, p. 713 (McGraw-Hill, New York, 1998). No effective
long-term treatment currently exists for MM. It is a malignant
disease of plasma cells, manifested as hyperproteinemia, anemia,
renal dysfunction, bone lesions, and immunodeficiency. MM is
difficult to diagnose early because there may be no symptoms in the
early stage. The disease has a progressive course with a median
duration of survival of six months when no treatment is given.
Systemic chemotherapy is the main treatment, and the current median
of survival with chemotherapy is about three years, however fewer
than 5% live longer than 10 years (See Anderson, K. et al., Annual
Meeting Report 1999. Recent Advances in the Biology and Treatment
of Multiple Myeloma (1999)).
[0003] While multiple myeloma is considered to be a drug-sensitive
disease, almost all patients who initially respond to chemotherapy
eventually relapse (See Anderson, K. et al., Annual Meeting Report
1999. Recent Advances in the Biology and Treatment of Multiple
Myeloma (1999)). Since the introduction of melphalan and prednisone
therapy for MM, numerous multi-drug chemotherapies including Vinca
alkaloid, anthracycline, and nitrosourea-based treatments have been
tested (See Case, D C et al., (1977) Am. J. Med 63:897 903);
however, there has been little improvement in outcome over the past
three decades (See Case, D C et al., (1977) Am, J. Med 63:897 903;
Otsuki, T. et al, (2000) Cancer Res. 60:1). New methods of
treatment, such as combination therapies utilizing monoclonal
antibodies, therapeutic agents, and small molecule inhibitors of
cellular receptors and/or proteins implicated in MM, are
needed.
3. SUMMARY
[0004] Described herein are compositions and methods useful for
exploiting the anti-tumor properties of anti-CS1 antibodies.
Anti-CS1 antibodies that can be used in the methods and
compositions are described in U.S. Patent Publication Nos.
2005/0025763 and 2006/0024296, the contents of which are
incorporated herein by reference. The anti-CS1 antibodies target
CS1 (CD2-subsetl), which is also known as SLAMF7, CRACC, 19A,
APEX-1, and FOAP12 (Genbank Accession Number NM.sub.--021181.3).
CS1, is a glycoprotein that is highly expressed in bone marrow
samples from patients diagnosed with MM. In both in vitro and in
vivo studies, anti-CS1 antibodies exhibit significant anti-myeloma
activity (see, e.g., U.S. Patent Publication Nos. 2005/0025763 and
2006/0024296, the contents of which are incorporated herein by
reference). By way of example, but not limitation, the anti-CS1
antibody, HuLuc63 effectively mediates lysis of myeloma cells via
antibody dependent cellular cytotoxicity (ADCC) (see, e.g., U.S.
Patent Publication No. 2005/0025763, the content of which is
incorporated herein by reference). In a myeloma mouse tumor model,
treatment with HuLuc63 significantly reduced tumor mass by more
than 50% (see, e.g., U.S. Patent Publication No. 2005/0025763, the
content of which is incorporated herein by reference).
[0005] The present disclosure relates to compositions and methods
for treating patients diagnosed with Monoclonal Gammopathy of
Undetermined Significance (MGUS), smoldering myeloma, asymptomatic
MM, and symptomatic MM, ranging from newly diagnosed to late stage
relapsed/refractory. In particular, the methods relate to the
administration of a pharmaceutical composition comprising an
anti-CS1 antibody in combination with one or more therapeutic
agents. Anti-CS1 antibodies are typically administered
intravenously at doses ranging from 0.5 to 20 mg/kg, from once a
week to once a month.
[0006] One or more therapeutic agents, such as targeted agents,
conventional chemotherapy agents, hormonal therapy agents, and
supportive care agents and/or combinations thereof, can be
administered concurrently, prior to, or following administration of
an anti-CS1 antibody. The agents can be administered separately or
combined in a cocktail and administered together as a single
composition. The composition(s) can be administered by any means
known in the art.
[0007] In some embodiments, administration of the pharmaceutical
compositions described herein increases the sensitivity of multiple
myeloma cells to a therapeutic agent. For example, inclusion of an
anti-CS1 antibody, such as HuLuc63, enhances the activity of
therapeutic agents, such that lower doses can be used in the
compositions and methods described herein.
[0008] In some embodiments, administration of the pharmaceutical
compositions described herein elicits at least one of the
beneficial responses as defined by the European Group for Blood and
Marrow transplantation (EBMT). For example, administration of the
pharmaceutical compositions described herein can result in a
complete response, partial response, minimal response, no change,
or plateau.
4. BRIEF DESCRIPTION OF THE FIGURES
[0009] FIG. 1 depicts the anti-tumor activity of HuLuc63 alone and
in combination with dexamethasone in an in vivo mouse multiple
myeloma xenograft model;
[0010] FIG. 2 depicts the anti-tumor activity of HuLuc63 alone,
thalidomide alone, and HuLuc63 in combination with thalidomide in
an in vivo mouse multiple myeloma xenograft model;
[0011] FIG. 3 depicts the anti-tumor activity of HuLuc63 alone,
thalidomide/dexamethasone, and HuLuc63 in combination with
thalidomide and dexamethasone in an in vivo mouse multiple myeloma
xenograft model; and
[0012] FIG. 4 depicts the anti-tumor activity of HuLuc63 alone and
in combination with bevacizumab in an in vivo mouse multiple
myeloma xenograft model.
5. DETAILED DESCRIPTION
[0013] The compositions described herein combine anti-CS1
antibodies with one or more therapeutic agents at specific doses to
potentiate or complement the anti-myeloma activities of the other.
Examples of suitable anti-CS1 antibodies include, but are not
limited to, isolated antibodies that bind one or more of the three
epitope clusters identified on CS1 and monoclonal antibodies
produced by the hybridoma cell lines: Luc2, Luc3, Luc15, Luc22,
Luc23, Luc29, Luc32, Luc34, Luc35, Luc37, Luc38, Luc39, Luc56,
Luc60, Luc63, Luc69, LucX.1, LucX.2 or Luc90. These monoclonal
antibodies are named as the antibodies: Luc2, Luc3, Luc15, Luc22,
Luc23, Luc29, Luc32, Luc34, Luc35, Luc37, Luc38, Luc39, Luc56,
Luc60, Luc63, Luc69, LucX and Luc90, respectively, hereafter.
Humanized versions are denoted by the prefix "hu" (see, e.g., U.S.
Patent Publication Nos. 2005/0025763 and 2006/0024296, the contents
of which are incorporated herein by reference).
[0014] In some embodiments, suitable anti-CS1 antibodies include
isolated antibodies that bind one or more of the three epitope
clusters identified on CS1 (SEQ ID NO: 1, Table 1 below; see, e.g.,
U.S. Patent Publication No. 2006/0024296, the content of which is
incorporated herein by reference). As disclosed in U.S. Patent
Publication No. 2006/0024296 and shown below in Table 1, the CS1
antibody binding sites have been grouped into 3 epitope clusters:
[0015] (1) the epitope defined by Luc90, which binds to hu50/mu50
(SEQ ID NO: 2). This epitope covers from about amino acid residue
23 to about amino acid residue 151 of human CS1. This epitope is
resided within the domain 1 (V domain) of the extracellular domain.
This epitope is also recognized by Luc34, LucX (including LucX.1
and LucX.2) and Luc69. [0016] (2) the epitope defined by Luc38,
which binds to mu25/hu75 (SEQ ID NO: 3) and hu50/mu50 (SEQ ID NO:
81). This epitope likely covers from about amino acid residue 68 to
about amino acid residue 151 of human CS1. This epitope is also
recognized by Luc5. [0017] (3) the epitope defined by Luc 63, which
binds to mu75/hu25 (SEQ ID NO: 4). This epitope covers from about
amino acid residue 170 to about amino acid residue 227 of human
CS1. This epitope is resided within domain 2 (C2 domain) of human
CS1. This epitope is also recognized by Luc4, Luc12, Luc23, Luc29,
Luc32 and Luc37.
[0018] The methods and pharmaceutical compositions are addressed in
more detail below, but typically include at least one anti-CS1
antibody as described above. In some embodiments, the
pharmaceutical compositions include the anti-CS1 antibody HuLuc63.
HuLuc63 is a humanized recombinant monoclonal IgG1 antibody
directed to human CS1. The amino acid sequence for the heavy chain
variable region (SEQ ID NO: 5) and the light chain variable region
(SEQ ID NO: 6) for HuLuc63 is disclosed in U.S. Patent Publication
No. 2005/0025763, the content of which is incorporated herein by
reference, and in Table 1.
TABLE-US-00001 TABLE 1 SEQ ID NO: Amino Acid Sequence SEQ ID Met
Ala Gly Ser Pro Thr Cys Leu Thr Leu Ile Tyr Ile Leu Trp Gln Leu NO:
1 Thr Gly Ser Ala Ala Ser Gly Pro Val Lys Glu Leu Val Gly Ser Val
Gly Gly Ala Val Thr Phe Pro Leu Lys Ser Lys Val Lys Gln Val Asp Ser
Ile Val Trp Thr Phe Asn Thr Thr Pro Leu Val Thr Ile Gln Pro Glu Gly
Gly Thr Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg Val Asp Phe Pro
Asp Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys Lys Asn Asp Ser Gly
Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu Gln Gln Pro Ser Thr Gln
Glu Tyr Val Leu His Val Tyr Glu His Leu Ser Lys Pro Lys Val Thr Met
Gly Leu Gln Ser Asn Lys Asn Gly Thr Cys Val Thr Asn Leu Thr Cys Cys
Met Glu His Gly Glu Glu Asp Val Ile Tyr Thr Trp Lys Ala Leu Gly Gln
Ala Ala Asn Glu Ser His Asn Gly Ser Ile Leu Pro Ile Ser Trp Arg Trp
Gly Glu Ser Asp Met Thr Phe Ile Cys Val Ala Arg Asn Pro Val Ser Arg
Asn Phe Ser Ser Pro Ile Leu Ala Arg Lys Leu Cys Glu Gly Ala Ala Asp
Asp Pro Asp Ser Ser Met Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu
Leu Ser Leu Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu Arg
Gln Glu Glu Tyr Ile Glu Glu Lys Lys Arg Val Asp Ile Cys Arg Glu Thr
Pro Asn Ile Cys Pro His Ser Gly Glu Asn Thr Glu Tyr Asp Thr Ile Pro
His Thr Asn Arg Thr Ile Leu Lys Glu Asp Pro Ala Asn Thr Val Tyr Ser
Thr Val Glu Ile Pro Lys Lys Met Glu Asn Pro His Ser Leu Leu Thr Met
Pro Asp Thr Pro Arg Leu Phe Ala Tyr Glu Asn Val Ile SEQ ID Met Ala
Gly Ser Pro Thr Cys Leu Thr Leu Ile Tyr Ile Leu Trp Gln Leu NO: 2
Thr Gly Ser Ala Ala Ser Gly Pro Val Lys Glu Leu Val Gly Ser Val Gly
Gly Ala Val Thr Phe Pro Leu Lys Ser Lys Val Lys Gln Val Asp Ser Ile
Val Trp Thr Phe Asn Thr Thr Pro Leu Val Thr Ile Gln Pro Glu Gly Gly
Thr Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg Val Asp Phe Pro Asp
Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys Lys Asn Asp Ser Gly Ile
Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu Gln Gln Pro Ser Thr Gln Glu
Tyr Val Leu His Val Tyr Glu His Leu Ser Lys Pro Lys Val Thr Ile Asp
Arg Gln Ser Asn Lys Asn Gly Thr Cys Val Ile Asn Leu Thr Cys Ser Thr
Asp Gln Asp Gly Glu Asn Val Thr Tyr Ser Trp Lys Ala Val Gly Gln Gly
Asp Asn Gln Phe His Asp Gly Ala Thr Leu Ser Ile Ala Trp Arg Ser Gly
Glu Lys Asp Gln Ala Leu Thr Cys Met Ala Arg Asn Pro Val Ser Asn Ser
Phe Ser Thr Pro Val Phe Pro Gln Lys Leu Cys Glu Asp Ala Ala Thr Asp
Leu Thr Ser Leu Arg Gly SEQ ID Met Ala Arg Phe Ser Thr Tyr Ile Ile
Phe Thr Ser Val Leu Cys Gln Leu NO: 3 Thr Val Thr Ala Ala Ser Gly
Thr Leu Lys Lys Val Ala Gly Ala Leu Asp Gly Ser Val Thr Phe Thr Leu
Asn Ile Thr Glu Ile Lys Val Asp Tyr Val Val Trp Thr Phe Asn Thr Phe
Phe Leu Ala Met Val Lys Lys Asp Gly Gly Thr Ile Ile Val Thr Gln Asn
Arg Asn Arg Glu Arg Val Asp Phe Pro Asp Gly Gly Tyr Ser Leu Lys Leu
Ser Lys Leu Lys Lys Asn Asp Ser Gly Ile Tyr Tyr Val Gly Ile Tyr Ser
Ser Ser Leu Gln Gln Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu
His Leu Ser Lys Pro Lys Val Thr Met Gly Leu Gln Ser Asn Lys Asn Gly
Thr Cys Val Thr Asn Leu Thr Cys Cys Met Glu His Gly Glu Glu Asp Val
Ile Tyr Thr Trp Lys Ala Leu Gly Gln Ala Ala Asn Glu Ser His Asn Gly
Ser Ile Leu Pro Ile Ser Trp Arg Trp Gly Glu Ser Asp Met Thr Phe Ile
Cys Val Ala Arg Asn Pro Val Ser Arg Asn Phe Ser Ser Pro Ile Leu Ala
Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser Ser Met Val SEQ
ID Met Ala Arg Phe Ser Thr Tyr Ile Ile Phe Thr Ser Val Leu Cys Gln
Leu NO: 4 Thr Val Thr Ala Ala Ser Gly Thr Leu Lys Lys Val Ala Gly
Ala Leu Asp Gly Ser Val Thr Phe Thr Leu Asn Ile Thr Glu Ile Lys Val
Asp Tyr Val Val Trp Thr Phe Asn Thr Phe Phe Leu Ala Met Val Lys Lys
Asp Gly Val Thr Ser Gln Ser Ser Asn Lys Glu Arg Ile Val Phe Pro Asp
Gly Leu Tyr Ser Met Lys Leu Ser Gln Leu Lys Lys Asn Asp Ser Gly Ala
Tyr Arg Ala Glu Ile Tyr Ser Thr Ser Ser Gln Ala Ser Leu Ile Gln Glu
Tyr Val Leu His Val Tyr Lys His Leu Ser Arg Pro Lys Val Thr Ile Asp
Arg Gln Ser Asn Lys Asn Gly Thr Cys Val Ile Asn Leu Thr Cys Ser Thr
Asp Gln Asp Gly Glu Asn Val Thr Tyr Ser Trp Lys Ala Val Gly Gln Ala
Ala Asn Glu Ser His Asn Gly Ser Ile Leu Pro Ile Ser Trp Arg Trp Gly
Glu Ser Asp Met Thr Phe Ile Cys Val Ala Arg Asn Pro Val Ser Arg Asn
Phe Ser Ser Pro Ile Leu Ala Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp
Pro Asp Ser Ser Met Val SEQ ID Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Gln Pro Gly Gly Ser NO: 5 Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Asp Phe Ser Arg Tyr Trp Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Asp Ser Ser Thr Ile
Asn Tyr Ala Pro Ser Leu Lys Asp Lys Phe Ile Ile Ser Arg Asp Asn Ala
Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys Ala Arg Pro Asp Gly Asn Tyr Trp Tyr Phe Asp Val Trp
Gly Gln Gly Thr Leu Val Thr Val Ser Ser SEQ ID Asp Ile Gln Met Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp NO: 6 Arg Val Thr
Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Ile Ala Val Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile Tyr Tip Ala Ser Thr Arg
His Thr Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys
Gln Gln Tyr Ser Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu
Ile Lys
[0019] At some doses, additive effects are seen; at other doses,
synergistic effects are seen. In some embodiments, the synergistic
effect permits one or more therapeutic agents to be administered in
combination with one or more anti-CS1 antibodies at a reduced
dosage, while retaining efficacy. Given that the side effects
associated with the use of these agents are dose-dependent, use of
the compositions and methods described herein can reduce the
deleterious side effects observed in conventional and novel
treatment regimens used to treat MM when these agents are
administered at their recommended dosages.
[0020] In other embodiments, the synergistic effect permits one or
more therapeutic agents to be administered in combination with one
or more anti-CS1 antibodies at the approved dosage, but with
greater than the expected efficacy.
[0021] The compositions can be administered for the treatment of
Monoclonal Gammopathy of Undetermined Significance (MGUS),
smoldering myeloma, asymptomatic MM, and symptomatic MM, ranging
from newly diagnosed to late stage relapsed/refractory. Typically,
administration of the compositions results in a reduction in
M-protein in serum or urine such that a plateau, no change,
minimal, partial or complete response is observed as defined by the
European Group for Blood and Marrow transplantation (EBMT).
[0022] 5.2 Pharmaceutical Compositions
[0023] Provided herein are pharmaceutical compositions that are
beneficial in reducing tumor mass and/or regressing tumor growth,
in patients diagnosed with multiple myeloma. In addition, the
pharmaceutical compositions can be used to treat other diseases
characterized by the presence of monoclonal protein (M-protein,
paraprotein) in the serum or urine.
[0024] In some embodiments, the various components of the
compositions are provided separately. For example, an anti-CS1
antibody can be provided in a first pharmaceutical composition, and
a therapeutic agent provided in a second composition. When the
composition comprises two or more therapeutic agents, an anti-CS1
antibody can be provided in a first pharmaceutical composition, one
therapeutic agent can be provided in a second composition and the
other therapeutic agent can be provided in a third composition. In
other embodiments, an anti-CS1 antibody can be provided in one
pharmaceutical composition and the therapeutic agents can be
combined and provided in a second pharmaceutical composition. In
still other embodiments, one composition, comprising an anti-CS1
antibody and one or more therapeutic agents can be provided.
[0025] In typical embodiments, an anti-CS1 antibody is present in a
pharmaceutical composition at a concentration sufficient to permit
intravenous administration at 0.5 mg/kg to 20 mg/kg. In some
embodiments, the concentration of an anti-CS1 antibody suitable for
use in the compositions and methods described herein includes, but
is not limited to, at least about 0.5 mg/kg, at least about 0.75
mg/kg, at least about 1 mg/kg, at least about 2 mg/kg, at least
about 2.5 mg/kg, at least about 3 mg/kg, at least about 4 mg/kg, at
least about 5 mg/kg, at least about 6 mg/kg, at least about 7
mg/kg, at least about 8 mg/kg, at least about 9 mg/kg, at least
about 10 mg/kg, at least about 11 mg/kg, at least about 12 mg/kg,
at least about 13 mg/kg, at least about 14 mg/kg, at least about 15
mg/kg, at least about 16 mg/kg, at least about 17 mg/kg, at least
about 18 mg/kg, at least about 19 mg/kg, and at least about 20
mg/kg.
[0026] The anti-CS1 antibodies can be administered in single or
multiple dose regimens. Generally, an anti-CS1 antibody is
administered over a period of time from about 1 to about 24 hours,
but is typically administered over a period of about 1 to 2 hours.
Dosages can be repeated from about 1 to about 4 weeks or more, for
a total of 4 or more doses. Typically, dosages are repeated once
every week, once every other week, or once a month for a minimum of
4 doses to a maximum of 52 doses.
[0027] Determination of the effective dosage, total number of
doses, and length of treatment with an anti-CS1 antibody is well
within the capabilities of those skilled in the art, and can be
determined using a standard dose escalation study to identify the
maximum tolerated dose (MTh) (see, e.g., Richardson et al., 2002,
Blood, 100(9):3063-3067, the content of which is incorporated
herein by reference).
[0028] In some embodiments, one or more therapeutic agents are
administered in combination with an anti-CS1 antibody. The agents
can be administered concurrently, prior to, or following
administration of an anti-CS1 antibody.
[0029] In some embodiments, an anti-CS1 antibody is administered
prior to the administration of the therapeutic agents. For example,
the anti-CS1 antibody can be administered approximately 0 to 60
days prior to the administration of the therapeutic agents. In some
embodiments, an anti-CS1 antibody, such as HuLuc63, is administered
from about 30 minutes to about 1 hour prior to the administration
of the therapeutic agents, or from about 1 hour to about 2 hours
prior to the administration of the therapeutic agents, or from
about 2 hours to about 4 hours prior to the administration of the
therapeutic agents, or from about 4 hours to about 6 hours prior to
the administration of the therapeutic agents, or from about 6 hours
to about 8 hours prior to the administration of the therapeutic
agents, or from about 8 hours to about 16 hours prior to the
administration of the therapeutic agents, or from about 16 hours to
1 day prior to the administration of the therapeutic agents, or
from about 1 to 5 days prior to the administration of the
therapeutic agents, or from about 5 to 10 days prior to the
administration of the therapeutic agents, or from about 10 to 15
days prior to the administration of the therapeutic agents, or from
about 15 to 20 days prior to the administration of the therapeutic
agents, or from about 20 to 30 days prior to the administration of
the therapeutic agents, or from about 30 to 40 days prior to the
administration of the therapeutic agents, and from about 40 to 50
days prior to the administration of the therapeutic agents, or from
about 50 to 60 days prior to the administration of the therapeutic
agents.
[0030] In some embodiments, the anti-CS1 antibody is administered
concurrently with the administration of the therapeutic agents.
[0031] In some embodiments, an anti-CS1 antibody is administered
following the administration of the therapeutic agents. For
example, an anti-CS1 antibody, such as HuLuc63, can be administered
approximately 0 to 60 days after the administration of the
therapeutic agents. In some embodiments, HuLuc63 is administered
from about 30 minutes to about 1 hour following the administration
of the therapeutic agents, or from about 1 hour to about 2 hours
following the administration of the therapeutic agents, or from
about 2 hours to about 4 hours following the administration of the
therapeutic agents, or from about 4 hours to about 6 hours
following the administration of the therapeutic agents, or from
about 6 hours to about 8 hours following the administration of the
therapeutic agents, or from about 8 hours to about 16 hours
following the administration of the therapeutic agents, or from
about 16 hours to 1 day following the administration of the
therapeutic agents, or from about 1 to 5 days following the
administration of the therapeutic agents, or from about 5 to 10
days following the administration of the therapeutic agents, or
from about 10 to 15 days following the administration of the
therapeutic agents, or from about 15 to 20 days following the
administration of the therapeutic agents, or from about 20 to 30
days following the administration of the therapeutic agents, or
from about 30 to 40 days following the administration of the
therapeutic agents, and from about 40 to 50 days following the
administration of the therapeutic agents, or from about 50 to 60
days following the administration of the therapeutic agents.
[0032] Therapeutic agents that can be used in combination with the
anti-CS1 antibodies described herein include, but are not limited
to, targeted agents, conventional chemotherapy agents, hormonal
therapy agents, and supportive care agents. One or more therapeutic
agents from the different classes, e.g., targeted, conventional
chemotherapeutic, hormonal, and supportive care, and/or subclasses
can be combined in the compositions described herein. The various
classes described herein can be further divided into subclasses. By
way of example, targeted agents can be separated into a number of
different subclasses depending on their mechanism of action. As
will be apparent to those of skill in the art, the agents can have
more than one mechanism of action, and thus, could be classified
into one or more subclasses. For purposes of the compositions and
methods described herein, the following subclasses have been
identified: anti-angiogenic, inhibitors of growth factor signaling,
immunomodulators, inhibitors of protein synthesis, folding and/or
degradation, inhibitors of gene expression, pro-apoptotic agents,
agents that inhibit signal transduction and agents with "other"
mechanisms of action. Typically, the mechanism of action for agents
falling into the "other" subclass is unknown or poorly
characterized.
[0033] For example, in some embodiments, targeted agents, such as
bevacizumab, sutinib, sorafenib, 2-methoxyestradiol or 2ME2,
finasunate, PTK787, vandetanib, aflibercept, volociximab,
etaracizumab (MEDI-522), cilengitide, erlotinib, cetuximab,
panitumumab, gefitinib, trastuzumab, TKI258, CP-751,871, atacicept,
rituximab, alemtuzumab, aldesleukine, atlizumab, tocilizumab,
temsirolimus, everolimus, NPI-1387, MLNM3897, HCD122, SGN-40, HLL1,
huN901-DM1, atiprimod, natalizumab, bortezomib, carfilzomib,
NPI-0052, tanespimycin, saquinavir mesylate, ritonavir, nelfinavir
mesylate, indinavir sulfate, belinostat, LBH589, mapatumumab,
lexatumumab, AMG951, ABT-737, oblimersen, plitidepsin, SCIO-469,
P276-00, enzastaurin, tipifarnib, perifosine, imatinib, dasatinib,
lenalidomide, thalidomide, simvastatin, and celecoxib can be
combined with an anti-CS1 antibody, such as HuLuc63 and used to
treat MM patients.
[0034] By way of another example, conventional chemotherapy agents,
such as alklyating agents (e.g., oxaliplatin, carboplatin,
cisplatin, cyclophosphamide, melphalan, ifosfamide, uramustine,
chlorambucil, carmustine, mechloethamine, thiotepa, busulfan,
temozolomide, dacarbazine), anti-metabolic agents (e.g.,
gemcitabine, cytosine arabinoside, Ara-C, capecitabine, 5FU
(5-fluorouracil), azathioprine, mercaptopurine (6-MP),
6-thioguanine, aminopterin, pemetrexed, methotrexate), plant
alkaloid and terpenoids (e.g., docetaxel, paclitaxel, vincristine,
vinblastin, vinorelbine, vindesine, etoposide, VP-16, teniposide,
irinotecan, topotecan), anti-tumor antibiotics (e.g., dactinomycin,
doxorubicin, liposomal doxorubicin, daunorubicin, daunomycin,
epirubicin, mitoxantrone, adriamycin, bleomycin, plicamycin,
mitomycin C, caminomycin, esperamicins), and other agents (e.g.,
darinaparsin) can be combined with an anti-CS1 antibody, such as
HuLuc63 and used to treat MM.
[0035] By way of another example, hormonal agents such as
anastrozole, letrozole, goserelin, tamoxifen, dexamethasone,
prednisone, and prednisilone can be combined with an anti-CS1
antibody, such as HuLuc63 and used to treat MM.
[0036] By way of another example, supportive care agents such as
pamidronate, zoledonic acid, ibandronate, gallium nitrate,
denosumab, darbepotin alpha, epoetin alpha, eltrombopag, and
pegfilgrastim can be combined with an anti-CS1 antibody, such as
HuLuc63 and used to treat MM.
[0037] The therapeutic agents can be administered in any manner
found appropriate by a clinician and are typically provided in
generally accepted efficacious dose ranges, such as those described
in the Physician Desk Reference, 56th Ed. (2002), Publisher Medical
Economics, New Jersey. In other embodiments, a standard dose
escalation study can be performed to identify the maximum tolerated
dose (MTD) (see, e.g., Richardson, et al. 2002, Blood,
100(9):3063-3067, the content of which is incorporated herein by
reference).
[0038] In some embodiments, doses less than the generally accepted
efficacious dose of a therapeutic agent can be used. For example,
in various embodiments, the composition comprises a dosage that is
less than about 10% to 75% of the generally accepted efficacious
dose range. In some embodiments, at least about 10% or less of the
generally accepted efficacious dose range is used, at least about
15% or less, at least about 25%, at least about 30% or less, at
least about 40% or less, at least about 50% or less, at least about
60% or less, at least about 75% or less, and at least about
90%.
[0039] The therapeutic agents can be administered singly or
sequentially, or in a cocktail with other therapeutic agents, as
described below. The therapeutic agents can be administered orally,
intravenously, systemically by injection intramuscularly,
subcutaneously, intrathecally or intraperitoneally.
[0040] In some embodiments, the therapeutic agents provided in the
pharmaceutical composition(s) are selected from the group
consisting of dexamethasone, thalidomide, vincristine, carmustine
(BCNU), melphalan, cyclophosphamide, prednisone, doxorubicin,
cisplatin, etoposide, bortezomib, lenalidomide, ara-C, and/or
combinations thereof.
[0041] In certain embodiments, however, the pharmaceutical
composition does not comprise bortezomib and/or lenalidomide.
[0042] Accordingly, in some embodiments, two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of
dexamethasone.
[0043] In some embodiments, at least two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of
dexamethasone and thalidomide. In some embodiments, dexamethasone
and thalidomide are provided separately, such that a total of three
pharmaceutical compositions are provided: a first comprising an
anti-CS1 antibody such as HuLuc63, a second comprising
dexamethasone, and a third comprising thalidomide.
[0044] In some embodiments, at least two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of
vincristine, doxorubicin and dexamethasone (e.g., VAD). In some
embodiments, vincristine, doxorubicin and dexamethasone are
provided separately. Provided that the agents retain their
efficacy, compositions comprising other combinations of agents can
be prepared depending in part, on dosage, route of administration,
and whether the agents are provided in a solid, semi-solid or
liquid form. For example, a total of three compositions can be
made: a first comprising a therapeutically effective amount of an
anti-CS1 antibody such as HuLuc63, a second comprising
dexamethasone, and a third comprising vincristine and
doxorubicin.
[0045] In some embodiments, at least two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of doxorubicin
HCl liposome injection, vincristine and dexamethasone (e.g., DVd).
Provided that the agents retain their efficacy, compositions
comprising other combinations of agents can be prepared depending
in part, on dosage, route of administration, and whether the agents
are provided in a solid, semi-solid or liquid form. For example, a
total of three compositions can be made: a first comprising a
therapeutically effective amount of an anti-CS1 antibody such as
HuLuc63, a second comprising doxorubicin HCl liposome injection,
and a third comprising vincristine and dexamethasone.
[0046] In some embodiments, two pharmaceutical compositions are
provided: a first comprising a therapeutically effective amount of
an anti-CS1 antibody such as HuLuc63 and a second comprising a
therapeutically effective amount of cyclophosphamide.
[0047] In some embodiments, at least two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of
dexamethasone, thalidomide, cisplatin, doxorubicin,
cyclophosphamide and etoposide (e.g., DT-PACE). Provided that the
agents retain their efficacy, compositions comprising other
combinations can be prepared depending in part, on dosage, route of
administration, and whether the agents are provided in a solid,
semi-solid or liquid form. By way of example, dexamethasone and
thalidomide could be provided in one composition and cisplatin,
doxorubicin, cyclophosphamide and etoposide provided in another
composition.
[0048] In some embodiments, at least two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of
vincristine, doxorubicin, dexamethasone, and cyclophosphamide.
Provided that the agents retain their efficacy, compositions
comprising other combinations can be prepared depending in part, on
dosage, route of administration, and whether the agents are
provided in a solid, semi-solid or liquid form. By way of example,
vincristine, doxorubicin, and cyclophosphamide could be provided as
one composition and dexamethasone as a second.
[0049] In some embodiments, at least two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of doxorubicin
HCl liposome injection, vincristine, dexamethasone, and
thalidomide. Provided that the agents retain their efficacy,
compositions comprising other combinations can be prepared
depending in part, on dosage, route of administration, and whether
the agents are provided in a solid, semi-solid or liquid form. By
way of example, vincristine, dexamethasone, and thalidomide could
be provided as one composition and doxorubicin HCl liposome
injection as a second.
[0050] In some embodiments, at least two pharmaceutical
compositions are provided: a first comprising a therapeutically
effective amount of an anti-CS1 antibody such as HuLuc63 and a
second comprising a therapeutically effective amount of doxorubicin
HCl liposome injection and bortezomib. Provided that the agents
retain their efficacy, compositions comprising other combinations
can be prepared depending in part, on dosage, route of
administration, and whether the agents are provided in a solid,
semi-solid or liquid form. By way of example, doxorubicin HCl
liposome injection can be provided as one composition and
bortezomib as a second.
[0051] In some embodiments, agents with the same mechanism of
action as an anti-CS1 antibody, such as HuMax-Cd38 (Genmab) can be
provided in the pharmaceutical compositions described herein.
[0052] In some embodiments, the pharmaceutical compositions
comprise therapeutic agents with a mechanism of action that differs
from an anti-CS1 antibody. For example, targeted agents that
inhibit angiogenesis, including, but not limited to, bevacizumab,
sutinib, sorafenib, 2-methoxyestradiol or 2ME2, finasunate, PTK787,
vandetanib, aflibercept, volociximab, etaracizumab (MEDI-522),
cilengitide, can be used in the pharmaceutical compositions
described herein. In other embodiments, agents that inhibit growth
factor signaling, including, but not limited to, erlotinib,
cetuximab, panitumumab, gefitinib, trastuzumab, TKI258, CP-751,871,
atacicept, can be used in the pharmaceutical compositions described
herein. In other embodiments, immunomodulators including, but not
limited to, rituximab, alemtuzumab, aldesleukine, atlizumab,
tocilizumab, temsirolimus, everolimus, NPI-1387, MLNM3897, HCD122,
SGN-40, HLL1, huN901-DM1, atiprimod, natalizumab, can be used in
the pharmaceutical compositions described herein. In other
embodiments, agents that inhibit protein synthesis, folding or
degradation, including but not limited to, bortezomib, carfilzomib,
NPI-0052, tanespimycin, saquinavir mesylate, ritonavir, nelfinavir
mesylate, indinavir sulfate, can be used in the pharmaceutical
compositions described herein. In other embodiments, agents that
inhibit gene expression including but not limited to, belinostat,
LBH589, can be used in the pharmaceutical compositions described
herein. In other embodiments, pro-apoptotic agents including but
not limited to, mapatumumab, lexatumumab, AMG951, ABT-737,
oblimersen, plitidepsin, can be used in the pharmaceutical
compositions described herein. In other embodiments, agents that
inhibit signal transduction including but not limited to, SCIO-469,
P276-00, enzastaurin, tipifarnib, perifosine, imatinib, can be used
in the pharmaceutical compositions described herein. In other
embodiments, agents with other mechanisms of action including but
not limited to, dasatinib, lenalidomide, thalidomide, simvastatin,
and celecoxib can be used in the pharmaceutical compositions
described herein.
[0053] The pharmaceutical compositions can exist as a solid,
semi-solid, or liquid (e.g., suspensions or aerosols) dosage form.
Typically, the compositions are administered in unit dosage forms
suitable for single administration of precise dosage amounts. For
example, an anti-CS1 antibody can be packaged in dosages ranging
from about 1 to 1000 mg. In some embodiments, an anti-CS1 antibody
is packaged in a dosage at least about 1 mg, at least about 10 mg,
at least about 20 mg, at least about 50 mg, at least about 100 mg,
at least about 200 mg, at least about 300 mg, at least about 400
mg, at least about 500 mg, at least about 750 mg, at least about
1000 mg.
[0054] The compositions can also include, depending on the
formulation desired, pharmaceutically-acceptable, nontoxic carriers
or diluents, which are defined as vehicles commonly used to
formulate pharmaceutical compositions for animal or human
administration. The diluent is selected so as not to affect the
biological activity of the combination. Examples of such diluents
are distilled water, physiological saline, Ringer's solution,
dextrose solution, and Hank's solution.
[0055] In addition, the pharmaceutical composition or formulation
can also include other carriers, adjuvants, or nontoxic,
non-therapeutic, nonimmunogenic stabilizers and the like. Effective
amounts of such diluent or carrier will be those amounts that are
effective to obtain a pharmaceutically acceptable formulation in
terms of solubility of components, or biological activity.
[0056] 5.3 Use of the Pharmaceutical Compositions to Treat Multiple
Myeloma
[0057] The pharmaceutical compositions described herein find use in
treating MM.
[0058] Typically, the compositions can be used to treat Monoclonal
Gammopathy of Undetermined Significance (MGUS), smoldering myeloma,
asymptomatic MM, and symptomatic MM, ranging from newly diagnosed
to late stage relapsed/refractory.
[0059] The compositions can be combined with other treatment
strategies, i.e., autologous stem cell transplantation and
allogeneic effector cell transplantation, to develop an effective
treatment strategy based on the stage of myeloma being treated
(see, e.g., Multiple Myeloma Research Foundation, Multiple Myeloma:
Stem Cell Transplantation 1-30 (2004); U.S. Pat. Nos. 6,143,292,
and 5,928,639, Igarashi, et al. Blood 2004, 104(1): 170-177,
Maloney, et al. 2003, Blood, 102(9): 3447-3454, Badros, et al.
2002, J Clin Oncol., 20:1295-1303, Tricot, et al. 1996, Blood,
87(3):1196-1198; the contents of which are incorporated herein by
reference).
[0060] The staging system most widely used since 1975 has been the
Durie-Salmon system, in which the clinical stage of disease (Stage
I, II, or III) is based on four measurements (see, e.g., Durie and
Salmon, 1975, Cancer, 36:842-854). These four measurements are: (1)
levels of monoclonal (M) protein (also known as paraprotein) in the
serum and/or the urine; (2) the number of lytic bone lesions; (3)
hemoglobin values; and, (4) serum calcium levels. These three
stages can be further divided according to renal function,
classified as A (relatively normal renal function, serum creatinine
value <2.0 mg/dL) and B (abnormal renal function, creatinine
value .gtoreq.2.0 mg/dL). A new, simpler alternative is the
International Staging System (ISS) (see, e.g., Greipp et al., 2003,
"Development of an international prognostic index (IPI) for
myeloma: report of the international myeloma working group", The
Hematology). The ISS is based on the assessment of two blood test
results, beta.sub.2-microglobulin (.beta..sub.2-M) and albumin,
which separates patients into three prognostic groups irrespective
of type of therapy.
[0061] Administration of the pharmaceutical compositions at
selected dosage ranges and routes typically elicits a beneficial
response as defined by the European Group for Blood and Marrow
transplantation (EBMT). Table 2 lists the EBMT criteria for
response.
TABLE-US-00002 TABLE 2 EBMT/IBMTR/ABMTR.sup.1 Criteria for Response
Complete Response No M-protein detected in serum or urine by
immunofixation for a minimum of 6 weeks and fewer than 5% plasma
cells in bone marrow Partial Response >50% reduction in serum
M-protein level and/or 90% reduction in urine free light chain
excretion or reduction to <200 mg/24 hrs for 6 weeks.sup.2
Minimal Response 25-49% reduction in serum M-protein level and/or
50-89% reduction in urine free light chain excretion which still
exceeds 200 mg/24 hrs for 6 weeks.sup.3 No Change Not meeting the
criteria or either minimal response or progressive disease Plateau
No evidence of continuing myeloma-related organ or tissue damage,
<25% change in M- protein levels and light chain excretion for 3
months Progressive Disease Myeloma-related organ or tissue damage
continuing despite therapy or its reappearance in plateau phase,
>25% increase in serum M- protein level (>5 g/L) and/or
>25% increase in urine M-protein level (>200 mg/24 hrs)
and/or >25% increase in bone marrow plasma cells (at least 10%
in absolute terms).sup.2 Relapse Reappearance of disease in
patients previously in complete response, including detection of
paraprotein by immunofixation .sup.1EBMT: European Group for Blood
and Marrow transplantation; IBMTR: International Bone Marrow
Transplant Registry; ABMTR: Autologous Blood and Marrow Transplant
Registry. .sup.2For patients with non-secretory myeloma only,
reduction of plasma cells in the bone marrow by >50% of initial
number (partial response) or 25-49% of initial number (minimal
response) is required. .sup.3In non-secretory myeloma, bone marrow
plasma cells should increase by >25% and at least 10% in
absolute terms; MRI examination may be helpful in selected
patients.
[0062] Additional criteria that can be used to measure the outcome
of a treatment include "near complete response" and "very good
partial response". A "near complete response" is defined as the
criteria for a "complete response" (CR), but with a positive
immunofixation test. A "very good partial response" is defined as a
greater than 90% decrease in M protein (see, e.g., Multiple Myeloma
Research Foundation, Multiple Myeloma: Treatment Overview 9
(2005)).
[0063] The degree to which administration of the compositions
elicits a response in an individual clinically manifesting at least
one symptom associated with MM, depends in part, on the severity of
disease, e.g., Stage I, II, or III, and in part, on whether the
patient is newly diagnosed or has late stage refractory MM. Thus,
in some embodiments, administration of the pharmaceutical
composition elicits a complete response.
[0064] In other embodiments, administration of the pharmaceutical
composition elicits a very good partial response or a partial
response.
[0065] In other embodiments, administration of the pharmaceutical
composition elicits a minimal response.
[0066] In other embodiments, administration of the pharmaceutical
composition prevents the disease from progressing, resulting in a
response classified as "no change" or "plateau" by the EBMT.
[0067] Routes of administration and dosage ranges for compositions
comprising an anti-CS1 antibody such as HuLuc63 and one or more
therapeutic agents for treating individuals diagnosed with MM, can
be determined using art-standard techniques, such as a standard
dose escalation study to identify the MTD (see, e.g., Richardson,
et al. 2002, Blood, 100(9):3063-3067, the content of which is
incorporated herein by reference).
[0068] Typically, anti-CS1 antibodies are administered
intravenously. Administration of the other therapeutic agents
described herein can be by any means known in the art. Such means
include oral, rectal, nasal, topical (including buccal and
sublingual) or parenteral (including subcutaneous, intramuscular,
intravenous and intradermal) administration and will depend in
part, on the available dosage form. For example, therapeutic agents
that are available in a pill or capsule format typically are
administered orally. However, oral administration generally
requires administration of a higher dose than does intravenous
administration. Determination of the actual route of administration
that is best in a particular case is well within the capabilities
of those skilled in the art, and in part, will depend on the dose
needed versus the number of times per month administration is
required.
[0069] Factors affecting the selected dosage of an anti-CS1
antibody and the therapeutic agents used in the compositions and
methods described herein, include, but are not limited to, the type
of agent, the age, weight, and clinical condition of the recipient
patient, and the experience and judgment of the clinician or
practitioner administering the therapy. Generally, the selected
dosage should be sufficient to result in no change, but preferably
results in at least a minimal change. An effective amount of a
pharmaceutical agent is that which provides an objectively
identifiable response, e.g., plateau, no change, minimal, partial,
or complete, as noted by the clinician or other qualified observer,
and as defined by the EBMT.
[0070] Generally, an anti-CS1 antibody is administered as a
separate composition from the composition(s) comprising the
therapeutic agents. As discussed above, the therapeutic agents can
each be administered as a separate composition, or combined in a
cocktail and administered as a single combined composition. In some
embodiments, the compositions comprising an anti-CS1 antibody and
one or more therapeutic agents are administered concurrently. In
other embodiments, an anti-CS1 antibody can be administered prior
to the administration of composition(s) comprising the therapeutic
agent(s). In yet other embodiments, an anti-CS1 antibody is
administered following the administration of composition(s)
comprising the therapeutic agent(s).
[0071] In those embodiments in which the an anti-CS1 antibody is
administered prior to or following the administration of the
therapeutic agents, determination of the duration between the
administration of an anti-CS1 antibody and administration of the
agents is well within the capabilities of those skilled in the art,
and in part, will depend on the dose needed versus the number of
times per month administration is required.
[0072] Doses of anti-CS1 antibodies used in the methods described
herein typically range between 0.5 mg/kg to 20 mg/kg. Optimal doses
for the therapeutic agents are the generally accepted efficacious
doses, such as those described in the Physician Desk Reference,
56th Ed. (2002), Publisher Medical Economics, New Jersey. Optimal
doses for agents not described in the Physician Desk Reference can
be determined using a standard dose escalation study to identify
the MTD (see, e.g., Richardson, et al. 2002, Blood,
100(9):3063-3067, the contents of which are incorporated herein by
reference).
[0073] In some embodiments, an anti-CS1 antibody is present in a
pharmaceutical composition at a concentration, or in a
weight/volume percentage, or in a weight amount, suitable for
intravenous administration at a dosage rate at least about 0.5
mg/kg, at least about 0.75 mg/kg, at least about 1 mg/kg, at least
about 2 mg/kg, at least about 2.5 mg/kg, at least about 3 mg/kg, at
least about 4 mg/kg, at least about 5 mg/kg, at least about 6
mg/kg, at least about 7 mg/kg, at least about 8 mg/kg, at least
about 9 mg/kg, at least about 10 mg/kg, at least about 11 mg/kg, at
least about 12 mg/kg, at least about 13 mg/kg, at least about 14
mg/kg, at least about 15 mg/kg, at least about 16 mg/kg, at least
about 17 mg/kg, at least about 18 mg/kg, at least about 19 mg/kg,
and at least about 20 mg/kg.
6. EXAMPLES
Example 1
HuLuc63 in Combination with Dexamethasone
[0074] Dexamethasone (Dex) is a corticosteroid that is used
extensively for the treatment of MM in multiple lines of therapy.
Dex has been used as monotherapy or in combination with a variety
of agents, including lenalidomide, thalidomide, velcade or as part
of the VAD (vincristine, doxorubicin, dexamethasone) regimen or the
DVd liposomal doxorubicin, vincristine, short-schedule
dexamethasone regimen. The mechanism of action of Dex involves
direct induction of apoptosis of myeloma cells by activation of
caspases (Chauhan et al., 1997, Oncogene 15:837-843; Chauhan et
al., 1997, J. Biol. Chem. 272, 29995-29997; Chauhan et al., 2001,
J. Biol. Chem., 276: 24453-24456).
[0075] The effect of HuLuc63 and Dex treatment on the expression of
CS1 in MM cell lines and mouse xenograft tumors was examined by
flow cytometry and immunohistochemistry respectively.
[0076] In vivo Xenograft Mouse Model: Methods and Results
[0077] Six- to eight-week old female IcrTac:ICR-Prkdcscid mice
obtained from Taconic Farms (Germantown, N.Y.) were inoculated with
1.times.10.sup.7 OPM2 or L363 cells (German Collection of
Microorganisms and Cell Cultures, Braunschweig, Germany) into the
lower right flank. Caliper measurements were performed twice weekly
to calculate tumor volume using the following formula:
L.times.W.times.H/2, where L (length) is the longest side of the
tumor in the plane of the animal's back, W (width) is the longest
measurement perpendicular to the length and in the same plane and H
(height) is taken at the highest point perpendicular to the back of
the animal. When tumors reached an average size of about 100
mm.sup.3, animals were randomized into 3 groups of 8-10 mice each
and were treated with 1 or 10 mg/kg of HuLuc63 or isotype control
antibody administered intraperitoneally twice a week for a maximum
of 6 doses.
[0078] Dex was administered intraperitoneally at a dose of 10 mg/kg
twice a week for a maximum of 6 doses.
[0079] Tumor growth was monitored for a period of 1-2 months.
Animal work was carried out under NIH guidelines ("Guide for the
Care and Use of Laboratory Animals") using protocols approved by
IACUC at PDL BioPharma.
[0080] CS1 protein expression was examined on the OPM2 multiple
myeloma cell line. No significant change in CS1 expression was
observed pre- or post-treatment with HuLuc63, Dex, or with both
agents. The combination of HuLuc63 with Dex was tested for
anti-myeloma activity in vivo. OPM2 tumor-bearing mice were treated
with sub-optimal doses of HuLuc63 (1 mg/kg), or isotype control
antibody twice weekly for three weeks. Dex was given twice a week
at 10 mg/kg to mice receiving either isotype control antibody or
HuLuc63. The results showed significant anti-tumor activity of
HuLuc63 alone and in combination with Dex (see, e.g., FIG. 1). Mice
in the combination treatment group exhibited significantly smaller
tumors than in the HuLuc63 monotherapy group (see, e.g., FIG.
1).
Example 2
HuLuc63 in Combination with Thalidomide
[0081] Thalidomide (Thal) is an immunomodulatory drug that is
currently approved for the treatment of MM in combination with Dex.
The mechanism of action of Thal is not completely understood, but
involves inhibition of angiogenesis, inhibition of growth and
survival of stromal cells and tumor cells in the bone marrow, and
altering the production of factors that influence the survival of
myeloma cells (such as IL-6, IL-10, IL-4, IL-5, IL-12, IL-8,
TNF-alpha).
[0082] The effect of HuLuc63 and Thal treatment on the expression
of CS1 in MM cell lines and mouse xenograft tumors was examined by
flow cytometry and immunohistochemistry respectively, as described
above.
[0083] CS1 protein expression was examined on the L363 multiple
myeloma cell line. No significant change in CS1 expression was
observed pre- or post-treatment with HuLuc63, Thal or with both
agents. The combination of HuLuc63 with Thal was tested for
anti-myeloma activity in vivo. L363 tumor-bearing mice were treated
with HuLuc63, or isotype control antibody twice weekly for three
weeks at 10 mg/kg. Thal was given 5 days a week at 50 mg/kg for a
maximum of 15 doses to mice receiving either isotype control
antibody or HuLuc63. Significant anti-tumor activity was observed
with HuLuc63 alone, Thal alone and HuLuc63 in combination with Thal
(see, e.g., FIG. 2). Mice in the combination treatment group
exhibited smaller tumors compared to the HuLuc63 and Thal
monotherapy groups (see, e.g., FIG. 2).
Example 3
HuLuc63 in Combination with Thalidomide/Dexamethasone
[0084] Thal/Dex combination therapy is currently approved for
front-line treatment of MM patients. The combination of HuLuc63
with Thal/Dex was tested for anti-myeloma activity in vivo as
described above. The effect of Thal/Dex treatment on the expression
of CS1 in MM cell lines and mouse xenograft tumors was examined by
immunohistochemistry as described above.
[0085] L363 tumor-bearing mice were treated with HuLuc63, or
isotype control antibody twice weekly for three weeks at 10 mg/kg.
Thal was given 5 days a week at 50 mg/kg for a maximum of 15 doses
and Dex was given twice a week at 10 mg/kg for a maximum of 6 doses
to mice receiving either isotype control antibody or HuLuc63. The
results showed significant anti-tumor activity of HuLuc63 alone,
Thal/Dex alone and HuLuc63 in combination with Thal/Dex (see e.g.,
FIG. 3). Mice in the HuLuc63/Thal/Dex combination treatment group
exhibited the smallest tumors, on average exhibiting a decrease of
50-70% in tumor size compared to HuLuc63 monotherapy or Thal/Dex
therapy (see e.g., FIG. 3).
Example 4
HuLuc63 in Combination with Bevacizumab
[0086] Bevacizumab is a monoclonal antibody that targets the
endothelial growth factor VEGF. It works by inhibiting VEGF,
resulting in the inhibition of new blood vessel formation in
tumors. Bevacizumab is currently approved for the treatment of
solid tumors, including metastatic colorectal cancer. Bevacizumab
is currently not approved for the use in the treatment of MM.
[0087] To determine if an anti-angiogenic drug can enhance the
antitumor effects of HuLuc63, a combination of HuLuc63 with
Bevacizumab was tested for anti-myeloma activity in vivo, as
described above. L363 tumor-bearing mice were treated with HuLuc63,
or isotype control antibody twice weekly for three weeks at 10
mg/kg. Bevacizumab was given twice a week at 0.5 mg/kg for a
maximum of 6 doses to mice receiving either isotype control
antibody or HuLuc63. Significant anti-tumor activity of HuLuc63
alone and in combination with Bevacizumab was observed (see e.g.,
FIG. 4). Bevacizumab alone did not exhibit significant anti-tumor
activity. However, mice receiving the combination of HuLuc63 with
Bevacizumab exhibited significantly smaller tumors compared to the
HuLuc63 monotherapy group, indicating that HuLuc63 and Bevacizumab
may act synergistically to cause an anti-myeloma effect.
Example 5
Treatment of Patients Diagnosed with Multiple Myeloma
[0088] Proposed multi-center, open-label, multi-dose, dose
escalation studies will be used to evaluate the combination of
HuLuc63 and one or more therapeutic agents in patients with
multiple myeloma after 1.sup.st, 2.sup.nd, or 3.sup.rd relapse.
HuLuc63 will be given by intravenous injection (IV) at up to five
dose levels ranging from 2.5 mg/kg to 20 mg/kg in combination with
one or more therapeutic agents. Patients will receive HuLuc63 once
a week, with each dose infused over 1 hour, once every 10 days or
once every two weeks, for a minimum of 4 doses and a maximum of 52
doses.
[0089] Drug combinations suitable for administration with HuLuc63
include, but are not limited to: [0090] 1) Melphalan+prednisone.
Melphalan will be given in 28 day cycles, at 8 mg/m.sup.2/d on days
1 to 4, for a maximum number of 11 cycles. Prednisone will be given
will be given in 28 day cycles, at 60 mg/m.sup.2/d on days 1 to 4,
for a maximum number of 11 cycles; [0091] 2)
Melphalan+prednisone+thalidomide. Melphalan will be given in 28 day
cycles, at 8 mg/m.sup.2/d on days 1 to 4, for a maximum number of
11 cycles. Prednisone will be given will be given in 28 day cycles,
at 60 mg/m.sup.2/d on days 1 to 4, for a maximum number of 11
cycles. Thalidomide will be given in 28 day cycles, at 200 mg/d,
for a maximum number of 11 cycles; [0092] 3)
Melphalan+prednisone+lenalidomide. Melphalan will be given in 28 to
42 day cycles, at 0.18 to 0.25 mg/kg for 4 days every 4 to 6 weeks,
for a maximum number of 9 cycles. Prednisone will be given will be
given in 28 to 42 day cycles, at 2 mg/kg for 4 days every 4 to 6
weeks, for a maximum number of 9 cycles, Lenalidomide will be given
in 28 to 42 day cycles, at 5 to 10 mg/day, days 1 to 21 every 4 to
6 weeks, for a maximum number of 9 cycles; [0093] 4)
Melphalan+prednisone+bortezomib. Melphalan will be given in 42 day
cycles, at 9 mg/m.sup.2/day on days 1 to 4, for a maximum number of
4 cycles. Prednisone will be given will be given in 42 day cycles,
60 mg/m.sup.2/d on days 1 to 4, for a maximum number of 4 cycles.
Bortezomib will be given in 42 day cycles, at 1.3 mg/m.sup.2/day on
days 1, 4, 8, 11, 22, 25, 29 and 32, for a maximum number of 4
cycles; [0094] 5) Thalidomide+dexamethasone. Thalidomide will be
given in 28 day cycles, at 200 mg/d, for a maximum number of 4 to
12 cycles. Dexamethasone will be given at 40 mg/day on days 1 to 4,
9 to 12, and 17 to 20, for a maximum number of 4 to 12 cycles;
[0095] 6) Cytoxan+prednisone. Cytoxan will be given in 28 day
cycles, at 500 mg, once weekly, for a maximum number of 6 cycles.
Prednisone will be given in 28 day cycles, at 100 mg/day, every
other day, for a maximum number of 6 cycles; [0096] 7)
Cytoxan+prednisone+thalidomide. Cytoxan will be given in 28 day
cycles, at 50 mg twice per day for 21 days, for a maximum number of
7 cycles. Prednisone will be given in 28 day cycles, at 100 mg/day,
every other day, for a maximum number of 7 cycles. Thalidomide will
be given in 28 day cycles, at 200 mg/d, for a maximum number of 7
cycles; [0097] 8) Cytoxan+prednisone+lenalidomide. Cytoxan will be
given in 28 clay cycles, at 300 to 700 mg on days 1 and 8, for a
maximum number of 4 to 7 cycles. Prednisone will be given in 28 day
cycles, at 50 mg/day every other day, for a maximum number of 4 to
7 cycles. Lenalidomide will be given in 28 day cycles, at 25 mg/day
on days 1 to 21, for a maximum number of 4 to 7 cycles; [0098] 9)
Cytoxan+dexamethasone+lenalidomide. Cytoxan will be given in 28 day
cycles, at 300 to 700 mg on days 1 and 8, for a maximum number of 4
to 7 cycles. Dexamethasone will be given in 28 day cycles, at 40 mg
on days 1 to 4, 9 to 12 and 17 to 20, for a maximum number of 4 to
7 cycles. Lenalidomide will be given in 28 day cycles, at 25 mg/day
on days 1 to 21, for a maximum number of 4 to 7 cycles; [0099] 10)
Cytoxan+prednisone+bortezomib. Cytoxan will be given in 28 day
cycles, at 300 mg on days 1, 8, 15, and 22, for a maximum number of
4 to 7 cycles. Prednisone will be given in 28 day cycles, at 100 mg
every other day, for a maximum number of 4 to 7 cycles. Bortezomib
will be given in 28 day cycles, at 1.5 mg/m.sup.2/day, days 1, 8,
and 15, for a maximum number of 4 to 7 cycles; [0100] 11)
Bevacizumab. Bevacizumab will be given in 14 day cycles, at 5
mg/kg, every 2 weeks, for a maximum number of 6 to 26 cycles;
[0101] 12) Tanespimycin+bortezomib. Tanespimycin will be given in
21 day cycles, at 100 to 340 mg/m.sup.2, twice weekly for 2 weeks,
for a maximum of 4 to 8 cycles. Bortezomib will be given in 21 day
cycles, at 0.7 to 1.3 mg/m.sup.2, twice weekly for 2 weeks, for a
maximum number of 4 to 8 cycles; [0102] 13) Doxil+bortezomib. Doxil
will be given in 21 day cycles, at 30 mg/m.sup.2/day, at day 4, for
a maximum number of 4 to 8 cycles. Bortezomib will be given in 21
day cycles, at 1.3 mg/m.sup.2, days 1, 4, 8 and 11, for a maximum
number of 4 to 8 cycles; and, [0103] 14) Dexamethasone.
Dexamethasone can be given in 28 day cycles, at 40 mg/day, at days
1 to 4, 9 to 12, and 17 to 20, for a maximum number of 4 to 12
cycles, or alternately, in 28 day cycles, at 40 mg/day for 4 days,
in 28 day cycles.
[0104] The above drug combinations can be given orally or by
IV.
[0105] After 8-12 weeks of therapy, EBMT criteria will be assessed.
If a patient has progressive disease, HuLuc63 will be discontinued
and the other drug combinations may be withdrawn or continued at
the discretion of the site investigator. If the patient has
responded or has stable disease at Week 8-12, dosing with HuLuc63
and the drug combination will continue so that a maximum of 52
weeks of treatment are completed or disease progression occurs.
[0106] Approximately 15 to 30 patients in 5 cohorts will be
enrolled for each drug combination. Each cohort will begin with 3
patients. If no dose-limiting toxicity (DLT) is noted within the
first 4 weeks of treatment in any patient, enrollment will begin in
the next higher cohort. If one patient has a DLT, 3 additional
patients will be enrolled in the cohort. If no other patient in the
cohort has a DLT, escalation to the next cohort may proceed. If a
second patient in a cohort has a DLT, the maximum tolerated dose
(MTD) has been reached.
[0107] A dose-limiting toxicity (DLT) is defined using the National
Cancer Center Institute Common Toxicity Criteria Version 3.0 (NCI
CTCAE v3.0) as a grade 4 hematologic toxicity or
hyperbilirubinemia, or a grade 3 toxicity in any other system
considered related to HuLuc63 or the combination of HuLuc63 and any
of the drugs listed in Table 1. For dose escalation to the next
cohort, 3 assessable patients must complete their first 4 weeks of
dosing. If a DLT occurs, an additional three assessable patients
will be accrued. Patients will be monitored for safety by assessing
adverse events categorized by NCI CTCAE v3.0 and patients will be
monitored for clinical activity using EBMT. The maximally tolerated
dose (MTD) is defined as the highest dose studied for which the
incidence of DLTs is <33%. The highest tolerated dose will be
HuLuc63 at 20 mg/kg combined with the drugs at the doses listed
above if no dose limiting toxicities are observed.
[0108] All publications, patents, patent applications and other
documents cited in this application are hereby incorporated by
reference in their entireties for all purposes to the same extent
as if each individual publication, patent, patent application or
other document were individually indicated to be incorporated by
reference for all purposes.
[0109] While various specific embodiments have been illustrated and
described, it will be appreciated that various changes can be made
without departing from the spirit and scope of the invention(s).
Sequence CWU 1
1
61335PRTHomo sapiens 1Met Ala Gly Ser Pro Thr Cys Leu Thr Leu Ile
Tyr Ile Leu Trp Gln1 5 10 15Leu Thr Gly Ser Ala Ala Ser Gly Pro Val
Lys Glu Leu Val Gly Ser 20 25 30Val Gly Gly Ala Val Thr Phe Pro Leu
Lys Ser Lys Val Lys Gln Val 35 40 45Asp Ser Ile Val Trp Thr Phe Asn
Thr Thr Pro Leu Val Thr Ile Gln 50 55 60Pro Glu Gly Gly Thr Ile Ile
Val Thr Gln Asn Arg Asn Arg Glu Arg65 70 75 80Val Asp Phe Pro Asp
Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys 85 90 95Lys Asn Asp Ser
Gly Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu 100 105 110Gln Gln
Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu His Leu 115 120
125Ser Lys Pro Lys Val Thr Met Gly Leu Gln Ser Asn Lys Asn Gly Thr
130 135 140Cys Val Thr Asn Leu Thr Cys Cys Met Glu His Gly Glu Glu
Asp Val145 150 155 160Ile Tyr Thr Trp Lys Ala Leu Gly Gln Ala Ala
Asn Glu Ser His Asn 165 170 175Gly Ser Ile Leu Pro Ile Ser Trp Arg
Trp Gly Glu Ser Asp Met Thr 180 185 190Phe Ile Cys Val Ala Arg Asn
Pro Val Ser Arg Asn Phe Ser Ser Pro 195 200 205Ile Leu Ala Arg Lys
Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser 210 215 220Ser Met Val
Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu225 230 235
240Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu Arg Gln Glu
245 250 255Glu Tyr Ile Glu Glu Lys Lys Arg Val Asp Ile Cys Arg Glu
Thr Pro 260 265 270Asn Ile Cys Pro His Ser Gly Glu Asn Thr Glu Tyr
Asp Thr Ile Pro 275 280 285His Thr Asn Arg Thr Ile Leu Lys Glu Asp
Pro Ala Asn Thr Val Tyr 290 295 300Ser Thr Val Glu Ile Pro Lys Lys
Met Glu Asn Pro His Ser Leu Leu305 310 315 320Thr Met Pro Asp Thr
Pro Arg Leu Phe Ala Tyr Glu Asn Val Ile 325 330
3352227PRTArtificialhu50/mu50 2Met Ala Gly Ser Pro Thr Cys Leu Thr
Leu Ile Tyr Ile Leu Trp Gln1 5 10 15Leu Thr Gly Ser Ala Ala Ser Gly
Pro Val Lys Glu Leu Val Gly Ser 20 25 30Val Gly Gly Ala Val Thr Phe
Pro Leu Lys Ser Lys Val Lys Gln Val 35 40 45Asp Ser Ile Val Trp Thr
Phe Asn Thr Thr Pro Leu Val Thr Ile Gln 50 55 60Pro Glu Gly Gly Thr
Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg65 70 75 80Val Asp Phe
Pro Asp Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys 85 90 95Lys Asn
Asp Ser Gly Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu 100 105
110Gln Gln Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu His Leu
115 120 125Ser Lys Pro Lys Val Thr Ile Asp Arg Gln Ser Asn Lys Asn
Gly Thr 130 135 140Cys Val Ile Asn Leu Thr Cys Ser Thr Asp Gln Asp
Gly Glu Asn Val145 150 155 160Thr Tyr Ser Trp Lys Ala Val Gly Gln
Gly Asp Asn Gln Phe His Asp 165 170 175Gly Ala Thr Leu Ser Ile Ala
Trp Arg Ser Gly Glu Lys Asp Gln Ala 180 185 190Leu Thr Cys Met Ala
Arg Asn Pro Val Ser Asn Ser Phe Ser Thr Pro 195 200 205Val Phe Pro
Gln Lys Leu Cys Glu Asp Ala Ala Thr Asp Leu Thr Ser 210 215 220Leu
Arg Gly2253227PRTArtificialmu25/hu75 3Met Ala Arg Phe Ser Thr Tyr
Ile Ile Phe Thr Ser Val Leu Cys Gln1 5 10 15Leu Thr Val Thr Ala Ala
Ser Gly Thr Leu Lys Lys Val Ala Gly Ala 20 25 30Leu Asp Gly Ser Val
Thr Phe Thr Leu Asn Ile Thr Glu Ile Lys Val 35 40 45Asp Tyr Val Val
Trp Thr Phe Asn Thr Phe Phe Leu Ala Met Val Lys 50 55 60Lys Asp Gly
Gly Thr Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg65 70 75 80Val
Asp Phe Pro Asp Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys 85 90
95Lys Asn Asp Ser Gly Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu
100 105 110Gln Gln Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu
His Leu 115 120 125Ser Lys Pro Lys Val Thr Met Gly Leu Gln Ser Asn
Lys Asn Gly Thr 130 135 140Cys Val Thr Asn Leu Thr Cys Cys Met Glu
His Gly Glu Glu Asp Val145 150 155 160Ile Tyr Thr Trp Lys Ala Leu
Gly Gln Ala Ala Asn Glu Ser His Asn 165 170 175Gly Ser Ile Leu Pro
Ile Ser Trp Arg Trp Gly Glu Ser Asp Met Thr 180 185 190Phe Ile Cys
Val Ala Arg Asn Pro Val Ser Arg Asn Phe Ser Ser Pro 195 200 205Ile
Leu Ala Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser 210 215
220Ser Met Val2254224PRTArtificialmu75/hu25 4Met Ala Arg Phe Ser
Thr Tyr Ile Ile Phe Thr Ser Val Leu Cys Gln1 5 10 15Leu Thr Val Thr
Ala Ala Ser Gly Thr Leu Lys Lys Val Ala Gly Ala 20 25 30Leu Asp Gly
Ser Val Thr Phe Thr Leu Asn Ile Thr Glu Ile Lys Val 35 40 45Asp Tyr
Val Val Trp Thr Phe Asn Thr Phe Phe Leu Ala Met Val Lys 50 55 60Lys
Asp Gly Val Thr Ser Gln Ser Ser Asn Lys Glu Arg Ile Val Phe65 70 75
80Pro Asp Gly Leu Tyr Ser Met Lys Leu Ser Gln Leu Lys Lys Asn Asp
85 90 95Ser Gly Ala Tyr Arg Ala Glu Ile Tyr Ser Thr Ser Ser Gln Ala
Ser 100 105 110Leu Ile Gln Glu Tyr Val Leu His Val Tyr Lys His Leu
Ser Arg Pro 115 120 125Lys Val Thr Ile Asp Arg Gln Ser Asn Lys Asn
Gly Thr Cys Val Ile 130 135 140Asn Leu Thr Cys Ser Thr Asp Gln Asp
Gly Glu Asn Val Thr Tyr Ser145 150 155 160Trp Lys Ala Val Gly Gln
Ala Ala Asn Glu Ser His Asn Gly Ser Ile 165 170 175Leu Pro Ile Ser
Trp Arg Trp Gly Glu Ser Asp Met Thr Phe Ile Cys 180 185 190Val Ala
Arg Asn Pro Val Ser Arg Asn Phe Ser Ser Pro Ile Leu Ala 195 200
205Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser Ser Met Val
210 215 2205119PRTHomo sapiens 5Glu 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 Asp Phe Ser Arg Tyr 20 25 30Trp Met Ser Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn Pro Asp
Ser Ser Thr Ile Asn Tyr Ala Pro Ser Leu 50 55 60Lys Asp Lys Phe Ile
Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Pro Asp Gly Asn Tyr Trp Tyr Phe Asp Val Trp Gly Gln Gly 100 105
110Thr Leu Val Thr Val Ser Ser 1156107PRTHomo sapiens 6Asp Ile Gln
Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg
Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Ile Ala 20 25 30Val
Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile 35 40
45Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp 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 Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser
Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100
105
* * * * *