U.S. patent application number 14/139096 was filed with the patent office on 2014-10-30 for methods of treating multiple myeloma using combination therapies based on anti-cs1 antibodies.
The applicant listed for this patent is AbbVie Biotherapeutics Inc.. Invention is credited to Daniel Afar, Kenneth C. Anderson, Yu-Tzu Tai.
Application Number | 20140322201 14/139096 |
Document ID | / |
Family ID | 38831279 |
Filed Date | 2014-10-30 |
United States Patent
Application |
20140322201 |
Kind Code |
A1 |
Afar; Daniel ; et
al. |
October 30, 2014 |
METHODS OF TREATING MULTIPLE MYELOMA USING COMBINATION THERAPIES
BASED ON ANTI-CS1 ANTIBODIES
Abstract
Compositions and methods for treating MM are provided
herein.
Inventors: |
Afar; Daniel; (Short Hills,
NJ) ; Anderson; Kenneth C.; (Wellesley, MA) ;
Tai; Yu-Tzu; (Newton, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AbbVie Biotherapeutics Inc. |
Redwood City |
CA |
US |
|
|
Family ID: |
38831279 |
Appl. No.: |
14/139096 |
Filed: |
December 23, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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11835257 |
Aug 7, 2007 |
8632772 |
|
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14139096 |
|
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60836185 |
Aug 7, 2006 |
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60944262 |
Jun 15, 2007 |
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Current U.S.
Class: |
424/133.1 ;
424/139.1 |
Current CPC
Class: |
A61K 39/39558 20130101;
A61P 35/02 20180101; A61K 2039/542 20130101; C07K 16/3061 20130101;
C07K 2317/56 20130101; A61K 2039/505 20130101; A61K 9/0019
20130101; A61K 39/3955 20130101; A61K 31/454 20130101; A61P 35/00
20180101; A61K 38/05 20130101; A61K 45/06 20130101; A61K 31/454
20130101; C07K 2317/732 20130101; A61K 39/39558 20130101; A61K
2300/00 20130101; A61K 31/573 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 31/69 20130101; A61K 2300/00 20130101; C07K
2317/24 20130101; A61K 31/573 20130101; C07K 16/2806 20130101; A61K
31/69 20130101 |
Class at
Publication: |
424/133.1 ;
424/139.1 |
International
Class: |
C07K 16/28 20060101
C07K016/28; A61K 31/69 20060101 A61K031/69; A61K 31/573 20060101
A61K031/573; A61K 39/395 20060101 A61K039/395; A61K 31/454 20060101
A61K031/454 |
Claims
1-63. (canceled)
64. A method of treating multiple myeloma in a subject, comprising
administering to a subject in need thereof a first pharmaceutical
composition comprising a therapeutically effective amount of an
anti-CS1 antibody and a second pharmaceutical composition
comprising a therapeutically effective amount of lenalidomide.
65. The method of claim 64, wherein the anti-CS1 antibody binds the
epitope covered by amino acids 170 to 227 of SEQ ID NO:1.
66. The method of claim 65, wherein the anti-CS1 antibody further
binds the epitope covered by amino acids 23 to 151 of SEQ ID NO:1
or the epitope covered by amino acids 68 to 151 of SEQ ID NO:1.
67. The method of claim 64, wherein the anti-CS1 antibody is
HuLuc63, said HuLuc63 being a humanized IgG.sub.1 antibody
comprising the heavy chain variable region of SEQ ID NO:5 and the
light chain variable region of SEQ ID NO:6.
68. The method of claim 64, in which the anti-CS1 antibody is
administered intravenously at a dosage from approximately 0.5 mg/kg
to approximately 20 mg/kg.
69. The method of claim 67, in which the HuLuc63 is administered
intravenously at a dosage from approximately 0.5 mg/kg to
approximately 20 mg/kg.
70. The method of claim 68, in which lenalidomide is administered
orally at a dosage from approximately 1 mg/day to 50 mg/day.
71. A method of treating multiple myeloma in a subject, comprising
administering to a subject in need thereof a first pharmaceutical
composition comprising a therapeutically effective amount of an
anti-CS1 antibody, a second pharmaceutical composition comprising a
therapeutically effective amount of lenalidomide, and a
therapeutically effective amount of dexamethasone.
72. The method of claim 71, wherein the anti-CS1 antibody binds the
epitope covered by amino acids 170 to 227 of SEQ ID NO:1.
73. The method of claim 72, wherein the anti-CS1 antibody further
binds the epitope covered by amino acids 23 to 151 of SEQ ID NO:1
or the epitope covered by amino acids 68 to 151 of SEQ ID NO:1.
74. The method of claim 71, wherein the anti-CS1 antibody is
HuLuc63, said HuLuc63 comprising a humanized IgG.sub.1 antibody
comprising the heavy chain variable region of SEQ ID NO:5 and the
light chain variable region of SEQ ID NO:6.
75. The method of claim 71, in which the anti-CS1 antibody is
administered intravenously at a dosage from approximately 0.5 mg/kg
to approximately 20 mg/kg.
76. The method of claim 74, in which the HuLuc63 is administered
intravenously at a dosage from approximately 0.5 mg/kg to
approximately 20 mg/kg.
77. The method of claim 75, in which lenalidomide is administered
orally at a dosage from approximately 1 mg/day to 50 mg/day.
78. A method of treating multiple myeloma in a subject, comprising
administering to a subject in need thereof a first pharmaceutical
composition comprising a therapeutically effective amount of an
anti-CS1 antibody, a second pharmaceutical composition comprising a
therapeutically effective amount of lenalidomide, and a third
pharmaceutical composition comprising a therapeutically effective
amount of bortezomib.
79. The method of claim 78, wherein the anti-CS1 antibody binds the
epitope covered by amino acids 170 to 227 of SEQ ID NO:1.
80. The method of claim 79, wherein the anti-CS1 antibody further
binds the epitope covered by amino acids 23 to 151 of SEQ ID NO:1
or the epitope covered by amino acids 68 to 151 of SEQ ID NO:1.
81. The method of claim 78, wherein the anti-CS1 antibody is
HuLuc63, said HuLuc63 being a humanized IgG.sub.1 antibody
comprising the heavy chain variable region of SEQ ID NO:5 and the
light chain variable region of SEQ ID NO:6.
82. The method of claim 78, in which the anti-CS1 antibody is
administered intravenously at a dosage from approximately 0.5 mg/kg
to approximately 20 mg/kg.
83. The method of claim 81, in which the HuLuc63 is administered
intravenously at a dosage from approximately 0.5 mg/kg to
approximately 20 mg/kg.
84. The method of claim 82, in which lenalidomide is administered
orally at a dosage from approximately 1 mg/day to 50 mg/day.
85. A method of treating multiple myeloma in a subject, comprising
administering to a subject in need thereof a first pharmaceutical
composition comprising a therapeutically effective amount of an
anti-CS1 antibody, a second pharmaceutical composition comprising a
therapeutically effective amount of lenalidomide, a third
pharmaceutical composition comprising a therapeutically effective
amount of bortezomib, and a therapeutically effective amount of
dexamethasone.
86. The method of claim 85, wherein the anti-CS1 antibody binds the
epitope covered by amino acids 170 to 227 of SEQ ID NO:1.
87. The method of claim 86, wherein the anti-CS1 antibody further
binds the epitope covered by amino acids 23 to 151 of SEQ ID NO:1
or the epitope covered by amino acids 68 to 151 of SEQ ID NO:1.
88. The method of claim 85, wherein the anti-CS1 antibody is
HuLuc63, said HuLuc63 being a humanized IgG.sub.1 antibody
comprising the heavy chain variable region of SEQ ID NO:5 and the
light chain variable region of SEQ ID NO:6.
89. A method of treating multiple myeloma in a subject, comprising
administering to a subject in need thereof a first pharmaceutical
composition comprising a therapeutically effective amount of an
anti-CS1 antibody, a second pharmaceutical composition comprising a
therapeutically effective amount of bortezomib, and a
therapeutically effective amount of dexamethasone.
90. The method of claim 89, wherein the anti-CS1 antibody binds the
epitope covered by amino acids 170 to 227 of SEQ ID NO:1.
91. The method of claim 90, wherein the anti-CS1 antibody further
binds the epitope covered by amino acids 23 to 151 of SEQ ID NO:1
or the epitope covered by amino acids 68 to 151 of SEQ ID NO:1.
92. The method of claim 89, wherein the anti-CS1 antibody is
HuLuc63, said HuLuc63 being a humanized IgG.sub.1 antibody
comprising the heavy chain variable region of SEQ ID NO:5 and the
light chain variable region of SEQ ID NO:6.
Description
1. CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of application Ser. No.
11/835,257, filed Aug. 7, 2007, now U.S. Pat. No. 8,632,772, which
claims benefit under 35 U.S.C. .sctn.119(e) to application Ser. No.
60/836,185, filed Aug. 7, 2006 and 60/944,262, filed Jun. 15, 2007,
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 with MM 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
treatment have been tested (See Case, D C et al., (1977) Am. J. Med
63:897 903), but 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 and therapeutic agents, 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-subset1), 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, such as HuLuc63, in combination with one or more
therapeutic agents. The anti-CS1 antibody is typically administered
in a first pharmaceutical composition as an intravenous infusion at
doses ranging from 0.5 to 20 mg/kg, from once a week to once a
month.
[0006] A second pharmaceutical composition comprising one or more
therapeutic agents, such as bortezomib and lenalidomide, can be
administered concurrently, prior to, or following administration of
an anti-CS1 antibody. Depending on the agent, the composition can
be administered orally, intravenously or subcutaneously.
Therapeutic agents can be used at high dose rates, standard dose
rates and at reduced dose rates.
[0007] In some embodiments, administration of the pharmaceutical
compositions described herein increases the sensitivity of multiple
myeloma cells to a therapeutic agent. By way of example, but not
limitation, inclusion of an anti-CS1 antibody can enhance the
activity of the therapeutic agent, 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] FIGS. 1A-1C depict autologous ADCC-mediated lysis of MM
cells treated with HuLuc63;
[0010] FIGS. 2A-2B depict HuLuc63 induced ADCC against Hsp90 and
bortezomib resistant patient tumor cells;
[0011] FIGS. 3A-3C depict enhancement of HuLuc63 induced ADCC
against MM cells when effector cells were pretreated with
lenalidomide;
[0012] FIGS. 4A-4D depict the effect of bortezomib pre-treatment on
HuLuc63-mediated ADCC in vitro. Examples are shown for 4 different
donors; and,
[0013] FIGS. 5A-5B depict the effect of HuLuc63 and bortezomib in
OPM2 tumor-bearing mice.
5. DETAILED DESCRIPTION
[0014] 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).
[0015] 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:
[0016] (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. [0017] (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. [0018] (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.
[0019] 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 NO: 1 Tyr Ile Leu Trp Gln
Leu 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 NO: 2 Tyr Ile Leu Trp Gln Leu
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 NO: 3 Ser Val Leu Cys Gln Leu 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 NO: 4 Ser Val Leu
Cys Gln Leu 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 NO: 5 Val Gln Pro Gly Gly Ser 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 NO: 6 Ser Ala Ser Val Gly Asp 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 Trp 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
[0020] 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.
[0021] 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.
[0022] 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).
[0023] 5.2 Pharmaceutical Compositions
[0024] Provided herein are pharmaceutical compositions that are
beneficial in reducing tumor mass and/or regressing tumor growth,
in patients diagnosed with multiple myeloma. The components of the
pharmaceutical compositions are addressed in more detail below, but
typically include an anti-CS1 antibody, such as HuLuc63 and one or
more therapeutic agents. 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 combined with 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 HuLuc63 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 (MTD) (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,
an 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, an 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] 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 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).
[0033] 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%.
[0034] 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.
[0035] Examples of therapeutic agents that can be used in the
compositions described herein include, but are not limited to,
dexamethasone, thalidomide, melphalan, prednisone, doxorubicin,
doxorubicin HCL liposome injection, bortezomib, lenalidomide,
and/or combinations thereof.
[0036] 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
lenalidomide.
[0037] 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 bortezomib.
[0038] 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
lenalidomide and a therapeutically effective amount of bortezomib.
In some embodiments, lenalidomide and bortezomib 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 lenalidomide, and a third comprising
bortezomib.
[0039] 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
lenalidomide and dexamethasone. In some embodiments, lenalidomide
and dexamethasone 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
lenalidomide, and a third comprising dexamethasone.
[0040] 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 bortezomib and dexamethasone.
In some embodiments, bortezomib and dexamethasone 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 bortezomib, and a third comprising
dexamethasone.
[0041] 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 therapeutically effective amount of lenalidomide,
bortezomib, and dexamethasone. In some embodiments, lenalidomide,
bortezomib, and dexamethasone are provided separately. 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. 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 lenalidomide and
bortezomib.
[0042] 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 bortezomib
and optionally can comprise one or more of the following agents:
thalidomide, dexamethasone, melphalan, doxorubicin, doxorubicin HCl
liposome injection, and/or prednisone. Provided that the agents
retain their efficacy, compositions comprising various combinations
of thalidomide, dexamethasone, melphalan, doxorubicin, doxorubicin
HCl liposome injection, and prednisone 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.
[0043] 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, anti-CS1 antibodies can be packaged in dosages ranging
from about 1 to 1000 mg. In some embodiments, anti-CS1 antibodies
can be 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.
[0044] 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.
[0045] 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.
[0046] 5.3 Methods
[0047] The pharmaceutical compositions described herein find use in
treating MM. 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.
[0048] 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).
[0049] 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.
[0050] 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.
[0051] 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)).
[0052] 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.
[0053] In other embodiments, administration of the pharmaceutical
composition elicits a very good partial response or a partial
response.
[0054] In other embodiments, administration of the pharmaceutical
composition elicits a minimal response.
[0055] 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.
[0056] Routes of administration and dosage ranges for compositions
comprising an anti-CS1 antibody 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).
[0057] 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.
[0058] 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., minimal, partial, or complete, as
noted by the clinician or other qualified observer, and as defined
by the EBMT.
[0059] Generally, an anti-CS1 antibody, such as HuLuc63, 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).
[0060] In those embodiments in which 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.
[0061] 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).
[0062] 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 Lenalidomide
[0063] Lenalidomide is the first of a new class of oral cancer
drugs called IMiDs.RTM.. These immunomodulatory derivatives are
chemically similar to thalidomide but are more potent and have a
different side effect profile than thalidomide. They have multiple
mechanisms of action that affect both the cancer cell and its
microenvironment. Lenalidomide has been shown to induce immune
responses, enhance activity of immune cells, and inhibit
inflammation. For example, lenalidomide may enhance the activation
of T cells and NK cells, induce production of interleukin 2 and
inhibit pro-inflammatory cytokines such as tumor necrosis
factor-alpha and interleukin 1-beta. Currently lenalidomide in
combination with dexamethasone is approved for 2.sup.nd line
therapy of multiple myeloma.
[0064] In Vitro ADCC Assay: Methods and Results
[0065] ADCC was measured by calcein-AM release assay, with
sensitivity similar to traditional Cr.sup.51 assay, as described
previously. After informed consent, peripheral blood mononuclear
cells (PBMCs) including natural killer (NK) effector cells were
isolated from leukopheresis products of normal donors or peripheral
blood from MM patients. Increasing concentrations (0-10 .mu.g/ml)
of either HuLuc63 or human isotype control IgG.sub.1 MSL109 mAbs
were added at effector: target (E:T) ratios of 20:1, in a final
volume of 200 .mu.l per well. In some experiments, PBMC effector
cells were pretreated with lenalidomide for 3 days at 0.2 .mu.M
before HuLuc63-mediated ADCC assays were performed. After 4 h
incubation, 100 .mu.l culture supernatants were transferred to a
Black ViewPlate.TM.-96 plate and arbitrary fluorescent units (AFU)
were read on a fluorometer (Wallac VICTOR2). This assay is valid
only if (AFU mean maximum release-medium control release)/(AFU mean
spontaneous release-medium control release)>7. Calculation of %
specific lysis from triplicate experiments was done using the
following equation:
% Specific Lysis = 100 .times. ( AFU mean experimental release -
AFU mean spontaneous release 1 ) ( AFU mean maximal release 2 - AFU
mean spontaneous release ) ##EQU00001##
.sup.1 Calcein-AM release by target cells in the absence of Ab or
NK cells..sup.2 Calcein-AM release by target cells upon lysis by
detergent.
[0066] HuLuc63-mediated lysis of patient MM cells by effector cells
from the same patient was measured using an ADCC assay. HuLuc63,
but not iso IgG.sub.1, induced significant autologous myeloma cell
lysis in patients in patient samples (FIGS. 1A-1C).
HuLuc63-mediated autologous tumor cell lysis was also demonstrated
in patients with MM resistant or refractory to novel anti-MM
therapies including bortezomib and/or 17-AAG (targeting heat shock
protein 90) (FIGS. 2A and 2B). These data suggest that HuLuc63 can
target myeloma cells from patients that are newly diagnosed, or
resistant to standard of care drugs and/or novel agents.
[0067] HuLuc63-mediated lysis of patient myeloma cells by PBMC
effector cells from the same patient was measured using an ADCC
assay. CD138-purified tumor cells from a patient with MM were
incubated with autologous effector cells in the presence of serial
dilutions of HuLuc63 (solid symbols) or isotype control IgG.sub.1
(open symbols). PBMC effector cells were pre-incubated for 3 days
in the presence or absence of with lenalidomide (0.2 mM) (square
symbols) or vehicle control (round symbols), followed by
HuLuc63-mediated ADCC. HuLuc63-mediated ADCC, but not iso
IgG.sub.1, induced significant autologous myeloma cell lysis in a
patient sample. Pre-incubation of the PBMC effector cells with
lenalidomide significantly increased the ADCC activity (FIG. 3C).
Similarly, pretreatment of effector cells with lenalidomide
enhanced HuLuc63-induced lysis of myeloma cell lines (FIGS. 3A and
3B). These results provide the framework for a treatment strategy
combining lenalidomide with HuLuc63 in MM.
Example 2
HuLuc63 in Combination with Bortezomib
[0068] Bortezomib is a potent, specific, and reversible proteasome
inhibitor. Proteasomes are present in all cells and function to
help regulate cell growth. Inhibition of the proteasome results in
apoptosis of cancer cells. Bortezomib has been shown to be
particularly effective at killing myeloma cells and is currently
approved for 2.sup.nd and 3.sup.rd line therapy in multiple
myeloma. Recent data has shown that bortezomib treatment of myeloma
cells results in down-modulation of cell-surface expression of MHC
class I, an inhibitor of NK function (Shi et al., Blood (ASH Annual
Meeting Abstracts), November 2006; 108:3498). The hypothesis is
that bortezomib treatment of myeloma cells would make them more
susceptible to NK-mediated killing and, thus, enhance
HuLuc63-mediated ADCC. The purpose of this study was to examine
whether using HuLuc63 in combination with bortezomib provided
therapeutic benefit.
[0069] The effect of HuLuc63 and bortezomib treatment on expression
of CS1 in MM cell lines and mouse xenograft tumors was examined by
flow cytometry and immunohistochemistry respectively.
[0070] In Vitro ADCC Assay: Methods and Results
[0071] OPM2 myeloma cells were harvested at mid-log phase,
suspended at a density of 1.0.times.10.sup.6 cells/mL in complete
media (RPMI with 10% FBS) and treated overnight with or without
Velcade (10 nM). Cells were collected, washed, re-suspended at a
density of 20.times.106 viable cells/mL, and labeled for one hour
with 50 mCi Na.sub.2[.sup.51Cr]O.sub.4 per 10.sup.6 cells.
.sup.51Cr-Labeled cells were washed then added to a 96-well
V-bottomed polystyrene plate at a cell density of 15,000 cells per
75 .mu.L RPMI supplemented with 10% heat-inactivated FBS. HuLuc63
and a human IgG.sub.1 isotype control antibody MSL-109 were added
to target cells for a final antibody concentration ranging from
0.001 to 10 .mu.g/mL. NK cells were enriched from the whole blood
of healthy donors using the RosetteSep human NK cell enrichment
cocktail (Stem Cell Technologies). The enriched NK cells were added
to Velcade treated or untreated OPM2 cells at a ratio of 10:1.
After a 4-hour incubation at 37.degree. C., cells were centrifuged
and the supernatants measured for released .sup.51Cr. Maximum
release was determined from target cells lysed with 100 mg/ml
Digitonin. Antibody independent cellular cytotoxicity (AICC) was
determined using target cells, plus media, plus NK cells, while
spontaneous lysis was determined using .sup.51Cr-labelled cells
plus media without NK effectors.
% Cytotoxicity was calculated
as((sample-AICC)/(Maximum-AICC)*100.
[0072] CS1 protein expression was examined on the OPM2 multiple
myeloma cell line with no significant change in CS1 expression
observed pre- or post-treatment with HuLuc63, bortezomib or with
both agents. The combination of HuLuc63 with bortezomib was then
tested for anti-myeloma activity in vitro using ADCC assays. The
results showed that pre-treatment with bortezomib significantly
enhanced HuLuc63-mediated ADCC towards OPM2 cells using NK effector
cells from healthy donors. OPM2 cells were pretreated with vehicle
control (square symbols) or bortezomib (10 nM; round symbols) for
18 hrs and were then subjected to HuLuc63 mediated ADCC using human
NK cells from healthy donors. HuLuc63 (closed symbols) and isotype
control antibody (open symbols) were used at doses ranging from
0.001-10 .mu.g/ml. The results show that bortezomib pre-treatment
significantly decreased the EC.sub.50 for HuLuc63-mediated ADCC in
vitro (FIGS. 4A-4D, Table 3).
TABLE-US-00003 TABLE 3 No Bortezomib P value Treatment (10 nM) (t
test) 1 0.0758 0.0106 0.04 2 0.149 0.057 0.05 3 0.103 0.0459 0.004
4 0.0302 0.0207 0.0004
[0073] In Vivo Xenograft Mouse Model: Methods and Results
[0074] Six- to eight-week old female IcrTac:ICR-Prkdc.sup.scid mice
obtained from Taconic Farms (Germantown, N.Y.) were inoculated with
1.times.10.sup.7 OPM2 (American Type Culture Collection) cells 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 mg/kg of antibody administered
intraperitoneally twice a week for a total of 6 doses. Bortezomib
was administered intraperitoneally at a dose of 0.75 mg/kg twice a
week for a total of 6 doses. 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.
[0075] To examine the effect of HuLuc63 combination therapy with
bortezomib 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. Bortezomib was given twice a week at
0.75 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 bortezomib (FIG. 5A). Mice in
the combination treatment group exhibited on average 40-50% smaller
tumors than in the HuLuc63 monotherapy group, and 60-70% smaller
tumors than in the bortezomib group.
[0076] In a second experiment, HuLuc63 was combined with bortezomib
in vivo, using a different dose and dosing schedule for bortezomib,
while keeping the original HuLuc63 dose and dosing schedule. OPM2
cells were inoculated into the flanks of SCID mice. When tumors
reached an average size of about 100 mm3, animals were randomized
into 4 groups of 15 mice each and were treated with 1 mg/kg of
antibody administered intraperitoneally twice a week for a total of
10 doses. Bortezomib was administered intraperitoneally at a dose
of 1 mg/kg twice for weeks 1 and 2, no treatment for week 3, and 1
mg/kg twice for weeks 4 and 5 for a total of 8 doses. The intent
for this dosing schedule was to more closely mimic the dosing
schedule of bortezomib in the clinic, where each treatment cycle
consists of 2 weeks of dosing, with one week off. Tumor growth was
monitored for a period of 1-2 months.
[0077] The results showed significant anti-tumor activity of
HuLuc63 alone, bortezomib alone and for HuLuc63 in combination with
bortezomib (FIG. 5B). Mice in the combination treatment group
exhibited significantly smaller tumors than mice treated with
either drug alone. The data indicates that bortezomib synergizes
with HuLuc63 in anti-myeloma tumor activity.
Example 3
Phase 1b, Open-Label, Dose-Escalation Study of HuLuc63 and
Bortezomib in Multiple Myeloma Patients Following First or Second
Relapse
[0078] The proposed Phase 1b, multi-center, open-label, multi-dose,
dose escalation study will evaluate the combination of HuLuc63 and
bortezomib in patients with multiple myeloma after 1st or 2nd
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 a fixed dose of bortezomib IV at 1.0 mg/m2.
Patients will receive HuLuc63 every 10 days and bortezomib will be
given in 21-day cycles (twice weekly for two weeks (days 1, 4, 8,
11) followed by a 10-day rest period (days 12-21)).
[0079] After 9 weeks of therapy (6 doses of HuLuc63, 3 cycles of
bortezomib), EBMT criteria will be assessed. If a patient has
progressive disease, HuLuc63 will be discontinued and bortezomib
may be withdrawn or continued at the discretion of the site
investigator. If the patient has responded or has stable disease at
Week 9, dosing with HuLuc63 and bortezomib will continue so that a
total of 24 weeks of treatment (16 doses HuLuc63, 8 cycles
bortezomib) are completed or disease progression occurs. Dosing
with HuLuc63 and bortezomib will continue until the data from the
Week 9 visit are available.
[0080] Patients will receive HuLuc63 IV once every 10 days, with
each dose infused over 1 hour. Bortezomib will be given as IVP for
8 three-week cycles with each cycle consisting of bortezomib on
days 1, 4, 8 and 11 followed by a ten-day rest period (days 11-21).
Dosing cohorts are as follows: 2.5 mg/kg HuLuc63/1.3 mg/m.sup.2
bortezomib; 5 mg/kg HuLuc63/1.3 mg/m.sup.2 bortezomib; 10 mg/kg
HuLuc63/1.3 mg/m.sup.2 bortezomib; 15 mg/kg HuLuc63/1.3 mg/m.sup.2
bortezomib; and, 20 mg/kg HuLuc63/1.3 mg/m.sup.2 bortezomib.
[0081] HuLuc63 will be provided at a concentration of 10 mg/mL in
an intravenous formulation in vials. Bortezomib will be provided as
a 3.5 mg lyophilized cake or powder in a 10 mL vial, to be
reconstituted with 3.5 mL normal (0.9%) saline, sodium chloride
injection to 3.5 mL of 1 mg/mL of bortezomib, as per Velcade.RTM.
package insert.
[0082] Approximately 15 to 30 patients in 5 cohorts will be
enrolled in the trial. Each cohort will begin with 3 patients. If
no dose-limiting toxicity (DLT) is noted within the first 6 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.
[0083] 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
bortezomib. For dose escalation to the next cohort, 3 assessable
patients must complete their first 6 weeks (4 doses HuLuc63, 2
cycles bortezomib). 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 .ltoreq.33%. The highest tolerated
dose will be HuLuc63 20 mg/kg+bortezomib 1.0 mg/m.sup.2 if no dose
limiting toxicities are observed.
Example 4
Phase 1b, Multi-Center, Open-Label, Dose-Escalation Study of
HuLuc63 and Lenalidomide
[0084] The proposed Phase 1b, multi-center, open-label, multi-dose,
dose escalation study will evaluate the combination of HuLuc63 and
lenalidomide in patients with multiple myeloma after 1st or 2nd
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 a fixed dose of lenalidomide PO at 15 mg. Patients
will receive HuLuc63 every 7 days and lenalidomide will be given in
28-day cycles (once daily for 21 days followed by a 7-day rest
period (days 22-28)).
[0085] After 8 weeks of therapy (8 doses of HuLuc63, 2 cycles of
lenalidomide), EBMT criteria will be assessed. Dexamethasone will
be added to the regimen at an oral dose of 40 mg daily on days 1,
8, 15 and 22 of a 4-week cycle. If at week 12 (12 doses HuLuc63, 3
cycles of lenalidomide, 1 cycle of dexamethasone) there is evidence
of progressive disease, HuLuc63 will be stopped and lenalidomide
and dexamethasone will be continued up to 16 weeks at the
discretion of the investigator. If a patient has stable disease or
better, they will continue on HuLuc63 until week 16 (15 total
doses) or disease progression. EMBT criteria will be evaluated at
week 16.
[0086] Patients will receive HuLuc63 IV once every 10 days, with
each dose infused over 1 hour. Lenalidomide will be given orally
daily for 3 weeks followed by a weeklong rest period. Dosing
cohorts are as follows: 2.5 mg/kg HuLuc63/15 mg lenalidomide; 5
mg/kg HuLuc63/15 mg lenalidomide; 10 mg/kg HuLuc63/15 mg
lenalidomide; 15 mg/kg HuLuc63/15 mg lenalidomide; and, 20 mg/kg
HuLuc63/15 mg lenalidomide. After week 8, dexamethasone will be
added to the above regimens at an oral dose of 40 mg daily on days
1, 8, 15 and 22 of a 4 week cycle.
[0087] HuLuc63 will be provided at a concentration of 10 mg/mL in
an intravenous formulation in vials. Lenalidomide will be supplied
as 5 mg and 10 mg capsules for oral administration.
[0088] Approximately 15 to 30 patients in 5 cohorts will be
enrolled in the trial. 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.
[0089] 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
lenalidomide. For dose escalation to the next cohort, 3 assessable
patients must complete their first 4 weeks (4 doses HuLuc63, 1
cycle lenalidomide). 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 .ltoreq.33%. The highest
tolerated dose will be HuLuc63 20 mg/kg+lenalidomide 15 mg if no
dose limiting toxicities are observed.
[0090] 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.
[0091] 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 Gln 1 5 10 15 Leu Thr Gly Ser Ala Ala Ser Gly Pro
Val Lys Glu Leu Val Gly Ser 20 25 30 Val Gly Gly Ala Val Thr Phe
Pro Leu Lys Ser Lys Val Lys Gln Val 35 40 45 Asp Ser Ile Val Trp
Thr Phe Asn Thr Thr Pro Leu Val Thr Ile Gln 50 55 60 Pro Glu Gly
Gly Thr Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg 65 70 75 80 Val
Asp Phe Pro Asp Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys 85 90
95 Lys Asn Asp Ser Gly Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu
100 105 110 Gln Gln Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu
His Leu 115 120 125 Ser Lys Pro Lys Val Thr Met Gly Leu Gln Ser Asn
Lys Asn Gly Thr 130 135 140 Cys Val Thr Asn Leu Thr Cys Cys Met Glu
His Gly Glu Glu Asp Val 145 150 155 160 Ile Tyr Thr Trp Lys Ala Leu
Gly Gln Ala Ala Asn Glu Ser His Asn 165 170 175 Gly Ser Ile Leu Pro
Ile Ser Trp Arg Trp Gly Glu Ser Asp Met Thr 180 185 190 Phe Ile Cys
Val Ala Arg Asn Pro Val Ser Arg Asn Phe Ser Ser Pro 195 200 205 Ile
Leu Ala Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser 210 215
220 Ser Met Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu
225 230 235 240 Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu
Arg Gln Glu 245 250 255 Glu Tyr Ile Glu Glu Lys Lys Arg Val Asp Ile
Cys Arg Glu Thr Pro 260 265 270 Asn Ile Cys Pro His Ser Gly Glu Asn
Thr Glu Tyr Asp Thr Ile Pro 275 280 285 His Thr Asn Arg Thr Ile Leu
Lys Glu Asp Pro Ala Asn Thr Val Tyr 290 295 300 Ser Thr Val Glu Ile
Pro Lys Lys Met Glu Asn Pro His Ser Leu Leu 305 310 315 320 Thr Met
Pro Asp Thr Pro Arg Leu Phe Ala Tyr Glu Asn Val Ile 325 330 335
2227PRTArtificialhu50/mu50 amino acids 1-151 of human CS1 fused to
amino acids 149-224 of mouse CS1 2Met Ala Gly Ser Pro Thr Cys Leu
Thr Leu Ile Tyr Ile Leu Trp Gln 1 5 10 15 Leu Thr Gly Ser Ala Ala
Ser Gly Pro Val Lys Glu Leu Val Gly Ser 20 25 30 Val Gly Gly Ala
Val Thr Phe Pro Leu Lys Ser Lys Val Lys Gln Val 35 40 45 Asp Ser
Ile Val Trp Thr Phe Asn Thr Thr Pro Leu Val Thr Ile Gln 50 55 60
Pro Glu Gly Gly Thr Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg 65
70 75 80 Val Asp Phe Pro Asp Gly Gly Tyr Ser Leu Lys Leu Ser Lys
Leu Lys 85 90 95 Lys Asn Asp Ser Gly Ile Tyr Tyr Val Gly Ile Tyr
Ser Ser Ser Leu 100 105 110 Gln Gln Pro Ser Thr Gln Glu Tyr Val Leu
His Val Tyr Glu His Leu 115 120 125 Ser Lys Pro Lys Val Thr Ile Asp
Arg Gln Ser Asn Lys Asn Gly Thr 130 135 140 Cys Val Ile Asn Leu Thr
Cys Ser Thr Asp Gln Asp Gly Glu Asn Val 145 150 155 160 Thr Tyr Ser
Trp Lys Ala Val Gly Gln Gly Asp Asn Gln Phe His Asp 165 170 175 Gly
Ala Thr Leu Ser Ile Ala Trp Arg Ser Gly Glu Lys Asp Gln Ala 180 185
190 Leu Thr Cys Met Ala Arg Asn Pro Val Ser Asn Ser Phe Ser Thr Pro
195 200 205 Val Phe Pro Gln Lys Leu Cys Glu Asp Ala Ala Thr Asp Leu
Thr Ser 210 215 220 Leu Arg Gly 225 3227PRTArtificialmu25/hu75
amino acids 1-67 of mouse CS1 fused to amino acids 68-227 of human
CS1 3Met Ala Arg Phe Ser Thr Tyr Ile Ile Phe Thr Ser Val Leu Cys
Gln 1 5 10 15 Leu Thr Val Thr Ala Ala Ser Gly Thr Leu Lys Lys Val
Ala Gly Ala 20 25 30 Leu Asp Gly Ser Val Thr Phe Thr Leu Asn Ile
Thr Glu Ile Lys Val 35 40 45 Asp Tyr Val Val Trp Thr Phe Asn Thr
Phe Phe Leu Ala Met Val Lys 50 55 60 Lys Asp Gly Gly Thr Ile Ile
Val Thr Gln Asn Arg Asn Arg Glu Arg 65 70 75 80 Val Asp Phe Pro Asp
Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys 85 90 95 Lys Asn Asp
Ser Gly Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu 100 105 110 Gln
Gln Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu His Leu 115 120
125 Ser Lys Pro Lys Val Thr Met Gly Leu Gln Ser Asn Lys Asn Gly Thr
130 135 140 Cys Val Thr Asn Leu Thr Cys Cys Met Glu His Gly Glu Glu
Asp Val 145 150 155 160 Ile Tyr Thr Trp Lys Ala Leu Gly Gln Ala Ala
Asn Glu Ser His Asn 165 170 175 Gly Ser Ile Leu Pro Ile Ser Trp Arg
Trp Gly Glu Ser Asp Met Thr 180 185 190 Phe Ile Cys Val Ala Arg Asn
Pro Val Ser Arg Asn Phe Ser Ser Pro 195 200 205 Ile Leu Ala Arg Lys
Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser 210 215 220 Ser Met Val
225 4224PRTArtificialmu75/hu25 amino acids 1-166 of mouse CS1 fused
to amino acids 170-227 of human CS1 4Met Ala Arg Phe Ser Thr Tyr
Ile Ile Phe Thr Ser Val Leu Cys Gln 1 5 10 15 Leu Thr Val Thr Ala
Ala Ser Gly Thr Leu Lys Lys Val Ala Gly Ala 20 25 30 Leu Asp Gly
Ser Val Thr Phe Thr Leu Asn Ile Thr Glu Ile Lys Val 35 40 45 Asp
Tyr Val Val Trp Thr Phe Asn Thr Phe Phe Leu Ala Met Val Lys 50 55
60 Lys Asp Gly Val Thr Ser Gln Ser Ser Asn Lys Glu Arg Ile Val Phe
65 70 75 80 Pro Asp Gly Leu Tyr Ser Met Lys Leu Ser Gln Leu Lys Lys
Asn Asp 85 90 95 Ser Gly Ala Tyr Arg Ala Glu Ile Tyr Ser Thr Ser
Ser Gln Ala Ser 100 105 110 Leu Ile Gln Glu Tyr Val Leu His Val Tyr
Lys His Leu Ser Arg Pro 115 120 125 Lys Val Thr Ile Asp Arg Gln Ser
Asn Lys Asn Gly Thr Cys Val Ile 130 135 140 Asn Leu Thr Cys Ser Thr
Asp Gln Asp Gly Glu Asn Val Thr Tyr Ser 145 150 155 160 Trp Lys Ala
Val Gly Gln Ala Ala Asn Glu Ser His Asn Gly Ser Ile 165 170 175 Leu
Pro Ile Ser Trp Arg Trp Gly Glu Ser Asp Met Thr Phe Ile Cys 180 185
190 Val Ala Arg Asn Pro Val Ser Arg Asn Phe Ser Ser Pro Ile Leu Ala
195 200 205 Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser Ser
Met Val 210 215 220 5119PRTHomo saiens 5Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Asp Phe Ser Arg Tyr 20 25 30 Trp Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly
Glu Ile Asn Pro Asp Ser Ser Thr Ile Asn Tyr Ala Pro Ser Leu 50 55
60 Lys Asp Lys Phe Ile Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Pro Asp Gly Asn Tyr Trp Tyr Phe Asp Val
Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 6
107PRTHomo sapiens 6Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser
Gln Asp Val Gly Ile Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro
Gly Lys Val Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg
His Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp
Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Tyr 85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
* * * * *