U.S. patent application number 16/376394 was filed with the patent office on 2020-02-27 for combination of anti-lag-3 antibodies and anti-pd-1 antibodies to treat tumors.
This patent application is currently assigned to Bristol-Myers Squibb Company. The applicant listed for this patent is Bristol-Myers Squibb Company. Invention is credited to David J. Fontana, Andres A. Gutierrez, Alan J. Korman, Katherine Lewis, Nils Lonberg, Mark J. Selby.
Application Number | 20200062845 16/376394 |
Document ID | / |
Family ID | 51743546 |
Filed Date | 2020-02-27 |
United States Patent
Application |
20200062845 |
Kind Code |
A1 |
Korman; Alan J. ; et
al. |
February 27, 2020 |
COMBINATION OF ANTI-LAG-3 ANTIBODIES AND ANTI-PD-1 ANTIBODIES TO
TREAT TUMORS
Abstract
Provided are methods for clinical treatment of tumors (e.g.,
advanced solid tumors) using an anti-LAG-3 antibody in combination
with an anti-PD-1 antibody.
Inventors: |
Korman; Alan J.; (Piedmont,
CA) ; Lonberg; Nils; (Woodside, CA) ; Fontana;
David J.; (Clyde Hill, WA) ; Gutierrez; Andres
A.; (Lawrenceville, NJ) ; Selby; Mark J.; (San
Francisco, CA) ; Lewis; Katherine; (Lake Forest Park,
WA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Bristol-Myers Squibb Company |
Princeton |
NJ |
US |
|
|
Assignee: |
Bristol-Myers Squibb
Company
Princeton
NJ
|
Family ID: |
51743546 |
Appl. No.: |
16/376394 |
Filed: |
April 5, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16108973 |
Aug 22, 2018 |
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16376394 |
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15021102 |
Mar 10, 2016 |
10081681 |
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PCT/US2014/056277 |
Sep 18, 2014 |
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16108973 |
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62014471 |
Jun 19, 2014 |
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61880606 |
Sep 20, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07K 2317/94 20130101;
C07K 2317/565 20130101; A61K 2039/545 20130101; A61K 2039/507
20130101; A61K 2039/505 20130101; C07K 16/2818 20130101; C07K
2317/24 20130101; C07K 2317/76 20130101; C07K 16/2803 20130101;
A61K 2039/54 20130101; C07K 2317/53 20130101; C07K 2317/21
20130101; A61P 35/00 20180101; C07K 2317/56 20130101 |
International
Class: |
C07K 16/28 20060101
C07K016/28; A61P 35/00 20060101 A61P035/00 |
Claims
1-20. (canceled)
21. A kit for treating a solid tumor in a human patient, the kit
comprising: (a) an 80 mg dose of an anti-LAG-3 antibody comprising
CDR1, CDR2 and CDR3 domains of the heavy chain variable region
having the sequence set forth in SEQ ID NO:3, and CDR1, CDR2 and
CDR3 domains of the light chain variable region having the sequence
set forth in SEQ ID NO:5; (b) a 240 mg dose of an anti-PD-1
antibody comprising CDR1, CDR2 and CDR3 domains of the heavy chain
variable region having the sequence set forth in SEQ ID NO: 19, and
CDR1, CDR2 and CDR3 domains of the light chain variable region
having the sequence set forth in SEQ ID NO:21; and (c) instructions
for using the anti-LAG-3 antibody and anti-PD-1 antibody in a
method for treating a solid tumor.
22. (canceled)
23. (canceled)
24. The kit of claim 21, wherein the anti-PD-1 and anti-LAG-3
antibodies are formulated for intravenous administration.
25. The kit of claim 21, wherein the anti-PD-1 and anti-LAG-3
antibodies are formulated together.
26. The kit of claim 21, wherein the anti-PD-1 and anti-LAG-3
antibodies are formulated separately.
27. The kit of claim 21, wherein the solid tumor is chosen from
melanoma, non-small cell lung cancer (NSCLC), human papilloma virus
(HPV)-related tumor, and gastric adenocarcinoma.
28. The kit of claim 21, wherein the anti-LAG-3 antibody comprises
(a) a heavy chain variable region CDR1 comprising the sequence set
forth in SEQ ID NO:7; (b) a heavy chain variable region CDR2
comprising the sequence set forth in SEQ ID NO:8; (c) a heavy chain
variable region CDR3 comprising the sequence set forth in SEQ ID
NO:9; (d) a light chain variable region CDR1 comprising the
sequence set forth in SEQ ID NO:10; (e) a light chain variable
region CDR2 comprising the sequence set forth in SEQ ID NO:11; and
(f) a light chain variable region CDR3 comprising the sequence set
forth in SEQ ID NO:12.
29. The kit of claim 21, wherein the anti-LAG-3 antibody comprises
heavy and light chain variable regions comprising the sequences set
forth in SEQ ID NOs:3 and 5, respectively.
30. The kit of claim 21, wherein the anti-LAG-3 antibody comprises
heavy and light chains comprising the sequences set forth in SEQ ID
NOs: 1 and 2, respectively.
31. The kit of claim 21, wherein the anti-PD-1 antibody comprises
(a) a heavy chain variable region CDR1 comprising the sequence set
forth in SEQ ID NO:23; (b) a heavy chain variable region CDR2
comprising the sequence set forth in SEQ ID NO:24; (c) a heavy
chain variable region CDR3 comprising the sequence set forth in SEQ
ID NO:25; (d) a light chain variable region CDR1 comprising the
sequence set forth in SEQ ID NO:26; (e) a light chain variable
region CDR2 comprising the sequence set forth in SEQ ID NO:27; and
(f) a light chain variable region CDR3 comprising the sequence set
forth in SEQ ID NO:28.
32. The kit of claim 21, wherein the anti-PD-1 antibody comprises
heavy and light chain variable regions comprising the sequences set
forth in SEQ ID NOs: 19 and 21, respectively.
33. The kit of claim 21, wherein the anti-PD-1 antibody comprises
heavy and light chains comprising the sequences as set forth in SEQ
ID NOs: 17 and 18, respectively.
34. The kit of claim 21, wherein the anti-LAG-3 antibody and/or
anti-PD-1 antibody is a full-length antibody.
35. The kit of claim 21, wherein the anti-LAG-3 antibody has a
S228P mutation
36. The kit of claim 21, wherein the patient has been previously
treated with anti-PD-1 or anti-PD-L1 antibody therapy.
37. The kit of claim 21, further comprising a dose of an
anti-CTLA-4 antibody.
38. The kit of claim 37, wherein the anti-CTLA-4 antibody is
ipilimumab.
Description
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional
Application No. 61/880,606, filed Sep. 20, 2013 and U.S.
Provisional Application No. 62/014,471, filed Jun. 19, 2014. The
contents of any patents, patent applications, and references cited
throughout this specification are hereby incorporated by reference
in their entireties.
BACKGROUND
[0002] Lymphocyte activation gene-3 (LAG-3; CD223) is a type I
transmembrane protein that is expressed on the cell surface of
activated CD4.sup.+ and CD8.sup.+ T cells and subsets of NK and
dendritic cells (Triebel F, et al., J. Exp. Med. 1990;
171:1393-1405; Workman C T, et al., J. Immunol. 2009;
182(4):1885-91). LAG-3 is closely related to CD4, which is a
co-receptor for T helper cell activation. Both molecules have 4
extracellular Ig-like domains and require binding to their ligand,
major histocompatibility complex (MHC) class II, for their
functional activity. In contrast to CD4, LAG-3 is only expressed on
the cell surface of activated T cells and its cleavage from the
cell surface terminates LAG-3 signaling. LAG-3 can also be found as
a soluble protein but it does not bind to MHC class II and its
function is unknown.
[0003] It has been reported that LAG-3 plays an important role in
promoting regulatory T cell (Treg) activity and in negatively
regulating T cell activation and proliferation (Workman C J, et
al., J. Immunol. 2005; 174:688-695). Both natural and induced Treg
express increased LAG-3, which is required for their maximal
suppressive function (Camisaschi C, et al., J. Immunol. 2010;
184:6545-6551 and Huang C T, et al., Immunity. 2004; 21:503-513).
Furthermore, ectopic expression of LAG-3 on CD4.sup.+ effector T
cells reduced their proliferative capacity and conferred on them
regulatory potential against third party T cells (Huang C T, et
al., Immunity. 2004; 21:503-513). Recent studies have also shown
that high LAG-3 expression on exhausted lymphocytic
choriomeningitis virus (LCMV)-specific CD8.sup.+ T cells
contributes to their unresponsive state and limits CD8.sup.+ T cell
antitumor responses (Blackburn S D, et al., Nat. Immunol. 2009;
10:29-37 and Grosso J F, et al., J. Clin. Invest. 2007;
117:3383-3392). In fact, LAG-3 maintained tolerance to self and
tumor antigens via direct effects on CD8.sup.+ T cells in 2 murine
models (Grosso J F, et al., J. Clin. Invest. 2007;
117:3383-3392).
[0004] Immune tolerance observed in the setting of tumor
development and tumor recurrence, however, seems to be mediated by
the co-expression of various T cell negative regulatory receptors,
not solely from LAG-3. Data from chronic viral infection models
(Blackburn S D, et al., Nat. Immunol. 2009; 10:29-37, Grosso J F,
et al., J. Clin. Invest. 2007; 117:3383-3392, and Lyford-Pike S, et
al., Cancer Res. 2013; 73(6):1733-41), knock-out mice (Woo S R, et
al., Cancer Res. 2012; 72:917-927; Okazaki T, et al., J. Exp Med.
2011; 208:395-407, and Bettini M. et al., J. Immunol. 2011;
187:3493-3498), tumor recurrence models (Goding S R, et al., J.
Immunol. 2013; 190(9):4899-4909) and, to a more limited extent,
human cancer patients (Goding S R, et al., J. Immunol. 2013;
190(9):4899-4909, Matsuzaki J, et al., Proc. Natl. Acad. Sci., USA.
2010; 107:7875-7880, and Gandhi M K, et al., Blood. 2006;
108:2280-2289) support a model wherein T cells that are
continuously exposed to antigen become progressively inactivated
through a process termed "exhaustion." Exhausted T cells are
characterized by the expression of T cell negative regulatory
receptors, predominantly CTLA-4, PD-1, and LAG-3, whose action is
to limit the cell's ability to proliferate, produce cytokines, and
kill target cells and/or to increase Treg activity. However, the
timing and sequence of expression of these molecules in the
development and recurrence of tumors have not been fully
characterized.
[0005] Programmed Cell Death 1 (PD-1) is a cell surface signaling
receptor that plays a critical role in the regulation of T cell
activation and tolerance (Keir M E, et al., Annu Rev Immunol 2008;
26:677-704). It is a type I transmembrane protein and together with
BTLA, CTLA-4, ICOS and CD28, comprise the CD28 family of T cell
co-stimulatory receptors. PD-1 is primarily expressed on activated
T cells, B cells, and myeloid cells (Dong H, et al., Nat Med. 1999;
5:1365-1369). It is also expressed on natural killer (NK) cells
(Terme M, et al., Cancer Res 2011; 71:5393-5399). Binding of PD-1
by its ligands, PD-L1 and PD-L2, results in phosphorylation of the
tyrosine residue in the proximal intracellular immune receptor
tyrosine inhibitory domain, followed by recruitment of the
phosphatase SHP-2, eventually resulting in down-regulation of T
cell activation. One important role of PD-1 is to limit the
activity of T cells in peripheral tissues at the time of an
inflammatory response to infection, thus limiting the development
of autoimmunity (Pardoll D M., Nat Rev Cancer 2012; 12:252-264).
Evidence of this negative regulatory role comes from the finding
that PD-1-deficient mice develop lupus-like autoimmune diseases
including arthritis and nephritis, along with cardiomyopathy
(Nishimura H, et al., Immunity, 1999; 11:141-151; and Nishimura H,
et al., Science, 2001; 291:319-322). In the tumor setting, the
consequence is the development of immune resistance within the
tumor microenvironment. PD-1 is highly expressed on
tumor-infiltrating lymphocytes, and its ligands are up-regulated on
the cell surface of many different tumors (Dong H, et al., Nat Med
2002; 8:793-800). Multiple murine cancer models have demonstrated
that binding of ligand to PD-1 results in immune evasion. In
addition, blockade of this interaction results in anti-tumor
activity (Topalian S L, et al. NEJM 2012; 366(26):2443-2454; Hamid
O, et al., NEJM 2013; 369:134-144). Moreover, it has been shown
that inhibition of the PD-1/PD-L1 interaction mediates potent
antitumor activity in preclinical models (U.S. Pat. Nos. 8,008,449
and 7,943,743).
[0006] Patients with metastatic or refractory solid tumors have
very poor prognosis (Rosenberg S A, et al., Cancer immunotherapy in
Cancer: Principles & Practice of Oncology (Eds DeVita V T,
Lawrence T S and Rosenberg S A) 2011; 332-344 (Lippincott Williams
& Wilkins, Philadelphia Pa.)). Despite advances in multimodal
therapy, increases in overall survival in this patient population
have been limited. Accordingly, it is an object of the present
invention to provide improved methods for treating subjects with
such tumors (e.g., advanced refractory solid tumors).
SUMMARY
[0007] Provided herein are methods for treating tumors in a human
patient, particularly solid tumors (e.g., advanced refractory solid
tumors), comprising administering to the patient a combination of
an anti-LAG-3 antibody and an anti-PD-1 antibody, wherein the
combination is administered (or is for administration) according to
a particular clinical dosage regimen (i.e., at a particular dose
amount and according to a specific dosing schedule). In one
embodiment, the human patient suffers from melanoma, non-small cell
lung cancer (NSCLC), virally-related cancer, head and neck cancer
(HNC) or gastric adenocarcinoma.
[0008] An exemplary anti-LAG-3 antibody is BMS-986016 comprising
heavy and light chains comprising the sequences shown in SEQ ID
NOs:1 and 2, respectively, or antigen binding fragments and
variants thereof. In other embodiments, the antibody comprises the
heavy and light chain complementarity determining regions (CDRs) or
variable regions (VRs) of BMS-986016. Accordingly, in one
embodiment, the antibody comprises CDR1, CDR2, and CDR3 domains of
the heavy chain variable (VH) region of BMS-986016 having the
sequence shown in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains of
the light chain variable (VL) region of BMS-986016 having the
sequence shown in SEQ ID NO:5. In another embodiment, the antibody
comprises CDR1, CDR2 and CDR3 heavy chain sequences set forth in
SEQ ID NOs:7, 8, and 9, respectively, and CDR1, CDR2 and CDR3 light
chain sequences as set forth in SEQ ID NOs:10, 11, and 12,
respectively. In another embodiment, the antibody has VH and/or VL
regions comprising the amino acid sequences set forth in SEQ ID
NO:3 and/or SEQ ID NO:5, respectively. In another embodiment, the
antibody comprises the VH and/or VL regions encoded by the nucleic
acid sequences set forth in SEQ ID NO:4 and/or SEQ ID NO:6,
respectively. In another embodiment, the antibody competes for
binding with, and/or binds to the same epitope on LAG-3 as, the
above-mentioned antibodies. In another embodiment, the antibody has
at least about 90% variable region amino acid sequence identity
with the above-mentioned antibodies (e.g., at least about 90%, 95%
or 99% variable region identity with SEQ ID NO:3 or SEQ ID
NO:5).
[0009] An exemplary anti-PD-1 antibody is Nivolumab (also referred
to as "5C4" in WO 2006/121168; and known as BMS-936558, MDX-1106
and ONO-4538) comprising heavy and light chains comprising the
sequences shown in SEQ ID NOs:17 and 18, respectively, or antigen
binding fragments and variants thereof. In other embodiments, the
antibody comprises the heavy and light chain CDRs or VRs of
BMS-936558. Accordingly, in one embodiment, the antibody comprises
CDR1, CDR2, and CDR3 domains of the VH region of BMS-936558 having
the sequence shown in SEQ ID NO:19, and CDR1, CDR2 and CDR3 domains
of the VL region of BMS-936558 having the sequence shown in SEQ ID
NO:21. In another embodiment, the antibody comprises heavy chain
CDR1, CDR2 and CDR3 domains comprising the sequences set forth in
SEQ ID NOs:23, 24, and 25, respectively, and light chain CDR1, CDR2
and CDR3 domains comprising the sequences set forth in SEQ ID NOs:
26, 27, and 28, respectively. In another embodiment, the antibody
comprises VH and/or VL regions comprising the amino acid sequences
set forth in SEQ ID NO: 19 and/or SEQ ID NO:21, respectively. In
another embodiment, the antibody comprises the heavy chain variable
(VH) and/or light chain variable (VL) regions encoded by the
nucleic acid sequences set forth in SEQ ID NO:20 and/or SEQ ID
NO:22, respectively. In another embodiment, the antibody competes
for binding with, and/or binds to the same epitope on PD-1 as, the
above-mentioned antibodies. In another embodiment, the antibody has
at least about 90% variable region amino acid sequence identity
with the above-mentioned antibodies (e.g., at least about 90%, 95%
or 99% variable region identity with SEQ ID NO:19 or SEQ ID
NO:21).
[0010] Accordingly, in one aspect, methods of treating solid tumors
(e.g., advanced refractory solid tumors) in a human patient are
provided, the methods comprising administering to the patient, an
effective amount of each of:
[0011] (a) an anti-LAG-3 antibody comprising CDR1, CDR2 and CDR3
domains of the heavy chain variable region having the sequence set
forth in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains of the light
chain variable region having the sequence set forth in SEQ ID
NO:5,
[0012] (b) an anti-PD-1 antibody comprising CDR1, CDR2 and CDR3
domains of the heavy chain variable region having the sequence set
forth in SEQ ID NO:19, and CDR1, CDR2 and CDR3 domains of the light
chain variable region having the sequence set forth in SEQ ID
NO:21,
[0013] wherein the method comprises at least one administration
cycle, wherein the cycle is a period of eight weeks, wherein for
each of the at least one cycles, four doses of the anti-LAG-3
antibody are administered at a dose of 3, 20, 80, or 240 mg and
four doses of the anti-PD-1 antibody are administered at a dose of
80 or 240 mg. In another embodiment, four doses of the anti-LAG-3
antibody are administered at a dose of about 0.03, 0.25, 1, or 3
mg/kg body weight and four doses of the anti-PD-1 antibody are
administered at a dose of 1 or 3 mg/kg body weight.
[0014] In one embodiment, the anti-LAG-3 antibody and anti-PD-1
antibody are administered at the following doses:
[0015] (a) 3 mg anti-LAG-3 antibody and 80 mg of anti-PD-1
antibody;
[0016] (b) 3 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody;
[0017] (c) 20 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody;
[0018] (d) 80 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody; or
[0019] (e) 240 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody.
[0020] In another embodiment, the anti-LAG-3 antibody and anti-PD-1
antibody are administered at the following doses:
[0021] (a) 0.03 mg/kg anti-LAG-3 antibody and 1 mg/kg of anti-PD-1
antibody;
[0022] (b) 0.03 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody;
[0023] (c) 0.25 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody;
[0024] (d) 1 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody; or
[0025] (e) 3 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody.
[0026] In one embodiment, the dose of the anti-LAG-3 and/or
anti-PD-1 antibody is calculated per mg/kg body weight. In another
embodiment, the dose of the anti-LAG-3 and/or anti-PD-1 antibody is
a flat-fixed dose. In another embodiment, an intermediate dose of
LAG-3 and/or PD-1 is used. For example, LAG-3 could be administered
at 0.4 mg/kg and PD-1 could be administered at 90 mg/kg. In another
embodiment, dosage regimens are adjusted to provide the optimum
desired response (e.g., an effective response).
[0027] In another embodiment, the anti-PD-1 antibody is
administered on Days 1, 15, 29, and 43 of each cycle. In another
embodiment, the anti-LAG-3 antibody is administered on Days 1, 15,
29, and 43 of each cycle. In another embodiment, the anti-PD-1
antibody is administered prior to administration of the anti-LAG-3
antibody. In another embodiment, the anti-PD-1 antibody is
administered after administration of the anti-LAG-3 antibody. In
another embodiment, the treatment consists of up to 12 cycles.
[0028] In one embodiment, the anti-PD-1 antibody and anti-LAG-3
antibody are administered as a first ("front") line of treatment
(e.g., the initial or first treatment). In another embodiment, the
anti-PD-1 antibody and anti-LAG-3 antibody are administered as a
second line of treatment (e.g., after initial treatment with the
same or a different therapeutic, including after relapse and/or
where the first treatment has failed). The anti-LAG-3 and anti-PD-1
antibodies can be administered to a subject by any suitable means.
In one embodiment, the antibodies are formulated for intravenous
administration. In another embodiment, the antibodies are
administered simultaneously (e.g., formulated together in a single
formulation or concurrently as separate formulations).
Alternatively, in another embodiment, the antibodies are
administered sequentially (e.g., as separate formulations). In
another embodiment the anti-LAG-3 antibody is administered within
about 30 minutes (e.g., within about 29, 28, 27, 26, 25, 24, 23,
22, 21, 20, or less minutes) prior to administration of the
anti-PD-1 antibody.
[0029] The efficacy of the treatment methods provided herein can be
assessed using any suitable means. In one embodiment, the treatment
produces at least one therapeutic effect selected from the group
consisting of reduction in size of a tumor, reduction in number of
metastatic lesions over time, complete response, partial response,
and stable disease.
[0030] Also provided are kits that include a pharmaceutical
composition containing an anti-LAG-3 antibody, such as BMS-986016,
and an anti-PD-1 antibody, such as BMS-936558, and a
pharmaceutically-acceptable carrier, in a therapeutically effective
amount adapted for use in the methods described herein. In one
embodiment, the kit comprises:
[0031] (a) a dose of an anti-LAG-3 antibody comprising CDR1, CDR2
and CDR3 domains of the heavy chain variable region having the
sequence set forth in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains
of the light chain variable region having the sequence set forth in
SEQ ID NO:5;
[0032] (b) a dose of an anti-PD-1 antibody comprising CDR1, CDR2
and CDR3 domains of the heavy chain variable region having the
sequence set forth in SEQ ID NO:19, and CDR1, CDR2 and CDR3 domains
of the light chain variable region having the sequence set forth in
SEQ ID NO:21; and
[0033] (c) instructions for using the anti-LAG-3 antibody and
anti-PD-1 antibody in a method of the invention.
[0034] In another aspect, an anti-LAG-3 antibody is provided, the
anti-LAG-3 antibody comprising CDR1, CDR2 and CDR3 domains of the
heavy chain variable region having the sequence set forth in SEQ ID
NO:3, and CDR1, CDR2 and CDR3 domains of the light chain variable
region having the sequence set forth in SEQ ID NO:5, for
co-administration with an anti-PD-1 antibody comprising CDR1, CDR2
and CDR3 domains of the heavy chain variable region having the
sequence set forth in SEQ ID NO:19, and CDR1, CDR2 and CDR3 domains
of the light chain variable region having the sequence set forth in
SEQ ID NO:21, in at least one cycle, wherein for each cycle four
doses of the anti-LAG-3 antibody are administered at a dose of 3,
20, 80, or 240 mg and four doses of the anti-PD-1 antibody are
administered at a dose of 80 or 240 mg. In another embodiment, four
doses of the anti-LAG-3 antibody are administered at a dose of
0.03, 0.25, 1, or 3 mg/kg body weight and four doses of the
anti-PD-1 antibody are administered at a dose of 1 or 3 mg/kg body
weight.
[0035] In another aspect of the invention, the anti-PD-1 antibody
in any of the aforementioned embodiments is replaced by, or
combined with, an anti-PD-L1 or anti-PD-L2 antibody. Accordingly,
the invention also features methods, compositions and kits for
treating tumors in human patients using the above-described
clinically effective dosages of an anti-LAG-3 antibody combined
with the above-described clinically effective dosages of an
anti-PD-1 antibody, wherein the dosage of the PD-1 antibody is
replaced with the same dosage of an anti-PD-L1 or anti-PD-L2
antibody.
BRIEF DESCRIPTION OF THE DRAWINGS
[0036] FIG. 1 shows inhibition of tumor growth in vivo using a
combination treatment of an anti-LAG-3 antibody and an anti-PD-1
antibody in a murine tumor model.
[0037] FIGS. 2A and 2B are schematics illustrating the parts of a
phase I clinical trial.
[0038] FIG. 3 is a schematic illustrating the Screening, Treatment,
Clinical Follow-up, and Survival Follow-up phases of the clinical
trial.
DETAILED DESCRIPTION
I. Definitions
[0039] As used herein, the term "subject" or "patient" is a human
cancer patient (e.g., a patient having an advanced solid tumor,
such as an advanced refractory solid tumor).
[0040] As used herein, "effective treatment" refers to treatment
producing a beneficial effect, e.g., amelioration of at least one
symptom of a disease or disorder. A beneficial effect can take the
form of an improvement over baseline, i.e., an improvement over a
measurement or observation made prior to initiation of therapy
according to the method. A beneficial effect can also take the form
of arresting, slowing, retarding, or stabilizing of a deleterious
progression of a marker of solid tumor. Effective treatment may
refer to alleviation of at least one symptom of a solid tumor. Such
effective treatment may, e.g., reduce patient pain, reduce the size
and/or number of lesions, may reduce or prevent metastasis of a
tumor, and/or may slow tumor growth.
[0041] The term "effective amount" refers to an amount of an agent
that provides the desired biological, therapeutic, and/or
prophylactic result. That result can be reduction, amelioration,
palliation, lessening, delaying, and/or alleviation of one or more
of the signs, symptoms, or causes of a disease, or any other
desired alteration of a biological system. In reference to solid
tumors, an effective amount comprises an amount sufficient to cause
a tumor to shrink and/or to decrease the growth rate of the tumor
(such as to suppress tumor growth) or to prevent or delay other
unwanted cell proliferation. In some embodiments, an effective
amount is an amount sufficient to delay tumor development. In some
embodiments, an effective amount is an amount sufficient to prevent
or delay tumor recurrence. An effective amount can be administered
in one or more administrations. The effective amount of the drug or
composition may: (i) reduce the number of cancer cells; (ii) reduce
tumor size; (iii) inhibit, retard, slow to some extent and may stop
cancer cell infiltration into peripheral organs; (iv) inhibit
(i.e., slow to some extent and may stop tumor metastasis; (v)
inhibit tumor growth; (vi) prevent or delay occurrence and/or
recurrence of tumor; and/or (vii) relieve to some extent one or
more of the symptoms associated with the cancer. In one example, an
"effective amount" is the amount of anti-LAG-3 antibody and the
amount of anti-PD-1 antibody, in combination, clinically proven to
affect a significant decrease in cancer or slowing of progression
of cancer, such as an advanced solid tumor. As used herein, the
terms "fixed dose", "flat dose" and "flat-fixed dose" are used
interchangeably and refer to a dose that is administered to a
patient without regard for the weight or body surface area (BSA) of
the patient. The fixed or flat dose is therefore not provided as a
mg/kg dose, but rather as an absolute amount of the agent (e.g.,
the anti-LAG-3 antibody and/or anti-PD-1 antibody).
[0042] As used herein, a "body surface area (BSA)-based dose"
refers to a dose (e.g., of the anti-LAG-3 antibody and/or anti-PD-1
antibody) that is adjusted to the body-surface area (BSA) of the
individual patient. A BSA-based dose may be provided as mg/kg body
weight. Various calculations have been published to arrive at the
BSA without direct measurement, the most widely used of which is
the Du Bois formula (see Du Bois D, Du Bois E F (June 1916)
Archives of Internal Medicine 17 (6): 863-71; and Verbraecken, J.
et al. (April 2006). Metabolism--Clinical and Experimental 55 (4):
515-24). Other exemplary BSA formulas include the Mosteller formula
(Mosteller R D. N Engl J Med., 1987; 317:1098), the Haycock formula
(Haycock G B, et al., J Pediatr 1978, 93:62-66), the Gehan and
George formula (Gehan E A, George S L, Cancer Chemother Rep 1970,
54:225-235), the Boyd formula (Current, J D (1998), The Internet
Journal of Anesthesiology 2 (2); and Boyd, Edith (1935), University
of Minnesota. The Institute of Child Welfare, Monograph Series, No.
x. London: Oxford University Press), the Fujimoto formula (Fujimoto
S, et al., Nippon Eiseigaku Zasshi 1968; 5:443-50), the Takahira
formula (Fujimoto S, et al., Nippon Eiseigaku Zasshi 1968;
5:443-50), and the Schlich formula (Schlich E, et al., Ernahrungs
Umschau 2010; 57:178-183).
[0043] The term "antibody" describes polypeptides comprising at
least one antibody-derived antigen binding site (e.g., VH/VL region
or Fv, or CDR). Antibodies include known forms of antibodies. For
example, the antibody can be a human antibody, a humanized
antibody, a bispecific antibody, or a chimeric antibody. The
antibody also can be a Fab, Fab'2, ScFv, SMIP, Affibody.RTM.,
nanobody, or a domain antibody. The antibody also can be of any of
the following isotypes: IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2,
IgAsec, IgD, and IgE. The antibody may be a naturally occurring
antibody or may be an antibody that has been altered (e.g., by
mutation, deletion, substitution, conjugation to a non-antibody
moiety). For example, an antibody may include one or more variant
amino acids (compared to a naturally occurring antibody) which
changes a property (e.g., a functional property) of the antibody.
For example, numerous such alterations are known in the art which
affect, e.g., half-life, effector function, and/or immune responses
to the antibody in a patient. The term antibody also includes
artificial polypeptide constructs which comprise at least one
antibody-derived antigen binding site.
[0044] The term "LAG-3" refers to Lymphocyte Activation Gene-3. The
term "LAG-3" includes variants, isoforms, homologs, orthologs and
paralogs. For example, antibodies specific for a human LAG-3
protein may, in certain cases, cross-react with a LAG-3 protein
from a species other than human. In other embodiments, the
antibodies specific for a human LAG-3 protein may be completely
specific for the human LAG-3 protein and may not exhibit species or
other types of cross-reactivity, or may cross-react with LAG-3 from
certain other species, but not all other species (e.g., cross-react
with monkey LAG-3 but not mouse LAG-3). The term "human LAG-3"
refers to human sequence LAG-3, such as the complete amino acid
sequence of human LAG-3 having Genbank Accession No. NP_002277 (SEQ
ID NO:13). The term "mouse LAG-3" refers to mouse sequence LAG-3,
such as the complete amino acid sequence of mouse LAG-3 having
Genbank Accession No. NP_032505. LAG-3 is also known in the art as,
for example, CD223. The human LAG-3 sequence may differ from human
LAG-3 of Genbank Accession No. NP_002277 by having, e.g., conserved
mutations or mutations in non-conserved regions and the LAG-3 has
substantially the same biological function as the human LAG-3 of
Genbank Accession No. NP_002277. For example, a biological function
of human LAG-3 is having an epitope in the extracellular domain of
LAG-3 that is specifically bound by an antibody of the instant
disclosure or a biological function of human LAG-3 is binding to
MHC Class II molecules.
[0045] The term "monkey LAG-3" is intended to encompass LAG-3
proteins expressed by Old World and New World monkeys, including
but not limited to cynomolgus monkey LAG-3 and rhesus monkey LAG-3.
A representative amino acid sequence for monkey LAG-3 is the rhesus
monkey LAG-3 amino acid sequence which is also deposited as Genbank
Accession No. XM_001108923. Another representative amino acid
sequence for monkey LAG-3 is the alternative rhesus monkey sequence
of clone pa23-5 as described in US 2011/0150892 A1. This
alternative rhesus sequence exhibits a single amino acid
difference, at position 419, as compared to the Genbank-deposited
sequence.
[0046] A particular human LAG-3 sequence will generally be at least
90% identical in amino acid sequence to human LAG-3 of Genbank
Accession No. NP_002277 and contains amino acid residues that
identify the amino acid sequence as being human when compared to
LAG-3 amino acid sequences of other species (e.g., murine). In
certain cases, a human LAG-3 can be at least 95%, or even at least
96%, 97%, 98%, or 99% identical in amino acid sequence to LAG-3 of
Genbank Accession No. NP_002277. In certain embodiments, a human
LAG-3 sequence will display no more than 10 amino acid differences
from the LAG-3 sequence of Genbank Accession No. NP_002277. In
certain embodiments, the human LAG-3 can display no more than 5, or
even no more than 4, 3, 2, or 1 amino acid difference from the
LAG-3 sequence of Genbank Accession No. NP_002277. Percent identity
can be determined as described herein.
[0047] As used herein, the terms "Programmed Death 1," "Programmed
Cell Death 1," "Protein PD-1," "PD-1," PD1," "PDCD1," "hPD-1" and
"hPD-I" are used interchangeably, and include variants, isoforms,
species homologs of human PD-1, and analogs having at least one
common epitope with PD-1. The complete PD-1 sequence can be found
under GenBank Accession No. U64863 (SEQ ID NO:29).
[0048] The protein Programmed Death 1 (PD-1) is an inhibitory
member of the CD28 family of receptors, that also includes CD28,
CTLA-4, ICOS and BTLA. PD-1 is expressed on activated B cells, T
cells, and myeloid cells (Agata et al., supra; Okazaki et al.
(2002) Curr. Opin. Immunol. 14: 391779-82; Bennett et al. (2003) J
Immunol 170:711-8). The initial members of the family, CD28 and
ICOS, were discovered by functional effects on augmenting T cell
proliferation following the addition of monoclonal antibodies
(Hutloff et al. Nature (1999); 397:263-266; Hansen et al.
Immunogenics (1980); 10:247-260). PD-1 was discovered through
screening for differential expression in apoptotic cells (Ishida et
al. EMBO J (1992); 11:3887-95). The other members of the family,
CTLA-4 and BTLA, were discovered through screening for differential
expression in cytotoxic T lymphocytes and TH1 cells, respectively.
CD28, ICOS and CTLA-4 all have an unpaired cysteine residue
allowing for homodimerization. In contrast, PD-1 is suggested to
exist as a monomer, lacking the unpaired cysteine residue
characteristic in other CD28 family members.
[0049] The PD-1 gene is a 55 kDa type I transmembrane protein that
is part of the Ig gene superfamily (Agata et al. (1996) Int Immunol
8:765-72). PD-1 contains a membrane proximal immunoreceptor
tyrosine inhibitory motif (ITIM) and a membrane distal
tyrosine-based switch motif (ITSM) (Thomas, M. L. (1995) J Exp Med
181:1953-6; Vivier, E and Daeron, M (1997) Immunol Today
18:286-91). Although structurally similar to CTLA-4, PD-1 lacks the
MYPPPY motif that is critical for B7-1 and B7-2 binding. Two
ligands for PD-1 have been identified, PD-L1 and PD-L2, that have
been shown to downregulate T cell activation upon binding to PD-1
(Freeman et al. (2000) J Exp Med 192:1027-34; Latchman et al.
(2001) Nat Immunol 2:261-8; Carter et al. (2002) Eur J Immunol
32:634-43). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1,
but do not bind to other CD28 family members. PD-L1 is abundant in
a variety of human cancers (Dong et al. (2002) Nat. Med. 8:787-9).
The interaction between PD-1 and PD-L1 results in a decrease in
tumor infiltrating lymphocytes, a decrease in T-cell receptor
mediated proliferation, and immune evasion by the cancerous cells
(Dong et al. (2003) J. Mol. Med. 81:281-7; Blank et al. (2005)
Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin.
Cancer Res. 10:5094-100). Immune suppression can be reversed by
inhibiting the local interaction of PD-1 with PD-L1, and the effect
is additive when the interaction of PD-1 with PD-L2 is blocked as
well (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99:12293-7;
Brown et al. (2003) J. Immunol. 170:1257-66).
[0050] Consistent with PD-1 being an inhibitory member of the CD28
family, PD-1 deficient animals develop various autoimmune
phenotypes, including autoimmune cardiomyopathy and a lupus-like
syndrome with arthritis and nephritis (Nishimura et al. (1999)
Immunity 11:141-51; Nishimura et al. (2001) Science 291:319-22).
Additionally, PD-1 has been found to play a role in autoimmune
encephalomyelitis, systemic lupus erythematosus, graft-versus-host
disease (GVHD), type I diabetes, and rheumatoid arthritis (Salama
et al. (2003) J Exp Med 198:71-78; Prokunina and Alarcon-Riquelme
(2004) Hum Mol Genet 13:R143; Nielsen et al. (2004) Lupus 13:510).
In a murine B cell tumor line, the ITSM of PD-1 was shown to be
essential to block BCR-mediated Ca.sup.2+-flux and tyrosine
phosphorylation of downstream effector molecules (Okazaki et al.
(2001) PNAS 98:13866-71).
[0051] "Programmed Death Ligand-1 (PD-L1)" is one of two cell
surface glycoprotein ligands for PD-1 (the other being PD-L2) that
downregulate T cell activation and cytokine secretion upon binding
to PD-1. The term "PD-L1" as used herein includes human PD-L1
(hPD-L1), variants, isoforms, and species homologs of hPD-L1, and 5
analogs having at least one common epitope with hPD-L1. The
complete hPD-L1 sequence can be found under GenBank Accession No.
Q9NZQ7.
IIa. Anti-LAG-3 Antibodies
[0052] Anti-human-LAG-3 antibodies (or VH/VL domains derived
therefrom) suitable for use in the invention can be generated using
methods well known in the art. Alternatively, art recognized
anti-LAG-3 antibodies can be used. For example, the anti-human
LAG-3 antibody described in US2011/0150892 A1, the teachings of
which are hereby incorporated by reference, and referred to as
monoclonal antibody 25F7 (also known as "25F7" and "LAG3.1) can be
used. Other art recognized anti-LAG-3 antibodies that can be used
include IMP731 described in US 2011/007023, the teachings of which
also are hereby incorporated by reference.
[0053] Antibodies that compete with any of the above-referenced
art-recognized antibodies for binding to LAG-3 also can be
used.
[0054] An exemplary anti-LAG-3 antibody is BMS-986016 comprising
heavy and light chains comprising the sequences shown in SEQ ID
NOs:1 and 2, respectively, or antigen binding fragments and
variants thereof, as described in PCT/US13/48999, the teachings of
which are hereby incorporated by reference.
[0055] In other embodiments, the antibody has the heavy and light
chain CDRs or variable regions of BMS-986016. Accordingly, in one
embodiment, the antibody comprises CDR1, CDR2, and CDR3 domains of
the VH region of BMS-986016 having the sequence set forth in SEQ ID
NO:3, and CDR1, CDR2 and CDR3 domains of the VL region of
BMS-986016 having the sequence set forth in SEQ ID NO:5. In another
embodiment, the antibody comprises CDR1, CDR2 and CDR3 domains
comprising the sequences set forth in SEQ ID NOs:7, 8, and 9,
respectively, and CDR1, CDR2 and CDR3 domains comprising the
sequences set forth in SEQ ID NOs:10, 11, and 12, respectively. In
another embodiment, the antibody comprises VH and/or VL regions
comprising the amino acid sequences set forth in SEQ ID NO:3 and/or
SEQ ID NO: 5, respectively. In another embodiment, the antibody
comprises heavy chain variable (VH) and/or light chain variable
(VL) regions encoded by the nucleic acid sequences set forth in SEQ
ID NO:4 and/or SEQ ID NO:6, respectively. In another embodiment,
the antibody competes for binding with and/or binds to the same
epitope on LAG-3 as the above-mentioned antibodies. In another
embodiment, the antibody binds an epitope of human LAG-3 comprising
the amino acid sequence PGHPLAPG (SEQ ID NO:14). In another
embodiment, the antibody binds an epitope of human LAG-3 comprising
the amino acid sequence HPAAPSSW (SEQ ID NO:15) or PAAPSSWG (SEQ ID
NO:16).
[0056] In another embodiment, the antibody has at least about 90%
variable region amino acid sequence identity with the
above-mentioned antibodies (e.g., at least about 90%, 95% or 99%
variable region identity with SEQ ID NO:3 or SEQ ID NO:5).
IIb. Anti-PD-1 Antibodies
[0057] Anti-human-PD-1 antibodies (or VH and/or VL domains derived
therefrom) suitable for use in the invention can be generated using
methods well known in the art. Alternatively, art recognized
anti-PD-1 antibodies can be used. For example, monoclonal
antibodies 5C4 (referred to herein as Nivolumab or BMS-936558),
17D8, 2D3, 4H1, 4A11, 7D3, and 5F4, described in WO 2006/121168,
the teachings of which are hereby incorporated by reference, can be
used. Other known PD-1 antibodies include Lambrolizumab (MK-3475)
described in WO 2008/156712, and AMP-514 described in WO
2012/145493, the teachings of which are hereby incorporated by
reference. Further known PD-1 antibodies and other PD-1 inhibitors
include those described in WO 2009/014708, WO 03/099196, WO
2009/114335 and WO 2011/161699, the teachings of which are hereby
incorporated by reference. Antibodies that compete with any of
these art-recognized antibodies or inhibitors for binding to PD-1
also can be used.
[0058] An exemplary anti-PD-1 antibody is BMS-936558 comprising
heavy and light chains comprising the sequences shown in SEQ ID
NOs:17 and 18, respectively, or antigen binding fragments and
variants thereof.
[0059] In other embodiments, the antibody has heavy and light chain
CDRs or variable regions of BMS-936558. Accordingly, in one
embodiment, the antibody comprises CDR1, CDR2, and CDR3 domains of
the VH of BMS-936558 having the sequence set forth in SEQ ID NO:19,
and CDR1, CDR2 and CDR3 domains of the VL of BMS-936558 having the
sequence set forth in SEQ ID NO:21. In another embodiment, the
antibody comprises CDR1, CDR2 and CDR3 domains comprising the
sequences set forth in SEQ ID NOs:23, 24, and 25, respectively, and
CDR1, CDR2 and CDR3 domains comprising the sequences set forth in
SEQ ID NOs:26, 27, and 28, respectively. In another embodiment, the
antibody comprises VH and/or VL regions comprising the amino acid
sequences set forth in SEQ ID NO: 19 and/or SEQ ID NO: 21,
respectively. In another embodiment, the antibody comprises heavy
chain variable (VH) and/or light chain variable (VL) regions
encoded by the nucleic acid sequences set forth in SEQ ID NO:20
and/or SEQ ID NO:22, respectively. In another embodiment, the
antibody competes for binding with and/or binds to the same epitope
on PD-1 as the above-mentioned antibodies. In another embodiment,
the antibody has at least about 90% variable region amino acid
sequence identity with the above-mentioned antibodies (e.g., at
least about 90%, 95% or 99% variable region identity with SEQ ID
NO:19 or SEQ ID NO:21).
IIc. Anti-PD-L1 Antibodies
[0060] Anti-human-PD-L1 antibodies (or VH and/or VL domains derived
therefrom) suitable for use in the invention can be generated using
methods well known in the art. Alternatively, art recognized
anti-PD-L1 antibodies can be used. For example, human anti-PD-L1
antibodies disclosed in U.S. Pat. No. 7,943,743, the contents of
which are hereby incorporated by reference, can be used. Such
anti-PD-L1 antibodies include 3G10, 12A4 (also referred to as
BMS-936559), 10A5, 5F8, 10H10, 1B12, 7H1, 11E6, 12B7, and 13G4.
Other art recognized anti-PD-L1 antibodies which can be used
include those described in, for example, U.S. Pat. Nos. 7,635,757
and 8,217,149, U.S. Publication No. 2009/0317368, and PCT
Publication Nos. WO 2011/066389 and WO 2012/145493, the teachings
of which also are hereby incorporated by reference. Antibodies that
compete with any of these art-recognized antibodies or inhibitors
for binding to PD-L1 also can be used.
III. Pharmaceutical Compositions
[0061] Pharmaceutical compositions suitable for administration to
human patients are typically formulated for parenteral
administration, e.g., in a liquid carrier, or suitable for
reconstitution into liquid solution or suspension for intravenous
administration.
[0062] In general, such compositions typically comprise a
pharmaceutically acceptable carrier. As used herein, the term
"pharmaceutically acceptable" means approved by a government
regulatory agency or listed in the U.S. Pharmacopeia or another
generally recognized pharmacopeia for use in animals, particularly
in humans. The term "carrier" refers to a diluent, adjuvant,
excipient, or vehicle with which the compound is administered. Such
pharmaceutical carriers can be sterile liquids, such as water and
oils, including those of petroleum, animal, vegetable or synthetic
origin, such as peanut oil, soybean oil, mineral oil, sesame oil,
glycerol polyethylene glycol ricinoleate, and the like. Water or
aqueous solution saline and aqueous dextrose and glycerol solutions
may be employed as carriers, particularly for injectable solutions
(e.g., comprising an anti-LAG-3 or anti-PD-1 antibody). Liquid
compositions for parenteral administration can be formulated for
administration by injection or continuous infusion. Routes of
administration by injection or infusion include intravenous,
intraperitoneal, intramuscular, intrathecal and subcutaneous. In
one embodiment, the anti-LAG-3 and/or anti-PD-1 antibodies are
administered intravenously (e.g., in separate formulations or
together (in the same formulation or in separate
formulations)).
IV. Patient Populations
[0063] Provided herein are clinical methods for treating solid
tumors cancer (e.g., advanced refractory solid tumors) in human
patients using a combination of an anti-LAG-3 antibody and an
anti-PD-1 antibody.
[0064] Examples of cancers that may be treated using the methods of
the invention, include liver cancer, bone cancer, pancreatic
cancer, skin cancer, cancer of the head or neck, breast cancer,
lung cancer, cutaneous or intraocular malignant melanoma, renal
cancer, uterine cancer, ovarian cancer, colorectal cancer, colon
cancer, rectal cancer, cancer of the anal region, stomach cancer,
testicular 20 cancer, uterine cancer, carcinoma of the fallopian
tubes, carcinoma of the endometrium, carcinoma of the cervix,
carcinoma of the vagina, carcinoma of the vulva, non-Hodgkin's
lymphoma, cancer of the esophagus, cancer of the small intestine,
cancer of the endocrine system, cancer of the thyroid gland, cancer
of the parathyroid gland, cancer of the adrenal gland, sarcoma of
soft tissue, cancer of the urethra, cancer of the penis, solid
tumors of 25 childhood, lymphocytic lymphoma, cancer of the
bladder, cancer of the kidney or ureter, carcinoma of the renal
pelvis, neoplasm of the central nervous system (CNS), primary CNS
lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma,
pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous
cell cancer, environmentally induced cancers including those
induced by asbestos, hematologic malignancies 30 including, for
example, multiple myeloma, B-cell lymphoma, Hodgkin
lymphoma/primary mediastinal B-cell lymphoma, non-Hodgkin's
lymphomas, acute myeloid lymphoma, chronic myelogenous leukemia,
chronic lymphoid leukemia, follicular lymphoma, diffuse large
B-cell lymphoma, Burkitt's lymphoma, immunoblastic large cell
lymphoma, precursor B-lymphoblastic lymphoma, mantle cell lymphoma,
acute lymphoblastic leukemia, mycosis fungoides, anaplastic large
cell lymphoma, T-cell lymphoma, and precursor T-lymphoblastic
lymphoma, and any combinations of said cancers. The present
invention is also applicable to treatment of metastatic
cancers.
[0065] In one embodiment, the human patient suffers from non-small
cell lung cancer (NSCLC) or a virally-related cancer (e.g., a human
papilloma virus (HPV)-related tumor) or gastric adenocarcinoma. In
a particular embodiment, the HPV-related tumor is HPV+ head and
neck cancer (HNC). In another particular embodiment, the gastric
adenocarcinoma is associated with Epstein-Barr virus (EBV)
infection.
[0066] Patients can be tested or selected for one or more of the
above described clinical attributes prior to, during or after
treatment.
V. Combination Therapy
[0067] Combination therapies provided herein involve administration
of an anti-LAG-3 antibody and another antibody that blocks an
inhibitory immune receptor (e.g., a receptor, which upon binding to
its natural ligand, inhibits/neutralizes activity, such as
cytotoxic activity), particularly an anti-PD-1 antibody, to treat
subjects having solid tumors (e.g., advanced refractory solid
tumors).
[0068] In one embodiment, the invention provides an anti-LAG-3
antibody and an anti-PD-1 antibody in combination according to a
defined clinical dosage regimen, to treat subjects having a solid
tumor (e.g., an advanced refractory solid tumor). In a particular
embodiment, the anti-LAG-3 antibody is BMS-986016. In another
embodiment, the anti-PD-1 antibody is BMS-936558. In another
embodiment, dosage regimens are adjusted to provide the optimum
desired response (e.g., an effective response).
[0069] As used herein, adjunctive or combined administration
(coadministration) includes simultaneous administration of the
compounds in the same or different dosage form, or separate
administration of the compounds (e.g., sequential administration).
Thus, the anti-LAG-3 and anti-PD-1 antibodies can be simultaneously
administered in a single formulation. Alternatively, the anti-LAG-3
and anti-PD-1 antibodies can be formulated for separate
administration and are administered concurrently or sequentially
(e.g., one antibody is administered within about 30 minutes prior
to administration of the second antibody).
[0070] For example, the anti-PD1 antibody can be administered first
followed by (e.g., immediately followed by) the administration of
the anti-LAG-3 antibody, or vice versa. In one embodiment, the
anti-PD-1 antibody is administered prior to administration of the
anti-LAG-3 antibody. In another embodiment, the anti-PD-1 antibody
is administered after administration of the anti-LAG-3 antibody. In
another embodiment, the anti-LAG-3 antibody and anti-PD-1 antibody
are administered concurrently. Such concurrent or sequential
administration preferably results in both antibodies being
simultaneously present in treated patients.
VI. Treatment Protocols
[0071] Suitable treatment protocols for treating a solid tumor in a
human patient include, for example, administering to the patient an
effective amount of each of:
[0072] (a) an anti-LAG-3 antibody comprising CDR1, CDR2 and CDR3
domains of the heavy chain variable region having the sequence set
forth in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains of the light
chain variable region having the sequence set forth in SEQ ID
NO:5,
[0073] (b) an anti-PD-1 antibody comprising CDR1, CDR2 and CDR3
domains of the heavy chain variable region having the sequence set
forth in SEQ ID NO:19, and CDR1, CDR2 and CDR3 domains of the light
chain variable region having the sequence set forth in SEQ ID
NO:21,
[0074] wherein the method comprises at least one administration
cycle, wherein the cycle is a period of eight weeks, wherein for
each of the at least one cycles, at least four doses of the
anti-LAG-3 antibody are administered at a flat dose of about 1, 3,
10, 20, 50, 80, 100, 130, 150, 180, 200, 240 or 280 mg and at least
four doses of the anti-PD-1 antibody are administered at flat dose
of about 50, 80, 100, 130, 150, 180, 200, 240 or 280 mg. In another
embodiment, four doses of the anti-LAG-3 antibody are administered
at a dose of 0.01, 0.03, 0.25, 0.1, 0.3, 1 or 3, 5, 8 or 10 mg/kg
body weight and four doses of the anti-PD-1 antibody are
administered at a dose of 0.1, 0.3, 1, 3, 5, 8 or 10 mg/kg body
weight.
[0075] In one embodiment, the anti-LAG-3 antibody and anti-PD-1
antibody are administered at the following doses:
[0076] (a) 3 mg anti-LAG-3 antibody and 80 mg of anti-PD-1
antibody;
[0077] (b) 3 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody;
[0078] (c) 20 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody;
[0079] (d) 80 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody; or
[0080] (e) 240 mg anti-LAG-3 antibody and 240 mg of anti-PD-1
antibody.
[0081] In another embodiment, the anti-LAG-3 antibody and anti-PD-1
antibody are administered at the following doses:
[0082] (a) 0.3 mg/kg anti-LAG-3 antibody and 1 mg/kg of anti-PD-1
antibody;
[0083] (b) 0.3 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody;
[0084] (c) 0.25 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody;
[0085] (d) 1 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody; or
[0086] (e) 3 mg/kg anti-LAG-3 antibody and 3 mg/kg of anti-PD-1
antibody.
[0087] In one embodiment, the dose of the anti-LAG-3 and/or
anti-PD-1 antibody is calculated per body weight, e.g., mg/kg body
weight. In another embodiment, the dose of the anti-LAG-3 and/or
anti-PD-1 antibody is a flat-fixed dose. In another embodiment, the
dose of the anti-LAG-3 and/or anti-PD-1 antibody is varied over
time. For example, the anti-LAG-3 antibody and/or anti-PD-1
antibody may be initially administered at a high dose and may be
lowered over time. In another embodiment, the anti-LAG-3 antibody
and/or anti-PD-1 antibody is initially administered at a low dose
and increased over time.
[0088] In another embodiment, the amount of the anti-LAG-3 and/or
anti-PD-1 antibodies administered is constant for each dose. In
another embodiment, the amount of antibody administered varies with
each dose. For example, the maintenance (or follow-on) dose of the
antibody can be higher or the same as the loading dose which is
first administered. In another embodiment, the maintenance dose of
the antibody can be lower or the same as the loading dose.
[0089] In another embodiment, the anti-LAG-3 and/or anti-PD-1
antibodies are formulated for intravenous administration. In one
embodiment, the anti-PD-1 antibody is administered on Days 1, 15,
29, and 43 of each cycle. In another embodiment, the anti-LAG-3
antibody is administered on Days 1, 15, 29, and 43 of each
cycle.
[0090] In other embodiments, the anti-LAG-3 and/or anti-PD-1
antibodies are administered once per week, once every or three two
weeks, once per month or as long as a clinical benefit is observed
or until there is a complete response, confirmed progressive
disease or unmanageable toxicity.
[0091] In another embodiment, a cycle of administration is eight
weeks, which can be repeated, as necessary. In another embodiment,
the treatment consists of up to 12 cycles.
[0092] In another embodiment, 4 doses of the anti-PD-1 antibody are
administered per eight week cycle. In another embodiment, 4 doses
of the anti-LAG-3 antibody are administered per eight week
cycle.
[0093] In another embodiment, the anti-PD-1 antibody and anti-LAG-3
antibody are administered as a first line of treatment (e.g., the
initial or first treatment). In another embodiment, the anti-PD-1
antibody and anti-LAG-3 antibody are administered as a second line
of treatment (e.g., after the initial or first treatment, including
after relapse and/or where the first treatment has failed).
[0094] In another aspect, the invention features any of the
aforementioned embodiments, wherein the anti-PD-1 antibody is
replaced by, or combined with, an anti-PD-L1 or anti-PD-L2
antibody.
VII. Outcomes
[0095] With respect to target lesions, responses to therapy may
include:
TABLE-US-00001 Complete Response (CR) Disappearance of all target
lesions. Any (RECIST V1.1) pathological lymph nodes (whether target
or non-target) must have reduction in short axis to <10 mm.
Partial Response (PR) At least a 30% decrease in the sum of the
(RECIST V1.1) diameters of target lesions, taking as reference the
baseline sum diameters. Progressive Disease (PD) At least a 20%
increase in the sum of the (RECIST V1.1) diameters of target
lesions, taking as reference the smallest sum on study (this
includes the baseline sum if that is the smallest on study). In
addition to the relative increase of 20%, the sum must also
demonstrate an absolute increase of at least 5 mm. (Note: the
appearance of one or more new lesions is also considered
progression). Stable Disease (SD) Neither sufficient shrinkage to
qualify for (RECIST V1.1) PR nor sufficient increase to qualify for
PD, taking as reference the smallest sum diameters while on study.
Immune-related Complete Disappearance of all target lesions. Any
Response (irCR) pathological lymph nodes (whether target (irRECIST)
or non-target) must have reduction in short axis to <10 mm.
Immune-related Partial At least a 30% decrease in the sum of
Response (irPR) (irRECIST) diameters of target lesions and all new
measurable lesions (ie Percentage Change in Tumor Burden), taking
as reference the baseline sum diameters. Note: the appearance of
new measurable lesions is factored into the overall Tumor Burden,
but does not automatically qualify as progressive disease until the
sum of the diameters increases by 20% when compared to nadir.
Immune-related At least a 20% increase in Tumor Burden Progressive
Disease (irPD) (ie the sum of diameters of target lesions,
(irRECIST) and any new measurable lesions) taking as reference the
smallest sum on study (this includes the baseline sum if that is
the smallest on study). In addition to the relative increase of
20%, the sum must also demonstrate an absolute increase of at least
5 mm. Tumor assessments using immune-related criteria for
progressive disease incorporates the contribution of new measurable
lesions. Each net percentage change in tumor burden per assessment
accounts for the size and growth kinetics of both old and new
lesions as they appear. Immune-related Stable Neither sufficient
shrinkage to qualify for Disease (irSD) (irRECIST) irPR nor
sufficient increase to qualify for irPD, taking as reference the
smallest sum diameters while on study.
[0096] With respect to non-target lesions, responses to therapy may
include:
TABLE-US-00002 Complete Response (CR) Disappearance of all
non-target lesions. (RECIST V1.1) All lymph nodes must be non-
pathological in size (<10 mm short axis). Non-CR/Non-PD
Persistence of one or more non-target (RECIST V1.1) lesion(s).
Progressive Disease (PD) Unequivocal progression of existing non-
(RECIST V1.1) target lesions. The appearance of one or more new
lesions is also considered progression. Immune-related
Disappearance of all non-target lesions. Complete Response (irCR)
All (irRECIST) lymph nodes must be non-pathological in size (<10
mm short axis). Immune-related Increases in number or size of
non-target Progressive Disease (irPD) lesion(s) does not constitute
progressive (irRECIST) disease unless/until Tumor Burden increases
by 20% (ie the sum of the diameters at nadir of target lesions and
any new measurable lesions increases by the required amount).
Non-target lesions are not considered in the definition of Stable
Disease and Partial Response.
[0097] Patients treated according to the methods disclosed herein
preferably experience improvement in at least one sign of cancer.
In one embodiment, improvement is measured by a reduction in the
quantity and/or size of measurable tumor lesions. In another
embodiment, lesions can be measured on chest x-rays or CT or MRI
films. In another embodiment, cytology or histology can be used to
evaluate responsiveness to a therapy.
[0098] In one embodiment, the patient treated exhibits a complete
response (CR), a partial response (PR), stable disease (SD),
immune-related complete disease (irCR), immune-related partial
response (irPR), or immune-related stable disease (irSD). In
another embodiment, the patient treated experiences tumor shrinkage
and/or decrease in growth rate, i.e., suppression of tumor growth.
In another embodiment, unwanted cell proliferation is reduced or
inhibited. In yet another embodiment, one or more of the following
can occur: the number of cancer cells can be reduced; tumor size
can be reduced; cancer cell infiltration into peripheral organs can
be inhibited, retarded, slowed, or stopped; tumor metastasis can be
slowed or inhibited; tumor growth can be inhibited; recurrence of
tumor can be prevented or delayed; one or more of the symptoms
associated with cancer can be relieved to some extent.
[0099] In other embodiments, administration of effective amounts of
the anti-LAG-3 antibody and anti-PD-1 antibody according to any of
the methods provided herein produces at least one therapeutic
effect selected from the group consisting of reduction in size of a
tumor, reduction in number of metastatic lesions appearing over
time, complete remission, partial remission, or stable disease. In
still other embodiments, the methods of treatment produce a
comparable clinical benefit rate (CBR=CR+PR+SD.gtoreq.6 months)
better than that achieved by an anti-LAG-3 antibody or anti-PD-1
antibody alone. In other embodiments, the improvement of clinical
benefit rate is about 20% 20%, 30%, 40%, 50%, 60%, 70%, 80% or more
compared to an anti-LAG-3 antibody or anti-PD-1 antibody alone.
VIII. Kits and Unit Dosage Forms
[0100] Also provided herein are kits which include a pharmaceutical
composition containing an anti-LAG-3 antibody, such as BMS-986016,
and an anti-PD-1 antibody, such as BMS-936558, and a
pharmaceutically-acceptable carrier, in a therapeutically effective
amount adapted for use in the preceding methods. The kits
optionally also can include instructions, e.g., comprising
administration schedules, to allow a practitioner (e.g., a
physician, nurse, or patient) to administer the composition
contained therein to administer the composition to a patient having
cancer (e.g., a solid tumor). The kit also can include a
syringe.
[0101] Optionally, the kits include multiple packages of the
single-dose pharmaceutical compositions each containing an
effective amount of the anti-LAG-3 or anti-PD-1 antibody for a
single administration in accordance with the methods provided
above. Instruments or devices necessary for administering the
pharmaceutical composition(s) also may be included in the kits. For
instance, a kit may provide one or more pre-filled syringes
containing an amount of the anti-LAG-3 or anti-PD-1 antibody.
[0102] In one embodiment, the present invention provides a kit for
treating a solid tumor in a human patient, the kit comprising:
[0103] (a) a dose of an anti-LAG-3 antibody comprising CDR1, CDR2
and CDR3 domains of the heavy chain variable region having the
sequence set forth in SEQ ID NO:3, and CDR1, CDR2 and CDR3 domains
of the light chain variable region having the sequence set forth in
SEQ ID NO:5;
[0104] (b) a dose of an anti-PD-1 antibody comprising CDR1, CDR2
and CDR3 domains of the heavy chain variable region having the
sequence set forth in SEQ ID NO:19, and CDR1, CDR2 and CDR3 domains
of the light chain variable region having the sequence set forth in
SEQ ID NO:21; and
[0105] (c) instructions for using the anti-LAG-3 antibody and
anti-PD-1 antibody in the methods described herein.
[0106] The following examples are merely illustrative and should
not be construed as limiting the scope of this disclosure in any
way as many variations and equivalents will become apparent to
those skilled in the art upon reading the present disclosure.
[0107] The contents of all references, GenBank entries, patents and
published patent applications cited throughout this application are
expressly incorporated herein by reference.
EXAMPLES
Example 1: Pre-Clinical Pharmacology of Anti-PD-1 Antibody
(BMS-936558)
[0108] BMS-936558 is a fully human, IgG4 (kappa) isotype monoclonal
antibody that binds to PD-1 with nanomolar affinity as measured by
surface plasmon resonance using a Biacore.RTM. biosensor system and
a high degree of specificity, thus precluding binding to its
ligands PD-L1 and PD-L2. BMS-936558 does not bind other related
family members, such as BTLA, CTLA-4, ICOS or CD28. Pre-clinical
testing of BMS-936558 demonstrated that binding to PD-1 results in
enhanced T cell proliferation and release of interferon-gamma
(IFN-gamma) in vitro. The heavy and light chain amino acid
sequences of BMS-936558 are provided in SEQ ID NOs: 1 and 2,
respectively.
Example 2: In Vivo Toxicity of Anti-PD-1 Antibody Novolumab
(BMS-936558)
[0109] Toxicology studies in cynomolgus monkeys confirmed that
BMS-936558 was well tolerated at doses up to 50 mg/kg given twice
weekly for 27 doses. Drug-related findings were limited to a
reversible decrease in triiodothyronine (T3) by 28%, without
concomitant abnormalities in other markers of thyroid function
(data not shown).
Example 3: Clinical Pharmacology and Safety of Anti-PD-1 Antibody
(BMS-936558)
[0110] The overall safety experience with BMS-936558, as a
monotherapy or in combination with other therapeutics, is based on
experience in approximately 1500 subjects treated to date. In
general for monotherapy, the safety profile is similar across tumor
types. The one exception is pulmonary inflammation adverse events
(AEs) which may be numerically greater in subjects with NSCLC
because in some cases, it can be difficult to distinguish between
BMS-936558-related and unrelated causes of pulmonary symptoms and
radiographic changes. The safety profile is generally consistent
across completed and ongoing clinical trials with no maximum
tolerated dose reached at any dose tested up to 10 mg/kg. There was
no pattern in the incidence, severity, or causality of adverse
events to BMS-936558 dose level.
[0111] Study CA209003 has contributed to most of the clinical
experience with BMS-936558 in subjects with NSCLC and other solid
malignancies to date. CA209003 was a Phase 1 multi-dose escalation
study in subjects with previously treated advanced or metastatic
melanoma, RCC, NSCLC, colorectal cancer, or hormone-refractory
prostate cancer. In CA209003, subjects were administered BMS-936558
intravenously every 2 weeks with doses of 0.1, 0.3, 1, 3, or 10
mg/kg. No maximum tolerated dose was identified in CA209003. The
maximum dose level evaluated was 10 mg/kg. The incidence, severity,
and relationship of adverse events were generally similar across
dose levels and tumor types.
[0112] As of 3 Jul. 2012, 296 (97.4%) out of 304 subjects treated
with BMS-936558 had at least 1 reported adverse event regardless of
causality. There was no pattern in the incidence, severity, or
relationship of adverse events to the BMS-936558 dose level.
Treatment-related adverse events of any grade occurred in 220
(72.4%) of subjects. The most frequent drug-related adverse events
occurring in >5% of subjects included fatigue (25.7%), rash
(13.5%), diarrhea (11.8%), pruritus (10.2%), nausea (7.9%),
decreased appetite (7.9%), hemoglobin decreased (5.9%), and pyrexia
(5.3%). Most treatment-related adverse events were low grade (Grade
1 or 2). Treatment-related high-grade (Grade 3 or 4) adverse events
were reported in 45 (14.8%) of subjects, with the most common being
fatigue (1.6%), decreased appetite (1.0%), and diarrhea (1.0%). At
least one serious adverse event (SAE) was reported for 150 (49.3%)
of the 304 subjects at all dose levels. Grade 3-4 SAEs were
reported for 23 subjects (7.6%). Drug-related SAEs of any grade
occurred in 11.5% of subjects. Grade 3-4 drug-related SAEs reported
in at least 2 subjects included diarrhea (3 subjects [1.0%]),
pneumonitis (3 subjects [1.0%]), pneumonia (2 subjects [0.7%]), and
lipase increased (2 subjects [0.7%]). Similar to the overall
adverse event profile, there was no apparent relationship in the
incidence or severity of drug-related adverse events to BMS-936558
dose level. There were no apparent differences in the frequency of
adverse events based on subjects' tumor type.
[0113] Selected treatment-related adverse events have occurred with
low frequency (<5%), but are considered clinically meaningful
because they require greater vigilance for early recognition and
prompt intervention. These adverse events include alanine
aminotransferase (ALT) increased (4.3%), aspartate aminotransferase
(AST) increased (3.6%), pneumonitis (3.3%), hypothyroidism (3.0%),
hyperthyroidism (1.3%), adrenal insufficiency (0.7%), and colitis
(0.7%). Grade 3-4 events of pneumonitis were reported in 3 subjects
(1.0%) as described above (1 event was Grade 4). Grade 3 events of
colitis, ALT increased, and AST increased were reported in 2
subjects (0.7%) each. Grade 3 events of adrenal insufficiency,
hyperthyroidism, and hypothyroidism were reported in 1 subject
(0.3%) each. Because of the potential for clinically meaningful
BMS-936558-related adverse events requiring early recognition and
prompt intervention, management algorithms have been developed for
suspected pulmonary toxicity, diarrhea or suspected colitis,
hepatotoxicity, endocrinopathy, and nephrotoxicity.
[0114] Treatment-related adverse events leading to discontinuation
were reported in 18 (5.9%) of the 304 treated subjects on CA209003.
The only events reported in more than 1 subject were pneumonitis (4
subjects [1.3%]) and hepatitis (2 subjects [0.7%]). There were 3
(1.0%) drug-related deaths; each occurred after the development of
pneumonitis.
[0115] The safety of BMS-936558 in combination with other
therapeutics is being explored in several ongoing clinical
trials.
Example 4: Pharmacokinetics of Anti-PD-1 Antibody (BMS-936558)
[0116] Single-dose pharmacokinetics (PK) of BMS-936558 were
evaluated in 39 subjects with multiple tumor types in CA209001 in
the dose range of 0.3 to 10 mg/kg. The median Tmax across dose
levels ranged from 1.6 to 3.1 hours with individual values ranging
from 0.9 to 7 hours. The PK of BMS-936558 was linear in the range
of 0.3 to 10 mg/kg with dose-proportional increase in Cmax and
AUC(INF) and low to moderate intersubject variability was observed
at each dose level (i.e., coefficient of variation [CV] ranging
from 16 to 45%). Geometric mean clearance (CLT) after a single IV
dose ranged from 0.13 to 0.19 mL/h/kg, while mean volume of
distribution (Vz) varied between 83 to 113 mL/kg across doses. The
mean terminal T-HALF of BMS-936558 was 17 to 25 days, consistent
with half-life of endogenous IgG4, indicating that the elimination
mechanism of BMS-936558 may be similar to IgG4. Both elimination
and distribution of BMS-936558 appeared to be independent of dose
within the dose range studied. In a multiple dose study of multiple
tumor types (CA209003), available data from 128 subjects, mean
T-HALF was 21-24 hours and median T-max ranged from 0.6 to 3.3
across dose levels, which aligns with the single dose data.
Example 5: Phase I Clinical Trial with Anti-PD-1 Antibody
(BMS-936558)
[0117] BMS-936558 has demonstrated clinical activity in a completed
Phase 1 single-dose study and 2 ongoing multiple-dose escalation
studies (Phase 1 monotherapy: CA209003 and Phase 1b combination
therapy with ipilimumab) in subjects with NSCLC, melanoma, RCC, and
other malignancies. Tumor response was determined by modified
Response Evaluation Criteria in Solid Tumors (RECIST) established
by the NCI. The evaluable population consists of 294 subjects with
a variety of solid tumor malignancies (melanoma, n=138; NSCLC,
n=122; RCC, n=34) who are currently being treated with
nivolumab.
[0118] In CA209003, an objective response rate (ORR) of 31.1% (33
of 106 response-evaluable subjects) was reported in subjects with
melanoma treated with BMS-936558 monotherapy every 2 weeks (Q2W) at
doses ranging from 0.1 to 10 mg/kg. The majority of responses were
durable and exceeded 6 months.
[0119] In the most active dose range (3 to 10 mg/kg), an ORR of
13.5% to 27.8% was reported among subjects with NSCLC with a
24-week progression-free survival rate (PFSR) of 23% to 51%.
Durable responses were observed in both squamous and non-squamous
subtypes.
[0120] Of the 34 response-evaluable RCC subjects in CA209003,
responses were reported in both the 1-mg/kg (5 of 18 subjects,
27.8%) and 10-mg/kg (5 of 16 subjects, 31.3%) treatment groups.
Estimated progression-free survival rate (PFSR) at 24 weeks was 50%
in the 1-mg/kg and 67% in the 10-mg/kg BMS-936558 treatment
groups.
[0121] Preliminary results from the Phase 1b study of combination
therapy with BMS-936558 and ipilimumab suggest an advantage in
combining two T cell-targeted therapies for subjects with melanoma.
In the 0.3 mg/kg BMS-936558+3 mg/kg ipilimumab treatment group,
responses were observed in 5 out of 14 evaluable subjects (35.7%, 1
complete response and 2 partial responses by conventional modified
World Health Organization [mWHO] criteria, and 2 partial responses
by immune-related mWHO criteria). In the 1 mg/kg BMS-936558+3 mg/kg
ipilimumab treatment group, responses were observed in 9 out of 15
evaluable subjects (60%, 3 CRs and 6 PRs; all by conventional mWHO
criteria). In the 3 mg/kg BMS-936558+3 mg/kg ipilimumab treatment
group, objective responses were observed in 4 out of 6 evaluable
subjects (66.7%, 3 partial responses by conventional mWHO criteria
and 1 partial response by immune-related mWHO criteria). Further
details are provided by Wolchok et al. (2013) NEJM 369(2):122-33,
and/or PCT/US2013/040764.
Example 6: Pre-Clinical Pharmacology of Anti-LAG-3 Antibody
(BMS-986016)
[0122] BMS-986016 is a fully human antibody specific for human
LAG-3 that was isolated from immunized transgenic mice expressing
human immunoglobulin genes. It is expressed as an IgG4 isotype
antibody that includes a stabilizing hinge mutation (S228P) for
attenuated Fc receptor binding in order to reduce or eliminate the
possibility of antibody- or complement-mediated target cell
killing. The heavy and light chain amino acid sequences of
BMS-986016 are provided in SEQ ID NOs:17 and 18, respectively.
[0123] The ability of BMS-986016 to bind recombinant human LAG-3
antigen was determined using Biacore and enzyme-linked
immunosorbent assay (ELISA). Binding to human and primate LAG-3+
transfectants and to activated human or primate T cells was
measured using flow cytometric and Scatchard analyses. BMS-986016
binds to human LAG-3 with high affinity (K.sub.D=0.12-0.5 nM), and
inhibits the binding of LAG-3 to cells expressing its ligand, MHC
class II (IC50, 0.67 nM). BMS-986016 binds to cynomolgus LAG-3 on
transfected CHO cells and on activated cynomolgus T cells with a
lower affinity (EC50, 21.5-34.3 nM) than to activated human T
cells. A high concentration of BMS-986016, in the absence of
secondary co-stimulation, elicits no measurable cytokine response
from cultured human peripheral blood cells nor does the drug
mediate measurable antibody-dependent or complement-dependent
killing of target cells. BMS-986016 promotes the activation of an
antigen-specific mouse T cell hybridoma expressing human LAG-3 in
co-culture with an MHC class II-positive antigen-presenting cell.
In addition, BMS-986016 enhances activation of human T cells in
superantigen stimulation assays when added alone or in combination
with BMS-936558 (anti-PD-1 antibody).
Example 7: Toxicity of Anti-LAG-3 Antibody (BMS-986016) Alone or in
Combination with Anti-PD-1 Antibody (BMS-936558)
[0124] The following preclinical toxicology studies were
performed:
[0125] A. Four-Week Intermittent (QW) Intravenous Exploratory
Combination Pharmacodynamic and Toxicity Study in Cynomolgus
Monkeys with Anti-LAG3.1 Antibody (a Precursor of BMS-986016) and
BMS-936558
[0126] The key results were as follows. Anti-LAG3.1 administered at
50 mg/kg/week, alone or in combination with 50 mg/kg/week
BMS-936558, did not result in any adverse changes.
No-observed-adverse-effect level (NOAEL) for single-agent
anti-LAG3.1 was considered to be 50 mg/kg/week (AUC[0-168h]=231,000
.mu.gh/mL), and NOAEL for anti-LAG3.1 in combination with 50
mg/kg/week BMS-936558 was considered to be 50 mg/kg/week (mean
anti-LAG3.1 AUC[0-168h]=210,000 .mu.gh/mL; mean
BMS-936558AUC[0-168h]=159,500 .mu.gh/mL).
[0127] B. GLP-Compliant Four-Week Intravenous Combination Toxicity
Study in Cynomolgus Monkeys with a 6-Week Recovery with BMS-986016
and BMS-936558
[0128] The key results were as follows. Single-agent BMS-986016
administered at up to 100 mg/kg/week did not result in adverse
changes. Single-agent BMS-936558 administered at 50 mg/kg/week
resulted in slight to minimal non-reversible lymphoplamacytic
inflammation of the choroid plexus of the brain, which was
considered non-adverse given the lower severity and incidence of
the lymphoplamacytic inflammation compared to combination treatment
with BMS-986016 and BMS-936558, lack of vasculitis or tissue
destruction, and absence of clinical manifestations during the
course of treatment. Combined administration of BMS-986016 and
BMS-936558 (100 and 50 mg/kg/week, respectively) resulted in
morbidity of 1 male out of 9 monkeys on study Day 29. From Days 26
to 29, this monkey presented with elevated body temperature,
shivers, red or clear nasal discharge, fecal changes (unformed,
scant or absent feces), decreased feeding behavior, mild
dehydration, sneezing, decreased activity, and hunched posture.
After 2 days of veterinary care and antibiotic treatment, this
animal did not show any improvement and was euthanized on Day 29
for poor clinical condition.
[0129] There were no remarkable gross necropsy findings.
Histopathological findings in this monkey included: slight
lymphoplamacytic inflammation of the choroid plexus; minimal to
moderate lymphohistocytic inflammation of the vasculature of the
brain parenchyma, meninges, spinal cord (cervical and lumbar); and
minimal to moderate mixed cell inflammation of the epididymes,
seminal vesicles and testes. Clinical pathology changes indicated
decreases in red blood cell count, hemoglobin concentration and
hematocrit whose cause was unclear, and an increase in fibrinogen
correlating with the inflammation observed in the central nervous
system (CNS) and male reproductive tract.
[0130] Additional histopathological findings upon combination
administration of BMS-986016 and BMS-936558 (100 and 50 mg/kg/week,
respectively) were limited to minimal to slight non-reversible
lymphoplamacytic inflammation of the choroid plexus in the brain in
7 of 8 remaining monkeys, and minimal lymphohistiocytic
inflammation of the vasculature of the brain parenchyma in 1 of 8
remaining monkeys, whose reversibility could not be assessed.
[0131] NOAEL for single-agent BMS-986016 was considered to be 100
mg/kg/week (mean AUC[0-168h]=474,000 .mu.gh/mL); NOAEL for
single-agent BMS-936558 was considered to be 50 mg/kg/week (mean
AUC[0-168h]=193,000 .mu.gh/mL); NOAEL for combination of BMS-986016
and BMS-936558 was not determined. However, the combination therapy
was generally well tolerated and clinical signs of toxicity were
observed in only 1 of 9 monkeys (approximately 10%). Therefore,
100/50 mg/kg/week BMS-986016/nivolumab (mean BMS-986016
AUC[0-168h]=514,000 .mu.gh/mL; mean nivolumab AUC[0-168h]=182,000
.mu.gh/mL) was considered the STD10.
[0132] The doses administered (100 mg/kg BMS-986016 and 50 mg/kg
BMS-936558) are .gtoreq.10 times higher than the maximum doses
proposed for the current study. The starting dose of 20 mg (0.25
mg/kg) for BMS-986016 monotherapy (Part A) is less than 1/10 of the
human equivalent of the cynomolgus monkey NOAEL (636 mg; 8.0
mg/kg), and is below the HED after a linear adjustment of the NOAEL
target exposure for the highest affinity difference estimate of
265-fold (24 mg; 0.30 mg/kg). The calculated safety multiple for
exposures at the 20 mg (0.25 mg/kg) starting dose is 315-fold based
on the cynomolgus monkey NOAEL of 100 mg/kg/week without accounting
for affinity differences.
[0133] The starting dose of 3 mg (0.03 mg/kg) for BMS-986016 for
the combination therapy (Part B) is based on a linear adjustment of
the cynomolgus monkey STD10 for the 265-fold highest affinity
difference estimate with an added 10-fold safety factor. The
maximum recommended starting dose (MRSD) for BMS-986016 based on a
100 mg/kg/week STD10 is 0.03 mg/kg in humans. The starting dose of
80 mg (1 mg/kg) for BMS-936558 for the combination therapy (Part B)
is based on known human BMS-936558 PK parameters with an added
10-fold safety factor. The MRSD for BMS-936558 based on the 50
mg/kg/week cynomolgus monkey STD10 is 4.3 mg/kg in humans, and has
been further reduced to identify a dose with acceptable levels of
adverse events.
[0134] C. GLP-Compliant Tissue Cross Reactivity Study in Human and
Select Cynomolgus Monkey Tissues with BMS-986016.
[0135] Positive staining with BMS-986016-FITC was observed in the
plasma membrane or plasma membrane granules following human
tissues: mononuclear leukocytes of the urinary bladder, blood
cells, colon--large intestine, eye, esophagus, small intestine,
stomach, kidney, lung, lymph node, placenta, salivary gland, skin,
spleen, thymus, tonsil, uterus--cervix, and uterus--endometrium;
and hematopoetic cells of the bone marrow. In addition, staining
with BMS-986016-FITC was observed in the cytoplasm of the human
pituitary endocrine cell epithelium. Within the limited panel of
cynomolgus monkey tissues evaluated, staining with BMS-986016-FITC
was observed in the plasma membrane or plasma membrane granules of
the mononuclear leukocytes of the spleen. With scientific reports
of LAG-3-expressing cells in germinal centers and interfollicular
T-cell areas of normal human lymphoid tissues (lymph node, tonsil,
spleen, thymus, bone marrow and mucosal-associated lymphoid tissue)
and having the morphology and distribution of lymphocytes, the
staining of mononuclear leukocytes and hematopoietic cells with
BMS-986016-FITC in this study (in the human and cynomolgus monkey
tissues) was anticipated. Given that LAG-3 mRNA is expressed in the
human pituitary and LAG3.1-G4P-FITC staining was observed in
adenohypophysis of the human pituitary in a pilot tissue cross
reactivity study, BMS-986016-FITC staining of human pituitary
endocrine cell epithelium cytoplasm and cytoplasmic granules was
also anticipated. Although BMS-986016 is not expected to have
access to the cytoplasmic compartment in vivo and the repeat-dose
toxicology studies in monkeys showed no effects on the pituitary
gland, these findings may be of clinical significance.
[0136] D. In Vitro Cytokine Release and Lymphocyte Activation
Assessment with BMS-986016 Using Human Peripheral Blood Mononuclear
Cells.
[0137] BMS-986016 did not induce cytokine release when presented to
human PBMCs regardless of concentration, donor, or incubation time.
The levels of cytokines observed were either at or near the assay
lower limits of quantification with no evidence of dose-dependence
or pattern across donors (IL-1.beta., IL-2, IL-5, IL-10, IL-12p70,
and IFN-.gamma.) or were generally overlapping with cytokine levels
from PBMCs incubated with negative controls (IL-6, IL-8,
TNF-.alpha.).
[0138] Consistent with the lack of cytokine release, there was no
evidence that BMS-986016 induced T or NK cell activation, as
measured by surface expression of CD25 and CD69. Expression levels
of these markers on T and NK cells following stimulation with
BMS-986016 were similar to those observed upon stimulation with
negative controls.
[0139] Overall, these data indicate that BMS-986016 does not
possess agonistic potential to induce either T or NK cellular
activation or cytokine release.
Example 8: Preclinical Pharmacokinetics of Anti-LAG-3 Antibody
(BMS-986016)
[0140] In accordance with regulatory guidelines for
biotechnology-derived pharmaceuticals (ICH Harmonised Tripartite
Guideline, S6(R1) Preclinical Safety Evaluation of
Biotechnology-Derived Pharmaceuticals. International Conference on
Harmonisation, 2011), no metabolism studies with BMS-986016 have
been conducted in animals. The expected in vivo degradation of
monoclonal antibodies (mAbs) is to small peptides and amino acids
via biochemical pathways that are independent of cytochrome P450
enzymes.
[0141] BMS-986016 demonstrated favorable pharmacokinetic (PK)
properties in cynomolgus monkeys. From both single-dose and
repeat-dose IV PK studies, BMS-986016 decayed bi-exponentially and
the exposure was approximately dose-proportional. The systemic
clearance (CLTp) ranges from 0.12 to 0.22 mL/h/kg and a terminal
half-life (T-HALF) 133 to 414 hours. The volume of distribution at
steady state (Vss) was 62 to 72 mL/kg, suggesting limited
distribution outside the plasma. Anti-BMS-986016 antibodies were
detected in some monkeys but the presence of anti-BMS-986016
antibodies appeared to have no impact on BMS-986016 exposure.
Example 9: Inhibition of Tumor Growth In Vivo by Combination
Treatment with Anti-LAG-3 Antibody and Anti-PD-1 Antibody
[0142] An experiment was conducted in a murine tumor model to test
the hypothesis that the combination of anti-LAG-3 and anti-PD-1
would potentiate anti-tumor efficacy. These studies evaluated tumor
growth inhibition in syngeneic tumor models (Sa1N fibrosarcoma and
MC38 colon adenocarcinoma) and monitored acceleration of
autoimmunity in the non-obese diabetic (NOD) model. Anti-LAG-3
antibody administration resulted in both overall tumor growth
inhibition and an increase in the number of tumor-free (TF) mice in
those treatment groups, as shown in FIG. 1. Anti-LAG-3 antibody
administered in combination with anti-PD-1 antibody provided
enhanced anti-tumor activity above the activity of either agent
alone. For example, in multiple Sa1N tumor models, anti-LAG-3
antibody resulted in 20%-30% TF mice compared to control and
anti-PD-1 antibody-treated mice (0%-10% TF mice), while the
combination of anti-LAG-3 and anti-PD-1 antibodies resulted in
60%-90% TF mice. In the MC38 model, anti-LAG-3 antibody showed
modest tumor growth inhibition alone, but when administered in
combination with anti-PD-1 antibody, resulted in enhanced antitumor
activity above that observed for anti-PD-1 antibody alone (80% vs.
40% TF mice, respectively).
Example 10: Phase 1 Trial in Patients Having Solid Tumors
[0143] A phase 1 trial of anti-LAG-3 antibody (BMS-986016) and
anti-PD-1 antibody (BMS-936558) is conducted in patients having
advanced solid tumors to demonstrate the efficacy of administering
BMS-986016 and BMS-936558 as a combination treatment.
[0144] 1. Objectives
[0145] The primary objective of the study is to assess the safety
and tolerability of BMS-986016 given in combination with BMS-936558
and to identify dose limiting toxicities (DLTs) and the maximally
tolerated dose (MTD) of the combination, in subjects with advanced
solid tumors.
[0146] Secondary objectives include assessing the preliminary
anti-tumor activity of the combination of BMS-986016 and BMS-936558
in subjects with advanced solid tumors, characterizing the
pharmacokinetics (PK) of BMS-986016 and BMS-936558 when
co-administered, monitoring immunogenicity of BMS-986016 and
BMS-936558 administered as combination therapy, and assessing the
effect of BMS-986016 and BMS-936558 on corrected QT ("QTc").
Additional exploratory objectives include assessing the
pharmacodynamic effects of BMS-986016 and BMS-936558 combination
therapy based on select biomarkers in the peripheral blood and
tumor biopsy specimens, characterizing T cell function during
BMS-986016 and BMS-936558 combination therapy, assessing the 2-year
landmark overall survival in subjects treated with BMS-986016 and
BMS-936558, exploring preliminary antitumor activity of BMS-986016
and BMS-936558 combination therapy in subjects with advanced solid
tumors, characterizing pharmacokinetics and exposure-response
relationships in subjects treated with BMS-986016 and BMS-936558,
and investigating the relationship between clinical efficacy and
selected peripheral and tumor biomarkers.
[0147] 2. Study Design and Duration
[0148] This is a Phase 1, open-label study of BMS-986016
administered as a single agent and in combination with BMS-936558
(nivolumab) to subjects with advanced solid tumors. The study is
conducted in 3 parts. Part A and Part B consist of a 3+3+3 dose
escalation design with BMS-986016 administered as a single agent
(Part A) or in combination with BMS-936558 (Part B) in subjects
with advanced solid tumors. Treatment in Part B is initiated upon
the decision to escalate to the third dose cohort in Part A (in
accordance with dose escalation rules). Subsequently, escalation in
the 2 parts proceeds in parallel. At no point does the dose of
BMS-986016 administered in combination with BMS-936558 (Part B)
exceed doses of BMS-986016 that have been demonstrated previously
to be safe on the monotherapy dose escalation arm (Part A). Part C
consists of cohort expansion in 6 disease-restricted populations of
approximately 16 subjects each, with BMS-986016 administered in
combination with BMS-936558. Treatment in Part C is initiated when
the maximum tolerated dose (MTD) (or maximum administered dose
(MAD) if no MTD is determined) for Part B has been determined. The
doses selected for Part C do not exceed the Part B MTD or MAD, but
dose determination may incorporate assessment of other data,
including toxicities and PK and pharmacodynamic data from Parts A
and B. A schematic of the study is provided in FIG. 2.
[0149] Subjects complete up to 4 periods of the study as follows:
Screening (up to 28 days), Treatment (up to a maximum of twelve
8-week cycles of study therapy), Clinical Follow-up (135 days), and
Survival Follow-up (up to 2 years following the first dose of study
drug; a longer follow-up period could be considered in selected
cases if an efficacy signal is apparent). During this period,
diagnostic imaging may be performed every 12 weeks until
progression in subjects who discontinue due to CR, and in subjects
with PR at the end of Cycle 12.
[0150] The Treatment Period consists of up to twelve 8-week
treatment cycles. Each treatment cycle is comprised of 4 doses of
either BMS-986016 alone (Part A) or in combination with BMS-936558
(Parts B and C), administered on Days 1, 15, 29, and 43 of each
treatment cycle. In Parts B and C when both antibodies are
administered in combination, nivolumab will be given first followed
by BMS-986016 within 30 minutes of completing the infusion of
nivolumab. Tumor response is evaluated using RECIST v1.1. Subjects
are allowed to continue study therapy until the occurrence of
either: (1) confirmed complete response (CR), (2) completion of the
maximum number of twelve 8-week cycles, (3) progressive disease
(PD), (4) clinical deterioration, and/or (5) meeting other criteria
for discontinuation. Treatment beyond progression is allowed in
select subjects with initial RECIST v1.1-defined PD who are
receiving clinical benefit and tolerating treatment. Subjects who
discontinue treatment enter a 135-day Clinical Follow-up
period.
[0151] After completion of the Clinical Follow-up period, subjects
enter the Survival Follow-up period. During this period, clinic
visits or telephone contact every 12 weeks are performed to assess
survival status. The duration of this period is up to 2 years
following the first dose of study drug, although a longer follow-up
period is considered in selected cases if an efficacy signal is
apparent. Subjects in the Survival Follow-up period who have
disease progression are allowed to receive tumor-directed therapy
as required. A study schematic is depicted in FIG. 3.
[0152] Assessments, including physical examinations, vital sign
measurements, 12-lead ECG, and clinical laboratory evaluations are
performed at selected times throughout the dosing interval.
Subjects are closely monitored for adverse events throughout the
study. Blood samples are collected for up to 4 hours following the
start of study drug administration for pharmacokinetic
analysis.
[0153] Subjects are allowed to continue on therapy for up to twelve
8-week cycles or until confirmed complete response, progressive
disease, clinical deterioration, or meeting criteria for
discontinuation. Subjects may be on study for a total of up to
approximately 2.3 years, including a 28-day screening period, up to
twelve 8-week cycles of treatment, a 135-day clinical follow-up
period, and up to 2 years of follow-up for survival (beginning from
the first dose of study drug). The total duration of the study is
expected to be approximately 5 years from the time of the first
visit of the first subject to the required survival follow-up of
the last subject enrolled.
[0154] 3. Dose Escalation
[0155] Part A
[0156] In Part A, a 3+3+3 design is used to assess the safety of
BMS-986016 given as single agent. A fourth subject may be enrolled
at the beginning of a dose escalation cohort, if subject is able to
start the first day of dosing within approximately one week of the
third subject in the same dose escalation cohort. The dosages
during dose escalation are provided in FIGS. 2A and 2B and Table 1
(set forth below). Three subjects (or 4, if applicable) are
initially treated in each dose cohort. In Dose Cohort 1, each of
the first 3 subjects (or 4, if applicable) is designated as
sentinel subjects and begin a treatment at least 5 days apart.
Subjects in subsequent cohorts are not required to observe the
5-day interval between treatment start dates. Dose escalation in
Part A proceeds as follows: [0157] If 0 of the first 3 subjects (or
4, if applicable) experience a dose-limiting toxicity (DLT) within
the DLT evaluation interval, a new cohort of 3 subjects (or 4, if
applicable) is treated at the next higher dose level. [0158] If 1
of 3 subjects (or 4, if applicable) experience a DLT within the DLT
evaluation interval, that cohort is expanded to 6 subjects. [0159]
If 2 of 6 subjects experience a DLT within the DLT evaluation
interval, that cohort is expanded to 9 subjects. [0160] If
.gtoreq.2 of 3 (or 4, if applicable), .gtoreq.3 of 6, or .gtoreq.3
of 9 subjects experience DLTs within a dose cohort during the DLT
evaluation interval, then that dose level is determined to have
exceeded the MTD.
TABLE-US-00003 [0160] TABLE 1 Dose Escalation Schedule for Part A -
BMS-986016 Monotherapy BMS-986016 Dose Dose Cohort Number Total
Subjects (IV; mg) 1 n = approximately 3-9 20 2 n = approximately
3-9 80 3 n = approximately 3-9 240 4 n = approximately 3-9 800
Total N = approximately 12-36
[0161] Prior to declaring the MTD (or MAD), any cohort previously
established to be safe is expanded to obtain additional experience
or to investigate dose levels intermediate to those defined in the
protocol. Dose escalation rules (cohort size, DLT evaluation
interval, cohort expansion criteria, etc.) apply to these expanded
or additional cohorts. A maximum of 9 subjects are enrolled in any
additional or expanded dose cohorts.
[0162] No within-subject dose escalations are permitted. If a dose
level is found to exceed the MTD, subjects enrolled in that dose
level are treated at a lower dose.
[0163] Part B
[0164] Treatment in Part B is initiated after the decision is made
to escalate to the third dose cohort in Part A (in accordance with
dose escalation rules). Subsequently, escalation in the 2 parts
proceeds in parallel. At no point does the dose of BMS-986016
administered in combination with BMS-936558 (Part B) exceed doses
of BMS-986016 that have been demonstrated previously to be safe on
the monotherapy dose escalation arm (Part A). Treatment assignments
for subjects eligible for both Part A and Part B alternates between
the 2 parts, with consecutively treated subjects assigned to
different parts through interactive voice response system (IVRS)
whenever possible. If there are no openings available in the part
to which the subject is assigned by this algorithm, the subject is
assigned to the next open cohort or part. As in Part A, a 3+3+3
design is also be used in Part B to assess the safety of BMS-986016
given in combination with nivolumab. A fourth subject may be
enrolled at the beginning of a dose escalation cohort, if subject
is able to start the first day of dosing within approximately one
week of the third subject in the same dose escalation cohort. The
dosages evaluated during dose escalation are provided in FIGS. 2A
and 2B and Table B (set forth below). As in Part A, each of the
first 3 subjects (or 4, if applicable) in the first dose cohort in
Part B will be designated as sentinel subjects and will begin
treatment at least 5 days apart.
TABLE-US-00004 TABLE 2 Dose Escalation Schedule for Part B -
BMS-986016 in Combination with BMS-936558 BMS-986016 BMS-936558
Dose Cohort Dose Dose Number Total Subjects (IV; mg) (IV; mg) 1 n =
approximately 3-9 3 80 2 n = approximately 3-9 3 240 3 n =
approximately 3-9 20 240 4 n = approximately 3-9 80 240 5 n =
approximately 3-9 240 240 Total N = approximately 15-45
[0165] Three subjects are treated initially in each dose cohort. In
Dose Cohort 1, each of the first 3 subjects, designated as sentinel
subjects, begins treatment at least 5 days apart. Subjects in
subsequent cohorts are not be required to observe the 5-day
interval between treatment start dates.
[0166] Dose escalation in Part B proceeds as described for Part A.
If the MTD is exceeded in Dose Cohort 2, the subsequent cohort is
treated with 20 mg of BMS-986016 and 80 mg of BMS-936558. If this
dose combination is found to be safe, escalation proceeds at the
previously defined BMS-986016 doses, maintaining the BMS-936558
dose at 80 mg.
[0167] If no MTD is reached through Dose Cohort 5, then additional
cohorts of BMS-986016 given in combination with BMS-936558 are
considered based on the aggregate safety experience during dose
escalation.
[0168] Prior to declaring the MTD (or MAD), any cohort previously
established to be safe is expanded in order to obtain additional
experience or to investigate dose levels intermediate to those
defined in the protocol. Dose escalation rules (cohort size, DLT
evaluation interval, cohort expansion criteria, etc.) apply to
these expanded or additional cohorts. A maximum of 9 subjects are
enrolled in any additional or expanded dose cohorts.
[0169] No within-subject dose escalations are permitted. If a dose
level is found to exceed the MTD, subjects enrolled in that dose
level are reduced to a lower dose.
[0170] 4. Cohort Expansion
[0171] The purpose of cohort expansion is to gather additional
safety, tolerability, preliminary efficacy, pharmacokinetic, and
pharmacodynamic information regarding the combination of BMS-986016
and BMS-936558. The doses selected for Part C do not exceed the MTD
(or MAD if no MTD is determined) in Part B, but may incorporate
assessment of other data including toxicities and PK and
pharmacodynamic data from Parts A and B. Doses include doses
intermediate to those evaluated in Part B. Modeling is used to help
inform the selection of the combination dose level to carry forward
in Part C if a dose below the MTD is chosen. Six expansion cohorts
are restricted to the tumor types listed below in Tables 3A and 3B.
Continuous evaluation of toxicity events in the cohort expansions
is performed throughout enrollment in the expansion cohorts. If, at
any time, the aggregate rate of treatment-related toxicities
meeting DLT criteria exceeds 33% across all subjects treated in the
Part C cohort expansion, further enrollment is interrupted.
Depending on the nature and grade of the toxicity and after
assessing the risk:benefit ratio, a new dose(s) for all cohorts is
initiated at a previously tested lower dose level or at a dose
level intermediate to previously tested lower dose levels.
[0172] Upon determination of the MTD (or MAD if no MTD is
determined) in Part A, a BMS-986016 monotherapy cohort is evaluated
in cohort expansion. This expansion cohort is restricted to the
tumor type(s) found to be responsive to BMS-986016 monotherapy. The
dose selected for monotherapy expansion does not exceed the Part A
MTD (or MAD if no MTD is determined) and incorporates assessment of
other data, including toxicities and PK and pharmacodynamic data
from Part A. The dose selected is intermediate to those tested in
Part A. Modeling is used to help inform the selection of the dose
level to carry forward in Part C if a dose below the MTD is
chosen.
TABLE-US-00005 TABLE 3A Tumor Types Eligible For Part C - Cohort
Expansion Combination Therapy Tumor Type Total Subjects Melanoma:
naive to ICMARs.sup.a approximately 16 Melanoma: anti-PD-1 or
anti-PD-L1 antibody approximately 16 as most recent therapy.sup.b
NSCLC.sup.c: naive to ICMARs.sup.a approximately 16 NSCLC.sup.c:
anti-PD-1 or anti-PD-L1 antibody as approximately 16 most recent
therapy.sup.b HPV.sup.d-positive head and neck cancer naive to
approximately 16 ICMARs.sup.a Gastric adenocarcinoma naive to
ICMARs.sup.a approximately 16 Total approximately 96 .sup.aICMARs:
immune cell-modulating antibody regimens (such as, but not limited
to, ipilimumab, tremelimumab, anti-PD-1, anti-PD-L1, anti-PD-L2,
anti-KIR, anti-CD137, and/or anti-OX40 antibodies) .sup.bSubjects
with anti-PD-1 or anti-PD-L1 antibody as most recent therapy are
nonresponsive subjects with progression within 16 weeks of starting
therapy. Subjects must provide informed consent within 60 days
after the last dose of anti-PD-1 or anti-PD-L1 antibody and should
not have discontinued antibody therapy due to serious and/or
life-threatening toxicity (e.g., dose-limiting toxicity in prior
study). Subjects with anti-PD-1 or anti-PD-L1 antibody as most
recent therapy cannot have had prior exposure to any other ICMARs.
.sup.cNSCLC: non-small cell lung cancer .sup.dHPV: human papilloma
virus
TABLE-US-00006 TABLE 3B Tumor Types Eligible For Part C - Cohort
Expansion Combination Therapy Tumor Type Total Subjects Melanoma:
naive to ICMARs.sup.e approximately 16 Melanoma: prior anti-CTLA-4
and anti-PD-1 or approximately 16 anti-PD-L1 antibody therapy.sup.f
NSCLC.sup.g: naive to ICMARs.sup.a approximately 16 NSCLC.sup.c:
anti-PD-1 or anti-PD-L1 antibody as approximately 16 most recent
therapy.sup.b Head and neck cancer naive to ICMARs.sup.a
approximately 16 Gastric adenocarcinoma naive to ICMARs.sup.a
approximately 16 Total approximately 96 .sup.aICMARs: immune
cell-modulating antibody regimens (such as, but not limited to,
anti-CTLA-4, and anti-PD-1 or anti-PD-L1, anti-PD-L2, anti-KIR,
anti-CD137, and/or anti-OX40 antibodies) .sup.bMelanoma subjects
progressing while-on or after receiving anti-CTLA-4 and anti-PD-1
or anti-PD-L1 antibody therapies (in sequential or combination
regimens), are eligible. Non-eligible melanoma subjects in this
group include those with: 1) last does of anti-CTLA-4 antibody
therapy received within 100 days of first dose of study medication;
2) prior exposure to ICMARs other than anti-CTLA-4, anti-PD-1 or
anti-PD-L1 antibody therapy regimens; 3) discontinuation from
anti-CTLA-4, anti-PD-1 or anti-PD-L1 antibody therapy due to
serious and/or life-threatening toxicity (e.g., dose-limiting
toxicity in prior exposure). .sup.cNSCLC: non-small cell lung
cancer .sup.dNSCLC subjects whose disease progresses while-on or
after therapy with anti-PD-1 or anti-PD-L1 antibody as most recent
therapy. Subject should not have discontinued antibody therapy due
to serious and/or life-threatening toxicity (e.g., dose-limiting
toxicity in prior study). Subjects with anti-PD-1 or anti-PD-L1
antibody as most recent therapy cannot have had prior exposure to
any other ICMARs.
[0173] 5. Dose Limiting Toxicities
[0174] BMS-986016 has the potential to augment the frequency and
severity of previously described adverse events associated with
BMS-936558, or to produce new toxicities. For the purpose of
guiding decisions regarding dose escalation in Part A and Part B,
dose-limiting toxicity (DLT) is determined based on the incidence,
intensity, and duration of adverse events that are related to study
drug and that occur within 56 days (8 weeks) of initiation of study
drug (i.e., the DLT evaluation interval, through the completion of
Cycle 1). The severity of adverse events is graded according to
National Cancer Institute (NCI) Common Terminology Criteria for
Adverse Events (CTCAE) v4.0. For the purpose of subject management,
DLTs that occur at any time, whether during dose escalation (Part A
and Part B) or cohort expansion (Part C) result in all study
drug(s) being held, pending evaluation of the event's relatedness
to study drug. Subjects must meet criteria for re-treatment prior
to re-initiation of study treatment.
[0175] Subjects who withdraw from the study during the DLT
evaluation interval for reasons other than a DLT may be replaced at
the same dose level. In the case that an infusion cannot be
administered at a scheduled visit during the DLT evaluation
interval, it must be administered as soon as possible. If the delay
is between 1 and 7 days, the procedures at the original scheduled
visit should be performed and subjects will be considered evaluable
for DLT determination. If the delay is more than 7 days, the dose
will be considered missed and will not be replaced. For the purpose
of making decisions on dose escalation from a safety perspective,
subjects will be considered evaluable if they have received 3 out
of the 4 scheduled BMS-986016 doses in Part A (or 3 out of 4
schedules BMS-986016 and nivolumab doses in Part B) through the 8
week observation period, only if the one missed dose was secondary
to progressive disease or non-medical reasons. Unevaluable subjects
may be replaced at the same dose level.
[0176] Hepatic, non-hematologic, and hematologic DLTs are defined
separately as outlined below.
[0177] Hepatic DLT
[0178] Any of the following drug-related events are considered a
hepatic DLT: [0179] ALT or AST>8.times.ULN, regardless of
duration, or [0180] ALT or AST>5.times. and .ltoreq.8.times.ULN,
that fails to return to .ltoreq.Grade 1 within 2 weeks despite
medical intervention, or [0181] Total bilirubin >5.times.ULN, or
[0182] ALT or AST>3.times.ULN and concurrent total bilirubin
>2.times.ULN
[0183] Non-Hematologic DLT
[0184] Any of the following drug-related events are considered a
non-hematologic DLT: [0185] Grade 2 immune related-eye pain or
reduction in visual acuity that requires systemic treatment, or
[0186] Grade 2 eye pain or reduction in visual acuity that does not
respond to topical therapy and that does not improve to Grade 1
within 2 weeks of initiation of topical therapy, or [0187]
.gtoreq.Grade 3 non-hepatic or non-hematologic toxicity with the
exceptions noted below.
[0188] The following Grade 3 non-hematologic events are not
considered DLTs: [0189] Grade 3 electrolyte abnormality that lasts
<72 hours, is not clinically complicated, and resolves
spontaneously or responds to conventional medical intervention
[0190] Grade 3 increase in amylase or lipase that persists for
<3 weeks and is not associated with clinical or radiographic
evidence of pancreatitis [0191] Grade 3 nausea or vomiting that
lasts <48 hours, and resolves to .ltoreq.Grade 1 either
spontaneously or with conventional medical intervention [0192]
Grade 3 fever that lasts <72 hours, and is not associated with
hemodynamic compromise (including hypotension, or clinical or
laboratory evidence of end organ perfusion impairment) [0193] Grade
3 endocrinopathy that is well controlled by hormone replacement
[0194] Grade 3 tumor flare (defined as pain, irritation, or rash
that localizes to sites of known or suspected tumor) [0195] Grade 3
fatigue for less than 7 days
[0196] Hematologic DLT
[0197] Any of the following drug-related events are considered a
hematologic DLT: [0198] Grade 4 febrile neutropenia of any duration
[0199] Grade 4 neutropenia that lasts >5 days [0200] Grade 4
thrombocytopenia [0201] Grade 4 anemia [0202] Grade 3
thrombocytopenia associated with clinically significant bleeding
[0203] Grade 3 febrile neutropenia that lasts >48 hours [0204]
Grade 3 hemolysis
[0205] 6. Inclusion Criteria
[0206] Signed Written Informed Consent
[0207] The subject must sign and date the IRB/IEC approved written
informed consent form prior to the performance of any study-related
procedures that are not considered part of standard of care.
[0208] Consent for tumor biopsy samples for Part A or Part B Dose
Escalation: Subject must consent to allow the acquisition of
existing formalin-fixed paraffin-embedded (FFPE) tumor tissue,
either a block or unstained slides, for performance of correlative
studies. If an archived sample is not available, subject must
consent to allow a pre-treatment tumor biopsy. In either case,
study personnel must ensure that the tissue block or slides
physically exist prior to initiating therapy. Subjects unable to
provide an archived tumor sample and who either do not consent to a
pre-treatment tumor biopsy or do not have accessible lesions are
not eligible. (However, subjects whose pre-treatment biopsy yields
inadequate tissue quantity or quality are not ineligible on this
basis alone.)
[0209] Consent for tumor biopsy samples for Part C Cohort
Expansion: Subjects with melanoma or head and neck tumors: All
subjects in the 2 melanoma cohorts and all subjects in the head and
neck tumor cohort are required to undergo pre-treatment and
on-treatment biopsies; therefore, subjects must have a lesion
located such that the specimen can be obtained at acceptable
clinical risk as judged by the Investigator. Biopsy sites for any
subjects must be distinct from evaluable lesions. Subjects in the
melanoma and head and neck cancer cohorts who do not meet these
criteria are not eligible; however, subjects whose screening biopsy
yields inadequate tissue quantity or quality are not be ineligible
on this basis alone. Subjects in the remaining cohorts (NSCLC or
gastric adenocarcinoma): Subject must consent to allow the
acquisition of existing formalin-fixed paraffin-embedded (FFPE)
tumor tissue, either a block or unstained slides, for performance
of correlative studies. If an archived sample is not available,
subject must consent to allow a pre-treatment tumor biopsy. In
either case, study personnel must ensure that the tissue block or
slides physically exist prior to initiating therapy. Subjects
unable to provide an archived tumor sample and who either do not
consent to a pre-treatment tumor biopsy or do not have accessible
lesions are not eligible. (However, subjects whose pre-treatment
biopsy yields inadequate tissue quantity or quality are not
ineligible on this basis alone.) Biopsies cannot be collected in
subjects with a single measurable lesion, even if accessible.
[0210] Target Population [0211] a) Subjects must have histologic or
cytologic confirmation of an incurable solid malignancy that is
advanced (metastatic and/or unresectable): [0212] i) Part A Dose
Escalation: BMS-986016 monotherapy [0213] (1) All solid tumor
histologies are permitted except for subjects with primary CNS
tumors [0214] (2) Only subjects without prior exposure to immune
cell-modulating antibody regimens (ICMARs) such as, but not limited
to, CTLA-4, ipilimumab, tremelimumab, anti-PD-1, anti-PD-L1,
anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40 antibodies, are
allowed; [0215] (3) Subjects must have received, and then
progressed or been intolerant to, at least one standard treatment
regimen in the advanced or metastatic setting, if such a therapy
exists. [0216] ii) Part B Dose Escalation: BMS-986016+BMS-936558
[0217] (1) All solid tumor histologies are permitted except for
subjects with primary CNS tumors. Subjects with or without prior
anti-PD-1 or anti-PD-L1 antibody therapy are eligible.
Alternatively, all solid tumor histologies naive to ICMARs such as,
but not limited to, anti-CTLA-4, anti-PD-1, anti-PD-L1, anti-PD-L2,
anti-KIR, anti-CD137, or anti-OX40 antibodies, will be permitted
except for subjects with primary CNS tumors. [0218] (2) Subjects
without prior anti-PD-1 or anti-PD-L1 antibody therapy cannot have
had prior exposure to any other ICMARs such as, but not limited to,
ipilimumab, tremelimumab, anti-PD-L2, anti-KIR, anti-CD137, or
anti-OX40 antibodies. Alternatively, Subjects without prior
anti-PD-1 or anti-PD-L1 antibody therapy cannot have had prior
exposure to any other ICMARs such as, but not limited to,
ipilimumab, tremelimumab, anti-PD-L2, anti-KIR, anti-CD137, or
anti-OX40 antibodies. [0219] (3) NSCLC subjects whose disease
progresses while-on or after therapy with anti-PD-1 or anti-PD-L1
antibody as most recent therapy. Alternatively, subjects with prior
anti-PD-1 or anti-PD-L1 antibody as most recent therapy: [0220] (a)
Disease is nonresponsive while on anti-PD-1 or anti-PD-L1 antibody
therapy and presents with PD (per RECIST) within 16 weeks of
starting therapy. [0221] (b) Cannot have had therapy discontinued
due to serious and/or life-threatening anti-PD-1 or anti-PD-L1
antibody-related toxicity (e.g., dose-limiting toxicity in prior
study). [0222] (c) Must provide informed consent within 60 days
after the last dose of anti-PD-1 or anti-PD-L1 antibody therapy.
[0223] (d) Cannot have had prior exposure to ICMARs such as, but
not limited to, anti-CTLA-4 antibody therapy, ipilimumab,
tremelimumab, anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40
antibodies. [0224] (4) Melanoma subjects whose disease is
progressing while-on or after receiving anti-CTLA-4 and anti-PD-1
or anti-PD-L1 antibody therapies (a) Anti-CTLA-4 and anti-PD-1 or
anti-PD-L1 antibody therapies could have been received in
sequential or combination regimens (b) Last dose of anti-CTLA-4
antibody therapy must have been received .gtoreq.100 days of first
dose of study medication (c) Cannot have had therapy discontinued
due to serious and/or life-threatening antibody-related toxicity
(e.g., dose-limiting toxicity in prior study) (d) Cannot have had
prior exposure to any ICMARs other than anti-CTLA-4 and anti-PD-1
or anti-PD-L1 antibody therapy. [0225] (5) Subjects must have
received, and then progressed or been intolerant to at least one
standard treatment regimen. [0226] iii) Part C Cohort Expansion
[0227] (1) The following groups are enrolled: [0228] (a)
Melanoma--subjects naive to ICMARs [0229] (b) Melanoma--subjects
whose disease is nonresponsive while on anti-PD-1 or anti-PD-L1
antibody therapy as most recent therapy and presents with PD (per
RECIST) within 16 weeks of starting therapy. Subject must provide
informed consent within 60 days after last dose of anti-PD-1 or
anti-PD-L1 antibody therapy and should have not discontinued
antibody therapy due to serious and/or life-threatening toxicity
(e.g., dose-limiting toxicity in prior study). These subjects
cannot have had prior exposure to any other ICMARs such as, but not
limited to, ipilimumab, tremelimumab, anti-PD-L2, anti-KIR,
anti-CD137, or anti-OX40 antibodies. Alternatively, subjects whose
disease is progressing while on or after receiving anti-CTLA-4 and
anti-PD-1 or anti-PD-L1 antibody therapy (in sequential or
combination regimens), are eligible. Last dose of anti-CTLA-4
antibody therapy must have been received .gtoreq.100 days of first
dose of study medication. Subjects should have not discontinued
antibody therapy due to serious and/or life-threatening toxicity
(e.g., dose-limiting toxicity in prior study). These subjects
cannot have had prior exposure to any ICMARs other than anti-CTLA-4
and PD-1 or anti-PD-L1 antibody therapy. [0230] (c) Non-small cell
lung cancer (NSCLC)--subjects naive to ICMARs [0231] (d)
NSCLC--subjects whose disease is nonresponsive while on anti-PD-1
or anti-PD-L1 antibody therapy as most recent therapy and presents
with PD (per RECIST) within 16 weeks of starting therapy. Subject
must provide informed consent within 60 days after last dose of
anti-PD-1 or anti-PD-L1 antibody therapy and should have not
discontinued antibody therapy due to serious and/or
life-threatening toxicity (e.g., dose-limiting toxicity in prior
study). These subjects cannot have had prior exposure to any other
ICMARs such as, but not limited to, ipilimumab, tremelimumab,
anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40 antibodies.
Alternatively, NSCLC subjects whose disease progresses while-on or
after therapy with anti-PD-1 or anti-PD-L1 antibody as most recent
therapy. Subject should have not discontinued antibody therapy due
to serious and/or life-threatening toxicity (e.g., dose-limiting
toxicity in prior study). These subjects cannot have had prior
exposure to any other ICMARs such as, but not limited to,
anti-CTLA-4, anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40
antibodies. [0232] (e) HPV-associated head and neck
tumors--subjects naive to ICMARs and with HPV positivity as defined
by p16 immunohistochemistry (IHC)-positive and/or HPV-16 in situ
hybridization (ISH)-positive (i) Histology restricted to squamous
cell carcinoma. Alternatively, subjects with advanced/metastatic
head and neck tumors naive to ICMARs (i) Histology restricted to
squamous cell carcinoma. [0233] (f) Gastric adenocarcinoma subjects
naive to ICMARs (i) HER2(+) and HER2(-) subjects are allowed [0234]
b) Subjects must have received, and then progressed or been
intolerant to, at least one standard treatment regimen in the
advanced or metastatic setting, if such a therapy exists. [0235] c)
Subjects with any prior treatment regimens are eligible. The
following are not considered separate lines of treatment: addition
of a compound to an ongoing regimen, restarting the same regimen
after a drug holiday, or switching from IV to oral therapy. [0236]
d) Presence of at least one lesion with measurable disease as
defined by RECIST v1.1 criteria for response assessment. Subjects
with lesions in a previously irradiated field as the sole site of
measurable disease are permitted to enroll provided that the
lesion(s) have demonstrated clear progression prior to the time of
informed consent and can be measured accurately. [0237] e) ECOG
status of 0 or 1. [0238] f) Life expectancy of .gtoreq.12 weeks at
the time of informed consent. [0239] g) Adequate organ function as
defined by the following: [0240] i) White blood cells (WBCs)
.gtoreq.2000/.mu.L (stable off any growth factor within 4 weeks of
first study drug administration) [0241] ii) Neutrophils
.gtoreq.1500/.mu.L (stable off any growth factor within 4 weeks of
first study drug administration) [0242] iii) Platelets
.gtoreq.100.times.10.sup.3/.mu.L (transfusion to achieve this level
is not permitted within 2 weeks of first study drug administration)
[0243] iv) Hemoglobin .gtoreq.8.5 g/dL (transfusion to achieve this
level is not permitted within 2 weeks of first study drug
administration) [0244] v) Creatinine <1.5.times.ULN or
creatinine clearance .gtoreq.40 mL/min (Cockcroft-Gault formula)
[0245] vi) ALT and AST.ltoreq.3.times.ULN [0246] vii) Lipase and
amylase <1.5.times.ULN [0247] viii) Total bilirubin
.ltoreq.1.5.times.ULN (except subjects with Gilbert's Syndrome who
must have normal direct bilirubin) [0248] h) Normal thyroid
function, or stable on hormone supplementation [0249] i) Ability to
comply with treatment, PK, and pharmacodynamic sample collection
and required study follow-up. [0250] j) Subject re-enrollment: This
study permits the re-enrollment of a subject that has discontinued
the study as a pre-treatment failure (i.e., subject has not been
randomized/has not been treated). If re-enrolled, the subject must
be re-consented.
[0251] Age and Reproductive Status [0252] a) Men and women, ages
.gtoreq.18 years at the time of informed consent [0253] b) Women of
childbearing potential (WOCBP) must use methods of contraception.
For a teratogenic study drug and/or when there is insufficient
information to assess teratogenicity (preclinical studies have not
been done), 2 forms of contraception are required. One method must
be highly effective (failure rate of less than 1% when used
consistently and correctly) and the second method may also be
highly effective. The individual methods of contraception should be
determined in consultation with the Investigator. WOCBP must follow
instructions for birth control for a total of 24 weeks after the
last dose of investigational drug (a period of 30 days plus the
time required for the investigational drug to undergo 5
half-lives). Women of childbearing potential (WOCBP) are defined as
any female who have experienced menarche and who has not undergone
surgical sterilization (hysterectomy or bilateral oophorectomy) and
is not postmenopausal. Menopause is defined as 12 months of
amenorrhea in a woman over age 45 in the absence of other
biological or physiological causes. In addition, females under the
age of 55 must have a documented serum follicle-stimulating hormone
(FSH) level >40 mIU/mL to confirm menopause. Females treated
with hormone replacement therapy, (HRT) are likely to have
artificially suppressed FSH levels and may require a washout period
in order to obtain a physiologic FSH level. The duration of the
washout period is a function of the type of HRT used. The duration
of the washout period below are suggested guidelines and the
investigators should use their judgement in checking serum FSH
levels. If the serum FSH level is >40 mlU/ml at any time during
the washout period, the woman can be considered postmenopausal.
[0254] 1 week minimum for vaginal hormonal products, (rings,
creams, gels) [0255] 4 week minimum for transdermal products [0256]
8 week minimum for oral products [0257] Other parenteral products
may require washout periods as long as 6 months. [0258] c) Women
must have a negative serum or urine pregnancy test (minimum
sensitivity of urine pregnancy test of 25 IU/L of either total
human chorionic gonadotropin (hCG) or the beta fraction) within 24
hours prior to the start of investigational product. [0259] d)
Women must not be breastfeeding. [0260] e) Men who are sexually
active with WOCBP must use methods of contraception. For a
teratogenic study drug and/or when there is insufficient
information to assess teratogenicity (preclinical studies have not
been done), 2 forms of contraception are required. One method must
be highly effective (failure rate of less than 1% when used
consistently and correctly) and the second method may also be
highly effective. Men who are sexually active with WOCBP must
follow instructions for birth control for a total of 33 weeks after
the last dose of investigational drug (a period of 90 days plus the
time required for the investigational drug to undergo 5
half-lives).
[0261] Women who are not of childbearing potential (i.e., who are
postmenopausal or surgically sterile; and permanently azoospermic
men (e.g., bilateral orchiectomy) do not require contraception.
Women of childbearing potential (WOCBP) are defined as females who
have experienced menarche and who have not undergone surgical
sterilization (hysterectomy or bilateraloophorectomy) or are not
postmenopausal. Menopause is defined clinically as 6 months of
amenorrhea in a woman over age 45 in the absence of other
biological or physiological causes. In addition, women under the
age of 55 must have a documented serum follicle-stimulating hormone
(FSH) level >40 mIU/mL to confirm menopause.
[0262] 7. Exclusion Criteria
[0263] Target Disease Exceptions
[0264] Subjects with known or suspected CNS metastases or with the
CNS as the only site of active disease are excluded with the
following exceptions: [0265] i) Subjects with controlled brain
metastases are allowed to enroll. Controlled brain metastases are
defined as those with no radiographic progression for at least 4
weeks after radiation and/or surgical treatment at the time of
consent. Subjects must have been off of steroids for at least 2
weeks prior to informed consent, and have no new or progressive
neurological signs and symptoms. [0266] ii) Subjects with signs or
symptoms of brain metastases are not eligible unless brain
metastases are ruled out by computed tomography (CT) or magnetic
resonance imaging (MRI).
[0267] Participation in any prior clinical study with BMS-936558,
including subjects in comparator arms, in which overall survival is
listed as the primary or co-primary endpoint and which has not
completed analysis based on the primary endpoint.
[0268] Medical History and Concurrent Diseases
[0269] Subjects with a prior malignancy are excluded, except
adequately treated basal cell or squamous cell skin cancer,
localized prostate cancer, carcinoma in situ of the cervix or
carcinoma in situ of the bladder, or in situ ductal or lobular
carcinoma of the breast. Subjects with other prior malignancies
diagnosed more than 2 years previously (at the time of informed
consent) who have received therapy with curative intent with no
evidence of disease during the interval who are considered to
present a low risk for recurrence are eligible.
[0270] Subjects with any active autoimmune disease or history of
known or suspected autoimmune disease with the exception of
subjects with isolated vitiligo, resolved childhood asthma/atopy,
controlled hypoadrenalism or hypopituitarism, and euthyroid
patients with a history of Grave's disease (subjects with
controlled hyperthyroidism must be negative for thyroglobulin and
thyroid peroxidase antibodies and thyroid-stimulating
immunoglobulin prior to study drug administration).
[0271] A known or underlying medical condition that could make the
administration of study drug hazardous to the subject or could
adversely affect the ability of the subject to comply with or
tolerate study.
[0272] Requirement for daily supplemental oxygen.
[0273] Uncontrolled or significant cardiovascular disease
including, but not limited to, any of the following: Myocardial
infarction or stroke/transient ischemic attack (TIA) within the 6
months prior to consent, uncontrolled angina within the 3 months
prior to consent, any history of clinically significant arrhythmias
(such as ventricular tachycardia, ventricular fibrillation, or
torsades de pointes), QTc prolongation >480 msec, history of
other clinically significant heart disease (i.e., cardiomyopathy,
congestive heart failure with New York Heart Association [NYHA]
functional classification pericarditis, significant pericardial
effusion).
[0274] Cardiovascular disease-related requirement for daily
supplemental oxygen.
[0275] A confirmed history of encephalitis, meningitis, or
uncontrolled seizures in the year prior to informed consent.
[0276] Positive blood screen for human immunodeficiency virus (HIV)
or known acquired immunodeficiency syndrome (AIDS).
[0277] History of any chronic hepatitis as evidenced by positive
test for hepatitis A antibody (HepA IgM) (Note: history of resolved
hepatitis A virus infection is not an exclusion criterion),
positive test for hepatitis B surface antigen (HBsAg) and/or
hepatitis B core antigen, positive test for qualitative hepatitis C
viral load (by PCR).
[0278] Evidence of active infection that requires systemic
antibacterial, antiviral, or antifungal therapy .ltoreq.7 days
prior to initiation of study drug therapy.
[0279] Any other significant acute or chronic medical illness.
[0280] Subjects who are unable to undergo venipuncture and/or
tolerate venous acces.
[0281] Any other sound medical, psychiatric, and/or social
reason.
[0282] Any of the following procedures or medications:
[0283] Within 2 weeks prior to time of informed consent: systemic
or topical cortico steroids at immunosuppressive doses 7.5 mg/day
of prednisone or equivalent), palliative radiation and gamma knife
radiosurgery in CNS, or medicinal herbal preparations.
[0284] Within 4 weeks prior to study drug administration: any
investigational drug or placebo, any anticancer therapy
(chemotherapy, biologics, therapeutic vaccines, radiotherapy, or
hormonal treatment), non-oncology vaccines containing live virus,
allergen hyposensitization therapy, growth factors, e.g.,
granulocyte-colony stimulating factor (G-CSF), granulocyte
macrophage-colony stimulating factor (GM-CSF), erythropoietin,
major surgery, or biphosphonates.
[0285] Within 10 weeks prior to study drug administration: receptor
activator of nuclear factor kappa-B ligand (RANK-L) inhibitors.
[0286] Allergies and Adverse Drug Reaction
[0287] History of allergy to anti-PD-1 or anti-PD-L1 antibody
therapy or to other monoclonal antibodies or related compounds or
to any of their components (e.g., history of severe
hypersensitivity reactions to drugs formulated with polysorbate
80).
[0288] Other Exclusion Criteria
[0289] Prisoners or subjects who are involuntarily
incarcerated.
[0290] Subjects who are compulsorily detained for treatment of
either a psychiatric or physical (e.g., infectious disease)
illness.
[0291] Inability to comply with restrictions and prohibited
activities and treatments.
[0292] 8. Guidelines for Dose Modification
[0293] Intrasubject dose escalation of BMS-986016 or BMS-936558 is
not permitted in this study. With the possible exception of
subjects being treated at a dose level that is subsequently deemed
to exceed the MTD, intrasubject dose reduction of BMS-986016 or
BMS-936558 is not permitted.
[0294] In some cases, the natural history of select adverse events
associated with immunotherapy can differ from and be more severe
than adverse events caused by other therapeutic classes. Early
recognition and management mitigates severe toxicity.
[0295] Additionally, management algorithms can assist with select
toxicities. Toxicities for which management algorithms have been
developed include:
[0296] Pulmonary
[0297] Gastrointestinal
[0298] Hepatic
[0299] Endocrine
[0300] Renal
[0301] Dermatologic
[0302] Neurologic
[0303] Subjects who experience the following must have all study
drug(s) held: [0304] DLTs (per definition, are related to study
drug) [0305] Select drug-related adverse events and drug-related
laboratory abnormalities: [0306] .gtoreq.Grade 1 pneumonitis [0307]
.gtoreq.Grade 2 abnormality in AST, ALT, total bilirubin, amylase,
or lipase [0308] .gtoreq.Grade 2 creatinine [0309] .gtoreq.Grade 2
diarrhea or colitis [0310] .gtoreq.Grade 2 neurological adverse
event [0311] Adverse event, laboratory abnormality, or concurrent
illness that, in the judgment of the Investigator, warrants
delaying the dose of study drug.
[0312] Dose delays >7 days are considered missed and is not be
replaced.
[0313] 9. Safety Assessments
[0314] Adverse events are assessed continuously during the study
and for 135 days after the last treatment. Adverse events are
evaluated according to the NCI CTCAE version 4.0. Adverse events
are coded using the most current version of Medical Dictionary for
Regulatory Activities (MedDRA) and reviewed for potential
significance and importance.
[0315] 10. Other Analyses
[0316] Various serologic tumor markers, gene mutation status, and
additional analyses are required dependent upon the subject's tumor
type as listed below in Table 4. With the exception of the
serologic tumor markers, the assessments are not performed if the
lab results from previous testing are available.
TABLE-US-00007 TABLE 4 Biomarkers by Tumor Type Tumor Type Study
Part Matrix Lab Test Assessment Timepoint Colorectal Parts A, B
Blood Serologic Tumor CEA.sup.a Multiple ONLY Marker Parts A, B
Tumor Gene Mutation EGFR.sup.b Screening ONLY Tissue Status K-RAS
MSI.sup.c Gastric Parts A, B, C Blood Serologic Tumor CEA.sup.a
Multiple Marker Parts A, B, C Tumor Gene Mutation HER-2.sup.d
Screening Tissue Status Parts A, B, C Tumor Real Time EBV.sup.f
Screening Tissue qPCR.sup.e Germ Cell Parts A, B Blood Serologic
Tumor .beta.hCG.sup.g Multiple ONLY Marker AFP.sup.h Head and Part
C ONLY Tumor IHC and/or ISH.sup.i HPV.sup.j Screening Neck Tissue
(Eligibility) Melanoma Parts A, B, C Tumor Gene Mutation BRAF
Screening Tissue Status NSCLC Parts A, B, C Tumor Gene Mutation
ALK.sup.k Screening Tissue Status K-RAS EGFR.sup.b Ovarian Parts A,
B Blood Serologic Tumor CA125.sup.l Multiple ONLY Marker Prostate
Parts A, B Blood Serologic Tumor PSA.sup.m Multiple ONLY Marker
.sup.aCEA: carcinoembryonic antigen .sup.bEGFR: epidermal growth
factor receptor .sup.cMSI: microsatellite instability .sup.dHER-2:
human epidermal growth factor receptor 2 status via IHC and/or ISH
.sup.eReal time qPCR: real time quantitative polymerase chain
reaction for BamH1-A Reading Frame-1(BARF1) gene .sup.fEBV:
Epstein-Barr virus .sup.g.beta.hCG: beta-human chorionic
gonadotrophin .sup.hAFP: alpha-fetoprotein .sup.iIHC and/or ISH:
p16 immunohistochemistry (IHC) and/or HPV-16 in situ hybridization
(ISH) .sup.jHPV: human papilloma virus .sup.kALK: anaplastic
lymphoma kinase .sup.lCA125: cancer antigen 125 .sup.mPSA: prostate
specific antigen
[0317] Additional measures, including non-study-required laboratory
tests, are performed as clinically indicated. Results of all
laboratory tests required by this protocol are recorded.
[0318] 11. Efficacy Assessments
[0319] Efficacy is evaluated in Parts A and B (dose escalation), as
well as in Part C (cohort expansion). Changes in tumor measurements
and tumor response at the time of each assessment are determined.
The baseline assessment during the screening period requires CT or
MRI scans of the chest, abdomen, and pelvis, and other anatomic
regions as indicated by individual subject's tumor type and/or
disease history. Subsequent timepoints require scans of the chest,
abdomen, and pelvis, as well as other anatomic regions that were
scanned at baseline based on the individual subject's tumor type
and/or disease history. Scans of the brain are otherwise required
as clinically indicated.
[0320] Analysis of response endpoints are performed according to
immune-related response criteria, irRECIST that reflect the
clinical experience with other T cell-directed immunotherapies in
which objective and durable responses were observed in subjects
following progression and without intervening alternative
anticancer therapy (Wolchok J D, et al., Clin. Can. Res. 2009;
15(23):7412-7420). Individual subject's best overall response
(BOR), duration of progression-free survival (PFS), and duration of
response (DOR) is calculated as appropriate.
[0321] Tumor status are assessed at baseline, during treatment
(every 8 weeks) for up to twelve 8-week cycles of therapy, and once
during follow-up. CT and MRI scans re read and assessed locally per
RECIST v1.1. All imaging scans are de-identified and archived in
their native Digital Imaging and Communications in Medicine (DICOM)
format as part of the subject's study file.
[0322] The efficacy assessments include the ORR (e.g., PR+CR), DOR,
and PFSR at landmark timepoints (e.g., 24 weeks), based on
assessment of tumor response using irRECIST and RECIST v1.1.
Landmark 2-year overall survival (OS).
[0323] 12. Pharmacokinetic Assessments
[0324] Serum samples for BMS-986016 pharmacokinetics and anti-drug
antibody (ADA) assessments are collected for all subjects. Serum
samples for BMS-936558 pharmacokinetics and ADA assessments re
collected for all subjects enrolled in Part B and C. The serum
samples are analyzed for BMS-986016 and BMS-936558 by a validated
immunoassay. In addition, selected serum samples are analyzed by an
exploratory orthogonal method (e.g., liquid chromatography
[LC]-mass spectrometry [MS]/MS) that measures total BMS-986016
and/or BMS-936558.
[0325] 13. Exploratory Biomarker Assessment
[0326] The pharmacodynamics of BMS-986016 treatment administered
alone or in combination with BMS-936558 is assessed by quantifying
biomarkers in peripheral blood and tumor tissue in the first 3
subjects enrolled at each dose level during the dose escalation
(Parts A and B) and in subjects with melanoma and head and neck
cancers during cohort expansion (Part C) phases of the study.
Detailed schedules of pharmacodynamic evaluations are provided
below in Tables 5-6. Details regarding the tumor tissues
requirements for subjects in Parts A, B, and C of the study are
provided below in Table 7.
TABLE-US-00008 TABLE 5A Part A & B (Dose Escalation) -
Biomarker Sampling Schedule (ONLY for First 3 Subjects in each Dose
Level) PBMC Collection Serum Immunophenotyping/ Ex vivo Tumor Whole
Blood Timing Soluble Biomarkers Tetramer Functional Assay Archival
Gene Expression Study Day (Serum Biomarkers) (Flow Cytometry/PBMC)
(Cellular Assay) Tissue.sup.a (Whole Blood mRNA) SNP Screening X
Cycle 1 Day 1 X X X X X Day 5.sup.a X Day 8 X X Day 15 X X X Day 29
X X X Day 43 X X X X Cycle 2 Day 29 X X X X Upon Progression Upon X
X X X X Progression.sup.b .sup.aDay 5 visit can occur on Day 3 or
Day 4 .sup.bOptional; collected upon confirmation of PD NOTE: All
samples are drawn pre-dose
TABLE-US-00009 TABLE 5B Part A & B (Dose Escalation) -
Biomarker Sampling Schedule (ONLY for First 3 Subjects in each Dose
Level) PBMC Collection Serum Immunophenotyping/ Ex vivo Tumor Whole
Blood Timing Soluble Biomarkers Tetramer Functional Assay Archival
Gene Expression Study Day (Serum Biomarkers) (Flow Cytometry/PBMC)
(Cellular Assay) Tissue (Whole Blood mRNA) SNP Screening X Cycle 1
Day 1 X X X X X Day 5.sup.b X Day 8 X X Day 15 X X X Day 29 X X X
Day 43 X X X X Cycle 2 Day 29 X X X X Upon Progression Upon X X X X
X Progression.sup.b Upon Drug-related AE Upon X X X occurrence of
.gtoreq.Grade 2 drug related pneumonitis or neurological AE a Day 5
visit can occur on Day 3 or Day 4. Day 8 visit can occur on Day 7
or Day 9. .sup.bOptional; collected upon confirmation of PD NOTE:
All samples are drawn pre-dose
TABLE-US-00010 TABLE 6A Part C (Cohort Expansion) - Biomarker
Sampling Schedule (ONLY for Subjects with Melanoma and Head and
Neck Cancer) PBMC Collection Serum Immunophenotyping/ Ex vivo Tumor
Whole Blood Timing Soluble Biomarkers Tetramer Functional Assay
"Fresh" Gene Expression Study Day (Serum Biomarkers) (Flow
Cytometry/PBMC) (Cellular Assay) Tumor Biopsy (Whole Blood mRNA)
SNP Screening X.sup.a Cycle 1 Day 1 .sup. X.sup.b X X X X Day
5.sup.c X Day 8 X X Day 15 X X X Day 29 X X X Day 43 X X X X Day
50-56 X.sup.d Cycle 2 Day 29 X X X X Upon Progression Upon X X X
X.sup. X Progression.sup.e NOTE: All samples are drawn pre-dose
.sup.aFresh tumor biopsy is mandatory for subjects with melanoma
and head and neck cancer in Part C. .sup.bSerum and plasma at Cycle
1 Day 1. Serum only at all other timepoints. .sup.cDay 5 visit can
occur on Day 3 or Day 4 .sup.dFresh tumor biopsy is mandatory for
subjects with melanoma and head and neck cancer in Part C. Biopsy
is obtained at anytime during Cycle 1, Week 8 (Days 50-56) at the
same time as diagnostic imaging. .sup.eOptional; collected upon
confirmation of PD
TABLE-US-00011 TABLE 6B Part C (Cohort Expansion) - Biomarker
Sampling Schedule (ONLY for Subjects with Melanoma and Head and
Neck Cancer) PBMC.sup.b Collection Serum Plasma Immunophenotyping/
Ex vivo Tumor Whole Blood Timing Soluble (Serum Tetramer (Flow
Functional Assay "Fresh" Gene Expression Study Day Biomarkers
Biomarkers) Cytometry/PBMC) (Cellular Assay) Tumor Biopsy (Whole
Blood mRNA) SNP Screening X.sup.a Cycle 1 Day 1 X X X X X X Day
5.sup.c X Day 8.sup.c X X Day 15 X X X Day 29 X X X X Day 36 X X X
Day 43 X X X X X Day 50-56 X.sup.d Cycle 2 Day 29 X X X X Upon
Progression Upon X X X X.sup. X Progression.sup.c Upon Drug-related
AE Upon X X X occurrence of .gtoreq.Grade 2 drug related
pneumonitis or neurological AE .sup.aFresh tumor biopsy is
mandatory for subjects with melanoma and head and neck cancer Part
C. .sup.bPBMC samples only to be collected for subjects in the US,
not required for subjects Ex-US. .sup.c Day 5 visit can occur on
Day 3 or Day 4. Day 8 visit can occur on Day 7 or Day 9.
.sup.dFresh tumor biopsy is mandatory for subjects with melanoma
and head and neck cancer in Part C. Biopsy can be obtained at
anytime during Cycle 1, Week 8 (Days 50-56) at same time as
diagnostic imaging. e Optional; to be collected upon confirmation
of PD.
TABLE-US-00012 TABLE 7 Tumor Tissue Requirements for Parts A, B,
and C Part A and B Part C Study Part (Dose Escalation) (Cohort
Expansion) Subjects ALL subjects in Part A Subjects with Subjects
with or B melanoma or head and NSCLC or gastric neck tumors
adenocarcinoma ONLY ONLY Type of Archived tumor tissue. Mandatory
"fresh" Archived tumor Specimen If archived sample is not biopsies
(pre- and on- tissue. available, must obtain a treatment) If
archived "fresh" pre-treatment sample is not tumor biopsy
available, must obtain a "fresh" pre-treatment tumor biopsy Upon
Optional "fresh" biopsy Optional "fresh" biopsy Optional "fresh"
Progression upon confirmation of upon confirmation of biopsy upon
PD PD confirmation of PD
[0327] Soluble Biomarkers (Serum Biomarkers)--Parts A, B, and C
[0328] Pre-treatment and on-treatment serum levels of chemokines,
cytokines, and tumor-associated soluble proteins is assessed by
techniques that include, but are not limited to, ELISA or multiplex
assays. Analytes include markers of inflammation, immune
activation, host tumor growth factors, and tumor-derived
proteins.
[0329] Antitumor Antibodies (Serum Biomarkers)--Parts A, B, and
C
[0330] Treatment with BMS-986016 and BMS-936558 may result in the
generation of novel, or an increase in existing, antibodies to
tumor-associated antigens. An assessment of antibodies to a panel
of >8000 proteins is performed using pre-treatment and
on-treatment serum in multiplex and ELISAs. These data are used to
explore if antitumor antibodies are associated with clinical
response and safety parameters, as well as inform pharmacodynamics
of drug administration.
[0331] Immunophenotyping (Flow Cytometry/PBMC)--Parts A, B, and
C
[0332] Peripheral blood mononuclear cells (PBMCs) are used to
characterize and quantify the activation and regulatory status of
myeloid and lymphoid cells by polychromatic flow cytometry. Subsets
of cells characterized by immunophenotyping include naive,
activated, and exhausted effector and memory T cell populations,
regulatory T cells, and myeloid-derived suppressor cells.
[0333] Ex Vivo Functional Assays (Cellular Assay)--Parts A, B, and
C
[0334] To assess whether BMS-986016 and BMS-936558 restores T cell
activation and function, PBMCs are isolated and cryopreserved. The
functional status of effector T cells, including, but not limited
to, IFN-.gamma. and granzyme B, is assessed by flow cytometric
staining.
[0335] Peripheral Blood Gene Expression (Whole Blood mRNA) and
Tumor Gene Expression--Parts A, B, and C
[0336] The expression level of genes related to response to
BMS-986016.+-.BMS-936558 is quantified by microarray and/or
quantitative reverse transcription polymerase chain reaction
(RT-PCR) analysis in whole blood and tumor samples. Analysis
includes, but is not necessarily be limited to, genes encoding
BMS-986016-stimulated effector functions (perforin, granzyme B, and
IFN-.gamma.) and genes encoding T cell co-stimulatory receptors
(PD-1, PD-L1, and CTLA-4).
[0337] Circulating Tumor DNA Analysis (Serum (Plasma)
Biomarkers)--Part C
[0338] The presence of cell-free DNA in circulating blood is a
well-documented phenomenon. Fragments of DNA are shed into the
blood stream from dividing cells during cell proliferation or cell
death. In patients with cancer, a fraction of this DNA is
tumor-derived and is termed circulating tumor DNA (ctDNA). Albeit
small, fragments of DNA average between 180 to 200 bp and specific
genomic regions can be amplified with PCR. Moreover, several
studies have detected mutations in ctDNA that exactly correspond to
mutations from the parent tumor. Using tissue and plasma from
patients with known driver mutations in melanoma or head and neck
cancer, BEAMing technology is utilized to count the frequency of
mutations in circulation.
[0339] Single Nucleotide Polymorphism Analysis (SNP)--Parts A, B,
and C
[0340] In order to identify potential polymorphisms associated with
the safety and efficacy of BMS-986016 selected genes are evaluated
for single nucleotide polymorphisms (SNPs). Genes of interest
include, but are not limited to, PD-1, PD-L1, MHC class II, LAG-3,
and CTLA-4.
[0341] Tumor Biopsy Analysis--Parts A and B
[0342] Tumor tissue is collected from all subjects in the dose
escalation portion of the protocol. Immunohistochemistry is used to
assess the number and composition of immune infiltrates to define
the immune cell subsets present within FFPE tumor tissue before and
potentially after exposure to BMS-986016 and BMS-936558. These IHC
analyses include, but are not necessarily be limited to, the
following markers: CD4, CD8, LAG-3, MHC II, PD-1, PD-L1, and PD-L2.
Correlations between gene expression and IHC expression are made
between assays performed if deemed to be informative.
[0343] Tumor-Based Biomarker Measures--Part C
[0344] Paired pre- and on-treatment tumor biopsies are mandatory
for all subjects with melanoma or head and neck cancer who are
enrolled in Part C (cohort expansion). Subjects for whom adequate
paired pre- and on-treatment biopsies are not collected may be
replaced.
[0345] Subjects have at least one lesion large enough to undergo
repeated biopsies (pre- and on-treatment biopsies) via core needle
(minimum size 18 gauge) or have at least 2 distinct lesions
eligible for core needle or excisional biopsies. The expected core
needle length should be greater than 5 mm. A punch biopsy is
acceptable for cutaneous lesions. Fine needle aspirate biopsies are
not accepted. At least two core biopsies are taken at each
timepoint, but collection of additional cores is strongly
encouraged if deemed clinically safe by the investigator. An
assessment of biopsy quality by a pathologist is strongly
encouraged at the time of the procedure. All biopsies collected
must have a detailed pathology report submitted with the
specimen.
[0346] Tumor biopsy specimens are obtained from consenting subjects
prior to and during treatment with BMS-986016 and BMS-936558 to
characterize immune cell populations and expression of selected
tumor markers. Biopsy samples are used for the following
assessments: [0347] Characterization of TILs and tumor antigens.
Immunohistochemistry is used to assess the number and composition
of immune infiltrates to define the immune cell subsets present
within FFPE tumor tissue before and after exposure to BMS-986016
and nivolumab. These IHC analyses include, but are not necessarily
limited to, the following markers: CD4, CD8, LAG-3, MHC II, PD-1,
PD-L1, and PD-L2. Correlations between gene expression and IHC
expression are made between assays performed if deemed to be
informative. [0348] Laser capture microdissection. Isolation of
tumor and/or TIL on FFPE sections is performed by laser capture
microdissection (LCM) for high-throughput profiling of molecular
events within the tumor microenvironment. [0349] Characterization
of T cell repertoire. DNA sequencing is performed on pre- and
post-treatment FFPE tumor tissue to assess the composition of the T
cell repertoire. Low T cell receptor diversity may be a poor
prognostic factor of overall survival in metastatic breast cancer
patients. Currently, there is a poor understanding of T cell
receptor diversity as a predictor factor of response to
immunotherapy, given that the major mechanism of BMS-936558 and
BMS-986016 is hypothesized to be the functional restoration of T
cell antitumor immunity. Therefore, a characterization of the
diversity of the T cell compartment in the periphery, and within
the tumor, at baseline and on-treatment is performed by T cell
receptor next-generation DNA sequencing. T cell repertoire analysis
is also performed from DNA isolated from peripheral blood to
compare the status of tumor and peripheral T cell repertoire pre
and post treatment. [0350] Gene expression profiling. Tumor
biopsies that are collected in RNAlater or similar reagent are
examined for mRNA gene expression by Affymetrix gene array
technology and/or quantitative real-time polymerase chain reaction
(qPCR) to detect expression of selected immune-related genes.
[0351] In situ cytokine and negative regulator expression. Tumors
biopsies are quantitatively evaluated for RNA, including CD3,
IFN-.gamma., LAG-3, and PD-1.
[0352] Subjects whose screening biopsy yields inadequate tissue
quantity or quality are allowed to continue in the study. If
on-treatment biopsy is not successful, subjects also continue on
study. Such subjects are replaced in order to obtain 48 subjects
with adequate paired tumor biopsies. If subjects have a response to
treatment, on-treatment biopsies might not be possible. In this
case, subjects also continue on study.
[0353] The tumor tissue that is obtained from these biopsies is
divided equally into FFPE and frozen samples, which can be used for
histologic confirmation of melanoma, as well as for the assays
listed above.
[0354] Biopsies are done with local anesthesia or conscious
sedation. Institutional guidelines for the safe performance of
biopsies are followed. Excisional biopsies are performed to obtain
tumor biopsy samples. Invasive procedures that require general
anesthesia are not performed to obtain a biopsy specimen. However,
if a surgical procedure is performed for a clinical indication,
excess tumor tissue is used for research purposes with the consent
of the subject.
[0355] 14. Immunogenicity Assessments
[0356] Serum samples collected at timepoints are analyzed by a
validated immunogenicity assay. Selected serum samples are analyzed
by an exploratory orthogonal method that measures anti-BMS-986016
or anti-BMS-936558. Potential results generated from any orthogonal
method are intended as informational for technology exploration
purposes and re not reported.
[0357] In addition, ad hoc serum samples designated for
pharmacokinetic or biomarker assessments are used for
immunogenicity analysis if required (e.g., insufficient volume for
complete immunogenicity assessment or to follow up on suspected
immunogenicity related adverse event).
[0358] 15. Adverse Events
[0359] An adverse event (AE) is defined as any new untoward medical
occurrence or worsening of a preexisting medical condition in a
clinical investigation subject administered an investigational
(medicinal) product and that does not necessarily have a causal
relationship with this treatment. An AE is therefore any
unfavorable and unintended sign (such as an abnormal laboratory
finding), symptom, or disease temporally associated with the use of
investigational product, whether or not considered related to the
investigational product.
[0360] The causal relationship to study drug is determined by a
physician and used to assess all adverse events (AE). The casual
relationship can be one of the following: [0361] Related: There is
a reasonable causal relationship between study drug administration
and the AE. [0362] Not related: There is not a reasonable causal
relationship between study drug administration and the AE.
[0363] The term "reasonable causal relationship" means there is
evidence to suggest a causal relationship.
[0364] Serious Adverse Events
[0365] A serious adverse event (SAE) is any untoward medical
occurrence that at any dose: [0366] results in death [0367] is
life-threatening (defined as an event in which the subject was at
risk of death at the time of the event; it does not refer to an
event which hypothetically might have caused death if it were more
severe) [0368] requires inpatient hospitalization or causes
prolongation of existing hospitalization [0369] results in
persistent or significant disability/incapacity [0370] is a
congenital anomaly/birth defect [0371] is an important medical
event (defined as a medical event(s) that may not be immediately
life-threatening or result in death or hospitalization but, based
upon appropriate medical and scientific judgment, may jeopardize
the subject or may require intervention (e.g., medical, surgical)
to prevent one of the other serious outcomes listed in the
definition above). Examples of such events include, but are not
limited to, intensive treatment in an emergency room or at home for
allergic bronchospasm; blood dyscrasias or convulsions that do not
result in hospitalization.) Potential drug induced liver injury
(DILI) is also considered an important medical event.
[0372] Suspected transmission of an infectious agent (e.g.,
pathogenic or nonpathogenic) via the study drug is an SAE. Although
pregnancy, overdose, cancer, and potential drug induced liver
injury (DILI) are not always serious by regulatory definition,
these events must be handled as SAEs. Any component of a study
endpoint that is considered related to study therapy (e.g., death
is an endpoint, if death occurred due to anaphylaxis, anaphylaxis
must be reported) is reported as SAE.
[0373] The following hospitalizations are not considered SAEs:
[0374] a visit to the emergency room or other hospital department
<24 hours, that does not result in admission (unless considered
an important medical or life-threatening event) [0375] elective
surgery, planned prior to signing consent [0376] admissions as per
protocol for a planned medical/surgical procedure [0377] routine
health assessment requiring admission for baseline/trending of
health status (e.g., routine colonoscopy) [0378] medical/surgical
admission other than to remedy ill health and planned prior to
entry into the study. Appropriate documentation is required in
these cases [0379] admission encountered for another life
circumstance that carries no bearing on health status and requires
no medical/surgical intervention (e.g., lack of housing, economic
inadequacy, caregiver respite, family circumstances, administrative
reason).
[0380] Following the subject's written consent to participate in
the study, all SAEs, whether related or not related to study drug,
are collected, including those thought to be associated with
protocol-specified procedures. All SAEs are collected that occur
during the screening period and within 135 days of discontinuation
of dosing. If applicable, SAEs are collected that relate to any
later protocol-specified procedure (e.g., a follow-up skin biopsy).
All SAEs are followed to resolution or stabilization.
[0381] Nonserious Adverse Events
[0382] A nonserious adverse event is an AE not classified as
serious. The collection of nonserious AE information begins at
initiation of study drug and continues for 135 days after
discontinuation of dosing. Nonserious AEs are followed to
resolution or stabilization, or reported as SAEs if they become
serious. Follow-up is also required for nonserious AEs that cause
interruption or discontinuation of study drug and for those present
at the end of study treatment as appropriate. All identified
nonserious AEs are recorded and described on the nonserious AE page
of the CRF (paper or electronic).
[0383] Completion of supplemental CRFs are requested for AEs and/or
laboratory abnormalities that are reported/identified during the
course of the study.
[0384] 16. Statistical Considerations
[0385] Sample Size Determination
[0386] Dose Escalation (Parts A and B): Sample size at each dose
depends on observed toxicity and cannot be precisely determined.
Part A and Part B have 3 to 9 subjects in each cohort.
[0387] Cohort Expansion (Part C): Cohort expansion allows for
better estimation of the toxicity rate and provide better precision
around preliminary estimates of efficacy. If .ltoreq.5 of 16
subjects (i.e., .about.30% of in a cohort) experience a toxicity,
there is at least 90% confidence that the true toxicity rate is not
greater than 50.4% (based on Clopper-Pearson exact binomial 1-sided
90% confidence interval). A sample size of 16 subjects per cohort
also allows for estimation of the proportion of subjects with
objective response (i.e., CR+PR) within a cohort such that the
maximum distance between the estimated rate and either limit of the
exact 2-sided 95% Clopper-Pearson confidence interval is 27.4%.
[0388] Populations for Analyses [0389] All Enrolled Subjects
Analysis Set: This analysis set contains all subjects (including
screen failures) who signed an informed consent for the study.
[0390] All Treated Subjects-Analysis set: This analysis set
includes all subjects who receive either drug. [0391]
Response-Evaluable Subjects: This analysis set includes all
subjects who receive either study drug, have a baseline tumor
assessment with measurable disease, and one of the following: (1)
at least one evaluable on-treatment tumor assessment, (2) clinical
progression, or (3) death prior to the first on-treatment tumor
evaluation. [0392] BMS-986016 Pharmacokinetic Analysis Set: This
analysis set includes all subjects who receive BMS-986016 and have
at least one valid PK parameter to be included in statistical
analyses of BMS-986016 PK data. [0393] BMS-986016 Immunogenicity
Analysis Set: This analysis set includes all subjects who receive
BMS-986016 and have at least one BMS-986016 immunogenicity sample
available. [0394] BMS-936558 Immunogenicity Analysis Set: This
analysis set includes all subjects who receive BMS-936558 and have
at least one BMS-936558 immunogenicity sample available. [0395]
Pharmacodynamic Analysis Set: This analysis set includes all
treated subjects for whom pharmacodynamic measurements are
available at baseline and at least one other timepoint.
[0396] Endpoints
[0397] The primary endpoint of this Phase 1 study is safety as
measured by the rate of AEs, serious adverse events (SAEs), deaths,
and laboratory abnormalities (e.g., Grade 3 or higher per CTCAE v
4), assessed during treatment and for up to 135 days of follow-up.
All subjects who receive at least one dose of BMS-986016 or
BMS-936558 are analyzed for safety.
[0398] The PK of BMS-986016 administered both alone and in
combination with BMS-936558 is assessed as a secondary objective
using the following endpoints derived from serum concentration
versus time data in Cycle 1 and Cycle 3: [0399] Cmax Maximum
observed serum concentration [0400] Tmax Time of maximum observed
serum concentration [0401] Ctrough Trough observed serum
concentration [0402] Ctau Concentration at the end of a dosing
interval (e.g., concentration at 336 hours) [0403] Css,avg Average
concentration over a dosing interval ([AUC(TAU)/tau] [0404]
AUC(TAU) Area under the concentration-time curve in one dosing
interval [0405] CLT Total body clearance [0406] Vss Volume of
distribution at steady state [0407] T-HALFeff AUC Effective
elimination half-life that explains the degree of AUC accumulation
observed [0408] T-HALFeff Cmax Effective elimination half-life that
explains the degree of Cmax accumulation observed [0409] AI_AUC
Accumulation index; ration of AUC(TAU) at steady state to AUC(TAU)
after the first dose [0410] AI_CMAS Cmax accumulation index; ratio
of Cmax at steady state to Cmax after the first dose [0411] AI_Ctau
Ctau accumulation index; ratio of Ctau at steady state to Ctau
after the first dose [0412] DF Degree of fluctuation or fluctuation
index ([Cmax -Ctau]/Css,avg)
[0413] Individual subject PK parameter values are derived by
noncompartmental methods by a validated PK analysis program. Actual
times are used for the analyses.
[0414] Efficacy
[0415] Efficacy is assessed as a secondary objective using the
endpoints described below for irRECIST and RECIST v1.1. For the
purposes of patient management, clinical decision making is based
on RECIST. Statistical analysis and reporting are based on both
criteria. [0416] Best overall response (BOR) is the best response
designation recorded from the start of the study treatment until
the last protocol specified tumor assessment (e.g., 30 day
follow-up visit) taking into account any requirement for
confirmation, based on RECIST v1.1 or irRECIST criteria. CR or PR
determinations included in the BOR assessment are confirmed by a
consecutive second (confirmatory) evaluation meeting the criteria
for response and performed at least 4 weeks after the criteria for
response are first met. [0417] Objective response rate (ORR) is
defined as the total number of subjects whose BOR is either CR or
PR divided by the total number of subjects in the population of
interest. [0418] Duration of response (DOR) computed only for
subjects with a BOR of CR or PR is defined as the number of days
between the date of first response and the subsequent date of
objectively documented disease progression based on the criteria
(RECIST v1.1 or irRECIST) or death, whichever occurs first. For
those subjects who remain alive and have not progressed or received
subsequent therapy, duration of response is censored on the date of
last protocol specified tumor assessment. Subjects who receive
subsequent therapy are censored at the start of subsequent therapy.
[0419] Progression free survival (PFS) is defined as the
probability of a subject remaining progression-free and surviving.
The probability is computed based on the number of days between the
first dose of study drug and progressive disease (as defined by
RECIST or irRECIST) or death. For those subjects who remain alive
and have not progressed, PFS is censored on the date of the last
protocol specified tumor assessment.
[0420] These endpoints are determined based on tumor measurements
occurring every 8 weeks during the Treatment Period (up to twelve
8-week cycles), and once during the Clinical Follow-up period (30
days), for a total of .about.1.9 years.
[0421] Immunogenicity
[0422] At the sample level, individual samples are characterized as
ADA-positive or ADA-negative. A subject is considered to have a
positive sample at baseline if the last sample prior to the
initiation of treatment is ADA-positive. For example, a
post-baseline sample from a subject who is ADA-negative at baseline
is considered ADA-positive if ADA is detected. A post-baseline
sample from a subject who is ADA-positive at baseline is considered
ADA-positive if there is a relevant increase in titer (magnitude of
the increase in titer considered relevant may vary by drug and
assay, and is delineated in the statistical analysis plan). At the
subject level, relevant ADA endpoints may include: [0423]
Proportion of subjects with an ADA-positive sample at baseline
[0424] Proportion of ADA-positive subjects (on treatment and
overall) [0425] Proportion of subjects who are persistently
positive (e.g., 2 or more sequential ADA-positive samples with an
adequate elapse of time in-between) [0426] Proportion of subjects
who have neutralizing antibodies detected in one or more
samples
[0427] Centrally Read ECGs (Parts A and B)
[0428] In Part A and Part B, QTc is assessed by a central reader at
follow-up visit 1, and on day 1 of Cycle 1 and Cycle 3 (pre-dose
and 4 hour post-dose time points). These assessments are used to
address the secondary objective of assessing the effect of
BMS-986016 administered alone and in combination with BMS-936558 on
QTc. ECGs assessed locally by the investigator are also collected
at the start of each cycle.
[0429] Biomarker Endpoints
[0430] Biomarker endpoints from peripheral blood are generally
measured at multiple timepoints, and evaluated as both predictive
and pharmacodynamic markers in the context of the exploratory
biomarker objectives. These may include measures such as levels and
change from baseline in levels of the following at each scheduled
timepoint: [0431] Serum soluble factors [0432] The proportion of
specific lymphocyte subsets/expression levels of T cell
co-stimulatory markers assessed using flow cytometry [0433]
Expression of genes encoding BMS-986016--stimulated effector
functions (perforin, granzyme B, and IFN-.gamma.) and genes
encoding T cell co-stimulatory receptors (PD-1, PD-L1, and CTLA-4).
[0434] Percent of subjects expressing single nucleotide
polymorphisms linked to PD-1 genes (per SNP) [0435] Measures of the
quantity and diversity of antibodies observed to tumor-associated
antigens (Part C only)
[0436] Biomarker endpoints from tumor biopsies are explored
predominantly in an effort to identify baseline markers predictive
of efficacy, since they are only measured at baseline for most
subjects. For the subset of subjects who have both pre-treatment
and on-treatment biopsies, pharmacodynamic associations are
explored. Endpoints may include measures, such as pre-treatment
levels and change in levels observed on-treatment of: [0437]
Functional status of lymphocytes measured as the percent of CD8+
T-cells positive for IFN-.gamma. and granzyme B expression, and the
geometric mean intensity (log-scale) of CD8+ cells that are
positive for IFN-.gamma. and granzyme B expression (via ex vivo
functional assay) [0438] Expression of genes encoding
BMS-986016-stimulated effector functions (perforin, granzyme B, and
IFN-.gamma.) and genes encoding T cell co-stimulatory receptors
(PD-1, PD-L1, and CTLA-4) [0439] IHC assessment of the
presence/absence and intensity (measured using a discrete scale:
such as 0, 1, 2, 3, 4) of expression of LAG-3, MHC class II, PD-1,
PD-L1, and PD-L2.
[0440] Appropriate functional transformation of these exploratory
measures are applied as necessary.
[0441] Pharmacokinetics
[0442] BMS-936558 concentration-time data at scheduled trough
(Ctrough) and end-of-infusion timepoints is evaluated as an
exploratory endpoint. Measurements are collected on treatment (up
to 12 cycles) and for up to 135 days during the post-treatment
follow-up.
[0443] PK parameters for BMS-986016 are calculated using
noncompartmental analyses. Summary statistics are tabulated for the
PK parameters of BMS-986016 by dose and study day/cycle. To
describe the association of these parameters with dose of
BMS-986016, scatter plots of Cmax and AUC(TAU) versus dose are
provided for each day/cycle measured. Dose proportionality of
BMS-986016 when administered alone or co-administered with
BMS-936558 is also assessed based on a power model. Trough
concentrations of BMS-986016 re plotted versus study day and cycle.
BMS-936558 end-of-infusion and trough (Ctrough) concentrations are
tabulated using summary statistics.
TABLE-US-00013 SEQUENCE SUMMARY SEQ ID NO: SEQUENCE 1 Heavy Chain
Amino Acid Sequence Anti-LAG-3 mAb (BMS-986016) (variable region
underlined; constant region bold)
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWIGE
INHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFGYS
DYEYNWFDPWGQGTLVTVSSASTKGPSVPPLAPCSRSTSESTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKT
YTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT
LPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS
DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK* 2 Light Chain
Amino Acid Sequence Anti-LAC-3 mAb (BMS-986016) (variable region
underlined; constant region bold)
EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQ
GTNLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV
DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC*
3 Heavy Chain Variable Region (VH) Amino Acid Sequence Anti-LAG-3
mAb (BMS-986016) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKGLEWI
GEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTAVYYCAFG
YSDYEYNWFDPWGQGTLVTVSS 4 Heavy Chain Variable Region (VH)
Nucleotide Sequence Anti-LAG-3 mAb (BMS-986016)
caggtgcagctacagcagtggggcgcaggactgttgaagccttcggagaccct
gtccctcacctgcgctgtctatggtgggtccttcagtgattactactggaact
ggatccgccagcccccagggaaggggctggagtggattggggaaatcaatcat
cgtggaagcaccaactccaacccgtccctcaagagtcgagtcaccctatcact
agacacgtccaagaaccagttctccctgaagctgaggtctgtgaccgccgcgg
acacggctgtgtattactgtgcgtttggatatagtgactacgagtacaactgg
ttcgacccctggggccagggaaccctggtcaccgtctcctca 5 Light Chain Variable
Region (VL) Amino Acid Sequence Anti-LAG-3 mAb (BMS-986016)
EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGQ GTNLEIK 6 Light
Chain Variable Region (VL) Nucleotide Sequence Anti-LAG-3 mAb
(BMS-986016) gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaag
agccaccctctcctgcagggccagtcagagtattagcagctacttagcctggt
accaacagaaacctggccaggctcccaggctcctcatctatgatgcatccaac
agggccactggcatcccagccaggttcagtggcagtgggtctgggacagactt
cactctcaccatcagcagcctagagcctgaagattttgcagtttattactgtc
agcagcgtagcaactggcctctcacttttggccaggggaccaacctggagatc aaa 7 Heavy
Chain CDR1 Amino Acid Sequence Anti-LAG-3 mAb (BMS-986016) DYYWN 8
Heavy Chain CDR2 Amino Acid Sequence Anti-LAG-3 mAb (BMS-986016)
EINHRGSTNSNPSLKS 9 Heavy Chain CDR3 Amino Acid Sequence Anti-LAG-3
mAb (BMS-986016) GYSDYEYNWFDP 10 Light Chain CDR1 Amino Acid
Sequence Anti-LAG-3 mAb (BMS-986016) RASQSISSYLA 11 Light Chain
CDR2 Amino Acid Sequence Anti-LAG-3 mAb (BMS-986016) DASNRAT 12
Light Chain CDR3 Amino Acid Sequence Anti-LAG-3 mAb (BMS-986016)
QQRSNWPLT 13 Human LAG-3 Amino Acid Sequence
MWEAQFLGLLFLQPLWVAPVKPLQPGAEVPVVWAQEGAPAQLPCSPTIPLQD
LSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVG
PGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDR
ALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQ
GRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLG
LEPPTPLIVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDN
GDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKL
LCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERL
LGAAVYFTELSSPGAQRSGRAPGALPAGHLLLFLTLGVLSLLLLVTGAFGFHLW
RRQWRPRRFSALEQGIHPPQAQSKIEELEQEPEPEPEPEPEPEPEPEPEQL* 14 LAG-3
Epitope PGHPLAPG 15 LAG-3 Epitope HPAAPSSW 16 LAG-3 Epitope
PAAPSSWG 17 Heavy Chain Amino Acid Sequence Anti-PD-1 mAb
(BM5936558; 5C4 in WO 2006/121168) (variable region underlined;
constant region bold) QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGL
EWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDT
AVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT
VPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLG
GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV
EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKG
LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPS
DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNV
FSCSVMHEALHNHYTQKSLSLSLGK 18 Light Chain Amino Acid Sequence
Anti-PD-1 mAb (BM5936558; 5C4 in WO 2006/121168) (variable region
underlined; constant region bold)
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI
YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPR
TFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA
KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHK
VYACEVTHQGLSSPVTKSFNRGEC 19 Heavy Chain Variable Region (VH) Amino
Acid Sequence Anti-PD-1 mAb (BMS936558; 5C4 in WO 2006/121168) (SEQ
ID NO: 4 from WO 2006/121168)
QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAV
IWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATND DYWGQGTLVTVSS 20
Heavy Chain Variable Region (VH) Nucleotide Sequence Anti-PD-1 mAb
(BMS936558; 5C4 in WO 2006/121168) (SEQ ID NO: 60 from WO
2006/121168) cag gtg cag ctg gtg gag tct ggg gga ggc gtg gtc cag
cct ggg agg tcc ctg aga ctc gac tgt aaa gcg tct gga atc acc ttc agt
aac tct ggc atg cac tgg gtc cgc cag gct cca ggc aag ggg ctg gag tgg
gtg gca gtt att tgg tat gat gga agt aaa aga tac tat gca gac tcc gtg
aag ggc cga ttc acc atc tcc aga gac aat tcc aag aac acg ctg ttt ctg
caa atg aac agc ctg aga gcc gag gac acg gct gtg tat tac tgt gcg aca
aac gac gac tac tgg ggc cag gga acc ctg gtc acc gtc tcc tca 21
Light Chain Variable Region (VL) Amino Acid Sequence Anti-PD-1 mAb
(BMS936558; 5C4 in WO 2006/121168) (SEQ ID NO: 11 from WO
2006/121168) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQ GTKVEIK 22 Light
Chain Variable Region (VL) Nucleotide Sequence Anti-PD-1 mAb
(BMS936558; 5C4 in WO 2006/121168) (SEQ ID NO: 67 from WO
2006/121168) gaa att gtg ttg aca cag tct cca gcc acc ctg tct ttg
tct cca ggg gaa aga gcc acc ctc tcc tgc agg gcc agt cag agt gtt agt
agt tac tta gcc tgg tac caa cag aaa cct ggc cag gct ccc agg ctc ctc
atc tat gat gca tcc aac agg gcc act ggc atc cca gcc agg ttc agt ggc
agt ggg tct ggg aca gac ttc act ctc acc atc agc agc cta gag cct gaa
gat ttt gca gtt tat tac tgt cag cag agt agc aac tgg cct cgg acg ttc
ggc caa ggg acc aag gtg gaa atc aaa 23 Heavy Chain CDR1 Amino Acid
Sequence Anti-PD-1 mAb (BMS936558; 5C4 in WO 2006/121168) (SEQ ID
NO: 18 from WO 2006/121168) NSGMH 24 Heavy Chain CDR2 Amino Acid
Sequence Anti-PD-1 mAb (BMS936558; 5C4 in WO 2006/121168) (SEQ ID
NO: 25 from WO 2006/121168) VIWYDGSKRYYADSVKG 25 Heavy Chain CDR3
Amino Acid Sequence Anti-PD-1 mAb (BMS936558; 5C4 in WO
2006/121168) (SEQ ID NO: 32 from WO 2006/121168) NDDY 26 Light
Chain CDR1 Amino Acid Sequence Anti-PD-1 mAb (BMS936558; 5C4 in WO
2006/121168) (SEQ ID NO: 39 from WO 2006/121168) RASQSVSSYLA 27
Light Chain CDR2 Amino Acid Sequence Anti-PD-1 mAb (BMS936558; 5C4
in WO 2006/121168) (SEQ ID NO: 46 from WO 2006/121168) DASNRAT 28
Light Chain CDR3 Amino Acid Sequence Anti-PD-1 mAb (BMS936558; 5C4
in WO 2006/121168) (SEQ ID NO: 53 from WO 2006/121168) QQSSNWPRT 29
Complete PD-1 sequence (GenBank Accession No.: U64863) agtttccctt
ccgctcacct ccgcctgagc agtggagaag gcggcactct ggtggggctg ctccaggcat
gcagatccca caggcgccct ggccagtcgt ctgggcggtg ctacaactgg gctggcggcc
aggatggttc ttagactccc cagacaggcc ctggaacccc cccaccttct tcccagccct
gctcgtggtg accgaagggg acaacgccac cttcacctgc agcttctcca acacatcgga
gagcttcgtg ctaaactggt accgcatgag ccccagcaac cagacggaca agctggccgc
cttccccgag gaccgcagcc agcccggcca ggactgccgc ttccgtgtca cacaactgcc
caacgggcgt gacttccaca tgagcgtggt cagggcccgg cgcaatgaca gcggcaccta
cctctgtggg gccatctccc tggcccccaa ggcgcagatc aaagagagcc tgcgggcaga
gctcagggtg acagagagaa gggcagaagt gcccacagcc caccccagcc cctcacccag
gccagccggc cagttccaaa ccctggtggt tggtgtcgtg ggcggcctgc tgggcagcct
ggtgctgcta gtctgggtcc tggccgtcat ctgctcccgg gccgcacgag ggacaatagg
agccaggcgc accggccagc ccctgaagga ggacccctca gccgtgcctg tgttctctgt
ggactatggg gagctggatt tccagtggcg agagaagacc ccggagcccc ccgtgccctg
tgtccctgag cagacggagt atgccaccat tgtctttcct agcggaatgg gcacctcatc
ccccgcccgc aggggctcag ccgacggccc tcggagtgcc cagccactga ggcctgagga
tggacactgc tcttggcccc tctgaccggc ttccttggcc accagtgttc tgcagaccct
ccaccatgag cccgggtcag cgcatttcct caggagaagc aggcagggtg caggccattg
caggccgtcc aggggctgag ctgcctgggg gcgaccgggg ctccagcctg cacctgcacc
aggcacagcc ccaccacagg actcatgtct caatgcccac agtgagccca ggcagcaggt
gtcaccgtcc cctacaggga gggccagatg cagtcactgc ttcaggtcct gccagcacag
agctgcctgc gtccagctcc ctgaatctct gctgctgctg ctgctgctgc tgctgctgcc
tgcggcccgg ggctgaaggc gccgtggccc tgcctgacgc cccggagcct cctgcctgaa
cttgggggct ggttggagat ggccttggag cagccaaggt gcccctggca gtggcatccc
gaaacgccct ggacgcaggg cccaagactg ggcacaggag tgggaggtac atggggctgg
ggactcccca ggagttatct gctccctgca ggcctagaga agtttcaggg aaggtcagaa
gagctcctgg ctgtggtggg cagggcagga aacccctccc acctttacac atgcccaggc
agcacctcag gccctttgtg gggcagggaa gctgaggcag taagcgggca ggcagagctg
gaggcctttc aggccagcca gcactctggc ctcctgccgc cgcattccac cccagcccct
cacaccactc gggagaggga catcctacgg tcccaaggtc aggagggcag ggctggggtt
gactcaggcc cctcccagct gtggccacct gggtgttggg agggcagaag tgcaggcacc
tagggccccc catgtgccca ccctgggagc tctccttgga acccattcct gaaattattt
aaaggggttg gccgggctcc caccagggcc tgggtgggaa ggtacaggcg ttcccccggg
gcctagtacc cccgcgtggc ctatccactc ctcacatcca cacactgcac ccccactcct
ggggcagggc caccagcatc caggcggcca gcaggcacct gagtggctgg gacaagggat
cccccttccc tgtggttcta ttatattata attataatta aatatgagag catgct Heavy
Chain Nucleotide Sequence Anti-LAG-3 mAb (BMS-986016)
caggtgcagctacagcagtggggcgcaggactgttgaagccttcggagaccctgtccct
cacctgcgctgtctatggtgggtccttcagtgattactactggaactggatccgccag
cccccagggaaggggctggagtggattggggaaatcaatcatcgtggaagcaccaact
ccaacccgtccctcaagagtcgagtcaccctatcactagacacgtccaagaaccagtt
ctccctgaagctgaggtctgtgaccgccgcggacacggctgtgtattactgtgcgtttg
gatatagtgactacgagtacaactggttcgacccctggggccagggaaccctggtcacc
gtctcctcagctagcaccaagggcccatccgtcttccccctggcgccctgctccaggagc
acctccgagagcacagccgccctgggctgcctggtcaaggactacttccccgaaccggtg
acggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtccta
cagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggc
acgaagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagaga
gttgagtccaaatatggtcccccatgcccaccatgcccagcacctgagttcctgggggga
ccatcagtcttcctgttccccccaaaacccaaggacactctcatgatctcccggacccct
gaggtcacgtgcgtggtggtggacgtgagccaggaagaccccgaggtccagttcaactgg
tacgtggatggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagttcaa
cagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaacggcaa
ggagtacaagtgcaaggtctccaacaaaggcctcccgtcctccatcgagaaaaccatctc
caaagccaaagggcagccccgagagccacaggtgtacaccctgcccccatcccaggagga
gatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacat
cgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgt
gctggactccgacggctccttcttcctctacagcaggctaaccgtggacaagagcaggtg
gcaggaggggaatgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacac
acagaagagcctctccctgtctctgggtaaatga Light Chain Nucleotide Sequence
Anti-LAG-3 mAb (BMS-986016)
gaaattgtgttgacacagtctccagccaccctgtctttgtctccaggggaaagagccacc
ctctcctgcagggccagtcagagtattagcagctacttagcctggtaccaacagaaacct
ggccaggctcccaggctcctcatctatgatgcatccaacagggccactggcatcccag
ccaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagcctagagc
ctgaagattttgcagtttattactgtcagcagcgtagcaactggcctctcacttttggcc
aggggaccaacctggagatcaaacgtacggtggctgcaccatctgtcttcatcttcccgc
catctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttct
atcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactccc
aggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctga
cgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagg
gcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag
Sequence CWU 1
1
321447PRTArtificial SequenceSynthetic Heavy Chain Amino Acid
Sequence; Anti-LAG-3 mAb (BMS-986016) 1Gln Val Gln Leu Gln Gln Trp
Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys
Ala Val Tyr Gly Gly Ser Phe Ser Asp Tyr 20 25 30Tyr Trp Asn Trp Ile
Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn
His Arg Gly Ser Thr Asn Ser Asn Pro Ser Leu Lys 50 55 60Ser Arg Val
Thr Leu Ser Leu Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys
Leu Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90
95Phe Gly Tyr Ser Asp Tyr Glu Tyr Asn Trp Phe Asp Pro Trp Gly Gln
100 105 110Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
Ser Val 115 120 125Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
Ser Thr Ala Ala 130 135 140Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro Val Thr Val Ser145 150 155 160Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr Phe Pro Ala Val 165 170 175Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190Ser Ser Ser
Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys 195 200 205Pro
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro 210 215
220Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser
Val225 230 235 240Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
Ile Ser Arg Thr 245 250 255Pro Glu Val Thr Cys Val Val Val Asp Val
Ser Gln Glu Asp Pro Glu 260 265 270Val Gln Phe Asn Trp Tyr Val Asp
Gly Val Glu Val His Asn Ala Lys 275 280 285Thr Lys Pro Arg Glu Glu
Gln Phe Asn Ser Thr Tyr Arg Val Val Ser 290 295 300Val Leu Thr Val
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys305 310 315 320Cys
Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile 325 330
335Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
Cys Leu 355 360 365Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
Trp Glu Ser Asn 370 375 380Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser385 390 395 400Asp Gly Ser Phe Phe Leu Tyr
Ser Arg Leu Thr Val Asp Lys Ser Arg 405 410 415Trp Gln Glu Gly Asn
Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 435 440
4452214PRTArtificial SequenceSynthetic Light Chain Amino Acid
Sequence; Anti-LAG-3 mAb (BMS-986016) 2Glu Ile Val Leu Thr Gln Ser
Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser
Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln
Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser
Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu 85 90
95Thr Phe Gly Gln Gly Thr Asn Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys
Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu
Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp
Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu
Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe
Asn Arg Gly Glu Cys 2103120PRTArtificial SequenceSynthetic Heavy
Chain Variable Region (VH) Amino Acid Sequence; Anti-LAG-3 mAb
(BMS-986016) 3Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys
Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser
Phe Ser Asp Tyr 20 25 30Tyr Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys
Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn His Arg Gly Ser Thr Asn
Ser Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Leu Ser Leu Asp Thr
Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Arg Ser Val Thr Ala
Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Phe Gly Tyr Ser Asp Tyr
Glu Tyr Asn Trp Phe Asp Pro Trp Gly Gln 100 105 110Gly Thr Leu Val
Thr Val Ser Ser 115 1204360DNAArtificial SequenceSynthetic Heavy
Chain Variable Region (VH) Nucleotide Sequence; Anti-LAG-3 mAb
(BMS-986016) 4caggtgcagc tacagcagtg gggcgcagga ctgttgaagc
cttcggagac cctgtccctc 60acctgcgctg tctatggtgg gtccttcagt gattactact
ggaactggat ccgccagccc 120ccagggaagg ggctggagtg gattggggaa
atcaatcatc gtggaagcac caactccaac 180ccgtccctca agagtcgagt
caccctatca ctagacacgt ccaagaacca gttctccctg 240aagctgaggt
ctgtgaccgc cgcggacacg gctgtgtatt actgtgcgtt tggatatagt
300gactacgagt acaactggtt cgacccctgg ggccagggaa ccctggtcac
cgtctcctca 3605107PRTArtificial SequenceSynthetic Light Chain
Variable Region (VL) Amino Acid Sequence; Anti-LAG-3 mAb
(BMS-986016) 5Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu
Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
Ile Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile
Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Arg Ser Asn Trp Pro Leu 85 90 95Thr Phe Gly Gln Gly Thr
Asn Leu Glu Ile Lys 100 1056321DNAArtificial SequenceSynthetic
Light Chain Variable Region (VL) Nucleotide Sequence; Anti-LAG-3
mAb (BMS-986016) 6gaaattgtgt tgacacagtc tccagccacc ctgtctttgt
ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtattagc agctacttag
cctggtacca acagaaacct 120ggccaggctc ccaggctcct catctatgat
gcatccaaca gggccactgg catcccagcc 180aggttcagtg gcagtgggtc
tgggacagac ttcactctca ccatcagcag cctagagcct 240gaagattttg
cagtttatta ctgtcagcag cgtagcaact ggcctctcac ttttggccag
300gggaccaacc tggagatcaa a 32175PRTArtificial SequenceSynthetic
Heavy Chain CDR1 Amino Acid Sequence; Anti-LAG-3 mAb (BMS-986016)
7Asp Tyr Tyr Trp Asn1 5816PRTArtificial SequenceSynthetic Heavy
Chain CDR2 Amino Acid Sequence; Anti-LAG-3 mAb (BMS-986016) 8Glu
Ile Asn His Arg Gly Ser Thr Asn Ser Asn Pro Ser Leu Lys Ser1 5 10
15912PRTArtificial SequenceSynthetic Heavy Chain CDR3 Amino Acid
Sequence; Anti-LAG-3 mAb (BMS-986016) 9Gly Tyr Ser Asp Tyr Glu Tyr
Asn Trp Phe Asp Pro1 5 101011PRTArtificial SequenceSynthetic Light
Chain CDR1 Amino Acid Sequence; Anti-LAG-3 mAb (BMS-986016) 10Arg
Ala Ser Gln Ser Ile Ser Ser Tyr Leu Ala1 5 10117PRTArtificial
SequenceSynthetic Light Chain CDR2 Amino Acid Sequence; Anti-LAG-3
mAb (BMS-986016) 11Asp Ala Ser Asn Arg Ala Thr1 5129PRTArtificial
SequenceSynthetic Light Chain CDR3 Amino Acid Sequence; Anti-LAG-3
mAb (BMS-986016) 12Gln Gln Arg Ser Asn Trp Pro Leu Thr1
513525PRTHomo sapiensmisc_featureHuman LAG-3 Amino Acid Sequence
13Met Trp Glu Ala Gln Phe Leu Gly Leu Leu Phe Leu Gln Pro Leu Trp1
5 10 15Val Ala Pro Val Lys Pro Leu Gln Pro Gly Ala Glu Val Pro Val
Val 20 25 30Trp Ala Gln Glu Gly Ala Pro Ala Gln Leu Pro Cys Ser Pro
Thr Ile 35 40 45Pro Leu Gln Asp Leu Ser Leu Leu Arg Arg Ala Gly Val
Thr Trp Gln 50 55 60His Gln Pro Asp Ser Gly Pro Pro Ala Ala Ala Pro
Gly His Pro Leu65 70 75 80Ala Pro Gly Pro His Pro Ala Ala Pro Ser
Ser Trp Gly Pro Arg Pro 85 90 95Arg Arg Tyr Thr Val Leu Ser Val Gly
Pro Gly Gly Leu Arg Ser Gly 100 105 110Arg Leu Pro Leu Gln Pro Arg
Val Gln Leu Asp Glu Arg Gly Arg Gln 115 120 125Arg Gly Asp Phe Ser
Leu Trp Leu Arg Pro Ala Arg Arg Ala Asp Ala 130 135 140Gly Glu Tyr
Arg Ala Ala Val His Leu Arg Asp Arg Ala Leu Ser Cys145 150 155
160Arg Leu Arg Leu Arg Leu Gly Gln Ala Ser Met Thr Ala Ser Pro Pro
165 170 175Gly Ser Leu Arg Ala Ser Asp Trp Val Ile Leu Asn Cys Ser
Phe Ser 180 185 190Arg Pro Asp Arg Pro Ala Ser Val His Trp Phe Arg
Asn Arg Gly Gln 195 200 205Gly Arg Val Pro Val Arg Glu Ser Pro His
His His Leu Ala Glu Ser 210 215 220Phe Leu Phe Leu Pro Gln Val Ser
Pro Met Asp Ser Gly Pro Trp Gly225 230 235 240Cys Ile Leu Thr Tyr
Arg Asp Gly Phe Asn Val Ser Ile Met Tyr Asn 245 250 255Leu Thr Val
Leu Gly Leu Glu Pro Pro Thr Pro Leu Thr Val Tyr Ala 260 265 270Gly
Ala Gly Ser Arg Val Gly Leu Pro Cys Arg Leu Pro Ala Gly Val 275 280
285Gly Thr Arg Ser Phe Leu Thr Ala Lys Trp Thr Pro Pro Gly Gly Gly
290 295 300Pro Asp Leu Leu Val Thr Gly Asp Asn Gly Asp Phe Thr Leu
Arg Leu305 310 315 320Glu Asp Val Ser Gln Ala Gln Ala Gly Thr Tyr
Thr Cys His Ile His 325 330 335Leu Gln Glu Gln Gln Leu Asn Ala Thr
Val Thr Leu Ala Ile Ile Thr 340 345 350Val Thr Pro Lys Ser Phe Gly
Ser Pro Gly Ser Leu Gly Lys Leu Leu 355 360 365Cys Glu Val Thr Pro
Val Ser Gly Gln Glu Arg Phe Val Trp Ser Ser 370 375 380Leu Asp Thr
Pro Ser Gln Arg Ser Phe Ser Gly Pro Trp Leu Glu Ala385 390 395
400Gln Glu Ala Gln Leu Leu Ser Gln Pro Trp Gln Cys Gln Leu Tyr Gln
405 410 415Gly Glu Arg Leu Leu Gly Ala Ala Val Tyr Phe Thr Glu Leu
Ser Ser 420 425 430Pro Gly Ala Gln Arg Ser Gly Arg Ala Pro Gly Ala
Leu Pro Ala Gly 435 440 445His Leu Leu Leu Phe Leu Thr Leu Gly Val
Leu Ser Leu Leu Leu Leu 450 455 460Val Thr Gly Ala Phe Gly Phe His
Leu Trp Arg Arg Gln Trp Arg Pro465 470 475 480Arg Arg Phe Ser Ala
Leu Glu Gln Gly Ile His Pro Pro Gln Ala Gln 485 490 495Ser Lys Ile
Glu Glu Leu Glu Gln Glu Pro Glu Pro Glu Pro Glu Pro 500 505 510Glu
Pro Glu Pro Glu Pro Glu Pro Glu Pro Glu Gln Leu 515 520
525148PRTArtificial SequenceSynthetic LAG-3 Epitope 14Pro Gly His
Pro Leu Ala Pro Gly1 5158PRTArtificial SequenceSynthetic LAG-3
Epitope 15His Pro Ala Ala Pro Ser Ser Trp1 5168PRTArtificial
SequenceSynthetic LAG-3 Epitope 16Pro Ala Ala Pro Ser Ser Trp Gly1
517440PRTArtificial SequenceSynthetic Heavy Chain Amino Acid
Sequence; Anti-PD-1 mAb (BMS936558) 17Gln Val Gln Leu Val Glu Ser
Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Asp Cys
Lys Ala Ser Gly Ile Thr Phe Ser Asn Ser 20 25 30Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp
Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe65 70 75 80Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser
100 105 110Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
Cys Ser 115 120 125Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
Leu Val Lys Asp 130 135 140Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn Ser Gly Ala Leu Thr145 150 155 160Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln Ser Ser Gly Leu Tyr 165 170 175Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys 180 185 190Thr Tyr Thr
Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp 195 200 205Lys
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala 210 215
220Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro225 230 235 240Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
Thr Cys Val Val 245 250 255Val Asp Val Ser Gln Glu Asp Pro Glu Val
Gln Phe Asn Trp Tyr Val 260 265 270Asp Gly Val Glu Val His Asn Ala
Lys Thr Lys Pro Arg Glu Glu Gln 275 280 285Phe Asn Ser Thr Tyr Arg
Val Val Ser Val Leu Thr Val Leu His Gln 290 295 300Asp Trp Leu Asn
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly305 310 315 320Leu
Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 325 330
335Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr
340 345 350Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
Pro Ser 355 360 365Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
Glu Asn Asn Tyr 370 375 380Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr385 390 395 400Ser Arg Leu Thr Val Asp Lys
Ser Arg Trp Gln Glu Gly Asn Val Phe 405 410 415Ser Cys Ser Val Met
His Glu Ala Leu His Asn His Tyr Thr Gln Lys 420 425 430Ser Leu Ser
Leu Ser Leu Gly Lys 435 44018214PRTArtificial SequenceSynthetic
Light Chain Amino Acid Sequence; Anti-PD-1 mAb (BMS936558) 18Glu
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
Ile 35 40 45Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser
Ser Asn Trp Pro Arg 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165
170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
Tyr 180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys
21019113PRTArtificial SequenceSynthetic Heavy Chain Variable Region
(VH) Amino Acid Sequence; Anti-PD-1 mAb (BMS936558) 19Gln Val Gln
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu
Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr Phe Ser Asn Ser 20 25 30Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val
50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Phe65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser 100 105 110Ser20339DNAArtificial SequenceSynthetic
Heavy Chain Variable Region (VH) Nucleotide Sequence; Anti-PD-1 mAb
(BMS936558) 20caggtgcagc tggtggagtc tgggggaggc gtggtccagc
ctgggaggtc cctgagactc 60gactgtaaag cgtctggaat caccttcagt aactctggca
tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt
atttggtatg atggaagtaa aagatactat 180gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacgctgttt 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gacaaacgac
300gactactggg gccagggaac cctggtcacc gtctcctca 33921107PRTArtificial
SequenceSynthetic Light Chain Variable Region (VL) Amino Acid
Sequence; Anti-PD-1 mAb (BMS936558) 21Glu Ile Val Leu Thr Gln Ser
Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser
Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln
Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser
Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Asn Trp Pro Arg 85 90
95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100
10522321DNAArtificial SequenceSynthetic Light Chain Variable Region
(VL) Nucleotide Sequence; Anti-PD-1 mAb (BMS936558) 22gaaattgtgt
tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60ctctcctgca
gggccagtca gagtgttagt agttacttag cctggtacca acagaaacct
120ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg
catcccagcc 180aggttcagtg gcagtgggtc tgggacagac ttcactctca
ccatcagcag cctagagcct 240gaagattttg cagtttatta ctgtcagcag
agtagcaact ggcctcggac gttcggccaa 300gggaccaagg tggaaatcaa a
321235PRTArtificial SequenceSynthetic Heavy Chain CDR1 Amino Acid
Sequence; Anti-PD-1 mAb (BMS936558) 23Asn Ser Gly Met His1
52417PRTArtificial SequenceSynthetic Heavy Chain CDR2 Amino Acid
Sequence; Anti-PD-1 mAb (BMS936558) 24Val Ile Trp Tyr Asp Gly Ser
Lys Arg Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly254PRTArtificial
SequenceSynthetic Heavy Chain CDR3 Amino Acid Sequence; Anti-PD-1
mAb (BMS936558) 25Asn Asp Asp Tyr12611PRTArtificial
SequenceSynthetic Light Chain CDR1 Amino Acid Sequence; Anti-PD-1
mAb (BMS936558) 26Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala1 5
10277PRTArtificial SequenceSynthetic Light Chain CDR2 Amino Acid
Sequence; Anti-PD-1 mAb (BMS936558) 27Asp Ala Ser Asn Arg Ala Thr1
5289PRTArtificial SequenceSynthetic Light Chain CDR3 Amino Acid
Sequence; Anti-PD-1 mAb (BMS936558) 28Gln Gln Ser Ser Asn Trp Pro
Arg Thr1 5292106DNAHomo sapiensmisc_featureComplete PD-1 sequence
29agtttccctt ccgctcacct ccgcctgagc agtggagaag gcggcactct ggtggggctg
60ctccaggcat gcagatccca caggcgccct ggccagtcgt ctgggcggtg ctacaactgg
120gctggcggcc aggatggttc ttagactccc cagacaggcc ctggaacccc
cccaccttct 180tcccagccct gctcgtggtg accgaagggg acaacgccac
cttcacctgc agcttctcca 240acacatcgga gagcttcgtg ctaaactggt
accgcatgag ccccagcaac cagacggaca 300agctggccgc cttccccgag
gaccgcagcc agcccggcca ggactgccgc ttccgtgtca 360cacaactgcc
caacgggcgt gacttccaca tgagcgtggt cagggcccgg cgcaatgaca
420gcggcaccta cctctgtggg gccatctccc tggcccccaa ggcgcagatc
aaagagagcc 480tgcgggcaga gctcagggtg acagagagaa gggcagaagt
gcccacagcc caccccagcc 540cctcacccag gccagccggc cagttccaaa
ccctggtggt tggtgtcgtg ggcggcctgc 600tgggcagcct ggtgctgcta
gtctgggtcc tggccgtcat ctgctcccgg gccgcacgag 660ggacaatagg
agccaggcgc accggccagc ccctgaagga ggacccctca gccgtgcctg
720tgttctctgt ggactatggg gagctggatt tccagtggcg agagaagacc
ccggagcccc 780ccgtgccctg tgtccctgag cagacggagt atgccaccat
tgtctttcct agcggaatgg 840gcacctcatc ccccgcccgc aggggctcag
ccgacggccc tcggagtgcc cagccactga 900ggcctgagga tggacactgc
tcttggcccc tctgaccggc ttccttggcc accagtgttc 960tgcagaccct
ccaccatgag cccgggtcag cgcatttcct caggagaagc aggcagggtg
1020caggccattg caggccgtcc aggggctgag ctgcctgggg gcgaccgggg
ctccagcctg 1080cacctgcacc aggcacagcc ccaccacagg actcatgtct
caatgcccac agtgagccca 1140ggcagcaggt gtcaccgtcc cctacaggga
gggccagatg cagtcactgc ttcaggtcct 1200gccagcacag agctgcctgc
gtccagctcc ctgaatctct gctgctgctg ctgctgctgc 1260tgctgctgcc
tgcggcccgg ggctgaaggc gccgtggccc tgcctgacgc cccggagcct
1320cctgcctgaa cttgggggct ggttggagat ggccttggag cagccaaggt
gcccctggca 1380gtggcatccc gaaacgccct ggacgcaggg cccaagactg
ggcacaggag tgggaggtac 1440atggggctgg ggactcccca ggagttatct
gctccctgca ggcctagaga agtttcaggg 1500aaggtcagaa gagctcctgg
ctgtggtggg cagggcagga aacccctccc acctttacac 1560atgcccaggc
agcacctcag gccctttgtg gggcagggaa gctgaggcag taagcgggca
1620ggcagagctg gaggcctttc aggccagcca gcactctggc ctcctgccgc
cgcattccac 1680cccagcccct cacaccactc gggagaggga catcctacgg
tcccaaggtc aggagggcag 1740ggctggggtt gactcaggcc cctcccagct
gtggccacct gggtgttggg agggcagaag 1800tgcaggcacc tagggccccc
catgtgccca ccctgggagc tctccttgga acccattcct 1860gaaattattt
aaaggggttg gccgggctcc caccagggcc tgggtgggaa ggtacaggcg
1920ttcccccggg gcctagtacc cccgcgtggc ctatccactc ctcacatcca
cacactgcac 1980ccccactcct ggggcagggc caccagcatc caggcggcca
gcaggcacct gagtggctgg 2040gacaagggat cccccttccc tgtggttcta
ttatattata attataatta aatatgagag 2100catgct 2106301344DNAArtificial
SequenceSynthetic Heavy Chain Nucleotide Sequence; Anti-LAG-3 mAb
(BMS-986016) 30caggtgcagc tacagcagtg gggcgcagga ctgttgaagc
cttcggagac cctgtccctc 60acctgcgctg tctatggtgg gtccttcagt gattactact
ggaactggat ccgccagccc 120ccagggaagg ggctggagtg gattggggaa
atcaatcatc gtggaagcac caactccaac 180ccgtccctca agagtcgagt
caccctatca ctagacacgt ccaagaacca gttctccctg 240aagctgaggt
ctgtgaccgc cgcggacacg gctgtgtatt actgtgcgtt tggatatagt
300gactacgagt acaactggtt cgacccctgg ggccagggaa ccctggtcac
cgtctcctca 360gctagcacca agggcccatc cgtcttcccc ctggcgccct
gctccaggag cacctccgag 420agcacagccg ccctgggctg cctggtcaag
gactacttcc ccgaaccggt gacggtgtcg 480tggaactcag gcgccctgac
cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540ggactctact
ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacgaagacc
600tacacctgca acgtagatca caagcccagc aacaccaagg tggacaagag
agttgagtcc 660aaatatggtc ccccatgccc accatgccca gcacctgagt
tcctgggggg accatcagtc 720ttcctgttcc ccccaaaacc caaggacact
ctcatgatct cccggacccc tgaggtcacg 780tgcgtggtgg tggacgtgag
ccaggaagac cccgaggtcc agttcaactg gtacgtggat 840ggcgtggagg
tgcataatgc caagacaaag ccgcgggagg agcagttcaa cagcacgtac
900cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaacggcaa
ggagtacaag 960tgcaaggtct ccaacaaagg cctcccgtcc tccatcgaga
aaaccatctc caaagccaaa 1020gggcagcccc gagagccaca ggtgtacacc
ctgcccccat cccaggagga gatgaccaag 1080aaccaggtca gcctgacctg
cctggtcaaa ggcttctacc ccagcgacat cgccgtggag 1140tgggagagca
atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc
1200gacggctcct tcttcctcta cagcaggcta accgtggaca agagcaggtg
gcaggagggg 1260aatgtcttct catgctccgt gatgcatgag gctctgcaca
accactacac acagaagagc 1320ctctccctgt ctctgggtaa atga
134431645DNAArtificial SequenceSynthetic Light Chain Nucleotide
Sequence; Anti-LAG-3 mAb (BMS-986016) 31gaaattgtgt tgacacagtc
tccagccacc ctgtctttgt ctccagggga aagagccacc 60ctctcctgca gggccagtca
gagtattagc agctacttag cctggtacca acagaaacct 120ggccaggctc
ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc
180aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag
cctagagcct 240gaagattttg cagtttatta ctgtcagcag cgtagcaact
ggcctctcac ttttggccag 300gggaccaacc tggagatcaa acgtacggtg
gctgcaccat ctgtcttcat cttcccgcca 360tctgatgagc agttgaaatc
tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420cccagagagg
ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag
480gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag
caccctgacg 540ctgagcaaag cagactacga gaaacacaaa gtctacgcct
gcgaagtcac ccatcagggc 600ctgagctcgc ccgtcacaaa gagcttcaac
aggggagagt gttag 645326PRTArtificial SequenceSynthetic Motif 32Met
Tyr Pro Pro Pro Tyr1 5
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