Note: Descriptions are shown in the official language in which they were submitted.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
1
METHODS OF TREATING SYSTEMIC LUPUS ERYTHEMATOSUS, SCLERODERMA,
AND MYOSITIS WITH AN ANTIBODY AGAINST INTERFERON-ALPHA
Inventors: Ryan Criste
Lorin Roskos
Wendy White
Rajesh Narwal
Dominique Ethgen
Gabriel Robbie
BACKGROUND OF THE DISCLOSURE
FIELD OF THE DISCLOSURE
[0001] The
present disclosure provides methods for treatment of autoimmune
diseases such as systemic lupus erythematosus, scleroderma, and myositis with
anti-IFN-alpha antibodies.
BACKGROUND ART
[0002] Type
I interferons (IFN) (IFN-alpha, IFN-beta, IFN-omega, IFN-tau) are a
family of structurally related cytokines having antiviral, antitumor and
immunomodulatory effects (Hardy et al. (2001) Blood 97:473; Cutrone and
Langer (2001) J. Biol. Chem. 276:17140). The human IFN-alpha locus includes
two subfamilies. The first subfamily consists of at least 14 non allelic genes
and 4
pseudogenes having at least 75% homology. The second subfamily, alpha-II or
omega, contains 5 pseudogenes and 1 functional gene which exhibits 70%
homology with the IFN-alpha genes. The subtypes of IFN-alpha have different
specific activities but they possess the same biological spectrum (Streuli et
al.
(1981) Proc. Natl. Acad. Sci. USA 78:2848) and have the same cellular receptor
(Agnet M. et al. (1983) in "Interferon 5" Ed. I. Gresser p. 1-22, Academic
Press,
London).
[0003] Increased expression of type I interferons has been described in
numerous autoimmune diseases (Foulis et al. (1987) Lancet 2:1423; Hooks et
al. (1982) Arthritis Rheum 25:396; Hertzog et
al. (1988) Clin. lmmunol.
lmmunopathol. 48:192; Hopkins and Meager (1988) Clin. Exp. lmmunol. 73:88;
Arvin and Miller (1984) Arthritis Rheum. 27:582). The most studied examples of
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
2
this are insulin-dependent diabetes mellitus (IDDM) (Foulis (1987) supra),
systemic lupus erythematosus (SLE) (Hooks (1982) supra; Blanco et al. (2001)
Science 294:1540; Ytterberg and Schnitzer (1982) Arthritis Rheum. 25:401;
Batteux et al. (1999) Eur. Cytokine Netw.:509), and autoimmune thyroiditis
(Prummel and Laurberg (2003) Thyroid 13:547; Mazziotti et al. (2002) J.
Endocrinol. Invest. 25:624; You et al. (1999) Chin. Med. J. 112:61; Koh et al.
(1997) Thyroid 7:891), which are all associated with elevated levels of IFN a,
and
rheumatoid arthritis (RA) (Hertzog (1988), Hopkins and Meager (1988), Arvin
and
Miller (1984), supra) in which IFN-beta may play a more significant role. For
a
review on the role of IFN-alpha on organ-targeted autoimmune and inflammatory
diseases see Crow (2010) Arthritis Res. Ther. 12:S5.
[0004] Systemic Lupus Erythematosus (SLE), often referred to as lupus, is a
prototypic systemic autoimmune disease. The disease includes constitutional
symptoms and signs, musculoskeletal, cutaneous, renal, gastrointestinal,
pulmonary, cardiac, reticuloendothelial, hematological and neuropsychiatric
manifestations. The cutaneous manifestations are among the most common in
SLE. A substantial body of evidence suggests type I interferons (IFNs),
particularly IFN-alpha, play a key role in the pathogenesis of systemic lupus
erythematosus (SLE). For a review on the use of anticytokine therapy in the
treatment of SLE see Yoo (2010) Lupus 19:1460.
[0005] Scleroderma, or systemic sclerosis (SSC), is a progressive,
debilitating
autoimmune disorder characterized by excess protein deposition into the
extracellular matrix by dermal fibroblasts, also referred to as dermal
fibrosis.
Patients with diffuse cutaneous disease often present unique markers such as
upregulation of type I interferon (IFN)-induced genes in skin. Supporting the
idea
that IFN plays a role in dermal fibrosis are recent reports of scleroderma
arising
in patients receiving IFN therapy for chronic viral infection. For a review of
systemic sclerosis, see Varga & Abraham (2007) J. Clin. Invest. 117:557-567.
See also Coelho et al. (2008) lmmunol. Lett. 118:110-115.
[0006] Myositis, a general term for inflammation of the muscles, is a group
of
conditions that are frequently associated with autoimmune conditions. Types of
myositis include, e.g., myositis ossificans, fibromyositis, (idiopathic)
inflammatory
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
3
myopathies, dermatomyositis, juvenile dermatomyositis, polymyositis, inclusion
body myositis, and pyomyositis
[0007] Administration of IFN-alpha has been reported to exacerbate
underlying
disease in patients with psoriasis, autoimmune thyroiditis and multiple
sclerosis
and to induce an SLE like syndrome in patients without a previous history of
autoimmune disease. Interferon a has also been shown to induce
glomerulonephritis in normal mice and to accelerate the onset of the
spontaneous
autoimmune disease of NZB/W mice. Further, IFN-alpha therapy has been
shown in some cases to lead to undesired side effects, including fever and
neurological disorders. Thus, there are pathological situations in which
inhibition
of IFN-alpha activity may be beneficial to the patient and a need exists for
methods of treatment effective in inhibiting IFN-alpha activity.
BRIEF SUMMARY OF THE DISCLOSURE
[0008] The disclosure provides methods of treating autoimmune diseases such
as SLE, SSC, and myositis in a human subject comprising administration of an
anti-interferon alpha antibody. These methods can be used for therapeutic,
including prophylactic, purposes, for example in situations where the
production
or expression of interferon alpha is associated with pathological symptoms. In
some embodiments sifalimumab (MEDI-545), an investigational fully human IgG1
monoclonal antibody against interferon-alpha, is used.
[0009] In one embodiment, the disclosure provides a method of treating an
autoimmune disorder in a human subject comprising administering to the subject
an antibody, or antigen-binding fragment thereof, which specifically binds to
human interferon alpha, wherein one or more pharmacokinetic characteristics
chosen from a clearance rate (CL, CLõ, CL/F, or CL/F) of between about 99
and about 432 mL/day, an apparent volume of distribution (Võ or Vz/F) of
between about 3 and about 17 L, and a serum half-life of about 14 days to
about
48 days is achieved following the administration; and wherein the autoimmune
disorder is systemic lupus erythematosus, scleroderma, or myositis.
[0010] In some embodiments, the antibody or antigen binding fragment
thereof
binds an epitope on human interferon alpha recognized by an antibody
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
4
comprising a heavy chain variable region comprising the amino acid sequence of
SEQ ID NO: 19 and a light chain variable region comprising the amino acid
sequence of SEQ ID NO: 22. In other embodiments, the antibody or antigen
binding fragment thereof comprises: (a) a heavy chain variable region CDR1
comprising SEQ ID NO: 1; (b) a heavy chain variable region CDR2 comprising
SEQ ID NO: 4; (c) a heavy chain variable region CDR3 comprising SEQ ID NO:
7; (d) a light chain variable region CDR1 comprising SEQ ID NO: 10; (e) a
light
chain variable region CDR2 comprising SEQ ID NO: 13; and (f) a light chain
variable region CDR3 comprising SEQ ID NO: 16. In some other embodiments,
the antibody or antigen binding fragment thereof comprises: (a) a heavy chain
variable region comprising the amino acid sequence of SEQ ID NO: 19, SEQ ID
NO:34, SEQ ID NO; 35, SEQ ID NO:36 or SEQ ID NO:37; and (b) a light chain
variable region comprising the amino acid sequence of SEQ ID NO: 22.
[0011] The antibody or antigen binding fragment thereof can be a human
antibody, a chimeric antibody, a humanized antibody, or an antigen binding
fragment thereof. In some embodiments, the antibody or antigen binding
fragment thereof is an IgG1 or IgG4 antibody or antigen-binding fragment
thereof.
In certain embodiments, the antigen binding fragment is a Fab antibody
fragment
or a single chain antibody (scFv).
[0012] In some embodiments, the antibody or antigen-binding fragment
thereof is
administered in a dosage dependent on the subject's body weight. In certain
embodiments, such weight-based dosage ranges from about 0.01 mg/kg to about
100 mg/kg of the subject's body weight. In some embodiments, such weight-
based dosage is chosen from about 0.3 mg/kg body weight, about 1 mg/kg body
weight, about 3 mg/kg body weight, and about 10 mg/kg body weight.
[0013] In other embodiments, the antibody or antigen-binding fragment
thereof is
administered as a fixed dosage. In certain embodiments, such fixed dosage
ranges from about 50 mg to about 2000 mg. In other embodiments, the fixed
dosage is chosen from about 100 mg, about 200 mg, about 600 mg, and about
1200 mg.
[0014] In some embodiments, the antibody or antigen-binding fragment
thereof is
administered as a single dose or is administered in two or more doses once per
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
week, once every two weeks, once every three weeks, once every four weeks,
once a month, once every 3 months, once every six months, or at varying
intervals. In some embodiments, a loading dose is administered at Day 14.
[0015] In certain embodiments, the administration is by a route chosen from
intravenous, intramuscular, intraperitoneal, intracerobrospinal, subcutaneous,
intra-articular, intrasynovial, intrathecal, oral, topical, inhalation, and a
combination of two or more recited routes.
[0016] In some embodiments, the administration is intravenous (IV)
administration. In some embodiments, IV administration is by IV infusion over
a
period of time. In some embodiments, the time to reach maximum plasma
concentration (T. or Tmax ss) following IV administration is about 0.13 days
or
less. In some embodiments, a single IV administration of about 0.3 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tmax of
about 0.12 days or less, a maximum plasma concentration (C.) of about 7 to
about 15 pg/mL, an area under the plasma concentration-time curve during a
dosage interval (T) (AUCT) of about 54 to about 104 pg day/mL, and a trough
plasma concentration (Ctrough) of about 2 to about 4 pg/mL. In other
embodiments,
a single IV administration of about 0.3 mg/kg to a population of subjects
achieves
one or more pharmacokinetic characteristics chosen from: an average T. of
about 0.07 days, an average C. of about 11 pg/mL, an average AUCT of about
79 pg day/mL, and an average Ctrough of about 3 pg/mL.
[0017] In some embodiments, a single IV administration of about 1 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tmax of
about 0.12 days or less, a C. of about 21 to about 43 pg/mL, an AUCT of about
153 to about 290 pg day/mL, and a Ctrough of about 4 to about 12 pg/mL. In
some
embodiments, a single IV administration of about 1 mg/kg to a population of
subjects achieves one or more pharmacokinetic characteristics chosen from: an
average T. of about 0.08 days, an average C. of about 32 pg/mL, an average
AUCT of about 221 pg day/mL, and an average Ctrough of about 8 pg/mL.
[0018] In some embodiments, a single IV administration of about 3 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tmax of
about 0.13 days or less, a C. of about 64 to about 143 pg/mL, an AUCT of
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
6
about 469 to about 1010 pg day/mL, and a Ctrough of about 12 to about 35
pg/mL.
In other embodiments, a single IV administration of about 3 mg/kg to a
population
of subjects achieves one or more pharmacokinetic characteristics chosen from:
an average Tmax of about 0.09 days, an average Cmax of about 103 pg/mL, an
average AUCT of about 739 pg day/mL, and an average Ctrough of about 23 pg/mL.
[0019] In some embodiments, a single IV administration of about 10 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tmax of
about 0.13 days or less, a Cmax of about 141 to about 318 pg/mL, an AUCT of
about 979 to about 2241 pg day/mL, and a Ctrough of about 27 to about 76
pg/mL.
In other embodiments, a single IV administration of about 10 mg/kg to a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from: an average Tmax of about 0.09 days, an average Cmax of about 230
pg/mL, an average AUCT of about 1610 pg day/mL, and an average Ctrough of
about 52 pg/mL.
[0020] In some embodiments, the administration of a sufficient number of IV
doses of about 0.3 mg/kg at about 14-day intervals achieves a steady state,
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
Tmax ss of about 0.60 days or less, a Cmax ss of about 11 to about 25 pg/mL,
an
AUCT ss of about 89 to about 197 pg day/mL, and a Ctrough ss of about 5 to
about
11 pg/mL is achieved.
[0021] In some embodiments, a sufficient number of IV doses of about 0.3
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, wherein one or more pharmacokinetic characteristics chosen
from: an average Tmax ss of about 0.17 days, an average Cmax ss of about 18
pg/mL, an average AUCT ss of about 143 pg day/mL, and an average Ctrough ss of
about 8 pg/mL is achieved.
[0022] In some embodiments, a sufficient number of IV doses of about 0.3
mg/kg
are administered at about 14-day intervals to a achieve a steady state,
wherein
one or more pharmacokinetic characteristics chosen from a clearance rate
(CLss)
of between about 99 and about 271 mL/day, an apparent volume of distribution
(Vss) of between about 4 and about 9 L, and a serum half-life of about 15 days
to
about 43 days is achieved.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
7
[0023] In certain embodiments, a sufficient number of IV doses of about 0.3
mg/kg are administered to a population of subjects at about 14-day intervals
to
achieve a steady state, wherein one or more pharmacokinetic characteristics
chosen from an average clearance rate (CLõ) of about 185 mL/day, an average
apparent volume of distribution (Vss) of about 6 L, and an average serum half-
life
of about 29 days is achieved.
[0024] In some embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered at about 14-day intervals to achieve a steady state, wherein
one
or more steady state pharmacokinetic characteristics chosen from: a Tmax ss of
about 0.11 days or less, a C max ss of about 29 to about 67 pg/mL, an AUCT ss
of
about 213 to about 591 pg day/mL, and a Ctrough ss of about 9 to about 30
pg/mL is
achieved.
[0025] In other embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, wherein one or more pharmacokinetic characteristics chosen
from: an average Tmax ss of about 0.07 days, an average Cmax ss of about 48
pg/mL, an average AUCT ss of about 197 pg day/mL, and an average Ctrough ss of
about 11 pg/mL is achieved.
[0026] In some embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered at about 14-day intervals to a achieve a steady state,
wherein
one or more pharmacokinetic characteristics chosen from a clearance rate (CL)
of between about 118 and about 348 mL/day, an apparent volume of distribution
(Vss) of between about 4 and about 9 L, and a serum half-life of about 15 days
to
about 32 days is achieved.
[0027] In some embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, wherein one or more pharmacokinetic characteristics chosen
from
an average clearance rate (CLss) of about 233 mL/day, an average apparent
volume of distribution (Vss) of about 6 L, and an average serum half-life of
about
23 days is achieved.
[0028] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered at about 14-day intervals to achieve a steady state, wherein
one
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
8
or more steady state pharmacokinetic characteristics chosen from: a T. õ of
about 0.33 days or less, a Cmax ss of about 75 to about 232 pg/mL, an AUCT ss
of
about 533 to about 1843 pg day/mL, and a Ctrough ss of about 26 to about 74
pg/mL is achieved.
[0029] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, wherein one or more pharmacokinetic characteristics chosen
from: an average Tmax ss of about 0.13 days, an average Cmax ss of about 153
pg/mL, an average AUCT ss of about 1188 pg day/mL, and an average Ctrough ss
of
about 50 pg/mL is achieved.
[0030] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered at about 14-day intervals to a achieve a steady state,
wherein
one or more pharmacokinetic characteristics chosen from a clearance rate
(CLss)
of between about 136 and about 304 mL/day, an apparent volume of distribution
(Vss) of between about 3 and about 7 L, and a serum half-life of about 14 days
to
about 26 days is achieved.
[0031] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, wherein one or more pharmacokinetic characteristics chosen
from
an average clearance rate (CLss) of about 220 mL/day, an average apparent
volume of distribution (Vss) of about 5 L, and an average serum half-life of
about
20 days is achieved.
[0032] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered at about 14-day intervals to achieve a steady state, wherein
one
or more steady state pharmacokinetic characteristics chosen from: a T. ss
about
0.82 days or less, a Cmax ss of about 288 to about 595 pg/mL, an AUCT ss of
about
2539 to about 4267 pg day/mL, and a Ctrough ss of about 93 to about 275 pg/mL
is
achieved.
[0033] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, wherein one or more pharmacokinetic characteristics chosen
from: an average T. ss of about 0.23 days, an average C. ss of about 232
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
9
pg/mL, an average AUG, ss of about 3403 pg day/mL, and an average Ctrough ss
of
about 184 pg/mL is achieved.
[0034] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered at about 14-day intervals to a achieve a steady state,
wherein
one or more pharmacokinetic characteristics chosen from a clearance rate (CL)
of between about 157 and about 319 mL/day, an apparent volume of distribution
(Vss) of between about 4 and about 7 L, and a serum half-life of about 15 days
to
about 29 days is achieved.
[0035] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics chosen
from an average clearance rate (CL) of about 238 mL/day, an average apparent
volume of distribution (Vss) of about 6 L, and an average serum half-life of
about
22 days is achieved.
[0036] In some embodiments, the number of IV doses at about 14-day
intervals
required to achieve steady state is about 5 to about 8 doses.
[0037] In some embodiments, the administration is subcutaneous (SC)
administration. In certain embodiments, the dosage is 100 mg administered as a
single SC dose, or is administered weekly, bi-weekly, or monthly. In some
embodiments, a T. or Tmax ss of between about 2 and about 10 days is achieved
after SC administration.
[0038] In some embodiments, a single SC administration of about 100 mg
achieves one or more pharmacokinetic characteristics chosen from: a T. of
about 2 to about 10 days, a C. of about 4 to about 21 pg/mL, an area under the
plasma concentration-time curve from time zero to time of last measurable
concentration (AUCiast) of about 175 to about 666 pg day/mL, and an area under
the plasma concentration-time curve from time zero to infinity (AUC.) of about
204 to about 751 pg day/mL.
[0039] In some embodiments, a single SC administration of about 100 mg to a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from: a T. of about 6 days, a C. of about 13 pg/mL, an AUCiast of
about 421 pg day/mL, and an AUC.L. of about 477 pg day/mL.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
[0040] In some embodiments, a single SC administration of about 100 mg
achieves one or more pharmacokinetic characteristics chosen from a clearance
rate (CL/F) of between about 118 and about 432 mL/day, an apparent volume of
distribution (Vz/F) of between about 5 and about 12 L, and a serum half-life
of
about 15 days to about 34 days.
[0041] In some embodiments, a single SC administration of about 100 mg to a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from an average clearance rate (CL/F) of about 275 mL/day, an apparent
volume of distribution (\//F) of about 8 L, and a serum half-life of about 25
days.
[0042] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 7-day (weekly) intervals to achieve a steady state,
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
Tmax ss of about 2 to about 7 days, a Cmax ss of about 37 to about 93 pg/mL,
an
AUCTss of about 248 to about 638 pg day/mL, and a Ctrough ss of about 38 to
about
80 pg/mL is achieved.
[0043] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered to a population of subjects at about 7-day (weekly) intervals
to
achieve a steady state, wherein one or more steady state pharmacokinetic
characteristics chosen from: an average Tmax ss of about 4 days, an average C.
ss of about 65 pg/mL, an average AUCT ss of about 443 pg day/mL, and a Ctrough
ss
of about 59 pg/mL is achieved.
[0044] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 7-day (weekly) intervals to achieve a steady state,
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
clearance rate (CLss/F) of between about 168 and about 396 mL/day, an
apparent volume of distribution (\//F) of between about 7 and about 15 L, and
a
serum half-life of about 22 days to about 35 days is achieved.
[0045] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered to a population of subjects at about 7-day (weekly) intervals
to
achieve a steady state, wherein one or more steady state pharmacokinetic
characteristics chosen from an average clearance rate (CLss) of about 282
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
11
mL/day, an average apparent volume of distribution (Vz/F) of about 11 L, and
an
average serum half-life of about 28 days is achieved.
[0046] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 14-day (bi-weekly) intervals to achieve a steady
state,
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
Tmax ss of about 2 to about 7 days, a Cmax ss of about 30 to about 49 pg/mL,
an
AUCT ss of about 424 to about 567 pg day/mL, and a Ctrough ss of about 21 to
about 40 pg/mL is achieved.
[0047] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered to a population of subjects at about 14-day (bi-weekly)
intervals
to achieve a steady state, wherein one or more steady state pharmacokinetic
characteristics chosen from: an average Tmax ss of about 4 days, an average C.
ss of about 39 pg/mL, an average AUCT ss of about 495 pg day/mL, and a Ctrough
ss
of about 30 pg/mL is achieved.
[0048] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 14-day (bi-weekly) intervals to achieve a steady
state,
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
clearance rate (CLss/F) of between about 172 and about 240 mL/day, an
apparent volume of distribution (Vz/F) of between about 6 and about 10 L, and
a
serum half-life of about 19 days to about 37 days is achieved.
[0049] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered to a population of subjects at about 14-day (bi-weekly)
intervals
to achieve a steady state, wherein one or more steady state pharmacokinetic
characteristics chosen from an average clearance rate (CL) of about 406
mL/day, an average apparent volume of distribution (Vz/F) of about 8 L, and an
average serum half-life of about 28 days is achieved.
[0050] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 30-day (monthly) intervals to achieve a steady
state,
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
Tmax ss of about 3 to about 8 days, a Cmax ss of about 14 to about 34 pg/mL,
an
AUCT ss of about 326 to about 641 pg day/mL, and a Ctrough ss of about 6 to
about
15 pg/mL is achieved.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
12
[0051] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered to a population of subjects at about 30-day (monthly)
intervals
to achieve a steady state, wherein one or more steady state pharmacokinetic
characteristics chosen from: an average Tmax ss of about 6 days, an average C.
ss of about 49 pg/mL, an average AUCT ss of about 483 pg day/mL, and a Ctrough
ss
of about 11 pg/mL is achieved.
[0052] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 30-day (monthly) intervals to achieve a steady
state,
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
clearance rate (CLss/F) of between about 152 and about 302 mL/day, an
apparent volume of distribution (Vz/F) of between about 5 and about 17 L, and
a
serum half-life of about 19 days to about 47 days is achieved.
[0053] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered to a population of subjects at about 30-day (monthly)
intervals
to achieve a steady state, wherein one or more steady state pharmacokinetic
characteristics chosen from an average clearance rate (CL) of about 227
mL/day, an average apparent volume of distribution (Vz/F) of about 11 L, and
an
average serum half-life of about 33 days is achieved.
[0054] In certain embodiments, the administration of a sufficient number of
doses
of an anti-IFN-alpha antibody or antigen-binding fragment thereof suppresses
an
IFN pharmacodynamic signature. In some embodiments, the IFN
pharmacodynamic signature is a Type I IFN-alpha inducible expression profile.
The Type I IFN-alpha inducible expression profile can comprises the up-
regulated
expression of a gene marker set comprising IF144, IF127, IF144L, NAPTP,
LAMP3, LY6E, RSAD2, HERC5, IF16, I5G15, 0A53, RTP4, IFIT1, MX1,
SIGLEC1, 0A52, USP18, OAS1, EPSTI1, PLSCR1 and IFRG28. In some
embodiments, the anti-IFN antibody or antigen-binding fragment thereof
neutralizes the pharmacodynamic expression profile of the patient by at least
10%, at least 20%, at least 30% or at least 40%.
In certain embodiments the method the reduces at least one disease symptom .
In certain embodiments, the reduction in symptoms reduces the SLEDAI or BILAG
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
13
score. In certain embodiments, the SLEDAI score is reduced by at least 1, at
least 2, at
least 3, at least 4, or more points.
BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES
[0055] FIG. 1 is a graph showing mean sifalimumab concentrations over 14 IV
infusion doses. Sifalimumab was administered at 0.3 mg/kg (.), 1.0 mg/kg (=),
3.0 mg/kg (v) and 10 mg/kg (A) doses. Error bars show the standard deviation.
[0056] FIG. 2A is a graph showing frequency of anti-sifalimumab antibody
titer in
a group administered 0.3 mg/kg IV infusion doses.
[0057] FIG. 2B is a graph showing frequency of anti-sifalimumab antibody
titer in
a group administered 1.0 mg/kg IV infusion doses.
[0058] FIG. 2C is a graph showing frequency of anti-sifalimumab antibody
titer in
a group administered 3.0 mg/kg IV infusion doses.
[0059] FIG. 2D is a graph showing frequency of anti-sifalimumab antibody
titer in
a group administered 10 mg/kg IV infusion doses.
[0060] FIG. 3 is a graph showing steady state clearance of sifalimumab by
titer of
anti-sifalimumab antibodies (IM titer). Sifalimumab was administered at 0.3
mg/kg
(.), 1.0 mg/kg (.),3.0 mg/kg (v) and 10 mg/kg (A) IV infusion doses.
[0061] FIG. 4 is a graph showing steady state clearance of sifalimumab by
IM
status of patients receiving 0.3, 1.0, 3.0 and 10 mg/kg doses of sifalimumab.
IM+
and IM- patients are patients testing positive or negative for the presence of
anti-
sifalimumab antibodies, respectively.
[0062] FIG. 5 summarizes the final model goodness-of-fit plots for
sifalimumab
serum concentrations. The thin solid line (diagonal and horizontal) and thick
line
represent line of unity and loess fit, respectively. Panel (a) shows
population
predictions versus observed serum concentrations, panel (b) shows individual
predictions versus observed serum concentrations, panel (c) shows population
predictions versus weighted residuals, and panel (d) shows time versus
weighted
residuals.
[0063] FIG. 6 provides graphs depicting a visual predictive check for
sifalimumab
serum concentrations. Panels shows the time versus serum concentration graphs
corresponding to 0.3, 1.0, 3.0 and 10 mg/kg doses, respectively. Observed
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
14
median (solid line) and corresponding simulation based 95% confidence interval
(light area between shaded areas) and the 5% and 95% data percentiles (dashed
lines) and corresponding simulations based 95% Cl (shaded area) are shown.
[0064] FIG. 7 is a graph showing the similarity of predicted PK profiles
(median,
5th and 95th percentiles) following fixed (200 mg every 14 days) and body
weight
based (3 mg/kg every 14 days) IV dosing of sifalimumab.
[0065] FIG. 8 is a graph showing predicted serum concentrations following
200,
600, and 1200 mg monthly IV dosing of sifalimumab (with single loading dose at
day 14).
[0066] FIG. 9 is a graph showing the mean concentration of sifalimumab over
168
days. Sifalimumab was administered in a single subcutaneous dose (.), weekly
(=), bi-weekly (A) or monthly (=).
[0067] FIG. 10 is a graph showing sifalimumab clearance by immunogenicity
titer
in IM-' and IM- patients. Sifalimumab was administered in a single
subcutaneous
dose (.),weekly (A), bi-weekly (v) or monthly (*).
[0068] FIG. 11 is a graph showing the inhibition of the type 1 IFN
pharmacodynamic gene signature by sifalimumab. Sifalimumab was
administered in a single 100 mg subcutaneous dose once, once a week, bi-
weekly, or monthly.
DETAILED DESCRIPTION
[0069] It must be noted that, as used in this specification and the
appended
claims, the singular forms "a", "an" and "the" include plural referents unless
the
context clearly dictates otherwise. The terms "a" (or "an"), as well as the
terms
"one or more," and "at least one" can be used interchangeably herein.
[0070] Furthermore, "and/or" where used herein is to be taken as specific
disclosure of each of the two specified features or components with or without
the
other. Thus, the term "and/or" as used in a phrase such as "A and/or B" herein
is
intended to include "A and B," "A or B," "A" (alone), and "B" (alone).
Likewise, the
term "and/or" as used in a phrase such as "A, B, and/or C" is intended to
encompass each of the following embodiments: A, B, and C; A, B, or C; A or C;
A
or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
[0071] It is understood that wherever embodiments are described herein with
the
language "comprising," otherwise analogous embodiments described in terms of
"consisting of' and/or "consisting essentially of' are also provided.
[0072] Unless defined otherwise, all technical and scientific terms used
herein
have the same meaning as commonly understood by one of ordinary skill in the
art to which this disclosure is related. For example, the Concise Dictionary
of
Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press;
The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press;
and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised,
2000, Oxford University Press, provide one of skill with a general dictionary
of
many of the terms used in this disclosure.
[0073] Units, prefixes, and symbols are denoted in their Systeme
International de
Unites (SI) accepted form. Numeric ranges are inclusive of the numbers
defining
the range. Unless otherwise indicated, amino acid sequences are written left
to
right in amino to carboxy orientation. The headings provided herein are not
limitations of the various aspects or embodiments of the disclosure, which can
be
had by reference to the specification as a whole. Accordingly, the terms
defined
immediately below are more fully defined by reference to the specification in
its
entirety. Amino acids are referred to herein by either their commonly known
three
letter symbols or by the one-letter symbols recommended by the IUPAC-IUB
Biochemical Nomenclature Commission. Nucleotides, likewise, are referred to by
their commonly accepted single-letter codes.
[0074] As used herein, the term "autoimmune disease" refers to a disorder,
disease state or condition associated with the formation of autoantibodies
reactive with the patient's own cells to form antigen-antibody complexes. The
term "autoimmune disease" includes conditions such as, e.g., systemic lupus
erythematosus, as well as those disorders which are triggered by a specific
external agent, e.g., acute rheumatic fever. Examples of autoimmune disorders
include, but are not limited to, autoimmune hemolytic anemia, autoimmune
hepatitis, Berger's disease, chronic fatigue syndrome, Crohn's disease,
dermatomyositis, fibromyalgia, Graves' disease, Hashimoto's thyroiditis,
idiopathic thrombocytopenia purpura, lichen planus, multiple sclerosis,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
16
myasthenia gravis, psoriasis, rheumatic fever, rheumatoid arthritis,
scleroderma,
Sjogren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative
colitis, and vitiligo. In specific embodiments, the autoimmune disease is
systemic
lupus erythematosus, scleroderma, or myositis.
[0075] As used herein, the term "antibody" is used in its broadest sense
and
includes monoclonal antibodies, polyclonal antibodies, multivalent antibodies,
multispecific antibodies, chimeric antibodies, and humanized antibodies. The
term "antibody" as referred to herein includes whole antibodies. An "antibody"
refers to a glycoprotein comprising at least two heavy (H) chains and two
light (L)
chains inter-connected by disulfide bonds, or an antigen binding portion
thereof.
Each heavy chain is comprised of a heavy chain variable region (abbreviated
herein as VH) and a heavy chain constant region. The heavy chain constant
region is comprised of three domains, CHI, CH2 and CH3. Each light chain is
comprised of a light chain variable region (abbreviated herein as VL) and a
light
chain constant region. The light chain constant region is comprised of one
domain, CL. The VH and VL regions can be further subdivided into regions of
hypervariability, termed complementarity determining regions (CDR),
interspersed with regions that are more conserved, termed framework regions
(FR). Each VH and VL is composed of three CDRs and four FRs, arranged from
amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2,
CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains
contain a binding domain that interacts with an antigen. The constant regions
of
the antibodies can mediate the binding of the immunoglobulin to host tissues
or
factors, including various cells of the immune system (e.g., effector cells)
and the
first component (C1q) of the classical complement system.
[0076] The terms "fragment thereof," "antigen-binding fragment" and
"antigen-
binding portion" of an antibody (or simply "antibody portion") are used
interchangeably herein, and refer to one or more fragments of an antibody that
retain the ability to specifically bind to an antigen (e.g., IFN-alpha). It
has been
shown that the antigen-binding function of an antibody can be performed by
fragments of a full-length antibody. Examples of binding fragments encompassed
within the term "antigen-binding portion" of an antibody include (i) a Fab
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
17
fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains;
(ii) a F(ab1)2 fragment, a bivalent fragment comprising two Fab fragments
linked
by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of
the VH
and CHI domains; (iv) a Fv fragment consisting of the VL and VH domains of a
single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature
341:544-546), which consists of a VH domain; and (vi) an isolated
complementarity determining region (CDR). Furthermore, although the two
domains of the Fv fragment, VL and VH, are coded for by separate genes, they
can be joined, using recombinant methods, by a synthetic linker that enables
them to be made as a single protein chain in which the VL and VH regions pair
to
form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et
al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad.
Sci.
USA 85:5879-5883). Such single chain antibodies are also intended to be
encompassed within the term "antigen-binding portion" of an antibody. These
antibody fragments are obtained using conventional techniques known to those
with skill in the art, and the fragments are screened for utility in the same
manner
as are intact antibodies.
[0077] An "isolated antibody", as used herein, is intended to refer to an
antibody
that is substantially free of other antibodies having different antigenic
specificities
(e.g., an isolated antibody that specifically binds IFN-alpha is substantially
free of
antibodies that specifically bind antigens other than IFN-alpha). An isolated
antibody that specifically binds IFN-alpha can, however, have cross-reactivity
to
other antigens, such as IFN-alpha molecules from other species. Moreover, an
isolated antibody can be substantially free of other cellular material and/or
chemicals.
[0078] The terms "monoclonal antibody" or "monoclonal antibody composition"
as
used herein refer to a preparation of antibody molecules of single molecular
composition. A monoclonal antibody composition displays a single binding
specificity and affinity for a particular epitope.
[0079] The term "human antibody", as used herein, is intended to include
antibodies having variable regions in which both the framework and CDR regions
are derived from human germline immunoglobulin sequences. Furthermore, if the
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
18
antibody contains a constant region, the constant region also is derived from
human germline immunoglobulin sequences. The human antibodies of the
disclosure can include amino acid residues not encoded by human germline
immunoglobulin sequences (e.g., mutations introduced by random or site-
specific
mutagenesis in vitro or by somatic mutation in vivo). However, the term "human
antibody", as used herein, is not intended to include antibodies in which CDR
sequences derived from the germline of another mammalian species, such as a
mouse, have been grafted onto human framework sequences.
[0080] The term "human monoclonal antibody" refers to antibodies displaying
a
single binding specificity which have variable regions in which both the
framework
and CDR regions are derived from human germline immunoglobulin sequences.
In one embodiment, the human monoclonal antibodies are produced by a
hybridoma which includes a B cell obtained from a transgenic nonhuman animal,
e.g., a transgenic mouse, having a genome comprising a human heavy chain
transgene and a light chain transgene fused to an immortalized cell.
[0081] The term "recombinant human antibody", as used herein, includes all
human antibodies that are prepared, expressed, created or isolated by
recombinant means, such as (a) antibodies isolated from an animal (e.g., a
mouse) that is transgenic or transchromosomal for human immunoglobulin genes
or a hybridoma prepared therefrom (described further below), (b) antibodies
isolated from a host cell transformed to express the human antibody, e.g.,
from a
transfectoma, (c) antibodies isolated from a recombinant, combinatorial human
antibody library, and (d) antibodies prepared, expressed, created or isolated
by
any other means that involve splicing of human immunoglobulin gene sequences
to other DNA sequences. Such recombinant human antibodies have variable
regions in which the framework and CDR regions are derived from human
germline immunoglobulin sequences. In certain embodiments, however, such
recombinant human antibodies can be subjected to in vitro mutagenesis (or,
when an animal transgenic for human Ig sequences is used, in vivo somatic
mutagenesis) and thus the amino acid sequences of the VH and VL regions of
the recombinant antibodies are sequences that, while derived from and related
to
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
19
human germline VH and VL sequences, may not naturally exist within the human
antibody germline repertoire in vivo.
[0082] As used herein, "isotype" refers to the antibody class (e.g., IgM or
IgG1)
that is encoded by the heavy chain constant region genes.
[0083] The antibodies herein specifically include "chimeric" antibodies in
which a
portion of the heavy and/or light chain is identical with or homologous to
corresponding sequences in antibodies derived from a particular species or
belonging to a particular antibody class or subclass, while the remainder of
the
chain(s) is identical with or homologous to corresponding sequences in
antibodies derived from another species or belonging to another antibody class
or
subclass, as well as fragments of such antibodies, so long as they exhibit the
desired biological activity (U.S. Patent No. 4,816,567; and Morrison et al,
Proc.
Natl. Acad. Sci. USA 8/:6851-6855 (1984)).
[0084] Basic antibody structures in vertebrate systems are relatively well
understood. See, e.g., Harlow et al. (1988) Antibodies: A Laboratory Manual
(2nd ed.; Cold Spring Harbor Laboratory Press).
[0085] In the case where there are two or more definitions of a term that
is used
and/or accepted within the art, the definition of the term as used herein is
intended to include all such meanings unless explicitly stated to the
contrary. A
specific example is the use of the term "complementarity determining region"
("CDR") to describe the non-contiguous antigen combining sites found within
the
variable region of both heavy and light chain polypeptides. This particular
region
has been described by Kabat et al. (1983) U.S. Dept. of Health and Human
Services, "Sequences of Proteins of Immunological Interest" and by Chothia and
Lesk, J. Mol. Biol. /96:901-917 (1987), which are incorporated herein by
reference, where the definitions include overlapping or subsets of amino acid
residues when compared against each other. Nevertheless, application of either
definition to refer to a CDR of an antibody or variants thereof is intended to
be
within the scope of the term as defined and used herein. The appropriate amino
acid residues that encompass the CDRs as defined by each of the above cited
references are set forth below in TABLE 1 as a comparison. The exact residue
numbers that encompass a particular CDR will vary depending on the sequence
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
and size of the CDR. Those skilled in the art can routinely determine which
residues comprise a particular CDR given the variable region amino acid
sequence of the antibody.
TABLE 1
CDR Definition&
Kabat Chothia
VH CDR1 31-35 26-32
VH CDR2 50-65 52-58
VH CDR3 95-102 95-102
VL CDR1 24-34 26-32
VL CDR2 50-56 50-52
VL CDR3 89-97 91-96
'Numbering of all CDR definitions in Table 1 is according to the
numbering conventions set forth by Kabat et al. (see below).
[0086] Kabat et al. also defined a numbering system for variable domain
sequences that is applicable to any antibody. One of ordinary skill in the art
can
unambiguously assign this system of "Kabat numbering" to any variable domain
sequence, without reliance on any experimental data beyond the sequence
itself.
As used herein, "Kabat numbering" refers to the numbering system set forth by
Kabat et al. (1983) U.S. Dept. of Health and Human Services, "Sequence of
Proteins of Immunological Interest." Unless otherwise specified, references to
the
numbering of specific amino acid residue positions in an anti-IL-33 antibody
or
antigen-binding fragment, variant, or derivative thereof of the present
disclosure
are according to the Kabat numbering system.
[0087] Antibodies or antigen-binding fragments, variants, or derivatives
thereof of
the disclosure include, but are not limited to, polyclonal, monoclonal,
multispecific, mouse, human, humanized, primatized, or chimeric antibodies,
single-chain antibodies, epitope-binding fragments, e.g., Fab, Fab' and
F(abl)2,
Fd, Fvs, single-chain Fvs (scFv), disulfide-linked Fvs (sdFv), fragments
comprising either a VL or VH domain, fragments produced by a Fab expression
library, and anti-idiotypic (anti-ld) antibodies (including, e.g., anti-ld
antibodies to
anti-IL-33 antibodies disclosed herein). ScFv molecules are known in the art
and
are described, e.g., in U.S. Pat. No. 5,892,019.
[0088] As used herein, the term "affinity" refers to a measure of the
strength of
the binding of an individual epitope with the CDR of an immunoglobulin
molecule.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
21
See, e.g., Harlow et al. (1988) Antibodies: A Laboratory Manual (Cold Spring
Harbor Laboratory Press, 2nd ed.) pages 27-28.
[0089] As used herein, "specific binding" refers to antibody binding to a
predetermined antigen. Typically, the antibody binds with a dissociation
constant
(KD) of 10-8 M or less, and binds to the predetermined antigen with a KD that
is at
least two-fold less than its KD for binding to a non-specific antigen (e.g.,
BSA,
casein) other than the predetermined antigen or a closely-related antigen. The
phrases "an antibody recognizing an antigen" and "an antibody specific for an
antigen" are used interchangeably herein with the term "an antibody which
binds
specifically to an antigen".
[0090] The term uKassocu or "Ka", as used herein, is intended to refer to
the
association rate of a particular antibody-antigen interaction, whereas the
term
"Kd,s" or "Kd," as used herein, is intended to refer to the dissociation rate
of a
particular antibody-antigen interaction. The term " KD", as used herein, is
intended to refer to the dissociation constant, which is obtained from the
ratio of
Kd to Ka and is expressed as a molar concentration (M). KD values for
antibodies
can be determined using methods well established in the art. One method for
determining the KD of an antibody is by using surface plasmon resonance, e.g.,
by using a biosensor system such as a Biacore® system.
[0091] As used herein, the term "high affinity" for an IgG antibody refers
to an
antibody having a KD of 10-8 M or less, 10-9 M or less, or 10-19 M or less.
However, "high affinity" binding can vary for other antibody isotypes. For
example, "high affinity" binding for an IgM isotype refers to an antibody
having a
KD of 10-7 M or less, or 10-8 M or less.
[0092] Anti-IFN-alpha binding molecules, e.g., antibodies or antigen-
binding
fragments, variants or derivatives thereof of the disclosure can also be
described
or specified in terms of their binding affinity to a polypeptide of the
disclosure,
e.g., IFN- alpha, e.g., human, primate, murine, or any combination of human,
primate and murine IFN-alpha. Exemplary binding affinities include those with
a
dissociation constant or KD less than 5 x 10-2 M, 10-2 M, 5 x 10-3 M, 10-3 M,
5 x 10-
4 M, 10-4 M, 5 x 10-5 M, 10-5 M, 5 x 10-6 M, 10-6 M, 5 x 10-7 M, 10-7 M, 5 x
10-8 M,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
22
10-8 M, 5 x 10-9 M, 10-9 M, 5 x 10-19 M, 10-19 M, 5 x 10-11 M, 10-11 M, 5 x 10-
12 M,
10-12 M, 5 x 10-13 M, 10-13 M, 5 x 10-14 M, 10-14 M, 5 x 10-15 M, or 10-15 M.
[0093] As used herein, the terms "treat" or "treatment" refer to both
therapeutic
treatment and prophylactic or preventative measures, wherein the object is to
prevent or slow down (lessen) an undesired physiological change or disorder,
such as the progression of an inflammatory condition. Beneficial or desired
clinical results include, but are not limited to, alleviation of symptoms,
diminishment of extent of disease, stabilized (i.e., not worsening) state of
disease, delay or slowing of disease progression, amelioration or palliation
of the
disease state, and remission (whether partial or total), whether detectable or
undetectable. "Treatment" can also mean prolonging survival as compared to
expected survival if not receiving treatment. Those in need of treatment
include
those already with the condition or disorder as well as those prone to have
the
condition or disorder or those in which the condition or disorder is to be
prevented.
[0094] The terms "effective amount" or "amount effective to" or
"therapeutically
effective amount" includes reference to a dosage of a therapeutic agent
sufficient
to produce a desired result.
[0095] By "subject" or "individual" or "animal" or "patient" or "mammal,"
is meant
any subject, particularly a mammalian subject, for whom diagnosis, prognosis,
or
therapy is desired. As used herein, the term "subject" includes any human or
nonhuman animal. The term "nonhuman animal" includes all vertebrates, e.g.,
mammals and non-mammals, such as nonhuman primates, sheep, dogs, cats,
horses, cows, bears, chickens, amphibians, reptiles, etc. As used herein,
phrases
such as "a subject that would benefit from administration of an anti-IFN-
alpha antibody" includes subjects, such as mammalian subjects, that would
benefit from administration of an anti-IFN-alpha antibody used, e.g., for
detection
of an anti-IFN-alpha polypeptide (e.g., for a diagnostic procedure) and/or
from
treatment, i.e., palliation or prevention of a disease, with an anti-IFN-alpha
antibody.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
23
Interferon Alpha
[0096] The terms "interferon alpha" and "IFN-alpha" are used
interchangeably
and intended to refer to IFN-alpha proteins encoded by a functional gene of
the
interferon alpha gene locus with 75% or greater sequence identity to IFN-alpha
1
(Genbank number NP-076918 or protein encoded by Genbank number NM-
024013). Examples of IFN-alpha subtypes include IFN-alpha 1, alpha 2a, alpha
2b, alpha 4, alpha 5, alpha 6, alpha 7, alpha 8, alpha 10, alpha 13, alpha 14,
alpha 16, alpha 17 and alpha 21. The term "interferon alpha" is intended to
encompass recombinant forms of the various IFN-alpha subtypes, as well as
naturally occurring preparations that comprise IFN-alpha proteins, such as
leukocyte IFN and lymphoblastoid IFN.
Interferon Alpha Antibodies
[0097] The present disclosure is directed to methods of treating an
autoimmune
disorder in a subject in need of such treatment comprising administering to
the
subject an anti-IFN-alpha antibody. Anti-IFN-alpha antibodies can be found in,
for
example, U.S. Pat. No. 7,741,449, and can further include chimeric, humanized,
or human versions of these antibodies (if not already a chimeric, humanized,
or
human version), and can further include fragments or derivatives thereof.
SLE
[0098] Patients suffering from SLE may exhibit any of a number of symptoms
as
discussed in, e.g., International Application No. PCT/US2007/024941, or may
have a clinical SLEDAI score or BILAG score as discussed in the same. These
symptoms may include fatigue, organ damage, malar rash, and alopecia. The
patient may be scored using a known clinical scoring system, e.g., SLEDAI
which
is an index of SLE disease activity as measured and evaluated within the last
10
days (Bombardier C, Gladman D D, Urowitz M B, Caron D, Chang C H and the
Committee on Prognosis Studies in SLE: Derivation of the SLEDAI for Lupus
Patients. Arthritis Rheum 35:630-640, 1992.). Disease activity under the
SLEDAI
scoring system can range from 0 to 105. The following categories of SLEDAI
activity have been defined: no activity (SLEDAI = 0); mild activity (SLEDAI =
1-
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
24
5); moderate activity (SLEDAI = 6-10); high activity (SLEDAI = 11-19); very
high
activity (SLEDAI = 20 or higher). (Griffiths, et al., Assessment of Patients
with
Systemic Lupus Erythematosus and the use of Lupus Disease Activity Indices).
[0099] Another disease scoring index is the BILAG index which is an
activity
index of SLE that is based on specific clinical manifestations in eight organ
systems: general, mucocutaneous, neurological, musculoskeletal,
cardiovascular,
respiratory, renal, and hematology results. Scoring is based on a letter
system,
but weighted numerical scores can also be assigned to each letter, making it
possible to calculate a BILAG score in the range of 0-72. (Griffiths, et al.,
Assessment of Patients with Systemic Lupus Erythematosus and the use of
Lupus Disease Activity Indices). Other scoring indices include the PGA score,
the composite responder index (CRI), and the ANAM4Tm test. The methods
described herein, e.g., of treating an autoimmune disorder, may be used for
any
subject identified as having any activity level of disease activity as
measured by
any classification methodology known in the art, e.g., mild, moderate, high,
or
very high. The methods described herein, e.g., of treating an autoimmune
disorder, may result in a decrease in a patient's symptoms or may result in an
improvement in a score of disease for the patient's type I IFN or an IFN-alpha-
inducible disease, disorder, or condition.
Monoclonal Antibodies 13H5, 13H7 and 7H9
[0100] In certain embodiments, antibodies for use in the treatment methods
of the
disclosure include the human monoclonal antibodies 13H5, 13H7, and 7H9,
isolated and structurally characterized as described in the U.S. Patent No.
7,741,449. The VH amino acid sequences of 13H5, 13H7, and 7H9 are shown in
SEQ ID NOs: 19, 20, and 21, respectively. The VL amino acid sequences of
13H5, 13H7, and 7H9 are shown in SEQ ID NOs: 22, 23 and 24, respectively.
Given that each of these antibodies can bind to IFN-alpha, the VH and VL
sequences can be "mixed and matched" to create other anti-IFN-alpha binding
molecules of the disclosure. IFN-alpha binding or neutralizing activity of
such
"mixed and matched" antibodies can be tested using the binding assays
described above and in the Examples (e.g., ELISA, Biacore analysis, Daudi cell
proliferation assay). In certain embodiments, the VH sequences of 13H5 and 7H9
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
are mixed and matched, since these antibodies use VH sequences derived from
the same germline sequence (VH 1-18) and thus they exhibit structural
similarity.
Additionally or alternatively, the VL sequences of 13H5, 13H7 and 7H9 can be
mixed and matched, since these antibodies use VL sequences derived from the
same germline sequence (Vk A27) and thus they exhibit structural similarity.
[0101] Accordingly, in one aspect, an antibody for use in the methods of
the
disclosure is an isolated monoclonal antibody, or antigen binding portion
thereof,
comprising:
(a) a heavy chain variable region comprising an amino acid sequence chosen
from SEQ ID NOs: 19, 20, and 21; and
(b) a light chain variable region comprising an amino acid sequence chosen
from
SEQ ID NOs: 22, 23, and 24;
wherein the antibody inhibits the biological activity of interferon alpha.
[0102] In certain embodiments, heavy and light chain combinations include:
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID
NO: 19; and
(b) a light chain variable region comprising the amino acid sequence of SEQ ID
NO:22; or
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID
NO: 20; and
(b) a light chain variable region comprising the amino acid sequence of SEQ ID
NO:23; or
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID
NO: 21; and
(b) a light chain variable region comprising the amino acid sequence of SEQ ID
NO:24.
[0103] In another aspect, the treatment methods of the present disclosure
comprise the administration of antibodies that comprise the heavy chain and
light
chain CDR1s, CDR2s, and CDR3s of 13H5, 13H7, and 7H9, or combinations
thereof. The amino acid sequences of the VH CDR1s of 13H5, 13H7, and 7H9
are shown in SEQ ID NOs: 1, 2, and 3. The amino acid sequences of the VH
CDR2s of 13H5, 13H7, and 7H9 are shown in SEQ IN NOs: 4, 5, and 6. The
amino acid sequences of the VH CDR3s of 13H5, 13H7, and 7H9 are shown in
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
26
SEQ IN NOs: 7, 8, and 9. The amino acid sequences of the VL CDR1s of 13H5,
13H7, and 7H9 are shown in SEQ IN NOs: 10, 11, and 12. The amino acid
sequences of the VL CDR2s of 13H5, 13H7, and 7H9 are shown in SEQ IN NOs:
13, 14, and 15. The amino acid sequences of the VL CDR3s of 13H5, 13H7, and
7H9 are shown in SEQ IN NOs: 16, 17, and 18. The CDR regions are delineated
using the Kabat system (Kabat. E. A., et al. (1991) Sequences of Proteins of
Immunological Interest, Fifth Edition, U.S. Department of Health and Human
Services, NIH Publication No. 91-3242).
[0104] Given that each of these antibodies was selected based on IFN
binding
activity and that antigen-binding specificity is provided primarily by the
CDR1, 2
and 3 regions, the VH CDR1, 2 and 3 sequences and VL CDR1, 2 and 3
sequences can be "mixed and matched" (i.e., CDRs from different antibodies can
be mixed and match, although each antibody must contain a VH CDR1, 2 and 3
and a VL CDR1, 2 and 3) to create other anti-IFN-alpha molecules of the
disclosure. IFN-alpha binding of such "mixed and matched" antibodies can be
tested using the binding assays described in the Examples (e.g., ELISA and/or
Biacore). In certain embodiments, when VH CDR sequences are mixed and
matched, the CDR1, CDR2 and/or CDR3 sequence from a particular VH
sequence is replaced with a structurally similar CDR sequence(s). Likewise,
when VL CDR sequences are mixed and matched, the CDR1, CDR2 and/or
CDR3 sequence from a particular VL sequence can be replaced with a
structurally similar CDR sequence(s). For example, the VH CDR1s of 13H5 and
7H9 share some structural similarity and therefore are amenable to mixing and
matching. It will be readily apparent to the ordinarily skilled artisan that
novel VH
and VL sequences can be created by substituting one or more VH and/or VL
CDR region sequences with structurally similar sequences from the CDR
sequences disclosed herein for monoclonal antibodies 13H5, 13H7 and 7H9.
[0105] Accordingly, in another aspect, the methods of the present
disclosure
provide for administration of an isolated monoclonal antibody, or antigen
binding
portion thereof comprising:
(a) a heavy chain variable region CDR1 comprising an amino acid sequence
chosen from SEQ ID NOs: 1, 2, and 3;
CA 02836926 2013-11-20
WO 2012/162367
PCT/US2012/039098
27
(b) a heavy chain variable region CDR2 comprising an amino acid sequence
chosen from SEQ ID NOs: 4, 5, and 6;
(c) a heavy chain variable region CDR3 comprising an amino acid sequence
chosen from SEQ ID NOs: 7, 8, and 9;
(d) a light chain variable region CDR1 comprising an amino acid sequence
chosen from SEQ ID NOs: 10, 11, and 12;
(e) a light chain variable region CDR2 comprising an amino acid sequence
chosen from SEQ ID NOs: 13, 14, and 15; and
(f) a light chain variable region CDR3 comprising an amino acid sequence
chosen from SEQ ID NOs: 16, 17, and 18;
wherein the antibody the antibody inhibits the biological activity of
interferon
alpha.
[0106] In one embodiment, the antibody comprises:
(a) a heavy chain variable region CDR1 comprising SEQ ID NO: 1;
(b) a heavy chain variable region CDR2 comprising SEQ ID NO: 4;
(c) a heavy chain variable region CDR3 comprising SEQ ID NO: 7;
(d) a light chain variable region CDR1 comprising SEQ ID NO: 10;
(e) a light chain variable region CDR2 comprising SEQ ID NO: 13; and
(f) a light chain variable region CDR3 comprising SEQ ID NO: 16.
[0107] In another embodiment, the antibody comprises:
(a) a heavy chain variable region CDR1 comprising SEQ ID NO: 2;
(b) a heavy chain variable region CDR2 comprising SEQ ID NO: 5;
(c) a heavy chain variable region CDR3 comprising SEQ ID NO: 8;
(d) a light chain variable region CDR1 comprising SEQ ID NO: 11;
(e) a light chain variable region CDR2 comprising SEQ ID NO: 14; and
(f) a light chain variable region CDR3 comprising SEQ ID NO: 17.
[0108] In another embodiment, the antibody comprises:
(a) a heavy chain variable region CDR1 comprising SEQ ID NO: 3;
(b) a heavy chain variable region CDR2 comprising SEQ ID NO: 6;
(c) a heavy chain variable region CDR3 comprising SEQ ID NO: 9;
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
28
(d) a light chain variable region CDR1 comprising SEQ ID NO: 12;
(e) a light chain variable region CDR2 comprising SEQ ID NO: 15; and
(f) a light chain variable region CDR3 comprising SEQ ID NO: 18.
Antibodies Having Particular Germline Sequences
[0109] In certain embodiments, an antibody to be administered according to
the
methods of the disclosure comprises a heavy chain variable region from a
particular germline heavy chain immunoglobulin gene and/or a light chain
variable region from a particular germline light chain immunoglobulin gene.
[0110] For example, in one embodiment, the disclosure provides a method of
treating an autoimmune disorder in a subject in need thereof, comprising
administering to the subject an isolated monoclonal antibody, or an antigen-
binding fragment thereof, where the antibody:
(a) comprises a heavy chain variable region of a human VH 1-18 or 4-61 gene;
(b) comprises a light chain variable region of a human VK A27 gene; and
(c) the antibody inhibits the biological activity of interferon alpha.
[0111] In one embodiment, the antibody comprises a heavy chain variable
region
of a human VH 1-18 gene. Examples of antibodies having a VH and VK gene
sequence of VH 1-18 and VK A27, respectively, include 13H5 and 7H9. In
another embodiment, the antibody comprises a heavy chain variable region of a
human VH 4-61 gene. An example of an antibody having a VH and VK gene
sequence of VH 4-61 and VK A27, respectively, is 13H7.
[0112] As used herein, a human antibody comprises heavy or light chain
variable
regions "of' (i.e., the products of) or "derived from" a particular germline
sequence if the variable regions of the antibody are obtained from a system
that
uses human germline immunoglobulin genes. Such systems include immunizing
a transgenic mouse carrying human immunoglobulin genes with the antigen of
interest or screening a human immunoglobulin gene library displayed on phage
with the antigen of interest. A human antibody that is "of' (i.e., the product
of) or
"derived from" a human germline immunoglobulin sequence can be identified as
such by comparing the amino acid sequence of the human antibody to the amino
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
29
acid sequences of human germline immunoglobulins and selecting the human
germline immunoglobulin sequence that is closest in sequence (i.e., greatest %
identity) to the sequence of the human antibody. A human antibody that is "of'
(i.e., the product of) or "derived from" a particular human germline
immunoglobulin sequence can contain amino acid differences as compared to
the germline sequence, due to, for example, naturally-occurring somatic
mutations or intentional introduction of site-directed mutation. However, a
selected human antibody typically is at least 90% identical in amino acids
sequence to an amino acid sequence encoded by a human germline
immunoglobulin gene and contains amino acid residues that identify the human
antibody as being human when compared to the germline immunoglobulin amino
acid sequences of other species (e.g., murine germline sequences). In certain
cases, a human antibody can be at least 95%, or even at least 96%, 97%, 98%,
or 99% identical in amino acid sequence to the amino acid sequence encoded by
the germline immunoglobulin gene. Typically, a human antibody derived from a
particular human germline sequence will display no more than 10 amino acid
differences from the amino acid sequence encoded by the human germline
immunoglobulin gene. In certain cases, the human antibody can display no more
than 5, or even no more than 4, 3, 2, or 1 amino acid difference from the
amino
acid sequence encoded by the germline immunoglobulin gene.
[0113] In yet another embodiment, an antibody for use in the treatment
methods
of the disclosure comprises heavy and light chain variable regions comprising
amino acid sequences that share amino acid similarity with or are homologous
to
the amino acid sequences of the antibodies described herein, where the
antibodies retain the desired functional properties of the anti-IFN-alpha
antibodies
of the disclosure.
[0114] For example, the disclosure provides a method of treating an
autoimmune
disorder in a subject in need thereof, comprising administering to the subject
an
isolated monoclonal antibody, or antigen binding fragment thereof, comprising
a
heavy chain variable region and a light chain variable region, wherein:
(a) the heavy chain variable region comprises an amino acid sequence that is
at
least 80% homologous to an amino acid sequence chosen from SEQ ID NOs:
19, 20, and 21;
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
(b) the light chain variable region comprises an amino acid sequence that is
at
least 80% homologous to an amino acid sequence chosen from SEQ ID NOs:
22, 23, and 24;
(c) the antibody inhibits the biological activity of multiple IFN-alpha
subtypes but
does not substantially inhibit the biological activity of IFN-alpha 21; and
(d) the antibody exhibits at least one of the following properties: (i) the
antibody
does directly inhibit the biological activity of IFN-beta or IFN-omega, but
may
indirectly inhibit the biological activity of IFN-beta or IFN-omega by
interfering
with interferon receptor function; (ii) the antibody inhibits IFN-induced
surface
expression of CD38 or MHC Class I on peripheral blood mononuclear cells;
(iii) the antibody inhibits IFN-induced expression of IP-10 by peripheral
blood
mononuclear cells; (iv) the antibody inhibits dendritic cell development
mediated by systemic lupus erythematosus (SLE) plasma.
[0115] In other embodiments, the VH and/or VL amino acid sequences can be
85%, 90%, 95%, 96%, 97%, 98% or 99% homologous to the sequences set forth
above. An antibody having VH and VL regions having high (i.e., 80% or greater)
homology to the VH and VL regions of SEQ ID NOs: 19, 20, and 21 and 22, 23,
and 24, respectively, can be obtained by mutagenesis (e.g., site-directed or
PCR-
mediated mutagenesis) of nucleic acid molecules encoding SEQ ID NOs: 19, 20,
and 21 and/or 22, 23, and 24, followed by testing of the encoded altered
antibody
for retained function (i.e., the functions set forth in (c) and (d) above)
using the
functional assays described herein.
[0116] As used herein, the percent homology or percent similarity between
two
amino acid sequences is equivalent to the percent identity between the two
sequences. The percent identity between the two sequences is a function of the
number of identical positions shared by the sequences (i.e., % homology =
number of identical positions/total number of positions×100), taking
into
account the number of gaps, and the length of each gap, which need to be
introduced for optimal alignment of the two sequences. The comparison of
sequences and determination of percent identity between two sequences can be
accomplished using a mathematical algorithm, as described in the non-limiting
examples below.
[0117] The percent identity between two amino acid sequences can be
determined using the algorithm of E. Meyers and W. Miller (Comput. Appl.
Biosci., 4:11-17 (1988)) which has been incorporated into the ALIGN program
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
31
(version 2.0), using a PAM120 weight residue table, a gap length penalty of 12
and a gap penalty of 4. In addition, the percent identity between two amino
acid
sequences can be determined using the Needleman and Wunsch (J. Mol. Biol.
48:444-453 (1970)) algorithm which has been incorporated into the GAP program
in the GCG software package (available at http://www.gcg.com), using either a
BLOSSUM62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8,
6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.
[0118] Additionally or alternatively, the protein sequences of the present
disclosure can further be used as a "query sequence" to perform a search
against
public databases to, for example, identify related sequences. Such searches
can
be performed using the XBLAST program (version 2.0) of Altschul, et al. (1990)
J. Mol. Biol. 215:403-10. BLAST protein searches can be performed with the
XBLAST program, score=50, wordlength=3 to obtain amino acid sequences
homologous to the antibody molecules of the disclosure. To obtain gapped
alignments for comparison purposes, Gapped BLAST can be utilized as
described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402. When
utilizing BLAST and Gapped BLAST programs, the default parameters of the
respective programs (e.g., XBLAST and NBLAST) can be used. See
http://www.ncbi.nlm.nih.gov.
Antibodies with Conservative Modifications
[0119] In certain embodiments, an antibody for use in the methods of the
present
disclosure comprises a heavy chain variable region comprising CDR1, CDR2 and
CDR3 sequences and a light chain variable region comprising CDR1, CDR2 and
CDR3 sequences, wherein one or more of these CDR sequences comprise
specified amino acid sequences based on antibodies described herein (e.g.,
13H5, 13H7, or 7H9), or conservative modifications thereof, and wherein the
antibodies retain the desired functional properties of the anti-IFN-alpha
antibodies
of the disclosure. For example, certain antibodies of the disclosure include
those
in which the heavy chain variable region CDR3 sequence comprises the amino
acid sequence of SEQ ID NO: 3, or conservative modifications thereof, and the
light chain variable region CDR3 sequence comprises the amino acid sequence
of SEQ ID NO: 6, or conservative modifications thereof. Accordingly, the
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
32
disclosure provides a method for treating an autoimmune disorder in a subject,
comprising administering to the subject an isolated monoclonal antibody, or
antigen binding fragment thereof, comprising a heavy chain variable region
comprising CDR1, CDR2, and CDR3 sequences and a light chain variable region
comprising CDR1, CDR2, and CDR3 sequences, wherein:
(a) the heavy chain variable region CDR3 sequence comprises the amino acid
sequence chosen from SEQ ID NO: 7, 8, and 9, and conservative
modifications thereof;
(b) the light chain variable region CDR3 sequence comprises the amino acid
sequence chosen from SEQ ID NO: 16, 17, and 18, and conservative
modifications thereof;
(c) the antibody inhibits the biological activity of multiple IFN-alpha
subtypes but
does not substantially inhibit the biological activity of IFN-alpha 21;
(d) the antibody exhibits at least one of the following properties: (i) the
antibody
does not directly inhibit the biological activity of IFN-beta or IFN-omega,
but
may indirectly inhibit the biological activity of IFN-beta or IFN-omega by
interfering with interferon receptor function; (ii) the antibody inhibits IFN-
induced surface expression of CD38 or MHC Class I on peripheral blood
mononuclear cells; (iii) the antibody inhibits IFN-induced expression of IP-10
by peripheral blood mononuclear cells; (iv) the antibody inhibits dendritic
cell
development mediated by systemic lupus erythematosus (SLE) plasma.
[0120] In a further embodiment, the heavy chain variable region CDR2
sequence
comprises the amino acid sequence chosen from amino acid sequences of SEQ
ID NO: 4, 5, and 6, and conservative modifications thereof; and the light
chain
variable region CDR2 sequence comprises the amino acid sequence chosen
from amino acid sequences SEQ ID NO: 13, 14, and 15, and conservative
modifications thereof. In a still further embodiment, the heavy chain variable
region CDR1 sequence comprises the amino acid sequence chosen from amino
acid sequences of SEQ ID NO: 1, 2, and 3, and conservative modifications
thereof; and the light chain variable region CDR1 sequence comprises the amino
acid sequence chosen from amino acid sequences of SEQ ID NO: 10, 11, and
12, and conservative modifications thereof.
[0121] As used herein, the term "conservative sequence modifications" is
intended to refer to amino acid modifications that do not significantly affect
or
alter the binding characteristics of the antibody containing the amino acid
sequence. Such conservative modifications include amino acid substitutions,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
33
additions and deletions. Modifications can be introduced into an antibody of
the
disclosure by standard techniques known in the art, such as site-directed
mutagenesis and PCR-mediated mutagenesis. Conservative amino acid
substitutions are ones in which the amino acid residue is replaced with an
amino
acid residue having a similar side chain. Families of amino acid residues
having
similar side chains have been defined in the art. These families include amino
acids with basic side chains (e.g., lysine, arginine, histidine), acidic side
chains
(e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g.,
glycine,
asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan),
nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline,
phenylalanine, methionine), beta-branched side chains (e.g., threonine,
valine,
isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine,
tryptophan,
histidine). Thus, one or more amino acid residues within the CDR regions of an
antibody of the disclosure can be replaced with other amino acid residues from
the same side chain family and the altered antibody can be tested for retained
function (i.e., the functions set forth in (c) and (d) above) using the
functional
assays described herein.
Antibodies that Bind to the Same Epitope as Anti-IFN-alpha Antibodies of
the Disclosure
[0122] In another embodiment, the disclosure provides a method of treating
an
autoimmune disorder in a subject, comprising administering to the subject an
antibody that binds to the same epitope as do the various human IFN-alpha
antibodies as described herein, such as other human antibodies that bind to
the
same epitope as the 13H5, 13H7, and 7H9 antibodies. The term "epitope" as
used herein refers to a protein determinant capable of binding to an antibody
of
the disclosure. Epitopes usually consist of chemically active surface
groupings of
molecules such as amino acids or sugar side chains and usually have specific
three dimensional structural characteristics, as well as specific charge
characteristics. Conformational and non-conformational epitopes are
distinguished in that the binding to the former but not the latter is lost in
the
presence of denaturing solvents. Such antibodies can be identified based on
their
ability to cross-compete (e.g., to competitively inhibit the binding of, in a
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
34
statistically significant manner) with antibodies such as 13H5, 13H7 or 7H9,
in
standard IFN-alpha binding assays. For example, as demonstrated in the
Examples of U.S. Patent No. 7,741,449, by Biacore analysis, 13H5 binds with
high affinity to IFN-alpha 2a and IFN-alpha 2b. Accordingly, in one
embodiment,
the disclosure provides antibodies, such as human antibodies, that compete for
binding to IFN-alpha 2a or IFN-alpha 2b with another antibody, such as13H5,
13H7 or 7H9. The ability of a test antibody to inhibit the binding of, e.g.,
13H5,
13H7 or 7H9 to IFN-alpha 2a or IFN-alpha 2b demonstrates that the test
antibody
can compete with that antibody for binding to IFN-alpha 2a or IFN-alpha 2b;
such
an antibody can, according to non-limiting theory, bind to the same or a
related
(e.g., a structurally similar or spatially proximal) epitope on IFN-alpha 2a
or IFN-
alpha 2b as the antibody with which it competes. In one embodiment, the
antibody that binds to the same epitope on IFN-alpha 2a or IFN-alpha 2b as,
e.g.,
13H5, 13H7, or 7H9, is a human monoclonal antibody. Such human monoclonal
antibodies can be used in the methods disclosed herein.
[0123] In one embodiment, the IFN-alpha antibody antagonist is sifalimumab.
Sifalimumab is a fully human, 147,000 Dalton IgGlk monoclonal antibody (Mab)
that selectively binds to multiple interferon-alpha subtypes. Sifalimumab is
made
from 100% human protein sequences, thereby making it a fully human
monoclonal antibody. Fully human monoclonal antibodies have advantages over
other forms of monoclonal antibodies, such as chimeric and humanized
antibodies, as they have a more favorable safety profile and can be eliminated
less rapidly from the human body, thereby reducing the frequency of dosing.
Sifalimumab was derived from the IgG4k antibody, 13H5 described above and in
U.S. Patent No. 7,741,449, which was selected based on functional assays as
having the most desirable properties for a potential therapeutic agent. 13H5
was
subsequently converted to an IgGI antibody isotype, produced in CHO cells. See
U.S. Patent No. 7,741,449, U.S. Patent Pub. Nos. 2010-0143372 and 2010-
0266610, and PCT Pub. Nos. W02008/070135, W02009/061818,
W02008/137838, W02008/070137, W02008/137835, W02009/155559,
W02008/121616, and W02008/121615, each of which is hereby incorporated
herein by reference.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
[0124] The nucleotide and amino acid sequences of the heavy chain variable
region of 13H5 are shown in SEQ ID NOs: 25 and 19, respectively.
[0125] The VH CDR1, CDR2, and CDR3 amino acid sequences of 13H5 are
shown in SEQ ID NOs: 1, 4, and 7, respectively.
[0126] The nucleotide and amino acid sequences of the light chain variable
region of 13H5 are shown in SEQ ID NOs: 28 and 22, respectively.
[0127] The VL CDR1, CDR2, and CDR3 amino acid sequences of 13H5 are
shown SEQ ID NOs: 10, 13, and 16, respectively.
Treatment and Dosing using Anti-IFN-alpha Antibodies
[0128] The disclosure includes methods of dosing to achieve a desired PK
characteristic. In a specific embodiment, the dose is chosen to achieve a
desired
characteristic chosen from Tmax (time of maximum observed concentration),
Tmax ss (time of maximum observed concentration at steady state), Cmax
(maximum observed concentration), Cmax ss (maximum observed concentration
at steady state), AUCT, (area under the curve over the dosing interval), AUCT
ss
(area under the curve over the dosing interval at steady state), Ctrough
(trough
observed concentration), Ctrough ss (trough observed concentration at steady
state), half-life (terminal elimination half-life, defined as In(2)/lambda z),
CLss
(serum steady state clearance, defined as Dose/AUCT ss), Vss (steady state
volume of distribution), AUClast (area under the curve from time 0 to last
observed concentration, i.e., Clast), AUCinf (area under the curve from 0 to
infinity, defined as (AUClast+Clast)/lambda z), AUCinf extrapolated (percent
of
the AUCinf curve extrapolated, defined as ((Clast/lambdaz)/AUCinf)*100), CL
(apparent total body clearance of the drug from plasma), CL/F (apparent serum
clearance), Vz/F (apparent terminal volume of distribution), CLss/F (apparent
serum steady state clearance), Vc (central volume), Vp (peripheral volume) or
Lambda z (slope of the terminal elimination phase).
[0129] In a specific embodiment, the methods of dosing produce a PK
parameter
of at least one of the values for the PK characteristics shown in Tables 2, 3,
6, 7,
and 8-11. The values of such desired PK characteristics are understood to
encompass the CV% as shown in the tables. In a specific embodiment, the
methods of the disclosure produce a value chosen from the PK characteristics
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
36
shown in Tables 2, 3, 6, 7, and 8-11 and neutralization of a the 21-gene or 4-
gene PD marker discussed herein. In a specific embodiment, the methods of the
disclosure produce a value chosen from the PK characteristics shown in Tables
2, 3, 6, 7, and 8-11, a neutralization of a the 21-gene or 4-gene PD marker
discussed herein, and a reduction in symptoms of SLE, such as a reduction in
SLEDAI score.
[0130] The term "dosage form" refers to a pharmaceutical composition
comprising
one or more active pharmaceutical ingredients (API), e.g., an anti-IFN-alpha
antibody or antigen-binding fragment thereof, such as sifalimumab, the
composition optionally containing pharmacologically inactive ingredients,
i.e.,
pharmaceutically acceptable carriers, fillers, excipients or combinations
thereof
such as polymers, suspending agents, surfactants, disintegrants, dissolution
modulating components, binders, fillers, lubricants, glidants, stabilizers,
antioxidants, osmotic agents, colorants, plasticizers, coatings and the like,
that
are used to manufacture and deliver active pharmaceutical agents. Examples of
pharmaceutical compositions comprising anti-interferon alpha antibodies can be
found in U.S. Patent Application Publication No. 2010-0209434 Al entitled
"Antibody Formulation", which is hereby incorporated by reference herein in
its
entirety.
[0131] An anti-IFN-alpha antibody or antigen-binding fragment thereof,
e.g.,
sifalimumab, can be administered to a human patient in accord with known
methods, such as intravenous administration, e.g., as a bolus or by continuous
infusion over a period of time, by intramuscular, intraperitoneal,
intracerobrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal,
oral,
topical, or inhalation routes, or a combination of two or more recited routes.
[0132] In one embodiment, a formulation comprising an anti-IFN-alpha
antibody
or antigen-binding fragment thereof, such as sifalimumab for use in the
methods
of the disclosure is for parenteral administration. In one embodiment, a
formulation of the disclosure comprising an anti-IFN-alpha antibody or antigen-
binding fragment thereof, such as sifalimumab is an injectable formulation. In
one
embodiment, a formulation of the disclosure comprising an anti-IFN-alpha
antibody or antigen-binding fragment thereof, such as sifalimumab is for
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
37
intravenous, subcutaneous, or intramuscular administration. In a specific
embodiment, a formulation of the disclosure comprises an anti-IFN-alpha
antibody or antigen-binding fragment thereof, such as sifalimumab wherein said
formulation is for subcutaneous injection.
[0133] As used herein the term "intravenous administration" refers to the
introduction of a composition to a patient into a vein. An anti-IFN-alpha
antibody
or antigen-binding fragment thereof can be administered intravenously (IV),
e.g.,
as an intravenous infusion or as an intravenous bolus. The term "intravenous
infusion" refers to introduction of a drug, e.g., an anti-IFN-alpha antibody
or
antigen-binding fragment thereof, such as sifalimumab, into the vein of an
animal
or human patient over a period of time greater than approximately 5 minutes,
for
example, between approximately 30 to 90 minutes, although, according to the
disclosure, intravenous infusion is alternatively administered for 10 hours or
less.
In one particular embodiment, the duration of the infusion is at least 60
minutes.
[0134] The term "intravenous bolus" or "intravenous push" refers to drug
administration, e.g., of an anti-IFN-alpha antibody or antigen-binding
fragment
thereof, such as sifalimumab, into a vein of an animal or human such that the
body receives the drug in approximately 15 minutes or less, for example, 5
minutes or less.
[0135] The term "subcutaneous administration" refers to introduction of a
drug,
e.g., an anti-IFN-alpha antibody or antigen-binding fragment thereof, such as
sifalimumab, under the skin of an animal or human patient, for example, within
a
pocket between the skin and underlying tissue, by relatively slow, sustained
delivery from a drug receptacle. The pocket can be created by pinching or
drawing the skin up and away from underlying tissue. In some embodiments, a
composition comprising an anti-IFN-alpha antibody or antigen-binding fragment
thereof, such as sifalimumab is introduced under the surface of the skin of
the
patient with a hypodermic needle.
[0136] The term "subcutaneous infusion" refers to introduction of a drug,
e.g., an
anti-IFN-alpha antibody or antigen-binding fragment thereof, such as
sifalimumab, under the skin of an animal or human patient, for example, within
a
pocket between the skin and underlying tissue, by relatively slow, sustained
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
38
delivery from a drug receptacle for a period of time including, but not
limited to,
30 minutes or less, or 90 minutes or less. Optionally, the infusion can be
made by
subcutaneous implantation of a drug delivery pump implanted under the skin of
the animal or human patient, wherein the pump delivers a predetermined amount
of drug for a predetermined period of time, such as 30 minutes, 90 minutes, or
a
time period spanning the length of the treatment regimen.
[0137] The term "subcutaneous bolus" refers to drug administration beneath
the
skin of an animal or human patient of an anti-IFN-alpha antibody or antigen-
binding fragment thereof, such as sifalimumab, where bolus drug delivery is
less
than about 15 minutes, less than about 5 minutes, or less than about 60
seconds.
Administration can be within a pocket between the skin and underlying tissue,
where the pocket is created, for example, by pinching or drawing the skin up
and
away from underlying tissue.
[0138] In one embodiment, a formulation for use in the methods of the
disclosure
is for aerosol administration.
Weight Based Dosage Administration
[0139] In some embodiments, the dosage of anti-IFN-alpha antibody or
antigen-
binding fragment thereof administered to a patient is calculated, e.g., as a
function of the patient body mass (weight), height, or body surface. In
certain
embodiments, the dosage of anti-IFN-alpha antibody or antigen-binding fragment
thereof administered to a patient depends on the patient's body weight. The
weight-based dose is generally provided in mg/kg.
[0140] In some embodiments, an anti-IFN-alpha antibody or antigen-binding
fragment thereof is administered at a weight-based dosage of about 0.1 mg/kg,
or
about 0.2 mg/kg, or about 0.3 mg/kg, or about 0.4 mg/kg, or about 0.5 mg/kg,
or
about 0.6 mg/kg, or about 0.7 mg/kg, or about 0.8 mg/kg, or about 0.9 mg/kg.
In
other embodiments, an anti-IFN-alpha antibody or antigen-binding fragment
hereof is administered at a weight-based dosage of about 1 mg/kg, or about 2
mg/kg, or about 3 mg/kg, or about 4 mg/kg, or about 5 mg/kg, or about 6 mg/kg,
or about 7 mg/kg, or about 8 mg/kg, or about 9 mg/kg. In certain embodiments,
an anti-IFN-alpha antibody or antigen-binding fragment thereof is administered
at
a weight-based dosage of about 10 mg/kg, or 15 mg/kg, or 20 mg/kg, or about 25
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
39
mg/kg, or about 30 mg/kg, or about 35 mg/kg, or about 40 mg/kg, or about 45
mg/kg, or about 50 mg/kg, or about 55 mg/kg, or about 60 mg/kg, or about 65
mg/kg, or about 70 mg/kg, or about 75 mg/kg, or about 80 mg/kg, or about 85
mg/kg, or about 90 mg/kg, or about 95 mg/kg, or about 100 mg/kg. In specific
embodiments, the anti-IFN-alpha antibody or antigen-binding fragment thereof
is
administered at a weight-base dosage about 0.3 mg/kg, or about 1.0 mg/kg, or
about 3.0 mg/kg, or about 10 mg/kg. In certain embodiments, the weight-based
dosage is administered intravenously. In other embodiments, the weight-based
dosage is administered subcutaneously. In some specific embodiments, the an
anti-IFN-alpha antibody is sifalimumab.
[0141] When a series of weight-based doses of an anti-IFN-alpha antibody or
antigen-binding fragment thereof, such as sifalimumab, are administered, these
doses can, for example, be administered approximately every week,
approximately every 2 weeks, approximately every 3, or about every 4 weeks. In
some embodiments, weight-based doses of an anti-IFN-alpha antibody or
antigen-binding fragment thereof are administered approximately every day,
approximately every two days, approximately every three days, approximately
every 4 days, approximately every 5 days, approximately every 6 days, or
approximately every seven days.
[0142] In a specific embodiment, weight-based doses of an anti-IFN-alpha
antibody or antigen-binding fragment are administered every 2 weeks. The
weight-based doses of anti-IFN-alpha antibody or an antigen-binding fragment
thereof can be administered, for example, for about 1 month, or about 2
months,
or about 3 months, or about 4 months, or about 5 months, or about 6 months.
The weight-based doses of anti-IFN-alpha antibody or an antigen-binding
fragment thereof, can, for example, continue to be administered until disease
progression, adverse event, or other parameter occurs as determined by the
physician. In a specific embodiment, weight-based doses of anti-IFN-alpha
antibody or an antigen-binding fragment thereof, are administered for about 6
months. In a more specific embodiment, weight-based doses anti-IFN-alpha
antibody or an antigen-binding fragment thereof, are administered for about 26
weeks.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
[0143] In
some embodiments, patients can be administered at least one, at least
2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at
least 9, at
least 10, at least 11, at least 12, at least 13, at least 14 or at least 15
weight-
based doses of an anti-IFN-alpha antibody or antigen-binding fragment thereof,
such as sifalimumab. In a specific embodiment, patients are administered at
least
14 doses of an anti-IFN-alpha antibody or antigen-binding fragment thereof,
such
as sifalimumab. In some specific embodiments, patients can be administered at
least 14 IV weight-based doses of an anti-IFN-alpha antibody or antigen-
binding
fragment thereof, such as sifalimumab.
[0144] In some embodiments, weight-based doses of anti-IFN-alpha
antibody or
an antigen-binding fragment thereof, are administered at equal time intervals.
In
other embodiment, such weight-based doses are administered at varying
intervals. In
some embodiments, all administered weight-based doses as
essentially identical. In other embodiments, at least one weight-based dose is
different with respect to the other doses, e.g., in volume, concentration,
route of
administration, formulation, etc.
Fixed Dosage Administration
[0145] A
"fixed dose" or "fixed dosage" of a therapeutic agent herein refers to a
dose that is administered to a human patient without regard for the weight
(WT)
or body surface area (BSA) of the patient. The fixed dose of anti-IFN-alpha
antibody or an antigen-binding fragment thereof, e.g., sifalimumab, is
therefore
not provided as a mg/kg dose or mg/m2 dose, but rather as an absolute amount
of the therapeutic agent.
[0146] In some embodiments, an anti-IFN-alpha antibody or antigen-
binding
fragment thereof is administered at a fixed dosage of about 10 mg, or about 20
mg, or about 30 mg, or about 40 mg, or about 50 mg, or about 60 mg, or about
70 mg, or about 80 mg, or about 90 mg, or about 100 mg. In other embodiments,
an anti-IFN-alpha antibody or antigen-binding fragment thereof is administered
at
a fixed dosage of about 100 mg, or about 150 mg, about 200 mg, or about 300
mg, or about 400 mg, or about 500 mg, or about 600 mg, or about 700 mg, or
about 800 mg, or about 900 mg, or about 100 mg, or about 1100 mg, or about
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
41
1200 mg, or about 1300 mg, or about 1400 mg, or about 1500 mg, or about 1600
mg, or about 1700 mg, or about 1800 mg, or about 1900 mg, or about 2000 mg.
[0147] In specific embodiments, the anti-IFN-alpha antibody or antigen-
binding
fragment thereof is administered intravenously at a fixed dosage about 100 mg,
or about 150 mg, or about 200 mg, or about 600 mg, or about 1200 mg. In a
specific embodiment, the anti-IFN antibody or antigen-binding fragment thereof
is
administered subcutaneously at a fixed dosage of about 100 mg, or about 200
mg, or about 600 mg, or about 1200 mg. In some embodiments, a loading dose is
administered. In some specific embodiments, the anti-IFN-alpha antibody is
sifalimumab, or an antigen-binding fragment thereof. In a specific embodiment,
the anti-IFN-alpha antibody or antigen-binding fragment thereof is
administered
intravenously at a fixed dosage about 100 mg, or about 150 mg, or about 200
mg, or about 600 mg, or about 1200 mg once per month, with loading dose at
Day 14.
[0148] When a series of fixed doses of an anti-IFN-alpha antibody or
antigen-
binding fragment thereof are administered, these doses can, for example, be
administered approximately every week, approximately every 2 weeks,
approximately every 3, or about every 4 weeks. In some embodiments, fixed
doses of an anti-IFN-alpha antibody or antigen-binding fragment thereof are
administered approximately every day, approximately every two days,
approximately every three days, approximately every 4 days, approximately
every 5 days, approximately every 6 days, or approximately every seven days.
In
a specific embodiment, the fixed dose of anti-IFN-alpha antibody or antigen-
binding fragment thereof is a 100 mg dose administered daily. In a specific
embodiment, the fixed dose of anti-IFN-alpha antibody or antigen-binding
fragment thereof is a 150 mg dose administered daily. In specific embodiments,
the fixed dose of anti-IFN-alpha antibody is a 100 mg daily dose or a 150 mg
daily dose of sifalimumab.
[0149] In a specific embodiment, fixed doses of anti-IFN-alpha antibody or
antigen-binding fragment thereof are administered every 2 weeks. These fixed
doses can be administered, for example, for about 1 month, or about 2 months,
or about 3 months, or about 4 months, or about 5 months, or about 6 months.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
42
Such fixed doses can, for example, continue to be administered until disease
progression, adverse event, or other parameter occurs as determined by the
physician.. In some embodiments, patients can be administered at least one, at
least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least
8, at least 9,
at least 10, at least 11, at least 12, at least 13, at least 14 or at least 15
fixed
doses of anti-IFN-alpha antibody or antigen-binding fragment thereof. In a
specific embodiment, patients are administered at least 13 doses of anti-IFN-
alpha antibody or antigen-binding fragment thereof. In some specific
embodiments, patients can be administered at least 13 subcutaneous fixed doses
of sifalimumab.
[0150] In some embodiments, fixed doses of anti-IFN-alpha antibody or
antigen-
binding fragment thereof are administered at equal time intervals. In other
embodiment, fixed doses of anti-IFN-alpha antibody or antigen-binding fragment
thereof are administered at varying intervals. In some embodiments, all
administered fixed doses are essentially identical. In other embodiments, at
least
one fixed dose is different with respect to the other doses, e.g., in volume,
concentration, route of administration, formulation, etc.
[0151] For the prevention or treatment of autoimmune diseases, e.g., SLE,
scleroderma, or myositis, the fixed dose or weight-based of anti-IFN-alpha
antibody, e.g., sifalimumab, or antigen-binding fragment thereof will depend
on
the type of disease to be treated, as defined above, the severity and course
of
the disease, whether the antibody is administered for preventive or
therapeutic
purposes, previous therapy, the patient's clinical history and response to the
antibody, and the discretion of the attending physician.
[0152] In some embodiments, one or more loading dose(s) of the anti-IFN-
alpha
antibody or antigen-binding fragment thereof, either weigh-based or fixed
doses,
can be administered, followed by one or more maintenance dose(s), either
weigh-based or fixed doses, of the antibody. In another embodiment, a
plurality of
the same fixed doses of anti-IFN-alpha antibody or antigen-binding fragment
thereof are administered to the patient. According to one embodiment of the
disclosure, one or more weigh-based or fixed loading doses of anti-IFN-alpha
antibody (e.g. sifalimumab) or an antigen-binding fragment can be
administered,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
43
followed by one or more weigh-based or fixed maintenance doses of the
antibody. According to another embodiment of the disclosure, one or more weigh-
based or fixed dose(s) of anti-IFN-alpha antibody (e.g. sifalimumab) or an
antigen-binding fragment can be administered for up a number of cycles. In
another embodiment, a weigh-based or fixed dose of anti-IFN-alpha antibody
(e.g., sifalimumab) or an antigen-binding fragment can be administered as a
loading dose, followed by one or more maintenance dose(s). About one, two or
more maintenance doses of anti-IFN-alpha antibody or antigen-binding fragment
thereof can be administered to the patient according to this embodiment.
[0153] Thus, the disclosure provides a method of treating an autoimmune
disorder, e.g., SLE, scleroderma, or myositis in a human patient comprising
administering at least one fixed dose of sifalimumab to the patient, wherein
the
fixed dose is about 100 mg, about 200 mg, about 600 mg, or about 1200 mg of
sifalimumab.
Dosing to Achieve Desired Pharmacokinetic Characteristics
[0154] The term "pharmacokinetic characteristics" or PK characteristics
refers to
parameters describing the mechanisms of absorption and distribution of an
administered drug, e.g., an anti-IFN-alpha antibody such as sifalimumab or an
antigen-binding fragment thereof, the rate at which a drug action begins and
the
duration of the effect, the physicochemical chemical changes of the substance
in
the body and the effects and routes of excretion of the metabolites of the
drug.
The methods disclosed herein, permit dose selection to achieve desired PK
characteristics, whether the dosing be based on weight, fixed dosing, and
whether the route of administration is, for example, intravenous or
subcutaneous.
[0155] Such pharmacokinetic characteristics comprise, e.g., T. (time of
maximum observed concentration), Tmax ss (time of maximum observed
concentration at steady state), C. (maximum observed concentration), Cmax ss
(maximum observed concentration at steady state), AUCT, (area under the curve
over the dosing interval), AUCTss (area under the curve over the dosing
interval at
steady state), Ctrough (trough observed concentration), Ctrough ss (trough
observed
concentration at steady state), half-life (terminal elimination half-life,
defined as
In(2)6,,), CLss (serum steady state clearance, defined as Dose/AUCT ss), Vss
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
44
(steady state volume of distribution), AUCiast (area under the curve from time
0 to
last observed concentration, i.e., Clast), AUC,pf (area under the curve from 0
to
infinity, defined as (AUCIast+Clast)6,,), AUC,pf extrapolated (percent of the
AUC,pf
curve extrapolated, defined as ((Ciastaz)/AUC,pf)*100), CL (apparent total
body
clearance of the drug from plasma), CL/F (apparent serum clearance), Vz/F
(apparent terminal volume of distribution), CLss/F (apparent serum steady
state
clearance), Vc (central volume), Vp (peripheral volume) or 2 (slope of the
terminal
elimination phase).
[0156] Pharmacokinetic characteristics can be used to determine the
appropriate
dosage of a drug of interest, e.g., an anti-IFN-alpha antibody or antigen-
binding
fragment thereof. Characteristics describing the blood plasma curve can be
obtained in clinical trials by administration of the active agent to a number
of test
subjects. The blood plasma values of the individual test persons are then
averaged.
[0157] In the context of the present disclosure, pharmacokinetic
characteristics,
e.g., AUC, C. and Tmax refer to mean values corresponding to a population of
subjects. Further, in the context of the present disclosure, in vivo
parameters
such as values for AUC, Cmax, Tmax refer to parameters or values obtained
after
administration at steady state to human patients.
[0158] To quantify pharmacokinetic characteristics in patients, the patient
group
comprises between 10 to 200 patients. A reasonable number of patients is,
e.g.,
10, 20, 30, 40, 50, 75, 100, 125, 150, 175 or 200 patients. Patients are be
selected according to inclusion and exclusion criteria related to the ability
of a
physician to adequately discern the symptoms of the condition to be treated
and
the effect of the tested drug over those symptoms.
[0159] The calculation of pharmacokinetic characteristics can be performed
with
the WinNonlin program in any of its versions and/or platform implementations.
The skilled artisan will understand that such calculations can also be
performed
using other available software packages or by manual calculation according to
formulas and methods known in the art.
[0160] For clarity and convenience herein, the convention is utilized of
designating the time of drug administration or initiation of testing as zero
hours
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
(t=0 hours) or zero days (Day 0) and times following administration in
appropriate
time units, e.g., t=30 minutes or Day 68.
[0161] The term "bioavailability" is defined for purposes of the present
disclosure
as the extent to which an active agent such as an anti-IFN-alpha antibody,
e.g.,
sifalimumab, or an antigen-binding fragment thereof is absorbed from the unit
dosage forms. AUC provides a measure of bioavailability.
[0162] The term "steady state" means that a plasma level for a given drug,
e.g.,
an anti-INF-alpha antibody, e.g., sifalimumab, or an antigen binding fragment
thereof, has been achieved and which is maintained with subsequent doses of
the drug at a level which is at or above the minimum effective therapeutic
level
and is below the minimum toxic plasma level. It will be well understood by
those
skilled in the medical art that after the administration of each dose the
concentration passes through a maximum and then again drops to a minimum.
Accordingly, the steady state can be described as follows: At the time t=0,
the
time the first dose is administered, the concentration C is also O. The
concentration then passes through a first maximum and then drops to a first
minimum. Before the concentration drops to 0, another dose is administered, so
that the second increase in concentration doesn't start at O. Building on this
first
concentration minimum, the curve passes through a second maximum after the
second dose has been administered, which is above the first maximum, and
drops to a second minimum, which is above the first minimum. Thus, the blood
plasma curve escalates due to the repeated doses and the associated step-by-
step accumulation of active agent, until it levels off to a point where dose
administered and elimination are in balance. This state, at which dose
administered and elimination are in equilibrium and the concentration
oscillates
constantly between a defined minimum and a defined maximum, is called steady
state.
[0163] As used herein, the term "clearance rate" refers to CL (apparent
total body
clearance of the drug from plasma),CLõ ( serum steady state clearance), CL/F
(apparent serum clearance) and CLõ/F (apparent serum steady state clearance).
As used herein, term "apparent volume of distribution" refers to Võ (steady
state
volume of distribution) and Vz/F (apparent terminal volume of distribution).
As
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
46
used herein, the term "half-life" refers to a biological half-life of a
particular
binding agent (e.g., an anti-INF-alpha antibody, such as sifalimumab, or an
antigen binding fragment thereof) in vivo. Half-life can be represented by the
time required for half the quantity administered to a subject to be cleared
from the
circulation and/or other tissues in the subject.
[0164] In some embodiments of the present disclosure, an anti-IFN-alpha or
antigen-binding fragment thereof is administered in a dosage, such that an
effective exposure is provided in a patient, for example as measured by, e.g.,
clearance rate, apparent volume of distribution, or half-life. The disclosure
also
includes methods which combine achieving two or more favorable
pharmacokinetic characteristics or combinations thereof. Examples of those
pharmacokinetic parameters include clearance rate clearance rate (CL, CLõ,
CL/F, or CLss/F), an apparent volume of distribution (Võ or Vz/F), and a serum
half-life. For instance, the formulation administered through the methods of
the
disclosure can be selected such that when administered to a patient in need
thereof, the selected formulation provides the patient with one or more of the
desired pharmacokinetic characteristics.
[0165] In some embodiments, one or more desired pharmacokinetic
characteristics is achieved after the administration of one or more doses of
an
anti-INF antibody or an antigen binding fragment thereof to a subject
suffering
from an autoimmune disorder. In some embodiments, such interferon alpha is
human interferon alpha. In other embodiments, the one or more desired
pharmacokinetic characteristics are selected, e.g., from the group consisting
of
from the group consisting of a clearance rate (CL, CLõ, CL/F, or CLõ/F),
apparent volume of distribution (Vss or Vz/F), and a serum half-life. In some
embodiments, the immune disorder is systemic lupus erythematosus,
scleroderma, or myositis.
[0166] In some embodiments, the desired pharmacokinetic characteristics are
selected from the group consisting for a clearance rate (CL, CLss, CL/F, or
CLss/F) of between about 99 and about 432 mL/day, an apparent volume of
distribution (Vss or Vz/F) of between about 3 and about 17 L, and a serum half-
life
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
47
of about 14 days to about 47 days is achieved following the administration of
the
anti-IFN antibody or antigen-binding fragment thereof.
[0167] In some embodiments, the desired clearance rate (CL, CLõ, CL/F, or
CL/F) is about 90, about 100, about 110, about 120, about 130, about 140,
about 150, about 160, about 170, about 180, about 190, about 200, about 210,
about 220, about 230, about 240, about 250, about 260, about 270, about 280,
about 290, about 300, about 310, about 320, about 330, about 340, about 350,
about 360, about 370, about 380, about 390, about 400, about 410, about 420,
about 430, or about 440 mL/day. .
[0168] In some embodiments the desired apparent volume of distribution (Võ
or
Vz/F) of about 3, about 4, about 5, about 6, about 7, about 8, about 9, about
10,
about 11, about 12, about 13, about 14, about 15, about 16, or about 17 L. In
some embodiments, the desired serum half-life is about 10, about 11, about 12,
about 13, about 14, about 15, about 16, about 17, about 18, about 19, about
20,
about 21, about 22, about 23, about 24, about 25, about 26, about 27, about
28,
about 29, about 30, about 31, about 32, about 33, about 34, about 35, about
36,
about 37, about 38, about 39, about 40, about 41, about 42, about 43, about
44,
about 45, about 46, or about 47 days.
[0169] In some embodiments, such desired pharmacokinetic characteristics
are
achieved after the administration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, or
15 doses. In some embodiments, such desired pharmacokinetic characteristics
are achieved after the administration of at least 15 doses of an anti-IFN-
alpha
antibody, e.g., sifalimumab, or an antigen-binding fragment thereof. In some
embodiments, such doses are intravenous. In other embodiments, such doses
are subcutaneous. In other embodiments, such doses are weight-based, whereas
in other cases such doses are fixed doses.
[0170] When fixed doses are administered to achieve the desired
pharmacokinetic characteristics, such doses an anti-IFN-alpha antibody or
antigen-binding fragment thereof can be of about 10 mg, about 20 mg, about 30
mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about
90 mg, or about 100 mg. In other embodiments, an anti-IFN-alpha antibody or
antigen-binding fragment thereof is administered at a fixed dosage of about
100
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
48
mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg,
about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about
100 mg, about 1100 mg, about 1200 mg, about 1300 mg, about 1400 mg, about
1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900 mg, about
2000 mg. In some embodiments, a loading dose is administered.
[0171] In a specific embodiment, the anti-IFN-alpha antibody or antigen-
binding
fragment thereof is administered intravenously at a fixed dosage about 100 mg,
or about 150 mg, or about 200 mg, or about 250 mg, or about 300 mg, or about
400 mg, or about 500 mg, or about 600 mg, or about 700 mg, or about 800 mg,
or about 900 mg, or about 1000 mg, or about 1100 mg, or about 1200 mg, or
about 1300 mg, or about 1400 mg, or about 1500 mg, or about 1600 mg, or about
1700 mg, or about 1800 mg, or about 1900 mg, or about 2000 mg once per
month, with a loading dose at Day 14.
[0172] In some embodiments, the desired pharmacokinetic characteristics can
be
achieved by administering doses of an anti-IFN-alpha antibody or antigen-
binding
fragment thereof approximately every day, approximately every two days,
approximately every three days, approximately every 4 days, approximately
every 5 days or approximately every 6 days. In some embodiments, the desired
pharmacokinetic characteristics can be achieved by administering doses of an
anti-IFN-alpha antibody or antigen-binding fragment thereof approximately
every
week, approximately every 2 weeks, approximately every 3 weeks, or about
every 4 weeks. In a specific embodiment, weight-based doses of anti-IFN-alpha
antibody or antigen-binding fragment thereof are administered every 2 weeks.
In
other embodiments, the desired pharmacokinetic characteristics can be achieved
by administering doses of an anti-IFN-alpha antibody or antigen binding
fragment
thereof, for example, for about 1 month, or about 2 months, or about 3 months,
or
about 4 months, or about 5 months, or about 6 months.
[0173] In some embodiments, the time to reach maximum plasma concentration
(Tmax or Tmax ss) following IV administration of an anti-IFN-alpha, such as
sifalimumab, or an antigen-binding fragment thereof is about 0.13 days or
less.
[0174] In some embodiments, the administration of an anti-IFN-alpha, such
as
sifalimumab, or an antigen-binding fragment thereof achieves a desired
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
49
pharmacokinetic characteristics chosen from clearance rate (CL, CLss, CL/F, or
CLõ/F), apparent volume of distribution (Võ or Vz/F), a serum half-life, T.
(time
of maximum observed concentration), Tmax ss (time of maximum observed
concentration at steady state), C. (maximum observed concentration), Cmax ss
(maximum observed concentration at steady state), AUCT, (area under the curve
over the dosing interval), AUCTss (area under the curve over the dosing
interval at
steady state), Ctrough (trough observed concentration), Ctrough ss (trough
observed
concentration at steady state), half-life (terminal elimination half-life,
defined as
In(2)6), CL ss (serum steady state clearance, defined as Dose/AUCT ss), Vss
(steady state volume of distribution), AUClast (area under the curve from time
0 to
last observed concentration, i.e., Clast), AUC,nf (area under the curve from 0
to
infinity, defined as (AUClast+Clast)6,,), AUC,nf extrapolated (percent of the
AUC,nf
curve extrapolated, defined as ((Ciastaz)/AUC,nf)*100), CL/F (apparent serum
clearance), Vz/F (apparent terminal volume of distribution), CLss/F (apparent
serum steady state clearance), Vc (central volume), Vp (peripheral volume),
(slope of the terminal elimination phase), or combinations thereof.
[0175] In some embodiments a single IV administration of about 0.3 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tmax of
about 0.12 days or less, a maximum plasma concentration (C.) of about 7,
about 8, about 9, about 10, about 11, about 12, about 13, about 14 or about 15
pg/mL, an area under the plasma concentration-time curve during a dosage
interval (T) (AUCT) of about 50, about 55, about 60, about 65, about 70, about
75,
about 80, about 85, about 90, about 95, about 100, about 105, or about 110 pg
day/mL, and a trough plasma concentration (Ctrough) of about 2.0, about 2.2,
about 2.4, about 2.6, about 2.8, about 3.0, about 3.2, about 3.4, about 3.6,
about
3.8, or about 4.0 pg/mL.
[0176] In some embodiments, a single IV administration of about 0.3 mg/kg
to a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from: an average T. of about 0.07 days, an average C. of about 11
pg/mL, an average AUCT of about 79 pg day/mL, and an average Ctrough of about
3 pg/mL.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
[0177] In some embodiments, a single IV administration of about 1 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tri,õ of
about 0.12 days or less, a Crnax of about 20, about 25, about 30, about 35,
about
40, or about 45 pg/mL, an AUCT of about 150, about 175, about 200, about 225,
about 250, about 275, or about 300 pg day/mL, and a Ctrough of about 4, about
5,
about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13,
about, or about 15 pg/mL .
[0178] In some embodiments, a single IV administration of about 1 mg/kg to
a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from: an average Trnax of about 0.08 days, an average Crnax of about 32
pg/mL, an average AUCT of about 221 pg day/mL, and an average Ctrough of about
8 pg/mL.
[0179] In some embodiments, a single IV administration of about 3 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tri,õ of
about 0.13 days or less, a Crnax of about 60, about 70, about 80, about 90,
about
100, about 110, about 120, about 130, about 140, or about 150 pg/mL, an AUCT
of about 450, about 500, about 550, about 600, about 650, about 700, about
750,
about 800, about 850, about 900, about 950, about 1000, or about 1050 pg
day/mL, and a Ctrough of about 12, about 14, about 16, about 18, about 20,
about
22, about 24, about 26, about 28, about 30, about 32, about 34, or about 36
pg/mL.
[0180] In some embodiments, a single IV administration of about 3 mg/kg to
a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from: an average Trnax of about 0.09 days, an average Crnax of about
103
pg/mL, an average AUCT of about 739 pg day/mL, and an average Ctrough of about
23 pg/mL.
[0181] In some embodiments, a single IV administration of about 10 mg/kg
achieves one or more pharmacokinetic characteristics chosen from: a Tri,õ of
about 0.13 days or less, a Crnax of about 140, about 150, about 160, about
170,
about 180, about 190, about 200, about 210, about 220, about 230, about 240,
about 250, about 260, about 270, about 280, about 290, about 300, about 310,
or
about 320 pg/mL, an AUCT of about 900, about 1000, about 1100, about 1200,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
51
about 1300, about 1400, about 1500, about 1600, about 1700, about 1800, about
1900, about 2000, about 2100, about 2200, or about 2300 pg day/mL, and a
Ctrough Of about 25, about 30, about 35, about 40, about 45, about 50, about
55,
about 60, about 65, about 70, about 75, or about 80 pg/mL.
[0182] In some embodiments, a single IV administration of about 10 mg/kg to
a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from: an average T. of about 0.09 days, an average C. of about 230
pg/mL, an average AUCT of about 1610 pg day/mL, and an average Ctrough Of
about 52 pg/mL.
[0183] In some embodiments, a sufficient number of IV doses of about 0.3
mg/kg
are administered at about 14-day intervals to achieve a steady state, and
wherein
one or more steady state pharmacokinetic characteristics are chosen from: a T.
ss of about 0.60 days or less, a Cmax ss of about 11, about 12, about 13,
about 14,
about 15, about 16, about 17, about 18, about 19, about 20, about 21, about
22,
about 23, about 24, or about 25 pg/mL, an AUCT ss of about 80, about 90 ,
about
100, about 110, about 120, about 130, about 140, about 150, about 160, about
170, about 180, about 190, or about 200 pg day/mL, and a Ctrough ss of about 5
of
less, about 6, about 7, about 8, about 9, about 10, or about 11 pg/mL is
achieved.
[0184] In some embodiments, a sufficient number of IV doses of about 0.3
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics chosen
from: an average T. of about 0.17 days, an average C. of about 18 pg/mL, an
average AUCT of about 143 pg day/mL, and an average Ctrough Of about 8 pg/mL
is achieved.
[0185] In some embodiments, a sufficient number of IV doses of about 0.3
mg/kg
are administered at about 14-day intervals to a achieve a steady state, and
wherein one or more pharmacokinetic characteristics selected from the group
consisting of a clearance rate (CLss) of about 90, about 100, about 110, about
120, about 130, about 140, about 150, about 160, about 170, about 180, about
190, about 200, about 210, about 220, about 230, about 240, about 240, about
250, about 260, or about 270 mL/day, an apparent volume of distribution (Vss)
of
about 4, about 5, about 6, about 7, about 8, or about 9 L, and a serum half-
life of
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
52
about 15 days, about 20 days, about 25 days, about 30 days, about 35 days,
about 40 days, or about to about 45 days is achieved.
[0186] In some embodiments, a sufficient number of IV doses of about 0.3
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics
selected from the group consisting of an average clearance rate (CLõ) of about
185 mL/day, an average apparent volume of distribution (Vss) of about 6 L, and
an average serum half-life of about 29 days is achieved.
[0187] In some embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered at about 14-day intervals to achieve a steady state, and
wherein
one or more steady state pharmacokinetic characteristics chosen from: a Tmax
ss
of about 0.11 days or less, a Cmax ss of about 25, about 30, about 35, about
40,
about 45, about 50, about 55, about 60, about 65, or about 70 pg/mL, an AUCT
ss
of about 200, about 250, about 300, about 350, about 400, about 450, about
500,
about 550, or about 600 pg day/mL, and a Ctrough ss of about 9, about 11,
about
13, about 15, about 17, about 19, about 21, about 23, about 25, about 27,
about
29, or about 31 pg/mL is achieved.
[0188] In some embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics chosen
from: an average Tmax ss of about 0.07 days, an average Cmax ss of about 48
pg/mL, an average AUCT ss of about 197 pg day/mL, and an average Ctrough ss of
about 11 pg/mL is achieved.
[0189] In some embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered at about 14-day intervals to a achieve a steady state, and
wherein one or more pharmacokinetic characteristics selected from the group
consisting of a clearance rate (CLss) of about 120, about 140, about 160,
about
180, about 200, about 220, about 240, about 260, about 280, about 300, about
320, about 340, or about 360 mL/day, an apparent volume of distribution (Vss)
of
about 4, about 5, about 6, about 7, about 8, or about 9 L, and a serum half-
life of
about 15, about 16, about 17, about 18, about 19, about 20, about 21, about
22,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
53
about 23, about 24, about 25, about 26, about 27, about 28, about 29, about
30,
about 31, or about 32 days is achieved.
[0190] In some embodiments, a sufficient number of IV doses of about 1
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics
selected from the group consisting of an average clearance rate (CLõ) of about
223 mL/day, an average apparent volume of distribution (Vss) of about 6 L, and
an average serum half-life of about 23 days is achieved.
[0191] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered at about 14-day intervals to achieve a steady state, and
wherein
one or more steady state pharmacokinetic characteristics chosen from: a Tmax
ss
of about 0.35 days or less, a C max ss of about 75, about 100, about 125,
about
150, about 175, about 200, about 225, or about 250 pg/mL, an AUCT ss of about
500, about 600, about 700, about 800, about 900, about 1000, about 1100 ,
about 1200, about 1300, about 1400, about 1500, about 1600, about 1700, about
1800, or about 1900 pg day/mL, and a Ctrough ss of about 25, about 30, about
35,
about 40, about 45, about 50, about 55, about 60, about 65, about 70, or about
75 pg/mL is achieved.
[0192] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics chosen
from: an average Tmax ss of about 0.13 days, an average Cmax ss of about 153
pg/mL, an average AUCT ss of about 1188 pg day/mL, and an average Ctrough ss
of
about 50 pg/mL is achieved.
[0193] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered at about 14-day intervals to a achieve a steady state, and
wherein one or more pharmacokinetic characteristics selected from the group
consisting of a clearance rate (CLss) of about 130, about 140, about 150,
about
160, about 170, about 180, about 190, about 200, about 210, about 220, about
230, about 240, about 250, about 260, about 270, about 280, about 290, about
300, or about 310 mL/day, an apparent volume of distribution (Vss) of about
3.0,
about 4.0, about 4.5, about 5.0, about 5.5, about 6.0, about 6.5, or about 7.0
L,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
54
and a serum half-life of about 14 days, about 15, about 16 days, about 17
days,
about 18 days, about 19 days, about 20 days, about 21 days, about 22 days,
about 23 days. about 24 days, about 25 days, or about 26 days is achieved.
[0194] In some embodiments, a sufficient number of IV doses of about 3
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics
selected from the group consisting of an average clearance rate (CLõ) of about
220 mL/day, an average apparent volume of distribution (Vss) of about 5 L, and
an average serum half-life of about 20 days is achieved.
[0195] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered at about 14-day intervals to achieve a steady state, and
wherein
one or more steady state pharmacokinetic characteristics chosen from: a Tmax
ss
of about 0.85 days or less, a Cmax ss of about 275, about 300, about 325,
about
350, about 375, about 400, about 425, about 450, about 475, about 500, about
525, about 550, about 575, or about 600 pg/mL, an AUCT ss of about 2500, about
2600, about 2700, about 2800, about 2900, about 3000, about 3100, about 3200,
about 3300, about 3400, about 3500, about 3600, about 3700, about 3800, about
3900, about 4000, about 4100, about 4200, or about 4300 pg day/mL, and a
Ctrough ss of about 90, about 100, about 110, about 120, about 130, about 140,
about 150, about 160, about 170, about 180, about 190, about 200, about 210,
about 220, about 230, about 240, about 250, about 260, about 270, about 280,
or
about 290 pg/mL is achieved.
[0196] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics chosen
from: an average Tmax ss of about 0.23 days, an average Cmax ss of about 232
pg/mL, an average AUCT ss of about 3403 pg day/mL, and an average Ctrough ss
of
about 184 pg/mL is achieved.
[0197] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered at about 14-day intervals to a achieve a steady state, and
wherein one or more pharmacokinetic characteristics selected from the group
consisting of a clearance rate (CLss) of about 150, about 160, about 170,
about
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
180, about 190, about 200, about 210, about 220, about 230, about 240, about
250, about 260, about 270, about 280, about 290, about 300, about 310, or
about
320 mL/day, an apparent volume of distribution (Võ) of about 4.0, about 4.5,
about 5.0, about 5.5, about 6.0, about 6.5, or about 7 L, and a serum half-
life of
about 15 days, about 16 days, about 17 days, about 18 days, about 19 days,
about 20 days, about 21 days, about 22 days, about 23 days, about 24 days,
about 25 days, about 26 days, about 27 days, about 28 days, or about 29 days
is
achieved.
[0198] In some embodiments, a sufficient number of IV doses of about 10
mg/kg
are administered to a population of subjects at about 14-day intervals to
achieve
a steady state, and wherein one or more pharmacokinetic characteristics chosen
from an average clearance rate (CLõ) of about 238 mL/day, an average apparent
volume of distribution (Vss) of about 6 L, and an average serum half-life of
about
22 days is achieved.
[0199] In some embodiments, the number of IV doses at about 14-day
intervals
required to achieve steady state is about 5 to about 8 doses. In some
embodiments, the desired pharmacokinetic characteristics are achieved after
the
antibody or antigen-binding fragment thereof is administered as a single dose
or
is administered in two or more doses once per week, once every two weeks,
once every three weeks, once every four weeks, once a month, once every 3
months, once every six months, or at varying intervals. In some embodiments,
the desired pharmacokinetic parameters are achieved after subcutaneous (SC)
administration.
[0200] In some embodiments, the desired pharmacokinetic characteristics are
achieved after the administration of 100 mg of anti-IFN-alpha administered as
a
single dose, or administered weekly, bi-weekly, or monthly. In some
embodiments, the desired pharmacokinetic characteristics are achieved after
the
administration of 150 mg of anti-IFN-alpha administered as a single dose, or
administered weekly, bi-weekly, or monthly. In some embodiments, the dose is
administered intravenously. In other embodiments the dose is administered
subcutaneously.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
56
[0201] In some embodiments, at T. or Tmax ss of between about 1.8 and about
9.4 days is achieved. In some embodiments the T. or Tmax ss is about 2 days or
lower, about 3 days or lower, about 4 days or lower, about 5 days or lower,
about
6 days or lower, about 7 days or lower, about 8 days or lower, about 9 days or
lower, or about 10 days or lower.
[0202] In some embodiments, a single SC administration of about 100 mg
achieves one or more pharmacokinetic characteristics chosen from: a Tmax of
about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or
about
days, a Cmax of about 4, about 5, about 6, about 7, about 8, about 9, about
10,
about 11, about 12, about 13, about 14, about 15, about 16, about 17, about
18,
about 19, about 20, or about 21 pg/mL, an area under the plasma concentration-
time curve from time zero to time of last measurable concentration (AUCiast)
of
about 175, about 200, about 225, about 250, about 275, about 300, about 325,
about 350, about 375, about 400, about 425, about 450, about 475, about 500,
about 525, about 550, about 575, about 600, about 625, about 650, or about 675
pg day/mL, and an area under the plasma concentration-time curve from time
zero to infinity (AUC.) of about 200, about 225, about 250, about 275, about
300,
about 325, about 350, about 375, about 400, about 425, about 450, about 475,
about 500, about 525, about 550, about 575, about 600, about 625, about 650,
about 675, about 700, about 725, about 750, or about 775 pg day/mL.
[0203] In some embodiments, a single SC administration of about 100 mg to a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from: an average T. of about 6 days, an average C. of about 13
pg/mL, an average area under the plasma concentration-time curve from time
zero to time of last measurable concentration (AUCiast) of about 421 pg
day/mL,
and an average area under the plasma concentration-time curve from time zero
to infinity (AUC.) of about 477 pg day/mL.
[0204] In some embodiments, a single SC administration of about 100 mg
achieves one or more pharmacokinetic characteristics chosen from a clearance
rate (CL/F) of about 100, about 125, about 150, about 175, about 200, about
225,
about 250, about 275, about 300, about 325, about 350, about 375, about 400,
about 425, or about 450 mL/day, an apparent volume of distribution (Vz/F) of
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
57
about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, about 8.0,
about
8.5, about 9.0, about 9.5, about 10, about 10.5, about 11, about 11.5 or about
12
L, and a serum half-life of about 15 days, about 16 days, about 17 days, about
18
days, about 19 days, about 20 days, about 21 days, about 22 days, about 23,
about 24 days, about 25 days, about 26 days, about 27 days, about 28 days,
about 29 days, about 30 days, about 31 days, about 32 days, about 33 days, or
about 34 days.
[0205] In some embodiments, a single SC administration of about 100 mg to a
population of subjects achieves one or more pharmacokinetic characteristics
chosen from an average clearance rate (CL/F) of about 275 mL/day, an apparent
volume of distribution (\//F) of about 8 L, and a serum half-life of about 25
days.
[0206] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 7-day (weekly) intervals to achieve a steady state,
and
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
Tmax ss of about 2, about 2.5, about 3, about 3.5, about 4, about 4.5, about
5,
about 5.5, about 6, or about 6.5 days, a Cmax ss of about 35, about 40, about
45,
about 50, about 55, about 60, about 65, about 70, about 75, about 80, about
85,
about 90, or about 95 pg/mL, an AUCT ss of about 225, about 250, about 275,
about 300, about 325, about 350, about 375, about 400, about 425, about 450,
about 475, about 500, about 525, about 550, about 575, about 600, about 625,
or
about 650 pg day/mL, and a Ctrough ss of about 35, about 40, about 45, about
50,
about 55, about 60, about 65, about 70, about 75, or about 80pg/mL is
achieved.
[0207] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 7-day (weekly) intervals to a population of subjects
to
achieve a steady state, and wherein one or more steady state pharmacokinetic
characteristics chosen from: an average Tmax ss of about 4 days, an average C.
ss of about 65 pg/mL, an average AUCT ss of about 443 pg day/mL, and a Ctrough
ss
of about 59 pg/mL is achieved.
[0208] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 7-day (weekly) intervals to achieve a steady state,
and
wherein one or more steady state pharmacokinetic characteristics chosen from:
a
clearance rate (CLss/F) of about 150, about 160, about 170, about 180, about
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
58
190, about 200, about 210, about 220, about 230, about 240, about 250, about
260, about 270, about 280, about 290, about 300, about 310, about 320, about
330, about 340, about 350, about 360, about 370, about 380, about 390, or
about
400 mL/day, an apparent volume of distribution (\//F) of about 7, about 7.5,
about 8, about 8.5, about 9, about 9.5, about 10, about 10.5, about 11, about
11.5, about 12, about 12.5, about 13, about 13.5, about 14, about 14.5, or
about
15 L, and a serum half-life of about 22 , about 23, about 24 of less, about
25,
about 26, about 27, about 28, about 29, about 30, about 31, about 32, about
33,
about 34, or about 35 days is achieved.
[0209] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 7-day (weekly) intervals to a population of subjects
to
achieve a steady state, and wherein one or more steady state pharmacokinetic
characteristics chosen from an average clearance rate (CLõ) of about 282
mL/day, an average apparent volume of distribution (\//F) of about 11 L, and
an
average serum half-life of about 28 days is achieved.
[0210] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 14-day (bi-weekly) intervals to achieve a steady
state,
and wherein one or more steady state pharmacokinetic characteristics chosen
from: a Tmax ss of about 2, about 2.5, about 3, about 3.5, about 4, about 4.5,
about
5, about 5.5, about 6, about 6.5, or about 7 days, a Cmax ss of about 30,
about 32,
about 34, about 36, about 38, about 40, about 42, about 44, about 46, about
48,
or about 50 pg/mL, an AUCT ss of about 420 , about 430, about 440, about 450,
about 460, about 470, about 480, about 490, about 500, about 510, about 520,
about 530, about 540, about 550, about 560, or about 570 pg day/mL, and a
Ctrough ss of about 20, about 21, about 22, about 23, about 24, about 25,
about 26,
about 27, about 28, about 29, about 30, about 31, about 32, about 33, about
34,
about 35, about 36, about 37, about 38, about 39, or about 40 pg/mL is
achieved.
[0211] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 14-day (bi-weekly) intervals to a population of
subjects
to achieve a steady state, and wherein one or more steady state
pharmacokinetic
characteristics chosen from: an average Tmax ss of about 4 days, an average C.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
59
ss of about 39 pg/mL, an average AUCT ss of about 495 pg day/mL, and a Ctrough
ss
of about 30 pg/mL is achieved.
[0212] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 14-day (bi-weekly) intervals to achieve a steady
state,
and wherein one or more steady state pharmacokinetic characteristics chosen
from: a clearance rate (CLss/F) of about 170, about 175, about 180, about 185,
about 190, about 195, about 200, about 205, about 210, about 215, about 220,
about 225, about 230, about 235, or about 240 mL/day, an apparent volume of
distribution (Vz/F) of about 6 of less, about 6.5, about 7, about 7.5, about
8, about
8.5, about 9, about 9.5, or about 10 L, and a serum half-life of about 18,
about 20,
about 22, about 24, about 26, about 28, about 30, about 32, about 34, about
36,
or about 38 days is achieved.
[0213] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 14-day (bi-weekly) intervals to a population of
subjects
to achieve a steady state, and wherein one or more steady state
pharmacokinetic
characteristics chosen from an average clearance rate (CL) of about 406
mL/day, an average apparent volume of distribution (Vz/F) of about 8 L, and an
average serum half-life of about 28 days is achieved.
[0214] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 30-day (monthly) intervals to achieve a steady
state,
and wherein one or more steady state pharmacokinetic characteristics chosen
from: a Tmax ss of about 3, about 3.5, about 4, about 4.5, about 5, about 5.5,
about
6, about 6.5, about 7, about 7.5, or about 8 days, a C max ss of about 14,
about 15,
about 16, about 17, about 18, about 19, about 20, about 21, about 22, about
23,
about 24, about 25, about 26, about 27, about 28, about 29, about 30, about
31,
about 32, about 33, or about 34 pg/mL, an AUCT ss of about 325, about 350,
about
375, about 400, about 425, about 450, about 475, about 500, about 525, about
550, about 575, about 600, about 625, or about 650 pg day/mL, and a Ctrough ss
of
about 6, about 6.5, about 7, about 7.5, about 8, about 8.5, about 9, about
9.5,
about 10, about 10.5, about 11, about 11.5, about 12, about 12.5, about 13,
about 13.5, about 14, about 14.5, or about 15 pg/mL is achieved.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
[0215] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 30-day (monthly) intervals to a population of
subjects
to achieve a steady state, and wherein one or more steady state
pharmacokinetic
characteristics chosen from: an average Tmax ss of about 6 days, an average C.
ss of about 49 pg/mL, an average AUCT ss of about 483 pg day/mL, and a Ctrough
ss
of about 11 pg/mL is achieved.
[0216] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 30-day (monthly) intervals to achieve a steady
state,
and wherein one or more steady state pharmacokinetic characteristics chosen
from: a clearance rate (CLss/F) of about 150, about 160, about 170, about 180,
about 190, about 200, about 210, about 220, about 230, about 240, about 250,
about 260, about 270, about 280, about 290, about 300, or about 310 mL/day, an
apparent volume of distribution (Vz/F) of about 5, about 6, about 7, about 8,
about
9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, or
about
17 L, and a serum half-life of about 18 , about 20, about 22, about 24, about
26,
about 28, about 30, about 32, about 34, about 36, about 38, about 40, about
42,
about 44, about 46, or about 48 days is achieved.
[0217] In some embodiments, a sufficient number of SC doses of about 100 mg
are administered at about 30-day (monthly) intervals to a population of
subjects
to achieve a steady state, and wherein one or more steady state
pharmacokinetic
characteristics chosen from an average clearance rate (CL) of about 227
mL/day, an average apparent volume of distribution (Vz/F) of about 11 L, and
an
average serum half-life of about 33 days is achieved.
Dosing to Achieve Desired Pharmacodynamic Characteristics
[0218] The term "pharmacodynamic characteristics" includes parameters
describing the biological effects of an administered drug, e.g., an anti-IFN-
alpha
antibody such as sifalimumab or an antigen-binding fragment thereof. One such
characteristic is the expression of specific genes, which can serve as PD
markers. In particular, certain genes when overexpressed in a patient
suffering
from an autoimmune disease relative to the genes' expression in a healthy
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
61
patient, or relative to the abundance of housekeeping genes, can serve as a
type
I IFN or IFN-alpha-inducible PD marker expression profile.
[0219] The group of genes included in the type I IFN or IFN-alpha-inducible
PD
marker expression profile of the patient are (a) IF127, IF144, IF144L, IFI6
and
RSAD2; or (b) IF144, IF144L, IFI6 and RSAD2; or (c) IF127, IF144L, IFI6 and
RSAD2; or (d) IF127, IF144, IFI6 and RSAD2; or (e) IF127, IF144, IF144L, and
RSAD2; or (f) IF127, IF144, IF144L, and IF16.
[0220] In a specific embodiment, the group of genes included in the type I
IFN or
IFN-alpha-inducible PD marker expression profile of the patient comprises
IF127,
IF144, IF144L, IFI6 and RSAD2. In another specific embodiment, the group of
genes included in the type I IFN or IFN-alpha-inducible PD marker expression
profile of the patient consists of IF127, IF144, IF144L, IFI6 and RSAD2. In a
further
specific embodiment, the group of genes included in the type I IFN or IFN-
alpha-
inducible PD marker expression profile of the patient comprises IF127, IF144,
IF144L, and RSAD2. In another specific embodiment, the group of genes
included in the type I IFN or IFN-alpha-inducible PD marker expression profile
of
the patient consists of IF127, IF144, IF144L, and RSAD2.
[0221] The IFN-alpha-inducible PD markers in an expression profile may
include
(a) IF127, IF144, IF144L, IFI6 and RSAD2; or (b) IF144, IF144L, IFI6 and
RSAD2;
or (c) IF127, IF144L, IFI6 and RSAD2; or (d) IF127, IF144, IFI6 and RSAD2; or
(e)
IF127, IF144, IF144L, and RSAD2; or (f) IF127, IF144, IF144L, and IF16.
[0222] The IFN-alpha-inducible PD markers in an expression profile may
consist
of (a) IF127, IF144, IF144L, IFI6 and RSAD2; or (b) IF144, IF144L, IFI6 and
RSAD2; or (c) IF127, IF144L, IFI6 and RSAD2; or (d) IF127, IF144, IFI6 and
RSAD2; or (e) IF127, IF144, IF144L, and RSAD2; or (f) IF127, IF144, IF144L,
and
IF16.
[0223] An alternative set of genes that may serve as PD markers includes
the 21
genes IF144, IF127, IF144L, DNAPTP6, LAMP3, LY6E, RSAD2, HERC5, IF16,
I5G15, 0A53, SIGLEC1, 0A52, USP18, RPT4, IFIT1, MX1, OAS1, EPST1,
PLSCR1, and IFRG28, as described in U.S. Patent Application 12/598, 526, file
May 5, 2008. See Table 24 following paragraph [0500].
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
62
[0224] The upregulation or downregulation of the type I IFN or IFN-alpha-
inducible PD markers in the patient's expression profile may be by any degree
relative to that of a sample from a control (which may be from a sample that
is not
disease tissue of the patient (e.g., non-lesional skin of a psoriasis patient)
or from
a healthy person not afflicted with the disease or disorder) or may be
relative to
that of genes from the patient whose expression is not changed by the disease
(so called "house keeping" genes.)
[0225] The degree upregulation or downregulation may be at least 10%, at
least
15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at
least
60%, at least 70%, at least 75%, at least 80%, at least 85, at least 90%, at
least
95%, at least 100%, at least 125%, at least 150%, or at least 200%, or at
least
300%, or at least 400%, or at least 500% or more that of the control or
control
sample.
[0226] Type I IFN or IFN-alpha-inducible PD marker expression profile may
be
calculated as the average fold increase in the expression or activity of the
set of
genes comprised by the PD marker. The Type I IFN or IFN-alpha-inducible PD
marker expression profile may also be calculated as the difference between the
mean Ct (cycle threshold) for the four target genes and the mean Ct of three
control genes.
[0227] The average fold increase in the expression or activity of the set
of genes
may be between at least about 2 and at least about 15, between at least about
2
and at least about 10, or between at least about 2 and at least about 5. The
average fold increase in the expression or activity of the set of genes may be
at
least about 2, at least about 2.5, at least about 3, at least about 3.5, at
least
about 4, at least about 4.5, at least about 5, at least about 5.5, at least
about 6, at
least about 6.5, at least about 7, at least about 8, at least about 9 or at
least
about 10.
[0228] The degree of increased expression permits the identification of a
fold
change cutoff for identifying signature positive and signature negative
patients
suffering from autoimmune diseases. In one embodiment, the cutoff is at least
about 2. In another embodiment, the cutoff is at least about 2.5. In another
embodiment, the cutoff is at least about 3. In another embodiment, the cutoff
is
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
63
at least about 3.5. In another embodiment, the cutoff is at least about 4. In
another embodiment, the cutoff is at least about 4.5. In another embodiment,
the
cutoff is chosen from at least 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3,
4.4, and 4.5.
In another embodiment the cutoff is between about 2 and about 8. In one
embodiment, the cutoff is the mean of the increased expression levels of at
least
four of IF127, IF144, IF144L, IFI6 and RSAD2. In another embodiment, the
cutoff
is the median of the increased expression levels of at least four of IF127,
IF144,
IF144L, IFI6 and RSAD2.
[0229] The degree of increased expression also permits the identification
of a
delta Ct cutoff for identifying signature positive and signature negative
patients
suffering from autoimmune diseases. In one embodiment, the cutoff is at least
about 7.6. In another embodiment, the cutoff is 7.56. The fold change cutoff
may
be used to determine an appropriate delta Ct cutoff (e.g., 1 < log2 of the
fold
change < 3 corresponds to delta Ct range of 8.65 to 6.56.). Thus, in another
embodiment, the delta Ct cutoff is between about 6.56 to about 8.56.
[0230] Furthermore, the patient may overexpress or have a tissue that
overexpresses a type I IFN subtype at least 10%, at least 15%, at least 20%,
at
least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least
70%, at
least 75%, at least 80%, at least 90%, at least 100%, at least 125%, at least
150%, or at least 200%, or at least 300%, or at least 400%, or at least 500%
that
of the control. The type I IFN subtype may be any one of IFNalpha 1,
IFNalpha2,
IFNalpha4, IFNalpha5, IFNalpha6, IFNalpha7, IFNalpha8, IFNalphal0,
IFNalphal4, IFNalphal7, IFNalpha21, IFNbeta, or IFNomega. The type I IFN
subtypes may include all of IFNalphal, IFNalpha2, IFNalpha8, and IFNalphal4.
[0231] The up-regulated expression or activity of any gene detected in a
sample,
by probes, or by probes in kits in an IFN-alpha-inducible PD marker expression
profile may be at least 1.2-fold, at least 1.25-fold, at least 1.3-fold, at
least 1.4-
fold, at least 1.5-fold, at least 2.0-fold, at least 2.25-fold, at least 2.5-
fold, at least
2.75-fold, at least 3.0-fold, at least 3.5-fold, at least 4.0-fold, at least
4.5-fold, at
least 5.0-fold, at least 6.0-fold, at least 7.0-fold, at least 8.0-fold, at
least 9.0-fold,
at least 10.0-fold, at least 15.0-fold, at least 20.0-fold, at least 25.0-
fold, or at
least 50.0-fold relative to baseline levels of control cells, e.g., cells of
healthy
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
64
volunteers or cells of control animals or cells not exposed to IFN in culture.
All
of the genes in the IFN-alpha-inducible PD marker expression profile may have
up-regulated expression or activity at the same fold increase. Alternatively,
the
genes in the PD marker expression profile may have varying levels of up-
regulated expression or activity.
Measuring Upregulation
[0232] Up- or down-regulation of gene expression or activity of IFN-alpha-
inducible PD markers may be determined by any means known in the art. For
example, up- or down-regulation of gene expression may be detected by
determining mRNA levels. mRNA expression may be determined by northern
blotting, slot blotting, quantitative reverse transcriptase polymerase chain
reaction, or gene chip hybridization techniques. See U.S. Pat. Nos. 5,744,305
and 5,143,854 for examples of making nucleic acid arrays for gene chip
hybridization techniques. See Establishing and functional characterization of
an
HEK-293 cell line expressing autofluorescently tagged 13 -actin (pEYFP-ACTIN)
and the neurokinin type 1 receptor (NK1-R) Hrovat, A; Zavec, AB; Pogacnik, A;
Frangez, R; Vrecl, M 2010 Cellular & Molecular Biology Letters 1, 55-69,
Expression profiles of proliferative and antiapoptotic genes in sporadic and
colitis-
related mouse colon cancer models Svec, J; Ergang, P; Mandys, V; Kment, M;
Pacha, J 2010 International Journal of Experimental Pathology 1, 44-53, and
Protein kinase inhibitors emodin and dichloro-ribofuranosylbenzimidazole
modulate the cellular accumulation and cytotoxicity of cisplatin in a schedule-
dependent manner Kurokawa, T; He, GA; Siddik, ZH 2010 Cancer
Chemotherapy and Pharmacology 3, 427-436, for examples of how to use the
TAQMANO method for measuring gene expression.
[0233] Primers that selectively bind to targets in polymerase chain
reactions
(PCR) can be chosen based on empirically determining primers that hybridize in
a PCR reaction and produce sufficient signal to detect the target over
background, or can be predicted using the melting temperature of the
primer:target duplex as described in Maniatis et al. Molecular Cloning, Second
Edition, Section 11.46. 1989. Similarly, probes for detecting PCR products in
a
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
TAQMANO or related method can be empirically chosen or predicted. Such
primers and probes (collectively "oligonucleotides") may be between 10 and 30
nucleotides or greater in length.
[0234] Up- or down-regulation of gene expression or activity of IFN-
alpha-
inducible PD markers may be determined by detecting protein levels. Methods
for detecting protein expression levels include immuno-based assays such as
enzyme-linked immunosorbant assays, western blotting, protein arrays, and
silver
staining.
[0235] An IFN-alpha-inducible PD marker expression profile may comprise
a
profile of protein activity. Up- or down-regulation of gene expression or
activity of
IFN-alpha-inducible PD markers may be determined by detecting activity of
proteins including, but not limited to, detectable phosphorylation activity,
de-
phosphorylation activity, or cleavage activity.
Furthermore, up- or down-
regulation of gene expression or activity of IFN-alpha-inducible PD markers
may
be determined by detecting any combination of these gene expression levels or
activities.
Neutralization of the type I IFN or IFN-alpha-inducible profile in patients
[0236] Treatment with the anti-IFN-alpha antibody or antigen-binding
fragment
thereof neutralizes the type I IFN or IFN-alpha-inducible profile. Treatment
with
the anti-IFN-alpha antibody or antigen-binding fragment thereof results in a
decrease in one or more symptoms of the type I IFN or an IFN-alpha-mediated
disease or disorder. Treatment with the anti-IFN-alpha antibody or antigen-
binding fragment thereof results in fewer flare-ups related to the type I IFN
or an
IFN-alpha-mediated disease or disorder. Treatment with the anti-IFN-alpha
antibody or antigen-binding fragment thereof results in improved prognosis for
the
patient having the type I IFN or an IFN-alpha-mediated disease or disorder.
Treatment with the anti-IFN-alpha antibody or antigen-binding fragment thereof
results in a higher quality of life for the patient. Treatment with the anti-
IFN-alpha
antibody or antigen-binding fragment thereof alleviates the need to co-
administer
second agents (e.g., steroids) or may lessen the dosage of administration of
the
second agent to the patient. Treatment with the anti-IFN-alpha antibody or
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
66
antigen-binding fragment thereof reduces the number of hospitalizations of the
patient that are related to the type I IFN or an IFN-alpha-mediated disease or
disorder.
[0237] The anti-IFN-alpha antibody or antigen-binding fragment thereof
neutralizes a type I IFN or IFN-alpha-inducible profile. Neutralization of the
type I
IFN or IFN-alpha-inducible profile may be a reduction in at least one, at
least two,
at least three, at least four genes. Neutralization of the type I IFN or IFN-
alpha-
inducible profile is a reduction of at least 2%, at least 3%, at least 4%, at
least
5%, at least 7%, at least 8%, at least 10%, at least 15%, at least 25%, at
least
30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at
least
70%, at least 75%, at least 80%, or at least 90% of any of the at least one,
at
least two, at least three, at least four genes up-regulated in the type I IFN
or IFN-
alpha-inducible profile.
[0238] Alternatively, neutralization of the type I IFN or IFN-alpha-
inducible profile
refers to a reduction of expression of up-regulated type I IFN or IFN-alpha-
inducible genes that is within at most 50%, at most 45%, at most 40%, at most
35%, at most 30%, at most 25%, at most 20%, at most 15%, at most 10%, at
most 5%, at most 4%, at most 3%, at most 2%, or at most 1% of expression
levels of those type I IFN or IFN-alpha-inducible genes in a control sample.
The
anti-IFN-alpha antibody or fragment thereof may neutralize the type I IFN or
IFN-
alpha profile at doses of 0.3 to 30 mg/kg, 0.3 to 10 mg/kg, 0.3 to 3 mg/kg,
0.3 to 1
mg/kg, 1 to 30 mg/kg, 3 to 30 mg/kg, 5 to 30 mg/kg, 10 to 30 mg/kg, 1 to 10
mg/kg, 3 to 10 mg/kg, or 1 to 5 mg/kg. In a specific embodiment, the type I
IFN
or IFN-alpha profile is neutralized by about 40% when sifalimumab is
administered with weekly, subcutaneous dosing of 100 mg.
[0239] All of the references cited above, as well as all references cited
herein, are
incorporated herein by reference in their entireties.
[0240] The following examples are offered by way of illustration and not by
way of
limitation.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
67
EXAM PLES
Example 1
Intravenous Pharmacokinetics and Immunogenicity of Body Weight-Based
Dosing Regimes of Sifalimumab Administered Intravenously to SLE
Patients
[0241] The PK and immunogenicity (IM) of intravenous (IV) weight-based
doses
of sifalimumab administered every 14 days was studied in a Phase lb study in
adult patients with moderate to severe SLE to evaluate the safety profile of
sifalimumab.
1. Methods
1.1 Study Design
[0242] This was a multicenter, randomized, double-blind, placebo-
controlled,
dose-escalation study with 4 arms for a total of 14 doses in 161 patients with
moderate to severe SLE. 161 patients were randomized in a 3:1 ratio to receive
sifalimumab or placebo. Sifalimumab was administered every 14 days as a 60-
minute IV infusion. Four IV sifalimumab dosages were used: 0.3, 1.0, 3.0, and
10
mg/kg.
[0243] Criteria for inclusion of subjects receiving the antibody or antigen
binding
fragment thereof intravenously are summarized in clinical trial identifier
NCT00482989, accessible via the U.S. National Institutes of Health
clinicaltrials.gov database. Thus, inclusion criteria comprise: Male or female
adults were between about 18 and about 95 years of age at the time of the
first
dose of study drug; subjects met at least 4 of the 11 revised American College
of
Rheumatology classification criteria for SLE (Appendix A, ACR,1999); subjects
had positive anti-nuclear antibody (ANA) test at 1:80 serum dilution in the
past
or at screening; and subjects had at least one system with a score of A or two
systems with a score of B on the BILAG index at screening, or had a SELENA-
SLEDAI score 6.
[0244] Exclusion criteria comprised having received MEDI-545 within 120
days
prior to screening or have either detectable levels of MEDI-545 or anti-MEDI-
545
antibodies (positive at > 1:10 serum dilution) in serum at screening; history
of
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
68
allergy or reaction to any component of the study drug formulation; have
received
prednisone > 20 mg/day (or an equivalent dose of another oral
corticosteroid)within 14 days before randomization/entry; having received the
following dosages of medications within 28 days before randomization/entry:
hydroxychloroquine > 600 mg/day, mycophenolate mofetil > 3 g/day,
methotrexate > 25 mg/week, azathioprine > 3 mg/kg/day, or any dose of
cyclophosphamide, cyclosporine, or thalidomide; having received leflunomide
>20 mg/day in the 6 months prior to Study Day 0; having received fluctuating
doses of antimalarials, mycophenolate mofetil, methotrexate, leflunomide, or
azathioprine within 28 days before randomization/entry or fluctuating doses of
NSAIDs or oral corticosteroids within 14 days before randomization/entry;
treatment with any investigational drug therapy within 28 days before
randomization/entry into the study, B cell-depleting therapies within 12
months
before randomization/entry, or biologic therapies within 30 days or 5 half-
lives of
the biologic agent, whichever was longer, before randomization/entry into the
study; evidence of clinically significant active infection, including ongoing,
chronic
infection, within 28 days before randomization/entry; history of severe viral
infection as judged by the investigators, including severe infections of
either
cytomegalovirus or the herpes family such as disseminated herpes, herpes
encephalitis, ophthalmic herpes; herpes zoster infection within 3 months
before
randomization/entry; evidence of infection with hepatitis B or C virus, or HIV-
1 or
HIV-2, or active infection with hepatitis A, as determined by results of
testing at
screening; vaccination with live attenuated viruses within 28 days before
randomization/entry; pregnancy (women, unless surgically sterile or at least 2
years post-menopausal, had a negative serum pregnancy test within 28 days
before receiving the study drug and a negative urine pregnancy test on Study
Day 0 before receiving the study drug); breastfeeding or lactating women;
history
of primary immunodeficiency; history of alcohol or drug abuse < 1 year prior
to
randomization/entry; history of cancer (except basal cell carcinoma or in situ
carcinoma of the cervix treated with apparent success with curative therapy >
1
year prior to randomization/entry); history of active TB infection; history of
latent
TB infection or newly positive TB skin test (reaction defined as 10 mm
in
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
69
diameter if not on systemic immunosuppressive medication or 5 mm
if on
systemic immunosuppressive medication) without completion of an appropriate
course of treatment or with ongoing prophylactic therapy; elective surgery
planned from the time of screening through Study Day 196; at screening blood
tests (within 28 days before randomization/entry), any of the following (i)
AST > 2
x upper limit of normal range (ULN), unless caused by SLE, as determined by
the
investigator, (ii) ALT > 2 x ULN, unless caused by SLE, as determined by the
investigator, (iii) Creatinine > 4.0 mg/dL, (iv) Neutrophils "1,500/ pL (< 1.5
x
109/L)," (v) Platelet count "Platelet count < 50,000/ pL (< 50 x 109/L)";
history of
any disease, evidence of any current disease (other than SLE), any finding
upon
physical examination, or any laboratory abnormality that, in the opinion of
the
investigator or medical monitor, may have compromised the safety of the
patient
in the study or confound the analysis of the study.
[0245] The inclusion and exclusion criteria listed above are not
intended to limit
the scope of the present disclosure. A person skilled in the art would
understand
that other inclusion and/or exclusion parameters may be used to generate a
subject population such that the choice of subject does not compromise the
safety of the subject or confounds the analysis of the study.
[0246] Trough serum samples were collected for PK concentrations and IM
titers
at multiple time points. Samples were analyzed for PK using a validated enzyme
linked immunosorbent assays (ELISA) and for IM using a validated bridging
electrochemiluminescent assay (ECL).
1.2. Measurement of Anti-sifalimumab Antibodies in Human Serum Samples
Using a Validated Colorimetric Bridging ELISA Method (pre-study screen
samples)
[0247] As part of the pre-study evaluation of subjects for inclusion in
the study,
serum was evaluated for anti-sifalimumab antibody responses. Serum samples
were measured for the presence of anti-sifalimumab antibodies using a
colorimetric bridging ELISA. Serum samples were diluted 1:10 and were added to
a microtiter plate coated with sifalimumab. Following a wash step,
biotinylated-
sifalimumab was added to bind the captured anti-sifalimumab antibodies. Plates
were washed and streptavidin conjugated with horseradish peroxidase was
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
added, followed by tetramethylbenzidine substrate for detection of bridged
complexes. Plates were measured at a 450 nm wavelength using a Molecular
Devices SpectraMax microplate reader and the results were analyzed using
SOFTmax PRO software. The color intensity of the reaction was proportional to
the amount of anti-sifalimumab antibodies present in the sample.
[0248] The assay employed three positive controls ranging from 15 to 1500
ng/mL and one negative spiked control (0.75 ng/mL) that were prepared by
adding goat anti-sifalimumab anti-idiotypic antibody into pooled normal human
serum. The negative/positive cutoff value for samples was determined for each
assay plate and was calculated by multiplying the mean value of six unique
human serum samples measured on the plate by 1.5. Samples with responses
below the plate cutoff value were classified as negative and were assigned a
titer
value of < 10, (less than the reciprocal of the minimum required sample
dilution).
Samples with responses greater than or equal to the plate cutoff value were
classified as positive and were subsequently tested for titer. Titers were
performed by serially diluting samples with pooled normal human serum matrix
and were reported as the reciprocal of the highest 1:2 dilution (over the 1:10
minimum required sample dilution) that measured positive in the assay, before
returning a negative response.
[0249] Method validation met the acceptance criteria for accuracy and
precision
of classification and titer, intermediate precision, repeatability,
robustness,
linearity of dilution, specificity of detection, analyte stability in human
serum and
selectivity in normal human and SLE patient serum samples. The estimated
concentration of the cutoff of the assay was determined to be 4.5 ng/mL using
the
polyclonal goat anti-sifalimumab antibody surrogate control. Concentrations of
500 ng/mL anti-sifalimumab antibody (but not 100 ng/mL) were detectable in
serum samples containing 100 ng/mL sifalimumab drug.
1.3. Measurement of sifalimumab in Human Serum Using a Validated
Colorimetric ELISA Method
[0250] Sifalimumab was measured in human serum samples using a colorimetric
ELISA method that was validated for human serum. In the assay, microtiter
plates were coated with 0.5pg/mL goat anti-sifalimumab idiotype antibody,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
71
blocked with casein buffer and washed. Calibration standards (0.3 to 160
pg/mL)
and control samples were prepared by diluting sifalimumab reference standard
into human serum Standards, controls and unknown samples were diluted
1:1000 in assay buffer that contained 0.5% casein and 5% goat serum and were
added to the plate at 50 pL/well.
[0251] Following incubation, plates were washed to remove unbound material
and goat anti-human IgG conjugated with horseradish peroxidase was added for
binding of captured sifalimumab. Plates were washed and pre-warmed 2,2'-azino-
bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) peroxidase substrate was
added. The reaction was stopped by addition of 1% sodium dodecyl sulfate
(SDS) stop solution.
[0252] Plates were measured at a 405 nm wavelength using a Molecular
Devices
SpectraMax microplate reader and the results were analyzed using
SOFTmax PRO software. The color intensity of the reaction was proportional to
the amount of sifalimumab present in the sample. Concentrations of sifalimumab
in quality controls and unknown samples were interpolated from standard curves
using a 4-parameter logistic fit. The assay lower limit of quantitation (LLOQ)
was
determined to be 1.25 pg/mL and the upper limit of quantitation (ULOQ) was
determined to be 40 pg/mL.
[0253] Method validation demonstrated acceptable accuracy, repeatability,
intermediate precision, selectivity (evaluated in serum samples from normal
humans and humans with SLE), specificity, dilutional linearity, robustness,
and
stability of sifalimumab in human serum. Quantitation of sifalimumab at levels
of
3.0 and 36.0 pg/mL was not affected in the presence of 2 ng/mL interferon
alpha
target.
1.4. Measurement of Anti-sifalimumab Antibodies in Human Serum Samples
Using a Validated, Sensitive ECL Method
[0254] An ECL, solution-phase, bridging immunoassay that employs Meso Scale
Discovery (MSD) technology was developed and validated for the detection,
confirmation and titration of anti-sifalimumab antibodies in human serum. In
the
method, biotinylated sifalimumab and ruthenylated sifalimumab were incubated
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
72
overnight with human serum samples. Because of the bivalent nature of
antibodies, a portion of anti-sifalimumab antibodies (ADA) present in the
sample
bound to both conjugated forms of sifalimumab at the same time. Subsequently,
samples were incubated on a streptavidin-coated MSD plate for capture of the
ADA-bridged complexes.
[0255] The plate was then washed to remove unbound materials, read buffer
was added and the plate was placed on the MSD SectorTM Imager for generation
and measurement of ECL response. Application of an electrical current to the
electrode-containing plate caused the ruthenium chelate conjugated to
sifalimumab to emit light in the presence of the tripropylamine-containing
read
buffer. Samples that contained ADA bound to both biotin and ruthenium
conjugated forms of sifalimumab generated ECL signals. The signal intensity,
measured by the MSD Sector Imager, was proportional to the amount of anti-
sifalimumab antibodies present in the sample.
[0256] The method employed pooled normal human serum as the negative
control and pooled normal human serum spiked with a goat anti-sifalimumab
idiotype antibody (surrogate control) at two levels above detection (3.0 and
1000
ng/mL) as positive controls. The presence of anti- sifalimumab antibodies was
determined relative to a cutoff ECL value that was calculated for each plate
as
the mean response of 6-8 wells of the negative control multiplied by a 1.18
cut
point factor. The 1.18 cut point factor was established during validation from
200
measurements of serum samples obtained from 50 normal individuals and was
statistically determined to provide a 5% false positive rate. Samples that
measured at or above the cut point ECL value were considered as potential
positives for anti-sifalimumab antibodies and were retested in a confirmatory
(specificity) assay, both in the absence and presence of excess (300 g/mL)
sifalimumab.
[0257] The confirmatory cut point was established during method validation
using
the percent inhibition measurements of the above-mentioned samples tested
both in the absence and presence of excess (300 g/mL) sifalimumab. The
confirmatory cut point was statistically determined to provide a 0.1% false
positive rate and was determined to be 27.0%. The screening cut point factor
and
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
73
confirmatory cut point for SLE serum samples were evaluated using the
measured values of serum samples from fifty individual SLE patients and were
calculated to be 1.23% and 37.1%, respectively. Study samples were considered
positive for anti-sifalimumab antibodies if the percent inhibition of response
in the
presence of excess sifalimumab was greater than or equal to 27%, the more
conservative confirmatory cut point. Confirmed positive samples were then
measured in a titer assay.
[0258] Titers were performed by serially diluting samples with negative
control
serum and were reported as the reciprocal of the highest 1:2 dilution (over
the
1:10 minimum required sample dilution) that measured positive in the assay,
before returning a negative response. Titer values for negative samples were
reported as <10. Method validation met the acceptance criteria for accuracy
and
precision of classification and titer, intermediate precision, repeatability,
robustness, linearity of dilution, specificity of detection and analyte
stability in
human serum. Assay sensitivity was estimated using the polyclonal goat anti-
sifalimumab antibody surrogate control as well as a monoclonal anti-
sifalimumab
antibody of known affinity (Kd=1.30 nM). The approximate concentration of anti-
sifalimumab antibody at the cut point was 0.2 ng/mL for the polyclonal
antibody
and 4.7 ng/mL for the monoclonal antibody. Monoclonal anti-sifalimumab
antibody levels of 250-500 ng/mL and polyclonal anti-sifalimumab antibody
levels
of 500 ng/mL were detectable in serum containing 100 pg/mL of sifalimumab.
(provide temperature at which experiments were conducted).
1.5 Data Analysis
[0259] Pharmacokinetic concentrations were analyzed using non-compartmental
methods in WinNonlin (version 5.2.1 Pharsight, Cary, NC).
2. Results
2.1. Pharmacokinetics
[0260] Serum sifalimumab PK results following the first dose and after the
last
dose on Day 182 are summarized in TABLE 2 and TABLE 3, respectively. After
the first dose, sifalimumab peak plasma concentration (Cõx), area under the
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
74
plasma concentration-time curve from time zero to end of dosing interval
(AUCT)
and trough concentration (Ctrough) 1 increased dose-proportionally (TABLE 2).
Cmax
values ranged from 10.90 g/mL for the 0.3 mg/kg dosage to 229.74 g/mL for
the 10 mg/kg dosage. AUCT values ranged from 79.39 g.day/mL for the 0.3
mg/kg dosage to 1,610 g.day/mL for the 10 mg/kg dosage. Ctrough values ranged
from 2.75 g/mL for the 0.3 mg/kg dosage to 51.52 g/mL for the 10 mg/kg
dosage.
TABLE 2
Serum Sifalimumab PK Parameters Following the First Dose
Parameter 0.3 mg/kg IV 1 mg/kg IV 3 mg/kg IV
10mg/kg IV
T (day) 0.074 (56.5) 0.079 (53.7) 0.088
(51.6) 0.092 (45.2)
max
C (pg/mL) 10.90 (37.4) 31.89 (35.2) 103.34
(37.9) 229.74 (38.6)
max
AUCT (pg=day/mL) 79.39 (31.5) 220.9 (30.9) 739.4
(36.6) 1610 (39.2)
C (pg/mL) 2.75 (30.5) 8.01 (52.2) 23.43
(47.5) 51.52 (46.9)
trough
Data presented as mean (%CV). the %Cv is calculated as the Standard
deviation/mean*100 for the
evaluable subjects.
[0261] Steady state trough concentrations at each one of the tested doses
was
achieved in 3 months (FIG. 1, TABLE 3). Represented in the graph in FIG. 1 is
the mean and standard deviation value for the nominal time point (scheduled
blood draw) of available patients. If the value was below the limit of
quantification,
the value was treated as missing. Plotted values appear in TABLE 9.
[0262] Steady state peak plasma concentration (Cmax ss) values ranged from
17.74 g/mL for the 0.3 mg/kg dosage to 441.79 g/mL for the 10 mg/kg dosage.
Steady state area under the plasma concentration-time curve within a dosing
interval (AUCT SS) values ranged from 143.2 g.day/mL for the 0.3 mg/kg dosage
to 3,403 g.day/mL for the 10 mg/kg dosage. Steady state trough concentration
(Ctrough õ) values ranged from 7.89 g/mL for the 0.3 mg/kg dosage to 183.97
g/mL for the 10 mg/kg dosage.
[0263] After the last dose on Day 182, the mean steady state clearance
(CLss)
ranged from 185 to 238 mL/day, mean terminal half-life ranged from 20 to
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
29 days across the dosing groups, and steady state volume of distribution
(Vss)
ranged from 5 to 6 L across the dosing groups (TABLE 3).
TABLE 3
Serum Sifalimumab PK Parameters After Last Dose on Day 182
Parameter 0.3 mg/kg IV 1 mg/kg IV 3 mg/kg IV 10 mg/kg IV
T (day) 0.17 (252.0) 0.07 (59.6) 0.13
(152.2) 0.23 (256.3)
max ss
C (pg/mL) 17.74
(38.7) 48.08 (38.9) 153.07 (51.3) 441.79 (34.7)
max ss
AUCT SS (pg=day/mL) 143.2 (37.6) 401.8 (47.1) 1188 (55.1) 3403 (25.4)
(pg/mL) 7.89 (36.6) 19.58
(52.2) 50.06 (47.8) 183.97 (49.4)
ctrough ss
Half-life (day) 29.1 (47.4) 23.4 (34.9) 19.9 (29.8)
21.9 (30.7)
CL (mL/day) 185 (46.3) 233 (49.4) 220 (38.3) 238
(34.1)
ss
Vss (L) 6.30 (40.0) 6.35 (45.2) 5.02 (40.1)
5.58 (29.0)
Data presented as mean (CV%).
2.2. Immunogenicity
[0264] Twenty-seven out of 121 patients (22.3 %) receiving
investigational
product and one out of 40 patients (2.5%) receiving placebo tested positive
for
presence of anti-sifalimumab antibodies. Three out of 26 patients receiving
0.3
mg/kg doses of sifalimubab (11% incidence) (FIG. 2A), seven out of 25 patients
receiving 1 mg/kg doses (28% incidence) (FIG. 2B), seven out of 27 patients
receiving 3 mg/kg doses (25.9% incidence) (FIG. 2C), and ten out of 43
patients
receiving 10 mg/kg doses (23.3% incidence) (FIG. 2D) tested positive for the
presence of anti-sifalimumab antibodies. Positive titer values ranged from 10
to
1280, 17 subjects at or below 80, indicating low titers of anti-sifalimumab
antibody (FIG. 3).
[0265] The presence of anti-sifalimumab antibodies did not have an
impact
on sifalimumab clearance (FIG. 4). Accordingly, sifalimumab had an acceptable
safety and tolerability profile at the doses tested.
3. Conclusion
[0266] Sifalimumab PK was linear and dose-proportional following
intravenous administration over the dose range of 0.3 mg/kg to 10 mg/kg. Serum
clearance, volume of distribution and terminal half-life of sifalimumab were
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
76
representative of a monoclonal antibody without a significant antigen sink.
The
overall incidence of anti-sifalimumab antibodies was 22% with positive titers
ranging from 10 to 1280. The presence of anti-sifalimumab antibody did not
have
an impact on the pharmacokinetics of sifalimumab.
Example 2
Population Pharmacokinetics of Sifalimumab in SLE Patients
[0267] The
primary objectives of this analysis were to (a) model the
population pharmacokinetics (PK) of sifalimumab; (b) to identify and
quantitate
the impact of patient/disease characteristics on PK variability; and (c) to
evaluate
fixed versus body weight based dosing regimens.
[0268] Dosing based on fixed-body weight is uncommon, as most biologics
are
dosed based on body weight. But for an anti-Her2 antibody, the PK data
suggested fixed dosing might be possible. See, e.g., U.S. Patent No.
7,449,184.
Whether the PK data would support fixed dosing for sifalimumab was uncertain
because the body weight of the clinical trial subjects spanned 43 kg to 120
kg.
Moreover, unlike the Her2 antibody discussed in U.S. Patent No. 7,449,184,
sifalimumab binds multiple targets whose expression may vary among patients,
and thus has a potentially more complex PK profile
1. Population Pharmacokinetic Analysis Methodology
[0269]
Sifalimumab serum concentration-time data were collected from the
study described in Example 1. Sifalimumab serum concentrations were
determined with the validated colorimetric ELISA described in Example 1. A non-
linear mixed-effect modeling approach was used to analyze sifalimumab
pharmacokinetic data. The population pharmacokinetic modeling was performed
using NONMEM Version VII software (Globomax LLC, Ellicott City, MD, USA), G-
Fortran (http://gcc.gnu.org/fortran/) and Perl-speaks-NONMEM (PSN)
(http://psn.sourceforge.net/). Data management and graphical analyses were
performed using S-plus 8.1 (TIBCO Spotfire, Somerville, MA, USA), Xpose 4.0
(University of Uppsala, Uppsala, Sweden) and R 2.7.1 (http://cran.r-
project.org/)
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
77
software. The various steps involved in the modeling processes are described
below.
[0270] A series of structural models were evaluated for sifalimumab based
on
Akaike information criteria (AIC) value, objective function values, precision,
plausibility of parameter estimates, and goodness-of-fit plots. The between-
subject variability in pharmacokinetic parameters was assumed to follow a log-
normal distribution and was modeled using exponential functions. The residual
variability was evaluated using homoscedastic (additive), heteroscedastic
(proportional), or combined proportional and additive models. The precision of
the
population estimates was evaluated based on percent relative standard errors
(RSEs).
[0271] After the structural model was identified, covariate model-building
was
carried out to assess the effect of patient/disease characteristics on the
pharmacokinetic parameters. Various patient/disease characteristics including
age, gender, ethnicity, region, body weight (WT), baseline steroid use
(BSTEROID), baseline systemic lupus erythematosus disease activity Index
(BSLEDAI) score, baseline gene signature from 21 genes (BGENE21) and
baseline gene signature from 4 genes (BGENE4) were evaluated.
[0272] A preliminary assessment of covariate influence was conducted using
generalized additive modeling (GAM) approach as implemented in Xpose
(Jonsson et al, 1999). Based on GAM results and mechanistic understanding of
sifalimumab disposition, the relevant covariates for each parameter were
further
tested using NONMEM for their significance. The model building was carried out
using step-wise forward addition (p<0.05) (8,0FV > 3.84), approach followed by
backward elimination (p<0.01) (8,0FV > 6.63) process. The covariates were
included in the final model if p-value <0.01 (8,0FV > 6.63), provided the
covariates were reasonable based on the pharmacology of sifalimumab.
[0273] Improvement in the model at each step was assessed using following
criteria: (a) reduction in objective function value; (b) improvement in
agreement
between the observed and population/individual predicted serum concentrations;
(c) reduction in between and within-subject variability; (d) reduction in the
range
of weighted residuals; (e) uniformity of the distribution of weighted
residuals
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
78
versus the predicted concentrations around the line of identity; and (f)
improvement in parameter precision. All models were run using the first order
conditional estimation (FOCE) with interaction method.
[0274] The relationship between continuous covariates and pharmacokinetic
parameters were modeled using non-linear power functions [equation (1)] with
the covariate normalized to the population median for the data set. The
categorical covariates were modeled using fractional change functions
[equation
(2)].
r COV `82
P = Oix (1)
Median)
P = Oix (1+ 92 X CO V) (2)
[0275] Where the es are the parameters to be estimated and 01 represents
the
typical value of pharmacokinetic parameter (P) in an individual with the
median
value for the covariate. 02 represents the coefficient for particular
covariate
effect.
[0276] The performance of the final population pharmacokinetic model was
evaluated using visual predictive check (VPC), a technique whereby model
appropriateness is tested by means of comparing prediction intervals (Pis) of
the
observed data to simulation data using final model.
[0277] The impact of body-weight based and fixed dosing of sifalimumab was
evaluated by comparing predicted steady state serum concentrations (Cõ) and
variability using population model. A population of 1000 SLE subjects were
simulated using covariates (demographic, BGENE21, BSTEROID) distribution
form study MI-CP152. The population model was also utilized for predicting
pharmacokinetic exposure following various fixed intravenous doses of
sifalimumab to support phase II dosing.
2. Results
2.1 Data
[0278] A total of 120 patients provided evaluable PK data with a total of
2,370
serum concentrations (average of 20 samples per patient). One subject from the
mg/kg cohort was excluded from the analysis due to very low observed serum
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
79
concentrations compared to the average concentrations in the 10 mg/kg cohort.
TABLE 4 lists the summary of patient characteristics included in the
pharmacokinetic database. A total of 8 subjects (6.67%) did not have baseline
gene signature (4 genes) information available; hence a population median
value
was imputed for these subjects.
2.2 Population Pharmacokinetic Modeling
[0279] A 2-compartment model was used to fit sifalimumab concentration-time
data. The model was parameterized using clearance (CL), central volume of
distribution (Vc), peripheral volume of distribution (Vp) and the inter-
compartmental clearance (Q). The multiplicative covariate modeling approach
was used to study the influence of various covariates including age, gender,
ethnicity, region, WT, BSTEROID, BSLEDAI, BGENE21 and BGENE4.
[0280] The final model functions for typical value of CL, Vc, Vp and Q are
presented as follows (equations 3-6).
BGENE21`96 r Doset97 i
CL = al x ¨ x _______________________ x _________________________ x 0_ +
0,x BSTEROID) (3)
75 i 32 i 1 i
WT 89
V c= 02X
75 i
, WTt9"
V p= 03X
75 i
Q = 04 (6)
[0281] Where el is the typical CL of a standard subject with WT=75 kg,
BGENE21=32, Dose=1 mg/kg and BSTEROID=0. 02 and 03 represent the Vc and
Vp of a standard subject with WT=75 kg, respectively. 05 to 010 are the
exponents
of covariate effect on respective pharmacokinetic parameters.
[0282] The final population pharmacokinetic parameters are presented in
TABLE
5. The estimated values of CL, Vc, Vp and Q for a standard subject were about
176 mL/day, 2.9 L, 2.12 L and 171 mL/day, respectively. The estimates of
between-subject variability (CVs) associated with CL, Vc, Vp and Q were 28%,
31%, 58% and 71%, respectively. All of the pharmacokinetic parameters were
estimated with good precision, as reflected by RSEs. The performance of the
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
final model fit is represented by goodness of fit plots as shown in FIG. 5.
These
figures, FIG.5 panels (a) and (b), show good agreement between observed and
model predicted (population/individual predicted) sifalimumab serum
concentrations, as all the points are close to the line of identity and the
fitted
spline curve almost overlap the line of identity. The plots of weighted
residual
versus the population predicted concentrations, FIG. 5 panel (c), or time,
FIG. 5
panel (d), do not show any obvious pattern. VPC results demonstrated good
predictability of the final population PK model as shown in FIG. 6.
[0283] Based on the covariate relationships, higher sifalimumab CL was
estimated for subjects with higher baseline type I IFN gene signature (21
genes),
body weight, sifalimumab dose and steroid use. Both Vc and Vp also increased
with increase in baseline body weight. Although the above mentioned covariates
were identified as statistically significant covariates, they did not
substantially
explain inter-individual variability (<7%) in CL, Vc and Vp. This means that a
decrease or increase in concentrations in individual patients in a dose group
is
not governed by body weight since a minimal magnitude of the overall
variability
in concentrations is explained by body weight. Hence no dosing modifications
are
necessary to administer sifalimumab.
2.2.1 Comparison of Fixed (mg) Versus Body weight Based (mg/kg) Dosing
[0284] The impact of fixed versus body weight based dosing was
evaluated by
comparing 200 mg (fixed) with 3 mg/kg (body weight based) every 14-days
dosing of sifalimumab in a simulated SLE population of 1000 subjects. The body
weight distribution (43 kg -120 kg) from MI-CP152 was used for the
simulations.
The final population PK model was used to predict 5th, median and 95th
percentile concentration-time profiles. Simulation results demonstrate that
both
fixed and body weight based dosing regimens yield similar median steady state
concentrations (Css) and variability as shown in FIG. 7.
2.2.2 Predicted Serum Concentrations
[0285] The final population PK model was used to predict PK profiles
following
200, 600, and 1200 mg monthly (with an additional dose at Day 14) dose of
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
81
sifalimumab in a simulated SLE population of 1000 subjects. The predicted PK
profiles (median, 5th and 95th percentiles) are shown in FIG 8. The expected
steady state PK parameters following 200, 600, and 1200 mg monthly (with an
additional dose at Day 14) dose of sifalimumab are presented in TABLE 6.
Predicted PK exposure in 40 kg patient and 120 kg patients, at 5th and 95th
percentile, appear in TABLE 11.
TABLE 4
Patient demographic characteristics and pharmacodynamic biomarkers
Categorical Variables
No. of Categories Percentag
Patients e (%)
(N)
Treatment 26 0.3 21.7
Cohort
(mg/kg)
25 1 20.8
27 3 22.5
42 10 35
Sex 6 1 (Male) 5
114 2 (Female) 95
Region 85 1 (North America) 70.8
35 2 (South America) 29.2
BSTEROID 32 0 (no) 26.7
88 1 (yes) 73.3
Continuous Variables
No. of Mean SD Median Range
Patients (N)
WT (kg) 120 76 19 73 43.1-120
Age (yr) 120 42 11 43 18-71
BSLEDAI 120 11 5 10 2-34
BGENE21 120 32 24 33 0.63-87
BGENE4 120 81 87 56 0.26-337
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
82
BSTEROID = Baseline steroid use; WT = Baseline body weight; BSLEDAI = Baseline
SLEDAI
Scores; BGENE21 = Baseline gene signature for 21 genes; BGENE4 = Baseline gene
signature
for 4 genes;
TABLE 5
Population pharmacokinetic parameters of sifalimumab from the final model
Parameter (unit) Typical value Between-subject
(RSE, %) Variability, CV% (RSE, %)
CL, std {el} (L/day) 0.176 (7) 28 (13)
Vc, sta {02} (L) 2.90(3) 31 (19)
Vp, sta 1031 (L) 2.12(6) 58(35)
Q (L/day) 0.171 (12) 71 (33)
Exponent: WT on CL 051 0.481 (6)
Exponent: BGENE21 on CL 0.0558 (33)
Exponent: Dose on CL {07} 0.0542 (17)
Exponent: BSTEROID on CL 0.195 (38)
Exponent: WT on Vp {09} 0.489 (17)
Exponent: WT on Vp 0.646 (16)
Correlation between CL and Vp 0.557 (21)
Correlation between Vc and Vp 0.131 (8)
Residual error (CV, %) 27.5 (8)
CL = Linear clearance; Vp = Central volume of distribution; Vp = Peripheral
volume of
distribution; Q = Inter-compartmental clearance; BGENE21 = Baseline gene
signature for 21
genes; BSTEROID = Baseline steroid use (0=no and 1=yes); WT = Baseline body
weight; CV =
Coefficient of variance.
CL, std = Clearance of a standard subject with WT=75 kg, BGENE21=32, Dose=1
mg/kg and
BSTEROID=0; Vc, std = Central volume of distribution for a standard subject
with WT=75 kg; Vp,
std = Peripheral volume of distribution for a standard subject with WT=75 kg.
TABLE 6
Sifalimumab Predicted Mean Steady State Parameters
Cmax,ss Cmin,ss AUC,ss
Dosing Regimen
(pg/mL) (pg/mL) (pg*day/mL)
200 mg LD(D14)+ QM] 89 18 1110
600 mg LD(D14)+ QM] 268 53 3329
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
83
1200 mg LD(D14)+ QM] 536 106 6659
LD = Loading dose; QM = Every 28 days.
3. Conclusions
[0286] Sifalimumab PK was best described using a 2-compartment linear model
with first order elimination. Following IV dosing, the typical clearance (CL)
and
central volume of distribution (Vc) were estimated to be 176 mL/day and 2.9 L,
respectively. The estimates of between-subject variability for CL and Vc were
28% and 31%, respectively. Patient baseline body weight, IFN gene signature
(21 genes), steroid use and sifalimumab dose were identified as significant
covariates for CL, whereas only baseline body weight was significant covariate
for Vc and Vp. Surprisingly, the above mentioned covariates were statistically
significant, but they did not explain variability in sifalimumab PK parameters
to
any relevant extent. Thus leads to the conclusion that no weight-based dosing
adjustments are necessary to administer sifalimumab. VPC results demonstrated
good predictability of the final population PK model.
[0287] Simulation results demonstrated that both fixed and body weight
based
dosing regimens yield similar median steady state concentrations (Cõ) and
variability. Accordingly, fixed sifalimumab doses of 200, 600 and 1200 mg
monthly (with a loading dose at day 14) are selected for phase II clinical
trial.
[0288] A population PK model of sifalimumab was developed and validated.
The
population PK analysis also demonstrated the feasibility of evaluating fixed
doses
of sifalimumab in phase II clinical trials.
Example 3
Pharmacokinetics and Immunogenicity of Single and Multiple Fixed Dosing
Regimens of Sifalimumab Administered Subcutaneously to SLE Patients
[0289] The pharmacokinetics (PK) and immunogenicity (IM) of single and
multiple
subcutaneous (SC) fixed doses of sifalimumab administered to adult patients
with
moderate to severe SLE was studied in a phase II clinical trial.
1. Methods
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
84
1.1 Study design
[0290] This was a multicenter, randomized, double-blind, placebo-
controlled,
Phase I la parallel study with 5 evaluable arms (1:1:2:2:2 ratio).
[0291] Criteria for inclusion of subjects receiving the antibody or antigen
binding
fragment thereof subcutaneously are summarized in clinical trial identifier
NCT00657189, accessible via the U.S. National Institutes of Health
clinicaltrials.gov database. Thus, inclusion criteria comprise: Male or female
subjects were over 18 years and below 95 years of age at the time of the first
dose of study drug; subjects met at least 4 of the 11 revised ACR
classification
criteria for SLE; subjects had positive antinuclear antibody test (ANA) at
1:80
serum dilution documented in the past or at screening; subjects had at least 1
system with a score of A or 2 systems with a score of B on the BILAG index at
screening, or have a SELENA-SLEDAI score 6; and treatment for SLE with
antimalarials, oral prednisone or another systemic corticosteroid,
mycophenolate
mofetil, methotrexate, leflunomide, azathioprine, or dapsone.
[0292] Exclusion Criteria comprised: having received MEDI-545 within 120
days
prior to screening; history of allergy or reaction to any component of the
study
drug formulation; having received the following medications within 28 days
before
randomization: systemic cyclophosphamide at any dose, cyclosporine at any
dose, thalidomide at any dose, hydroxychloroquine > 600 mg/day,
mycophenolate mofetil > 3 g/day, methotrexate > 25 mg/week, azathioprine > 3
mg/kg/day; having received fluctuating doses of the following within 28 days
before randomization: antimalarials, mycophenolate mofetil, methotrexate,
leflunomide, azathioprine, dapsone; having received leflunomide > 20mg/day in
the 6 months prior to Study Day 0; having received prednisone > 20 mg/day or
in
fluctuating doses within 14 days before randomization; having received
fluctuating doses of non-steroidal anti-inflammatory drugs within 14 days
before
randomization; treatment with any investigational drug therapy within 28 days
before randomization into the study, B cell-depleting therapies within 12
months
before randomization, or biologic therapies within 30 days or 5 half-lives of
the
biologic agent, whichever is longer, before randomization into the study; in
the
investigator's opinion, evidence of clinically significant active infection,
including
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
ongoing, chronic infection, within 28 days before randomization; a history of
severe viral infection as judged by the investigators, including severe
infections of
either cytomegalovirus or the herpes family such as disseminated herpes,
herpes
encephalitis, ophthalmic herpes; herpes zoster infection within 3 months
before
randomization; evidence of infection with hepatitis B or C virus, or human
immunodeficiency virus (HIV)-1 or HIV-2, or active infection with hepatitis A,
as
determined by results of testing at screening; vaccination with live
attenuated
viruses within 28 days before randomization; pregnancy (women, unless
surgically sterile or at least 2 years post-menopausal, had a negative serum
pregnancy test within 28 days before receiving the study drug and a negative
urine pregnancy test on days of study drug administration before receiving the
study drug); breastfeeding or lactating women; history of primary
immunodeficiency; history of alcohol or drug abuse < 1 year prior to
randomization; history of cancer (except basal cell carcinoma or in situ
carcinoma
of the cervix treated with apparent success with curative therapy > 1 year
prior to
randomization); history of active tuberculosis (TB) infection or newly
positive TB
skin test (defined as a reaction 10 mm
in diameter if not on systemic
immunosuppressive medication or 5 mm if on systemic immunosuppressive
medication history of latent TB infection without completion of an appropriate
course of treatment; elective surgery planned from the time of screening
through
Study Day 168; at screening blood tests (within 28 days before randomization),
any of the following (i) AST > 2.5 x upper limit of the normal range (ULN),
unless
caused by SLE, 9 (ii) ALT > 2.5 x ULN, unless caused by SLE, (iii) Creatinine
>
4.0 mg/dL, (iv) Neutrophils < 1,500/mm3, (v) Platelet count < 50,000/mm3;
history of any disease, evidence of any current disease (other than SLE), any
finding upon physical examination, or any laboratory abnormality that, in the
opinion of the investigator or medical monitor, may compromise the safety of
the
subject in the study or confound the analysis of the study.
[0293] The inclusion and exclusion criteria listed above are not
intended to limit
the scope of the present disclosure. A person skilled in the art would
understand
that other inclusion and/or exclusion parameters may be used to generate a
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
86
subject population such that the choice of subject does not compromise the
safety of the subject or confounds the analysis of the study.
[0294] Patients were administered up to 13 doses of either sifalimumab or
placebo. Placebo was administered to 22 patients. Eleven patients received a
single SC 100 mg fixed dose of sifalimumab. Twenty-one patients received a
monthly SC 100 mg fixed dose of sifalimumab, with the last dose being
administered on Day 84. Twenty-three patients received biweekly SC 100 mg
fixed doses of sifalimumab, with the last dose being administered on Day 84.
Ten
patients received weekly SC 100 mg fixed doses of sifalimumab, with the last
dose being administered on Day 84. Serum samples were collected for PK
concentrations and IM titers at multiple time points.
[0295] Samples were analyzed for PK and IM using respectively the validated
ELISA and ECL assays described in Example 1.
1.2 Data Analysis
[0296] Pharmacokinetic parameters were obtained using non-compartmental
methods using WinNonlin (version 5.2.1 Pharsight, Cary, NC).
2. Results
2.1 Single Dose Administration
[0297] Results corresponding to the administration of a single 100 mg fixed
dose
of sifalimumab are summarized in TABLE 7 and FIG. 9 (.). Peak concentrations
of sifalimumab occurred approximately 1 week after SC administration. Mean
apparent extravascular clearance (CL/F) was approximately 275 mL/day.
Elimination half-life was approximately 25 days. Plotted values appear in
TABLE
10.
2.2 Multiple Dose Administration
[0298] Results corresponding to the administration of multiple 100 mg fixed
doses
of sifalimumab are summarized in TABLE 8, and FIG. 9 (weekly., bi-weekly A,
or monthly V). Cmax ss and Ctrough ss increased with dosing frequency.
Accumulation of AUC and trough concentrations was 11-and 6-fold, respectively,
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
87
for weekly administration. Steady state AUC for multiple dose groups was
similar
to AUC after a single dose. Mean apparent extravascular steady state clearance
(CLõ/F) ranged from 206 to 282 mL/day across the dosing groups. Mean half-life
ranged from 28 to 33 days. Apparent volume of distribution during terminal
phase
after non-intravenous administration (Vz/F) ranged from 8 L to 11 L.
2.3 Immunogenicity
[0299] Eight out of 65 patients (12.3%) receiving investigational product
tested
positive for presence of anti-sifalimumab antibodies (FIG. 10). There was no
increase in incidence of immunogenicity across dosing regimens. Two out of 22
patients receiving placebo (9.1%) were M+, i.e., they tested positive for the
presence of anti-sifalimumab antibodies (titer range 10 to 40). None of the
eleven
patients receiving a single dose of sifalimumab were M. In contrast, three out
of
ten patients following a monthly dosing regime (30%) were M+ (titer range 20
to
160), three out of 23 patients following a bi-weekly dosing regime (13%) were
M+
(titer range 20 to 160) and two out of 21 patients (9.5%) following a weekly
dosing regime (9.5%) were M+ (titer range 20 to 320).
[0300] Presence of anti-sifalimumab antibody did not impact
serum concentration-time profiles of sifalimumab. Sifalimumab had a
safety/tolerability profile similar to that of placebo at the SC doses tested.
2.4 Effect on IFN Gene Signature
[0301] Pharmacodynamics (PD) markers can be used in methods of treating
patients with a therapeutic agent that binds to and modulates IFN-alpha
activity
such as sifalimumab. See, e.g., U.S. Patent Appl. No. 2010-0143372, which is
hereby incorporated herein by reference in its entirety. Specifically, Example
7 in
the 2010-0143372 publication describes 21 genes that can be used as PD
markers: IF144, IF127, IF144L, NAPTP, LAMP3, LY6E, RSAD2, HERC5, IF16,
ISG15, 0A53, RTP4, IFIT1, MX1, SIGLEC1, 0A52, USP18, OAS1, EPSTI1,
PLSCR1 and IFRG28. Additionally, the 2010-0143372 publication provides
methods for how to measure the levels of these 21 gene markers, e.g.,
Affymetrix
arrays and Fluidigm dynamic arrays.
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
88
[0302] Only the once-weekly dose of sifalimumab generated a sufficient PK
exposure to induce a sustained IFN gene signature decrease over time (FIG.
11).
In particular, weekly dosing produced suppression of the gene signature up to
40%.
3. Conclusion
[0303] Comparison of pharmacokinetic exposures of sifalimumab to previous
intravenous studies indicated SC bioavailability of approximately 75%. PK of
sifalimumab following SC administration was linear and dose-proportional.
Pharmacokinetic parameters of sifalimumab following SC dosing were typical of
a
monoclonal antibody without a significant target sink. The overall incidence
of
anti-sifalimumab antibodies was 12%, however, the presence of anti-sifalimumab
antibody did not impact the pharmacokinetics of sifalimumab.
TABLE 7
Sifalimumab Single Dose PK Parameters
Parameter Single 100 mg SC Dose
(n = 10)
T (day) 5.78 (62.2)
max
C (pg/mL) 12.68 (68.4)
max
AUC (pg=day/mL) 420.6 (58.3)
last
AUC (pg=day/mL) 477.6 (57.2)
AUC Extrapolation (`)/0) 12.5 (33.2)
CL/F (mL/day) 275 (57.0)
Half-life (day) 24.6 (37.8)
V /F (L) 8.32 (38.4)
Data from PK evaluable patients presented as mean (%CV).
TABLE 8
Sifalimumab Multiple Dose PK Parameters
Parameter 100 mg SC Weekly 100 mg SC Bi-weekly 100 mg SC Monthly
(n = 8) (n = 6) (n = 6)
CA 02836926 2013-11-20
WO 2012/162367
PCT/US2012/039098
89
T (day) 4.29 (51.6) 4.30 (59.0) 5.64 (35.9)
max ss
C (pg/mL) 65.05 (42.7) 39.10 (24.2) 23.96 (39.8)
max ss
AUC (pg=day/mL) 442.7 (44.0) 495.2 (14.4) 483.1 (32.6)
T SS
(pg/mL) 58.80 (36.2) 30.10 (31.7) 10.64 (39.6)
trough ss
CL /F (mL/day) 282 (40.4) 206 (16.5) 227 (33.1)
ss
Half-life (day) 28.2 (22.8) 28.2 (32.7) 33.3 (41.6)
V /F (L) 11.1 (36.9) 8.19(26.7) 11.0(51.9)
Data from PK evaluable patients presented as mean (CV%).
Parameters calculated after dosing on Day 84
- 90 -
TABLE 9
o
t..,
=
,-,
t..,
,-,
cA
Variable = PKConc
w
CP-152
cA
--.1
STUDYID
CP152
NomTime
TRTGRP 0.00 0.04 0.08 9.00 13.00
14.00 28.00 42.00 47.00 56
MEDI-545 0.3 MG/KG
N 32 26 26 Missing
Missing 25 26 24 Missing 24
Mean
0.000 10.092 9.263 Missing Missing 2.367 4.315 4.750
Missing 5.995 n
SD
0.000 4.493 4.263 Missing Missing 1.208 2.264 2.442
Missing 3.223 o
1.)
Min 0.00 0.00 0.00
Missing Missing 0.00 0.00 1.44 Missing 1.69 co
us)
Median
0.000 10.185 8.405 Missing Missing 2.470 3.920 4.345
Missing 5.785 o,
ko
Max 0.00 22.67
22.01 Missing Missing 4.91 9.04 11.01 Missing 11.76 1.)
o,
CV% Missing 44.5
46.0 Missing Missing 51.0 52.5 51.4 Missing 53.7 1.)
o
Geometric Mean
Missing Missing Missing Missing Missing Missing Missing 4.210 Missing
5.134 usil-
MEDI-545 1.0 MG/KG1
H
H
N 26 25 25 Missing
Missing 24 24 23 Missing 23 1
1.)
Mean
0.867 29.361 28.199 Missing Missing 7.957 14.178 15.378
Missing 18.247 o
SD
3.991 13.410 8.008 Missing Missing 4.103 10.808 6.272
Missing 7.277
Min 0.00 0.00 16.85
Missing Missing 3.38 6.02 6.13 Missing 7.92
Median
0.000 28.320 26.070 Missing Missing 6.765 11.195 13.970
Missing 18.290
Max
20.32 66.43 51.65 Missing Missing 19.12 59.58 27.48
Missing 35.22
CV% 460.6 45.7 28.4
Missing Missing 51.6 76.2 40.8 Missing 39.9
Geometric Mean Missing Missing 27.206
Missing Missing 7.137 12.106 14.116 Missing 16.884 IV
n
MEDI-545 10.0 MG/KG
1-3
N 44 43 43 1 Missing
40 38 39 Missing 36
ri)
Mean
0.000 210.824 217.006 66.260 Missing 53.094 80.863
90.803 Missing 110.835 n.)
o
1-,
SD
0.000 85.935 84.895 Missing Missing 25.827 40.513
43.404 Missing 47.480 n.)
-1
Min 0.00 14.47
14.53 66.26 Missing 8.12 17.45 27.43 Missing 3.51 c.")
o
Median
0.000 211.670 227.300 66.260 Missing 51.595 72.860
83.950 Missing 102.605
o
Max
0.00 379.16 382.56 66.26 Missing 114.52 174.90 178.60
Missing 210.80 oe
- 91 -
CV% Missing 40.8 39.1
Missing Missing 48.6 50.1 47.8 Missing 42.8
Geometric Mean Missing 185.949 191.651
66.260 Missing 46.223 70.749 80.733 Missing 95.020 0
n.)
MEDI-545 3.0 MG/KG
o
1-,
n.)
N 25 25 25
Missing Missing 24 25 26 Missing 27
o
Mean
0.000 99.292 96.112 Missing Missing 22.367 28.007
40.381 Missing 43.022 i..)
SD
0.000 41.106 37.254 Missing Missing 10.808 15.212
22.740 Missing 23.770 o
--.1
Min 0.00
35.90 28.39 Missing Missing 6.79 5.66 12.43 Missing 12.31
Median
0.000 94.830 95.100 Missing Missing 20.050 28.190
32.905 Missing 38.180
Max
0.00 184.95 178.54 Missing Missing 47.56 70.64 98.31
Missing 116.83
CV% Missing 41.4 38.8
Missing Missing 48.3 54.3 56.3 Missing 55.3
Geometric Mean Missing 90.440 88.707
Missing Missing 19.868 24.042 34.986 Missing 37.837
I TRTGRP 70.00 84.00 96.00 98.00 112.00
113.00 121.00 126.00 140.00 141.00 154.00 n
MEDI-545 0.3 MG/KG
o
24 23 Missing 23 23 Missing 1 23
23 Missing 23 co"
6.080 6.710 Missing 6.498 6.733 Missing 0.000 6.930 6.231 Missing 6.256 us)
o,
ko
3.522 4.656 Missing 4.477 3.740 Missing Missing 4.617 3.291 Missing 3.201
1.)
o,
0.00 0.00 Missing 0.00 0.00 Missing 0.00 0.00
0.00 Missing 0.00 1.)
5.760 5.580 Missing 5.750 6.220 Missing 0.000 6.490 5.940 Missing 6.670 0
H
12.34 18.66 Missing 18.52 15.04 Missing 0.00 20.12 13.76 Missing
13.66 us)
1
H
57.9 69.4 Missing 68.9 55.6 Missing Missing 66.6
52.8 Missing 51.2 H
I
Missing Missing Missing Missing Missing Missing Missing Missing Missing
Missing Missing o"
MEDI-545 1.0 MG/KG
21 23 Missing 22 23 Missing Missing 20 22
Missing 20
18.521 18.665 Missing 19.625 20.676 Missing Missing 22.715 21.077 Missing
20.286
6.361 7.704 Missing 8.486 10.770 Missing Missing 13.577 10.473 Missing 8.708
9.47 3.12 Missing 7.04 5.06 Missing Missing 4.97
4.27 Missing 4.66
19.160 17.720 Missing 18.280 19.550 Missing Missing 19.255 17.815 Missing
19.255 IV
31.94 36.57 Missing 38.25 58.53 Missing Missing 57.11 50.45 Missing 38.43 n
,-i
34.3 41.3 Missing 43.2 52.1 Missing Missing 59.8
49.7 Missing 42.9
17.485 16.812 Missing 17.935 18.372 Missing Missing 19.430 18.714 Missing
18.346 ri)
n.)
o
MEDI-545 10.0 MG/KG
n.)
36 38 Missing 36 37 1 Missing 33 34
Missing 34 -1
133.145 138.676 Missing 138.143 138.298 196.480 Missing 138.248 130.337
Missing 123.058 o
o
o
66.232 63.355 Missing 82.577 72.621 Missing Missing 72.618 83.074 Missing
86.410 oe
0.00 26.09 Missing 0.00 21.41 196.48 Missing 8.24 5.40 Missing
17.07
- 92 -
117.345 131.150 Missing 141.825 128.690 196.480 Missing 138.630 118.180
Missing 92.300
285.80 270.67 Missing 359.67 446.72 196.48 Missing 293.67 370.79 Missing
379.66 0
n.)
49.7 45.7 Missing 59.8 52.5 Missing Missing 52.5
63.7 Missing 70.2
1-,
Missing 123.391 Missing Missing 122.299 196.480 Missing 112.943 96.644 Missing
95.710 n.)
1-,
MEDI-545 3.0 MG/KG
o
n.)
27 24 Missing 24 27 Missing Missing 23
20 1 21 o
--.1
45.782 35.673 Missing 43.089 38.404 Missing Missing 47.260 47.215 97.700
43.872
42.551 22.427 Missing 26.721 20.672 Missing Missing 27.640 19.570 Missing
24.521
14.02 0.00 Missing 5.21 9.31
Missing Missing 10.88 11.29 97.70 5.22
33.990 34.360 Missing 35.770 34.690 Missing Missing 39.060 46.500 97.700
40.240
240.14 114.21 Missing 130.61 82.73 Missing Missing 125.58 80.46 97.70 100.60
92.9 62.9 Missing 62.0 53.8 Missing Missing 58.5
41.4 Missing 55.9
37.292 Missing Missing 35.931 32.839 Missing Missing 40.795 42.532 97.700
36.410 n
I TRTGRP 155.00 160.00 161.00 168.00 174.00
182.00 182.04 182.08 185.00 189.00 195.00 o
1.)
co
MEDI-545 0.3 MG/KG
us)
61
lo
Missing Missing Missing 23 1 22 21 21 19
20 Missing 1.)
o,
Missing Missing Missing 6.376 0.000 6.761 16.730 15.117 11.993 9.961 Missing
1.)
Missing Missing Missing 2.757 Missing 2.996 7.995 5.830 4.862 3.371 Missing
0
H
Missing Missing Missing 0.00 0.00 0.00 0.00 0.00
5.62 4.90 Missing us)
I
H
Missing Missing Missing 6.070 0.000 7.425 14.360 14.990 11.540 9.180 Missing
H
I
Missing Missing Missing 11.48 0.00 12.35 34.52 27.60 21.90
18.41 Missing o"
Missing Missing Missing 43.2 Missing 44.3 47.8 38.6 40.5
33.8 Missing
Missing Missing Missing Missing Missing Missing Missing Missing 11.163 9.466
Missing
MEDI-545 1.0 MG/KG
Missing Missing Missing 22 Missing 21 21 21 20 22
Missing
Missing Missing Missing 19.994 Missing 18.793 46.214 44.425 32.006 23.439
Missing
Missing Missing Missing 11.703 Missing 9.483 18.614 17.557 17.999 13.965
Missing IV
Missing Missing Missing 4.28 Missing 4.58 23.82 17.95
0.00 0.00 Missing n
,-i
Missing Missing Missing 18.780 Missing 18.210 36.220 39.590 29.290 20.905
Missing
Missing Missing Missing 63.04 Missing 52.00 91.34 83.13 87.45 73.32 Missing
ci)
n.)
o
Missing Missing Missing 58.5 Missing 50.5 40.3 39.5 56.2
59.6 Missing
n.)
Missing Missing Missing 17.440 Missing 16.891 43.063 41.342 Missing Missing
Missing -1
MEDI-545 10.0 MG/KG
o
o
o
Missing Missing Missing 35 Missing 34 25 26 32
33 Missing oe
Missing Missing Missing 124.736 Missing 127.904 377.440 406.550 230.993
202.535 Missing
- 93 -
Missing Missing Missing 85.858 Missing 87.376 123.078 160.702 113.038 105.993
Missing
Missing Missing Missing 17.23 Missing 16.04 212.18 223.72 13.58 5.71
Missing 0
n.)
Missing Missing Missing 114.870 Missing 116.635 362.740 379.785 246.100
206.640 Missing
1-,
Missing Missing Missing 377.85 Missing 318.62 615.91 970.99 465.88 475.09
Missing n.)
1-,
Missing Missing Missing 68.8 Missing 68.3 32.6 39.5 48.9
52.3 Missing cA
n.)
Missing Missing Missing 96.182 Missing 95.198 358.791 381.682 181.495 155.792
Missing cA
--.1
MEDI-545 3.0 MG/KG
Missing 1 1 22 Missing 20
19 19 16 22 1
Missing 2.860 5.580 47.281 Missing 53.743 138.023 140.262 113.571 70.038
20.480
Missing Missing Missing 26.448 Missing 29.582 62.191 74.014 81.401 47.081
Missing
Missing 2.86 5.58 6.78 Missing
7.20 51.26 48.40 28.42 19.13 20.48
Missing 2.860 5.580 45.375 Missing 54.155 130.310 114.120 84.925 61.880 20.480
Missing 2.86
5.58 101.36 Missing 122.84 278.99 332.73 317.73 213.48
20.48 n
Missing Missing Missing 55.9 Missing 55.0 45.1 52.8 71.7
67.2 Missing
Missing 2.860 5.580 38.588 Missing 44.482 124.958 125.412 92.032 57.005 20.480
o
1.)
I TRTGRP
co
us)
o,
MEDI-545 0.3 MG/KG 196.00 198.00 201.00 208.00 210.00
238.00 248.00 250.00 266.00 286.00 294.00 ko
1.)
o,
1.)
21 Missing Missing 1 20 21 Missing Missing
20 Missing 19 o
H
us)
7.353 Missing Missing 3.830 5.156 2.173 Missing Missing 1.283 Missing 0.208
I
H
3.293 Missing Missing Missing 2.264 1.822 Missing Missing 1.411 Missing 0.665
H
1
0.00 Missing Missing 3.83 2.09 0.00 Missing Missing
0.00 Missing 0.00 1.)
o
7.160 Missing Missing 3.830 4.960 1.980 Missing Missing 0.695 Missing 0.000
15.64 Missing Missing 3.83 11.40 5.73 Missing Missing 3.61
Missing 2.67
44.8 Missing Missing Missing 43.9 83.8 Missing Missing
110.0 Missing 319.2
MEDI-545 1.0 MG/KG Missing Missing Missing
3.830 4.708 Missing Missing Missing Missing Missing
Missing
22 Missing Missing Missing 22 23 Missing Missing 22 1 21
IV
n
17.751 Missing Missing Missing 9.783 4.345 Missing Missing 1.725 0.000 1.021
1-3
10.906 Missing Missing Missing 5.617 3.610 Missing Missing 1.998 Missing 1.253
ci)
0.00 Missing Missing Missing 0.00 0.00 Missing Missing 0.00 0.00
0.00 n.)
o
17.665 Missing Missing Missing 10.470 3.520 Missing Missing 1.635 0.000 0.000
n.)
46.89 Missing Missing Missing 21.93 14.35 Missing Missing 7.48 0.00
3.83 -1
61.4 Missing Missing Missing 57.4 83.1 Missing
Missing 115.8 Missing 122.7 o
MEDI-545 10.0 MG/KG
Missing Missing Missing Missing Missing Missing
Missing Missing Missing Missing Missing oe
- 94 -
0
34 Missing Missing Missing 34 33 Missing 1 32 Missing
33 n.)
o
155.799 Missing Missing Missing 76.119 29.602 Missing 0.000 14.674 Missing
6.088
n.)
102.784 Missing Missing Missing 56.379 30.841 Missing Missing 14.029 Missing
7.600
o
3.06 Missing Missing Missing 1.83 0.00 Missing 0.00 0.00
Missing 0.00 n.)
152.500 Missing Missing Missing 66.290 22.090 Missing 0.000 12.210 Missing
4.950 o
--.1
388.75 Missing Missing Missing 186.81 147.17 Missing 0.00 62.35 Missing
35.80
66.0 Missing Missing Missing 74.1 104.2 Missing Missing
95.6 Missing 124.8
I TRTGRP
103.647 Missing Missing Missing 47.904 Missing Missing
Missing Missing Missing Missing
MEDI-545 3.0 MG/KG 196.00 198.00 201.00 208.00 210.00
238.00 248.00 250.00 266.00 286.00 294.00
20 Missing 1 Missing 18 21 1 Missing 20
Missing 18
48.320 Missing 4.400 Missing 27.488 11.900 8.160 Missing 5.619 Missing 2.422
n
24.729 Missing Missing Missing 13.932 9.746 Missing Missing 6.392 Missing
2.917 o
1.)
10.74 Missing 4.40 Missing 5.70 0.00 8.16 Missing
0.00 Missing 0.00 co
us)
48.405 Missing 4.400 Missing 28.060 9.800 8.160 Missing 4.300 Missing 2.025
o,
ko
86.53 Missing 4.40 Missing 59.11 38.62 8.16 Missing
22.83 Missing 12.24
51.2 Missing Missing Missing 50.7 81.9 Missing
Missing 113.8 Missing 120.5 1.)
o
41.052 Missing 4.400 Missing 23.762 Missing 8.160 Missing Missing Missing
Missing H
us)
I
H
H
I
IV
0
TABLE 10
STUDYID
CP179 IV
n
NomTime
1-3
TRTGRP 0 3 7 14 21 28 35 36
41 42 49 56
ci)
n.)
MEDI-545 100MG ONCE
=
1-,
N 11 10 9 10 10 10 9
Missing Missing 9 8 8 n.)
-I
Mean 0.000 11.552 9.242 9.115 7.336
5.723 5.257 Missing Missing 4.779 4.063 3.409 c,.)
o
o
SD 0.000 9.161 2.224 4.420 4.496 2.871
2.478 Missing Missing 2.137 2.063 2.039 o
oe
Min 0.00 3.31 5.14 4.36 3.15 1.37
3.14 Missing Missing 2.54 1.51 1.27
- 95 -
Median 0.000 9.795 9.620 8.690 6.430 4.885
3.670 Missing Missing 4.390 4.055 3.095
Max 0.00 36.69 12.37 19.58 18.16 10.96
9.35 Missing Missing 8.03 7.43 6.52 0
n.)
CV% Missing 79.3 24.1 48.5 61.3 50.2
47.1 Missing Missing 44.7 50.8 59.8
1-,
Geometric Mean Missing 9.629 8.974 8.295 6.381
5.016 4.797 Missing Missing 4.356 3.588 2.846 n.)
1-,
MEDI-545 100MG Q1WK
o
n.)
N 21 19 21 18 19 20 20
Missing 1 16 18 18 o
--.1
Mean 0.000 8.543 9.785 16.541 21.436
27.827 31.182 Missing 13.190 36.984 37.237 36.583
SD 0.000 5.164 4.193 6.037 7.204 10.194
13.329 Missing Missing 14.132 16.891 17.258
Min 0.00 2.14 4.40 8.66 8.43 13.58
7.25 Missing 13.19 18.50 7.69 5.86
Median 0.000 7.480 9.570 17.570 20.890
26.440 29.665 Missing 13.190 34.045 34.080 35.655
Max 0.00 21.87 22.28 27.21 34.33 50.09
54.36 Missing 13.19 59.08 64.61 74.60
CV% Missing 60.4 42.9 36.5 33.6 36.6
42.7 Missing Missing 38.2 45.4 47.2
Geometric Mean Missing 7.152 9.030 15.448 20.268
26.119 28.079 Missing 13.190 34.554 32.588 31.355 n
MEDI-545 100MG
Q2WKS
o
1.)
N 22 21 22 21 18 17 15 1
Missing 17 17 16 co
us)
o,
Mean 0.119 9.856 10.660 8.888 16.981
14.712 18.436 5.470 Missing 15.331 20.364 16.719 ko
1.)
SD 0.559 5.352 5.160 4.112 7.866 8.329
6.904 Missing Missing 8.322 9.643 8.055 o,
Min 0.00 2.68 3.74 2.94 6.77 5.83
5.42 5.47 Missing 2.51 4.05 4.18 1.)
0
H
Median 0.000 8.680 9.030 7.700 16.145
11.770 20.030 5.470 Missing 15.790 20.330 16.450 La
1
Max 2.62 21.34 24.42 18.65 35.47 37.58
27.64 5.47 Missing 34.89 38.81 32.87
H
I
CV% 469.0 54.3 48.4 46.3 46.3 56.6
37.4 Missing Missing 54.3 47.4 48.2 1.)
Geometric Mean Missing 8.518 9.582 8.075 15.375
13.034 16.803 5.470 Missing 12.798 17.485 14.540 o
MEDI-545 100MG
Q4WKS
N 10 9 9 10 9 10 9
Missing Missing 10 10 10
Mean 0.000 8.781 10.136 9.953 8.440 7.251
14.196 Missing Missing 13.297 10.159 8.411
SD 0.000 4.267 3.794 3.283 2.166 1.531
6.100 Missing Missing 5.599 4.694 3.460
Min 0.00 3.88 5.39 6.38 5.09 4.46
4.66 Missing Missing 3.57 1.96 2.65 IV
n
Median 0.000 8.630 10.260 10.010 8.230
7.325 14.560 Missing Missing 13.600 10.080 8.715 1-3
Max 0.00 16.58 17.59 17.07 12.44 10.30
22.86 Missing Missing 23.17 19.07 14.47
ri)
CV% Missing 48.6 37.4 33.0 25.7 21.1
43.0 Missing Missing 42.1 46.2 41.1 n.)
o
1-,
Geometric Mean Missing 7.849 9.503 9.510 8.195
7.099 12.867 Missing Missing 11.962 8.869 7.622 n.)
-1
I TRTGRP 63 70 77 84 87 91 98 112
126 140 168 I o
=
o
oe
MEDI-545 100MG ONCE
- 96 -
N 6 5 5 5 6 5 4 1
0 0 0
Mean 3.377 3.940 3.174 2.316 1.775 1.886
1.518 1.330 Missing Missing Missing 0
n.)
SD 1.801 0.935 0.712 1.003 0.500 0.511
0.201 Missing Missing Missing Missing o
1-,
Min 1.36 2.68 2.37 1.52 1.27 1.46
1.35 1.33 Missing Missing Missing n.)
1-,
Median 3.495 3.810 3.140 2.060 1.685 1.820
1.460 1.330 Missing Missing Missing o
n.)
Max 5.30 4.87 4.10 4.02 2.73 2.71
1.80 1.33 Missing Missing Missing o
--.1
CV% 53.3 23.7 22.4 43.3 28.2 27.1
13.3 Missing Missing Missing Missing
Geometric Mean 2.929 3.846 3.110 2.174 1.725 1.836
1.508 1.330 Missing Missing Missing
MEDI-545 100MG Q1WK
N 17 17 17 15 13 14 16 16
16 16 14
Mean 41.655 41.936 43.665 49.877 51.812
49.356 38.062 22.339 16.376 12.427 7.001
SD 16.144 27.702 19.666 27.647 26.100
21.776 19.036 10.016 9.374 7.767 4.619
Min 18.56 2.15 1.93 20.83 19.34 17.50
14.05 6.15 3.38 1.55 1.45 n
Median 35.850 35.930 41.450 39.340 45.230
45.350 33.455 22.275 15.700 11.265 5.975
o
Max 71.11 93.34 87.62 115.29 116.84
94.23 80.06 38.08 39.20 27.83 18.26 1.)
co
CV% 38.8 66.1 45.0 55.4 50.4 44.1
50.0 44.8 57.2 62.5 66.0 us)
o,
Geometric Mean 38.947 28.364 36.331 44.037 46.256
44.735 33.746 19.825 13.649 9.729 5.669 ko
1.)
MEDI-545 100MG
o,
Q2WKS
1.)
0
H
N 17 16 15 16 13 17 17 16
14 12 10 us)
I
Mean 19.206 19.028 23.946 19.311 28.338
23.594 24.095 14.793 11.443 7.432 5.179 H
H
I
SD 7.915 8.785 12.946 10.670 16.436
13.104 15.020 9.576 8.233 6.060 4.151 1.)
Min 6.72 6.20 5.51 5.77 5.32 5.56
3.34 3.01 3.20 1.66 1.96 o
Median 20.820 20.650 28.860 16.235 24.410
21.870 19.260 11.280 8.650 4.855 4.115
Max 29.70 32.25 42.76 38.22 55.21 53.31
60.39 37.78 26.89 20.67 15.93
CV% 41.2 46.2 54.1 55.3 58.0 55.5
62.3 64.7 71.9 81.5 80.2
Geometric Mean 17.337 16.613 19.980 16.383 23.321
20.021 19.436 12.038 8.945 5.395 4.195
MEDI-545 100MG
Q4WKS
IV
n
N 10 10 10 10 10 10 8 10
8 8 6 1-3
Mean 16.795 13.237 10.233 12.729 16.217
17.747 15.818 8.434 7.629 4.354 3.522
ri)
SD 10.001 6.304 5.270 11.012 7.734
10.748 9.465 4.445 3.316 2.183 2.065 n.)
o
1-,
Min 3.12 3.48 2.25 1.68 6.39 4.75
3.86 2.56 3.90 1.53 1.65 n.)
-1
Median 16.375 14.475 10.675 11.615 13.340
13.570 13.245 7.790 6.935 4.525 3.015 c,.)
o
Max 35.67 23.28 18.24 39.74 31.35 34.85
31.57 18.86 14.56 7.79 6.58 =
o
CV% 59.5 47.6 51.5 86.5 47.7 60.6
59.8 52.7 43.5 50.1 58.7 oe
- 97 -
Geometric Mean 13.657 11.516 8.670 9.022 14.655 14.868
13.263 7.444 7.087 3.802 3.031
0
o
1.)
us)
61"
0
0
,4z
,4z
oe
CA 02836926 2013-11-20
WO 2012/162367 PCT/US2012/039098
- 98 -
TABLE 11
Expected PK Exposure in 40 kg Patient Population
Cmax,ss Cmin,ss
5th 95th 5th 95th
Dosing Regimen Percentile Percentile Percentile Percentile
0.3 mg/kg (Q14D) 6.68 15.25 2.29 8.11
1 mg/kg (Q14D) 21.25 49.29 6.8 24.65
3 mg/kg (Q14D) 61.84 143.96 18.47 68.11
mg/kg (Q14D) 200.29 466.6 55.03 210.1
30 mg/kg (Q14D) 584.94 1359.5 148.37 582.22
200 mg [LD(D14)+QM] 80.26 189.51 9.435 48.035
600 mg [LD(D14)+QM] 240.78 568.55 28.305 144
1200 mg [LD(D14)+QM] 481.57 1137.1 56.61 288.21
Expected PK Exposure in 120 kg Patient Population
Cmax,ss Cmin,ss
5th 95th 5th 95th
Dosing Regimen Percentile Percentile Percentile Percentile
0.3 mg/kg (Q14D) 11.64 26.62 3.9 13.95
1 mg/kg (Q14D) 37.02 85.14 11.52 42.55
3 mg/kg (Q14D) 107.61 248.69 31.19 117.59
10 mg/kg (Q14D) 348.51 806.2 92.58 357.74
30 mg/kg (Q14D) 1016.4 2361 249.13 993.96
200 mg [LD(D14)+QM] 46.43 110.61 5.415 28.23
600 mg [LD(D14)+QM] 139.3 331.84 16.245 84.71
1200 mg [LD(D14)+QM] 278.6 663.68 32.49 169.42
