Note: Descriptions are shown in the official language in which they were submitted.
PATENT
Attorney Docket No.: 14002.6004-00000
METHODS AND USES OF COMPOUNDS FOR TREATMENT OF
CHORIOCAPILLARY DISORDERS
FIELD
[0001] The present invention is directed to improved therapies for
diseases that
disrupt the choriocapillaris vasculature.
BACKGROUND
[0002] Impacting 30-50 million people worldwide, age-related macular
degeneration (AMD) is the leading cause of vision loss in the industrialized
world (1). By
2040, this number is expected to grow to over 288 million, representing an
ever-
increasing burden on the global health system (2). Despite decades of
research, the
pathophysiology of AMD remains incompletely understood. Several contributing
factors
have been identified, including inflammation, complement activation, oxidative
stress
and hypoxia (3). Clinically, early AMD is characterized by subretinal drusen
deposition
and pigment alterations in the retinal pigmented epithelium (RPE)(4). Defects
in
choroidal blood flow are observed, and reduced perfusion of the
choriocapillaris, a
unique vascular bed which supplies the outer retina (FIG. 1), is correlated
with disease
progression (5-7), suggesting a potential link between retinal ischemia and
AMD
[0003] Late-stage AMD is divided into two types: Neovascular (wet) and
atrophic (dry), which can coexist in the same eye. Dry AMD is characterized by
2
CA 3075146 2020-03-11
= PATENT
Attorney Docket No.: 14002.6004-00000
choriocapillaris atrophy combined with RPE degeneration and photoreceptor
dysfunction. This syndrome is known as geographic atrophy and no treatment is
currently available, although it is responsible for the majority of AMD-
associated vision
loss (8). Wet AMD is characterized by choroidal neovascularization (CNV)
caused by
excessive production of vascular endothelial growth factor (VEGF), leading to
intra- and
subretinal fluid leakage (9). Studies of neovascular membranes in wet AMD have
revealed activation of hypoxia inducible factor (HIF) signaling (10, 11). As
the majority of
CNV occurs in regions with poor choroidal perfusion (12-14), this suggests
that RPE
hypoxia due to reduced choriocapillaris blood flow leads to excessive VEGF
production
and ultimately CNV and CNV associated fluid leakage. VEGF is central to
disease
progression and wet AMD is treated with intravitreal VEGF inhibitors which
have
transformed disease prognosis. However, these drugs are not curative, nor are
they
effective for all patients. Following repeated injections, 10-50% of patients
develop
reduced visual acuity and choriocapillaris or RPE atrophy (15), possibly due
to the
necessity of VEGF signaling for choriocapillaris and photoreceptor maintenance
(16-
19). Supporting this hypothesis, the GAIT and IVAN trials have suggested that
increased VEGF inhibitor treatment intensity is associated with higher
incidence of
geographic atrophy after two years, underscoring the importance of VEGF for
choriocapillaris homeostasis and the need for alternative therapeutic options
(20, 21).
[0004] The potential links between VEGF inhibition and geographic
atrophy
highlight the need to recognize clinical features distinguishing nonresponsive
patients
3
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
before subjecting them to increasing doses of anti-VEGF therapy. Polypoidal
choroidal
vasculopathy (PCV), a subtype of neovascular AMD particularly prevalent in
African-
American and Asian patient populations, may be one such group (22, 23). PCV is
characterized by a disorganized network of dilated choroidal vessels and sub-
RPE
polypoid lesions which lead to subretinal hemorrhage and lipid exudate (24-
26). Small
trials have found poor results with ranibizumab (27, 28) and although improved
outcomes using aflibercept have been reported (29), recent large-scale trials
have
shown that anti-VEGF monotherapy leads to incomplete polyp closure (30-32).
Intriguingly, while the cause of PCV is unknown, a recent study identified
several SNPs
in the ANGPT2 locus in an Asian PCV cohort (33), supporting the possibility
that
Angiopoietin-TIE2 signaling is implicated in this disease.
[0005] The choriocapillaris is a polarized, fenestrated vascular bed
with a
unique lobular architecture. Unlike retinal vessels, which form through
vasculo- and
angiogenesis, the choriocapillaris develops through the hemo-vasculogenesis,
in which
common precursors give rise to both blood and endothelial cells arranged in
blood-
island-like structures (34, 35). Little is known about the pathways regulating
this
developmental process. However, VEGF signaling is essential for
choriocapillaris
development (16), and in adult mice, RPE-produced VEGF is required for
choriocapillaris and photoreceptor maintenance (19, 36). This ongoing
requirement for
VEGF highlights the relationship between RPE, choriocapillaris and retinal
health, and
emphasizes the potential impact of pharmacologic VEGF inhibition. The
importance of
4
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
VEGF for choriocapillaris homeostasis underscores the need for novel AMD
therapeutic
targets and strategies that suppress neovascularization while sparing
choriocapillaris
function. While anti-VEGF drugs have proven effective in the wet form of AMD,
they are
not curative and fail to provide relief to some patients, particularly those
with atypical
AMD or during the end stages of disease progression. Furthermore, as
pathological
VEGF production is a consequence rather than the primary pathogenic factor in
AMD,
there is an urgent, unmet need for new therapeutic approaches that improve
choroidal
perfusion, slow disease progression, and protect the choriocapillaris
vasculature during
anti-VEGF therapy.
SUMMARY
[0006] The present invention is directed, in general, to therapies for
diseases
that disrupt choriocapillaris, including AMD, and to mice carrying at least
one
angiopoietin 1 allele and at least one Wnt-Cre allele. Several of the various
features of
the invention will be described hereinafter. It is to be understood that the
invention is
not limited in its application to the details set forth in the following
embodiments, claims,
description and figures. The invention is capable of other embodiments and of
being
practiced or carried out in numerous other ways.
[0007] Embodiment 1: A method of treating age-related macular
degeneration
(AMD) or PCV in a subject in need thereof, comprising administering to the
subject a
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
therapeutically effective amount of a Tie2 activator, optionally in
combination with anti-
VEGF therapy.
[0008] Embodiment 2: A method of treating AMD or PCV in a subject in
need
thereof, comprising administering to the subject a therapeutically effective
amount of a
VE-PTP inhibitor, optionally in combination with anti-VEGF therapy.
[0009] Embodiment 3: A method of improving vision in a subject in need
thereof
comprising administering to the subject a therapeutically effective amount of
a Tie2
activator and/or a VE-PTP inhibitor, optionally in combination with anti-VEGF
therapy,
wherein the subject has AMD, preferably variants of choroid polypoid
vasculopathy,
geographic atrophy associated with anti-VEGF therapy, and any other loss of
choroid
perfusion (e.g. diabetic eye disease).
[0010] Embodiment 4: A method of increasing and/or improving
choriocapillaris
vascularization in a subject in need thereof, the method comprising
administering to the
subject a therapeutically effective amount of a Tie2 activator, optionally in
combination
with anti-VEGF therapy, wherein choriocapillaris vascularization is increased
and/or
improved in the subject.
[0011] Embodiment 5: A method of increasing and/or improving
choriocapillaris
vascularization in a subject in need thereof, the method comprising
administering to the
subject a therapeutically effective amount of a VE-PTP inhibitor optionally in
combination with anti-VEGF therapy, wherein choriocapillaris vascularization
is
increased and/or improved in the subject.
6
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0012] Embodiment 6: A method of reducing choriocapillaris dropout
during anti-
VEGF therapy in a subject in need thereof, the method comprising administering
to the
subject a therapeutically effective amount of a Tie2 activator optionally in
combination
with anti-VEGF therapy, wherein choriocapillaris dropout is reduced
[0013] Embodiment 7: A method of reducing choriocapillaris dropout
during anti-
VEGF therapy in a subject in need thereof, the method comprising administering
to the
subject a therapeutically effective amount of a VE-PTP inhibitor optionally in
combination with anti-VEGF therapy, wherein choriocapillaris dropout is
reduced.
[0014] Embodiment 8: A method of improving choroidal perfusion in a
subject in
need thereof having age-related macular degeneration (AMD), the method
comprising
administering to the subject a therapeutically effective amount of a Tie2
activator,
wherein choroidal perfusion is increased; and optionally wherein the subject
is being
treated with anti-VEGF therapy.
[0015] Embodiment 9: A method of improving choroidal perfusion in a
subject in
need thereof having age-related macular degeneration (AMD), the method
comprising
administering to the subject a therapeutically effective amount of a VE-PTP
inhibitor,
wherein choroidal perfusion is increased; and optionally wherein the subject
is being
treated with anti-VEGF therapy.
[0016] Embodiment 10: A method of protecting the choriocapillaris
vasculature
in a subject in need thereof having AMD, the method comprising administering
to the
subject a therapeutically effective amount of a Tie2 activator, wherein the
7
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
choriocapillaris vasculature is protected; and optionally wherein the subject
is being
treated with anti-VEGF therapy.
[0017] Embodiment 11: A method of protecting the choriocapillaris
vasculature
in a subject in need thereof having AMD, the method comprising administering
to the
subject a therapeutically effective amount of a VE-PTP inhibitor, wherein the
choriocapillaris vasculature is protected; and optionally wherein the subject
is being
treated with anti-VEGF therapy
[0018] Embodiment 12: The method of any one of embodiments 1 through 11,
wherein the choriocapillaris is manipulated in the context of AMD.
[0019] Embodiment 13: The method of any one of embodiments 1 through 11,
wherein the choriocapillaris is manipulated in the context of polypoidal
choroidal
vasculopathy (PCV).
[0020] Embodiment 14: The method of embodiment 13, wherein the PCV is
pachychoroid vasculopathy.
[0021] Embodiment 15: The method of any one of embodiments 1 through 14,
wherein the Tie2 activator is selected from recombinant angiopoietin, BowAng1,
COMP-
Ang1, TSL1, Vasculotide, an Anti-Angiopoietin-2 Binding and Oligomerizing
Antibody,
an Anti-Tie2 Receptor Agonistic Antibody, an Anti-VE-PTP Antibody, a Small
Molecule
VE-PTP Inhibitor, and the like.
8
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0022] Embodiment 16: The method of embodiment 15, wherein the VE-PTP
inhibitor is selected from the small molecule VE-PTP inhibitors as Tie2
activators
described in the specification.
[0023] Embodiment 17: The method of any one of embodiments 1 through 16,
wherein the Tie2 activator and/or the VE-PTP inhibitor are administered to the
eye.
[0024] Embodiment 18: The method of any one of embodiments 1 through 17,
wherein the anti-VEGF therapy is selected from Aflibercept, Bevacizumab,
Pegaptanib
sodium, Ranibizumab, Brolucizumab, Conbercept, Ramucirumab, Faricimab
,Nesvacumab-Aflibercept, Designed ankyrin repeat proteins, Gene-therapy-
targeting
VEGF, and biosimilar versions thereof.
[0025] Embodiment 19: The method of embodiment 11, wherein the anti-VEGF
therapy is administered to the eye.
[0026] Embodiment 20: A mouse carrying at least one angiopoietin 1 null
allele
and at least one wnt1-Cre allele.
[0027] Embodiment 21: The use of a therapeutically effective amount of a
Tie2
activator, optionally in combination with anti-VEGF therapy, for treating AMD
or PCV in
a subject in need thereof.
[0028] Embodiment 22: The use of a therapeutically effective amount of a
Tie2
activator for treating AMD or PCV in a subject in need thereof.
9
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0029] Embodiment 23: The use of a therapeutically effective amount of a
VE-
PTP inhibitor, optionally in combination with anti-VEGF therapy, for treating
AMD or
PCV in a subject in need thereof.
[0030] Embodiment 24: The use of a therapeutically effective amount of a
VE-
PTP inhibitor for treating AMD or PCV in a subject in need thereof.
[0031] Embodiment 25: The use of a therapeutically effective amount of a
Tie2
activator and/or a VE-PTP inhibitor, optionally in combination with anti-VEGF
therapy,
for improving vision in a subject having any one or more of a group of
choriocapillaris
disorders comprising AMD, PCV, variants of choroid polypoid vasculopathy,
geographic
atrophy associated with anti-VEGF therapy, and any other loss of choroid
perfusion
(e.g. diabetic eye disease).
[0032] Embodiment 26: The use of a therapeutically effective amount of a
Tie2
activator and/or a VE-PTP inhibitor for improving vision in a subject having
any one or
more of a group of choriocapillaris disorders comprising AMD, PCV, variants of
choroid
polypoid vasculopathy, geographic atrophy associated with anti-VEGF therapy,
and any
other loss of choroid perfusion (e.g. diabetic eye disease).
[0033] Embodiment 27: The use of a therapeutically effective amount of a
Tie2
activator, optionally in combination with anti-VEGF therapy, for increasing
and/or
improving choriocapillaris vascularization in a subject in need thereof.
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0034] Embodiment 28: The use of a therapeutically effective amount of
a Tie2
activator for increasing and/or improving choriocapillaris vascularization in
a subject in
need thereof.
[0035] Embodiment 29: The use of a therapeutically effective amount of
a VE-
PTP inhibitor, optionally in combination with anti-VEGF therapy, for
increasing and/or
improving choriocapillaris vascularization in a subject in need thereof.
[0036] Embodiment 30: The use of a therapeutically effective amount of
a VE-
PTP inhibitor for increasing and/or improving choriocapillaris vascularization
in a subject
in need thereof.
[0037] Embodiment 31: The use of a therapeutically effective amount of
a Tie2
activator for reducing choriocapillaris dropout during anti-VEGF therapy in a
subject in
need thereof.
[0038] Embodiment 32: The use of a therapeutically effective amount of
a VE-
PTP inhibitor for reducing choriocapillaris dropout during anti-VEGF therapy
in a subject
in need thereof.
[0039] Embodiment 33: The use of a therapeutically effective amount of
a Tie2
activator, optionally in combination with anti-VEGF therapy, for improving
choroidal
perfusion in a subject in need thereof having AMD.
[0040] Embodiment 34: The use of a therapeutically effective amount of
a Tie2
activator for improving choroidal perfusion in a subject in need thereof
having AMD.
= 11
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0041] Embodiment 35: The use of a therapeutically effective amount of a
VE-
PTP inhibitor, optionally in combination with anti-VEGF therapy, for improving
choroidal
perfusion in a subject in need thereof having AMD.
[0042] Embodiment 36: The use of a therapeutically effective amount of a
VE-
PTP inhibitor for improving choroidal perfusion in a subject in need thereof
having AMD.
[0043] Embodiment 37: The use of a therapeutically effective amount of a
Tie2
activator, optionally in combination with anti-VEGF therapy, for protecting
the
choriocapillaris vasculature in a subject in need thereof having AMD.
[0044] Embodiment 38: The use of a therapeutically effective amount of a
Tie2
activator for protecting the choriocapillaris vasculature in a subject in need
thereof
having AMD.
[0045] Embodiment 39: The use of a therapeutically effective amount of a
VE-
PTP inhibitor, optionally in combination with anti-VEGF therapy, for
protecting the
choriocapillaris vasculature in a subject in need thereof having AMD.
[0046] Embodiment 40: The use of a therapeutically effective amount of a
VE-
PTP inhibitor protecting the choriocapillaris vasculature in a subject in need
thereof
having AMD.
[0047] Embodiment 41: The use in any one of embodiments 21 through 40,
wherein the choriocapillaris is manipulated in the context of one or more
choriocapillaris
disorders comprising AMD, PCV, pachychoroid vasculopathy, variants of choroid
12
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
polypoid vasculopathy, geographic atrophy associated with anti-VEGF therapy,
and any
other loss of choroid perfusion (e.g. diabetic eye disease)..
[0048] Embodiment 42: The use in any one of embodiments 21 through 41,
wherein the Tie2 activator is selected from recombinant angiopoietin, BowAng1,
COMP-
Ang1, TSL1, Vasculotide, an Anti-Angiopoietin-2 Binding and Oligomerizing
Antibody,
an Anti-Tie2 Receptor Agonistic Antibody, an Anti-VE-PTP Antibody, a Small
Molecule
VE-PTP Inhibitor, and the like.
[0049] Embodiment 43: The use in any one of embodiments 21 through 41,
wherein the VE-PTP inhibitor is selected from the small molecule VE-PTP
inhibitors as
Tie2 activators described in the specification.
[0050] Embodiment 44: The use in any one of embodiments 21 through 41,
wherein the anti-VEGF therapy is selected from Aflibercept, Bevacizumab,
Pegaptanib
sodium, Ranibizumab, Brolucizumab, Conbercept, Ramucirumab, Faricimab
,Nesvacumab-Aflibercept, Designed ankyrin repeat proteins, Gene-therapy-
targeting
VEGF, and biosimilar versions thereof.
BRIEF DESCRIPTION OF DRAWINGS
[0051] The patent or application file contains at least one drawing
executed in
color. Copies of this patent or patent application publication with color
drawings will be
13
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
provided by the U.S. Patent and Trademark Office upon request and payment of
the
necessary fee.
[0052] FIG. 1: The choriocapillaris is a unique vascular bed which
supplies the
outer retina. Schematic illustration of the human eye highlighting the
location of the
choriocapillaris (CC, red) adjacent to the outer retina. Schlemm's canal, a
lymphatic-like
vessel in the anterior chamber is shown in green. Drawing is not to scale
[0053] FIG. 2A and 2B: ANGPT1 is essential for choriocapillaris
development.
(FIG. 2A) Compared to control littermates, whole-body Angpt1 knockout mice
induced
at E13.5 exhibit sparse EMCN positive capillaries in the peripheral
choriocapillaris (CC)
when imaged at PO. (FIG. 2B) A more severe form of the same phenotype is
observed
in mice lacking Angpt1 in cells of the neural crest lineage, which exhibit
severely
attenuated choriocapillaries when imaged at P1. 20x fields represent an area
of 65,536
pm2, ** p<0.01, *** p <0.001.
[0054] FIG. 3A and 3B: Angiopoietin ligands are highly expressed in the
mouse
choroid. (FIG. 3A) Cryosections from Angpt1GFP mouse eyes reveal robust Angpt1
expression in the choroid as well as in the choroidal melanocytes and retinal
pigmented
epithelium (RPE). Confocal reflectance microscopy (CRM) was used to visualize
these
pigmented tissues. (FIG. 3B) X-Gal staining was used to label Angpt2
expression in the
choroid of adult Angpt2LacZ mice. Angpt2 was observed in the choriocapillaris,
as well
as the choroidal vessels.
14
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0055] FIG. 4: Disorganized EMCN-negative vessels invade the
choriocapillaris
of Angpt1ANC mice by P11. In contrast to the organized pattern of choroidal
vessels
observed in control littermates, a disorganized network of EMCN-negative
vessels was
observed throughout the choriocapillaris of neural crest specific Angpt1
knockout mice
by P11.
[0056] FIG. 5A-5D: VE-PTP is an important regulator of CC development.
(FIG.
5A) Imaged in 5 pm sections or (FIG. 5B) whole-mounts, Ve-ptpNLS-LacZ mice
stained
with anti-Gal antibody reveal strong endothelial Ve-ptp promotor activity in
the
choriocapillaris (CC). (FIG. 5C, quantified in FIG. 5D) Visualized using anti-
endomucin
(EMCN) antibody, whole-body Ve-ptp knockout mice induced at E13.5 have
increased
CC area by PO, indicating an important role for VE-PTP in CC development and
morphogenesis. 20x fields represent an area of 65,536 pm2, * p<0.05, *** p
<0.001
DETAILED DESCRIPTION
DEFINITIONS
[0057] Unless otherwise defined herein, scientific and technical terms
used in
connection with the present disclosure shall have the meanings that are
commonly
understood by those of ordinary skill in the art.
[0058] Unless mentioned otherwise, the techniques employed or
contemplated
herein are standard methodologies well known to one of ordinary skill in the
art. The
practice of the present disclosure will employ, unless otherwise indicated,
conventional
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
techniques of microbiology, tissue culture, molecular biology, chemistry,
biochemistry
and recombinant DNA technology, which are within the skill of the art. The
materials,
methods and examples are illustrative only and not limiting.
[0059] All methods described herein can be performed in any suitable
order
unless otherwise indicated herein or otherwise clearly contradicted by
context. The use
of any and all examples, or exemplary language (e.g. "such as") provided with
respect
to certain embodiments herein is intended merely to better illuminate the
present
disclosure and does not pose a limitation on the scope of the present
disclosure
otherwise claimed. No language in the specification should be construed as
indicating
any non-claimed element as essential to the practice of the present
disclosure.
[0060] Unless specifically stated or obvious from context, as used
herein, the
terms "a", "an", and "the" are understood to be singular or plural.
[0061] 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).
[0062] Unless specifically stated or obvious from context, as used
herein, the
term "about" is understood as within a range of normal tolerance in the art,
for example
16
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
within 2 standard deviations of the mean. About can be understood as within
10%, 9%,
8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated
value.
Unless otherwise clear from context, all numerical values provided herein are
modified
by the term about.
[0063] As used herein, the term "administering" refers to the placement
of a
compound and/or a pharmaceutical composition comprising the compound into a
mammalian tissue or a subject by a method or route that results in at least
partial
localization of the compound and/or composition at a desired site or tissue
location.
[0064] Any compositions, apparatus, systems or methods provided herein
can
be combined with one or more of any of the other compositions, apparatus,
systems or
methods provided herein.
[0065] The terms "disease" or "disorder" are used interchangeably herein
and
refer to any alteration in state of the body or of some of the organs,
interrupting or
disturbing the performance of the functions and/or causing symptoms such as
discomfort, dysfunction, distress, or even death to the person afflicted or
those in
contact with a person. A disease or disorder can also relate to a distemper,
ailing,
ailment, malady, sickness, illness, complaint, indisposition, or affection.
[0066] In this disclosure, the terms "comprise," "have" and "include"
are open-
ended linking verbs. Any forms or tenses of one or more of these verbs, such
as
"comprises," "comprising," "has," "having," "includes" and "including," are
also open-
ended. For example, any method that "comprises," "has" or "includes" one or
more
17
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
steps is not limited to possessing only those one or more steps and can also
cover
other unlisted steps. Similarly, any composition that "comprises," "has" or
"includes" one
or more features is not limited to possessing only those one or more features
and can
cover other unlisted features.
[0067] The terms "consisting essentially of or "consists essentially" of
likewise
has the meaning ascribed in U.S. Patent law and the term is open-ended,
allowing for
the presence of more than that which is recited so long as basic or novel
characteristics
of that which is recited is not changed by the presence of more than that
which is
recited, but excludes prior art embodiments.
[0068] The term "consisting of refers to compositions, methods, and
respective
components thereof as described herein, which are exclusive of any element not
recited
in that description of the embodiment.
[0069] The term "choriocapillaris attenuation" refers to the reduction
of capillary
density in the choroid. It is visible on, for example, flat mount, histologic
sections
through the choroid and by immunohistochemical analysis. It results in loss of
blood
flow and delivery of nutrients to key cells of the choroid including the RPEs.
[0070] The "effectiveness" of a compound or composition of the
disclosure can
be assessed by any method known to one of ordinary skill in the art, including
those
described in the examples of this disclosure. Effectiveness can be established
in vitro
(biochemical and/or biological in cultured cells) and/or in vivo (including in
animal
models). Effectiveness in vitro may be used to extrapolate or predict some
degree of
18
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
effectiveness in vivo, in an animal or in a human subject. A reference or
standard or
comparison may be used. Effective amounts and dosages can be estimated
initially
from in vitro assays. For example, an initial dosage for use in animals can be
formulated
to achieve a circulating blood or serum concentration of active compound that
is at or
above an IC50 of the particular compound as measured in an in vitro assay.
Calculating
dosages to achieve such circulating blood or serum concentrations taking into
account
the bioavailability of the particular compound is well within the capabilities
of skilled
artisans. For guidance, the reader is referred to Fingl & Woodbury, "General
Principles,"
In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1,
pp. 1-
46, latest edition, Pergamagon Press, and the references cited therein, which
methods
are incorporated herein by reference in their entirety. Initial dosages can
also be
estimated from in vivo data, such as animal models. Animal models useful for
testing
the efficacy of compounds to treat or prevent the various diseases described
in this
disclosure are either described herein (see EXAMPLES) or known in the art.
[0071] As used herein, the terms "treatment," "treating," "effective,"
"therapeutically effective" and the like, refer to obtaining a desired
pharmacologic and/or
physiologic effect. The effect can be prophylactic in terms of completely or
partially
preventing a disease or symptom thereof and/or can be therapeutic in terms of
a partial
or complete cure for a disease and/or adverse effect attributable to the
disease.
[0072] By "subject" is meant a mammal, including, but not limited to, a
human or
non-human mammal, such as a bovine, equine, canine, ovine, or feline.
19
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0073] Ranges provided herein are understood to be shorthand for all of
the
values within the range. For example, a range of 1 to 50 is understood to
include any
number, combination of numbers, or sub-range from the group consisting 1, 2,
3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,
26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48,
49, or 50.
EXEMPLARY METHODS OF TREATMENT OF THE DISCLOSURE
[0074] In one embodiment, the disclosure provides a method of treating
age-
related macular degeneration (AMD) (e.g., polypoidal choroidal vasculopathy
(PCV)) in
a subject in need thereof, comprising administering to the subject a
therapeutically
effective amount of a Tie2 activator, optionally in combination with anti-VEGF
therapy.
In an embodiment, the disclosure provides for a use of a therapeutically
effective
amount of a Tie2 activator, optionally in combination with anti-VEGF therapy,
for
treating AMD or PCV in a subject in need thereof. In an embodiment, the
disclosure
provides for a use of a therapeutically effective amount of a Tie2 activator
for treating
AMD or PCV in a subject in need thereof.
[0075] In one embodiment, the disclosure provides a method of treating
AMD or
PCV in a subject in need thereof, comprising administering to the subject a
therapeutically effective amount of a VE-PTP inhibitor, optionally in
combination with
anti-VEGF therapy. In an embodiment, the disclosure provides for a use of a
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
therapeutically effective amount of a VE-PTP inhibitor, optionally in
combination with
anti-VEGF therapy, for treating AMD or PCV in a subject in need thereof. In an
embodiment, the disclosure provides for a use of a therapeutically effective
amount of a
VE-PTP inhibitor for treating AMD or PCV in a subject in need thereof.
[0076] In one embodiment, the disclosure provides a method of improving
vision
in a subject comprising administering to the subject a therapeutically
effective amount of
a Tie2 activator and/or a VE-PTP inhibitor, optionally in combination with
anti-VEGF
therapy, wherein the subject has AMD, choroid polypoid vasculopathy, or
choroid
defects due to complement deficiency or hyperglycemic conditions. In an
embodiment,
the disclosure provides for a use of a therapeutically effective amount of a
Tie2 activator
and/or a VE-PTP inhibitor, optionally in combination with anti-VEGF therapy,
for treating
AMD or PCV in a subject in need thereof. In an embodiment, the disclosure
provides for
a use of a therapeutically effective amount of a Tie2 activator and/or a VE-
PTP inhibitor
for treating AMD or PCV in a subject in need thereof.
[0077] In one embodiment, the disclosure provides a method of increasing
or
improving choriocapillaris vascularization in a subject in need thereof, the
method
comprising administering to the subject a therapeutically effective amount of
a Tie2
activator, optionally in combination with anti-VEGF therapy, wherein
choriocapillaris
vascularization is increased and/or improved in the subject. In an embodiment,
the
21
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
disclosure provides for a use of a therapeutically effective amount of a Tie2
activator,
optionally in combination with anti-VEGF therapy, for increasing and/or
improving
choriocapillaris vascularization in a subject in need thereof. In an
embodiment, the
disclosure provides for a use of a therapeutically effective amount of a Tie2
activator for
increasing and/or improving choriocapillaris vascularization in a subject in
need thereof.
[0078] In one embodiment, the disclosure provides a method of increasing
and/or improving choriocapillaris vascularization in a subject in need
thereof, the
method comprising administering to the subject a therapeutically effective
amount of a
VE-PTP inhibitor optionally in combination with anti-VEGF therapy, wherein
choriocapillaris vascularization is increased and/or improved in the subject.
In an
embodiment, the disclosure provides for a use of a therapeutically effective
amount of a
VE-PTP inhibitor, optionally in combination with anti-VEGF therapy, for
increasing
and/or improving choriocapillaris vascularization in a subject in need
thereof. In an
embodiment, the disclosure provides for a use of a therapeutically effective
amount of a
VE-PTP inhibitor for increasing and/or improving choriocapillaris
vascularization in a
subject in need thereof.
[0079] In one embodiment, the disclosure provides a method of reducing
choriocapillaris dropout during anti-VEGF therapy in a subject in need
thereof, the
method comprising administering to the subject a therapeutically effective
amount of a
Tie2 activator optionally in combination with anti-VEGF therapy, wherein
choriocapillaris
22
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
dropout is reduced. In an embodiment, the disclosure provides for a use of a
therapeutically effective amount of a Tie2 activator, optionally in
combination with anti-
VEGF therapy, for reducing choriocapillaris dropout during anti-VEGF therapy
in a
subject in need thereof.
[0080] In one embodiment, the disclosure provides a method of reducing
choriocapillaris dropout during anti-VEGF therapy in a subject in need
thereof, the
method comprising administering to the subject a therapeutically effective
amount of a
VE-PTP inhibitor optionally in combination with anti-VEGF therapy, wherein
choriocapillaris dropout is reduced. In an embodiment, the disclosure provides
for a use
of a therapeutically effective amount of a VE-PTP inhibitor, optionally in
combination
with anti-VEGF therapy, for reducing choriocapillaris dropout during anti-VEGF
therapy
in a subject in need thereof.
[0081] In one embodiment, the disclosure provides a method of improving
choroidal perfusion in a subject in need thereof having age-related macular
degeneration (AMD), the method comprising administering to the subject a
therapeutically effective amount of a Tie2 activator, wherein choroidal
perfusion is
increased; and optionally wherein the subject is being treated with anti-VEGF
therapy. .
In an embodiment, the disclosure provides for a use of a therapeutically
effective
amount of a Tie2 activator, optionally in combination with anti-VEGF therapy,
for
improving and/or increasing choroidal perfusion in a subject in need thereof.
In an
23
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
embodiment, the disclosure provides for a use of a therapeutically effective
amount of a
Tie2 activator for improving and/or increasing choroidal perfusion in a
subject in need
thereof.
[0082] In one embodiment, the disclosure provides a method of improving
choroidal perfusion in a subject in need thereof having age-related macular
degeneration (AMD), the method comprising administering to the subject a
therapeutically effective amount of a VE-PTP inhibitor, wherein choroidal
perfusion is
increased; and optionally wherein the subject is being treated with anti-VEGF
therapy.
In an embodiment, the disclosure provides for a use of a therapeutically
effective
amount of a VE-PTP inhibitor, optionally in combination with anti-VEGF
therapy, for
improving and/or increasing choroidal perfusion in a subject in need thereof.
In an
embodiment, the disclosure provides for a use of a therapeutically effective
amount of a
VE-PTP inhibitor for improving and/or increasing choroidal perfusion in a
subject in
need thereof.
[0083] In one embodiment, the disclosure provides a method of protecting
the
choriocapillaris vasculature in a subject in need thereof having AMD, the
method
comprising administering to the subject a therapeutically effective amount of
a Tie2
activator, wherein the choriocapillaris vasculature is protected; and
optionally wherein
the subject is being treated with anti-VEGF therapy. In an embodiment, the
disclosure
24
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
provides for a use of a therapeutically effective amount of a Tie2 activator,
optionally in
combination with anti-VEGF therapy, for protecting choriocapillaris
vasculature in a
subject in need thereof. In an embodiment, the disclosure provides for a use
of a
therapeutically effective amount of a Tie2 activator for protecting
choriocapillaris
vasculature in a subject in need thereof.
[0084] In one embodiment, the disclosure provides a method of protecting
the
choriocapillaris vasculature in a subject in need thereof having AMD, the
method
comprising administering to the subject a therapeutically effective amount of
a VE-PTP
inhibitor, wherein the choriocapillaris vasculature is protected; and
optionally wherein
the subject is being treated with anti-VEGF therapy. In an embodiment, the
disclosure
provides for a use of a therapeutically effective amount of a VE-PTP
inhibitor, optionally
in combination with anti-VEGF therapy, for protecting choriocapillaris
vasculature in a
subject in need thereof. In an embodiment, the disclosure provides for a use
of a
therapeutically effective amount of a VE-PTP inhibitor for protecting
choriocapillaris
vasculature in a subject in need thereof.
DISORDERS IN WHICH THE CHOROIDAL VASCULATURE IS IMPAIRED
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0085] The choriocapillaris represents an approximately 10-pm-thin layer
of
relatively large-diameter capillaries interconnected in a densely packed
arrangement
with very small intercapillary pillars located at the inner aspect of the
choroid. The
choriocapillaris is an important vascular layer that is subject to physiologic
changes with
increasing age and that is also associated with a wide range of chorioretinal
diseases,
including age-related macular degeneration (AMD) and central serous
chorioretinopathy
(CSC), which are major causes of vision loss. Diseases or disorders in which
the
choroidal vasculature is impaired (e.g., decreased, functionally impaired,
increased,
damaged, otherwise not normal) are referred to herein as choriocapillaris
disorders. In
one embodiment, the disorder is age-related macular degeneration. In one
embodiment,
the disorder is wet AMD. In one embodiment, the disorder is dry AMD. In one
embodiment, the disorder is polypoidal choroidal vasculopathy (PCV). PCV is
pachychoroid vasculopathy. In one embodiment, the disorder is reduced choroid
perfusion in diabetic eye disease.
[0086] Age-related macular degeneration (AMD) is a leading cause of
central
vision loss worldwide. The progression of dry AMD from early to intermediate
stages is
primarily characterized by increasing drusen formation and adverse impact on
outer
retinal cells. Late stage AMD consists of either geographic atrophy (GA),
which is the
non-exudative (dry) AMD subtype, or choroidal neovascularization, which is the
exudative (wet) AMD subtype. GA is characterized by outer retinal and
choroidal
atrophy, specifically the photoreceptor layer, RPE, and choriocapillaris.
26
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
METHODS OF ASSESSING CHORIOCAPILLARIS VASCULARIZATION
[0087] The choriocapillaris can be visualized in vivo by a variety of
methods
known to one of ordinary skill in the art. In one embodiment, the method
involves either
en face swept-source optical coherence tomography (SS-OCT) or SS-OCT
angiography
(SS-OCTA), which uses decorrelation signal generated from moving erythrocytes.
Both
methods have been shown to have the capability to image the choriocapillaris.
See,
e.g., Wang Q, Chan S, Yang JY, et al. Vascular density in retina and
choriocapillaris as
measured by optical coherence tomography angiography. Am J Ophthalmol. 2016;
168:
95-109.; Zhang Q, Zheng F, Motu!sky EH, et al. A novel strategy for
quantifying
choriocapillaris flow voids using swept-source OCT angiography. Invest
Opthalmology
Vis Sci. 2018; 59: 203-211; and Lane M, Moult EM, Novais EA, et al.
Visualizing the
choriocapillaris under drusen: comparing 1050-nm swept-source versus 840-nm
spectral-domain optical coherence tomography angiography. Invest Opthalmology
Vis
Sci. 2016; 57 (9): 0CT585-0CT590, all of which are incorporated herein by
reference
in their entireties. See also a recent comparison between the methods in Wang,
J.C., et
al. Visualization of Choriocapillaris and Choroidal Vasculature in Healthy
Eyes With En
Face Swept-Source Optical Coherence Tomography Versus Angiography,
Translational
Vision Science & Technology December 2018, Vol.7, 25. doi:10.1167/tvst.7.6.25.
[0088] Various manufacturers offer commercially available OCTA devices.
Currently, the predominant devices on the market are AngioVueTM (Optovue Inc.,
27
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
Fremont, CA, USA), Angioplex and PLEX Elite 9000 (Carl Zeiss Meditec Inc.,
Dublin,
CA, USA), Swept-Source Optical Coherence Tomography AngioTM (Topcon Corp.,
Japan), Heidelberg Spectralis OCTA (Heidelberg, Germany), and Canon OCT-HS100
(Canon, Japan).
[0089] A recent improvement in methods for visualizing the
choriocapillaris has
also been described. Chu, Z. et al. Improving visualization and quantitative
assessment
of choriocapillaris with swept source OCTA through registration and averaging
applicable to clinical systems. Scientific Reports volume 8, Article number:
16826
(2018). The use of these methods in visualizing the choriocapillaris is
another
embodiment of the disclosure.
TIE2 ACTIVATORS
[0090] The following are non-limiting exemplary embodiments of VE-PTP
inhibitors and other Tie2 activators, as well as VEGF inhibitors, that can be
used in the
methods of the disclosure. In certain embodiments, references where the
compounds
are either used or described are provided and are incorporated herein by
reference in
their entireties.
[0091] Recombinant Angiopoietin-1 Proteins as Tie2 Activators
[0092] BowAng1
[0093] Int J Oncol. 2009 Jan;34(1):79-87. Angiopoietin-1/Tie-2
activation
contributes to vascular survival and tumor growth during VEGF blockade. Huang
J, Bae
28
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
JO, Tsai JP, Kadenhe-Chiweshe A, Papa J, Lee A, Zeng S, Komfeld ZN, Ullner P,
Zaghloul N, loffe E, Nandor S, Burova E, Holash J, Thurston G, Rudge J,
Yancopoulos
GD, Yamashiro DJ, Kandel JJ. PMID: 19082480 PMCID: PMC3160826
[0094] Nat Struct Biol. 2003 Jan;10(1):38-44. Angiopoietins have
distinct
modular domains essential for receptor binding, dimerization and
superclustering. Davis
S, Papadopoulos N, Aldrich TH, Maisonpierre PC, Huang T, Kovac L, Xu A,
Leidich R,
Radziejewska E, Rafique A, Goldberg J, Jain V, Bailey K, Karow M, Fandl J,
Samuelsson SJ, loffe E, Rudge JS, Daly TJ, Radziejewski C, Yancopoulos GD.
PMID:
12469114 DOI: 10.1038/nsb880. BowAng1
[0095] COMP-Angl
[0096] Sci Rep. 2015 Oct 19;5:15291. A Designed Angiopoietin-1 Variant,
Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular
Stabilization in N-glycan Dependent Manner.Oh N, Kim K, Kim SJ, Park I, Lee
JE, Seo
YS, An HJ, Kim HM, Koh GY.PMID: 26478188 PMCID: PMC4609988 DOI:
10.1038/srep15291
[0097] Mol Cancer Res. 2009 Dec;7(12):1920-7. Epub 2009 Dec 1.COMP-Ang1
potentiates the antitumor activity of 5-fluorouracil by improving tissue
perfusion in
murine Lewis lung carcinoma.Hwang JA, Lee EH, Kim HW, Park JB, Jeon BH, Cho
CH.PMID: 19952114 DOI: 10.1158/1541-7786.MCR-09-0041.
[0098] Biochem Biophys Res Commun. 2009 Apr 17;381(4):592-6. Epub 2009
Feb 24.COMP-Ang1 ameliorates leukocyte adhesion and reinforces endothelial
tight
29
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
junctions during endotoxemia.Hwang JA, Lee EH, Lee SD, Park JB, Jeon BH, Cho
CH.PMID: 19245790 DOI: 10.1016/j.bbrc.2009.02.096
[0099] Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5553-8. Epub 2004
Apr
1.Designed angiopoietin-1 variant, COMP-Ang1, protects against radiation-
induced
endothelial cell apoptosis.Cho CH, Kammerer RA, Lee HJ, Yasunaga K, Kim KT,
Choi
HH, Kim W, Kim SH, Park SK, Lee GM, Koh GY.PMID: 15060280 PMCID: PMC397421
DOI: 10.1073/pnas.0307575101
[0100] Proc Natl Acad Sci U S A. 2004 Apr 13,101(15):5547-52. Epub 2004
Apr
1.COMP-Ang1: a designed angiopoietin-1 variant with nonleaky angiogenic
activity.Cho
CH, Kammerer RA, Lee HJ, Steinmetz MO, Ryu YS, Lee SH, Yasunaga K, Kim KT, Kim
I, Choi HH, Kim W, Kim SH, Park SK, Lee GM, Koh GY.PMID: 15060279 PMCID:
PMC397420 001: 10.1073/pnas.0307574101
[0101] Synthetic Angiopoietin-1 Mimetic Ligands as Tie2 Activators
[0102] TSL1
[0103] Mol Pharm. 2018 Sep 4;15(9):3962-3968. Epub 2018 Aug 6.
[0104] Development of an Orthogonal Tie2 Ligand Resistant to Inhibition
by
Ang2.Issa E, Moss AJ, Fischer M, Kang M, Ahmed S, Farah H, Bate N, Giakomidi
D,
Brindle NP.PMID: 30036484 DOI: 10.1021/acs.molpharmaceut.8b00409.
[0105] Vasculotide
[0106] Cell Transplant. 2018 Dec;27(12):1744-1752. Epub 2018 Aug
20.Angiopoietin-1 Mimetic Peptide Promotes Neuroprotection after Stroke in
Type 1
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
Diabetic Rats.Venkat P, Yan T, Chopp M, Zacharek A, Ning R, Van Slyke P,
Dumont D,
Landschoot-Ward J, Liang L, Chen J.PMID: 30124060 PMCID: PMC6300775 DOI:
10.1177/0963689718791568
[0107] Br J Anaesth. 2018 Nov;121(5):1041-1051. Epub 2018 Jun
19.Vasculotide, an angiopoietin-1 mimetic, reduces pulmonary vascular leakage
and
preserves microcirculatory perfusion during cardiopulmonary bypass in rats.
Dekker
NAM, van Meurs M, van Leeuwen ALI, Hofland HM, van Slyke P, Vonk ABA, Boer C,
van den Brom CE.PMID: 30336848 DOI: 10.1016/j.bja.2018.05.049
[0108] Anesthesiology. 2018 Feb;128(2):361-374. Vasculotide, an
Angiopoietin-
1 Mimetic, Restores Microcirculatory Perfusion and Microvascular Leakage and
Decreases Fluid Resuscitation Requirements in Hemorrhagic Shock.Trieu M, van
Meurs M, van Leeuwen ALI, Van Slyke P, Hoang V, Geeraedts LMG Jr, Boer C, van
den Brom CE.PMID: 28968277 001: 10.1097/ALN.0000000000001907
[0109] Crit Care. 2017 Nov 13;21(1):274. Vasculotide reduces pulmonary
hyperpermeability in experimental pneumococcal pneumonia.Gutbier B, Jiang X,
Dietert
K, Ehrler C, Lienau J, Van Slyke P, Kim H, Hoang VC, Maynes JT, Dumont DJ,
Gruber
AD, Weissmann N, Mitchell TJ, Suttorp N, Witzenrath M.PMID: 29132435 PMCID:
PMC5683375 DOI: 10.1186/s13054-017-1851-6
[0110] BMC Res Notes. 2016 May 28;9:289.Vasculotide, an Angiopoietin-1
mimetic, ameliorates several features of experimental atopic dermatitis-like
31
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
disease.Bourdeau A, Van Slyke P, Kim H, Cruz M, Smith T, Dumont DJ.PMID:
27236199 PMCID: PMC4884390 DOI: 10.1186/s13104-015-1817-1
[0111] Sci Rep. 2016 Feb 25;6:22111. The Synthetic Tie2 Agonist Peptide
Vasculotide Protects Renal Vascular Barrier Function In Experimental Acute
Kidney
Injury.RUbig E, Stypmann J, Van Slyke P, Dumont DJ, Spieker T, Buscher K,
Reuter S,
Goerge T, Pavenstadt H, Kumpers P.PMID: 26911791 PMCID: PMC4766468 DOI:
10.1038/srep22111
[0112] Sci Rep. 2015 Jun 5;5:11030. The Tie2-agonist Vasculotide rescues
mice from influenza virus infection.Sugiyama MG, Armstrong SM, Wang C, Hwang
D,
Leong-Poi H, Advani A, Advani S, Zhang H, Szaszi K, Tabuchi A, Kuebler WM, Van
Slyke P, Dumont DJ, Lee WL.PMID: 26046800 PMCID: PMC4457136 DOI:
10.1038/srep11030
[0113] EMBO Mol Med. 2015 Jun;7(6):770-87. Vasculotide reduces
endothelial
permeability and tumor cell extravasation in the absence of binding to or
agonistic
activation of Tie2.Wu FT, Lee CR, Bogdanovic E, Prodeus A, Gariepy J, Kerbel
RS.PMID: 25851538 PMCID: PMC4459817 DOI: 10.15252/emmm.201404193
[0114] BMC Cancer. 2014 Aug 26,14:614.Vasculotide, an Angiopoietin-1
mimetic, reduces acute skin ionizing radiation damage in a preclinical mouse
model.Korpela E, Yohan D, Chin LC, Kim A, Huang X, Sade S, Van Slyke P, Dumont
DJ, Liu SK.PMID: 25159192 PMCID: PMC4159535 DOI: 10.1186/1471-2407-14-614
32
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
[0115] Tissue Eng Part A. 2009 Jun;15(6):1269-80.Acceleration of
diabetic
wound healing by an angiopoietin peptide mimetic.Van Slyke P, Alami J, Martin
D,
Kuliszewski M, Leong-Poi H, Sefton MV, Dumont D.PMID: 18939935 DOI:
10.1089/ten.tea.2007.0400
[0116] Anti-Angiopoietin-2 Binding and Oligomerizing Antibody as Tie2
Activator
[0117] Any anti-angiopoietin 2 antibody can be used with the methods of
the
disclosure and one of ordinary skill in the art would be aware of the same.
Non-limiting
examples include those described inSci Adv. 2019 Feb 13;5(2):eaau6732.Tie2
activation promotes choriocapillary regeneration for alleviating neovascular
age-related
macular degeneration.Kim J, Park JR, Choi J, Park!, Hwang Y, Bae H, Kim Y,
Choi W,
Yang JM, Han S, Chung TY, Kim P, Kubota Y, Augustin HG, Oh WY, Koh GY.PMID:
30788433 PMCID: PMC6374104 DOI: 10.1126/sciadv.aau6732
[0118] J Clin Invest. 2017 Oct 2;127(10):3877-3896.Impaired
angiopoietinfrie2
signaling compromises Schlemm's canal integrity and induces glaucoma.Kim J,
Park
DY, Bae H, Park DY, Kim D, Lee CK, Song S, Chung TY, Lim DH, Kubota Y, Hong
YK,
He Y, Augustin HG, Oliver G, Koh GYPMID: 28920924 PMCID: PMC5617682 DOI:
10.1172/JCI94668
[0119] Cancer Cell. 2016 Dec 12;30(6):953-967. Normalization of Tumor
Vessels by Tie2 Activation and Ang2 Inhibition Enhances Drug Delivery and
Produces a
Favorable Tumor Microenvironment.Park JS, Kim IK, Han S, Park 1, Kim C, Bae J,
Oh
33
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
SJ, Lee S, Kim JH, Woo DC, He Y, Augustin HG, Kim 1, Lee D, Koh GY.PMID:
27960088 DOI: 10.1016/j.cce11.2016.10.018
[0120] Sci Transl Med. 2016 Apr 20;8(335):335ra55.Amelioration of sepsis
by
TIE2 activation-induced vascular protection.Han S, Lee SJ, Kim KE, Lee HS, Oh
N,
Park I, Ko E, Oh SJ, Lee YS, Kim D, Lee S, Lee DH, Lee KH, Chae SY, Lee JH,
Kim
SJ, Kim HC, Kim S, Kim SH, Kim C, Nakaoka Y, He Y, Augustin HG, Hu J, Song PH,
Kim YI, Kim P, Kim I, Koh GY.PMID: 27099174 DOI: 10.1126/scitranslmed.aad9260
[0121] Anti-Tie2 Receptor Agonistic Antibody as Tie2 Activator
[0122] Any anti-Tie2 antibody can be used with the methods of the
disclosure
and one of ordinary skill in the art would be aware of the same. Non-limiting
examples
include those described inBionnaterials. 2015 May;51:119-128.Stimulation of
angiogenesis and survival of endothelial cells by human monoclonal Tie2
receptor
antibody. Hwang B, Lee SH, Kim JS, Moon JH, Jeung IC, Lee NG, Park J, Hong HJ,
Cho YL, Jung H, Park YJ, Lee SJ, Lee HG, Kim WK, Han BS, Bae KH, Chung SJ,
Kwon
YG, Lee SC, Kim SJ and Min JK. PMID: 25771003 DOI:
10.1016/j.biomaterials.2015.01.062.
VE-PTP INHIBITORS.
[0123] Anti-VE-PTP Antibody as Tie2 Activator
[0124] Any antibody against VE-PTP can be used with the methods of the
disclosure and one of ordinary skill in the art would be aware of the same.
Non-limiting
34
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
examples include those described inJ Exp Med. 2015 Dec 14;212(13):2267-87.
Epub
2015 Dec 7.Interfering with VE-PTP stabilizes endothelial junctions in vivo
via Tie-2 in
the absence of VE-cadherin Frye M, Dierkes M, Kiippers V, Vockel M, Tomm J,
Zeuschner D, Rossaint J, Zarbock A, Koh GY, Peters K, Nottebaum AF, Vestweber
D.PMID: 26642851 PMCID: PMC4689167 DOI: 10.1084/jem.20150718
[0125] J Clin Invest. 2014 Oct;124(10):4564-76. Epub 2014 Sep
2.Targeting
VE-PTP activates TIE2 and stabilizes the ocular vasculature.Shen J, Frye M,
Lee BL,
Reinardy JL, McClung JM, Ding K, Kojima M, Xia H, Seidel C, Lima e Silva R,
Dong A,
Hackett SF, Wang J, Howard BW, Vestweber D, Kontos CD, Peters KG, Campochiaro
PA.PMID: 25180601 PMCID: PMC4191011 DOI: 10.1172/JCI74527
[0126] J Cell Biol. 2009 May 18;185(4):657-71. VE-PTP controls blood
vessel
development by balancing Tie-2 activity.Winderlich M, Keller L, Cagna G,
Broermann A,
Kamenyeva 0, Kiefer F, Deutsch U, Nottebaum AF, Vestweber D.PMID: 19451274
PMCID: PMC2711575 DOI: 10.1083/jcb.200811159
[0127] Small Molecule VE-PTP Inhibitors as Tie2 Activators
[0128] Any small molecule agent that inhibits VE-PTP can be used with
the
methods of the disclosure and one of ordinary skill in the art would be aware
of the
same. Non-limiting examples include those described in: Bioorg Chem. 2018
Dec;81:270-277. Epub 2018 Jun 6. Zhang W, Wei Z, et al.; Microsurgery. 2017
Sep;37(6):624-631. Epub 2016 Nov 17.Zor F, Meric C et al.;J Infect Dis. 2017
Mar
1;215(5):813-817.0ehlers SH et al. ; Curr Diab Rep. 2016 Dec;16(12):126.
Review.
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
Campochiaro PA et al.; Ophthalmology. 2016 Aug;123(8):1722-1730. Epub 2016 May
26Campochiaro PA et al.; Acta Neuropathol. 2016 May;131(5):753-73. Epub 2016
Mar
1.Gurnik S, et al.; J Exp Med. 2015 Dec 14;212(13):2267-87. Epub 2015 Dec
7Frye M,
et al.; Ophthalmology. 2015 Mar;122(3):545-54. Epub 2014 Nov 12.Campochiaro PA
et
al.; J Clin Invest. 2014 Oct;124(10):4564-76. Epub 2014 Sep 2.; Shen J et
al.Targeting
VE-PTP activates TIE2 and stabilizes the ocular vasculature. et al.; Antioxid
Redox
Signal. 2014 May 10;20(14):2130-40. Epub 2014 Feb 4.Zeng LF et al; J Natl
Cancer
Inst. 2013 Aug 21;105(16):1188-201. Epub 2013 Jul 30. Goel S et al.;J Biol
Chem. 2012
Mar 16;287(12):9322-6. Epub 2012 Jan 24.Wilson M et al.; Angiogenesis.
2009;12(1):25-33. Epub 2009 Jan 1.Yacyshyn OK et al.; Acta Crystallogr D Biol
Crystallogr. 2006 Dec;62(Pt 12):1435-45. Epub 2006 Nov 23. Evdokinnov AG et
al.;
Bioorg Med Chem Lett. 2006 Aug 15;16(16):4252-6. Epub 2006 Jun 12.Amarasinghe
KK et al.; Proc Natl Acad Sci U S A. 2006 Jul 11;103(28):10606-11. Epub 2006
Jun
29.Ntiren-Miiller A et al.: Am J Physiol Heart Circ Physiol. 2004
Jul;287(1):H268-76.
Epub 2004 Feb 26. Carr AN et al.; J Biol Chem. 2004 Jun 4;279(23):24226-35.
Epub
2004 Mar 15. ; Lund IK, et al.; J lnorg Biochem. 2003 Aug 1;96(2-3):321-30.;
Peters KG
et at.
ANTI-VEGF TREATMENTS.
[0129] Any agent that inhibits VEGF can be used with the methods of the
disclosure and one of ordinary skill in the art would be aware of the same.
Non-limiting
36
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
examples include: Affibercept, trade name Eylea and Zaltrap, is a recombinant
fusion
protein.; Bevacizumab, trade name Avastin, is a humanized recombinant
monoclonal
IgG1 antibody.; Pegaptanib sodium, trade name Macugen, is a pegylated
synthetic RNA
aptamer.; Ranibizumab, trade name Lucentis, is a humanized IgG1 monoclonal
antibody fragment.; Brolucizumab is a humanized single chain antibody
fragment;
Conbercept is a fusion protein that acts as a VEGF receptor decoy; Ramucirumab
is a
novel human IgG1 monoclonal antibody that selectively inhibits the VEGFR2 and
blocks
the VEGFR2-related signaling and activating pathways; Faricimab (RG7716), is a
bispecific anti-VEGF/ANG2 monoclonal antibody capable of binding,
neutralizing, and
depleting VEGF-A and ANG-2.; Nesvacumab-Affibercept is Ang-2NEGF-A co-
formulation of two monoclonal antibodies.; Designed ankyrin repeat proteins
(DARPins)
are various formulations of genetically engineered small recombinant non-
immunoglobulin proteins that mimic antibodies in their ability to bind
specific proteins
with high affinity and specificity; biosimilars of all of the listed
antibodies; Gene-therapy-
targeting VEGF; small molecules, etc.
METHODS OF ASSESSING CHORIOCAPILLARIS PERFUSION
[0130] One of ordinary skill in the art would be familiar with methods
to assess
choriocapillaris perfusion. In one embodiment, choriocapillaris perfusion is
assessed
37
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
through the use of passive fluorescent dyes. In one embodiment,
choriocapillaris
perfusion is assessed through fluorescent dye angiography.
METHODS OF ASSESSING PROTECTION OF THE CHORIOCAPILLARIS
One of ordinary skill in the art would be familiar with methods to assess
protection of the
choriocapillaris. In one embodiment, the choriocapillaris is visualized as
described
above in METHODS OF ASSESSING CHORIOCAPILLARIS VASCULARIZATION. In
one embodiment, the choriocapillaris is visualized prior to treatment, at
different stages
during treatment, and at different stages after a treatment that damages the
choriocapillaris. In one embodiment, the treatment is anti-VEGF treatment. In
one
embodiment, a Tie2 activator is administered in combination with the anti-VEGF
treatment in order to protect the choriocapillaris from anti-VEGF treatment-
induced/related damage. Exemplary embodiments of different combination
treatment
regimens are exemplified below.
METHODS OF ASSESSING CHORIOCAPILLARIS DROPOUT
[0131] The term "choriocapillaris dropout" refers to the loss of
functional
capillaries supplying blood to the choroid (e.g. could be physical loss due to
rarefaction
or it could be to non-functional vessels). Methods for assessing this
parameter are
known in the art and described elsewhere in this application.
38
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
METHODS OF ASSESSING IMPROVEMENTS IN VISION
[0132] In one embodiment, improvements in vision are measured by a
subject's
uncorrected distance and near visual acuity, which may be taken using a
standard
acceptable eye chart.
[0133] In another embodiment, improvements in vision are measured by
clinical
evaluation of the depth of field, which may be obtained either using standard
wavefront
aberrometry or according to other methods well known to one of ordinary skill
in the art.
[0134] In yet another embodiment, improvements in vision are measured by
change of pupil size, which may be evaluated by the infrared imaging system
used for
checking alignment during auto-refractometry. The pupil size can also be
measured by
Aberronneter and pupilometer.
[0135] In yet another embodiment, improvements in vision are measured by
pupil appearance, which can include inspecting the pupils for equal size (1 mm
or less
of difference may be normal), regular shape, reactivity to light, and direct
and
consensual accommodation.
[0136] In yet another embodiment, improvements in vision are measured by
non-invasive objective assessments of the 3rd, 4th and 5th ocular higher order
39
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
aberrations (such as coma, spherical aberration, and trefoil), which may be
conducted
using standard wavefront aberrometry.
METHODS OF ADMINISTRATION
[0137] The Tie2 activators, VE-PTP inhibitors, and anti-VEGF treatments,
can
be administered in unit forms of administration to mammalian subjects,
including human
beings. Suitable unit forms of administration include, as non-limiting
examples, forms
administered orally and forms administered via a parenteral/systemic route,
non-limiting
examples of which including inhalation, subcutaneous administration,
intramuscular
administration, intravenous administration, intradermal administration,
intravitreal
administration, as well as topical and local ocular (i.e, subconjunctival,
intravitreal,
retrobulbar, intracameral) modes of administration. In one embodiment, the
treatment is
injected into the eye (intraocular injection). Can we add in nanoparticle -
mediated
delivery?
[0138] In some embodiments, pharmaceutical compositions for parenteral
administration can be in the form of aqueous solutions, non¨aqueous solutions,
suspensions, emulsions, drops (including, as a non-limiting example, eye
drops), or any
combination(s) thereof. In some embodiments, such pharmaceutical compositions
may
comprise one or more of water, pharmaceutically acceptable glycol(s),
pharmaceutically
acceptable oil(s), pharmaceutically acceptable organic esters, or other
pharmaceutically
acceptable solvents. A variety of vehicles suitable for administering
compounds to the
eye are known in the art. Specific non-limiting examples are described in U.S.
Pat. No.
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
6,261,547; U.S. Pat. No. 6,197,934; U.S. Pat. No. 6,056,950; U.S. Pat. No.
5,800,807;
U.S. Pat. No. 5,776,445; U.S. Pat. No. 5,698,219; U.S. Pat. No. 5,521,222;
U.S. Pat.
No. 5,403,841; U.S. Pat. No. 5,077,033; U.S. Pat. No. 4,882,150; and U.S. Pat.
No.
4,738,851, all of which are incorporate herein by reference in their entirety.
[0139] With regard to these treatments, the mode (or modes) of
administration,
dosage (or dosages), and optimized pharmaceutical form (or forms) can be
determined
according to criteria generally considered during the establishment of a
treatment of a
patient, such as, by way of non-limiting examples, the potency of the
compound(s)
and/or pharmaceutically acceptable salts of the compound(s), the age of the
patient, the
body weight of the patient, the severity of the patient's condition (or
conditions), the
patient's tolerance to the treatment, and secondary effects observed in
treatment.
Determination of dosages effective to provide therapeutic benefit for specific
modes and
frequency of administration is within the capabilities of those skilled in the
art.
COMBINATION TREATMENTS
[0140] In one embodiment, the disclosure provides combination treatments
for
use in the treatment of choriocapillaris disorders such as, for example, AMD.
[0141] Each therapeutic agent in a combination therapy of the invention
may be
administered simultaneously (i.e., in the same medicament), concurrently
(i.e., in
separate medicaments administered one right after the other in any order) or
sequentially in any order. Sequential administration is particularly useful
when the
41
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
therapeutic agents in the combination therapy are in different dosage forms
(one agent
is a tablet or capsule and another agent is a sterile liquid) and/or are
administered on
different dosing schedules, e.g., a Tie2 activator that is administered at
least daily and a
anti-VEGF treatment that is administered less frequently, such as once weekly,
once
every two weeks, or once every three weeks.
[0142] In some embodiments, at least one of the therapeutic agents in
the
combination therapy is administered using the same dosage regimen (dose,
frequency
and duration of treatment) that is typically employed when the agent is used
as
monotherapy for treating the same disease or another disease. In other
embodiments,
the patient receives a lower total amount of at least one of the therapeutic
agents in the
combination therapy than when the agent is used as monotherapy, e.g., smaller
doses,
less frequent doses, and/or shorter treatment duration.
[0143] In some embodiments, a combination therapy of the invention is
administered to a patient who has not been previously treated with either one
or both of
the individual treatments, i.e., is treatment-naïve. In other embodiments, the
combination therapy is administered to a patient who failed to achieve a
sustained
response after prior therapy with one of the agents, i.e., is treatment-
experienced.
[0144] In one embodiment, the Tie2 activator and/or the VE-PTP
inhibitors are
administered together with the anti-VEGF treatment to a treatment-naïve
patient. In
another embodiment, the Tie2 activator and/or the VE-PTP inhibitors are
administered
42
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
after the patient has started the anti-VEGF treatment and the treatment is
either not
sufficient or there are side effects (e.g., choriocapillaris dropout).
[0145] Selecting a dosage regimen (also referred to herein as an
administration
regimen) for a combination therapy of the invention depends on several
factors,
including the serum or tissue turnover rate of the entity, the level of
symptoms, the
immunogenicity of the entity, and the accessibility of the target cells,
tissue or organ in
the subject being treated. Optionally, a dosage regimen maximizes the amount
of each
therapeutic agent delivered to the patient consistent with an acceptable level
of side
effects. Accordingly, the dose amount and dosing frequency of each
biotherapeutic and
chemotherapeutic agent in the combination depends in part on the particular
therapeutic
agent, the severity of the choriocapillaris disorder being treated, and
patient
characteristics. Determination of the appropriate dosage regimen may be also
made by
the clinician, e.g., using parameters or factors known or suspected in the art
to affect
treatment or predicted to affect treatment, and will depend, for example, the
patient's
clinical history (e.g., previous therapy), and the severity of the
choriocapillaris disorder.
[0146] Regardless of the treatment indication, there are essentially two
regimens for administering anti-VEGF drugs: continuous and intermittent/as
required (or
pro re nata, PRN for short).
[0147] The following non-limiting examples and data illustrate various
aspects
and features relating to the methods and uses of compounds of the present
invention. In
some embodiments, the present methods and uses of compounds provide results
and
43
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
data which are surprising, unexpected and contrary thereto. While the utility
of this
invention is illustrated through the use of several compounds and
moieties/groups which
can be used therewith, it will be understood by those skilled in the art that
comparable
results are obtainable with various other compounds, moieties and/or groups,
as are
commensurate with the scope of this invention.
EXAMPLES
EXAMPLE 1
Angiopoietin-Tie2 signaling is a key regulator of choriocapillaris development
[0148] Data presented herein indicate, for the first time, that Angpt-
Tie2
signaling is essential throughout choroid development and that mice lacking
Angptl
from the entire body or only in their neural-crest derived cells including
melanocytes of
the choroid, exhibit dramatic choriocapillaris attenuation. This term refers
to a reduction
of capillary density in the choroid. It is visible on flat mount, histologic
sections through
the choroid and by immunohistochemical analysis. It results in loss of blood
flow and
delivery of nutrients to key cells of the choroid including the RPEs. Indeed,
experiments
using whole-body Angptl knockout mice deleted at E13.5, after the first stage
of
choriocapillaris development, revealed sparse choriocapillaris by postnatal
day (P) 0
(FIG. 2 A), highlighting the critical role of Angpt-Tie2 signaling in late-
stage
development. Importantly, given the expression pattern of Angptl (FIG. 3 A) in
choroidal melanocytes, and the fact that melanocytes have recently been
reported as
an important regulator of choriocapillaris development, we developed a mouse
deficient
44
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
in Angptl in melanocytes. Importantly, the mechanisms and growth factors
produced
by choroidal melanocytes needed for choroid development have not been
identified.
The data suggest that Angptl may be one such¨if not the¨melanocyte-secreted
factor responsible for this regulation. To investigate this possibility, and
to understand
the role of Angptl in early choroid development, a novel neural-crest specific
Angptl
knockout mouse was generated using WntlCre (53, 54). Angptl ANC mice lack
Angptl
in the choroidal melanocytes and other non-endothelial cells of the choroid
and sclera
but retain expression in the RPE. Compared to littermate controls, AngptlANC
mice
show a markedly attenuated choriocapillary network at P1 (FIG. 2 B),
confirming the
importance of choroidal Angptl.
EXAMPLE 2
Deficits in Angpt-TIE2 signaling lead to choriocapillaris dysfunction,
including capillary
rarefaction and PCV-like disorganized vascular networks
[0149] Angptl knockout mice induced at E13.5, after completion of
initial
choriocapillaris development (16), exhibit reduced choriocapillary density
(FIG. 2A),
suggesting that Angpt-Tie2 signaling is essential for late-stage
choriocapillaris
development, growth or maintenance. Using neural crest-specific Angptl
knockout
mice, it was discovered that by P11, while the capillary sparsity seen at P1
was still
observed, a branched network of large, disorganized vessels had formed in the
choriocapillaris of mutant mice, suggesting a continued role for Angpt
signaling after
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
birth (FIG. 4). These abnormal vessels were positive for the endothelial
marker PODXL,
but EMCN negative, suggesting either loss of the choriocapillary phenotype or
encroachment of intermediate or large choroidal vessels into the
choriocapillaris. This
finding is strikingly similar to the disorganized networks commonly observed
by
indocyanine green angiography in eyes with polypoidal choroidal vasculopathy
(PCV), a
little understood group of adult-onset AMD-like diseases including
pachychoroid
vasculopathy (62) which are characterized by a disorganized network of dilated
choroidal vessels and polypoid lesions (24-26).
[0150] The data suggest that Angpt-TIE2 signaling plays an important
role in
choriocapillaris homeostasis and deletion of Angpt ligands or TIE2 leads to
dysfunction.
Identification of PCV-like vascular networks in AngptlANC mice provides an
important
proof-of-concept, and characterization of these mice provides mechanistic
insights on
the unusual encroachment of large disorganized vessels into the
choriocapillaris¨a key
aspect of this poorly understood disease.
EXAMPLE 3
VE-PTP deletion leads to TIE2 activation and choriocapillary growth, and will
protect the
choriocapillaris during geographic atrophy and anti-VEGF therapy
[0151] VE-PTP is an endothelial receptor-type phosphatase which
specifically
dephosphorylates TIE2, reducing its signaling activity (41, 48). Loss of VE-
PTP activity,
either by genetic deletion or pharmacological inhibition, results in increased
TIE2 activity
46
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
in vivo even in the absence of ANGPT1, leading to elevated signaling through
PI3K/AKT
and eNOS ((41). Results presented herein indicate that Ve-ptp is strongly
expressed by
the adult mouse choriocapillaris (FIG. 5 A,B), highlighting its role in the
choriocapillary
endothelium. In addition to regulating vascular stability, TIE2 activation can
cause
venous widening (47, 68) and data presented herein suggest that embryonic
deletion of
Ve-ptp at E13.5 leads to increased choriocapillaris vascular density by PO
(FIG. 5 C,D).
Thus, the results indicate that Ve-ptp is strongly expressed in the adult
mouse
choriocapillaris, and that knockout mice have dramatically increased
choriocapillary
area. Importantly, we have recently demonstrated that elevated VE-PTP
expression can
worsen micro-vascular disease outcomes. Renal VE-PTP expression is upregulated
by
hypertension and hyperglycemia (Carota et al. JEM 2019, in press), leading to
reduced
TIE2 signaling and enhancing a cycle of microvascular dysfunction in diabetes.
VE-PTP
knockout mice have dramatically increased choriocapillary area, suggesting
that this
protein could provide an effective target for therapy aimed at increasing
choriocapillaris
function in early AMD or preventing capillary dropout during anti-VEGF
therapy.
[0152] As
choriocapillaris rarefaction is a component of AMD, this suggests that
VE-PTP inhibition may provide an effective pro-choriocapillaris therapy. In
vivo, VE-PTP
inhibition leads to increased activation of PI3K/AKT and ENOS signaling, pro-
survival
pathways which are also activated by VEGF. Therefore, VE-PTP inhibition as
combination therapy may protect the choriocapillaris during treatment with
anti-VEGF
47
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
agents, preventing choriocapillaris rarefaction and limiting potential side
effects of anti-
VEGF treatment.
EXAMPLE 4
Generation of the Mice Used in the Examples
[0153] Angptl, Angpt2 and Tie2 knockout mice have been previously
described
(39, 40, 55). As global deletion of Angptl before E13.5 is incompatible with
life, specific
deletion in the periocular mesenchyme is essential for study of the
choriocapillaris
during early embryogenesis. While periocular endothelial cells arise from the
mesoderm, non-endothelial cells and melanocytes are neural crest-derived (54,
56).
Therefore, the well-characterized WntlCre line (53, 54) was used to achieve
deletion of
Angptl in the neural crest lineage.
[0154] A novel neural-crest specific Angptl knockout mouse using Wnt/Cre
(53, 54) was generated. AngptlANC mice lack Angptl in the choroidal
melanocytes and
other non-endothelial cells of the choroid and sclera but retain expression in
the RPE.
Compared to littermate controls, Angpti ANC mice show a markedly attenuated
choriocapillary network at P1 (FIG. 2 B), confirming the importance of
choroidal Angptl.
These mice are a good model of for AMD and for Polypoidal choroidal
vasculopathy,
which is an AMD-like disease that predominantly affects patients of African-
American
and Asian descent and has no existing animal model.
[0155] Inducible VE-PTP knockout mice (Vete"; Rosa261frA; TetOnCre) are
described in Carota et al. JEM 2019, published.
48
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
INCORPORATION BY REFERENCE
[0156] Various publications, articles and patents are cited or described
in the
background and throughout the specification; each of these references is
herein
incorporated by reference in its entirety. Discussion of documents, acts,
materials,
devices, articles or the like which has been included in the present
specification is for
the purpose of providing context for the invention. Such discussion is not an
admission
that any or all of these matters form part of the prior art with respect to
any inventions
disclosed or claimed.
[0157] REFERENCES CITED AND OTHER SUPPORTING REFERENCES.
1. Bird AC. Therapeutic targets in age-related macular disease. J Clin
Invest.
2010;120(9):3033-41.
2. Wong WL, Su X, Li X, Cheung CMG, Klein R, Cheng C-Y, and Wong TY. Global
prevalence of age-related macular degeneration and disease burden projection
for 2020
and 2040: A systematic review and meta-analysis. The Lancet Global Health.
2014;2(2):e106-e16.
3. Stefansson E, GeirsdOttir A, and Sigurdsson H. Metabolic physiology in
age related
macular degeneration. Prog Retin Eye Res. 2011;30(1):72-80.
4. Dietzel M, Pauleikhoff D, Holz FG, and Bird AC. In: Holz FG, Pauleikhoff
D, Spaide RF,
and Bird AC eds. Age-related macular degeneration. Berlin, Heidelberg:
Springer Berlin
Heidelberg; 2013:101-9.
5. Pauleikhoff D, Spital G, Radermacher M, Brumm GA, Lommatzsch A, and Bird
AC. A
fluorescein and indocyanine green angiographic study of choriocapillaris in
age-related
macular disease. Arch Ophthalmol. 1999;117(10):1353-8.
6. Lutty G, Grunwald J, Majji AB, Uyama M, and Yoneya S. Changes in
choriocapillaris and
retinal pigment epithelium in age-related macular degeneration. Mol Vis.
1999;5(35.
7. Metelitsina TI, Grunwald JE, Dupont JC, Ying G-S, Brucker AJ, and
Dunaief JL.
Foveolar choroidal circulation and choroidal neovascularization in age-related
macular
degeneration. Invest Ophthalmol Vis ScL 2008;49(1):358-63.
8. Fleckenstein M, Schmitz-Valckenberg S, Sunness JS, and Holz FG. In: Holz
FG,
Pauleikhoff D, Spaide RF, and Bird AC eds. Age-related macular degeneration.
Berlin,
Heidelberg: Springer Berlin Heidelberg; 2013:121-38.
9. Spilsbury K, Garrett KL, Shen W-Y, Constable IJ, and Rakoczy PE.
Overexpression of
vascular endothelial growth factor (vegf) in the retinal pigment epithelium
leads to the
49
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
development of choroidal neovascularization. The American Journal of
Pathology.
2000;157(1):135-44.
10. Inoue Y, Yanagi Y, Matsuura K, Takahashi H, Tamaki Y, and Araie M.
Expression of
hypoxia-inducible factor 1a and 2a in choroidal neovascular membranes
associated with
age-related macular degeneration. Br J Ophthalmol. 2007;91(12):1720-1.
11. Sheridan CM, Pate S, Hiscott P, Wong D, Pattwell DM, and Kent D.
Expression of
hypoxia-inducible factor-1a and -2a in human choroidal neovascular membranes.
Graefe's Archive for Clinical and Experimental Ophthalmology.
2009;247(10):1361-7.
12. Mendrinos E, and Pournaras CJ. Topographic variation of the choroidal
watershed zone
and its relationship to neovascularization in patients with age-related
macular
degeneration. Acta Ophthalmol (Copenh). 2009;87(3):290-6.
13. Mcleod DS, Grebe R, Bhutto I, Merges C, Baba T, and Lutty GA.
Relationship between
rpe and choriocapillaris in age-related macular degeneration. Invest
Ophthalmol Vis
2009;50(10):4982-91.
14. Seddon JM, Mcleod D, Bhutto IA, and Et Al. Histopathological insights
into choroidal
vascular loss in clinically documented cases of age-related macular
degeneration. JAMA
Ophthalmology. 2016;134(11):1272-80.
15. Young M, Chui L, Fallah N, Or C, Merkur AB, Kirker AW, Albiani DA, and
Forooghian F.
Exacerbation of choroidal and retinal pigment epithelial atrophy after anti-
vascular
endothelial growth factor treatment in neovascular age-related macular
degeneration.
Retina. 2014;34(7):1308-15.
16. Marneros AG, Fan J, Yokoyama Y, Gerber HP, Ferrara N, Crouch RK, and
Olsen BR.
Vascular endothelial growth factor expression in the retinal pigment
epithelium is
essential for choriocapillaris development and visual function. The American
Journal of
Pathology.167(5):1451-9.
17. Peters S, Heiduschka P, Julien S, Ziemssen F, Fietz H, Bartz-Schmidt
KU, and
Schraermeyer U. Ultrastructural findings in the primate eye after intravitreal
injection of
bevacizumab. Am J Ophtha/mo/.143(6):995-1002.e2.
18. Shimomura Y, Hirata A, Ishikawa S, and Okinami S. Changes in
choriocapillaris
fenestration of rat eyes after intravitreal bevacizumab injection. Graefe's
Archive for
Clinical and Experimental Ophthalmology. 2009;247(8):1089-94.
19. Kurihara T, Westenskow PD, Bravo S, Aguilar E, and Friedlander M.
Targeted deletion
of vegfa in adult mice induces vision loss. J Clin Invest. 2012;122(11):4213-
7.
20. Grunwald JE, Daniel E, Huang J, Ying G-S, Maguire MG, Toth CA, Jaffe
GJ, Fine SL,
Blodi B, Klein ML, et al. Risk of geographic atrophy in the comparison of age-
related
macular degeneration treatments trials. Ophthalmology.121(1):150-61.
21. Chakravarthy U, Harding SP, Rogers CA, Downes SM, Lotery AJ, Culliford
LA, and
Reeves BC. Alternative treatments to inhibit vegf in age-related choroidal
neovascularisation: 2-year findings of the ivan randomised controlled trial.
The
Lancet.382(9900):1258-67.
22. Sho K, Takahashi K, Yamada H, Wada M, Nagai Y, Otsuji T, Nishikawa M,
Mitsuma Y,
Yamazaki Y, Matsumura M, et al. Polypoidal choroidal vasculopathy: Incidence,
demographic features, and clinical characteristics. Arch Ophthalmol.
2003;121(10):1392-
6.
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
23. Yannuzzi LA, Sorenson J, Spaide RF, and Lipson B. Idiopathic polypoidal
choroidal
vasculopathy (ipcv). Retina. 1990;10(1):1-8.
24. Nakajima M, Yuzawa M, Shimada H, and Mori R. Correlation between
indocyanine
green angiographic findings and histopathology of polypoidal choroidal
vasculopathy.
Jpn J Ophthalmol. 2004;48(3):249-55.
25. Laude A, Cackett PD, Vithana EN, Yeo IY, Wong D, Koh AH, Wong TY, and
Aung T.
Polypoidal choroidal vasculopathy and neovascular age-related macular
degeneration:
Same or different disease? Prog Retin Eye Res. 2010;29(1):19-29.
26. Yuzawa M, Mori R, and Kawamura A. The origins of polypoidal choroidal
vasculopathy.
Br J Ophthalmol. 2005;89(5):602-7.
27. Hatz K, and Prunte C. Polypoidal choroidal vasculopathy in caucasian
patients with
presumed neovascular age-related macular degeneration and poor ranibizumab
response. Br J Ophthalmol. 2014;98(2):188-94.
28. Stangos AN, Gandhi JS, Nair-Sahni J, Heimann H, Pournaras CJ, and
Harding SP.
Polypoidal choroidal vasculopathy masquerading as neovascular age-related
macular
degeneration refractory to ranibizumab. Am J Ophthalmol. 2010;150(5):666-73.
29. Kawashima Y, Oishi A, Tsujikawa A, Yamashiro K, Miyake M, Ueda-Arakawa
N,
Yoshikawa M, Takahashi A, and Yoshimura N. Effects of aflibercept for
ranibizumab-
resistant neovascular age-related macular degeneration and polypoidal
choroidal
vasculopathy. Graefe's Archive for Clinical and Experimental Ophthalmology.
2015;253(9):1471-7.
30. Koh A, Lee WK, Chen L-J, Chen S-J, Hashad Y, Kim H, Lai TY, Pilz S,
Ruamviboonsuk
P, Tokaji E, et al. Everest study: Efficacy and safety of verteporfin
photodynamic therapy
in combination with ranibizumab or alone versus ranibizumab monotherapy in
patients
with symptomatic macular polypoidal choroidal vasculopathy. Retina.
2012;32(8):1453-
64.
31. Tan CS, Ngo WK, Chen JP, Tan NW, and Lim TH. Everest study report 2:
Imaging and
grading protocol, and baseline characteristics of a randomised controlled
trial of
polypoidal choroidal vasculopathy. Br J Ophthalmol. 2015;99(5):624-8.
32. Koh A, Lai TYY, Takahashi K, Wong TY, Chen L-J, Ruamviboonsuk P, Tan
CS, Feller C,
Margaron P, Lim TH, et al. Efficacy and safety of ranibizumab with or without
verteporfin
photodynamic therapy for polypoidal choroidal vasculopathy: A randomized
clinical trial.
JAMA ophthalmology. 2017;135(11):1206-13.
33. Ma L, Brelen ME, Tsujikawa M, Chen H, Chu WK, Lai TYY, Ng DSC, Sayanagi
K, Hara
C, Hashida N, et al. Identification of angpt2 as a new gene for neovascular
age-related
macular degeneration and polypoidal choroidal vasculopathy in the chinese and
japanese populationsangpt2 in amd and pcv. Invest Ophthalmol Vis Sci.
2017;58(2):1076-83.
34. Lutty GA, Hasegawa T, Baba T, Grebe R, Bhutto I, and Mcleod DS.
Development of the
human choriocapillaris. Eye. 2010;24(408.
35. Hasegawa T, Mcleod DS, Bhutto IA, Prow T, Merges CA, Grebe R, and Lutty
GA. The
embryonic human choriocapillaris develops by hemo-vasculogenesis. Dev Dyn.
2007;236(8):2089-100.
51
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
36. Saint-Geniez M, Kurihara T, Sekiyama E, Maldonado AE, and D'amore PA.
An essential
role for rpe-derived soluble vegf in the maintenance of the choriocapillaris.
Proceedings
of the National Academy of Sciences. 2009;106(44):18751-6.
37. Augustin HG, Young Koh G, Thurston G, and Alitalo K. Control of
vascular
morphogenesis and homeostasis through the angiopoietin-tie system. Nat Rev Mol
Cell
Biol. 2009;10(3):165-77.
38. Kim J, Park JR, Choi J, Park I, Hwang Y, Bae H, Kim Y, Choi W, Yang JM,
Han S, et al.
Tie2 activation promotes choriocapillary regeneration for alleviating
neovascular age-
related macular degeneration. Science Advances. 2019;5(2):eaau6732.
39. Gale NW, Thurston G, Hackett SF, Renard R, Wang Q, Mcclain J, Martin C,
Witte C,
Witte MH, Jackson D, et al. Angiopoietin-2 is required for postnatal
angiogenesis and
lymphatic patterning, and only the latter role is rescued by angiopoietin-1.
Dev Cell.
2002;3(3):411-23.
40. Davis S, Aldrich TH, Jones PF, Acheson A, Compton DL, Jain V, Ryan TE,
Bruno J,
Radziejewski C, Maisonpierre PC, et al. Isolation of angiopoietin-1, a ligand
for the tie2
receptor, by secretion-trap expression cloning. Cell. 1996;87(7):1161-9.
41. Souma T, Thomson BR, Heinen S, Carota IA, Yamaguchi S, Onay T, Liu P,
Ghosh AK,
Li C, Eremina V, et al. Context-dependent functions of angiopoietin 2 are
determined by
the endothelial phosphatase veptp. Proceedings of the National Academy of
Sciences.
2018.
42. Thomson BR, Heinen S, Jeansson M, Ghosh AK, Fatima A, Sung H-K, Onay T,
Chen H,
Yamaguchi S, Economides AN, et al. A lymphatic defect causes ocular
hypertension and
glaucoma in mice. J Clin Invest. 2014;124(10).
43. Souma T, Tompson SW, Thomson BR, Siggs OM, Kizhatil K, Yamaguchi S,
Feng L,
Limviphuvadh V, Whisenhunt KN, Maurer-Stroh S, et al. Angiopoietin receptor
tek
mutations underlie primary congenital glaucoma with variable expressivity. J
Clin
Invest.126(7):2575-87.
44. Thomson BR, Souma T, Tompson SW, Onay T, Kizhatil K, Siggs OM, Feng L,
Whisenhunt KN, Yanovitch TL, Kalaydjieva L, et al. Angiopoietin-1 is required
for
schlemm's canal development in mice and humans. J Clin Invest.
2017;127(12):4421-
36.
45. Nambu H, Nambu R, Oshima Y, Hackett SF, Okoye G, Wiegand S, Yancopoulos
G,
Zack DJ, and Campochiaro PA. Angiopoietin 1 inhibits ocular neovascularization
and
breakdown of the blood¨retinal barrier. Gene Ther. 2004;11(865.
46. Lee J, Park D-Y, Park DY, Park I, Chang W, Nakaoka Y, Komuro I, Yoo 0-
J, and Koh
GY. Angiopoietin-1 suppresses choroidal neovascularization and vascular
leakage.
Invest Ophthalmol Vis ScL 2014;55(4):2191-9.
47. Thurston G, Wang Q, Baffert F, Rudge J, Papadopoulos N, Jean-Guillaume
D, Wiegand
S, Yancopoulos GD, and Mcdonald DM. Angiopoietin 1 causes vessel enlargement,
without angiogenic sprouting, during a critical developmental period.
Development.
2005;132(14):3317.
48. Winderlich M, Keller L, Cagna G, Broermann A, Kamenyeva 0, Kiefer F,
Deutsch U,
Nottebaum AF, and Vestweber D. Ve-ptp controls blood vessel development by
balancing tie-2 activity. The Journal of Cell Biology. 2009;185(4):657-71.
52
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
49. Shen J, Frye M, Lee BL, Reinardy JL, Mcclung JM, Ding K, Kojima M, Xia
H, Seidel C,
Silva RLE, et al. Targeting ve-ptp activates t1e2 and stabilizes the ocular
vasculature. J
Clin Invest. 2014;124(10):4564-76.
50. Campochiaro PA, Khanani A, Singer M, Patel S, Boyer D, Dugel P, Kherani
S, Withers
B, Gambino L, Peters K, et al. Enhanced benefit in diabetic macular edema from
akb-
9778 tie2 activation combined with vascular endothelial growth factor
suppression.
Ophthalmology. 2016;123(8):1722-30.
51. Kenig-Kozlovsky Y, Scott RP, Onay T, Carota IA, Thomson BR, Gil HJ,
Ramirez V,
Yamaguchi S, Tanna CE, Heinen S, et al. Ascending vasa recta are
angiopoietin/tie2-
dependent lymphatic-like vessels. J Am Soc Nephrol. 2017.
52. Shibuya H, Watanabe R, Maeno A, Ichimura K, Tamura M, Wakana S,
Shiroishi T, Ohba
K, Takeda K, Tomita H, et al. Melanocytes contribute to the vasculature of the
choroid.
Genes Genet Syst. 2018;advpub(
53. Snedeker J, Schock EN, Struve JN, Chang OF, Cionni M, Tran PV, Brugmann
SA, and
Stottmann RW. Unique spatiotemporal requirements for intraflagellar transport
genes
during forebrain development. PLoS One. 2017;12(3):e0173258.
54. Gage PJ, Rhoades W, Prucka SK, and Hjalt T. Fate maps of neural crest
and mesoderm
in the mammalian eye. Invest Ophthalmol Vis ScL 2005;46(11):4200-8.
55. Dumont DJ, Gradwohl G, Fong GH, Puri MC, Gertsenstein M, Auerbach A,
and
Breitman ML. Dominant-negative and targeted null mutations in the endothelial
receptor
tyrosine kinase, tek, reveal a critical role in vasculogenesis of the embryo.
Genes Dev.
1994;8(16):1897-909.
56. Etchevers HC, Vincent C, Le Douarin NM, and Couly GF. The cephalic
neural crest
provides pericytes and smooth muscle cells to all blood vessels of the face
and
forebrain. Development. 2001;128(7):1059-68.
57. Zhou BO, Ding L, and Morrison SJ. Hematopoietic stem and progenitor
cells regulate the
regeneration of their niche by secreting angiopoietin-1. Elife. 2015;4(e05521.
58. Kim J, Park D-Y, Bae H, Park DY, Kim D, Lee C-K, Song S, Chung T-Y, Lim
DH, Kubota
Y, et al. Impaired angiopoietin/tie2 signaling compromises schlemm's canal
integrity and
induces glaucoma. J Clin Invest. 2017;127(10):3877-96.
59. Chu M, Li T, Shen B, Cao X, Zhong H, Zhang L, Zhou F, Ma W, Jiang H,
and Xie P.
Angiopoietin receptor tie2 is required for vein specification and maintenance
via
regulating coup-tfii. Elife. 2016;5(e21032.
60. Baba T, Grebe R, Hasegawa T, Bhutto I, Merges C, Mcleod DS, and Lutty
GA.
Maturation of the fetal human choriocapillaris. Invest Ophthalmol Vis Sci.
2009;50(7):3503-11.
61. Chu M, Li T, Shen B, Cao X, Zhong H, Zhang L, Zhou F, Ma W, Jiang H,
Xie P, et al.
Angiopoietin receptor tie2 is required for vein specification and maintenance
via
regulating coup-thi. eLife. 2016;5(e21032.
62. Akkaya S. Spectrum of pachychoroid diseases. Int Ophthalmol.
2018;38(5):2239-46.
63. Jeansson M, Gawlik A, Anderson G, Li C, Kerjaschki D, Henkelman M, and
Quaggin SE.
Angiopoietin-1 is essential in mouse vasculature during development and in
response to
injury. J Clin Invest. 2011;121(6):2278-89.
53
CA 3075146 2020-03-11
PATENT
Attorney Docket No.: 14002.6004-00000
64. Thomson Br. CI, Souma T., Kizhatil K., Feng L., Liu X., John Sw., Young
TI., Quaggin.,
Se. Defects in angiopoietin-tie2 signaling lead to dose-dependent glaucoma in
mice
ARVO. Seattle, WA; May 5, 2016, 2016.
65. Mcmenamin PG. Optimal methods for preparation and immunostaining of
iris, ciliary
body, and choroidal wholemounts. Invest Ophthalmol Vis ScL 2000;41(10):3043-8.
66. Terasaki H, Miyake Y, Suzuki T, Nakamura M, and Nagasaka T. Polypoidal
choroidal
vasculopathy treated with macular translocation: Clinical pathological
correlation. Br J
Ophthalmol. 2002;86(3):321-7.
67. Thurston G, Rudge JS, loffe E, Zhou H, Ross L, Croll SD, Glazer N,
Holash J, Mcdonald
DM, and Yancopoulos GD. Angiopoietin-1 protects the adult vasculature against
plasma
leakage. Nat Med. 2000;6(4):460-3.
68. Limaye N, Wouters V, Uebelhoer M, Tuominen M, Wirkkala R, Mulliken JB,
Eklund L,
Boon LM, and Vikkula M. Somatic mutations in angiopoietin receptor gene tek
cause
solitary and multiple sporadic venous malformations. Nat Genet. 2009;41(11:118-
24.
69. Bhutto IA, Ogura S, Baldeosingh R, Mcleod DS, Lutty GA, and Edwards MM.
An acute
injury model for the phenotypic characteristics of geographic atrophy. Invest
Ophthalmol
Vis Sci. 2018;59(4):AMD143-AMD51.
70. Keir LS, Firth R, Aponik L, Feitelberg D, Sakimoto S, Aguilar E, Welsh
GI, Richards A,
Usui Y, Satchell SC, et al. Vegf regulates local inhibitory complement
proteins in the eye
and kidney. J Clin Invest. 2017;127(11:199-214.
71. Mullins RF, Dewald AD, Streb LM, Wang K, Kuehn MH, and Stone EM.
Elevated
membrane attack complex in human choroid with high risk complement factor h
genotypes. Exp Eye Res. 2011;93(4):565-7.
72. Clark SJ, Perveen R, Hakobyan S, Morgan BP, Sim RB, Bishop PN, and Day
AJ.
Impaired binding of the age-related macular degeneration-associated complement
factor
h 402h allotype to bruch's membrane in human retina. The Journal of Biological
Chemistry. 2010;285(39):30192-202.
73. Kannan R, and Hinton DR. Sodium iodate induced retinal degeneration:
New insights
from an old model. Neural regeneration research. 2014;9(23):2044-5.
74. Zieger M, and Punzo C. Improved cell metabolism prolongs photoreceptor
survival upon
retinal-pigmented epithelium loss in the sodium iodate induced model of
geographic
atrophy. Oncotarget. 2016;7(9):9620-33.
54
CA 3075146 2020-03-11
