U.S. patent application number 16/740143 was filed with the patent office on 2020-07-16 for methods of treatment of keloid using an anti-vegf agent.
The applicant listed for this patent is Randal Tanh Hoang Pham. Invention is credited to Randal Tanh Hoang Pham.
Application Number | 20200222533 16/740143 |
Document ID | / |
Family ID | 50845370 |
Filed Date | 2020-07-16 |
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United States Patent
Application |
20200222533 |
Kind Code |
A1 |
Pham; Randal Tanh Hoang |
July 16, 2020 |
Methods Of Treatment Of Keloid Using An Anti-VEGF Agent
Abstract
Methods for treating pterygium recurrence following
pterygiectomy, and for treating keloid recurrence, following
surgical removal of the keloid, are disclosed. The methods include
administering an anti-VEGF agent (e.g., antibody (e.g.,
bevacizumab) or small molecule inhibitor of VEGF signaling), or a
combination therapy that includes co-administering an anti-VEGF
agent, with an anti-inflammatory steroid and/or a non-steroidal
anti-inflammatory drug (NSAID) to a subject.
Inventors: |
Pham; Randal Tanh Hoang;
(San Jose, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Pham; Randal Tanh Hoang |
San Jose |
CA |
US |
|
|
Family ID: |
50845370 |
Appl. No.: |
16/740143 |
Filed: |
January 10, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15700992 |
Sep 11, 2017 |
10556008 |
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16740143 |
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14861943 |
Sep 22, 2015 |
9757452 |
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15700992 |
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14261198 |
Apr 24, 2014 |
9161978 |
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14861943 |
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13830819 |
Mar 14, 2013 |
8747852 |
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14261198 |
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61746778 |
Dec 28, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 39/3955 20130101;
A61K 31/573 20130101; C07K 16/22 20130101; A61K 31/196 20130101;
A61K 31/58 20130101; A61K 2039/505 20130101; A61P 29/00 20180101;
A61K 45/06 20130101; A61K 31/573 20130101; A61K 2300/00 20130101;
A61K 31/196 20130101; A61K 2300/00 20130101; A61K 39/3955 20130101;
A61K 2300/00 20130101 |
International
Class: |
A61K 39/395 20060101
A61K039/395; A61K 31/196 20060101 A61K031/196; A61K 31/573 20060101
A61K031/573; A61K 45/06 20060101 A61K045/06; C07K 16/22 20060101
C07K016/22; A61K 31/58 20060101 A61K031/58 |
Claims
1. A method for treating keloid recurrence, the method comprising
administering (1) an anti-VEGF agent to a site where a keloid has
been removed from a subject.
2. The method of claim 1, wherein the anti-VEGF agent is an
antibody or small molecule inhibitor of VEGF signaling.
3. The method of claim 1, wherein the anti-VEGF agent is the
antibody bevacizumab.
4. The method according to claim 3, wherein the bevacizumab is
administered at a dose of 10 mg.
5. The method of claim 1, wherein the anti-VEGF agent is
administered by injection or topically.
6. The method of claim 5, wherein the anti-VEGF agent is injected
at or adjacent to a site of new blood vessel growth at the site of
keloid removal.
7. The method of claim 1, further comprising administering to the
site (2) an anti-inflammatory steroid or (3) a non-steroidal
anti-inflammatory drug (NSAID), or both.
8. The method of claim 1, wherein the anti-inflammatory steroid is
applied topically, systemically, or by injection to the site of
keloid removal.
9. The method of claim 1, wherein the anti-inflammatory steroid is
alclometasone, diflorasone, fluocinonide, or prednicarbate.
10. The method of claim 1, wherein the NSAID is diclofenac.
11. The method of claim 1, further comprising administering to the
site one or more subsequent administrations of one or more of an
anti-VEGF agent, an anti-inflammatory steroid, and an NSAID.
Description
RELATED APPLICATIONS
[0001] The present application is a continuation of U.S.
application Ser. No. 15/700,992, filed Sep. 11, 2017, which is a
continuation of U.S. application Ser. No. 14/861,943, filed Sep.
22, 2015, now U.S. Pat. No. 9,757,452, which is a division of U.S.
application Ser. No. 14/261,198, filed Apr. 24, 2014, now U.S. Pat.
No. 9,161,978, which is a continuation of U.S. application Ser. No.
13/830,819, filed Mar. 14, 2013, now U.S. Pat. No. 8,747,852, which
claims the benefit of U.S. Provisional Patent Application Ser. No.
61/746,778, filed Dec. 28, 2012. The foregoing prior applications
are herein incorporated by reference in their entireties.
TECHNICAL FIELD
[0002] The present disclosure relates to methods of treating eye
and skin conditions, in particular, methods for treating pterygium
and keloid, including treating recurrence of these conditions
following surgery.
BACKGROUND
[0003] Pterygium is a fibrovascular conjunctiva proliferation of
the supra-ocular surface with extension to the cornea. Surgery is
the accepted method for primary treatment of pterygium; however,
one of the major limitations of surgical management of pterygium is
the high recurrence rate. The growth factor, vascular endothelial
growth factor (VEGF), has been shown to be highly expressed in
pterygium and in the growing blood vessels that are associated with
pterygium recurrence.
[0004] Bevacizumab (Avastin.RTM.) is a 149-Kd full length humanized
anti-vascular endothelial growth factor (anti-VEGF) antibody that
has been used off-label as a therapy for ocular neovascular
diseases, such as age-related macular degeneration. Others have
also found that subconjunctival administration of bevacizumab
provided temporary treatment of recurrent pterygium.
[0005] Keloid is a chronic skin condition that results in formation
of tumor-like growths on the skin. Despite its benign nature,
keloid can cause severe aesthetic and, in some cases, functional
problems which negatively impacts a person's quality of life. Like
pterygia, keloid tissue expresses excessive amounts of VEGF, which
may play a role in the formation and evolution of keloid.
SUMMARY OF THE INVENTION
[0006] In certain aspects, the present disclosure provides a method
for treating pterygium recurrence comprising administering to an
affected eye of a subject in need of such treatment (1) an
anti-VEGF agent; and one or both of (2) an anti-inflammatory
steroid and (3) a non-steroidal anti-inflammatory drug (NSAID). In
certain aspects, the method comprises administering (1) an
anti-VEGF agent and (2) an anti-inflammatory steroid to the
affected eye of the subject. In certain aspects, the treatment
comprises administering (1) an anti-VEGF agent and (3) an NSAID to
the affected eye of the subject. In some aspects, the treatment
comprises administering (1) an anti-VEGF agent, (2) an
anti-inflammatory steroid and (3) an NSAID to the affected eye of
the subject. In certain aspects, the anti-VEGF agent is an antibody
or small molecule inhibitor of VEGF signaling. In other aspects,
the anti-VEGF agent is an antibody. In some aspects, the anti-VEGF
agent is bevacizumab. In certain aspects, the amount of bevacizumab
administered is about 2.5 mg. In other aspects, the anti-VEGF agent
is injected at a site of new blood vessel growth in the affected
eye. In other aspects, the anti-VEGF agent is administered by
injection. In some aspects, the anti-inflammatory steroid is
prednisolone acetate. In certain aspects, the anti-inflammatory
steroid is prednisolone acetate 1%. In other aspects, the
anti-inflammatory steroid is difluprednate. In certain aspects, the
anti-inflammatory steroid is difluprednate 0.05%. In some aspects,
the anti-inflammatory steroid is triamcinolone acetonide. In
certain aspects, the amount of triamcinolone acetonide administered
is 40 mg. In still other aspects, the anti-inflammatory steroid is
in the form of an ophthalmic solution or suspension. In certain
aspects the anti-inflammatory steroid is administered topically. In
still other aspects, the anti-inflammatory steroid is injected. In
some aspects, the anti-inflammatory steroid is co-injected with the
anti-VEGF agent. In other aspects, the NSAID is selected from the
group consisting of diclofenac, ketorolac, bromfenac and nepafenac.
In certain aspects, the NSAID is in the form of an ophthalmic
solution or suspension. In other aspects, the NSAID is selected
from the group consisting of diclofenac 0.1%, ketorolac
tromethamine ophthalmic solution 0.4%, bromfenac ophthalmic
solution 0.09%, and nepafenac ophthalmic suspension 0.1%. In some
aspects, the NSAID is administered topically.
[0007] In some aspects, the above treatments are performed in
conjunction with surgery to remove pterygium. In certain aspects,
the treatment is commenced within 2 weeks prior to or following the
surgery to remove pterygium. In other aspects, the treatment is
commenced within 1 week prior to or following the surgery to remove
pterygium. In certain aspects, the treatment is commenced following
surgery to remove pterygium. In other aspects, the treatment is
commenced within 2 weeks following the surgery to remove pterygium.
In other aspects, the treatment is commenced within 1 week
following the surgery to remove pterygium. In certain aspects, the
commencement of the treatment comprises administering at least one
of (1) the anti-VEGF agent; (2) the anti-inflammatory steroid and
(3) the non-steroidal anti-inflammatory drug (NSAID). In certain
aspects, the treatment comprises more than one administration of
the anti-VEGF agent to the affected eye of the subject. In other
aspects, the treatment comprises more than one administration of
the anti-inflammatory steroid or the NSAID or both to the affected
eye of the subject. In certain aspects, the anti-VEGF agent is
administered at least once before the anti-inflammatory steroid or
the NSAID or both are administered. In other aspects, the
anti-inflammatory steroid or the NSAID is administered to the
affected eye of the subject at least once before the anti-VEGF
agent is administered. In some aspects, the anti-inflammatory
steroid and the NSAID are each administered to the affected eye of
the subject at least once before the anti-VEGF agent is
administered. In other aspects, the anti-VEGF agent, the
anti-inflammatory steroid and the NSAID are each administered on
separate days at least once. In still other aspects, the anti-VEGF
agent and the anti-inflammatory steroid are administered at least
once on the same day. In still other aspects, the subject has had
pterygium recurrence prior to commencement of the treatment. In
certain aspects, the subject is human.
[0008] In another aspect, the present disclosure provides a method
for treating keloid recurrence, the method comprising administering
(1) an anti-VEGF agent to a site where a keloid has been removed
from a subject. In certain aspects, the anti-VEGF agent is an
antibody or small molecule inhibitor of VEGF signaling. In other
aspects, the anti-VEGF agent is an antibody. In some aspects, the
anti-VEGF agent is bevacizumab. In certain aspects, the bevacizumab
is administered at a dose of 10 mg. In other aspects, the anti-VEGF
agent is administered by injection or topically. In certain
aspects, the anti-VEGF agent is injected at or adjacent to a site
of new blood vessel growth at the site of keloid removal. In some
aspects, the treatment further comprises administering to the site
(2) an anti-inflammatory steroid or (3) a non-steroidal
anti-inflammatory drug (NSAID), or both. In certain aspects, the
treatment further comprises administering (1) an anti-VEGF agent,
(2) an anti-inflammatory steroid and (3) a non-steroidal
anti-inflammatory drug (NSAID) to the site. In certain aspects, the
anti-inflammatory steroid is co-administered with the anti-VEGF
agent. In other aspects, the anti-inflammatory steroid is applied
topically, systemically, or by injection to the site of keloid
removal. In certain aspects, the anti-inflammatory steroid is
alclometasone, diflorasone, fluocinonide, or prednicarbate. In some
aspects, the anti-inflammatory steroid is alclometasone 0.005%
cream, diflorasone 0.005% cream, fluocinonide cream 0.1%, or
prednicarbate topical 0.1% cream or ointment. In certain aspects,
the NSAID is diclofenac. In some aspects, the NSAID is diclofenac
1% solution. In other aspects, the treatment further comprises
administering to the site one or more subsequent administrations of
the anti-VEGF agent. In some aspects, the treatment further
comprises administering to the site one or more subsequent
administrations of one or more of an anti-VEGF agent, an
anti-inflammatory steroid, and an NSAID. In certain aspects, the
subject is human.
[0009] Unless otherwise defined, 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 pertains. In
case of conflict, the present document, including definitions
provided herein, will control.
[0010] All publications, patent applications, patents, GenBankx
Accession numbers, protocols and other references mentioned herein
are incorporated by reference in their entirety for all purposes.
The materials, methods, and examples disclosed herein are
illustrative only and not intended to be limiting.
[0011] The details of one or more embodiments of the invention are
set forth in the accompanying drawings and the description below.
Preferred methods and materials are described below, although
methods and materials similar or equivalent to those described
herein can also be used in the practice or testing of the present
invention. Other features, objects, and advantages of the invention
will be apparent from the description and drawings, and from the
claims.
DESCRIPTION OF DRAWINGS
[0012] FIGS. 1-5 are timelines showing the treatment regimens of
Patients 1-5.
DETAILED DESCRIPTION
[0013] The present disclosure provides methods for treating
pterygium recurrence. Pterygia are caused by a neovascular
condition of the eye that can eventually lead to blindness. In
certain aspects, the methods disclosed herein include administering
a combination therapy to an affected eye of a subject with
recurrent pterygia. First, a pterygium is removed (e.g.,
surgically) from an affected eye, and then, following removal, the
patient is treated with a combination therapy. The combination
therapy disclosed herein includes treating the affected eye with an
anti-VEGF agent (such as, e.g., the antibody, bevacizumab, or other
inhibitor of VEGF signaling) and with an anti-inflammatory steroid
(e.g., triamcinolone acetonide (injected), or prednisolone acetate
1% or difluprednate 0.05% (topical)) and/or a non-steroidal
anti-inflammatory drug (NSAID) (e.g., diclofenac 0.1%, ketorolac
tromethamine ophthalmic solution, 0.4%, bromfenac ophthalmic
solution, 0.09%, nepafenac ophthalmic suspension 0.1%). As
disclosed herein, the anti-inflammatory steroid and/or NSAID can be
administered before or after the eye is treated with the anti-VEGF
agent.
[0014] Since it has been reported in the literature that treatment
of recurrent pterygia using the anti-VEGF antibody bevacizumab was
not effective or only temporarily effective (e.g., 6 months or
less), it was surprising to discover, as disclosed herein, that
bevacizumab, when administered as part of a combination therapy,
was effective for inhibiting pterygium recurrence. For example, as
shown in FIGS. 1, 2, 3A, 3B, 4, and 5, patients receiving a
combination therapy including administration of bevacizumab,
steroid and NSAID, have not had recurrence of pterygium in more
than 11 months, 8 months, 6 months, 9 months, and 3 months,
respectively.
[0015] The present disclosure also provides methods for treating
keloid recurrence. The methods disclosed herein include
administering to the site where a keloid has been removed an
anti-VEGF agent, such as, but not limited to an anti-VEGF antibody,
e.g., bevacizumab. While not intending to be limited to any one
particular theory or mechanism of action, keloids express high
levels of VEGF, and thus the administration of anti-VEGF antibody
is believed to inhibit neovascularization, which is required for
keloid growth. The methods disclosed herein also include
administering the anti-VEGF agent in a combination therapy with one
or both of an anti-inflammatory steroid (e.g., alclometasone 0.005%
cream, diflorasone 0.005% cream, fluocinonide cream 0.1%, or
prednicarbate topical 0.1% cream or ointment) and an NSAID (e.g.,
diclofenac 1% solution).
Definitions
[0016] As used herein, the term "pterygium recurrence" means the
reappearance of pterygium in the eye following removal (e.g.,
surgically) of primary pterygium.
[0017] As used herein, the term "subject" means any mammal, and, in
particular, a human, and can also be referred to, e.g., as an
individual or patient. A "subject in need of treatment" pterygium
or keloid recurrence according to the methods disclosed herein is a
subject who is at risk of pterygium recurrence or keloid
recurrence, e.g., a patient who is about to undergo or has recently
(e.g., within about 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1
month(s), about 2 or 1 week(s) or less) undergone, or is about to
undergo (e.g., within about 1 month, about 1 week or less) surgery
to remove a pterygium or keloid.
[0018] As used herein, an "anti-VEGF agent" means an inhibitor of
VEGF signaling. Anti-VEGF agents include antibodies (e.g.,
bevacizumab), antibody fragments (e.g., an antibody light chain
(VL), an antibody heavy chain (VH), a single chain antibody (scFv),
an F(ab')2 fragment, a Fab fragment, an Fd fragment, an Fv
fragment, and a single domain antibody fragment (DAb). Fragments
can be obtained, e.g., via chemical or enzymatic treatment of an
intact or complete antibody or antibody chain or by recombinant
means), fusion proteins, peptide, nucleic acids (e.g., siRNA,
shRNA), and other small molecules, etc. that disrupt the
interaction between VEGF (VEGF-A) and its receptor
(VEGFR-1/VEGFR-2). Other, non-limiting examples of anti-VEGF agents
encompassed by the present disclosure are provided herein
below.
[0019] As used herein, the term "adjacent to", e.g., in the context
of injecting an anti-VEGF adjacent to or near the site of new blood
vessel growth, means proximate to (e.g., within about 0.1 mm, 0.2
mm, 0.3 mm, 0.4 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, 4 mm, or 5 mm from
the site of blood vessel growth).
[0020] As used herein the terms "therapeutically effective" and
"effective amount", used interchangeably, applied to a dose or
amount refer to a quantity of a composition, compound or
pharmaceutical formulation that is sufficient to result in a
desired activity upon administration to a subject in need thereof.
Within the context of the present invention, the term
"therapeutically effective" refers to that quantity of a
composition, compound or pharmaceutical formulation that is
sufficient to reduce, eliminate or delay at least one symptom of a
disease or condition specified herein, e.g., pterygium or keloid
recurrence. When a combination of active agents is administered,
the effective amount of the combination, or individual agents, may
or may not include amounts of each agent that would have been
effective if administered individually. The dosage of the
therapeutic formulation will vary, depending upon the nature of the
disease or condition, the patient's medical history, the frequency
of administration, the manner of administration, the clearance of
the agent from the host, and the like. The initial dose may be
larger, followed by smaller maintenance doses. The dose may be
administered, e.g., weekly, biweekly, daily, semi-weekly, etc., to
maintain an effective dosage level.
[0021] Therapeutically effective dosages in the methods described
herein can be determined by the treating physician. For example,
the physician may begin treatment using manufacturer-recommended
doses for the anti-VEGF agent, anti-inflammatory steroid and/or
NSAID, and make adjustments based on the physician's observations
of the effect of treatment. Further guidance is provided herein and
in the Examples. In addition, clinical trials can be conducted to
determine the doses that are effective to produce statistically
significant treatment effects when a population of patients is
treated.
[0022] As used herein "combination therapy" means the treatment of
a subject in need of treatment with a certain composition or drug
in which the subject is treated or given one or more other
compositions or drugs for the disease in conjunction with the first
and/or in conjunction with one or more other therapies, such as,
e.g., surgery. Such combination therapy can be sequential therapy
wherein the patient is treated first with one treatment modality
(e.g., drug or therapy), and then the other (e.g., drug or
therapy), and so on, or all drugs and/or therapies can be
administered simultaneously. In either case, these drugs and/or
therapies are said to be "co-administered." It is to be understood
that "co-administered" does not necessarily mean that the drugs
and/or therapies are administered in a combined form (i.e., they
may be administered separately or together to the same or different
sites at the same or different times).
[0023] The phrase "pharmaceutically acceptable" is employed herein
to refer to those compounds, materials, compositions and/or dosage
forms which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of human beings and
animals without excessive toxicity, irritation, allergic response,
or other problem or complication, commensurate with a reasonable
benefit/risk ratio.
[0024] As used herein, "treating" or "treatment" of a state,
disorder or condition (e.g., pterygium or keloid recurrence)
includes: (1) preventing or delaying the appearance of clinical or
sub-clinical symptoms of the state, disorder or condition
developing in a mammal that may be afflicted with or predisposed to
the state, disorder or condition but does not yet experience or
display clinical or subclinical symptoms of the state, disorder or
condition; and/or (2) inhibiting the state, disorder or condition,
including arresting, reducing or delaying the development of the
disease or a relapse thereof (in case of maintenance treatment) or
at least one clinical or sub-clinical symptom thereof; and/or (3)
relieving the disease, i.e., causing regression of the state,
disorder or condition or at least one of its clinical or
sub-clinical symptoms; and/or (4) causing a decrease in the
severity of one or more symptoms of the disease. The benefit to a
subject to be treated is either statistically significant or at
least perceptible to the patient or to the physician.
[0025] Treating can include inhibiting pterygium recurrence in a
subject. When pterygium recurrence is inhibited, pterygium is not
detectable for at least a predetermined time frame in the affected
eye of the subject where primary pterygium has been removed. For
example, a predetermined time frame may be, e.g., at least 1 month,
at least 2 months, at least 3 months, at least 4 months, at least 5
months, at least 6 months, at least 7 months, at least 8 months, at
least 9 months, at least 10 months, at least 11 months, or at least
12 months. Preferably, the predetermined time frame is at least 6
months.
[0026] Treating can include inhibiting keloid recurrence in a
subject. When keloid recurrence is inhibited, keloid is not
detectable for at least a predetermined time frame in the site
where primary pterygium has been removed. For example, a
predetermined time frame may be, e.g., at least 1 month, at least 2
months, at least 3 months, at least 4 months, at least 5 months, at
least 6 months, at least 7 months, at least 8 months, at least 9
months, at least 10 months, at least 11 months, or at least 12
months. Preferably, the predetermined time frame is at least 6
months.
Anti-VEGF Agents
[0027] The human VEGF-A gene is organized in eight exons.
Alternative exon splicing results in the generation of four main
VEGF isoforms, having, respectively, 121, 165, 189, and 206 amino
acids following signal sequence cleavage (VEGF121, VEGF165,
VEGF189, and VEGF206). VEGF165 is believed to be the most
physiologically relevant isoform. For a review, see Ferrara et al.,
Biochem. Biophys. Res. Commun., 2005, 333, 328-335. The amino acid
sequences of VEGF-A are well known in the art, and due to splice
variation, the sequences are numerous. By way of non-limiting
example, the following are exemplary and non-limiting GenBank.RTM.
Accession Nos. for human VEGF-A ("VEGF") amino acid sequences:
AAP86646.1, P15692.2, NP_001191313.1, NP_001165101.1,
NP_001165099.1, NP_001165097.1, NP_001165095.1, NP_001020539.2,
NP_003367.4, NP_001165093.1, NP_001020541.2, NP_001191314.1,
NP_001165100.1, NP_001165098.1, NP_001165096.1, NP_001165094.1,
NP_001028928.1, NP_001020540.2, NP_001020538.2, and
NP_001020537.2.
[0028] There are two VEGF receptor (VEGFR) tyrosine kinases (RTKs),
Flt-1, known also as VEGFR-1 and KDR, Flk-1, or VEGFR-2. There is
now agreement that VEGFR-2 is the major mediator of the mitogenic,
angiogenic, and permeability-enhancing effects of VEGF. For a
detailed review of the biological and signaling properties of the
VEGFR, see Ferrara, Endocr. Rev. 2004, 25, 581-611. The amino acid
sequences for the VEGFR are known in the art. By way of
non-limiting example, GenBank.RTM. accession numbers for VEGFR-1
amino acid sequences include (but are not limited to):
NP_001153503.1, NP_002010.2, NP_001153502.1, and NP_001153392.1.
The amino acid sequences for the VEGFR-2 are known in the art. By
way of non-limiting example, GenBank.RTM. accession numbers for
VEGFR-2 amino acid sequences include (but are not limited to):
NP_002244.1, AAC16450.1, and NP_001153503.1.
[0029] Disclosed herein are methods for treating pterygium or
keloid recurrence. The methods include administering an anti-VEGF
agent to a subject.
[0030] The present Examples demonstrate that the anti-VEGF antibody
bevacizumab can be used in the present methods. The antibody
bevacizumab and its VEGF-binding activity are reviewed in detail in
Ferrara et al., Biochem. Biophys. Res. Commun., 2005, 333, 328-335.
Bevacizumab may be administered to the eye in an amount of about
2.5 mg. However suitable ranges include, e.g., from about 0.5 mg to
about 10 mg, about 1 mg to about 5 mg, about 2 mg to about 4 mg,
and about 2 mg to about 3 mg. For example, the antibody can be
administered at a dosage in the range of about 0.5 mg/0.1 mL to 10
mg/0.1 mL in a total volume of 0.1 mL. Bevacizumab may be
administered to skin (e.g., for the inhibition of keloid
recurrence) at a dose about 5-15 mg. In one embodiment, the dose
for administration to site of keloid removal (e.g., for inhibition
of keloid recurrence) is about 10 mg).
[0031] It is to be appreciated, however, that the treatment method
described herein can also be performed using other anti-VEGF agents
(e.g., VEGF or VEGFR inhibitors, such as, but not limited to, other
anti-VEGF antibodies, drugs, prodrugs, small molecules, peptides,
nucleic acid inhibitors (e.g., siRNA, shRNA, antisense
oligonucleotides), fusion proteins, etc.), either that are known in
the art or that will be discovered or engineered in the future, so
long as the anti-VEGF agent has the ability to inhibit the action
of VEGF (e.g., human VEGF) and/or a VEGFR (e.g., VEGFR-1 and/or
VEGFR-2) (e.g., human VEGFR-1 or human VEGFR-2) (i.e., to inhibit
VEGF signaling). Assays for determining whether an antibody or
other agent interferes with VEGF signaling (either by inhibiting
VEGF or a VEGFR or the interaction between VEGF and its receptor),
for example, are well known in the art, and can be used to
determine whether an anti-VEGF agent interferes with VEGF signaling
and is therefore encompassed by the presently disclosed methods.
Non-limiting examples of such assays include the VEGF inhibition
assays described in Vicari et al., J. Biol. Chem., 2011, 286(15),
13612-25 and Brekken et al. Cancer Res., 2000, 60, 5117-24.
[0032] By way of non-limiting example, other anti-VEGF antibodies
and inhibitors that are known in the art, and, that can be used in
the methods disclosed herein include but are not limited to:
ranibizumab, pegaptanib, imatinib, vandetanib, sorafenib,
pazopanib, valatanib, vevasiranib, aflibercept, etanercept,
anecortave acetate (angiostatic steroid), VEGF-trap (a fusion
protein), squalamine lactate, erlotinib, gefitinib (small
molecules), Combretastatin A4 Prodrug (an
antitubulin/antiangiogenic agent), AdPEDF (Adenovector pigment
epithelium-derived factor), Candy (siRNA), protein tyrosine kinase
7 inhibitors (PTK7), lipolytic agents, TG100801, AG013958, AL39324,
AGN211745 (VEGF receptor blockers), anti-angiogenic VEGF-A(xxx)b
family, VEGF Trap (receptor decoy), protein kinase antibodies to
tyrosine kinase inhibitor receptors SIM010603, kinase domain
receptor antibodies (KDR1.3 and KDR2.6), GS101 aganirsen (an
antisense oligonucleotide against insulin receptor substrate aka
IRS-1), picropodophyllin (PPP), tetrameric tripeptide, tissue
kallikrein, KH906 (a recombinant human VEGF receptor protein
fusion), beta-adreno receptor blocker .beta.3-AR, nicotinic
acetycholine receptor antagonists, linomide analogue (Lin05),
morpholino oligomers (VEGFR1_MOe13), decursin, prorenin, vasohibin
and sirolimus. It will be appreciated that because the amino acids
sequences (as well as nucleic acid sequences encoding the amino
acid sequences) of VEGF and VEGFRs are known in the art, the
skilled artisan can readily design additional anti-VEGF agents for
use in the presently disclosed methods.
[0033] Dosage ranges for anti-VEGF agents, e.g., those disclosed
above, can be readily determined by the ordinarily skilled artisan,
and can, e.g., first be determined in animal models for determining
dosage, safety and efficacy according to standard methods known in
the art.
Anti-Inflammatory Steroids
[0034] Anti-inflammatory steroids are steroidal compounds that have
anti-inflammatory activity and include corticosteroids, including
glucocorticoids. Glucocorticoids bind to glucocorticoid receptors
in the cytoplasm which may increase the transcription of genes
coding for anti-inflammatory proteins, including lipocortin-1,
interleukin-10, interleukin-1 receptor antagonist and neutral
endopeptidase. Glucocorticoids also inhibit the expression of
multiple inflammatory genes, including genes for various cytokines,
enzymes, receptors and adhesion molecules. Barnes et al., Clin.
Sci., 1998, 94, 557-572.
[0035] Suitable anti-inflammatory steroids for injection into the
eye and suitable for use in the methods described herein include,
e.g., triamcinolone acetonide (Kenalog.RTM., e.g., Kenalog.RTM.-40,
available from Ranbaxy Laboratories Inc. (0.0147% spray),
Triesence.RTM. available from Alcon Laboratories Inc.,
Trivaris.RTM. available from Allergan Inc., Trianex.RTM. available
from Upsher Smith Laboratories Inc.). Suitable dosages of
triamcinolone acetonide range from 10 mg/mL to 40 mg/mL with volume
given range from 0.5 mL to 1 mL. These steroids can be co-injected
with other agents described herein (e.g., with the anti-VEGF
antibody). Further injectable anti-inflammatory steroids suitable
for use in the methods described herein are listed in Table 1.
TABLE-US-00001 TABLE 1 Commercially Available Injectable Steroids
Steroid Tradenames Methylprednisolone Depo-Medrol .RTM.;
Solu-Medrol .RTM. Dexamethasone Decadron .RTM.-LA; Decadron .RTM.
Phosphate Betamethasone Celestone .RTM. Hydrocortisone Various
Triamcinolone Aristocort .RTM., Kenalog .RTM.
[0036] Suitable steroids for topical administration to the eye
(e.g., as eye drops or ointment) include, e.g., prednisolone
acetate 1% (e.g., Pred Forte.RTM., Allergan Inc.) or difluprednate
0.05%. Suitable dosages for administration to humans include, e.g.,
one drop to eye with frequency ranging from every hour to every 24
hours depending on the degree of inflammation characterized by
redness, swelling, pain and intraocular pressures. Further
commercially available topical ocular steroids suitable for use in
the methods described herein are listed in Table 2.
TABLE-US-00002 TABLE 2 Commercially Available Topical Ocular
Steroids Concentration/ Steroid Tradenames Formulation Prednisolone
acetate Pred Forte .RTM. 1.0% suspension Econopred .RTM. 1.0%
suspension Plus (Alcon) 1.0% suspension AK-Tate .TM. 0.125%
suspension Pred Mild .RTM. 0.125% suspension Econopred .RTM.
Prednisolone Inflamase .RTM. 1.0% solution sodium Forte 0.5%
solution phosphate Metreton .TM. 0.125% solution Inflamase .RTM.
0.125% solution Mild AK-Pred .TM. Dexamethasone Maxidex .RTM. 0.1%
suspension alcohol Tobradex .RTM. ointment 0.1% suspension ointment
Fluorometholone Flarex .RTM. 0.1% suspension acetate Eflone .TM.
0.1% suspension Fluorometholone FML .RTM. 0.1% suspension alcohol
Fluor-Op .TM. 0.1% ointment FML .RTM.-Mild 0.25% suspension
Rimexolone Vexol .RTM. 1% suspension Medrysone HMS .RTM. 1.0%
suspension alcohol Lotoprednol Lotemax .RTM. 0.5% suspension
etabonate Alrex .RTM. 0.2% suspension Zylet .RTM. 0.5%
suspension
[0037] Suitable steroids for application to the skin include, e.g.,
alclometasone 0.05% cream (generic or Aclovate.RTM. 0.05% cream
available from PharmaDerm Inc.), diflorasone 0.005% cream,
prednicarbate 0.1% (generic, or Dermatop 0.1% cream or ointment
available from Sanofi-Aventis US LLC), and fluocinonide cream 0.1%
(e.g., Vanos, available from Medicis Inc.). Suitable dosages for
administration to humans include, e.g., 0.05% to 0.1%. Further
commercially available topical ocular steroids suitable for use in
the methods described herein are listed in Table 3.
TABLE-US-00003 TABLE 3 Commercially Available Topical Steroids
Potency Drug Super-High Betamethasone dipropionate, augmented
Clobetasol propionate Fluocinonide Flurandrenolide Halobetasol
propionate High Amcinonide Betamethasone dipropionate
Desoximetasone Diflorasone diacetate Fluocinonide Halocinonide
Triamcinolone Medium-High Amcinonide Betamethasone dipropionate
Betamethasone valerate Desoximetasone Diflorasone diacetate
Fluocinonide Fluticasone propionate Mometasone furoate
Triamcinolone acetonide Medium Clocortolone pivalate Fluocinolone
acetonide Hydrocortisone valerate Mometasone furoate Triamcinolone
acetonide Medium-Low Betamethasone valerate Desonide Fluocinolone
acetonide Fluticasone proprionate Hydrocortisone butyrate
Hydrocortisone probutate Hydrocortisone valerate Prednicarbate
Triamcinolone acetonide Low Aclometasone dipropionate Betamethasone
valerate Desonide Fluocinolone acetonide Triamcinolone acetonide
Very Low Hydrocortisone (base) Hydrocortisone acetate/Aloe vera
Hydrocortisone acetate/urea
Non-Steroidal Anti-Inflammatory Drugs (NSAIDs)
[0038] Non-steroidal anti-inflammatory drugs (NSAIDs) are
non-steroidal compounds that reduce inflammation. Most NSAIDs act
as nonselective inhibitors of the enzyme cyclooxygenase (COX),
inhibiting both the cyclooxygenase-1 (COX-1) and cyclooxygenase-2
(COX-2) isoenzymes. COX catalyzes the formation of prostaglandins.
Since COX-1 inhibition is believed to be associated with
gastrointestinal side-effects of NSAIDs, compounds that are
selective COX-2 inhibitors have also been developed.
[0039] Non-limiting examples of NSAIDs that can be administered to
the eye include, e.g., diclofenac 0.1%, ketorolac tromethamine
ophthalmic solution, 0.4% (Allergan Inc.) bromfenac ophthalmic
solution, 0.09% (Bausch and Lomb Inc.), nepafenac ophthalmic
suspension 0.1% (Alcon Laboratories Inc.) fluriprofen sodium 0.03%
solution and ketorolac tromethamine 0.5% solution. Suitable dosage
regimens for administration to humans include, e.g., 0.1% applied
one drop once a day, up to 4 times a day (every 6 hours).
[0040] Non-limiting examples of NSAIDs that can be administered to
the skin include, e.g., diclofenac 1% (topical) (e.g., Voltaren gel
1% from Novartis Pharmaceuticals Corp. or Pennsaid 1.5% solution
(topical) from Mallinckrodt Pharmaceuticals). Suitable dosages for
administration to humans include, e.g., 1% (topical) apply to skin
once a day to 40 mg (administer by injecting 1 mL into skin).
Methods of Treating Pterygium and Keloid Recurrence
[0041] Provided herein are methods for treating (e.g., inhibiting)
pterygium recurrence in an affected eye of a subject, the method
including (e.g., comprising, consisting essentially of, consisting
of) administering to the affected eye of a subject in need of such
treatment (1) at least one anti-VEGF agent (e.g., ranibizumab,
pegaptanib, anecortave acetate (angiostatic steroid), VEGF-trap (a
fusion protein), squalamine lactate, erlotinib, gefitinib (small
molecules), combretastatin A4 prodrug (an
antitubulin/antiangiogenic agent), AdPEDF (Adenovector pigment
epithelium-derived factor), Candy (siRNA), and TG100801 (VEGF
receptor blocker); and one or both of (2) an anti-inflammatory
steroid (e.g., triamcinolone acetonide, prednisolone acetate 1%, or
difluprednate 0.05%.) and (3) a non-steroidal anti-inflammatory
drug (NSAID) (e.g., diclofenac 0.1%, ketorolac tromethamine
ophthalmic solution 0.4%, bromfenac ophthalmic solution 0.09%, and
nepafenac ophthalmic suspension 0.1%).
[0042] In a specific embodiment, a method for treating pterygium
recurrence in an affected eye of a subject includes (e.g.,
comprises, consists essentially of, consists of) administering to
the affected eye of a subject in need of such treatment (1)
bevacizumab and (2) triamcinolone acetonide (or prednisolone
acetate 1%, or difluprednate 0.05%.) and/or (3) diclofenac 0.1% (or
ketorolac tromethamine ophthalmic solution 0.4%, bromfenac
ophthalmic solution 0.09%, or nepafenac ophthalmic suspension
0.1%).
[0043] In another specific embodiment, a method for treating
pterygium recurrence in an affected eye of a subject includes
(e.g., comprises, consists essentially of, consists of)
administering to the affected eye of a subject in need of such
treatment (1) bevacizumab; and (2) triamcinolone acetonide (or
prednisolone acetate 1%, or difluprednate 0.05%).
[0044] In another specific embodiment, a method for treating
pterygium recurrence in an affected eye of a subject includes
(e.g., comprises, consists essentially of, consists of)
administering to the affected eye of a subject in need of such
treatment (1) bevacizumab; and (2) diclofenac 0.1% (or ketorolac
tromethamine ophthalmic solution 0.4%, bromfenac ophthalmic
solution 0.09%, or nepafenac ophthalmic suspension 0.1%).
[0045] In yet another specific embodiment, a method for treating
pterygium recurrence in an affected eye of a subject includes
(e.g., comprises, consists essentially of, consists of)
administering to the affected eye of a subject in need of such
treatment (1) bevacizumab; (2) triamcinolone acetonide (or
prednisolone acetate 1%, or difluprednate 0.05%.); and (3)
diclofenac 0.1% (or ketorolac tromethamine ophthalmic solution
0.4%, bromfenac ophthalmic solution 0.09%, or nepafenac ophthalmic
suspension 0.1%).
[0046] Also provided herein are methods for treating (e.g.,
inhibiting) keloid recurrence, wherein the method includes (e.g.,
comprises, consists essentially of, or consists of) administering
to a site where a keloid has been removed in a subject (1) an
anti-VEGF agent (e.g., Ranibizumab, pegaptanib, Anecortave acetate
(angiostatic steroid), VEGF-trap (a fusion protein), squalamine
lactate, erlotinib, gefitinib (small molecules), combretastatin A4
prodrug (an antitubulin/antiangiogenic agent), AdPEDF (Adenovector
pigment epithelium-derived factor), Candy (siRNA), and TG100801
(VEGF receptor blocker). In certain aspects, the method further
includes administering to the site one or both of (2) an
anti-inflammatory steroid (e.g., alclometasone 0.005% cream,
diflorasone 0.005% cream, fluocinonide cream 0.1%, or prednicarbate
topical 0.1% cream or ointment); and (3) an NSAID (e.g., diclofenac
1% solution).
[0047] In a specific embodiment, a method for treating keloid
recurrence is provided, wherein the method includes administering
(1) bevacizumab. In certain embodiments, the therapy further
includes administering to the site (2) alclometasone 0.005% cream,
diflorasone 0.005% cream, fluocinonide cream 0.1%, or prednicarbate
topical 0.1% cream or ointment. In another embodiment, a method for
treating keloid recurrence is provided, wherein the method includes
administering to the site (of previous keloid removal) (1)
bevacizumab and (2) an NSAID diclofenac solution or gel (e.g., 1%
solution or gel). In another embodiment, a method for treating
keloid recurrence is provided, wherein the method includes
administering to the site (of previous keloid removal) (1)
bevacizumab, (2) alclometasone or diflorasone cream (e.g., 0.005%
cream), or prednicarbate topical 0.1% cream or ointment; and (3) an
NSAID (e.g., diclofenac 1% solution).
[0048] When a combination therapy is administered to a subject, it
will be appreciated that the agents administered as part of the
combination therapy can be administered in any order, e.g., as
determined appropriate, e.g., by the subject's physician. For
example, a subject may be first administered an anti-inflammatory
steroid (e.g., eye drop or skin cream), followed by injection or
other suitable route of administration of an anti-VEGF agent (e.g.,
as disclosed herein), followed by topical administration of an
NSAID (e.g., eye drop, ointment, or skin cream). However, in other
embodiments, the subject may be first administered an NSAID,
followed by injection or other suitable route of administration of
an anti-VEGF agent (e.g., as disclosed herein) alone, followed by
topical administration of an anti-inflammatory steroid, or followed
by injection (e.g., co-injection) or other suitable route(s) of
administration of the anti-VEGF agent and an injectable steroid. In
another embodiment, the subject can be first administered an
anti-VEGF agent, by a suitable route of administration, followed by
an NSAID and then an anti-inflammatory steroid, or the subject can
be first administered an anti-VEGF agent by a suitable route of
administration, followed by an anti-inflammatory steroid, followed
by an NSAID. In other embodiments, all two or all three agents
(anti-VEGF agent, anti-inflammatory steroid and/or NSAID) can be
administered together, e.g., at or near the same time, e.g., within
1, 2, 3, 4, 6, 8, 10, 12 or 24 hours of each other. It is also
contemplated that more than one anti-VEGF agent can be administered
in combination in the methods disclosed herein (either at the same
or different times, by the same or different route(s) of
administration).
[0049] The length of time between removal of the pterygium or
keloid and the first treatment can vary according to each patient's
needs. For pterygia, it is typical to wait at least a day or two
following the surgery, for example, to inject anti-VEGF antibody,
simply to give the patient time to recover. However, it is also
possible to inject the anti-VEGF antibody and/or injectable
steroid, as well apply topical steroid and/or NSAID on the day of
surgery. Treatment, e.g., with a combination therapy disclosed
herein can also begin, about 2 or more, about 3 or more, about 4 or
more, about 5 or more, about 6 or more, or about 7 or more days
after surgery, about 1 or 2 weeks or more, about 3 weeks or more,
about 4 weeks or more, or about 5 weeks or more after the surgery.
The treatment can also be commenced in preparation for surgery,
e.g., by administering one or more of the drugs within about 1 week
prior to the surgery.
[0050] Time between administrations of the different components
(antibody, steroid, and/or NSAID) of the combination therapy can
also range widely depending on each patient's needs. When two or
more agents of the combination therapy are not administered on the
same day, they can be administered about 1 or more, about 2 or
more, about 3 or more, about 4 or more, about 5 or more, about 6 or
more, or about 7 or more days apart, about 1 or 2 weeks or more,
about 3 weeks or more, about 4 weeks or more, or about 5 weeks or
more apart.
[0051] The methods disclosed herein provide an improved treatment
of pterygium and keloid. As discussed above, currently available
treatments typically result in recurrence after only a short period
of time (e.g., less than about 6 months). The presently disclosed
methods provide long-term treatments (including complete
inhibition) of pterygium and keloid recurrence (e.g., greater than
about 6, 7, 8, 9, 10 or 11 months following removal of the
pterygium or keloid). Typically, although not necessarily,
treatment with the combination therapy disclosed herein can
continue until a patient is determined to have no new blood vessel
growth and no pterygium recurrence for about 6 months from the time
of the last treatment. At that time, the patient may continue to
receive NSAID drug (e.g., NSAID eye drops), which may be
administered for up to, e.g., about one year.
Administration
[0052] Compositions and formulations including an antibody, steroid
and/or NSAID ("agent") as described herein can be administered
topically or parenterally, or by any other suitable methods known
in the art. The term "parenteral" includes injection or deposition
or sustained release via vehicles or devices (e.g., intravenous,
subconjunctival, subtenon, episcleral, intrascleral, subscleral,
intraperitoneal, epidural, intrathecal, intramuscular,
intraluminal, intratracheal, epidermal, intradermal, sub dermal or
subcutaneous). Moreover, the different agents administered in the
combination therapy disclosed herein may be administered by
different routes. For example, an antibody or other anti-VEGF agent
disclosed herein may be injection into the eye or skin, or applied
topically; an anti-inflammatory steroid and/or or NSAID may be
administered systemically (e.g., by injection), orally, and/or
topically, e.g., to the eye or skin.
[0053] While it is possible to use an agent disclosed herein for
therapy as is, it may be preferable to administer the agent as a
pharmaceutical formulation, e.g., in admixture with a suitable
pharmaceutical excipient, diluent, or carrier selected with regard
to the intended route of administration and standard pharmaceutical
practice. Pharmaceutical formulations include at least one active
compound, in association with a pharmaceutically acceptable
excipient, diluent, and/or carrier.
[0054] Administration of a composition or formulation can be once a
day, twice a day, or more often. Frequency may be decreased during
a treatment maintenance phase of the treatment, e.g., once every
second or third day instead of every day or twice a day. The dose
and the administration frequency can be adjusted based on the
judgment of the treating physician, for example taking into account
the clinical signs, pathological signs and clinical and subclinical
symptoms of a disease of the conditions treated with the present
methods, as well as the patient's clinical history. For example,
higher doses or frequency of administration, or a longer duration
of treatment may be indicated when a patient is showing symptoms of
pterygium or keloid recurrence (e.g., blood vessel growth), or if
the patient has a history of previous pterygium or keloid
recurrence. Specific examples of dosage and frequency are provided
in the Examples.
[0055] It will be appreciated that the amount of an agent disclosed
herein required for use in treatment will vary with the route of
administration, the nature of the condition for which treatment is
required, and the age, body weight and condition of the patient,
and will be ultimately at the discretion of the attendant physician
or veterinarian. Compositions will typically contain an effective
amount of the active agent(s), alone or in combination. Preliminary
doses can be determined according to animal tests, and the scaling
of dosages for human administration can be performed according to
art-accepted practices.
[0056] Length of treatment, i.e., number of days, will be readily
determined by a physician treating the subject; however the number
of days of treatment may range from about 1 day to about 365 days.
As provided by the present methods, and discussed below, the
efficacy of treatment can be monitored during the course of
treatment to determine whether the treatment has been successful,
or whether additional (or modified) treatment is necessary.
Kits
[0057] In certain embodiments, the present disclosure provides kits
for treating pterygium or keloid recurrence in a subject. The kits
can include an anti-VEGF antibody (e.g., bevacizumab, Ranibizumab,
Pegaptanib) or anti-VEGF agent (e.g., Anecortave acetate
(angiostatic steroid), VEGF-trap (a fusion protein), Squalamine
lactate (small molecule), Combretastatin A4 Prodrug (an
antitubulin/antiangiogenic agent), AdPEDF (Adenovector pigment
epithelium-derived factor), Candy (siRNA), and TG100801 (VEGF
receptor blocker)), and one or more of an anti-inflammatory steroid
(suitable for application to eye or skin) (e.g., triamcinolone
acetonide, prednisolone acetate 1% and/or difluprednate 0.05% (for
eye), or alclometasone 0.005% cream, diflorasone 0.005% cream,
fluocinonide cream 0.1%, and/or prednicarbate topical 0.1% cream or
ointment (for skin), and an NSAID (e.g., diclofenac 1% gel,
diclofenac 1.5% solution (topical)). The skilled artisan will
appreciate that the dosages of the above anti-inflammatory steroids
and NSAIDs may be varied without departing from the nature of the
present disclosure, and thus other dosages are also encompassed by
the present disclosure. The skilled artisan will know which dosages
of the anti-inflammatory steroids and NSAIDs disclosed herein may
be safely and effectively administered to a subject according to
the standard of care and knowledge in the art, and can include the
anti-inflammatory steroids and NSAIDs described herein.
[0058] In a specific embodiment, a kit includes bevacizumab; and
one or more of prednisolone acetate 1% and difluprednate 0.05% and
triamcinolone acetonide. In another specific embodiment, a kit
includes bevacizumab and diclofenac 0.1% (and/or ketorolac
tromethamine ophthalmic solution 0.4%, bromfenac ophthalmic
solution 0.09%, and/or nepafenac ophthalmic suspension 0.1%). In
another embodiment, the kit includes bevacizumab; one or more of:
prednisolone acetate 1%, difluprednate 0.05%, and triamcinolone
acetonide; and diclofenac 0.1%. In certain embodiments, the kit is
for treating pterygium. The kit can further optionally include
instructions for use. The kit can further optionally include (e.g.,
comprise, consist essentially of, consist of) syringes or
applicators preloaded with the above mentioned agents and/or vials
containing one or more of the agents.
[0059] In another embodiment, the kit includes bevacizumab; and one
or more of alclometasone 0.005% cream, diflorasone 0.005% cream,
fluocinonide cream 0.1%, and prednicarbate topical 0.1% cream or
ointment. In another embodiment, the kit includes bevacizumab; and
diclofenac 1% solution. In another embodiment, the kit include
bevacizumab; one or more of alclometasone 0.005% cream, diflorasone
0.005% cream, fluocinonide cream 0.1%, and prednicarbate topical
0.1% cream or ointment; and diclofenac 1% solution. In certain
embodiments, the kit is for treating keloid recurrence. The kit can
further optionally include instructions for use. The kit can
further optionally include syringes or applicators preloaded with
the above mentioned agents and/or vials containing one or more of
the agents.
[0060] The kits, regardless of type, will generally include one or
more containers into which the biological agents (e.g., inhibitors)
are placed and, preferably, suitably aliquotted. The components of
the kits may be packaged either in aqueous media or in lyophilized
form.
[0061] In accordance with the present invention, there may be
employed conventional molecular biology, microbiology, recombinant
DNA, immunology, cell biology and other related techniques within
the skill of the art. See, e.g., Sambrook et al., (2001) Molecular
Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory
Press: Cold Spring Harbor, N.Y.; Sambrook et al., (1989) Molecular
Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory
Press: Cold Spring Harbor, N.Y.; Ausubel et al., eds. (2005)
Current Protocols in Molecular Biology. John Wiley and Sons, Inc.:
Hoboken, N.J.; Bonifacino et al., eds. (2005) Current Protocols in
Cell Biology. John Wiley and Sons, Inc.: Hoboken, N.J.; Coligan et
al., eds. (2005) Current Protocols in Immunology, John Wiley and
Sons, Inc.: Hoboken, N.J.; Coico et al., eds. (2005) Current
Protocols in Microbiology, John Wiley and Sons, Inc.: Hoboken,
N.J.; Coligan et al., eds. (2005) Current Protocols in Protein
Science, John Wiley and Sons, Inc.: Hoboken, N.J.; Enna et al.,
eds. (2005) Current Protocols in Pharmacology John Wiley and Sons,
Inc.: Hoboken, N.J.; Hames et al., eds. (1999) Protein Expression:
A Practical Approach. Oxford University Press: Oxford; Freshney
(2000) Culture of Animal Cells: A Manual of Basic Technique. 4th
ed. Wiley-Liss; among others. The Current Protocols listed above
are updated several times every year.
[0062] The following examples are meant to illustrate, not limit,
the invention.
EXAMPLES
Materials and Methods
Treatment Protocol
[0063] A topical anesthetic (proparacaine 0.5% eye drops or Alcaine
manufactured by Alcon Laboratories Inc.) was applied to the eye
following recent pterygiectomy (from .about.5 days to 2 months
following surgery). The patients were examined to identify vascular
growth on eye conjunctiva. A syringe containing bevacizumab
(Avastin.RTM.) (Genentech/Roche) at a mixture of 2.5 mg/0.1 mL was
injected into the conjunctiva, and the plunger was drawn back to
determine if blood was present in the syringe hub. If blood was
present (indicating presence of blood vessel), the syringe needle
was withdrawn. Bevacizumab was injected into areas adjacent to
vascular growth, where blood vessels were dilated and numerous. If
bleeding occurred (e.g., in the form of subconjunctival or
subcutaneous hematoma), tamponade with Q-tips.RTM. was performed
until bleeding stopped; then injection was resumed. In some cases,
patients were injected with a combination of bevacizumab and
Kenalog.RTM.) (KENALOG-40 (triamcinolone acetonide), suspension in
an amount between 0.1 mL to 1 mL (1 mg to 40 mg).
[0064] Either before or after bevacizumab or bevacizumab/Kenalog
injection (time between treatments varied), patients were treated
with one or both of anti-inflammatory steroid eye drops and NSAID
eye drops. Anti-inflammatory steroid drops (prednisolone acetate 1%
(Allergan)) were given for 6 weeks with frequency range from once a
day to 4 times a day (every 6 hours) to every hour (24 drops/day).
NSAID eye drops (Diclofenac 0.1% (Bausch and Lomb)) drops (1 mg/mL
to 100 mg/mL) were given multiple times, 4 times a day every 6
hours for 3 to 6 months.
[0065] Patients were followed up to monitor for appearance of new
blood vessel growth in the eye. The presence of new vessel growth
was an indication for further treatment (e.g., with injection
and/or eye drops (steroid and/or NSAID). In some cases, during the
first surgery to remove pterygium, the pterygium was only partially
removed (e.g., to prevent necrosis). Thus, in some patients,
additional surgeries were subsequently performed to remove
remaining quadrants of the pterygium.
Treatment of Chronic Pterygium with Bevacizumab with or without
Combination Therapy
[0066] Five patients (Patients 1-5) underwent surgery to remove
primary pterygium (pterygiectomy) from the cornea of one or both
eyes followed by conjunctival grafting were treated as described
below.
1. Patient 1--Right Eye
[0067] A 35 year old patient with pterygium of the right eye was
treated by surgical removal of the pterygium and conjunctival
grafting. The timeline of the patient's treatment is shown in FIG.
1. Following recurrence of the pterygium, a second surgical removal
of the pterygium and conjunctival grafting procedure was performed
208 days after performing the first procedure. Following a further
recurrence of the pterygium, a third surgical removal of the
pterygium and conjunctival grafting procedure was performed 371
days after performing the first procedure.
[0068] The following drug treatments were given on the number of
days indicated after the third surgical removal and conjunctival
grafting procedure:
[0069] Day 47: Bevacizumab injection.
[0070] Day 164: Bevacizumab injection.
[0071] Day 222: Bevacizumab+Kenalog.RTM. injection.
[0072] Day 315: Bevacizumab injection.
[0073] Day 372: NSAID eye drops treatment started.
[0074] New blood vessel growth was observed at the time of the
bevacizumab injections.
[0075] No pterygium regrowth was observed as of about 520 days
following performance of the third surgical removal and
conjunctival grafting procedure with the drug treatment as
described above.
2. Patient 2--Left Eye
[0076] A 58 year old patient presented with pterygium of the left
eye. The timeline of the patient's treatment is shown in FIG. 2.
The patient was treated by surgical removal of the pterygium and
corneal regrafting. Following recurrence of the pterygium, a second
surgical removal of the pterygium and conjunctival grafting
procedure was performed 315 days after performing the first
procedure.
[0077] The following drug treatments were given on the number of
days indicated after the second surgical removal and conjunctival
grafting procedure:
[0078] Day 17: Bevacizumab injection.
[0079] Following a further recurrence of the pterygium, a third
surgical removal of the pterygium and conjunctival grafting
procedure was performed 486 days after performing the first
procedure.
[0080] The following drug treatments were given on the number of
days indicated after the third surgical removal and conjunctival
grafting procedure:
[0081] Day 14: NSAID eye drops treatment started.
[0082] Day 15: Bevacizumab and Kenalog.RTM. injection.
[0083] Day 32: Bevacizumab injection.
[0084] Day 47: Bevacizumab and Kenalog.RTM. injection.
[0085] Day 231: Bevacizumab injection.
[0086] In addition, a fourth surgical procedure, superior growth
removal, was performed 199 days following the third pterygium
removal and surgical grafting procedure. It is noted that superior
growth is not pterygium regrowth. The superior growth was a portion
of pterygium that was not removed in previous surgeries. Only one
quadrant of the pterygium could be removed each time (in each
surgery), because removing too many quadrants at once could cause
necrosis of the eye.
[0087] No pterygium regrowth was observed as of about 240 days
following performance of the third surgical removal and
conjunctival grafting procedure with the drug treatment described
above.
3. Patient 3
[0088] A 52 year old patient with pterygium of both the left eyes
was treated as described below. A timeline of the patient's
treatment is shown in FIGS. 3A and 3B.
Left Eye
[0089] The timeline for the treatment of the patient's left eye is
shown in FIG. 3A. The patient's left eye was treated by surgical
removal together with grafting of an amniotic membrane manufactured
by TOP Ophthalmics on Day 0. The following drug treatments were
given on the number of days indicated after the surgery was
performed on the left eye:
[0090] Day 53: Bevacizumab and Kenalog.RTM. injection into left
eye.
[0091] No NSAID drop was used in the left eye.
[0092] Pterygium completely regrew in the patient's left eye.
Right Eye
[0093] The timeline for the treatment of the patient's right eye is
shown in FIG. 3B. The patient's left eye was treated by surgical
removal together with grafting of an amniotic membrane manufactured
by IOP Ophthalmics. This procedure was performed 189 days following
surgical removal and grafting procedure performed on the left eye.
The following drug treatments were given on the number of days
indicated after surgery was performed on the right eye:
[0094] Day 1 Steroid eye drops treatment started.
[0095] Day 8: Bevacizumab injection into right eye.
[0096] Day 9: NSAID eye drops treatment started.
[0097] Day 28: Bevacizumab injection into right eye.
[0098] No regrowth of pterygium into the patient's right eye was
observed as of about 130 days following performance of the surgical
removal and amniotic membrane grafting procedure on the right
eye.
4. Patient 4--Right Eye.
[0099] A 70 year old patient presented with pterygium of the right
eye. The timeline of the patient's treatment is shown in FIG. 4.
The patient was treated on Day 0 by surgical pterygiectomy and
conjunctival grafting. The following drug treatments were given on
the number of days indicated after surgery was performed on the
right eye:
[0100] Day 1: Steroid eye drops first used in right eye.
[0101] Day 34: Bevacizumab and Kenalog.RTM. injections into right
eye.
[0102] Day 47: NSAID eye drop first used in right eye.
[0103] Day 78: Bevacizumab injection into right eye.
[0104] Day 90: Bevacizumab injection into right eye.
[0105] On day 74 following the initial surgery, a further
pterygiectomy procedure was carried out on the patient's right eye
to remove pterygium that could not be removed during the first
surgery because of scarring.
[0106] No regrowth of pterygium into the patient's right eye was
observed as of about 250 days following performance of first
surgical procedure to remove pterygium from the patient's right
eye.
5. Patient 5--Right Eye
[0107] A 66 year old patient presented with pterygium of the right
eye. A timeline of the patient's treatment is shown in FIG. 5. The
patient was treated by surgical pterygiectomy and amniotic membrane
grafting (manufactured by Biotissue). The following drug treatments
were given on the number of days indicated after surgery was
performed on the patient's right eye:
[0108] Day 1: Steroid eye drops and NSAID eye drops treatment
started.
[0109] Day 34: Bevacizumab and Kenalog.RTM. injections into right
eye.
[0110] Day 6: Bevacizumab injection into right eye.
[0111] Day 20: Bevacizumab injection into right eye.
[0112] No regrowth of pterygium into the patient's right eye was
observed as of about 90 days following performance of surgical
procedure to remove pterygium from the patient's right eye.
[0113] The various treatment regimens (combination therapies) and
results for patients 1-5 are summarized in Table 4, below:
TABLE-US-00004 TABLE 4 Steroid (injected Topical Bevacizumab and/or
NSAID Pterygium Patient Treatment eye drops) (eye drops) Recurrence
1 Right eye Yes Yes Yes No 2 Left eye Yes Yes Yes No 3 Right Eye
Yes Yes Yes No Left Eye Yes Yes No Yes 4 Right Eye Yes Yes Yes No 5
Right Eye Yes Yes Yes No
[0114] The above results demonstrated that pterygium regrowth was
inhibited when the patient received a combination therapy including
administration of bevacizumab, an anti-inflammatory steroid
(injected or topical), and a topical NSAID, and that the
combination therapy was superior to treatment with bevacizumab
alone.
[0115] A number of embodiments of the invention have been
described. Nevertheless, it will be understood that various
modifications may be made without departing from the spirit and
scope of the invention. Accordingly, other embodiments are within
the scope of the following claims.
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