Methods of Treating Chronic Neurogenic Inflammation Using Glucagon Like Hormone Retargeted ...

104 Pages·2013·6.76 MB·English

US 20100303791A1
(19) United States
(12) Patent Application Publication (10) Pub. No.: US 2010/0303791 A1
FRANCIS et al. (43) Pub. Date: Dec. 2, 2010
(54) METHODS OF TREATING CHRONIC (22) Filed: May 17, 2010
NEUROGENIC INFLAMMATION USING
Related US. Application Data
GLUCAGON LIKE HORMONE
RETARGETED ENDOPEPIDASES (60) Provisional application No. 61/182,452, ?led on May
29, 2009.
(75) Inventors: JOSEPH FRANCIS, ALISO
Publication Classi?cation
VIEJO, CA (US); DEAN G.
STATHAKIS, IRVINE, CA (US) (51) Int. Cl.
A61K 38/45 (2006.01)
A61K 38/43 (2006.01)
Correspondence Address:
C12N 9/96 (2006.01)
ALLERGAN, INC.
2525 DUPONT DRIVE, T2-7H (52) US. Cl. ...................... .. 424/945; 424/94.1; 435/188
IRVINE, CA 92612-1599 (US) (57) ABSTRACT
The present speci?cation discloses TVEMPs, compositions
(73) Assignee: ALLERGAN, INC.
comprising such toxins and methods of treating chronic neu
rogenic in?ammation in a mammal using such TVEMPs and
(21) Appl. No.: 12/781,284 compositions.
Patent Application Publication Dec. 2, 2010 Sheet 1 0f 7 US 2010/0303791 A1
Neuratransmitter
Reieasa
Vesinle
F1IAG..
Patent Application Publication Dec. 2, 2010 Sheet 2 0f 7 US 2010/0303791 A1
v
£>>2m0m I E@m5Eo2m5
Patent Application Publication Dec. 2, 2010 Sheet 3 0f 7 US 2010/0303791 A1
FEG. 2.
B-trefoil Domain
I. a-fold | | [3-fold | | y-fold |
| Enzymatic CleSIaiI.tv.H.e.a .. .g...e..T. ..r..a a.n sloca...t..i...o..nB...4 ../..l.. 3.|_ -5-- ------------B __i riiding [58/5s9i ngle'chain
LSSJ Form
Di-chain
Loop .
Naturally-occurrlng
Protease
l1 Translocation | Binding |
Di-chain
(I)
w Form
Patent Application Publication Dec. 2, 2010 Sheet 4 0f 7 US 2010/0303791 A1
HG. 3A.
P
I
I Binding I Translocation Ill-I Enzymatic I
I I
88
P Protease
{Tami
II Translocationll Binding I
FIG. 38.
P
I
I Binding I Enzymatic III-I Translocation I
LSSJ
P Protease
iafml
II Enzymatic I Binding I
Patent Application Publication Dec. 2, 2010 Sheet 5 0f 7 US 2010/0303791 A1
P
l
| Enzymatic |-|-| Binding | Translocation |
PProtease
4 Binding | Translocation I
(7)
a)
FIG. 45.
P
l .
|__Translocation HI| Binding | Enzymatic |
I 58 '
PProtease
4 Binding | Enzymatic I
ch
0)
Translocation
Patent Application Publication Dec. 2, 2010 Sheet 6 0f 7 US 2010/0303791 A1
F5. 4.
P
_ 1
| Enzymatlc 1 Binding III-l Translocation |
L 38 |
PProtease
| Enzymatic | Binding |
(b
a)
FIG. 4D.
P
l
|—Translocation | Binding Ill-I Enzymatic |
I 88 |
PProtease
| Translocation I Binding |
(I)
(I)
Patent Application Publication Dec. 2, 2010 Sheet 7 0f 7 US 2010/0303791 A1
Fl. P
1
| Enzymatic III-I Translocation | Binding I
PProtease
-| Translocation | Binding I
ch
cn
FIG. 55.
P
1
|—Translocation |I||| Enzymatic I Binding I
' ss '
PProtease
4 Enzymatic | Binding I
w
u)
Translocation
US 2010/0303791Al Dec. 2, 2010
METHODS OF TREATING CHRONIC secret the neurotransmitter into the synaptic cleft. There is a
NEUROGENIC INFLAMMATION USING great desire by the pharmaceutical industry to expand the use
GLUCAGON LIKE HORMONE of Clostridial toxin therapies beyond its current myo-relaxant
RETARGETED ENDOPEPIDASES applications to treat other nerve-based ailments, such as, e. g.,
various kinds of chronic pain, neurogenic in?ammation and
CROSS REFERENCE
urogentital disorders, as Well as other disorders, such as, e. g.,
pancreatitis. One approach that is currently being exploited to
[0001] This patent application claims priority pursuant to
expand Clostridial toxin-based therapies involves modifying
35 U.S.C. §l 19(e) to US. Provisional Patent Application Ser.
No. 61/182,452 ?led May 29, 2009, Which is hereby incor a Clostridial toxin so that the modi?ed toxin has an altered
porated by reference in its entirety. cell targeting capability for a non-Clostridial toxin target cell.
This re-targeted capability is achieved by replacing a natu
[0002] The ability of Clostridial toxins, such as, e.g., Botu
linum neurotoxins (BoNTs), Botulinum neurotoxin serotype rally-occurring targeting domain of a Clostridial toxin With a
targeting domain shoWing a preferential binding activity for a
A (BoNT/A), Botulinum neurotoxin serotype B (BoNT/B),
Botulinum neurotoxin serotype Cl (BoNT/Cl), Botulinum non-Clostridial toxin receptor present in a non-Clostridial
neurotoxin serotype D (BoNT/D), Botulinum neurotoxin toxin target cell. Such modi?cations to a targeting domain
serotype E (BoNT/E), Botulinum neurotoxin serotype F result in a Clostridial toxin chimeric called a TargetedVesicu
lar Exocytosis Modulating Protein (TVEMP) that is able to
(BoNT/F), and Botulinum neurotoxin serotype G (BoNT/G),
selectively bind to a non-Clostridial toxin receptor (target
and Tetanus neurotoxin (TeNT), to inhibit neuronal transmis
sion are being exploited in a Wide variety of therapeutic and receptor) present on a non-Clostridial toxin target cell (re
targeted). A Clostridial toxin chimeric With a targeting activ
cosmetic applications, see e. g., William J. Lipham, COSMETIC
ity for a non-Clostridial toxin target cell can bind to a receptor
AND CLINICAL APPLICATIONS OF BOTULINUM ToxIN (Slack,
Inc., 2004). Clostridial toxins commercially available as present on the non-Clostridial toxin target cell, translocate
pharmaceutical compositions include, BoNT/A preparations, into the cytoplasm, and exert its proteolytic effect on the
SNARE complex of the non-Clostridial toxin target cell.
such as, e.g., BOTOX® (Allergan, Inc., Irvine, Calif.), DYS
PORT®/RELOXIN®, (Beaufour Ipsen, Porton DoWn, [0005] Neurogenic in?ammation encompasses a series of
England), NEURONOX® (Medy-Tox, Inc., Ochang-myeon, vascular and non-vascular in?ammatory responses mediated
South Korea) BTX-A (Lanzhou Institute Biological Prod by a complex biological process that ultimately results in the
ucts, China) and XEOMIN® (MerZ Pharmaceuticals, local release of in?ammatory mediators and sensitiZing com
GmbH., Frankfurt, Germany); and BoNT/B preparations, pounds from sensory neurons. Upon insult by a noxious
such as, e.g., MYOBLOCTM/NEUROBLOCTM (Elan Phar stimulus, such as, e.g., a pathogen, damage to cells, or an
maceuticals, San Francisco, Calif.). As an example, irritant, in?ammation mediating and sensitiZing molecules,
BOTOX® is currently approved in one or more countries for such as, e. g., histamine, prostaglandins, leukotrienes, seroto
the folloWing indications: achalasia, adult spasticity, anal ?s nin, neutral proteases, cytokines, bradykinin and nitric oxide,
sure, back pain, blepharospasm, bruxism, cervical dystonia, are released from in?ammation mediating cells, such as, e. g.,
essential tremor, glabellar lines or hyperkinetic facial lines, mast cells, immune cells, vascular endothelial cells, and vas
headache, hemifacial spasm, hyperactivity of bladder, hyper cular smooth muscle cells. See Jennelle Durnett Richardson
hidrosis, juvenile cerebral palsy, multiple sclerosis, myo and Michael R. Vasko, Cellular Mechanisms ofNeurogenic
clonic disorders, nasal labial lines, spasmodic dysphonia, In?ammation, 302(3) J. Pharrnacol. Exp. Ther. 839-845
strabismus and VII nerve disorder. (2002), Which is hereby incorporated by reference in its
[0003] Clostridial toxin therapies are successfully used for entirety. These in?ammation mediating and sensitiZing mol
many indications. Generally, administration of a Clostridial ecules act on sensory neurons to stimulate the release of
toxin treatment is Well tolerated. HoWever, toxin administra in?ammation inducing molecules such as, e.g., neuropep
tion in some applications can be challenging because of the tides like substance P (SP) and calcitonin gene-related pep
larger doses required to achieve a bene?cial effect. Larger tide (CGRP), prostaglandins, and amino acids like glutamate,
doses can increase the likelihood that the toxin may move from the peripheral nerve endings. Upon release, these
through the interstitial ?uids and the circulatory systems, in?ammation inducing molecules are responsible for elicit
such as, e.g., the cardiovascular system and the lymphatic ing an in?ammatory response, typically characteriZed by
system, of the body, resulting in the undesirable dispersal of edema (sWelling secondary to plasma extravasation), hyper
the toxin to areas not targeted for toxin treatment. Such dis sensitivity (secondary to alterations in the excitability of cer
persal can lead to undesirable side effects, such as, e.g., inhi tain sensory neurons), and an erythema (redness and Warmth
bition of neurotransmitter release in neurons not targeted for secondary to vasodilation) Which extends beyond the site of
treatment or paralysis of a muscle not targeted for treatment. stimulation (the ?are response). Id. Because the underlying
For example, a patient administered a therapeutically effec in?ammatory symptoms are triggered by the activation of
tive amount of a BoNT/ A treatment into the neck muscles for primary sensory neurons and the subsequent release of
torticollis may develop dysphagia because of dispersal of the in?ammation inducing molecules, the response is termed
toxin into the oropharynx. As another example, a patient neurogenic in?ammation.
administered a therapeutically effective amount of a BoNT/A [0006] Normally, neurogenic in?ammation serves as a pro
treatment into the bladder for overactive bladder may develop tective mechanism by an organism to remove noxious stimuli
dry mouth and/or dry eyes. Thus, there remains a need for as Well as initiate the healing process for injured tissue. This
improved Clostridial toxins that are effective at the site of acute neurogenic in?ammation forms the ?rst line of defense
treatment, but have negligible to minimal effects in areas not by maintaining tissue integrity and contributing to tissue
targeted for a toxin treatment. repair. In fact, in the absence of acute neurogenic in?amma
[0004] A Clostridial toxin treatment inhibits neurotrans tion, Wounds and infections Would never heal and progressive
mitter release by disrupting the exocytotic process used to destruction of the tissue Would compromise the survival of
US 2010/0303791Al Dec. 2, 2010
the organism. However, severe or prolonged noxious stimu administering to the mammal in need thereof a therapeuti
lation results in a chronic neurogenic in?ammatory response cally effective amount of the composition, Wherein the com
provoking injury rather than mediating repair. This chronic position comprises a TVEMP including a retargeted peptide
neurogenic in?ammation has been implicated in the patho binding domain, a Clostridial toxin translocation domain and
physiology of a Wide range of unrelated disorders Which a Clostridial toxin enzymatic domain and Wherein adminis
underly a Wide variety of human diseases. tration of the composition reduces the release of an in?am
[0007] Attempts to treat chronic neurogenic in?ammation mation inducing molecule, thereby treating the mammal. Still
have met With limited success. This is due, in part, to the fact aspects of the present invention provide a use of a composi
that the etiology of chronic neurogenic in?ammation is a
tion for treating chronic neurogenic in?ammation in a mam
complex response based in part on the various in?ammation
mal in need thereof, the use comprising the step of adminis
inducing molecules and the multitude of in?ammation medi
tering to the mammal in need thereof a therapeutically
ating and sensitizing molecules that appear to elicit in?am
effective amount of the composition, Wherein the composi
mation via redundant mechanism. See Richardson & Vasko,
tion comprises a TVEMP including a retargeted peptide bind
302(3) J. Pharmacol. Exp. Ther. 839-845 (2002). Therefore,
ing domain, a Clostridial toxin translocation domain and a
compounds and methods that can prevent the chronic release
Clostridial toxin enzymatic domain and Wherein administra
of in?ammation inducing molecules from sensory neurons
tion of the composition educes a symptom of the chronic
Would be highly desirable for the treatment of chronic neu
rogenic in?ammation. neurogenic in?ammation, thereby treating the mammal.
[0008] The present speci?cation discloses TVEMP compo
sitions and methods for treating an individual suffering from BRIEF DESCRIPTION OF THE DRAWINGS
chronic neurogenic in?ammation. This is accomplished by
administering a therapeutically effective amount of a compo [0014] FIG. 1 shoWs a schematic of the current paradigm of
sition comprising a TVEMP to an individual in need thereof. neurotransmitter release and Clostridial toxin intoxication in
The disclosed methods provide a safe, inexpensive, out
a central and peripheral neuron. FIG. 1A shoWs a schematic
patient-based treatment for the treatment of chronic neuro for the neurotransmitter release mechanism of a central and
genic in?ammation.
peripheral neuron. The release process can be described as
[0009] Thus, aspects of the present invention provide a comprising tWo steps: 1) vesicle docking, Where the vesicle
composition comprising a TVEMP comprising a retargeted bound SNARE protein of a vesicle containing neurotransmit
peptide binding domain, a Clostridial toxin translocation ter molecules associates With the membrane-bound SNARE
domain and a Clostridial toxin enzymatic domain. A compo proteins located at the plasma membrane; and 2) neurotrans
sition comprising a TVEMP can be a pharmaceutical compo mitter release, Where the vesicle fuses With the plasma mem
sition. Such a pharmaceutical composition can comprise, in brane and the neurotransmitter molecules are exocytosed.
addition to a TVEMP, a pharmaceutical carrier, a pharmaceu FIG. 1B shoWs a schematic of the intoxication mechanism for
tical component, or both. tetanus and botulinum toxin activity in a central and periph
[0010] Other aspects of the present invention provide a eral neuron. This intoxication process can be described as
method of treating neurogenic in?ammation in a mammal, comprising four steps: 1) receptor binding, Where a
the method comprising the step of administering to the mam Clostridial toxin binds to a Clostridial receptor system and
mal in need thereof a therapeutically effective amount of a initiates the intoxication process; 2) complex internalization,
composition including a TVEMP comprising a retargeted Where after toxin binding, a vesicle containing the toxin/
peptide binding domain, a Clostridial toxin translocation receptor system complex is endocytosed into the cell; 3) light
domain and a Clostridial toxin enzymatic domain, Wherein chain translocation, Where multiple events are thought to
administration of the composition reduces the release of an occur, including, e.g., changes in the internal pH of the
in?ammation inducing molecule, thereby reducing a symp vesicle, formation of a channel pore comprising the translo
tom associated With chronic neurogenic in?ammation. cation domain of the Clostridial toxin heavy chain, separation
[0011] Other aspects of the present invention provide a of the Clostridial toxin light chain from the heavy chain, and
method of treating neurogenic in?ammation in a mammal, release of the active light chain and 4) enzymatic target modi
the method comprising the step of administering to the mam ?cation, Where the activate light chain of Clostridial toxin
mal in need thereof a therapeutically effective amount of a proteolytically cleaves its target SNARE substrate, such as,
composition including a TVEMP comprising a retargeted e.g., SNAP-25, VAMP or Syntaxin, thereby preventing
peptide binding domain, a Clostridial toxin translocation vesicle docking and neurotransmitter release.
domain, a Clostridial toxin enzymatic domain, and an exog [0015] FIG. 2 shoWs the domain organization of naturally
enous protease cleavage site, Wherein administration of the occurring Clostridial toxins. The single-chain form depicts
composition reduces the release of an in?ammation inducing the amino to carboxyl linear organization comprising an
molecule, thereby reducing a symptom associated With enzymatic domain, a translocation domain, and a retargeted
chronic neurogenic in?ammation. peptide binding domain. The di-chain loop region located
[0012] Still other aspects of the present invention provide a betWeen the translocation and enzymatic domains is depicted
manufacturing of a medicament for treating urogenital-neu by the double SS bracket. This region comprises an endog
rological disorder in a mammal in need thereof, the medica enous di-chain loop protease cleavage site that upon pro
ment comprising a TVEMP including a retargeted peptide teolytic cleavage With a naturally-occurring protease, such as,
binding domain, a Clostridial toxin translocation domain and e.g., an endogenous Clostridial toxin protease or a naturally
a Clostridial toxin enzymatic domain. occurring protease produced in the environment, converts the
[0013] Still aspects of the present invention provide a use of single-chain form of the toxin into the di-chain form. Above
a composition for treating chronic neurogenic in?ammation the single-chain form, the HCC region of the Clostridial toxin
in a mammal in need thereof, the use comprising the step of binding domain is depicted. This region comprises the [3-tre