TREATMENTS FOR RESISTANT ACNE
RELATED APPLICATIONS
[0001} This application claims benefit priority of of Indian Patent Application
No. 269/DEL/2014, filed January 29, 2014 and No. 3247/DEL/2014, filed
November 10, 2014, the content of both applications is incorporated herein by
reference in their entirety.
FIELD OF THE INVENTION
[0002] The present disclosure relates generally to novel molecules,
compositions, and formulations for treatment of bacterial infections in general and
more specifically to bacterial infections with antibiotic-tolerant pathogens.
BACKGROUND OF THE INVENTION
[0003] Acne vulgaris is a skin condition that affects over 85% of all people.
Acne is a term for a medical condition of plugged pores typically occurring on the
face, neck, and upper torso. Following are four primary factors that are currently
known to contribute to the formation of acne vulgaris; (1) increased sebum output
resulting in oily, greasy skin; (2) increased bacterial activity, normally due to an
overabundance of Propionibacterium acnes bacteria; (3) plugging
(hypercornifi cation) of the follicle or pilosebaceous duct; and (4) and
inflammation. The plugged pores result in blackheads, whiteheads, pimples or
deeper lumps such as cysts or nodules. Severe cases of acne can result in
permanent scarring or disfiguring.
[0004] Though acne vulgaris is multifactorial, a commensal skin bacteria ( .
acnes) plays a major role in the formation of acne lesion. It is an infection of
pilosebacious glands, oil glands in the skin. In most cases sudden breakouts of
acne can be correlated with sudden increased production of sebum in the affected
individual. During adolescence androgen hormones play a crucial role. It leads to
overproduction of sebum by the pilosebaceous gland. The situation gets further
accentuated by irregular shedding of dead skin from lining of hair follicles. As the
dead skin cells clump together in the oily environment, they can form plugs which
block the pores of the hair follicles. A pore clogged by the shedding skin is
referred to as a comedo.
[0005] This creates a very conducive anaerobic condition for P. acnes bacteria
to grow. Hyperproliferation of P. acnes leads to destruction of follicular walls and
it sends a danger signal to the host immune system. P. acnes may trigger an innate
immune reaction both in very early (microcomedogenic) and in late
(inflammatory) acne lesions via the activation of Toll like receptors 2 (TLR2) on
inante immune cells. TLR activation ultimately triggers the expression various
cytokines (like IL-6, 1L-8, L- 2, IL-17 etc) and chemokines that stimulate
recruitment of other host immune cells [Jeremy et al, 2003; Thibout et al, 2014].
Acne lesions range in severity from blackheads, whiteheads and pimples to more
serious lesions such as deeper lumps, cysts and nodules.
[0006] Although various over-the-counter products are commercially
available to counteract acne condition, such as anti-acne agents for topical use,
including salicylic acid; sulfur; lactic acid; glycolic acid; pyruvic acid; urea;
resorcinol; N-acetylcysteine; retinoic acid; isotretinoin; tretinoin; adapalene;
tazoretene; antibacterial s such as clindamycin, tetracyclines, and erythromycin;
vitamins such as folic acid and nicotinamide; minerals such as zinc; benzoyl
peroxide; octopirox; triclosan; azeiaic acid; phenoxyethanol; phenoxypropanol;
and flavinoids, these agents tend to lack in potential to mitigate the acne condition
and may have negative side effects when devised in conventional topical
formulations. A key challenge that has limited the use of topical formulations is
the absence of formulations with the desired physicochemical properties and high
drug loading, which maintains a concentration significantly higher tha the MIC
at the site of application by facilitating the right degree of penetration over time
but with minimal systemic exposure. A formulation that addresses these unmet
needs can be a significant advance in the treatment of acne.
[0007] Furthermore, as articulated in [Taglietti et al, 2008], when it comes to
the delivery of a drug to a specific site, topical formulations that are efficacious
are probably among the most challenging products to develop. Once the product is
applied on the skin, a complex interaction occurs between the formulation, the
active compounds, and the skin itself. The penetration of the active compound(s)
into the skin follows Pick's first law of diffusion, which describes the transfer rate
of solutes as a function of the concentration of the various ingredients, the size of
the treatment surface area, and the permeability of the skin. However, the skin's
permeability can be influenced by many factors, such as the drying, moisturizing,
or occluding effects of the excipients in the formulation, which, in combination,
can modulate the release of the product at the treatment site. In acne, the site of
action is inside the pilosebaceous unit and, therefore, an efficacious anti-acne
formulation should facilitate the penetration of the active compound(s) into this
extremely lipophilic environment. An effective topical formulation therefore
needs to provide a stable chemical environment in a suitable dispensing container
in order to accommodate multiple compounds that may have different, if not
incompatible, physicochemical characteristics [Tagleitti et a , 2008]. Once
applied, a topical formulation must interact with the skin environment, which can
influence the rate of the release of the compound(s) in order to achieve adequate
skin absorption, and exert additional physical effects on the skin, such as drying,
occluding, or moisturizing [Tagleitti et a , 2008]. For example, even if an active
agent is very potent, and is effective via a systemic route, in the case of topical
administration can behave completely differently, i.e. if the desired concentration
is not reached in the pilosebaceous (or skin) unit, it will not serve as an effective
anti-acne therapy. Similarly, a molecule or drug can behave entirely differently if
formulated with different compositions, which we demonstrate later in an
example. Similarly, two molecule or active agents may behave entirely differently
in the same formulation or composition. Therefore, every new molecule that
needs to be formulated for topical skin application poses a novel and independent
challenge as it is impossible to predict which composition and ratio of active and
excipients will provide the desired efficacy benefit.
[0008] Furthermore, an emerging condition is the evolution of strains of P.
acnes, which do not respond to theantibiotic agents such as clindamycin,
tetracyclines and erythromycincurrently approved for the treatment of acne.While
the earlier dogmawas that antibiotics failure arises due to selection of 'resistant'
strains, i.e. a mutation resulting in alteration of the target of the
antibioticrendering it ineffective, emerging evidence suggests that antibiotic
failure is more complex than this simple understanding. The assumption was that
if resistance develops, i.e. the target of an antibiotic is altered, it is possible to
treat the condition by changing to an alternative antibiotic, the target of which is
still intact in the bacteria. However, recent knowledge has rendered this
assumption as false. For example, Regoes et al, 2004 demonstrates that even in
the absence of any resistance, a subset of bacteria can just exhibit tolerance to an
antibiotic, i.e. not undergo lysis. This could arise due to physiological (metabolic)
and morphological changes observed in bacteria exposed to antibiotics.For
example, in a study published in Science, [Miller et al, 2004] showed that a
transient induction of SOS response by ampicillin can protect E. coli against the
bactericidal effects of ampicillin Regoes et al, 2004 suggested that tolerance
mechanisms could cross over between some antibiotics, i.e. Antibiotic A might be
rendered ineffective due to development of resistance, but it is possible that
Antibiotic B, which has an entirely different target/mechanism of action, and is
shown to be active in a different or sensitive bacterial strain, may also be rendered
ineffective in the resistant strain due to shared tolerance mechanisms. Indeed,
massive changes in gene expression leading to alteration in the syntheses of
proteins of metabolic and stress response pathways and cell division during
exposure of E. coli to ampicillin and ofloxacin have been observed, and a number
of these alterations in the gene expression levels were shared between bacteria
exposed to ampicillin and ofloxacin, suggesting a bacteria not respondingto
ampicillin may not respond to ofloxacin although both agents have different
targets. We saw a similar observation in screening a library of antibiotics against
different strains of P. acnes that are sensitive or non-responsive to clindamycin (a
lincosamide). As shown in Fig. 1A, the strain of P. c on responsive to
clindamycin also showed increased survival capability in the presence of
roxithromycin (a nac iide) which targets a different site from clindamycin.
[Keren et al, 2004]. Specialized persister cells and the mechanism of multidrug
tolerance in Escherichia coli. J . Bacteriol. 186 8172-8 180) suggested that random
fluctuations in gene expression are responsible for the formation of specialized
persister cells. As argued by Regoes et al, 2004, phenotypic tolerance to antibiotic
could actua y prevent clearance. As a result, while there remains a need in the art
for compositions, formulations and methods for treating acnethat is not
responding to the currently used agents, especially clindamycin-, minocydine-,
erythromycin-, and/or doxycycline, the probability of tolerance makes
itimprobable to predict a drug that may work against P. acnes.
[0009] Furthermore, it is increasingly becoming evident that subtle changes in
chemical structure of a molecule can dramatically change activity of the
molecules against target protein. For example erythromycin (a macrolide) and
clindamycinbind to similar 50S ribosomal unit but crystal structure [Schulzen et
al, 200 showed different mode of binding between the agents and amino acid
residues present in SOS ribosomal sub-unit. There are known bacterial strains of
P. acnes that are resistant to clindamycin but can be either non-responsive or
susceptible to erythromycinand vice versa. Interestingly, telithromycin, which is a
semisynthetic derivative of erythromycin works well in a bacterial strain that is
resistant to both erythromycin and clindamycin [Beitru et al, 2003] Similarly, in
another example, the introduction of 8-chloro group dramatically enhanced the
potency of moxifioxacin but a similar change in gatifloxacin had no effect
agains S aureus, S. pneumonia, and E. coli. Additionally, molecules of the same
class can have different affinity for the same protein target but in different
bacteria. For example it has been found that both besifloxacin and moxifioxacin
effectively bind to DNA gyrase than ciprofloxacin in S. aureus. n contrary
ciprofloxacin binding towards DNA gyrase is more effective in E.coli than
moxifioxacin or besifloxacin. Similarly, besifloxacin is found to be best effective
molecule against S. pneumonia followed by moxifioxacin and ciprofloxacin.
[Cambau et al, 2009]. t is therefore not possible to predict the activity of a
moleculeagainst a bacteria or microbe based on its similarity in structure another
drug that shows activity against the same microbe or a different microbe, even
though they might have similar mechanisms of action. Indeed, as shown in Figure.
1, we observed that molecules that were verisimilar in structure had completely
distinct activity against P. acnes, i.e. where one was inactive the other was very
potent against both ciindamycin-susceptible and -non-resistant P. acnes (Fig. 1 A
and IB). In another example, which we discuss later, we observed a nonlincosamide
molecule that was very effective in a P. acnes strain resistant to
clindamycin but not active in a ciindamycin-sensitive P. acnes (Fig. 1A and IB).
The identification of an effective drug that works against both sensitive as well as
clindamycin-, minocycline-, erythromycin-, or doxyeycline-nonresponder P.
acnes therefore emerges through serendipity during systematic screening in P.
acnes.
[0010] Furthermore, while an emerging problem is the development of
resistant strains of microbes that are not responding to antimicrobial compounds
and compositions well known in the art, there remains a need in the art for a more
effective antibiotic that not only works against resistant microbes but also reduces
the risk of development of resistance by the microbes to this new antibiotic. Thus
molecules that are efficacious antibiotics and also 'prevent' or reduce the
development of resistance can be a major advancement in the treatment of
microbial diseases.
[00 ] The inflammatory character of acne has been correlated with the host
immune response targeting Propionihacterium acnes. In vitro studies demonstrate
that P. acnes whole cells or cell fractions stimulate cytokine and matrix
metalloproteinase release from immune cells, keratinocytes, and sebocytes [Kim
et al, 2002; Liu et al., 2005: Nagy et al., 2006; Lee et al., 2010] Though P. acnes
are ong been present in the follicular area , they come in direct contact with
immune cells in dermis on y after follicular rupture, The innate immune system
recognizes P. acnes via TLR2 [Kim et al., 2002], leading to the secretion of
inflammatory cytokines, including IL-6.IL-8 L-12 etc. Follicular rupture
happens very late in the disease process. But there are multiple evidences which
suggest that the adaptive immune response also has a significant role in the
inflammation observed even in early stages of acne, resulting from the recruitment
of activated T helper 1 (Thl) lymphocytes to early acne lesions [Mouser et al.,
2003].A potential treatment of acne therefore needs to resolve inflammation, and
should be able to target these inflammatory pathways.
[0012] An ideal treatment for acne therefore needmolecules that can work at
two or more targets. Molecules that work against both antibiotic-sensitive as well
as clindamycin-, minocycline-, erythromycin- and doxycycHne-tolerant or nonresponsivestrains
of P. acnesand ca additionally inhibit the P. c« , -activated
inflammatory mediator/s, or molecules that target two or more cellular targets in
these microbes while additionally exerting an inhibitory effect on the P. acnesactivated
inflammatory mediator/s, and is formulated in an optimal formulation
that enables the desired concentration of the active agent on the skin or
pilosebaceous region following topical application can emerge as a powerful
strategy for the treatment of acne.
SUMMARY
DART
[0013] A series of novel DART (Dual Action Rational Therapeutics)
molecules were designed and synthesized for treatment of bacterial infections
caused by both susceptible and resistant gram positive and gram negative bacteria
and specially for curing acne and different skin and skin structure infections and
additionally prevent the development of resistance. DART molecules can mount
its activity through two distinct mechanisms of action in a microbe (such as a
bacteria), and create less chance in mutation development at both target sites in
the bacteria. Additionally, it can also act at the host level by modulating the
immune response, such as altering the levels of inflammatory cytokines.
[0014] The design of DART comprises of two active domains. The two active
domains can be selected from different families, for example,P-lactam, b-lactam
derivatives, 2- and 4-quinolones, quinolones having halogenated atom specially
fluorine atom attached at C-6 or C-7 position of the central ring system,
fluoroquinolone with halogenated atom specially chlorine atom attached at C-8
position of the central ring system, tetracycline, oxazolidinone, hydroxypyridones,
derivatives of hydroxypyridones, pieuromutifin, azoles, nitroimidazoles,
monoxycarbolic acid class, fusidic acid, sulfonamide, sulfonamide derivatives,
retinoids, different fatty acids (saturated, unsaturated), propylene glycol and
glycerol derivatives of different fatty acids and a strategic combination from each
of the families. The design was made strategically by arranging the two active
domains in the right steric arrangement for both the active domains to maintain
their function against bacteria or fungus. Overall these molecules possess faster
bacterial killing with reduction in inflammation and activity against resistant
pathogens. These molecules also show a lower risk of development of resistance.
[0015] In some embodiments, the DART molecule has at least two chemical
domains. Each of said chemical domains binds to a distinct or different active site
in a target cell. In a preferred embodiment, a third chemical domain may be
present. In a further preferred embodiment, said two chemical domains may be
bound together through a said third domain. In some embodiments, the DART
molecule has at least two distinct or different anti-bacterial mechanisms of action.
In some embodiments, the DART molecule has at least two distinct or different
anti-acne mechanisms of action. Without limitations, the DARTs can act on the
same target or on different targets, for example, the bacteria and the host. In some
embodiments, the DART acts on at least two different targets. In some
embodiments, at least one of the targets is different than that affected by
conventional antibiotics.
[0016] n some embodiments, the DART molecule has a b-lactam ring and a
quinolone nucleus, or a quinolone nucleus and a nitro-heterocycle, or a b-lactam
ring and a nitroheterocycle
[0017] In some embodiments, the DART has at least two distinct antibacterial
mechanisms of action, for example inhibits DNA gyrase or
topoisomerase IV and transpeptidase-mediated cross-linking of peptidoglycans;
inhibits isoprenyl pyrophosphate and transpeptidase-mediated cross-linking of
peptidoglycans; inhibits isoprenyl pyrophosphate and DNA gyrase of
topoisomerase IV; inhibits folate synthesis and DNA gyrase of topoisomerase IV;
inhibits folate synthesis and transpeptidase-mediated cross-linking of
peptidoglycans; inhibits DNA gyrase or topoisomerase IV and the 30S ribosomal
sub-unit in bacteria; inhibits DNA gyrase or topoisomerase IV and the 50S subunit
in bacteria; inhibits transpeptidase-mediated cross-linking of peptidoglycans
and the 30S or the 50S ribosomal sub-unit in bacteria; inhibits folate synthesis and
the 30S or the 50S sub-unit in bacteria; or inhibits isoprenyl pyrophosphate and
the 3OS or the 50S sub-unit in bacteria, or causes DNA modification, such as
inducing DNA nickswhile inhibiting the induction of negative supercoils in
DNA;or altering the fluidity of the cell membrane while exerting an activity on
the DNA; or altering the levels of metal ions in a cell while inducing DNA
changes .in some embodiments, the first mechanism of action is an anti-bacterial
action and the second mechanism of action is anti-inflammatory or
immunomodulatory
[0018] In some embodiments, the DART molecule has at least two distinct
treating acne mechanisms of action and modulates at least two different targets.
In some embodiments, the first mechanism is an antibacterial action a d the
second mechanism of action is inhibition of keratinocyte proliferation and
differentiation n some embodiments, the DART molecule has two distinct acne
treating mechanisms of action and whereinthe first mechanism is an antibacterial
action and the second mechanism of action is anti-inflammatory. In some
embodiments, the DART molecule is effective against forms of Propionbacterium
acnes that respond poorly to clindamycin-, or doxycycline-, or erythromycin-, or
minoeyeline-eontaining anti-acne products . In some embodiments, they
areeffective against one or more of clindamycin-, minocycline-, erythromycin-,
and/or doxycycline- tolerant or resistant strains of Propionbacterium acnes. I
some embodiments, it prevents the development of resistance in P acnes,
[0019] In some embodiments, the DART molecule has at least two distinct
anti-bacterial mechanisms of action and modulates at least two different targets
against a pathogen. Non limiting examples of such pathogens are : Bartonella
henselae, Borrelia burgdorferi, Campylobacter jejuni, Campylobacterfetus,
Chlamydia trachomatis, Chlamydia pneumoniae, Chylamydia psittaci, Simkania
negevensis, Escherichia coli (e.g., 0157:H7 and 88), Ehrlichia chafeensis,
Clostridium botulinum, Clostridium perfringens, Clostridium tetani, Enterococcus
faecalis, Haemophilias influenzae, Haemophilius ducreyi, Coccidioides immitis,
Bordetella pertussis, Coxiella burnetii, Ureaplasma urealyiicum, Mycoplasma
geniialium, Trichomatis vaginalis, Helicobacter pylori, Helicobacter hepaiicus,
Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium hovis,
Mycobacterium africanum, Mycobacterium leprae, Mycobacterium asiaticum,
Mycobacterium avium, Mycobacterium celatum, Mycobacterium celonae,
Mycobacterium fortuitum, Mycobacterium genavense, Mycobacterium
haemophilum, Mycobacterium intracellular e, Mycobacterium kansasii,
Mycobacterium malmoense, Mycobacterium marinum, Mycobacterium
scrofulaceum, Mycobacterium simiae, Mycobacterium szulgai, Mycobacterium
ulcerans, Mycobacterium xenopi, Corynebacterium diptheriae, Rhodococcus equi,
Rickettsia aeschlimannii, Rickettsia africae, Rickettsia conorii, Arcanobacterium
haemolyticum, Bacillus anthracis, Bacillus cereus, hysteria monocytogenes,
Yersinia pestis, Yersinia enterocolitica, Shigella dysenteriae, Neisseria
meningitides, Neisseria gonorrhoeae, Streptococcus hovis, Streptococcus
hemolyticus, Streptococcus mutans, Streptococcus pyogenes, Streptococcus
pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus
pneumoniae, Staphylococcus saprophyticus, Vibrio cholerae, Vibrio
parahaemolyiicus, Salmonella typhi, Salmonella paratyphi, Salmonella
enteritidis, Treponema pallidum, Candida, Cryptcooccus, Cryptosporidium,
Giardia lamblia, Microsporidia, Plasmodium vivax, Pneumocystis carinii,
Toxoplasma gondii, Trichophyton mentagrophytes, Enterocytozoon bieneusi,
Cyclospora cayetanensis, Encephalitozoon hellem, Encephalitozoon cuniculi,
among other bacteria, archaea, protozoa, and fungi.
[0020] In some embodiments, ihe first and second domains independently
have antibacterial activity against a Staphylococcus species. Examples of
Staphylococcus species include, but are not limited to, S. aureus group (e.g., S.
aureus, S . simiae), S. auricularis group (e.g., S. auricularis), S. carnosus group
(e.g., S . carnosus, S. condimenti, S . massiliensis, S. piscifermentans, S. simulans),
5 . epidermidis group (e.g., S. capitis, S. caprae, S. epidermidis, S.
saccharolyticus), S. haemolyiicus group (e.g., S . devriesei, S. haemolyticus, S.
hominis), S . hyicus-intermedius group (e.g., S. chromogenes, S. felis, S delphini,
S. h.yicus, S. intermedius, S. lutrae, S. microti, S. muscae, S. pseudintermedius, S.
rostri, S. schleiferi), S. lugdunensis group (e.g., S. lugdunensis), S. saprophytics
group (e.g., S. arlettae, S. cohnii, S. equorum, S. gaUlnarum, S. kloosii, S. leei, S.
nepalensis, S. saprophytics, S. succinus, S. xylosus), S. sciuri group (e.g., S.
jleurettii, S . lentus, S. sciuri, S. stepanovicii, S. vitulinus), S. simulans group (e.g.,
S. simulans), and S. warneri group (e.g., S. pasteuri, S. warneri).
[0021] Without limitations, the DARTS can be in the form of particles,
powders, suspensions, dispersions, emulsions, liposomes, micelles, globules,
solutions, vesicles, aggregates, creams, gels, and the like
[0022] The disclosure also provides formulations comprising DARTs as the
active pharmaceutical ingredient (API).
Antibiotics
[0023] The disclosure also provides formulations comprising antibiotic
agents, which are not DARTs, as the API.In some embodiments, the antibiotic
agent is a8-chloro fluoroquinolone. Exemplary 8~ehloro fluoroquinolones
include, but are not limited to, besifloxacin, clinafloxacin and sitafloxacin. In
some embodiments, the formulation comprises besifloxacin as the API.
[0024] In various embodiments, the API can be micronized, suspended, or
solubilized. In some embodiments, the API can be in the form of particles,
powders, suspensions, dispersions, emulsions, liposomes, micelles, globules,
solutions, vesicles, aggregates, and the like. In some embodiments, the API ca in
the form of a drug carrier.
[0025] In some embodiments, the API can be coated. In some other
embodiments, the API can be uncoated.
[0026] Without limitations, the formulation can be in a form selected from the
group consisting of lotions, creams, gels, emulgel, oils, serums, powders, sprays,
ointments, solutions, suspensions, dispersions, pastes, foams, peels, films, masks,
patches, sticks, rollers, cleansing liquid washes, cleansing solid bars, pastes,
foams, powders, shaving creams, impregnated fabric ), and the like. In some
I
embodiments, the formulation is in a form selected from the group consisting of
gel, cream, spary, face wash, soap bar, body wash, lotion., suspended drug loaded
gel, suspended drug loaded cream, and any combinations thereof.
[0027] In some embodiments, the API or the formulation can be used to treat
acne not responding to antibiotics. Specifically, it exerts greater efficacy against
forms of Propionbacterium acnes that respond poorly to clindamycin-, or
doxycyc!ine-, or erythromycin-, or minocycline-containing anti-acne products.
[0028] In some embodiments, the API or the formulation can be used to treat
acne by exerting an anti-inflammatory effect.
[0029] In some embodiments, the API or the formulation can be used to treat
acne by killing strains of Propionbacterium acne that are sensitive toone or more
of clindamycin-, minocycline-, erythromycin-, and/or doxycyclineand additionally
exerting a greater efficacy by inhibiting P. «e.v-mediated inflammatory
pathways (i.e. dual mechanisms of action).
Combinations
[0030] The disclosure also provides formulations comprising a combination of
two or more antibiotic agents. For example, an 8-chloro fluoroquinolone in
combination with another anti-acne agent. In some embodiments, the formulation
comprises two or more different 8-chloro fluoroquinolones n some
embodiments, the formulation comprises besifloxacin and a retinoid, such as
adapalene.
[0031] In some embodiments, the formulation comprises an anti-acne agent
and an anti-inflarnmatory agent. For example, the formulation can comprise an 8-
chloro fluoroquinolone and an anti-inflammatory agent.
[0032] In some embodiments, the two or more antibiotic agents can be a
DART molecule or two or more different DART molecules. In some
embodiments, one of the two or more antibiotic agent is a DART and the other is
not a DART molecule.
[0033] As described herein, the disclosure provides formulations comprising
DART and/or non-DART antibitotic agent as the API. As such, exemplary API's
for the formulations include DARTs, anti-bacterial, anti-fungal and anti-acne
agents. In some embodiments, the API can be in the form of a drug carrier, i.e.,
the API can be nanotized, coated, made into vesicles, liposome, emulsions, and
the like for the formulation. Without limitations the formulation or the
composition can be formulated for administration by any appropriate route known
in the art including, but not limited to, topical (including buccal and sublingual)
and oral or parenteral routes, including intravenous, intramuscular, subcutaneous,
transdermal, airway (aerosol), pulmonary, and nasal administration.
[0034] The DARTs and formulations disclosed herein can be used for treating
bacteria! infections due to Gram-positive or Gram-negative bacteria.
Additionally, the DARTs are effective against resistant forms of pathogens.
Furthermore, the DARTs are effective in preventing the development of resistant
forms of pathogens. Thus, the DARTs and formulations disclosed herein can be
used for treating antibiotic tolerant or resistant bacterial infections. Exemplary
bacterial infections include, but are not limited to, infection by Bartonella
henselae, Borrelia burgdorferi, Campylobacter jejuni, Campylobacterjetus,
Chlamydia trachomatis, Chlamydia pneumoniae, Chylamydia psittaci, Simkania
negevensis, Escherichia coli {e.g., 0157:H7 and K88), Ehrlichia chafeensis,
Clostridium botulinum, Clostridium perfringens, Clostridium tetani, Enterococcus
faecalis, Haemophilius influenzae, Haemophilius ducreyi, Coccidioides immitis,
Bordetella pertussis, Coxiella burnetii, Ureaplasma urealyticum, Mycoplasma
genitalium, Trichomatis vaginalis, Helicobacter pylori, Helicobacter hepaticus,
Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium bovis,
Mycobacterium africanum, Mycobacterium leprae, Mycobacterium asiaticum,
Mycobacterium avium, Mycobacterium celatum, Mycobacterium celonae,
Mycobacterium fortuitum, Mycobacterium genavense, Mycobacterium
haemophilum, Mycobacterium intracellular e, Mycobacterium kansasii,
Mycobacterium malmoense, Mycobacterium marinum, Mycobacterium
scrofulaceum, Mycobacterium simiae, Mycobacterium szulgai, Mycobacterium
ulcerans, Mycobacterium xenopi, Corynebacterium diptheriae, Rhodococcus equi,
Rickettsia aeschlimannii, Rickettsia africae, Rickettsia conorii. Arcanobacterium
haemolyticum, Bacillus anthracis, Bacillus cereus, hysteria monocytogenes,
Yersinia pestis, Yersinia enterocoliiica, Shigella dysenteriae, Neisseria
meningitides, Neisseria gonorrhoeae, Streptococcus hovis, Streptococcus
hemolyticus, Streptococcus mutans, Streptococcus pyogenes, Streptococcus
pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus
pneumoniae, Staphylococcus saprophyticus, Vibrio cholerae, Vibrio
parahaemolyticus, Salmonella typhi, Salmonella paratyphi, Salmonella
enteritidis, Treponema pallidum, Candida, Cryptcooccus, Cryptosporidium,
Giardia lamhlia, Microsporidia, Plasmodium vivax, Pneumocystis carinii',
Toxoplasma gondii, Trichophyton mentagrophytes, Enter ocyiozoon bieneusi.
Cyclospora cayetanensi's, Encephalitozoon hellem, Encephalitozoon cimiculi,
among other bacteria, archaea, protozoa, and fungi. In some embodiments,
infection is with a Staphylococcus species.
[0035] n some embodiments, the DARTS and formulations disclosed herein
ca be used for treating acne. In some embodiments, the DARTS and
formulations disclosed herein are effective against forms of Propionbacterium
acnes that respond poorly to clindamycin-, or doxycycline-, or erythromycin-, or
rnmocyciine-eontammg anti-acne products. In some embodiments, the DARTs
and formulations disclosed herein are effective against one or more of
clindamycin-, minocycline-, erythromycin-, and/or doxycycline- tolerant or
resistant strains of Propionbacterium acnes. For treating infections, the DART or
the formulation disclosed herein can be administered once or daily to the subject
as a single dose or a plurality of doses.
BRIEF DESCRIPTION OF THE DRAWINGS
[0036] Figs 1A a d B shows dose response curves of different antibiotics
against both MTCC1951 and CCARM 9010 strains of P. acnes. While
MTCC1951 is killed by clindamycin, the drug has no effect on CCARM9010.
Different antibiotics behave differently and unpredictably on the different strains
belong to a different family from Clindamycin.
[0037] Figs, 1C and D are line graphs showing concentration efficacy curve
of DART compounds 90, 91, 94, 113, 115 a d 116 in both clindamycinsusceptible
(MTCC 1951) (Fig. 1C) and clindamycin-nonresponsive (CCARM
9010) (Fig ID) P. acnes strains. The molecules have different and unpredictable
activity against MTCC 1951 and CCARM9010 strains of P. acnes. Compound 9 1
showed highly efficacious bacterial killing profile for both bacterial strains.
Compound 90 shows activity in P. acnes that do not respond to clindamycin but is
ineffective in the P. acnes strain that responds to clindamycin.
[0038] Figs, 2A a d 2 show concentration-dependent inhibition of DNA
gyrase activity (super-coiling) by compound 91. Fig. 2A - Agarose gel
electrophoresis showing effect of compound 9 1 on super-coiling of E. coli
p as id DNA by DNA Gyrase. Fig. 2B -Percentage of DNA super-coiling by
DNA gyrase in presence increasing concentrations of compound 91.
[0039] Figs. 3A is a bar graph showing percentage of DNA super-coiling by
DNA gyrase in presence of compounds 90, 91, 94, 113, 115 and 116 with relaxed
E. coli plasmid DNA Compound 9land compound 1 6 seemed to have the best
gyrase inhibiting activity among al the comparators. Though this observation is
mostly correlated with MIC data agianst P. acnes, yet there is some speciesspecific
advantages is observed with compound 91.
[0040] Fig. 3B is a bar graph showing percentage of DNA super-coiling by
DNA gyrase in presence of compound 9 1 and nadifloxacin with relaxed E. coli
plasmid DNA. Compound 9 1 showsgreaterefficacy than Nadifloxacin.
[0041] Figs. 4A a d 4B are bar graphs showing the effect of compound 9 1 on
P. c -induced cytokine L - 6 (Fig. 4A), L - 8 (Fig. 4B) release in THP-1
cells. Compound 9 1 exerts an anti-inflammatory activity against P. c e -induced
cytokine production. Statistical analysis was performed using Student's West (* p
= 0.05; ** p = 0.005).
[0042] Figs. 5A a d 5B are bar graphs showing the effect of compound 91 on
P c -induced cytokine IL - l a (Fig. 5A), IL - 1 (Fig. SB) release in THP-1
cells.
[0043] Fig, 6, is a bar graph showing the minimum inhibition value for some
exemplary topical gel formulations against P. acnes.
[0044] Fig, 7 is a line graph showing the dose response curve of Zone of
inhibition (ZOI) of some exemplary ge formulations against P. acnes.
[0045] Fig, 8 is a line graph showing the time kill kinetics of some exemplary
gel formulations against P. acnes.
[0046] Fig, 9 Graph shows the efficacy of a topical formulation of
besifloxacin in P. acnes in an in vivo skin infection model. Besifloxacin gel
formulation has the ability to clear almost 1.5 log CFU (-95%) of inoculum of
clindamycin resistant P. acnes within first 24 hours. .
[0047] Fig. 10 is a line graph showing time kill kinetics of some exemplary
besifloxacin formulations against P. acnes MTCC 1951, showing that the
composition of the formulation can change the efficacy of an antibiotic.
[0048] Fig. His a line graph showing time kill kinetics of some exemplary
besifloxacin formulations and besifloxacin AP s against S. aureus.
[0049] Fig. 12 is a line graph showing time kill kinetics of besifloxacin
against P. acnes (CCARM 90 0)
[0050] Fig. 13 Graph shows that topical formulations with different excipient
compositions for the same antibiotic can result in different profiles in the skin and
in systemic circulation of SD rats. Fully suspended 1% Besifloxacin ge (VLNF19/
BSF/GL/068), fully soluble 1% Besifloxacin gel (VLN-F21/BSF/GL/001A)
and fully suspended 1% Besifloxacin gel (VLN-F20/BSF/CR/004) were used for
comparison purpose. To be efficacious, a formulation should not only be
physicochemically compatible with the antibiotic but also enable a sustained
concentration of the antibiotic at a concentration greater than M C level.
[0051] Figs 14A and 14 are bar graphs showing the concentrationdependent
inhibitory effect of Besifloxacin on P. c ei -induced cytokines IL-6
(Fig. 14A) but not IL-8 (Fig. 14B) release in THP-1 cells. Statistical analysis was
performed using Student's /-test (* p = 0.005; ** p = 0.0005).
[0052] Figs. SA a d 15B are bar graph showing the combination of
Besifloxacin and Adapalene increases the efficacy of inhibiting P. acnes-induced
cytokines IL-6 (Fig, SA) but has no effect on IL-8 (Fig. 15B) release in THP-1
cells. Statistical analysis was performed using Student's est (* p = 0 005; ** r =
0.001).
DETAILED DESCRIPTION OF THE INVENTION
[0053] Acne vulgaris is a skin condition that affects over 85% of a l people.
Acne is a term for a medical condition of plugged pores typically occurring on the
face, neck, and upper torso. Following are four primary factors that are currently
known to contribute to the formation of acne vulgaris; (1) increased sebum output
resulting in oily, greasy skin; (2) increased bacterial activity, normally due to an
overabundance of Propionibacterium acnes bacteria; (3) plugging
(hypercornification) of the follicle or pilosebaceous duct; and (4) and
inflammation. The plugged pores result in blackheads, whiteheads, pimples or
deeper lumps such as cysts or nodules. Severe cases of acne can result in
permanent scarring or disfiguring.
[0054] As articulated in http://thescienceofacne.com antibiotic-susceptibilitvof-
propionibacterium-acnes/ results from studies over last four decades clearly
demonstrate that over time P. acnes bacteria has become increasingly resistant to
certain classes of antibiotics.Particularly important are observations that a
significant percentage of the bacteria isolated from acne patients are now resistant
to the most common antibiotics used in acne treatment: Clindamycin,
Erythromycin, Tetracycline, Doxycycline and Minocycline. Additionally, such
resistant or antibiotic-tolerant strains can cause relapse of acne, and also cause
other diease states. There is a need for antibiotics that can kill P. acnes while
minimizing the probability of development of mutant or tolerant strains that can
survive the antibiotic exposure, and those that can work against strains that are not
responding to the current drugs. Additionally, if these novel molecules can target
additional steps in acne formation, such as inflammation, then the clinical
outcome in acne can be greater than existing therapies.
[0055] Skin is a major organ of the body, and performs many essential
functions besides acting as a barrier, such as maintaining homeostasis. Besides
acne, there are many other skin diseases that are caused by bacterial colonization
of the skin. The most common bacteria for mild to moderate skin infection are
Staphylococcus and Streptococcus e.g., Acute Bacterial Skin and Skin Structure
infection (ABSSSI). Such bacteria can infect the skin of both pediatric and adult
patients; mainly develop during hospitalization or living in a nursing home, while
gardening, or while swimming. Some people are at particular risk of developing
skin infections, for example, people suffering with diabetes, human
immunodeficiency virus (HIV) or AIDS or other immune disorders, or hepatitis,
and who is undergoing chemotherapy or treatment with other drugs that suppress
the immune system.
[0056] Common skin bacterial infections include cellulitis, erysipelas,
impetigo, folliculitis, and furuncles and carbuncles. Cellulitis is a painful,
erythematous infection of the dermis and subcutaneous tissue characterized by
warmth, edema, and advancing borders and is usually caused by Streptococcus or
Staphylococcus species. Erysipelas is a superficial form of cellulitis with sharply
demarcated borders and is caused almost exclusively by Streptococcus. Impetigo
is also caused by Streptococcus or Staphylococcus and can lead to lifting of the
stratum comeum resulting in the commonly seen bullous effect. Folliculitis is an
inflammation of the hair follicles, and it is most commonly caused by
Staphylococcus. If the infection of the follicle is deeper and involves more
follicles, it moves into the furuncle and carbuncle stages and usually requires
incision and drainage. Two different kinds of skin diseases occurred due to the
toxins produced by the bacteria include, Staphylococcal Scaled Skin Syndrome
(SSSS) which usually affects children less than 5 years old, adults with kidney
failure and the other one is Toxic Shock Syndrome.There is more chance of
colonization of S. aureus is found with patients suffering from eczema and atopic
dermatitis, a type of inflammatory, relapsing, non-contagious, itchy skin disorder.
Thus Staphylococcus aureus infection plays an important ro e in atopic dermatitis
(AD) or atopic eczema (AE). Unfortunately, some strains of Staphylococcus have
become resistant to methicillin and other similar antibiotics which are known as
MRSA. Recently it has been found that more than one-half of all cases of skin
bacterial infections caused by MRSA species. The infections associated with
MRSA species cannot be cured with traditional penicillin-related drugs. Instead,
MRSA must be treated with alternate antibiotics.
[0057 However as articulated in http://thescienceofacne.com/antibioticsuscept'ibi
¾ity-of-propionibacterium-acnes/ "Not all antibiotics are created equal".
The same is true for bacteria. Some types of antibiotics are highly effective
against certain types of bacteria, while essentially worthless against others.
Moreover, antibioitic susceptibility and resistance is a dynamic process that is
constantly changing. Over time, certain types of bacteria may gain or lose
resistance to particular antibiotics. The primary problem with standard,
laboratory-based antibiotic resistance testing is that the susceptibility of a bacteria
to an antibiotic is often different when it is growing on a petri dish versus when it
is growing on your body. This is because bacteria are not static organisms, they
adapt to their environment. A P. acnes bacteria growing in a follicle and feeding
on sebum has a different metabolic profile than one growing on a petri dish and
feeding on a bacterial nutrition supplement. Furthermore, bacteria modulate
expression of surface proteins, eel! wall structures and genesdepending on their
environment, and these changes can have a profound effect on their susceptibility
to a particular antibiotic. As a result, in the case of topical antibiotics for treatment
of skin bacterial conditions, apriori knowledge does not exist, i.e. there is no
mechanism to predict that an antibiotic will be effective against P. acnesacnes or
any other skin bacterial condition until it has been tested on the bacterial strain.
For example, as shown in http://thescienceofacne.corri/antibiotic-susceptibilitv-ofpropionibacterium-
acnes/ , P. acnes was reported to be highly resistant to a
nitroiniidazole (metronidazole) or a tetracycline (lymecycline) but partially
responsive to doxycycline (another tetracycline), and showing no resistance to
ciprofloxacin but resistant to another fluoroquinolone, Levofloxacin.lt is therefore
impossible to predict which antibiotic will work based on a priori activity in other
bacterial strains. There is a need for a systematic development of novel antibiotics
that show activity against acne.
[0058] In this regard DART molecules can act as an ideal drug candidate to
acne caused by P. acnes, and additionally for the treatment of other skin and skin
structure infections caused by other bacteria such as MRSA. DARTs were
designed to contain two distinct chemical domains, selected from different
families as mentioned earlier, for example a b-lactam ring and a quinolone
nucleus, or a quinolone nuclues and nitro-heterocyele, or a b-lactarn ring and a
nitroheterocycle, which confers two distinct mechanisms of action. This creates
less chance in mutation development at both target sites of bacteria resulting in
less resistance development against these antibiotics. Some of the molecules can
exert additional anti-inflammatory mechanisms to reduce host inflammatory
response, further enhancing the anti-acne efficacy.
[0059] The embodiments of the various aspects disclosed herein are based on
the molecules designed by the inventors, which can act on at least two different or
distinct targets. Generally, the molecule includes at least two different or distinct
chemical domains. Each of said chemical domains binds to a distinct or different
active site in a target cell. The said chemical domains can be bound together
through a third domain. As used herein, the term "chemical domain" means a part
of a molecule that is involved in a desired property. For example, a chemical
domain can be part of the molecule involved in binding of the molecule with a
target or involved in modulating an activity of the target.

REFERENCES
* Jeremy et al (2003). Journal of Investigative Dermatology; 12 : 20-27;
® Thibout et al (2014). Journal of Investigative Dermatology; 134: 307-310;
* Tagiietti e al (2008) Skin Therapy Letter. 2008; 13(5): 6-8;
Regoes et al. (2004) Antimicrob Agents Chemother:48(10): 3670-6;
* Miller et al. (2004) Science;305 (5690): 629-3 1;
* Keren et al. (2004) J. Bacterid; 186: 8172-8180;
* Schulzen e al. (2001), Nature 4 3: 814-821;
* Beitru et al. (2003) Antimicrob Agents Chemother.; 47(3): 1 12-1 4;
® Cambau et al. (2009). Antimicrob. Chemother.63 (3): 443-450;
* Kim et al. (2002) Journal of immunology; 169(3): 1535-41;
* Liu et al.(2005) Journal of Immunology; 174(5): 2467-2470;
* Nagy et al.(2006) Microbes infect; 8(8): 2 195-205;
* Lee et al. (2010) Arch Dermatol Res; 302(10): 745-56;
* Mouser et a . (2003) J nvest Dermatol; 121(5): 1226-8;
* Zasloff et a (2002) Nature; 415: 389-395;
® Epand et al. (1999) Biochim Biophys Acta: 1 62: 1-28;
Kabara e al. (1972) Antimicrobn Agents Chemother; 2(1): 23-28; and
* De Lucca et ai. (2000) Rev. Iberoam. Micol; 17: 1 6-120
2 16
The formulation of claim 1, wherein the formulation is formulated for topical,
oral or parenteral administration.
The formulation of claim 10, wherein the formulation is an oral dosage,
injectable, aerosol or inhalant, lotion, cream, gel, emulgel, oil, serum, powder,
spray, ointment, solution, suspension, dispersion, paste, foam, peel, films, mask,
patch, stick, roller, impregnated fabric, or any combination thereof.
The formulation of claim 1, further comprising a second anti-baterial agent.
The formulation of claim 12, wherein the second anti-baterial agent t is selected
from the group consisting of 8-chloro fluroquinolones, acetretin, adapalene(s),
alitretinoin, alpha- or beta-hydroxy acids, antibiotics, antimicrobial peptides,
antimicrobials, azelaic acid, benzoyl peroxide, bexarotene, bile salts, biofilm
inhibitors, clindamycin, erythromycin, etretinate, glycolic acid, isotretinoin,
keratolytic agents, lactic acid, lipoic acid, N-acetylcystein, natural anti-acne
agents, octopirox, phenoxyethanol, phenoxypropanol, pyruvic acid, resorcinol,
retinoic acid, retinoid(s), salicylic acid, sebostats, sodium sulfacetamide,
spironolactone, sulfur, sulfur containing D- or L-amino acids, tazarotene, tea tree
oil, tretinoin, triclosan, urea, and any combinations thereof.
The formulation of claim 1, wherein the formulation comprises an 8-chloro
fluoroquinolone alone or in combination with a second anti-baterial agent.
The formulation of claim 14, wherein the formulation comprises besifloxacin and
adapalene.
The formulation of claim 14, wherein the formulation comprises 8-
chlorofluoroquinolone and an anti-inflammatory agent.
The formulation of claim 16, wherein the formulation comprises 8-
chlorofluoroquinolone and retinoic acid or retinoid.
The formulation of claim 12, wherein the first or the second anti-bacterial agent
is in the form of a drug carrier comprising the said first or second anti-bacterial
agent and at least one additional compound, said additional compound selected
from the group consisting of lipids, oils, polymers, peptides, proteins,
carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and
any combinations thereof.
The formulation of claim 8, wherein the second anti-bacterial agent drug carrier
has a size of about 5 nm to about 50 m h.
218
The formulation of claim 19, wherein the second anti-bacterial agent drug carrier
has a size of about 100 nm to about 25 mpi
The formulation of claim 18, whereinthe second anti-bacterial agent drug carrier
comprises a surface modifier on the surface thereof.
The formulation of claim 19, wherein the surface modifier of the second anti¬
acne agent drug carrier is selected from the group consisting of lipids, oils,
polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids,
lipoproteins, cationic molecules, and any combinations thereof.
The formulation of claim 18, wherein the surface of the second anti-bacterial
agent drug carrier is substantially free of surface modifier.
The formulation of claim 1, further comprising an additional active agent.
The formulation of claim 24, wherein the additional active agent is an antiinflammatory-
agent, penetration enhancer, anti-oxidant, anti-aging agent, antiwrinkle
agent, skin whitening or bleaching agent, ultraviolet (UV) light absorbing
or scattering agent, skin depigmentation agent, skin regenerative agent, scar
healing agent, or any combination thereof
The formulation of claim 24, wherein the additional active agent is in the form of
a drug carrier comprising a compound selected from the group consisting of
lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids,
phospholipids, lipoproteins, cationic molecules, and any combinations thereof.
The formulation of claim 26, wherein the additional active agent drug carrier has
a size of about 5 nm to about 100 mh .
The formulation of claim 27, wherein the additional active agent drug carrier has
a size of about 100 nm to about 25 m h .
The formulation of claim 24, whereinthe additional active agent drug carrier
comprises a surface modifier on the surface thereof.
The formulation of claim 29, wherein the surface modifier of the additional active
agent drug carrier is selected from the group consisting of lipids, oils, polymers,
peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins,
cationic molecules, and any combinations thereof.
The formulation of claim 24, wherein the surface of the additional active agent
drug carrier is substantially free of surface modifier.
The formulation of claim 1, wherein the formulation further comprises a zinc
compound.
219
33. The formulation of claim 33, wherein the formulation comprises a moisturizing
agent, humectant and /or emolient.
34. The formulation of claim 1, wherein the formulation comprises an antibiotic in
soluble form.
35. The fomulation as claimed in any preceding claim having the following
composiiton:
36. Use of 8-chloro fluoroquinolone for the treatment of acne, particularly, drug
resistant acne.
37. Use of 8-chloro fluoroquinolone for the preparation of medicament for the
treatment of acne, particularly, drug resistant acne.
38. Use of besifloxacin for the treatment of acne, particularly, drug-resistant acne.
39. Use of besifloxacin for the treatment of acne via resolution of inflammation.
220
Use of besifloxacin, either alone or in combination with another active for the
preparation of medicament for the treatment of acne, particularly, drug-resistant
acne.
A method for treatment of acne, which comprises administering to a patient in
need thereof an effective amount of a formulation as claimed in any one of claims
1 to 35.
A method for treatment of acne, particularly where P acne is present and is not
responding to a therapeutic dose of clindamycin, minocycline, tetracycline and
erythromycin, which comprises administering to a patient in need thereof an
effective amount of a formulation as claimed in any one of claims 1 to 35.
A Dual Action Rational Therapeutic (DART) molecule that has two distinct
mechanisms of action for treatment or prevention of bacterial infections.
A Dual Action Rational Therapeutic (DART) molecule of claim 43 that has
two distinct anti-bacterial mechanisms of action or an anti-bacterial and an
anti-inflammatory action.
The molecule of claim 43, wherein the molecule inhibits DNA gyrase or
topoisomerase IV and transpeptidase-mediated cross-linking of peptidoglycans.
The molecule of claim 43, wherein the molecule inhibits isoprenyl pyrophosphate
and transpeptidase-mediated cross-linking of peptidoglycans.
The molecule of claim 43, wherein the molecule inhibits isoprenyl pyrophosphate
and DNA gyrase of topoisomerase IV.
The molecule of claim 43, wherein the molecule inhibits folate synthesis and
DNA gyrase of topoisomerase IV.
The molecule of claim 43, wherein the molecule inhibits folate synthesis and
transpeptidase-mediated cross-linking of peptidoglycans.
The molecule of claim 43, wherein the molecule inhibits DNA gyrase or
topoisomerase IV and the 30S sub-unit in bacteria.
The molecule of claim 43, wherein the molecule inhibits DNA gyrase or
topoisomerase IV and the 50S sub-unit in bacteria.
The molecule of claim 43, wherein the molecule inhibits transpeptidase-mediated
cross-linking of peptidoglycans and the 30S or the 50S sub-unit in bacteria.
The molecule of claim 43, wherein the molecule inhibits folate synthesis and the
30S or the 50S sub-unit in bacteria.
221
The molecule of claim 43, wherein the molecule inhibits isoprenyl pyrophosphate
and the 30S or the 50S sub-unit in bacteria.
The molecule of claim 43, wherein the molecule exerts an antimicrobial activity
via the generation of toxic intermediates and/or free radicals while additionally
inhibiting DNA gyrase and/or topoisomerase.
A Dual Action Rational Therapeutic (DART) molecule that has two distinct anti¬
acne mechanisms of action.
The molecule of claim 56, wherein the molecule modulates at least two different
targets in acne.
The molecule of claim 56, wherein the molecule modulates at least two different
targets in P. acnes.
The molecule of claim 56, wherein the first mechanism is an antibacterial action
and the second mechanism of action is inhibition of keratinocyte proliferation
and differentiation.
The molecule of claim 56, wherein the first mechanism is an antibacterial action
and the second mechanism of action is anti-inflammatory.
A Dual Action Rational Therapeutic (DART) molecule which comprises a
quinolone and a nitro-heterocycle.
A Dual Action Rational Therapeutic (DART) molecule that comprises a betalactam
and a nitro-heterocycle.
A Dual Action Rational Therapeutic (DART) molecule that comprises a betalactam
and a quinolone.
A Dual Action Rational Therapeutic (DART) molecule which includes two
chemical domains, each said chemical domain binding to a distinct active site in
target cells, wherein said chemical domains are bound together through a third
domain.
The molecule of claim 64, wherein the third domain is a linker.
The molecule of claim 64, wherein the third domain is a cleavable linker.
The molecule of claim 64, wherein the third domain is a non-cleavable linker.
The molecule of claim 64, wherein said third domain is 11-hydroxyundecenic
acid; 1,10-decanediol; 1,3 -propanediol; 1,5-pentanedil; 10-hydroxydecenic acid;
succinic; lactic acid; 3-hydroxypropionic acid; or any combination thereof..
The molecule of claim 64, wherein the third domain increases an activity of at
least one of the two chemical domains.
222
The molecule of claim 64, wherein the third domain has antibacterial or anti¬
inflammatory activity.
The molecule of any of claims 43-70, wherein the molecule is in the form of a
drug carrier.
The molecule of any of claims 43-70, wherein the molecule is in solubilized
form.
The molecule of claim 71, wherein the drug carrier has a size of about 5 m to
about 100 m
The molecule of claim 71, wherein the drug carrier has a size of about 100 n to
about 25 m .
The molecule of claim 71, wherein the drug carrier further comprises a
compound selected from the group consisting of lipids, oils, polymers, peptides,
proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic
molecules, and any combinations thereof.
The molecule of claim 71, wherein the drug carrier further comprises an
additional active agent.
The molecule of claim 71, wherein the additional active agent is an antiinflammatory-
agent, keratolytic agent, penetration enhancer, anti-oxidant, antiaging
agent, anti-wrinkle agent, skin whitening or bleaching agent, ultraviolet
(UV) light absorbing or scattering agent, skin depigmentation agent, skin
regenerative agent, scar healing agent, or any combination thereof.
The molecule of claim 71, wherein surface of the drug carrier is substantially free
of surface modifier.
The molecule of claim 71, wherein the drug carrier further comprises an
additional anti-acne agent.
The molecule of claim 79, wherein the second anti-acne agent is selected from
the group consisting of acetretin, adapalene(s), alitretinoin, alpha- or betahydroxy
acids, antibiotics, antimicrobial peptides, antimicrobials, azelaic acid,
benzoyl peroxide, bexarotene, bile salts, biofilm inhibitors, clindamycin,
erythromycin, etretinate, glycolic acid, isotretinoin, keratolytic agents, lactic acid,
lipoic acid, N-acetylcystein, natural anti-acne agents, octopirox, phenoxyethanol,
phenoxypropanol, pyruvic acid, resorcinol, retinoic acid, retinoid(s), salicylic
acid, sebostats, sodium sulfacetamide, spironolactone, sulfur, sulfur containing
223
D- or L-amino acids, tazarotene, tea tree oil, tretinoin, triclosan, urea, and any
combinations thereof.
81. The molecule of claim 71, wherein the drug carrier further comprises a surface
modifier on the surface thereof.
82. The molecule of claim 81, wherein the surface modifier is a compound selected
from the group consisting of lipids, oils, polymers, peptides, proteins,
carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and
any combination thereof.
83 A DART molecule as re resented by the structure:
DART 90
84. A DART molecule as represented by the structure:
DART 9 1
85 A DART molecule as represented by the structure:
DART 115
224
A DART molecule as represented by the structure:
DART 116
The molecule as claimed in any one of the claims 43-82 is selected from
structures shown in claims 83-86
The formulation comprising a dual action rational therapeutic molecule of any of
claims 43-87 and at least one carrier or excipient.
The formulation of claim 88, wherein the carrier or excipient is selected from the
group consisting of emulsifiers, preservatives, surfactants, oils, lipids, waxes,
stabilizers, rheology modifiers or thickening agents (gelling agent), emollients,
moisturizers, conditioning agents, fragrances/perfumes, potentiating agents,
preservatives, opacifiers, antioxidants, cooling agents, film forming agents,
abrasives, exfoliating agents, colorants, pH modifiers, solvents, vehicle,
penetration enhancers, permeation enhancers, pearlizing agents, and any
combinations thereof.
The formulation of claim 88 comprising from about 5% to about 99% (w/w or
w/v) of the carrier or excipient.
The formulation of claim 88, wherein the formulation is formulated for topical,
oral or parenteral administration.
The formulation of claim 88, wherein the formulation is an oral dosage,
injectable, aerosol or inhalant, lotion, cream, gel, emulgel, oil, serum, powder,
spray, ointment, solution, suspension, dispersion, paste, foam, peel, films, mask,
patch, stick, roller, impregnated fabric, or any combination thereof.
The formulation of claim 88 further comprising a second anti-acne agent.
The formulation of claim 91, wherein the second anti-acne agent is selected from
the group consisting of acetretin, adapalene(s), alitretinoin, alpha- or betahydroxy
acids, antibiotics, antimicrobial peptides, antimicrobials, azelaic acid,
benzoyl peroxide, bexarotene, bile salts, biofilm inhibitors, clindamycin,
erythromycin, etretinate, glycolic acid, isotretinoin, keratolytic agents, lactic acid,
lipoic acid, N-acetylcystein, natural anti-acne agents, octopirox, phenoxyethanol,
225
phenoxypropanol, pyruvic acid, resorcinol, retinoic acid, retinoid(s), salicylic
acid, sebostats, sodium sulfacetamide, spironolactone, sulfur, sulfur containing
D- or L-amino acids, tazarotene, tea tree oil, tretinoin, triclosan, urea, and any
combinations thereof.
95. The formulation of claim 91, wherein the second anti-acne agent is in the form of
a drug carrier.
96. The formulation of claim 93, wherein the second anti-acne agent drug carrier
further comprises a compound selected from the group consisting of lipids, oils,
polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids,
lipoproteins, cationic molecules, and any combinations thereof.
97. The formulation of claim 93, wherein the second anti-acne agent drug carrier has
a size of about 5 nm to about 100 m h .
98. The formulation of claim 95, wherein the second anti-acne agent drug carrier has
a size of about 100 nm to about 25 m h.
99. The formulation of claim 93, wherein the second anti-acne agent drug carrier
comprises a surface modifier on the surface thereof.
100. The formulation of claim 97, herein the surface modifier of the second anti-acne
agent drug carrier is selected from the group consisting of lipids, oils, polymers,
peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins,
cationic molecules, and any combinations thereof.
101. The formulation of claim 93, wherein surface of the second anti-acne agent is
substantially free of surface modifier.
02. The formulation of claim 88 further comprising an additional active agent.
103. The formulation of claim 100, wherein the additional active agent is an antiinflammatory-
agent, penetration enhancer, anti-oxidant, anti-aging agent, antiwrinkle
agent, skin whitening or bleaching agent, ultraviolet (UV) light absorbing
or scattering agent, skin depigmentation agent, skin regenerative agent, scar
healing agent, or any combination thereof.
104. The formulation of claim 100, wherein the additional active agent is in the form
of a drug carrier.
105. The formulation of claim 102, wherein the additional active agent drug carrier
further comprises a compound selected from the group consisting of lipids, oils,
polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids,
lipoproteins, cationic molecules, and any combinations thereof.
226
A formulation for use in the treatment or prevention of acne, including drugresistant
acne and P.acne not responding to/tolerant to a therapeutic dose of
clindamycin, minocycline, tetracycline or erythromycin, comprising an anti
bacterial agent and at least one additional compound, selected from a carrier,
excipient lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids,
phospholipids, lipoproteins, cationic molecules, and any combinations thereof.
The formulation of claim 1, wherein the drug carrier comprises the said anti¬
bacterial agent uncoated or coated at least partially with said additional
compound.
The formulation of claim 2, wherein the anti-baterial agent has a size of about 5
nm to about 20 m .
The formulation of claim 3, wherein the drug carrier has a size of about 50 nm to
about 10 m .
The formulation of claim 1 further comprising a surface modifier on the surface
of the anti-baterial agent.
The formulation of claim 5, wherein the surface modifier is selected from the
group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates,
glycolipids, phospholipids, lipoproteins, cationic molecules, and any
combinations thereof.
The formulation of any of claims 1-6, wherein the carrier or excipient is selected
from the group consisting of emulsifiers, preservatives, surfactants, oils, lipids,
waxes, stabilizers, rheology modifiers or thickening agents (gelling agent),
emollients, moisturizers, conditioning agents, fragrances/perfumes, potentiating
agents, preservatives, opacifiers, antioxidants, cooling agents, film forming
agents, abrasives, exfoliating agents, colorants, pH modifiers, solvents, vehicle,
penetration enhancers, pearlizing agents, and any combinations thereof.
The formulation of claim 1, wherein the surface of the anti-baterial agent is
substantially free of surface modifier.
The formulation of claim 1, comprising from about 0.1% to about 50% (w/w or
w/v) of the carrier or excipient.
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106. The formulation of claim 102, wherein the additional active agent drug carrier
has a size of about 5 n to about 00 m .
107. The formulation of claim 104, wherein the additional active agent drug carrier
has a size of about 00 nm to about 25 m .
108. The formulation of claim 102, wherein the additional active agent drug carrier
comprises a surface modifier on the surface thereof.
109. The formulation of claim 106, wherein the surface modifier of the additional
active agent drug carrier is selected from the group consisting of lipids, oils,
polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids,
lipoproteins, cationic molecules, and any combinations thereof.
110. The formulation of claim 102, wherein surface of the additional active agent drug
carrier is substantially free of surface modifier.
11 . The formulation of claim 88, wherein the formulation further comprises a zinc
compound.
112. A formulation comprising a dual action rational therapeutic molecule claimed in
any of claims 83-86 and at least one carrier or excipient.
113. A formulation comprising a dual action rational therapeutic molecule claimed in
any of claims 83-86 and at least one carrier or excipient for topical application.
114. A formulation comprising a dual action rational therapeutic molecule claimed in
any of claims 83-86 and at least one carrier or excipient for topical application on
skin.
115. Use of Dual Action Rational Therapeutic (DART) molecule for the treatment of
acne.
116. Use of Dual Action Rational Therapeutic (DART) molecule for the treatment
drug-resistant acne.
117. Use of Dual Action Rational Therapeutic (DART) molecule for the preparation of
medicament for the treatment of acne, particularly, drug-resistant acne.
18. Use of Dual Action Rational Therapeutic (DART) molecule for the treatment of
acne, particularly, drug resistant-acne.
119. Use of Dual Action Rational Therapeutic (DART) molecule, either alone or in
combination with another active for the preparation of medicament for the
treatment of acne, particularly, drug-resistant acne.
120. Use of Dual Action Rational Therapeutic (DART) molecule, either alone or in
combination with another active for the preparation of medicament for the
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treatment of acne, particularly, P. acnes not responding to clindamycin,
minocycline, tetracycline and erythromycin-containing treatments.
2 . A method for treatment of acne, particularly, acne not responding to clindamycin,
erythromycin, minocycline, doxycycline or tetracycline-containing therapies,
which comprises administering to a patient in need thereof an effective amount of
a formulation as claimed in any one of claims 88-1 14.
122. The method of any of claim 42 or 121, wherein the acne condition is caused by
antibiotic-susceptible bacterial strain.
123. The method of claim 42 or 121, wherein the acne condition is caused by bacteria
not responding to therapeutic doses of clindamycin-, erythromycin-, doxycycline-
, minocycline- or tetracycline-containing regimens.
124. The method of claim 123, wherein the acne condition is caused by clindamycin-,
tetracycline-, minocycline-, doxycycline-, or erythromycin-nonresponder
Propionbacterium acnes.
125. The method of claim 123, wherein the acne condition is caused by clindamycin-,
tetracycline-, minocycline-, doxycycline-, or erythromycin-tolerant
Propionbacterium acnes.
126. A method of treating a bacterial infection in a subject comprising administering a
therapeutically effective amount of a formulation of any of claims 1-35 and 88-
114.
127. The method of claim 126, wherein the infection is caused by a pathogen selected
from the group consisting of Bartonella henselae, Borrelia burgdorferi,
Campylobacter jejuni, Campylobacterfetus, Chlamydia trachomatis, Chlamydia
pneumoniae, Chylamydia psittaci, Simkania negevensis, Escherichia coli (e.g.,
0157:H7 and K88), Ehrlichia chafeensis, Clostridium botulinum, Clostridium
perjringens, Clostridium tetani, Enterococcus faecalis, Haemophilius influenzae,
Haemophilius ducreyi, Coccidioides immitis, Bordetella pertussis, Coxiella
burnetii, Ureaplasma urealyticum, Mycoplasma genitalium, Trichomatis
vaginalis, Helicobacter pylori, Helicobacter hepaticus, Legionella pneumophila,
Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium ajricanum,
Mycobacterium leprae, Mycobacterium asiaticum, Mycobacterium avium,
Mycobacterium celatum, Mycobacterium celonae, Mycobacterium fortuitum,
Mycobacterium genavense, Mycobacterium haemophilum, Mycobacterium
intracellulare, Mycobacterium kansasii, Mycobacterium malmoense,
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Mycobacterium marinum, Mycobacterium scrofulaceum, Mycobacterium simiae,
Mycobacterium szulgai, Mycobacterium ulcerans, Mycobacterium xenopi,
Corynebacterium diptheriae, Rhodococcus equi, Rickettsia aeschlimannii,