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BIOD :LIVERY SYSTEMS
11' ieki of the Invention
[0001] The field of the invention generally relates to hiodelÃve.Ã r: systems
for
pro siding products or compounds, such as chemicals, to industrial systems,
The
invention also .relates to compositions for use in a. targeted delivery of
said compositions
to bacterial biofllms various environments.
Background of the invention
[0002] l3actefl'al biofilms exist in natural, medical, and industrial
environments.
The biofilms offer a selective advantage to microorganisms to ensure the
microorganisms' survival or to allow them a certain time to exist in a dormant
state until
suitable growth conditions arise. infortunately, this selective advantage
poses serious
threat's to heaaltla, or to the e.{ :iciency and lifetime of: industrial
systems. The biofil.ms must
be minimized or destroyed to improve the efficiency of industrial systems, or
remove the
potential health threats.
[0003] I I:a7.ny industrial or commercial operations rely on large quantities
of x~w,ater
for various reasons, such as for cooling systems, or said systems may produce
large
quantities ofwaÃstec-ater, which result in the creation of biohlms that need
to be treated.
These industries include, but are not limited to, a ;riceÃlture, petroleum,
oil drilling oil
pipelines, oil. storage. gas drilling, gas pipelines. gas storage. chemical,
pharmaceutical,
mining, metal plating, textile, papermakin4g, brewing, food and beverage
processing, and
semiconductor industries. In these operations, naturally occurring biof lmÃs
are
continuously produced and often accumulate on numerous structural or equipment
surfaces or on natural or biological surfaces. In industrial settings, the
presence of these
biofilms causes a decrease in the efficiency of industrial machiner-,,
requires increased
maintenance and presents potential health hazards, An example is the surfaces
of water
cooling towers which become increasingly coated with inicrobially produced
bio.fllin
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slime Which constricts Water flow and reduces heat exchange capacity
Specifically' in
flowing or stagnant water, biollms can cause serious problems, including
pipeline
blockages, corrosion of equipment by growth of underfiln n ricrobes and the
growth of
potentially harmful pathogenic bacteria. Water cooling tower biofilms may form
a
harbor or reservoir that perpetuates growth. of pathogenic nricroorganisi s
such as
Legioncllca pica anolpl ilea.
[0004] Another example of industrial systems are those systems that are found
in
the .food and beverage industries. Food preparation lines are routinely
plagued by blofilni
build-up both on the machinery and on the food product where bioflms often
include
potential pathogens, Industrial biofilmrs, such as those found in the food
industry, are
complex assemblages of insoluble polysaccharide-rich hiopolyrmers, which are
produced
and elaborated by surface dwelling micrroorganisms. More particularly,
biofilms or
microbial slimes are composed of polysaccharides, proteins and
lipopolysaccharides
extruded from certain microbes that allow them to adhere to solid surfaces in
contact with
water environments and form persistent colonies of sessile bacteria that
thrive within a
protective film. The film may allow anaerobic species to gro'~w~, producing
acidic or
corrosive conditions. To control these problems, processes and rntirnicrobial
products
are needed to control the formation and growth ofbiofilms. Control ofbiof lms
involves
the prevention of microbial attachment and/or the removal of existing
b:ic.f:ilms from
surfaces. While removal in many contexts is accomplished by short cleansing
treatments
With highly caustic or oxidizing agents the most common l y used mater ials to
control
biof lms are biocides and dispersants, In t S. patent no. 5,4I 1.,666 a method
of removing
a biofilm or preventing buildup of a biollm on a solid substrate is taught,
that comprises
a combination of at least two hiologicallv produced enzymes, such as an acidic
or
alkaline protease and a glucoarnylase or alpha amylase and at least one
surfactant. U. S.
patent no. 6,759,040 teaches a method for preparing biofrlm degrading,
multiple
specificity, hydrolytic enzyme mixtures that are targeted to remove specific
biofiirns.
[0006] U.S. patent no. 6,267 89, relates to a method of inhibiting l iotilm
formation in commercial and industrial water systems by adding one or more
plant oils to
the system. However., although the biocides are e.1edive in controlling
dispersed
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microorganism suspensions, i.e. planktonic microbes-, biocides do :riot work
well a against
sessile microbes, the basis of biolrars. This is due to the fact that biocides
have difficulty
penetrating the poivsacchande/protein slime layers surrounding the microbial l
cells.
Thicker biofilms see little penetration of biocides and poor biocide efficacy,
is the result.
One known. method of trying to better control biof-ilms has been the addition
of
dispersants and wetting agents to biocide compositions to enhance biocide
eflica.cy.
Biodispersants r ray= operate to keep planktonic microbes sufficiently
dispersed so that
they do not agglomerate or achieve the local densities necessary to initiate
the
extracellular processes responsible for anchoring to a surface, or initiating
film- or
colony,-torrrrirrgmechanisms As components in biocidal treatment formulations,
these
biodispersants have helped in opening channels in the.. biofilm to allow
better
permeability ofthe toxic agents and to better disperse the microbial
aggregates and
clumps that have been ~w~eakened and released from the surfaces. However,
biodispersants have proven to be more effective in preventing initial
biofil:.m formation
than in removing existing biofilms. In many cases, the activity of
biodispersants has been
responsible for only ?5 to 30% biornass removal from biofouled surfaces, even
when
used in conjunction with a biocida.l agent.
[0006] Therefore, a clear need still exists for an efficient and effective
means for
delivering antimicrobial compounds that are better able to penetrate existing
bioh Ims and
biofilm matrices, and more effective in killing microorganisms contained
within a biofilm
matrix, thus killing and eliminating bicf'tlm, as well as preventing future
formation nor-
buildup ofbiofilm, in systems, such as industrial systems. Decreasing the
fouling of
rnicroiltration sy=stems, and providing less frequent cleaning and/or
replacement which.
would enhance the overall frlt.ration process. are also needs which should be
addressed.
Summary of the Invention
[0007] A biodeliver, system has been found which increases the efficiency and
effectiveness of introducing antimicrobial compounds into complex biofilm
matrices,
through the use of liposome carriers, which can be used in natural, medical
and industrial
applications. hi industrial application,,,, the delivery system can minimize
or eliminate
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fouling in industrial sy sterns, including, but not limited to. aqueous
systems, such as
piping, heat exchangers, condensers, filtration systems and media, and fluid
storage
tanks.
[0008] According to one embodiment of the invention, liposorrres containing an
antimicrobial agent, such as a hydrophilic biocide, are added to a water
system prone to
biofouling and bioilrrr formation. The liposonaes, being similar in
composition to the
outer surface of the microbial cell wall structure or to the material on which
the microbes
feed, are readily incorporated into the microbes present in the existing biof
(rn. Once the
liposornes become entrained with the biofilm matrix, digestion, decomposition
or
degradation of the l peascarrre proceeds, releasing the ant microbial agent,
or biocidal
aqueous core reacts locally with the biofilm- encased microorganisms. 1. Iron
the death of
the organisms, the polysa cch,rricle?,protein matrix cannot be replenished and
decomposes
and thereby results in reduced bin fouling of the water bearing system.
Depending on the
particular system involved, this bioflnr removal or destruction therefore
results in
increased heat transfer (industrial heat exchanger), increased flux (filter or
filtration
membrane), less deposit of colloidal and particulate solids and dissolved
organics on the
surface of the rnicrofiltra.tion membrane, thereby reducing the frequency and
duration of
the membrane cleaning and ultimate replacement, or general reduction of
corrosive
surface conditions in pipelines, tanks, vessels or other industrial equipment-
[0009] An alternate embodiment of the invention provides for a delivery system
of actives into a natural, medical or industrial system, which can be chosen
from the
group consisting o.f'anti-cor-.rosion treatments, pesticides for agriculture
and commercial
home uses, food additives and preservatives, chemical and biological
detection, color and
flavor enhancement, odor control and aquatic pest managerraent.
[0010] The various features of.noveltvr that characterize the invention are
pointed
out with particularity in the claims annexed to and forming a part of this
disclosure. For a
better understanding of the invent. on, its operating advantages and benefits
obtained by
its uses, reference is made to the accompanying drawings and descriptive
matter. The
accompany ing draaxv,ings are intended to show examples of the invention The
drawings
are not intended as showing the limits of all of the ways the invention can be
made and
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used. Changes to and substitutions of the various components of the invention
can of
course be rnaade. The invention resides as well in sub-combinations and sub-
systems of
the elements described, and in methods of using them.
Brief Description of the Drawings
[0011] Refer now to the figures, which are meant to be exemplar, and not
l:imitirrw, and wherein like elements are numbered alike, and not all numbers
are repeated
in every figure for clant-y of the illustration.
[0012] Fig. I is chart setting forth results obtained from an isothiazolin
according
to one embodiment of the invention,
[0013] H& 2 is chart setting forth further results obtained from an
isothiazolin
according to one embodiment of the invention.
[0014] Fig. 3 is chart setting forth results obtained from an isothiazolin
according
to one embodiment of the invention.
[0015] pig 4 is chart setting forth results obtained from a substituted
nitnlopropionamide according to one emb xli r .meat of the inventory.
[0016] Figs. 5 and 6 are charts setting .forth results obtained from an
amrrroniurn
salt according to embodiments of the invention.
[0017] Figs. 7 and are charts setting forth results obtained from a
substituted
propanediol biocide according to embodiments of the invention.
[0018] Fig. 9 .is chart settin4g forth results obtained from a phosphonium
salt
according to one embodiment of the invention.
Detailed Description of the Invention
[0019] Approximating language, as used herein throughout the specification and
claims, may be applied to modify any quantitative representation that could
permissibly
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vary without resulting in a. change in the basic .function to which it is
related.
Accordingly, a value modified by a term or terms, such as "aboutf", is not
limited to the
precise value specified. In at least some instances, the approximating
language may
correspond to the precision of an instrument for measuring the value. Range
limitations
may be combined andior interchanged, and such ranges are identified and
include all the
sub-ranges included herein unless context or language indicates otherwise.
Other than in
the operating examples or where otherwise indicated, all numbers or
expressions
referring to quantities of ingredients, reaction conditions and the like, used
in the
specification and the claims, are to be understood as modified in all
instances by the ter n
;about".
[0020] As used herein, the terms "comprises," "comprising," "includes,
"including," "has," "having' or any other var.ation thereof, are intended to
cover a non-
exclusive inclusion. For example, a process, method, article or apparatus that
comprises
a list of elements is not necessarily limited to only those elements, but may
include other
elements not expressly listed or inherent to such process, .,Method article or
apparatus.
L0021] A. delivery system has been found which increases the efficiency and
effectiveness of introducing antimicrobial compounds into complex bicafrlm
matrices
through the use of liposo.. me carriers. which can. be used in natural,
medical and industrial
applications. Ill. industrial. applications, the delivery system can minimize
or eliminate
fouling in industrial systems, including, but not limited to, aqueous systems,
such as
cooling towers, piping, heat exclangers, condensers, filtration systems and
media, and
fluid storage tanks.
[0022] According to one embodiment of the invention, liposo..mes containing a
biocidal or antimicrobial agent or compound are added to an industrial. system
prone to
biofouling and biofilna formation_ The liposomes, being similar in
conapositic:.n to
microbial membranes or cells, are readily incorporated into the existing
biofilna. Once
the antic microbial c ampound-containing liposora-les diffuse into, adsorb or
otherwise
become entrained with the biofilm matrix, the microorganisms existing within
the biofilm
matrix will ingest the, liposorte structure, resulting in the decomposition or
disintegration
of the l:iposome inside the intracellular matrix of the .rmmicroorganis.m,
thereby releasing the
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antinnic:robial compound into the intracellular rnrat ix of the
microorganisar.r., ultimately
resulting; in the death of the microorganism. That is lipid decomposition and
biocide
release can be programmed to occur by making the lipid matrix sensitive to pl-
1, redox
potential. Ca concentration, or other changes. Thereafter the biocidal
component that
may be concentrated in the aqueous core of the liposome or in the lipid
membrane
portion of the liposome, is released to react directly with the biofilm-
encased
nmicroor<ganismms Thus, rather than adding a biocide at high levels to the
bulk water
system, a small quantity of liposome-encased biocide is taken rip by the
biofil.rr or by free
(planktonic) organisms, and degradation of the liposome releases the biocide
locally in or
at the target organisms or their film matrix .niche. The biocide thus attains
a high
concentration locally to kill the target organisms, and upon the death of the
organisms,
the polysaccharide/protein matrix that forms the biofilrrr cannot be
maintained OF
regenerated and decomposes, and thereby results in reduced fouling of the
water bearing
system, resulting in increased heat transfer., increased flux, less deposit of
colloidal and
particulate solids and dissolved organics on the surface of the
microfiltration membrane:
thereby reducing the frequency and duration of the membrane cleaning and
ultimate
replacement or other benefits.
[0023] Liposornes, or lipid bodies, are systems in which lipids are added to
an
aqueous buffer to forma vesicles, structures that enclose a volume. The l
poso:rrres may be
comprised of lipids selected from the group consisting of phospholipids,
lcthicin,
phosphatidyl choline, glycolipid, tfli<glyceride, sterol, fatty acid,
sphingolipid, or
combinations thereof.
[0024] More specifically; liposomes are microscopic vesicles, most commonly
composed of phospholipids and water. The liposorares may be made from
phospholipids
derived from various sources, including, but not limited to soybeans and eggs.
When
properly mixed, the phospholipids arrange themselves into a bilayer or multi
layers, very
similar to a cell membrane, surrounding an aqueous volume core. l_ iposernyes
can be
produced to carry various compounds or chemicals Within the aqueous cores or
the
desired compounds can be formulated in a suitable carrier to enter the lipid
layer(s).
Liposomes can be produced in various sizes and may be manufactured in
submicron to
multiple micron diameters. The Iiposc.mes may be manufactured by several known
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processes. Such processes include, but are not limited to, controlled
evaporation,
extrusion, injection, microfluid processors and rotor-stator mixers. Liposomes
can be
produced in diameters ranging from about 10 naanomete:rs to greater than
about. 1.5
micrometers. When produced in sizes from about 100 manometers to about 2
micrometer
sizes the liposomes are very similar in size and composition to most
microbial. cells. The
biocide or antimicrobial compound contai.ni.ng-hposomes should be produced in
sizes that
mimic bacterial cells, for example, from about 005 to about 15 l,t, or
alternately, about
0. 1 to 10.0 p..
[0025] In one embodiment, effective amounts of the biocide containing liposome
is introduced into an industrial system. which is prone to biof'ouling and
biofilm.
formation, or can be introduced into systems drat already exhibit signs of
biofouling or
biofilm formation. The effective amount will vary according to the
antimicrobial
compound or biocide, and the aqueous system to which it is added, but one
embodiment
provides from about 0.01 ppm to about 100 ppm, with an alternative of from
about 0.05
to about 50 ppm, alternately from about 0.05 to about 5.0 The l.iposomes,
being similar in
composition to microbial membranes, or cell walls, are readily incorporated
into the
existing biofÃlm and become entrained within the biofÃlm .matrix. Tae
liposomes
containing biocides have improved penetration of the biofilm matrix, due to
similarity in
composition acrd str .rcture with tyre hiof'rlm. Once the liposomare :is
incorporated o.r
entrained within the existing biofilm matrix, the liposome will begin to
disintegrate.
Upon the deco decomposition or programmed disintegration of the liposonae, the
biocidal
compound contained within the aqueous core of the liposome is released to
react directly
with the biofilna encased microorganisms, resulting in their demise. Upon the
death of
t:he or gani swim s, the poly sa .achari de/protei ra matrix will rapidly
decompose, treei n g the
surface from contaminating microbes.
[0026] A principal feature of one embodiment of the present invention is that
the
liposonies constitute extremely small ydropbobic'bodies that may readily
survive in and
disperse in systems, such as for example, aqueous or natural systems, and yet
will adsorb
to or penetrate a hiofi [in and preferentially target or be targeted by the
microbes that
inhabit, constitute or sustain the biofilm. As such, the liposomes deliver a
biocidaal. agent
directly to the microbes or biof lmra, resulting in effective locally biocidal
level of activity,
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without requiring that the industrial system as a. whole sustain a. high dose
Thus, here
conventional bioflm treatment may require dosing with a bulb biocidal chemical
at a
certain level, delivery via liposome may be dosed at levels an order of
niagYnitude or more
lower in the aqueous system, yet still achieve, or build Lip to a level that
effectively
controls or removes biofilm. This lower level of biocide concentration has
positive.
effects on the environment due to the efficacy resulting from the delivery
system.
Addit:ionaal.ly~, depending upon the particular systeni that is being treated,
an embodiment
provides for flexibility iii where the liposomes are actually delivered into
the system. If
there is one particular area in a system that is prone to biofilm creation,
the delivery of
the liposomes may be delivered to that particular portion or point of the
system. such that
the delivery of the biodelivery composition is to a targeted location, and not
necessarily
privy to or exposed to the entire system. As smaller doses of the liposonie
containing
biocides are needed due to the efficacy of the biocides in this format, an
entire system of
process need :not be ..flooded with or treated with biocides.
[0027] Indeed, while the terms "antimicrobial- or "biocide" or "biocidal" have
been employed to describe the agent carried by the liposome, these agents need
not be the
highly bioactive materials normally understood by those terms, but may include
a number
of relatively harmless materials that become highly effective simply by virtue
of their
highly localized release. Thus, for example, surfactants or harmless ammonium
or
phosphonium halide salts, when released locally, may affect the normal action
of
extracellular colony-forming secretions, and are to be included as a
antiniicrobiaal or
biocidal agents for purposes of the invention, and the same mechanism may be
employed
to deliver other treatment chemicals to the targeted biofilin sites.
[0028] Aqueous systems that can be treated by this method include, but are not
limited to, potable and non-potable water distribution systems, cooling
towers, boiler
systems, showers, aquaria, sprinklers, spas, cleaning baths, air washers,
paasteudzers, air
conditioners, fluid transporting pipelines, storage tanks, ion exchange
resins, food and
beverage processing lines, metalworking fluid baths, coal and mineral
slurries, metal
leaching fluids, wastev water treatment facilities, mollusk control, pulp and
papermaking
operations, acid mine drainage, or any application prone to biofouling by
microbial
species. Application such as oil drilling,, oil storage tanks or oil
pipelines., where b:iof:ilins
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form in stagnant or t pooled aqueous sumps or lenses along the conduitsystem,
may also
be effectively treated.
[0029] Additional applications for liposome delivery of a treatment chemical
comprise natural, medical and industrial systems, such as, but not limited to
anti-
corrosion treatments for equipment generally, delivery of hormone, vitamin or
antioxidant treatments or antibiotic and gene therapies for medical or
veterinary. pur-lposes,
delivery of pesticides for agriculture:. and commercial home uses, effective
formulations
of food additives and presenvatives, targeted delivery for chemical and
biological
detection sysÃems, color and flavor enhancement, odor control, fungicides,
rodenticides,
insecticides, mildew control and aquatic pest management.
[0030] Various biocides, for example non-oxidizing biocides, can be
incorporated
into the liposome, which would be effective, The use c f certain biocides has
shown the
efficacy of this delivery system versus inclusion of biocides in the
industrial systems
wherein the biocide is outside of the liposome delivery system. The level or
concentration of biocides is measured in active levels, to provide consistency
across
various forms of the same biocide,
[001] One embodiment of the invention calls for the use of isothiazolin-3-one
biocides. These isothiazolin- 3-one liposome formulations are more effective
at killing
and removing biofzlms when compared to the same isothiazolin-3-one compounds
at the
same active concentrations, which are introduced into systems, but not
incorporated in
lipos(nnaes, as the liposonre containing biocides readily penetrate the
microbial biokilnrs
and are highlyy, eff= ctiye at destroying the bioflrmrr matrix. This lipo om.e
delivery method
may comprise 5wclrlcaro- -methyl- rise tlaircali.rti- -cane acrd - et:lrr:l- -
iscati is c~lin- -one,
but any substituted isotbiazolin-_)-one based biocide can be made
significantly more
effective when delivered in a liposome biodeliv ery system or composition.
[0032] An example of an isothiazoli.n-3-cane compound is
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lsothiazolin-3-one
Y 0
' V_~
X.. ' N -- R
Where:
R = H, Cl, Br, I. C,H'n+2)
X = H, CI, Br, 1, CnH{ ,+2
Y = H, CI, Br, 1, CnHtn+2
[0033] For an embodiment of'liposomes comprising isotlriazolin, the active
range
is from about 0.02 to about 10.0 actives, and alternately from about 0.03 to
about 5.5
active.
[0034] 1.n alternate embodiment of the invention provides for liposomes
produced that incorporate the biocide a substituted ni.trilopropiona.mide, for
example
DUNPA. The DB:NP.A liposome formulation targets and eliminates higher levels
of
biofiln when compared to the same D 3NPA compound at the sane active
concentration
that is not incorporated into liposome delivery systems. The liposome biocide
readily
penetrates the microbial biofilm and is highly effective at destroying the
biof.lm cells and
associated slime complex. This liposonre delivery method has been proven with
2,2-
dibrorno-3-nitrilo-propiona.mide, but it is believed that any substituted
nitrilcopropionaanide biocide active could be made sigill ficall tly more
effective when
delivered in a liposome format. Non-limiting examples of substituted
nitrilopropionamide
are shown. below Also, another possibility from the family of
nitrilopropionan.ide
compounds comprises :DBNlP A, 2,2,dibro..mmo-3-n-rit:rilopr-op ornarm-ride, is
also shown.
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3-Nitrilopropionamide
.1 x2 0
NC
y1 ''2 NH2
Where:
X1 =F,CI,Br,1,CH3,H
X2 = F, C1, Br, 1,CH3.H
Y., = F, Cl, Br, 1, CH3, H
Y2 = F, CI, Br, 1, CH3, H
2. -D bromo-3-nitric propionamide (DBNPA)
H B r {
NC
H Br NH2
[0035] For an embodiment of lilroscames comprising nitifloproplonamide, the
active. range is from abo. t 0,2 . to about 25 actives, and alternately from
about 0.5 to about
12.5 actives.
[0036] A further embodiment of the invention provides for liposomes that are
produced which incorporate cl. arternary ammonium salts, such as the cationic
surfactant
and biocide aikyi diÃnethyl-benzyl ammonium chloride (ADE ' [_' Qurat). :t1 BA
C type
quats are one form of ammonium salts that may be used as abiocide in the
liposonle
del:iveryr system, but any substituted quaternary ammonium salt hiocide
active, such as for
example dialkyl dimethyl clua:ts, can be more effective when delivered in a
liposome
format. 'Non-limiting examples of quaternary ammonium salts are shown with.
the
following general formula.-
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ADBAC I DIALKYL QUATS
R2
I
R1 -N-R4 X
I
R
Where Rr = 'taHi2Frr'E where n = 1 - 20
R2 + R3 = CfaHc2,+r), where n = 1 -
R4. Cfl ,2 +1), where n = 1 - 20
H
-U GH CH
Hf
CH2CH3
C1, Br, 1, I C 3, CH3OS a
[0037] For an embodiment oflipcasomes comprising anamo.nium salts, the active
range is from about 2.0 to about 2-50 actives, and alternately from about 4.0
to about 125
acfi es.
[0038] Astrrrt.her embodiment of the nveatican., comprises substituted
propanediol biocide actives, such as for example, 2-bromo,2-nitro, ],
3,propane-diol
(l3NPD active, a representative of the substituted propanediol class of
compounds.
Examples of substituted propanediol compounds:
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Substituted Propanediols
HOH2C CH2OH
y
Where:
X = CI, Br, I, NO2. O3H, OH
Y = Cl, Br, 1, N02, S03H, OH
Example:
Bronopol (BNPD) = 2--bromo- -nitre-prepanedical
= -brc e-2-nitre -l3-propanediol
Br
HOH2C +CH20H
N2
[0039] [`. ective amounts of a substituted propanediol biocide incorporated
into
the liposome would include from about 1.0 to about 100 biocide actives, or
altemately
about 2 5 to about 8.0 biocide actives.
[0040] A. f urtlaer embodiment cat` tlae :invention , l:ipr~asonres produced
drat.
incorporate the biocide phosphonium salts for example the cationic surfactant
and biocide
tributyltetradecyl plaosphonium chloride (TTPC). The TTPC' Iiposome
fibrniulation
targets and eliminates higher levels ofbiofilm when compared to the same. TTPC
compound at the same active concentration that is not incorporated into
liposome
delivery systems. The liposome biocide .readily penetrates the microbial
biofilm and is
highly effective at destroying the biofil.m cells and associated slime
corammplex. This
liposome delivery method has been proven with TT`l'K, but any phosphoniurn
salts
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biocide active could be made significantly more effective when delivered in a
liposome
format. Non-liraiting examples of phosphoniun salts are shown as:
Substituted Phosphon um Salts
R2
I
R1--P R3 X
I
Rol
Where:
RI CH3, CH9CH, CfHi2,,+1 ) where n
R2, R3. R4 = CH3, CH 0H, C,H(2,+,) where n = 2 - 5
X = Cl, Br, 1, NO2, SO4, HCI
Examples:
Bellaeide 350 (Tetradeeyl tributyl phesphonium chloride)
C~H
Cr4Hc P C4H9 Cl
CA
THPS (Tetrakis hydroxyrnethyl phosphonium sulfate)
H2OH
CN..; N-P-CH CH S0$
CH OH
[0041] Effective amounts of a plrospl>oniu salt biocide incorporated into the
liposon e would include from about 1 .0 to about 1.00 biocide acÃives, or
alternately about
1.5 to about 50.0 biocide actives
[0042] Liposor res of the present invention may be created as multi-layer
bodies.
in which one or more additional layers are provided to enhance the stability
of the
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WO 2010/107533 PCT/US2010/023973
l:iposomes or to effectuate a programmed release of the underlying lipid body
and
contents. Thus, this technology may be used to encapsulate medicines for
intracorpoaal
delivery, such that the additional layers may include a protective layer that
is hydrolyzed
or otherwise breaks down over time to provide a sustained release or longer
lifetime of
the r,rrnderlyyin. lipcosorsme. Such additional layer may additionally or
alternatively include
an encapsulating polymer that selectively breaks down when the multi-layer
liposome
encounters a low-pl--H environment, like the corrosive high acidity
environment that rnay
develop beneath a biof`rl..m. A. layer may also be compounded to be vulnerable
to sulfur
fixing bacteria, causing the liposome to specifically release its biocide in
proximity to
these corrosive organisms often present in a waste or pipeline system.
Furthermore,
several such layers may be employed to assure a sufficient lifetime of the
liposome,
prefe:raably on the order of several days as well as an ability to target a
specific niche or
environment in the biofilm. This assures that the liposomes will effectively
encounter the
target organisms or bio.film colonies and deliver their biocides thereto. The
lipid material
itself may be treated to provide enhanced resistance to hydrolysis or decay,
or the added
layers may be tuned of various harderaable or cross-linkable oils or polymers.
[0043] An alternate embodiment of the invention provides for a. biodelivery<
composition for delivering at least one antimicrobial composition into a
biotilm present
in an industrial system, wherein the biofilrn comprises at least one
microorganism
species; b) the biodelivery composition comprises a liposome structure
containing at least
one lipid or phospholipid type component, and c) the liposome structure
encapsulates at
least one antimicrobial composition.
[0044] A further embodiment provides for the targeted delivery of biocide
actives
into an industrial system, such as an industrial aqueous system, by
introducing into said
system an effective amount of said biocides in a critical area of said system,
By targeting
,in area,, and entry at a specific point in a process, the eft caacy of the
liposome system
provides for a noteworthy impact on the environment as well as the cost of
maintaining a
ay=steam, as the entire system does not need to be flooded with biocides, only
the specific
area of interest.
CA 02754820 2011-09-08
WO 2010/107533 PCT/US2010/023973
[0045] The invention will now be described with respect, to certain examples
that
are merely representative of the invention and should not be construed as
limiting thereof
EXAMPLES
[0046] The invention is illustrated in the following nonlir citing examples,
which
are provided for the purpose of representation, and are not to be construed as
limiting the
scrape of the :invention. All parts and percentages in the examples are by
weight unless
indicated otherwise.
Example 1
[4047] Three batches of liposomes (150 nanometers averagr diameter) were
created that incorporated an isothiazolin biocide, Kat-hon`r= r (available
from Rohr-
Haas, :Philadelphia, PA) as the active ingredient. The liposomes were than.
placed in
microtiter plates that had microbial biofilnis coating them. The microbe
inhibiting
efficacy of the isothiazolin liposomes was then compared with non-liposomal
ise)thiazcalina biocide when Lused at. the sarmae :isothia.zolin
conacentra.tionas. The liposonaes
containing isothiazolin penetrated the bioflrn and inhibited the hiotilm
organisms much
r tore effectively than the non--liposonial isothiazolin solution.
[0448] The .results are shown in Tables 1, 2 and 3 below and in Figs. 1, 2 and
3.
The non-liposomal isothiazolin is listed as Kathon av, each of the liposome
samples were
made by three different technicians and are referred to by code. The tables
and charts
show the. concentration of the. isothiazolin versus the percent inhibition of
the biot lm. It
is clear from both the tables and the figures that in all three trials, the
liposomal
isothiazolin formulations exhibited more effective biofilin killing)] removal
efficiency
than the isothiazolin control (listed as Kathon av) in every liposonie
concentration that
was tested. 'Elie liposome carrier is highly effective at delivering biocide
to the bioilrn at
low isothiazol.iaa. concentrations, thus providing better biofilm control at
much reduced
i sothiazolin concentrations (reduced toxicity and cost performance).
.
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Table 1
Concentration(ppm) % inhibition % inhibition % inhibition % inhibition
JIM WKW GT Kathon av
0016 21 29.9 7.9 0.93
0.031 29.7 34.1 28.7 10.9
0.0625 26.8 31 31.4 14.5
0.125 31.7 38.3 26.5 11.3
0.25 18.6 32.9 37.9 6.7
--------------- ----------------- ------
0.5 37,4 32.4 37.5 9.0
-------- --------- -------- -------- ---- ---- --------------------------------
--------- ----------------------------------------- ---------------------------
---------------
1 42.9 50.3 44.8 17.9
2 48.6
53.1 54.4 37.9
Table 2
oncentration(ppm) % inhibition % inhibition % inhibition % inhibition
JIM WKW T Kathon av
-------- -------- -- -- ----- -------- -----
0016 15.7 21.1 10.2 0.93
------------------------------------------------------------ ------------------
----------------------- ----------------------------------------- -------------
---------------------------- ------------------------------------------
0.031 27.3 31.3 26.1 10.9
0.0625 21.6 30.5 25.8 14.5
--------------------------- -------------------------------- ------------------
------------------------ ---- -
0.125 26.7 35.1 29.6 11.3
0.25 24.6 36,6 33.4 6.7
0.5 32.6 34,6 31.8 9.0
1 36.6 43.9 35.9 17.9
2 45.3 45.1 48.3 37.9
CA 02754820 2011-09-08
WO 2010/107533 PCT/US2010/023973
Table 3
Concentration(ppm) % inhibition % inhibition % inhibition % inhibition
JIM WKW GT Kathon av
0016 10.4 123 12.4 0.93
10.031 24.9 28.4 23.5 10.9
-- ----- -----------------------------------------
U625 16.3 30 221 14.5
0.125 21.E 31.9 32.7 11.3
0.25 33.5 40.2
28.8 6.7
--------------- ----------------- ------
0.5 277 36.7 26.0 9.0
-------- --------- -------- -------- ---- ----- -------------------------------
---------- ----------------------------------------- --------------------------
----------------
1 30.3 37.3 26.9 17.9
2 42.0
37,0 42.1 37.9
Example 2
[0049] One batch of liposomes (150 manometers average diameter) was created
that incorporated a substituted nitrilopropionamide ocide as the active
ingredient. The
liposo.mes were then placed in :Ã Ãacrcatiter plates that had microbial hiof
lms coating them.
The microbe inhibiting efficacy of the substituted nitrilopropfonamidc
liposomes was
then compared with non=-liposonaal substituted r itrilopr-opionanide biocide
when used at
the same n trilopropionamide concentrations. The liposoines containing
Substituted
nitrflopropionam:ide, particularly 1).BN:PA,, penetrated the b1ofilyn and
inhibited the
biofilm organisms ..much more effectively than the nova-fliposomal substituted
nitrilopropionannide solution.
[0060] The results are shown in Table 4 below and in Fig 4- The table a7.nd.
Chart
show the concentration of the substituted nitrilopropionamide versus the
percent
inhibition of the hiotil.m. It is clear from both the table and the figure
that the liposomal
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WO 2010/107533 PCT/US2010/023973
substituted .nitrilol)ral-iarrart ide f rrrrulation exhibited more effective
biofilni killing,"
removal efficiency than the substituted nitrilopropionaride control in every
liposome
concentration that was tested.
Table 4
"
""
oncentratior DBNPA liposome DBNPA
0.39 29.9 17.9
------------------------------------------------------- -----------------------
---------------------------------------- --------------------------------------
--------------------------------------
Q.B - -------- ---- 38.6 23.3
1.56 62.1 35.3
3.13 79.7
66.5
6,25 85.6 80.3
12.5 97.6 89.3
------- -------- -------- ------- -- -------- -------- ---------
Example 3
[0051] Two batches of lilaosoames (150 nanometers average diameter) were
created that incorporated an annnoniuna silt biocide as the active ingredient,
specifically
a quaternary ammonium salt, 50% allkyl,d: methyl-benzvi amnion: um chloride
(ADBAC).
The liposones were then placed in microliter plates that had microbial
biofilms coating
thenp. The microbe inhibiting efficacy of the A[).AÃ` liposomes was then
compared
with non -liposomal ADBAC biocide when used at the same BAC`: concentrations,
The liposomes containing ADBAC penetrated the biolilm and inhibited the
biofilm
organisms much more effectively than the non-liposomal ADBAC solution.
[0052] The results are shown in Tables 5 and 6 below and in Figs 5 and 6. The
table and chart show, the concentration of ADBAC'. versus the percent
inhibition of the
biofilna. It is clear from both the table and the figure that the liposonlal
ADBAC
formulations were as good as or more effective in biofilni killing! removal
efficiency than
the ADBAC control in even., liposome concentration that was tested.
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iConcentraton 50% ADBAC Quat 50%AD AC Quat
liposome
3.9 17 15
41 30
-------------------- -- - -----------------------------------------------------
----------------------
15.6 65 47
31.3 69 52
-------------------------------- 62.5 ------- --------------------------------
63-----------------------------------------
74
------------------------------------------------------- -----------------------
------------------------------------------------------ ------------------------
----------------------------------------------------
----------------------
125 91 90
95 95
250
Soo 98 97
Table 6
-------------------------------------------------------------------------------
-------------------------------------------------------------------------------
----------------------------------------------------------------------
Concentration 50% ADBAC Quat 50% ADBAC Quat
lipe one
----
3.9 7 3
7.3 15 7
15.6 40 19
-------------------------------------------------------------------------------
------------------------------------------------------- 31.3 48
21
62.5 61 40
-----------------------------
125 79 73
250 88 87
----------------------------------------------------------------------------- -
-------------------------------------------------------------------------- ----
------------------------------------------------------------------------
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[Example 4
[0053] Two batches of liposomes (1.50 nanorY eters average diameter) were
created that
incorporated a substituted propanediol biocide as the active ingredient,
specifically a
b:r-onopol. The Iiposo.mes were then placed in .microtiter plates that had
microbial bioti:laars
coating them. The microbe inhibiting efficacy of the bronopol liposonaes was
then
compared with non--liposomal bronopol biocide when used at the same bronopol
concentrations. The liposomes containing bronopol penetrated the biofllm and
inhibited
the bio.him organ isn s much more effectively than the 11011-liposomal
bronropol solution.
[0054] The results are shown in Tables 7 and 8 below and in Figs 7 ,and S. The
table and chart show the concentration of bronopol versus the percent
inhibition of the
blot lm, lt. is clear #rcart tae tla the table and tlr figure tlatat tlrc lil
c sc r rrrl larrarrc ta<al
formulations were as good as or more effective in biotilra killing/ removal
efficiency than
the br-onopol control in every l:iposomrre concentration that was tested.
Table 7
----------------------- ------- -------------------------------------------- --
-------------------------------------------------------------------------- ----
------------------------------------------------------------------------
oncentration Bronopol liposorn Bronopol
0,625 0 0
1.25 5 0
- ----------------------------
2. 19
--------------------------------------------------------------------------
24 8
---------------------------------------------------------------------------- --
-----------------------------------------------------------------------
------------------------------------------------------------------------------
35 16
48 18
------- -------- ------- -------------------------------- --- -------- -
41 53 26
-------- --------- --8 --------- -------- ------ --------- 86------------------
----------------------- ----------------------------------- 41 ----- ---------
---
22 --
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WO 2010/107533 PCT/US2010/023973
`T'able 4
Concentration Bronopol liposome ronopol
0.78 0 0
1.6 4 0
------------------------------------------------------ ------------------------
------------------------------------------------- -----------------------------
-------------------
----------------------
a 18 3
12.5 21 5
11
25 37
50 ----
44 1
100 67 32
Example 5
[Ã3065] One batch of liposomes (150 narnorneters average diameter) was created
that
incorporated a phosphonium salt biocide, Bellacide 3SOT r WA, Tucker, GA) as
the
active ingredient. The liposomes were then placed in microtiter plates that
had microbial
biolilr-rms coating them. The microbe inhibiting efficacy of the ph.osphoniur-
rm salt
liposomes was then compared with non-liposomal phosphonium salt biocide when
used
at the same concentrations. The liposomes containing phosphonium salt
penetrated the
biotilm and inhibited the biohlm orga.nisrns much r-nore effectively than the
non-
liposomal phosphonium salt solution.
[0056] The results are shown M. Table 9 below and in Fig 9. The table and
chart
show the concentration of the phosphonium salt versus the percent inhibition
of the
biotalm_ It is clear from both the table and the figure that the liposornal
phosphornium salt
formulation exhibited equal or more effective biofil m killing./ removal
efficiency than the
phosphonium salt control in every liposome concentration that was tested.
Table 9
CA 02754820 2011-09-08
WO 2010/107533 PCT/US2010/023973
Concentration Belleclde 350 Boone à Bellacide 350
039 0 0
------------------------------------------------------- ---------------
155 6.6 0
3.13 14.5 0
6,25 14.9 0
------- -------- -------- ------- -------------------------------- -----
---- ------
1 .5 23.8 8
--------- ------- --------- ---------
25 21.6
50 52 48
------- -------- -------- -------- --------- -------- -------- -------- -------
- ------- -- -------- -------- ---------
[0057] While the present invention has been described with references to
preferred embodiments, various changes or substitutions may be made on these
embodiments by those ordinarily skilled in the art pertinent to the present
invention with
out departing from the technical scope of the present invention. 'T'herefore,
the technical
scope of the present :invention encompasses not only those embodiments
described above.,
but also all that fall within the scope of the appended claims.
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