NZ753660A - Compositions and methods comprising a polyamine - Google Patents

Abstract

Compounds, compositions, and methods comprising a polyamine compound are described, which may be used to kill, disperse, treat, or reduce biofilms, or to inhibit or substantially prevent biofilm formation. In certain aspects, the present invention relates to compounds, compositions, and methods comprising polyamine compounds that have antimicrobial or dispersing activity against a variety of bacterial strains capable of forming biofilms.

Description

COMPOSITIONS AND METHODS COMPRISING A POLYAMINE CROSS-REFERENCES TO RELATED APPLICATIONS This application claims the benefit of US. Provisional Application Nos. 6l/826,453 (filed May 22, 2013), 61/826,761 (filed May 23, 2013), 61/834,149 (filed June 12, 2013); 61/836,555 (filed June 18, 2013), 61/887,267 (filed October 4, 2013), 61/902,135 (filed November 8, 2013), and 61/938,111 (filed February 10, 2014), as well as US. Non- Provisional Application Nos. 14/076,143 and 14/076,149 (both filed November 8, 2013).

These applications are incorporated by reference in their entirety for all purposes.

FIELD OF THE INVENTION

[0002] The present invention is directed to polyamine compounds, itions, and methods, which preferably have antimicrobial or dispersing ty against a variety of bacterial strains capable of forming biofilms. s aspects and embodiments relate generally to polyamine compounds and to methods of preparing or using such compounds.

BACKGROUND OF THE INVENTION

[0003] Antimicrobial compounds, such as traditional antibiotics, have the ability to kill or to retard the growth of bacteria, fungi, and other microorganisms. Some antimicrobial compounds also are effective t viruses. crobial compounds are used in a wide variety of clinical settings, industrial ations, food production facilities and environmental applications all across the globe in an effort to reduce the risk of, for example, bacterial colonization and development of e in people.

Traditional antibiotics are primarily tives or synthetic mimics of natural compounds secreted by bacteria, plants, or fungi. These nds typically have very specific s of action against a cell wall/membrane component of bacteria, or an enzyme/protein in a metabolic pathway. Examples of traditional antibiotics on the market include penicillin, oxacillin, vancomycin, gentamicin, icin and amoxicillin, among others.

Because bacteria have the ability to develop resistance genes to these antibiotics as a result of genetic mutations or acquired defense mechanisms that target the c activity of the antibiotics, bacteria lly have the ability to develop resistance to traditional antibiotics. Increasingly more prevalent bacterial resistance has made traditional antibiotics to become less and less effective in a variety of ations.

Bacterial resistance to antibiotics represents one of the most underappreciated s to modern y. See Zhang et al., Antibiotic resistance as a global threat: Evidence from China, Kuwait and the United States, Global Health 2, 6 (2006). Currently, more than 90% of clinical isolates of Staphylococcus aureus display resistance to penicillin. See Balaban et al., l ofBiofilm ions by Signal Manipulation, Ch. 1, 1-11 (Springer, 2008). Recent reports have even indicated that ia in natural ecosystems metabolize antibiotics as an energy source. See Leslie, Germs Take a Bite Out ofAntibiotics, Science 320, 33 (2008). The trend of bacterial resistance ues to increase as indicated by almost daily ific publications and world news reports of antibiotic resistant superbugs such as carbapenem-resistant bacteriacea, vancomycin-resistant Enterococci, multidrug- resistant Pseudomonas aeruginosa and methicillin—resistant Staphylococcus aureus (MRSA).

See e. g., FoxNews.com. Europe in the Grip ofDrug-Resistant Superbugs ; Melnick, M., TIME (2010); Arias et al., The rise of the Enterococcus.‘ beyond vancomycin resistance, Nat Rev Microbiol 10, 8 (2012); Jain, R. et al., Veterans aflairs initiative to prevent methicillin-resistant Staphylococcus aureus infections, N Engl J Med 364, 1419-1430 (2011); Nordmann et al., The real threat ofKlebsiella pneumoniae carbapenemase—producing bacteria, Lancet Infect Dis 9, 228-236 (2009); Aloush et al., Multidrug-resistant Pseudomonas aeruginosa.‘ riskfactors and clinical , Antimicrob Agents Chem 50, 43- 48 (2006). In addition to adversely affecting civilian patients, antibiotic-resistant ia affect injured military personnel. Multiple reports from Operation Iraqi Freedom/Operation Enduring Freedom have indicated that multidrug-resistant bacteria and otic resistance constitute one of the most disconcerting aspects of military theater treatment. See e.g., Calhoun et al., Multidrug—resistant Organisms in Military Woundsfrom Iraq and Afghanistan, Clinical Orthopaedics and Related Research 466, 1356-1362 ; Murray et al., Bacteriology of War Wounds at the Time ofInjury, ry Medicine 171, 826—829 (2006); Huj er et al., Analysis ofAntibiotic Resistance Genes in Multidrug—Resistant Acinetobacter sp. Isolatesfrom Military and Civilian Patients Treated at the Walter Reed Army Medical Center, Antmicrobial Agents and Chemotherapy 50, 4114-4123 (2006).

Multiple factors contribute to bacterial cells’ ability to resist the s of antibiotics. See e. g., Morita et al., Antibiotic Inducibility ofthe MexXYMultidrug Efflux System ofPseudomonas nosa: Involvement ofthe Antibiotic-Inducible PA54 71 Gene Product, Journal of Bacteriology 188, 1847-1855 (2006); Tran et al., Heat-Shock Protein ClpL/HSPI00 Increases Penicillin nce in ococcus pneumoniae, Advances in Oto-rhino-laryngology 72, 126-128 (2011); Livorsi et al., nce Factors ofGram- Negative Bacteria in Sepsis With a Focus on Neisseria meningitidis, Contributions to Microbiology 17, 31-47 (2011); Nostro, et al., Specific Ion Eflects on the Growth Rates of Staphylococus aureus ana1 Pseudomonas nosa, Physical Biology 2, 1-7 (2005).

Amongst these factors is the y of bacteria to develop a biofilm. See e. g., Costerton et al., How bacteria stick, Sci Am 238, 86-95 (1978); Lawrence et al., l sectioning of microbial s, J Bacteriol 173, 6558-6567 ; ZoBell, The Eflect ofSolia1 Surfaces upon Bacterial Activity, Journal of Bacteriology 46, 39-56 (1943). Biofilms have unique characteristics that allow them to withstand, or defend themselves against a variety of bations including exposure to antibiotics.

Biofilms are surface-attached ities of bacteria, often polymicrobial, that produce a slimy, extracellular polysaccharide substance (EPS) that encapsulates them. The EPS provides protection, Leid et al., The Exopolysacharide Alginate Protects Pseudomonas aeruginosa Biofilm Bacteriafrom Mediated Macrophage Killing, The Journal of Immunology 175, 7512-7518 (2005), as well as a e of nutrients, water and trace elements to sustain life. ton et al., The Bacterial Glycocalyx in Nature and Disease, Annual Review of Microbiology 35, 4 (1981). Biofilms are the inant phenotype of bacteria in natural ecosystems. egative bacteria, Gram-positive bacteria, and mycobacteria, in addition to other unicellular organisms, can produce biofilms.

Within the biofilm community, bacteria may have several methods of defending themselves against the biocidal effects of antibiotics. First, they have strength in numbers.

Biofilms may contain millions or trillions of cells in a very small volume. Second, bacteria in a biofilm have the ability to rapidly transfer genetic material, such as plasmids, that specifically code for the production of les that protect them against antibiotics. Lujan etal., Disrupting Antibiotic Resistance Propagation by Inhibiting the Conjugative DNA Relaxase, PNAS 104, 12282—12287 ; Lederberg et al., Gene Recombination in Escherichia coli. Nature 158, 529-564 (1946). Rates of plasmid transfer in biofilms have been shown to be much higher than amongst planktonic bacteria, which are free-floating in an environment. Hausner et al., High Rates ofConjugation in Bacterial Biofilms as Determined by Quantitative In Situ Analysis, Applied and Environmental Microbiology 65, 3710-3713 (1999). Third, as a m community matures, it creates an oxygen gradient such that an -rich environment exists on the outer edges of a biofilm, whereas an oxygen-deprived, or anaerobic, area exists in the deepest ns of a biofilm. Walters et al., Contributions ofAntibiotic Penetration, Oxygen Limitation, and Low Metabolic Activity to Tolerance ofPseua’omonas aeruginosa biofilms t0 Ci'profloxacin ana1 Tobramycin, Antimicrobial Agents and Chemotherapy 47, 317-323 (2003); Borriello et al., Oxygen Limitation Contributes to Antibiotic nce ofPseudomonas aeruginosa in Biofilms, Antimicrobial Agents and Chemotherapy 48, 2659-2664 (2004). This may result in reduced lic activity in those cells that dwell in the interior of the biofilm. Importantly, traditional antibiotics are typically ive against bacterial cells that are rapidly dividing, i.e., in a logarithmic phase of growth. Mandell, Interaction ofIntraleukocytic Bacteria and Antibiotics, The Journal of Clinical Investigation 52, 1673—1673 (1973); Gilbert et al., Influence ofGrowth Rate on Susceptibility to Antimicrobial Agents: Biofilms, Cell Cycle, Dormancy, ana1 Stringent Response, crobial Agents and Chemotherapy 34, 1865-1868 (1990). Fourth, in a mature m, water channels form throughout the community.

Stoodley et al., Liquidflow in biofilm systems, App Env Microbiol 60, 2711-2716 (1994).

These water channels have the ability to dlffilSC, remove or t toxic byproducts as well as antibiotics from interacting with cells in the biofilm. For novel antimicrobial agents to be effective over the long term, addressing each of these four characteristics may increase the potential for success in a variety of applications including healthcare, industrial, environmental, agricultural and sanitation industries. rmore, biofilms tend to secrete proteoglycan materials that create an extracellular matrix, which has the y to potentially bind and hinder the activity of antibiotics. ative approaches to killing ia include the use of antimicrobial agents that have fast-acting and nonspecific mode of activity against the cell membrane of bacteria.

These alternate compounds include detergents, squalamine, quaternary ammonium compounds, and lly occurring antimicrobial peptides, among others. By attacking and depolarizing the cell membrane in a nonspecific fashion at a faster rate, agents that attack the cell membrane globally can kill bacteria before they have time to upregulate their defense mechanisms. In addition, modes of action of these alternate crobials are not limited to a specific protein or enzyme within a metabolic pathway. r, as important as it is to kill bacteria and prevent their ability to cause infections in humans or animals, or inate ed processes in industrial, agricultural or environmental applications, when bacteria are attached to a surface, it sometimes may be more beneficial to not only kill bacteria, but also to cause them to “fall off’ of a surface as well, 6. g. disperse or ge bacteria in a biofilm community. In certain aspects, the present invention provides compounds, compositions, and methods that have shown the ability to disperse or dislodge bacterial cells in a biofilm, such that the cells are no longer able to reattach and form new biofilm communities, and, notably, the same compounds, compositions, and methods kill substantially all ia cells in a biofilm.

By dispersing a biofilm and killing the cells within it, at least two benefits are provided. This may be particularly important when considering the fact that although bacteria in a biofilm, which may be attached to a surface, can be killed by an antimicrobial agent, the dead cells and extracellular matrix residues may provide an attachment point for viable bacteria to re-adhere and form a biofilm once again with greater y. If biofilms are dispersed and killed, viable bacteria that are introduced to a surface will have reduced ability to preferentially adhere to that area. This can be particularly important in industrial applications wherein the formation of biofilms on a surface can be problematic, as well as medical applications wherein bacteria may adhere to the surface of a medical device. It has been surprisingly discovered that compositions of the t invention have cant potential to eradicate bacteria within a biofilm as well as cause the biofilm to se or dislodge, resulting in a y of potential applications across multiple settings.

Thus, there is a need for novel compounds, compositions, and methods that have potent antimicrobial and anti-biofilm activity against a variety of bacterial strains, especially at high bacterial concentrations and against antibiotic-resistant ia.

BRIEF SUMMARY OF THE INVENTION It is an object of the present ion to provide novel compounds, compositions, and methods having antimicrobial ty and dispersing activity t a wide y of bacterial strains capable of forming biofilms. In some preferred aspects, the invention provides compounds, compositions, and methods that are effective against antibiotic—resistant ial biofilms.

It has been discovered that nds, compositions, and s of the present invention rapidly disperse biofilms and kill microorganisms such as bacteria, so that the microorganisms do not have an opportunity to upregulate their defense mechanisms. Thus, there may be a reduced risk of bacteria developing resistance to the compounds, compositions, and s of the present invention. Furthermore, such compounds, compositions, and methods may not be limited to eradicating bacteria that are in log-phase growth. The ability of compounds, compositions, and methods of the t invention to disperse biofilms while demonstrating antimicrobial activity may address many of the characteristics that make biofilm communities difficult to treat using traditional antibiotics.

More specifically, by dispersing and killing bacteria in a biofilm, water channels and the bacterial community as a whole may be broken apart, allowing for broader distribution of antimicrobial agent(s) to a greater number, or even substantially all, of the cells within a biofilm.

Aspects of this disclosure feature methods of g, dispersing, dislodging, treating, and reducing biofilms as well as preventing or inhibiting biofilm formation. In some ments, the method comprises exposing a biofilm to an effective amount of a composition of the present invention to thereby kill, disperse, dislodge, treat, , prevent, or inhibit ial biofilms. [0016a] According to an embodiment, there is provided a non-therapeutic method for dispersing or killing a biofilm, the method comprising a step of contacting the biofilm with an anti-biofilm composition, y sing or killing the biofilm; wherein the anti-biofilm composition comprises a biocidal polyamine nd selected from the group consisting of Ra Ra A1 A5 A1 A5 A3 Ra , Ra A3 Ra , and a salt thereof; wherein: A1, A2, A3, and A5 are each an independently selected CR5; each Ra is an independently selected group of Formula V: R1a R1b N R2c R4 Rm R2d R3 ; each R1a and R1b is a member ndently selected from hydrogen and alkyl; each R2a, R2b, R2c and R2d is a member independently ed from the group consisting of hydrogen, alkyl, and fluoroalkyl; 16584576_1 (GHMatters) P41348NZ01 each R3 is a member independently selected from the group consisting of -Z1-R4, -Z1-Y1-R4, -Z1-Y1-Y2-R4, and -Z1-Y1-Y2-Y3-R4; each Y1, Y2, and Y3 is an independently selected group of Formula IA: Rm R2d each Z1 and Z2 is an independently selected -N(R4)-; each Rm is -CH2-; each m is an integer independently selected from 1 to 2; each R4 is a member ndently selected from the group consisting of hydrogen, alkyl, fluoroalkyl, alkenyl, l, aryl, cycloalkyl, heteroaryl, arylalkyl, cycloalkylalkyl, and heteroarylalkyl; or, alternatively, for an -N(R4)2 group, one of the two R4 in the group is a member selected from the group consisting of -(CO)OR6a, -(CO)N(R6a)(R6b), and -C(NR6a)N(R6b)(R6c); or, alternatively, for an -N(R4)2 group, the two R4 groups join to form a heterocyclic ring; each R5 is a member independently selected from the group consisting of hydrogen, alkyl, yl, alkoxy, aminoalkoxy, alkylamino, alkylaminoalkoxy, alkenyl, alkynyl, aryl, aryloxy, arylamino, cycloalkyl, cycloalkoxy, cycloalkylalkoxy, cycloalkylamino, lkylalkylamino, heterocyclyl, heterocycyloxy, heterocycylamino, halo, haloalkyl, fluoroalkyloxy, aryl, heteroaryloxy, heteroarylamino, arylalkyl, arylalkyloxy, arylalkylamino, heteroarylalkyl, heteroarylalkyloxy, heteroarylalkylamino, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; and each R6a, R6b, and R6c is a member independently selected from en and alkyl; wherein if R4 is -(CO)OR6a, R6a is alkyl; wherein the polyamine compound comprises at least six primary or secondary amino groups. [0016b] In another embodiment, the present invention provides an anti-biofilm composition comprising a biocidal polyamine compound used in methods according to the invention.

In an embodiment, the present invention provides a polyamine compound.

[0018] In a further embodiment, the t invention provides a composition for treatment of biofilms, the composition comprising, consisting of, or consisting essentially of a ine compound as set forth in any of the embodiments, aspects, or ation of aspects herein. 16584576_1 (GHMatters) P41348NZ01 In a r embodiment, the present invention provides a method of ng a biofilm comprising, consisting of, or consisting essentially of the step of administering a polyamine compound, or a composition comprising the polyamine compound, as set forth in any of the embodiments, aspects, or combination of aspects herein.

In a further embodiment, the present invention provides a method of making a polyamine compound, or a composition sing, consisting essentially of, or ting of the polyamine compound, as set forth in any of the embodiments, aspects, or combination of aspects herein.

These and other objects, aspects, and embodiments will become more apparent when read with the following detailed description and drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Various ments and aspects of the present invention are shown and described in reference to the numbered drawings. 16584576_1 (GHMatters) P41348NZ01 shows a simplified schematic entation of one embodiment of a class of polyamine compounds. through show exemplary starting materials or reactants that may be used to prepare certain specific embodiments ofpolyamine compounds as set forth herein. shows exemplary polyamine chains that may be used to prepare certain specific embodiments of polyamine compounds as set forth . through show exemplary polyamine compounds ing to aspects of the present invention. through show exemplary polyamine compounds according to aspects of the present ion. through show exemplary ine compounds according to aspects of the present invention. shows an ary synthesis strategy to produce polyamine compounds according to certain aspects of the present invention.

[0030] shows another exemplary synthesis strategy to produce polyamine compounds according to certain s of the present invention. shows still another exemplary synthesis gy to produce polyamine compounds according to certain aspects of the present ion. shows yet another exemplary synthesis strategy to produce polyamine compounds according to n aspects of the present invention. shows the minimum inhibitory concentration (“MIC”) of polyamines and novel polyamine compounds according to n aspects of the present ion.

A is a table showing certain polyamine compounds according to certain aspects of the present invention;

[0035] B is a table showing the effect biofilm eradication concentration (“EBEC”) of certain polyamine nds represented in and EBECs of certain polyamines. shows an example of microbial contamination in the oil and gas industry.

A shows a representative bacteria biofilm.

B shows a representative bacteria biofilm, such as the biofilm shown in A, treated according to certain standards in the oil and gas industry.

C shows a representative bacteria biofilm, such as the biofilm shown in A, treated with a ine compound of the present invention. shows the results of ng biofilms of Alcam'vorax borkumensz's grown on the surface of ized steel with a ine nd of the present invention. shows a photograph demonstrating the ability ofVanicream-based cream to elute compounds CZ-25, CZ-52, and polymyxin B polytherapy formula and to eradicate bacteria as indicated by zones of clearing around the cream. The second row of plates is from a duplicate test of the same cream. shows scanning electron microscopy (SEM) images of Ti coupons with MRSA s exposed to CZ-25 (left), CZ-52 (middle), or water only (right). The compounds CZ-25 and CZ-52 demonstrated the ability to disperse the majority of biofllms such that a monolayer of cells remained on the surface of the Ti. Those samples treated with water only had biofilm communities that remained in all areas of the coupons. shows photographs ofMRSA biofilms being dispersed by the compound CZ-25. (Left) s of MRSA grown on the surface of Ti coupons in a CDC biofllm r exposed to water for 2 hours. (Right) Biofilms ofMRSA grown on the surface of Ti coupons exposed to the nd CZ—25 in water for 2 hours. Note the strings of biofilm dispersing from the surface of the metal. shows SEM images of biofllms ofAlcanivarax barkumensz's grown on the surface of galvanized steel in a flow system, then exposed to the compound CZ-25 (also known as PBC—25). shows photographs SEM images of biofilms nivorax borkumensz's grown on the surface of galvanized steel in a flow system, then exposed to the compound CZ- 7 (also known as PBC-7). Glutaraldehyde, the standard of treatment in oil and gas facilities, was compared as a control. shows a schematic of the stir tank biofilm reactor. shows a photograph of a glass slide removed after 6 days in the stir tank bioreactor. shows a hypothetical mode of binding of spermidine to an exopolysaccharide. shows several polyamine-containing natural products with antimicrobial activity. shows a method for making several embodiments of the polyamine shows a method for making yet another embodiment of the polyamine compound. shows a method for making still another ment of the ine compound. shows a method for making several other ments of the polyamine compound. shows a pH profile of selected polyamine compounds’ hydrochloride salts in deionized water.

I5 [0055] FIGS. 30A to 301 show the effects of the polyamine compounds CZ-86 and CZ—l 10 on various bacterial cell es. B shows a control without added polyamine compounds.

It will be appreciated that the drawings are illustrative and not limiting of the scope of the invention, which is defined by the appended claims. The embodiments shown accomplish various aspects and objects of the invention; however, it will be understood that other aspects, features or modifications may be within the scope of the appended claims. It is appreciated that it is not possible to clearly show each element and aspect of the invention in a single figure, and as such, multiple figures are presented to separately illustrate s details of the invention in greater clarity. Similarly, not every embodiment need accomplish all advantages of the present invention.

DETAILED DESCRIPTION OF THE INVENTION The invention and accompanying drawings will now be sed in nce to the numerals provided n so as to enable one skilled in the art to practice the present invention. The skilled artisan will understand, however, that the inventions described below can be practiced without employing these specific details, or that they can be used for purposes other than those described . Indeed, they can be modified and can be used in conjunction with products and techniques known to those of skill in the art in light of the present disclosure. The drawings and descriptions are intended to be exemplary of various aspects of the invention and are not intended to narrow the scope of the appended claims. rmore, it will be iated that the drawings may show aspects of the invention in isolation and the elements in one figure may be used in conjunction with elements shown in other figures.

It will be appreciated that reference throughout this specification to aspects, es, advantages, or similar language does not imply that all of the aspects and advantages that may be realized with the present invention should be or are in any single embodiment of the invention. Rather, language ing to the aspects and advantages is understood to mean that a specific aspect, feature, advantage, or characteristic described in connection with an embodiment is included in at least one embodiment of the present ion. Thus, discussion of the aspects and advantages, and similar language, throughout this specification may, but does not necessarily, refer to the same embodiment.

The bed aspects, features, advantages, and characteristics of the invention may be combined in any suitable manner in one or more fiarther embodiments. Furthermore, one skilled in the relevant art will ize that the invention may be practiced t one or more of the specific aspects or advantages of a particular embodiment. In other instances, additional s, features, and advantages may be recognized and claimed in n embodiments that may not be present in all embodiments of the ion.

DEFINITIONS Unless otherwise defined, all technical and scientific terms used herein have the same g as ly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials r or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety, including U.S. Appl. Nos. 61/482,522; 61/482523; 61/591,601; 61/616,944; 61/826,453; 61/826,761; 61/836,555; 61/834,l49; 13/379,191; 14/076,143; and 14/076,149 as well as Int’l Pat. Publ. Nos. , 2012/151555, and 2013/148230. In case of conflict, the present specification, including these definitions, will control.

The terms “a,” “an,” and “the” as used herein not only includes aspects with one member, but also es aspects with more than one member. For example, an ment including “a polyamine compound and an excipient” should be understood to present certain aspects with at least a second polyamine compound, at least a second excipient, or both.

The term “about” as used herein to modify a numerical value indicates a defined range around that value. If “X” were the value, “about X” would generally te a value from 0.95X to 1.05X. Any nce to “about X” specifically indicates at least the values X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Thus, “about X” is intended to teach and provide written description support for a claim limitation of, e.g., “0.98X.” When the quantity “X” only includes whole—integer values (e.g., “X carbons”), “about X” indicates from (X-l) to (X+l). In this case, “about X” as used herein specifically indicates at least the values X, X-1, and X+1.

When “about” is applied to the beginning of a numerical range, it applies to both ends of the range. Thus, “from about 5 to 20%” is equivalent to “from about 5% to about %.” When “about” is applied to the first value of a set of values, it applies to all values in that set. Thus, “about 7, 9, or 11%” is equivalent to “about 7%, about 9%, or about 11%.”

[0064] The term “acyl” as used herein includes an alkanoyl, aroyl, heterocycloyl, or heteroaroyl group as defined . Examples of acyl groups include, but are not limited to, acetyl, benzoyl, and nicotinoyl.

The term “alkanoyl” as used herein es an alkyl-C(O)- group wherein the alkyl group is as defined herein. Examples of alkanoyl groups include, but are not limited to, acetyl and propanoyl.

The term “agent” as used herein includes a compound or mixture of compounds that, when added to a composition, tend to e a particular effect on the composition’s properties. For example, a composition sing a thickening agent is likely to be more s than an otherwise identical comparative composition that lacks the ning agent.

[0067] The term “alkenyl” as used herein includes a straight or branched chain hydrocarbon ning at least one carbon-carbon double bond. The chain may contain an indicated number of carbon atoms. For example, “Cl-Cu alkenyl” indicates that the group may have from 1 to 12 (inclusive) carbon atoms and at least one carbon-carbon double bond.

When the indicated number of carbon atoms is 1, then the Ci alkenyl is double bonded to a carbon (i.e., a carbon lent to an oxo group). In certain aspects, the chain includes 1 to 12, about 2 to 15, about 2 to 12, about 2 to 8, or about 2 to 6 carbon atoms. Examples of an alkenyl group may include, but are not limited to, ethenyl (z'.e., vinyl), allyl, propenyl, butenyl, crotyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, dodecenyl, cyclopentenyl, cyclohexenyl, 2-isopentenyl, l, butadienyl, pentadienyl, 3-(l,4- pentadienyl), and hexadienyl.

[0068] An alkenyl group can be unsubstituted or optionally substituted. When optionally substituted, one or more hydrogen atoms of the alkenyl group (e.g., from 1 to 4, from 1 to 2, or I) may be replaced with a moiety independently selected from fluoro, y, , amino, alkylamino, acylamino, thio, and hio, with the proviso that no hydrogen atom substituent on the carbon-carbon double bond is replaced by a hydroxy, amino, or thio group.

In some aspects, the alkenyl group is unsubstituted or not optionally tuted.

The term “alkyl” as used herein includes an aliphatic hydrocarbon chain that may be straight chain or branched. The chain may contain an indicated number of carbon atoms: For example, C1-C12 indicates that the group may have from 1 to 12 (inclusive) carbon atoms in it. If not otherwise indicated, an alkyl group contains from 1 to about 20 carbon atoms. In some aspects, alkyl groups have 1 to about 12 carbon atoms in the chain. In some aspects, alkyl groups (“lower ) have 1 to about 6 carbon atoms in the chain. Examples may e, but are not limited to, methyl, ethyl, propyl, isopropyl (iPr), l-butyl, 2—butyl, isobutyl (iBu), tert-butyl, pentyl, 2-methylbutyl, l,l-dimethylpropyl, hexyl, heptyl, octyl, nonyl, decyl, docecyl, cyclopentyl, or cyclohexyl.

[0070] An alkyl group can be unsubstituted or optionally substituted. When ally substituted, one or more hydrogen atoms of the alkyl group (e.g., from 1 to 4, from 1 to 2, or 1) may be replaced with a moiety independently selected from fluoro, hydroxy, alkoxy, amino, alkylamino, acylamino, thio, and alkylthio. In some aspects, the alkyl group is unsubstituted or not optionally substituted.

[0071] The term “alkoxy” as used herein es a ht or branched chain saturated or rated hydrocarbon containing at least one oxygen atom in an ether group (e.g., EtO-).

The chain may contain an indicated number of carbon atoms. For e, “Cl-Cu alkoxy” indicates that the group may have from 1 to 12 (inclusive) carbon atoms and at least one oxygen atom. Examples of a C1-C12 alkoxy group include, but are not limited to, methoxy, ethoxy, isopropoxy, , n-pentoxy, toxy, neopentoxy, and hexoxy.

An alkoxy group can be unsubstituted or optionally substituted. When optionally substituted, one or more en atoms of the alkoxy group (e.g., from 1 to 4, from 1 to 2, or 1) may be ed with a moiety independently selected from fluoro, hydroxy, , amino, alkylamino, ino, thio, and alkylthio, with the proviso that no hydrogen atom alpha to the ether oxygen is replaced by a hydroxy, amino, or thio group. In some aspects, the alkoxy group is unsubstituted or not optionally substituted.

[0073] The term “alkynyl” as used herein includes a straight, branched, or cyclic hydrocarbon containing at least one carbon—carbon triple bond. Examples may include, but are not limited to, ethynyl, propargyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, or decynyl.

An alkynyl group can be unsubstituted or optionally tuted. When optionally substituted, one or more hydrogen atoms of the alkynyl group (e.g., from 1 to 4, from 1 to 2, or 1) may be replaced with a moiety independently selected from fluoro, hydroxy, alkoxy, amino, alkylamino, acylamino, thio, and alkylthio, with the proviso that no sp-hybridized hydrogen atom substituent is replaced by a hydroxy, amino, or thio group. In some aspects, the l group is unsubstituted or not optionally substituted. 2O [0075] The term “aroyl” as used herein includes an aryl-CO- group wherein aryl is as defined herein. Examples include, but are not limited to, l, naphth—1—oyl and naphth- 2-oyl.

The term “aryl” as used herein includes cyclic aromatic carbon ring systems containing from 6 to 18 carbons. Examples of an aryl group include, but are not limited to, , naphthyl, anthracenyl, tetracenyl, biphenyl and phenanthrenyl.

An aryl group can be unsubstituted or optionally tuted. When optionally substituted, one or more hydrogen atoms of the aryl group (e.g., from 1 to 5, from 1 to 2, or 1) may be replaced with a moiety independently selected from alkyl, cyano, acyl, halo, hydroxy, alkoxy, amino, mino, acylamino, thio, and alkylthio. In some aspects, the alkoxy group is unsubstituted or not optionally substituted.

The term lkyl” or “aralkyl” as used herein includes an alkyl group as defined herein where at least one hydrogen substituent has been replaced with an aryl group as defined herein. Examples include, but are not limited to, , 1-phenylethyl, 4- methylbenzyl, and 1,1 ,-dimethylphenylmethyl.

A arylalkyl or aralkyl group can be unsubstituted or optionally substituted as per its component . For example, but without limitation, the aryl group of an arylalkyl group can be substituted, such as in 4-methylbenzyl. . In some aspects, the group is unsubstituted or not optionally substituted, especially if including a defined substituent, such as a yalkyl or alkylaminoalkoxy group.

[0080] The term “cycloalkyl” as used herein includes a cyclic hydrocarbon group that may contain an indicated number of carbon atoms: For example, C3-C12 indicates that the group may have from 3 to 12 sive) carbon atoms in it. If not otherwise indicated, a cycloalkyl group includes about 3 to about 20 carbon atoms. In some aspects, cyclo alkyl groups have 3 to about 12 carbon atoms in the group. In some aspects, cycloalkyl groups have 3 to about 7 carbon atoms in the group. Examples may include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 4,4—dimethylcyclohexyl, and cycloheptyl.

A cycloalkyl group can be unsubstituted or optionally substituted. When optionally substituted, one or more hydrogen atoms of the cycloalkyl group (e.g., from 1 to 4, from 1 to 2, or 1) may be replaced with a moiety independently selected from fiuoro, hydroxy, alkoxy, 2O amino, alkylamino, acylamino, thio, and alkylthio. In some aspects, a substituted lkyl group can incorporate an exo- or endocyclic alkene (e.g., cyclohexenyl). In some aspects, a cycloalkyl group is unsubstituted or not optionally substituted.

The terms “disorder,” “disease,” and “condition” are used herein interchangeably for a condition in a subject. A disorder is a bance or derangement that affects the normal function of the body of a subject. A disease is a pathological ion of an organ, a body part, or a system resulting from various causes, such as infection, genetic defect, or environmental stress that is terized by an identifiable group of symptoms. A disorder or disease can refer to a biofilm—related disorder or disorder caused by a planktonic bacterial phenotype that is characterized by a e-related growth of bacteria.

[0083] The term “effective amount” or “effective dose” as used herein includes an amount sufficient to achieve the d result and accordingly will depend on the ingredient and its desired result. Nonetheless, once the desired effect is fied, determining the effective amount is within the skill of a person d in the art.

As used herein, “fluoroalkyl” includes an alkyl group wherein the alkyl group includes one or more fluoro- substituents. Examples include, but are not limited to, trifluoromethyl.

As used herein, “geminal” substitution es two or more substituents that are ly attached to the same atom. An example is 3,3-dimethyl substitution on a cyclohexyl or spirocyclohexyl ring.

As used herein, “halo” or “halogen” includes fluoro, chloro, bromo, or iodo.

[0087] The term “heteroaryl” includes mono and bicyclic ic groups of about 4 to about 14 ring atoms (e. g., 4 to 10 or 5 to 10 atoms) containing at least one heteroatom.

Heteroatom as used in the term heteroaryl refers to oxygen, sulfur and nitrogen. A nitrogen atom of a heteroaryl is optionally oxidized to the ponding N—oxide. Examples include, but are not limited to, pyrazinyl, furanyl, thienyl, pyridyl, pyrimidinyl, olyl, isothiazolyl, oxazolyl, thiazolyl, lyl, furazanyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,4- thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl, imidazo[l,2-a]pyridine, imidazo[2,l-b]thiazolyl, benzofurazanyl, indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl, imidazolyl, thienopyridyl, quinazolinyl, pyrimidyl, pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl, l,2,4-triazinyl, and hiazolyl.

[0088] A heteroaryl group can be unsubstituted or optionally substituted. When optionally tuted, one or more hydrogen atoms of the heteroaryl group (e.g., from 1 to 5, from 1 to 2, or 1) may be replaced with a moiety independently selected from alkyl, cyano, acyl, halo, hydroxy, alkoxy, amino, alkylamino, acylamino, thio, and alkylthio. In some aspects, the aryl group is unsubstituted or not optionally substituted.

[0089] The term “heteroaroyl” as used herein includes a heteroaryl-C(O)- group wherein heteroaryl is as defined herein. Heteroaroyl groups include, but are not limited to, thiophenoyl, nicotinoyl, pyrrolylcarbonyl, and pyridinoyl.

The term ocycloyl” as used herein includes a heterocyclyl-C(O)- group wherein heterocyclyl is as defined herein. Examples include, but are not limited to, N—methyl prolinoyl and tetrahydrofuranoyl.

As used herein, “heterocyclyl” includes a non-aromatic saturated monocyclic or multicyclic ring system of about 3 to about 10 ring atoms (e.g., 5 to about 10 ring atoms, or 3 to about 6 ring atoms), in which one or more of the atoms in the ring system is an element or elements other than carbon, e. g., nitrogen, oxygen or sulfur. A cyclyl group optionally comprises at least one spZ-hybridized atom (e.g. , a ring incorporating an carbonyl, endocyclic olefin, or exocyclic olefin). In some embodiments, a nitrogen or sulfur atom of the heterocyclyl is optionally ed to the corresponding N-oxide, S-oxide or S,S-dioxide.

Examples of monocycylic heterocyclyl rings include, but are not limited to, piperidyl, idinyl, piperazinyl, morpholinyl, rpholinyl, thiazolidinyl, l,3-dioxolanyl, 1,4- dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, and tetrahydrothiopyranyl.

A heterocycyl group can be unsubstituted or optionally substituted. When optionally substituted, one or more hydrogen atoms of the group (e.g., from 1 to 4, from 1 to 2, or 1) may be replaced with a moiety independently selected from fluoro, hydroxy, alkoxy, amino, alkylamino, acylamino, thio, and alkylthio. In some aspects, a substituted heterocycyl group can orate an exo— or endocyclic alkene (e.g., cyclohex—2—en—l—yl). In some aspects, the cycyl group is unsubstituted or not optionally substituted.

The term “hydrophobic moiety” or “hydrophobic group” as used herein includes a moiety or a functional group that repels water. Examples may e, but are not limited to, a non—polar alkyl moiety, such as an unsubstituted alkyl group having more than five carbons; a phenyl group; and an anthracenyl group.

As used herein, the terms “hydrophilic moiety" or “hydrophilic group” includes a moiety or a functional group that has a strong affinity to water. Examples may e, but are not limited to, a charged moiety, such as a cationic moiety or an anionic moiety, or a polar uncharged moiety, such as an alkoxy group or an amine group.

[0095] As used , the term “hydroxyalkyl” includes an alkyl group where at least one hydrogen substituent has been replaced with an alcohol (-OH) group. In n aspects, the yalkyl group has one l group. In certain s, the hydroxyalkyl group has one or two alcohol groups, each on a different carbon atom. In certain aspects, the hydroxyalkyl group has 1, 2, 3, 4, 5, or 6 alcohol groups. Examples may include, but are not limited to, hydroxymethyl, 2—hydroxyethyl, and l—hydroxyethyl.

When any two substituent groups or any two instances of the same substituent group are “independently selected” from a list of alternatives, the groups may be the same or different. For example, if Ra and Rb are independently selected from alkyl, fluoro, amino, and hydroxyalkyl, then a molecule with two Ra groups and two Rb groups could have all groups be an alkyl group (e.g., four different alkyl groups). Alternatively, the first Ra could be alkyl, the second Ra could be fluoro, the first Rb could be hydroxyalkyl, and the second Rb could be amino (or any other substituents taken from the group). Alternatively, both Ra and the first Rb could be fiuoro, while the second Rb could be alkyl (i.e., some pairs of substituent groups may be the same, while other pairs may be different).

As used herein, “polyamine” includes a compound that has at least two amine , which may be the same or ent. The amine group may be a primary amine, a secondary amine, a tertiary amine, or quaternary ammonium salt. Examples may include, but are not d to, 1,3-diaminopropane, 1,4-diaminobutane, hexamethylenediamine, dodecan- 1,12-diamine, spermine, spermidine, norspermine, and norspermidine.

As used herein, “or” should in general be construed non—exclusively. For example, an embodiment of “a composition comprising A or B” would typically t an aspect with a composition comprising both A and B, and an ment of “a method to disperse or kill biofilms” could disperse, kill, or a combination of both. “Or” should, however, be construed to exclude those aspects presented that cannot be ed without contradiction (e.g., a composition pH that is between 9 and 10 or between 7 and 8).

As used herein, “spirocycloalkyl” includes a cycloalkyl in which geminal substituents on a carbon atom are replaced to join in forming a l,l-substituted ring. For example, but without limitation, for a —C(R1)(R2)- group that was part of a longer carbon chain, if R1 and R2 joined to form a cyclopropyl ring orating the carbon to which R1 and R2 were bonded, this would be a spirocycloalkyl group (i.e., spirocyclopropyl).

As used herein, “spiroheterocyclyl” includes a heterocycloalkyl in which l substituents on a carbon atom are replaced to join in forming a l,l-substituted ring. For example, but without limitation, for a —C(R1)(R2)- group that was part of a longer carbon chain, if R1 and R2 joined to form a pyrrolidine ring incorporating the carbon to which R1 and R2 were bonded, this would be a spiroheterocyclyl group.

As used herein, the term “treat,” “treating,” or “treatment” includes administering or ng a composition (e.g., a composition described herein) in an amount, manner (e.g., schedule of administration), and mode (e.g., route of administration) that is effective to e a disorder or a symptom thereof, or to prevent, to retard, or to slow the progression of a disorder or a symptom thereof. Such improvements can include, but are not d to, alleviation or amelioration of one or more ms or conditions, diminishment of the extent of a disease, stabilizing (i.e., not worsening) the state of disease, prevention of a disease’s transmission or spread, delaying or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable.

This can be evidenced by, e.g., an improvement in a ter associated with a m or with a m-related disorder or an indication or symptom thereof, a biofilm- related industrial, agricultural, environmental, etc. condition, e.g., to a tically significant degree or to a degree detectable to one d in the art. An effective amount, manner, or mode can vary depending on the surface, application, or subject and may be ed to the e, application, or subject. By eradicating a biofilm or preventing or slowing progression of a biofilm or of a biofilm-related disorder or an indication or symptom thereof, or a biofilm-related industrial, agricultural, environmental, etc. condition, a treatment can prevent or slow deterioration or corrosion resulting from a biofilm or from a biofilm-related disorder or an indication or symptom thereof on an affected surface or in an affected or sed subject. ing” and “treatment” as used herein also include prophylactic treatment. In certain embodiments, treatment methods comprise administering to a subject a therapeutically effective amount of a ition of the invention. The administering step may consist of a single administration or may comprise a series of administrations. The length of the treatment period depends on a variety of factors, such as the severity of the condition, the age of the patient, the concentration of active agent in the composition, the ty of the compositions used in the treatment, or a combination thereof. It will also be appreciated that the effective dosage of an agent used for the ent or laxis may increase or decrease over the course of a particular treatment or prophylaxis . Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some aspects, chronic administration may be required. For example, the compositions are administered to the subject in an amount, and for a duration, sufficient to treat the patient.

[0104] As used herein, a reference to a composition of formula A, B, C, or a salt thereof may indicate A, a salt ofA, B, a salt ofB, C, or a salt of C.

POLYAMINE COMPOUNDS Aliphatic polyamines, for example, spermine, spermidine, norspermine, norspermidine, hexamethylenediamine, 1,12-diaminododecane, 1,3-diaminopropane, etc., have been shown to have some activity against a y of bacterial strains when used alone.

In particular, it has been shown that norspermine and norspermidine may be effective in dispersing various biofilm strains. However, it does not appear that these polyamines have sufficient bactericidal activity.

It has been surprisingly discovered that by attaching one or more polyamine chains to a lipophilic or hydrophobic moiety, examples of which may include cyclic or aromatic IO backbone molecules, results in novel compounds that are capable of dispersing biofilms and that also have substantial antimicrobial activity against a variety of s of bacteria (e. g., by killing the bacteria).

Referring now to there is shown a simplified schematic of an exemplary polyamine compound of the t ion, generally indicated at 10. The polyamine compound 10 may be generally characterized as including a lipophilic or hydrophobic moiety , and one or more ic residues 30. In some embodiments, the polyamine compound may e one or more cationic residues 30 comprising primary amines 40 and one or more secondary amines 50. In some embodiments, the polyamine compound 10 may include one or more cationic residues 30 having a tertiary or quaternary amine (not shown).

[0108] As depicted in exemplary polyamine nds 10 of the present invention may be amphipathic, e.g. the cationic residues 30 may be generally disposed along one face ofthe molecule, while the lipophilic or hydrophobic moiety 20 is generally ed along the opposite face of the molecule.

In n embodiments, the present invention provides polyamine compounds and compositions and methods comprising such compounds. In certain embodiments, the polyamine compounds comprise one or more polyamine side chains (e.g., l, 2, 3, 4, 5, 6, 7, 8, ... n polyamine moieties/side chains) that may be the same or different. In n embodiments, the amino groups of the one or more polyamine side chains may be ionizable.

In n ments, the amino groups may be positively d. In some embodiments, the ine side chains are branched. In some embodiments, the polyamine chains are linear.

FIGs. 2A-2H show ary backbone les, which may be used to prepare certain novel polyamine compounds. It will be appreciated by those skilled in the art that alternative backbone molecules may be used to produce ine compounds according to certain aspects of the present invention. Without being bound by , it is believed that the lipophilicity or hydrophobicity of a ne molecule of novel polyamine compounds may contribute to the antimicrobial activity or biofilm dispersing activity of these novel compounds. shows exemplary polyamine chains, which may be used to prepare certain novel ine compounds according to certain s of the present invention. It will be appreciated by those skilled in the art that polyamine chains are intended to only be representative. Without being bound by theory, it is believed that increasing the number of primary or secondary amines of polyamine compounds of the present invention may increase the antimicrobial activity or biofilm dispersing activity of these novel compounds.

FIGs. 4A-4P show exemplary polyamine nds sing a benzene backbones. FIGs. 5A and 5B show exemplary polyamine compounds comprising an anthracene backbone. FIGs. 6A—6E show exemplary polyamine compounds comprising a Vitamin D backbone. through show exemplary chemical synthesis strategies for preparing a variety of polyamine compounds according to certain aspects of the present invention.

[0114] In n embodiments, the one or more polyamine moieties may have at least three amino groups separated by three atoms either in a straight chain or cyclic molecule. In some ular embodiments, the one or more polyamine moiety or moieties may comprise norspermidine (also known as N—(3-aminopropyl)propane-l,3-diamine), norspermine (N'-[3- (3 -aminopropylamino)propyl]propane-l,3—diamine), or a combination thereof.

[0115] Exemplary compositions of the present invention may include a compound comprising a single polyamine chain or multiple polyamine chains attached to a base backbone compound to improve their antimicrobial activities or improve certain other features or aspects not present when polyamines are used alone such that the itions may be ed for use in a specific application. Furthermore, it has been surprisingly discovered that by increasing the number ofprimary and ary amines within a single molecule an increase in the antimicrobial activity of a compound may result. rmore, exemplary novel polyamines may be used in combination (6.g. formulated as a composition) with already approved, commercial antimicrobial products (6.g. chlorhexidine gluconate) to improve the dispersing or killing ability of such compositions.

In some aspects, the invention provides a compound or composition that comprises, consists essentially of, or consists of a polyamine compound or ition used in any of the embodiments or aspects of the methods described herein.

In some aspects, the invention provides a composition for ng (e.g., dispersing or killing) biofilrns, the ition comprising a ine compound selected from: p11J§A4 l2 l3 Ra Ra Ra Y WW gYAAB: 1&A45A: UYAiAa l2 l4 A2 A's l7 l2 i l l7 A's l7 \AB‘ \A3’ \A6’ \A3 L1 AB’ \AB' , , , ,andasalt thereof; wherein: each Ra is a member independently ed from and ; A1, A2, A3, A4, A5 , A6, A7, A8, and A9 are each an A11 member independently selected from N, CR3, and CR5; or, alternatively, a pair of adjacent AI1 members join to form an independently selected aryl, cycloalkyl, heterocyclyl, or heterocycloaryl ring that is fused with an AI1 ring at the pair’s AI1 ring positions; wherein at least one A11 member and at most five AI1 members are an independently selected CR3; each R”, Rlb, R”, and R1d is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl; or, alternatively, an R121 and an R1b join to form an oxo group; each Rza, R213, R2°, de, R23, and R2f is a member independently ed from hydrogen, alkyl, fluoroalkyl, alkenyl, l, aryl, heteroaryl, kyl, and heteroarylalkyl; alternatively, a pair of R2 members from the same Ra group independently selected from the group R2&1 and RZb, R20 and R“, and R23 and szjoin to form a member independently selected from spirocycloalkyl, spiroheterocycyl, and oxo; or, alternatively, an R2a and an ch from the same Ra group join to form a ring ndently selected from cycloalkyl and heterocycyl; each Rm is a member ndently selected from —CR23R2b-, -CR2°R2d-, -C(R2a)=(R2b)-, -CC-, and -C(Rza)(RZb)-L2-C(R2°)(R2d)-; each m is an integer ndently selected from 1 to 20; each L1 and L2 is a member independently selected from a bond, -O-, -C(O)O-, -NR4-, -NR4C(O)-, and —C(O)NR4-; each R3 is a member independently selected from , -R4,—Z1-Yl-Y2-R4, and -Z1-Y1-Y2-Y3-R4; each R4 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl, or, alternatively, for a - N(R4)2 group, one of the two R4 in the group is a member selected from -(CO)OR63-, R6a)(R6b), and 7C(NR6a)N(R6b)(R6°); or, alternatively, for an —N(R4)2 group, the two R4 groups join to form a heterocyclic ring; each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, aminoalkoxy, alkylamino, alkylaminoalkoxy, alkenyl, alkynyl, aryl, y, arylamino, cycloalkyl, cycloalkoxy, lkylalkoxy, lkylamino, lkylalkylamino, heterocyclyl, heterocycyloxy, heterocycylamino, halo, haloalkyl, fluoroalkyloxy, heteroaryl, heteroaryloxy, heteroarylamino, arylalkyl, arylalkyloxy, arylalkylamino, heteroarylalkyl, heteroarylalkyloxy, heteroarylalkylamino, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; each Y1, Y2, and Y3 is an independently selected group of Formula IA: each Z1 and Z2 is a member independently selected from NR4 and O; and each R6a, R6b, and R60 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, aryl, arylalkyl, cycloalkyl, and heteroarylalkyl, or, alternatively, two R6H s R681 and R61) or R681 and R60 join to form a heterocycyl ring; wherein the polyamine compound comprises at least two primary or secondary amino groups.

In certain aspects, each R4 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl, or, alternatively, for a -N(R4)2 group, one of the two R4 in the group is a member selected from -(CO)OR6a-, (CO)N(R63)(R6b), and —C(NR63)N(R6b)(R6°); and each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, alkylamino, alkenyl, alkynyl, aryl, aryloxy, arylamino, cycloalkyl, cycloalkoxy, cycloalkylamino, heterocyclyl, heterocycyloxy, heterocycylamino, halo, kyl, alkyloxy, heteroaryl, heteroaryloxy, arylamino, arylalkyl, arylalkyloxy, arylalkylamino, heteroarylalkyl, heteroarylalkyloxy, heteroarylalkylamino, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl.

In another preferred aspect, the compounds or compositions of the embodiments and aspects described herein has the o that the polyamine compound is not of Formula IB: R\N/\/\N4 N/\/\N/R4 F? F'z4 IB wherein the —N(R4)2 groups are tertiary amines.

In some aspects, the nds or itions of the embodiments and aspects described herein has the proviso that the polyamine compound is not: R\N/\/\N4 NMN’R4 F'z4 F'w IB-2 wherein the —R4 groups are hydrogen or methyl.

In a preferred aspect, the invention presents the compounds or compositions described herein with the proviso that the polyamine nd is not of formula IB: R\WNMN4/\/\N/R4 4 or a salt thereof; IB wherein the —N(R4)2 groups are tertiary amines; and with the proviso that the ine compound is not of formula IC: RZ-HN-(CH2)p-NH-(CH2)q-NHCH2-Y-CHZNH-(CH2)S-NH-(CH2)t-NH-RZ (IC) or a salt f; wherein Y is selected from anthracenyl, naphthyl, and phenyl; wherein each RZ is independently selected from en and alkyl; and wherein p, q, t, and s are each an integer independently selected from 3 and 4.

In some aspects, the composition comprises, consists essentially of, or ts of a polyamine compound of formula Ra Ra Ra Ra RaU 0Ra Ra Ra U L1 Ra L1 R5 Ll Ra L1 R5 R5 Ra, Ra R5 or a salt thereof; wherein: each Ra is independently a group of Formula I: each R121 and R1b is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl; each Rza, R21), R2°, and R2d is a member independently selected from hydrogen, alkyl, fluoroalkyl, aryl, and arylalkyl; each Rm is an independently selected—CRzaRZb-g each m is an integer independently selected from 1 to 2; each L1 is a member independently selected from a bond and each R3 is an independently selected -Zl-Y1-R4; each R4 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, and alkynyl; or, alternatively, for an -N(R4)2 group, one of the two R4 in the group is a member selected from -(CO)OR6a-, -(CO)N(R6a)(R6b), and -CCNR6a)N(R6b)(R6°); each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, halo, lkyl, fluoroalkyloxy,and hydroxyalkyl; each Y1 is an independently selected group of Formula IA: each Z1 and Z2 is an independently selected NR4; and each R6a, R61), and R60 is a member independently selected from hydrogen and alkyl; wherein if R4 is -C(O)OR6a, R6&1 is alkyl; and wherein the polyamine compound comprises at least two primary or secondary amino groups.

In some aspects, each m is 1. In some aspects, at least one m is 1. In some aspects, each m is 2. In some aspects, at least one m is 2.

[0124] In some aspects, each Rza, RZb, R2°, and R2d is a member ndently selected from hydrogen, alkyl, and fluoroalkyl. In some aspects, each Rza, RZb, ch, and R2d is a member independently selected from hydrogen, alkyl, fluoroalkyl, and arylalkyl.

In some aspects, each R121 and R1b is a member independently selected from hydrogen and alkyl.

[0126] In some aspects, the ine compound comprises at least four y or secondary amino .

In some aspects, the polyamine compound is of formula or a salt thereof.

In some aspects, Ra is -CH2[NH(CH2)3]2NH2. In some aspects, R3 is an independently selected group of Formula VII: H’ 7ng VII.

[0129] In some aspects, each m is l.

In some aspects, R5 is hydrogen.

In some aspects, each R4 is a member independently selected from hydrogen and alkyl.

In some aspects, the ine compound is selected from H2N[(H2C)3HN]2H2C CHleH(CH2)3]2NH2 Ph H2N[(H2(3)3|‘|N]2|‘|2C CH2)3]2NH2 and a salt thereof.

In some aspects, L1 is a bond.

In some aspects, Rm is -CH2-.

[0135] In some aspects, the polyamine compound is selected from H2N[(H20)3HN]2H2C CH2)3]2NH2 H2N[(H2(3)3|'|N]2l’|2C CHleH(CH2)3]2NH2 ll(l‘lzc)3l‘lN12H2C CHleH(CH2)3]2NH2 CHleH(CH2)3]2NH2, H2N[(H20)3HN]2H20MMCH2[NH(CH2)3]2NH2 CHleH(CH2)3]2NH2 l‘l2l\ll(l‘l20)3l‘lN12H2CMCHleH(CH2)3]2NH2 CHleH(CH2)3]2NH2 OiPr H2N[(HZC)3HN]2HZC OD CHleH(CH2)3]2NH2 H2N[(H2C3)3HN]2H2C CHleH(CH2)a]2NH2, and a salt thereof.

In some aspects, each Ra is independently a group of Formula II: R13Nlb R.2 N R20 m R3 each of the An members is independently selected from CR3 and CR5; or, alternatively, a pair of adjacent AI1 members join to form a cycloalkyl, aryl, cyclyl, or heterocycloaryl ring; wherein at least one A11 member and at most three A11 members are independently selected CR3, each R”, Rlb, R”, and R1d is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl; for each Ra member, at most two R”1 of the Ra member are selected from - C(R2a)=(R2b)-, -cc-, and -C(Rza)(R2b)-L2-C(R2c)(R2d)-; and each m is an integer independently selected from 1 to 16.

In some aspects, the polyamine compound is selected from: Ra R3 fil J§A5 fi1J§A4 A2 l4 l2 A fiS’A\\AB9iv \A3. \A3 L1 AG' and a salt thereof; wherein: at least one AIl member and at most three A11 members are ndently selected CRa, each Rza, R21), R”, R”, R23, and R2f is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl; alternatively, a pair of R211 members from the Ra group independently selected from the group R2a and R21), R20 and R”, and R2e and R2f join to form a ring ndently selected from spirocycloalkyl and eterocycyl; or, atively, the R221 and the R20 from the R3 group join to form a ring independently selected from cycloalkyl and heterocycyl; each m is an integer independently selected from 1 to 12; each R3 is a member independently selected from -Z1-Y1-R4 and -Zl-Y1-Y2-R4; and each Z1 and Z2 is an independently selected NR4.

[0138] In some aspects, A1, A2, A3, A4, A5, A6, A7, A8, and A9 are each an AI1 member ndently selected from CRa, and CR5; or, alternatively, a pair of adjacent AIl members join to form a lkyl, aryl, heterocyclyl, or heterocycloaryl ring; each Rza, R2b, R2°, RM, R26, and R2f is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl; each m is an integer independently selected from 1 to 10; each L1 and L2 is a member independently ed from a bond, -O-, and -NR4-; each R3 is a member ndently selected from -Z1-R4, -Z1-Y1-R4, and -Z1-Y1-Y2-R4; and each Z1 and Z2 is an independently selected NR4.

[0139] In some aspects, the invention provides the polyamine compound selected from: Ra R8 fi1/KT5 P11J§A4 5’A§Ae9 ' ' A\ 3—.A4 ‘l 2 A2 ,1 —.A7 A \A3 L1 A6 and a salt thereof; , , wherein: each R"1 is independently a group of Formula V: N R20 R“: ( Rm%R2d m R3 each of the An members is independently selected from CRa and CR5; or, alternatively, a pair of adjacent AIl members join to form a cycloalkyl, aryl, heterocyclyl, or heterocycloaryl ring; wherein at least one A11 member and at most three A11 members are ndently selected CRa, each Rla, Rlb, R”, and R1d is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl; for each Ra member, at most two R”1 of the Ra member are selected from — C(Rza)=(R2b)-, —cc—, and -C(Rza)(R2b)-L2-C(R2°)(R2d)-; and each R3 is a member independently selected from -Zl-Y1-R4 and -Zl-Y1-Y2-R4.

[0140] In some aspects, the polyamine compound ses at least four primary or secondary amino groups. In some aspects, the polyamine compound comprises at least six primary or secondary amino groups.

In some aspects, the ion provides the polyamine compound selected from Ra Ra 1km 1kg) Elam/k 13,1 Ra, Ra A Ra, and a salt f; wherein each A11 member is an ndently selected CR5.

In some s, the invention provides the polyamine compound selected from Ra Ra R5 and a salt thereof; and wherein R3 is an independently selected group of Formula VII: HI ( Rm)'\ m R3 VI].

In some aspects, the invention provides the polyamine compound selected from Ra R3 R5 and a salt thereof; and wherein R5 is hydroxyl, alkoxy, cycloalkoxy, or arylalkyloxy.

In some aspects, Rm is —CH2-. In some aspects, Ra is —CH2[NH(CH2)n]pNH2; each n is an integer independently selected from 3 to 12 ; and each p is an integer ndently selected from 1 to 3. In some aspects, m is 1. In some aspects, each m is 1. In some aspects, at least one In is 1. In some aspects, each m is 2. In some aspects, at least one In is 2. In some aspects, R5 is selected from hydrogen, phenyl, and phenyloxy.

In some aspects, the polyamine compound is selected from Ra Ra Ra and a salt thereof.

In some aspects, the polyamine nd is selected from: "'2N[(H20)2J'”\|]2l’|2C CHleH(CH2)3]2NH2 Ph "'2N[(H20)3|‘”\|]2|‘|2C CH2)3]2NH2 2C)3HN]2H20 CHleH(CH2)3]2NH2 (CH2)3]2NH2 and a salt thereof.

In some aspects, the polyamine compound is selected from: H2N[("'2C)3HleHzC CHleH(CH2)3]2NH2 I'I2N[(H20)3HN]2H2C CH2[NH(CH2)3]2NH2 H2N[(H2C)3HN]2H2C CH2)3]2NH2 CH2[NH(CH2)3]2NH2 and a salt thereof.

[0148] In some aspects, the polyamine compound is selected from Ra Ra Ra Ra ,TkA‘t H’K9 ‘TJW s/KAB ArkA4 Ra R31\iL1J\ 'A7 Raj]\A3’ 1 '11] A]5A: A3 A6! L1 AG’ Raj]\A:’JA\ L1 A6a Ra Ra R8J\A3J\LJ\A6TJ§A4 pf/gfiRa8and a salt thereof; wherein each AIl member is an independently selected CR5.

In some s, the polyamine compound is selected from Ra Ra Ra RakA:/JA:L1J\ fl1/J\A45'A\\A89 J11 /J\A4 &A8 “1‘7 ’J\ 1 ' A7 A6 Ra A3 L1 A6 and a salt thereof; , , wherein each AI1 member is an ndently selected CR5.

In some aspects, the polyamine compound is selected from Ra Ra Ra Ra Ra : Ra Ra : Ra L1 R3 L1 R5 L1 Ra L1 R5 URS URa Ra R5 and a salt thereof.

In some aspects, Ra is an independently selected group of Formula VII: V11.

In some aspects, R5 is hydrogen. In some aspects, L1 is selected from a bond and O.

In some aspects, Rm is —CH2-. In some aspects, m is 1. In some aspects, each m is 1. In some aspects, at least one m is 1. In some aspects, each m is 2. In some aspects, at least one mis2.

[0153] In some aspects, Ra is —CH2[NH(CH2)n]pNH2; each n is an integer independently ed from 3 to 12; and each p is an integer independently ed from 1 to 3.

In some aspects, the polyamine nd is selected from H2N[(H20)3HNl2HzC CHleH(CH2)3]2NH2 H2N[(H20)3HN]2H2C CHleH(CH2)3]2NH2 H2N[(H2C)3HN]2H2C CHleH(CH2)312NH2 CH2[NH(CH2)3]2NH2 H2N[(H2C)3HN]2H2C CH2[NH(CH2))3]2NH2 CH2[N H((CH2)3]2NH2 "'2N[("'2C)3HleHzC OO CHleH(CH2)3]2NH2 CHleH(CH2)3]2NH2 OiPr H2N[(H20)3HN]2H2C O CHleH(CH2)3]2NH2 )3HN]2H2C CH2[NH(CH2)3]2NH2, and a salt f.

In some aspects, R5 is hydroxyl, alkoxy, lkoxy, heterocycyloxy, y, arylalkyloxy, heteroaryloxy, or heteroarylalkyloxy. In some aspects, R5 is hydroxyl, alkoxy, cycloalkoxy, heterocycyloxy, arylalkyloxy, heteroaryloxy, or heteroarylalkyloxy. In some aspects, R5 is hydroxyl, alkoxy, cycloalkoxy, or arylalkyloxy. In some aspects, R5 is hydroxyl, alkoxy, or cycloalkoxy. Preferably, R5 is alkoxy.

In some aspects, Ra is -CH2[NH(CH2)n]pNHR4; each n is an integer independently ed from 3 to 12; and each p is an integer independently selected from 1 to 3.

Preferably, R4 is alkyl, cycloalkyl, or arylalkyl; more preferably, R4 is alkyl.

In some aspects, Ra is -CH2[NH(CH2)n]pNHR4; each n is an integer independently selected from 3 to 12; and each p is an integer independently ed from 1 to 3.

Preferably, n is 3. More preferably, R4 is not hydrogen.

[0158] In some aspects, the polyarnine compound is: or a salt thereof. Preferably, R4 is isobutyl.

In certain aspects, the invention provides a polyamine composition for treatment of biofilms, the composition comprising, consisting essentially of, or consisting of a polyamine compound of formula or a salt f; wherein: each Ra is an independently ed group of Formula I: A1, A2, A3, A4, A5, A6, A7, A8, and A9 are each an A11 member independently selected from N, CR3, and CR5; wherein at least one A11 member and at most five AIl members are independently selected CRa; each R121 and R1b is a member ndently selected from hydrogen, fluoro, alkyl, and fluoroalkyl; each Rza, R21), RR, and R2d is a member independently selected from hydrogen, alkyl, alkyl, aryl, and arylalkyl; each Rm is a member independently ed from —CR2aR2b- and -C(R2a)(R2b)-L2- C(R2°)(R2“)-; each m is an integer independently selected from 1 to 20; each L1 is a member independently selected from a bond and -O-; each L2 is a member independently selected from a bond, -O-, and -NR4-; each R3 is a member independently selected from -Z1-R4, -Zl-Y1-R4, and -Z1-Y1-Y2- each R4 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, and arylalkyl; or, alternatively, for a -N(R4)2 group, one of the two R4 in the group is a member ed from -(CO)OR6a-, -(CO)N(R6a)(R6b), and -C(NR63)N(R6b)(R6C); each R5 is a member independently ed from hydrogen, alkyl, hydroxyl, alkoxy, alkylamino, aryl, aryloxy, cyclyl, halo, fluoroalkyl, fluoroalkyloxy, heteroaryl, arylalkyl, arylalkyloxy, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; each Y1 and Y2 is an independently selected group of Formula IA: 1 0 IA each Z1 and 22 is a member independently selected from NR4 and 0; each R6a, Réb, and R60 is a member independently selected from hydrogen and alkyl; wherein if R4 is —C(O)OR6a, R6a is alkyl; and wherein the biocidal polyamine compound comprises at least two primary or secondary amino groups.

In some aspects, the composition ses a polyamine nd of formula 1%,“ 5’A§A89 €12 J\ q I A\A3/ 2A7 L1 A6 or a salt thereof.

In some aspects, each A1, A2, A3, A4, A5, A6, A7, A8, and A9 are each an A11 member independently selected from N, CRa, and CR5 ; wherein at least one AI1 member and at most five AI1 members are independently selected CRa.

In some aspects, each R1&1 and R1b is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl.

In some aspects, each Rza, RZb, R20, and R2d is a member independently selected from hydrogen, alkyl, lkyl, aryl, and arylalkyl.

[0164] In some aspects, each Rm is a member independently selected from —CR2aR2b- and _C(R2a)(R2b)_L2_C(R20)(R2d)_ .

In some aspects, each R6a, R61”, and R60 is a member independently selected from hydrogen and alkyl; wherein if R4 is R6a, R621 is alkyl.

In some aspects, each L1 is a member independently selected from a bond and -O-; and each L2 is a member independently selected from a bond, -O-, and -NR4-.

[0167] In some aspects, each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, mino, aryl, aryloxy, heterocyclyl, halo, fluoroalkyl, fluoroalkyloxy, heteroaryl, arylalkyl, arylalkyloxy, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl.

In some aspects, each R6a, R61”, and R60 is a member ndently selected from hydrogen and alkyl; wherein if R4 is —C(O)OR6a, R6a is alkyl.

[0169] In some aspects, the invention provides a polyamine compound selected from: (a) a compound of Formula IC: Rq R3 Rm R1 ( N\ RQNWLT‘KNMLNH /—NH HN q H I n H R1 R” N/\l/CDRHR)p R1 (b) a compound of Formula ID: Rm’“ R1 R3\ Ea NR3R§N~(27LQN'IR4 4 1 hair“: I R R R R \/j— W) P /\’/K R1 RP R5 (R6)Z or a salt thereof; 1 5 wherein: each R1, Rm, Rn, RP, and Rq is independently a substituent independently selected from hydrogen, alkyl, l, alkynyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl; each R2 is independently a tuent independently selected from hydrogen, alkyl, alkoxy, alkylamino, alkenyl, alkynyl, aryl, aryloxy, arylamino, halo, haloalkyl, heteroaryl, aryloxy, heteroarylamino, kyl, alkylaryloxy, rylamino, heteroarylalkyl, heteroalkylaryloxy, heteroalkylarylamino, Rm Rm R1 R1 R: N?“ R: WT» N N n if R1 R1 and R1 R1 Rn each 2 is is independently an integer independently selected from 1 to each m, n, p, and q is independently an integer independently selected from 1 to 20; each R3 is independently a substituent independently selected from hydrogen, each R4 is independently a substituent independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, R1 R1 and R1 R1 R" , ; and each R5 is independently a substituent independently selected from alkyl, , alkylamino, alkenyl, alkynyl, aryl, aryloxy, arylamino, halo, haloalkyl, heteroaryl, heteroaryloxy, heteroarylamino, arylalkyl, alkylaryloxy, alkylarylamino, heteroarylalkyl, heteroalkylaryloxy, hetReroalkylarylamino, Rm Rm R1 R1 3 (R6)Z R1 m a R1 09% RN N N n f R4 R1 Rnn R3NY R1 R1 R1 R1 R” I/J and , , Rq R3 RmmFlz3 R1 R\WWW“!3 NR3R§N4QtNR4’ 7:]— Hop R1 R4 R1 R” R1 RD 4 R6)z ;and each R6 is independently a substituent independently selected from hydrogen, alkyl, alkoxy, alkylamino, alkenyl, alkynyl, aryl, y, arylamino, halo, kyl, heteroaryl, heteroaryloxy, heteroarylamino, arylalkyl, ryloxy, rylamino, arylalkyl, heteroalkylaryloxy, and heteroalkylarylamino.

In some aspects, the polyamine compound is selected from: (a) a compound of Formula IE: (b) a compound of Formula IF: wherein: each R1, Rm, R“, R”, and Rq is independently a substituent ndently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, aryl, arylalkyl, and heteroarylalkyl; each R2 is independently a substituent independently ed from hydrogen, alkyl, alkoxy, alkylamino, alkenyl, alkynyl, aryl, aryloxy, arylarnino, heteroaryl, heteroaryloxy, arylamino, arylalkyl, alkylaryloxy, alkylarylarnino, heteroarylalkyl, heteroalkylaryloxy, heteroalkylarylamino, R1 M22, R1 09:» N N n ,6 R1 R1 and R1 R1 Rn each m, n, p, and q is independently an integer independently selected from 1 to 20; each R3 is independently a substituent independently selected from en, Rm Rm R1 R1 R1 my R1 991% N N “a“ R1 R1 ,and R4 R1 Rn each R4 is independently a substituent independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, Rrn Rm R1 R1 Riwa R1 mi: N N “z“ R1 R1 and R1 R1 R” , ; and each R5 is independently a substituent independently selected from alkyl, alkoxy, alkylamino, alkenyl, alkynyl, aryl, aryloxy, arylamino, heteroaryl, heteroaryloxy, heteroarylamino, kyl, alkylaryloxy, alkylarylarnino, heteroarylalkyl, alkylaryloxy, heteroalkylarylamino, In some aspects, the ine compound is CH2[NH(CH2)3]2NH2 In still some aspects, R2 is hydrogen.

In yet some aspects, R5 is aryl or aryloxy.

In still yet some aspects, R5 is In some aspects, the ine compound is 2NA<\/\NH HN/\/)~NH2 2 2 or a salt thereof.

In some aspects, the polyamine compound is umwm CH2[NH(CH2)3]2NH2 or a salt thereof.

In some aspects, the polyamine compound is H NNN NwlNH or a salt thereof.

In some aspects, the polyamine compound is HNNN N/\/>NH of) or a salt thereof.

In some aspects, the invention provides a polyamine compound of Formula LP or a salt thereof: E Elwfw“Nww-MMWNRW) F «J H 2', wherein: M is selected from: (a) a C1-C13 alkyl, alkenyl or alkynyl group, and (b) —(L—NH)Z—L—, where L is a C3-C13 alkyl, alkenyl or alkynyl group, and where z is an integer from zero to 6; x is an integer from 1 to 6; and R1 and R2 each is independently selected from hydrogen and C1—C3 alkyl group.

[0180] In some aspects, the polyamine compound of a I-P may se a hydrophobic phenyl group and up to six hydrophilic polyamine chains. Each of the hydrophilic polyamine chains may be the same, or some of the hydrophilic polyamine chains may be same, or none of the hydrophilic polyamine may be the same. Additionally, each hilic ine chain may comprise a neutral amine, a cationic ammonium salt, or both.

In some aspects, the ine compound may comprise a compound of Formula II-P or a salt f, wherein n is an integer from 1 to 13, x is an integer from 1 to 6, and R1 and R2 each may independently be hydrogen or a C1-C3 alkyl group. .» \ ff {”9 ”'22" "'" H {V ‘X \W?’ II-P In some aspects, the polyamine compound may comprise a compound of Formula III-P or a salt thereof, wherein n is an integer from 1 to 13, and the hobic phenyl group is substituted with two hydrophilic polyamine chains at the ortho-, meta-, or para-position.

,A r’ I; \W‘Nwwrcmnmwzifi Kg; H I III-P In yet some aspects, the polyamine compound may comprise a compound of Formula IV-P or a salt thereof, n each m and n is an independently ed integer from 1 to 13.

H N “Mi, CH“RpmN”W?.»»~”’"\‘:§ “N } Nmiitfifiér _______NH H a H l 5 IV-P In some aspects, m is 12 (i.e., compound IV-l or a salt thereof). In some alternative aspects, m is 6 (i.e., compound IV-2 or a salt thereof).

In some aspects, the polyamine compound may comprise a compound of Formula V-P or a salt thereof, wherein n is an integer from 1 to 13, and the three hydrophilic polyamine chains may be at any position on the hydrophobic phenyl group. ,- f l FT’WWEWHJB In some aspects, the polyamine compound may comprise a compound of Formula VI-P or a salt thereof, n each m, n and p is an independently selected integer from 1 to l3: and each R1 and R2 may independently be hydrogen or a C1—C13 alkyl group.

*RERN«-w<::u2}m--wn‘ #fN~ VI-P In some aspects, m, p, and n are 3 (e.g., compound VI-l). In some aspects, m, p, and n are 6 (e. g., compound VI-2).

[0188] In some aspects, the polyamine compound may comprise a compound of Formula VII-P or a salt f: foltx 'L x «\ A. §/‘\ xx {ii saw [,, ‘ NH’ “v“ “m ‘v” “ New?) ”If H 3" xxifigx VII-P wherein: x is an integer from 1 to 6, y is an integer from zero to 6, each R1 and R2 is independently ed from hydrogen and C1-C13 alkyl; and wherein the substituted hydrophilic polyamine chains may be at any on of the hydrophobic phenyl group.

[0189] In some aspects, the polyamine compound may comprise a compound of Formula VII-l or a salt thereof, wherein y is an integer from zero to 6, and the two hydrophilic polyamine chains may be at the ortho-, meta-, or para- position of the hydrophobic phenyl group .

VII-1

[0190] In some aspects, the polyamine compound may comprise a compound of Formula VII-2 or a salt thereof. x \f‘Ax (“xN” fix. {,1«\‘c.er’ié.‘\\\II» \ .-”(\\,,xx \ /J\ “/0.

”EN N N NH f N \x XNHw H H kg? H V: VII-2 In some s, the polyamine nd may comprise a compound of Formula VII-3 or a salt thereof, wherein y is an integer from zero to 6, and the three hilic polyamine chains may be at any position of the hydrophobic phenyl group. [if \f‘9"“lew”f‘v‘mwi:\’”’A\NRIR§)3 VII-3 In yet some aspects, the polyamine compound is a compound of Formula VII—4 or a salt thereof, wherein f, g and h are each an independently selected integer from zero to 6. -3mmL a» A w“ J“ ”V x 9 «~ “ii \V 14mm H j \NH”T\\ \‘NHE»!\NR‘RZ» \V’fi’ LNH "{\V’”fl\ f]05V”NR1 R2 VII-4 In some aspects, the polyamine compound is a compound of Formula VII-5 or a salt thereof: "a“ \\Niki} WM» “No, I -' ' r ”(6135533. f .~ 3x{SHEER K ”MS“ N" 2 “NR RNT H1 ‘“‘§. 3‘ i w E i ‘ E #1:: f ‘X VII-5 wherein: a is an ndently selected integer from 2 to 13, x is an independently selected integer from 1 to 6, y is an independently selected integer from zero to 6, each R1 and R2 is independently selected from en and C1-C13 alkyl, and the substituted hydrophilic polyamine chains may be at any position of the hydrophobic phenyl group.

In some aspects, the polyamine nd is a compound of Formula VIII—P or a salt thereof: ’w‘lj"\\\ i“, ii 1 gill ”’i’fli‘w‘x '9‘ ‘ '“ k \ Nm-=-M~WWNR R‘ § § H 3 xxgfififirw VIII-P wherein: M may be selected from: (a) C1-C3 alkyl, alkenyl or alkynyl group, (b) —(L—NH)Z—L—group, wherein L is C3-C3 alkyl, alkenyl or alkynyl group, and z is an integer from zero to 6; x is an r from 1 to 6; and R1 and R2 each may independently be hydrogen or a C1-C1; alkyl group.

The polyamine compound of general Formula VIII-P may comprise a hydrophobic phenyl group and up to six hilic polyamine chains, each of the hydrophilic polyamine chains connecting to the hydrophobic phenyl group Via an amide functional group. Each of the hydrophilic polyamine chains may be the same, or some of the hilic polyamine chains may be same, or none of the hilic polyamine chain may be the same.

Additionally, each hydrophilic polyamine chain may comprise a neutral amine, a cationic ammonium salt, or both.

In some aspects, the polyamine compound is a compound of Formula VIII-l or a salt thereof, wherein n is an r from 3 to 13, and the two hydrophilic polyamine chains may be at the ortho-, meta— or para- position of the hydrophobic phenyl group. ti. {3 "ME N " ”((3425$le is3) Ki'x j H i E: 9’13: \z’fif VIII-1 In some aspects, the polyamine compound is a compound of formula VIII-2 or a salt thereof, wherein y is an r from zero to 6, and the two hydrophilic polyamine chains may be at the 0rth0-, meta—, or para- position of the hydrophobic phenyl group. 73,,» N5; ’ i Emmi?x f» "N N” ,N “f \N3‘ \NH.::) N2 VIII-2 In some aspects, the polyamine nd is a compound of Formula VIII-3 or a salt thereof. {j} HN"»\W’” ” \[lfklfkg N:N ' {\N‘} x\\¢§§ {"l N VIII-3 In some aspects, the polyamine compound is a compound of Formula VIII—4 or a salt thereof: VIII-4 a is an integer from 2 to 13, x is an r from 1 to 6, y is an integer from zero to 6, each R1 and R2 is independently selected from the group hydrogen and a C1-C13 alkyl group, and the substituted hydrophilic polyamine chains may be at any position of the hydrophobic phenyl group.

In some apects, the ine compound is a compound of Formula I-P, Formula VIII-P, or a salt thereof, wherein M, R1 and R2 are as described herein, and x is an integer from 1 to 6. The polyamine compound comprises a hydrophobic phenyl group and at least one hydrophilic polyamine chain.

[0201] In certain aspects, the compounds of the t invention are antimicrobial and provide triple action against bacteria and biofilms. ageously, the antimicrobial compounds of the t invention have specific activity against biofilms.

Organisms residing in biofilms present a complex extracellular matrix of polysaccharides (exopolysaccharides) and proteins. As a result of this complex matrix, nutrient-limiting conditions exist that alter the normal or planktonic metabolic state. These conditions reduce the efficacy of ional antibiotic agents, rendering them up to l,000x less active against biofilms. A hallmark of these exopolysaccharides is the presentation of acidic es from repeated glucoronic acid motifs and pyruvate derived acetals. A recent study by Losick and kers demonstrated that the simple polyamines spermine and norspermidine (A) were naturally occurring inhibitors of biofilm formation, endogenously produced at high concentrations (5 0-80 uM) in response to nutrient limiting conditions and waste accumulation in mature pellicles (Kolodkin-Gal, I. et al., A self- produced trigger for biofilm disassemby that targets exopolysaccharide. Cell 149 (2012)). In this study, they were able to demonstrate that norspermidine could t biofilm formation at 25 uM and showed that, at similar concentrations, it could disperse the exopolysaccharide component of the matrix but not the protein ent. Interestingly, dine was only active at much higher concentrations (~l mM) leading them to propose a ale for this activity in the ability of the polyamines to engage the acidic es in the matrix at regular intervals (B).

In certain aspects, compounds of the present invention having increased numbers of chains, produce a more effective compound against A. baumanniz’. For example, compounds with four polyamine chains can be generated with Pd(II) mediated dimerization of 5- bromoisopthalaldehyde followed by reductive amination ().

[0204] In certain s, the compounds of present invention combine a hobic backbone with a cationic tail that have the functionality to inhibit biofilm formation, disrupt established biofilms, and kill the emerging planktonic bacteria. Certain of the compounds of the ion are set forth in FIGS. 27 and 28.

In a particular embodiment, the polyamine compound may comprise a hydrophobic I5 moiety head and at least one hydrophilic moiety tail comprising a polyamine group. When the polyamine compound comprises more than one hydrophilic moiety tails, the hydrophilic moiety tails may be the same, or alternatively, the hydrophilic moiety tails may be different.

As discussed above, the exemplary polyamine nds shown herein are not ed to be ng.

SYNTHESIS The synthesis of diaminopropane substituted backbones is straightforward and results in CZ-4, 12, 32 () from the known mono-Boc protected diaminopropane and commercially available aldehydes (benzaldehyde, isopthalaldehyde or 1,3,5— benzenetricarboxaldehyde). This three-step synthetic procedure ds Via ive amination (Baxter, E. W. & Reitz, A. B. Reductive Aminations of Carbonyl nds with Borohydride and Borane Reducing Agents. Org Reac l, 59 (2004)) and acidic removal of the Boc group. The norspermidine series (e.g., CZ-7, CZ-25, and CZ-52) can be prepared in a similar manner from the mono-Boc protected norspermidine (). No purification is required until a final recrystallization of the HCl salt, which has allowed easy preparation of these compounds on larger scale.

In some embodiments, the polyamine compound may be produced by the reductive amination method of General tic Scheme 1. l Synthetic l + H N—(CR2n yH—(CRz)—nr{J—R1 —> Step1 y= 0-2 n=2-20 R1: 800, alkyl acy/ su/fony/ \ 1L Y = H, formyl R2__ H, Me, [> I,~N\ NH X = H, halo, formyl, aryl, R1’ R3 _ "" R3 = H formylphenyl, heteroaryl, 0R4 R4 = H, alkyl, benzyl, alkylaminoalkyl R1—E:(CR2)-n—N CR2)-N Hn—(CR2 yH-(CR2)—nN-—R1 Step 2 NIXCNWNN[QWNWig:1N1;.N; 0(NVNN —total # of basic amines Reagents: (a) MeOH, 3 A molecular sieves, rt, 1-28 h; NaBH4; (b) HCl, MeOH, rt 1-4 h.

The General Synthetic Schemes included herein set forth methods of preparing some compounds within the aspects and embodiments sed herein. In some alternative aspects, these methods can be used to produce a compound of the one of the s and embodiments disclosed herein. In some alternative aspects, these methods can be used to produce a precursor or starting material for producing such a compound.

In General Synthetic Schemes l, 2, 3, and 5, “CR2” indicates a methylene with two substituents, which may be hydrogen, methyl (or, alternatively, lower alkyl), or spirocyclopropane (z'.e., the two tuents are the spirocyclic ring).

In some embodiments, the polyamine compound may be produced by the acylation method of l Synthetic Scheme 2.

General Synthetic Scheme 2 O 0 CI CI H H H a + H N CR2 N CR2 N—R1 —> n " Step1 x y = 0-2 n = 2-20 R1 = Boc, alkyl, acyl, sulfonyl X = H, haloaryl, R2 = H’ Me, [> ar R3 = H yI, OR4 R4 = H, alkyl, benzyl, alkylaminoalkyl O O H H H H b n H H n n Step2 O O H H H H R1—N CR2 N CR N N CR2 N CR2 N-R1 '(HCl)z n H H n n z = O — W W = total # of basic amines Reagents: (a) TEA, CHzClz, rt, 1-28 h; (b) HCl, MeOH, rt 1-4 h.

In some embodiments, the polyamine compound may be produced by the reductive amination method of General Synthetic Scheme 3.

General Synthetic Scheme 3 H H H H a H2N CR2 N CR2 N N CR2 N CR2 NH2 _> n n n n Step 3 y y H H H H H H b N CR2 N CR2 N N CR2 N CR2 N—CH2R1 —> n n n n Step4 y y H H H H H H R1HZC-N CR2 N CR2 N N CR2 N CR2 N-CH2R1-(HCI)Z n n n n y y z = O - W X W = total # of basic amines Reagents: (a) R1CHO, MeOH, 3 A molecular sieves, rt, 1-28 h; NaBH4; (b) HCl, MeOH, rt 1- 4 h.

In some ments, the polyamine compound may be produced by the reductive amination method of General Synthetic Scheme 4.

Ge_v—neralS nthetic Scheme 4 l\a/R4 OH Step 1 Step 2 O‘R4 H H —> _> + H Step 3 N‘(CR2H’:H4(CR2>—nN——R1 Step 4 0eR4 y= 0-2 n= 2-20 R1 = Boc, alkyl, acyI, yl e, [> R3=H R4 = H, alkyl, benzyl, alkylaminoalkyl H H H H H H e R1—N CR2 N CR N N CR2 N CR2 N—R1 —> n n n Step5 y y R1_N CR2 nN CR2 CR2” CR2 nN—R1 o (HC|)Z Z=0-W W= total # of basic amines Reagents: (a) CH3CN, Cs2C03, rt, 16 h; (b) THF, LiAlH4, rt, 8 h; (c) PCC, CH2C12, rt 1-4 h; (d) MeOH, 3 A lar sieves, rt, 1-28 h; NaBH4; (e) HCl, MeOH, rt 1—4 h.

[0214] In some aspects, the R4CH21 can include a different leaving group for the nucleophilic substitution reaction of (a). Other leaving groups useful for phenol alkylation include Br, Cl, tosylate, mesylate, triflate, and the like.

The precursor compounds shown below were made by the method of Steps 1 to 3 in the General Synthetic Scheme 4 above. 5-(Cyclohexylmethoxy)isophthalaldehyde: 1H NMR (300 MHz, CDC13) 8 ppm .02 (s, 2H), 7.91 (s, 1H), 7.61 (s, 2H), 3.84 (d, J: 6.0 Hz, 2H), 1.87-1.67 (m, 6H), 1.31- 1.03 (m, 5H). 13C NMR (75 MHz, CDC13)8 ppm 191.4, 160.9, 138.6, 124.3, 120.2, 74.6, 37.9, 30.1, 26.7, 26.0. 0 0 \OJ\/ 5-(2-Ethy1butoxy)1sophtha1aldehyde: 1H NMR (500 MHz, CDC13) 8 ppm 10.01 (s, 2H), 7.90 (s, 1H), 7.62 (s, 2H), 3.94 (d, J= 5.5 Hz, 2H), 1.68 (hept, J= 6.5 Hz, 1H), 1.46 (dec, .1: 6.5 Hz, 4H), 0.91 (t, J = 7.5 Hz, 6H). 13(2 NMR (125 MHz, CDC13) 8 ppm 191.1, 160.7, 138.5, 124.1, 120.0, 71.1, 40.9, 23.5, 11.3. o’ \0 5-(Hydroxymethy1)1sophthalaldehyde: 1H NMR (500 MHZ, CDC13) 8 ppm 10.09 (s, 2H), 8.27 (s, 1H), 8.15 (s, 2H), 4.88 (s, 2H), 2.54 (br s, 1H). 13c NMR (125 MHz, 8 ppm 191.4, 143.7, 137.4, 132.8, 130.3, 63.9.

[0219] In some embodiments, the polyamine compound may be produced by the method of Scheme 5, wherein R1 may be hydrogen or a C1-C13 alkyl group.

Scheme 5 HzN“"'WMW‘-NHR" {‘1} C} l(:BQC]20 ’x’N\ f H ‘ ‘ k“ ”ii-“ME: CH) + HRNMN]2...... {x} R 3806 ,i ' 3‘ {2] ~55” “i MeOH. moiacuiar sieve 24 News“ fe/‘ctw‘ 1” psi wgm]” ’f%\ w [ Acid, MeoH waif ”‘x A - .............................

K J J, H x ] NMM»-NHR j- (g; (A H k \frr \Wgw {4] LA The method of Scheme 6 may include reacting a polyamine of formula [1] with di-t- butyldicarbonate nd [(Boc)20] to protect at least one terminal amine group of the polyamine [1], while leaving at least one terminal amine group of the polyamine [1] unprotected. The resulting polyamine of formula [2] having at least one Boc-protected terminal amine is reacted With a substituted benzadehyde of formula [3]. Then, the resulting product is reduced, such as by a hydride reducing agent (e. g., NaBH4 or LiAlH4) to provide a corresponding polyamine ate of formula [4] having the terminal amine group on at least one hydrophilic polyamine chain Boc-protected. The Boc-protected terminal amine group is then deprotected, such as by acid hydrolysis, to provide the polyamine compound of Formula I-A comprising a hydrophobic phenyl group and at least one hilic ine chain.

In some embodiments, the ine compound of formula VIII-P may be produced by the method of Scheme 6, wherein R1 may independently be hydrogen or C1-C13 alkyl group.

Scheme 6 HgflmeNHz gqancho (I: i v” $yfi(,g~—cg) l HENMMWNRT‘BDC fl " X {2} \5?’ E5} NEt3, salami E «j—’i’c\gwwgum-mm“Bo;x ‘v [6] Acid, MeOH fffkkt§w :"{3‘\\.

Ll xl ““NwM-~«~«N2—§R*)H ‘ijgfif VIII-P The method of Scheme 7 may include reacting the tuted benzoyl chloride of formula [5] with a polyamine of formula [2] having at least one terminal amine group protected with a Boc-protecting group, in a presence of base such as triethylamine (NEt3) to provide a corresponding substituted benzyl amide of formula [6]. The compound of formula comprises a hydrophobic phenyl group and at least one hydrophilic polyamine chain having a Boc-protected terminal amine group. Then, the Boc-protected terminal amine group ofthe hilic polyamine chain is deprotected, such as by acid hydrolysis, to provide the polyamine nd of Formula VIII-P.

The polyamine compounds exhibit unexpectedly superior ability in treating biofilms, compared to most known antimicrobial compounds. The microorganisms in the biofilm community may be eradicated ively and rapidly such that they have minimum, if any, opportunity to upregulate their defense ism and develop resistance against the polyamine compound. Without being bound by any theory, it is believed that the unexpectedly superior biofilms treatment is due to the beneficial synergistic of hobicity and hydrophilicity of the polyamine compound. It is believed that the hobic moiety of the ine compound facilitates the dispersion of the microorganisms in biofilms, while the hydrophilic polyamine moiety provides an antimicrobial effect on the dispersed microorganisms.

The polyamine compounds may eradicate the biofilms, reduce the formation of biofilms, or inhibit the formation of ms. The hydrophobicity and hydrophilicity of the polyamine compounds may be designed by tailoring the hydrophobic moieties and hydrophilic es of the polyamine compounds such that the desired level of antimicrobial effect on the biofilms may be achieved. One d in the arts recognizes the ters controlling the hydrophobicity/hydrophilicity effect, and, therefore, may readily modify the teachings of t disclosure to produce various polyamine compounds without departing from the scope of present sure. As non-limiting examples, the hydrophilicity effect of the polyamine compound may be modified by varying numbers of the hydrophilic ine chains in the polyamine compound, numbers of amine groups in the hilic polyamine chains, numbers of s between amine groups in the hydrophilic polyamine chains, etc.

As non-limiting examples, the hydrophobicity effect of the polyamine compound may be modified by g positions of the hydrophilic polyamine chains on the hydrophobic groups, altering chemical structures of the hydrophobic groups to other known non-polar functional group, etc.

The polyamine compounds may exhibit enhanced antimicrobial effect on ms comprised of Gram-negative or Gram-positive bacteria. The polyamine compounds may exhibit enhanced antimicrobial effect on biofilms consisting of mycobacteria.

In one embodiment, the antimicrobial composition may comprise a polyamine compound and at least one additive. Various additives may be used for the antimicrobial composition. By way of non-limiting examples, the additives may further enhance the dispersion of microorganisms in biofilms, impart the antimicrobial effect against the dispersed microorganisms, facilitate the application/administration of the antimicrobial composition to the biofilms, improve the stability of the antimicrobial composition, control the release/application rate of the antimicrobial composition to the biofilms, etc. Non- ng examples of additives for further enhancing the antimicrobial effect may be e and other bactericide. By way of non-limiting examples, the additives for facilitating the administration of the antimicrobial composition may e a pharmaceutically acceptable carrier typically used for medical or pharmaceutical ations, an emulsifier or dispersant typically used for industrial applications.

The antimicrobial composition may be formulated to provide the desired level of crobial effect on the biofilms by selecting a polyamine compound and other ves as well as by adjusting the amount of each component in the antimicrobial composition. In some ments, the antimicrobial composition may be formulated to inhibit the formation of biofilms. In some embodiment, the antimicrobial composition may be formulated to disrupt the biofilms. In still other embodiments, the antimicrobial composition may be formulated to ate substantially all microorganisms in the biofilms.

Any suitable amount amine can be used in the compositions and methods of the invention. In general, the polyamines are used in concentrations ranging from about 1 ppm to about 100,000 ppm, or higher. The concentration of a polyamine used in a composition or method of the invention can be, for example, from about 1 to about 100,000 ppm, or from about 10 to about 10,000 ppm, or from about 100 to about 1,000 ppm, or from about 1 to about 100 ppm, or from about 1,000 to about 10,000 ppm, or from about 10,000 to about 100,000 ppm. The concentration of a polyamine can be about 1; 2; 3; 4; 5; 6; 7; 8; 9; ; 15; 20; 25; 30; 35; 40; 45; 50; 55; 60; 65; 70; 75; 80; 85; 90; 95; 100; 125; 150; 175; 200; 225; 250; 275; 300; 325; 350; 375; 400; 425; 450; 475; 500; 525; 550; 575; 600; 625; 650; 675; 700; 725; 750; 775; 800; 825; 850; 875; 900; 925; 950; 975; 1000; 1500; 2000; 2500; 3000; 3500; 4000; 4500; 5000; 5500; 6000; 6500; 7000; 7500; 8000; 8500; 9000; 9500; ,000; 12,500; 15,000; 17,500; 20,000; ; 25,000; 27,500; 30,000; 32,500; 35,000; 37,500; ; 42,500; ; 47,500; 50,000; 52,500; 55,000; 57,500; 60,000; 62,500; 65,000; 67,500; 70,000; 72,500; 75,000; 77,500; 80,000; 82,500; 85,000; 87,500; 90,000; 92,500; 95,000; 97,500; or about 100,000 ppm. Other concentrations ofpolyamines can be usefial in the compositions and methods of the ion, depending in part on factors including the specific polyamine used, the presence of potentiating agents if any, or the species ofmicroorganisms that are targeted.

In certain aspects, increasing number of polyamine chains on the hydrophobic backbone systematically increase the antimicrobial activity. For example, it is demonstrated herein that with a single chain of diaminopropane added to a benzyl backbone (CZ-4, ), the MIC (as defined by the Clinical and Laboratory Standards Institute (CLSI)) against MRSA is greater than 1,200 ug/mL, whereas two chains decrease the MIC to 300 ug/mL ) and three chains reduce it r to 45 ug/mL (CZ-32). Similarly, with a single chain ofnorspermidine attached to a benzyl backbone (CZ-7, ), the MIC against MRSA is greater than 1,200 ug/mL. By adding a second chain of norspermidine (CZ-25), the MIC is 45 ug/mL. With a third chain of norspermidine attached (CZ-52), the MIC becomes 3 ug/mL ().

This trend of ing antimicrobial activity by sing the number of polyamine chains attached to a backbone is effective for treating MRSA, but modulating this trend with increased hydrophobicity enhances the activity of CZ compounds against A. baumanm‘i. Interestingly, CZ-52, with three norspermidine chains, has greater activity against MRSA s than against A. nii, whereas CZ-58 () has greater activity (10-fold increase) against A. baumanm’z’ biofilms than MRSA biofilms (see Table 4 below).

APPLICATIONS As described herein, biofilms can also affect a wide y of biological, medical, and processing operations. Methods and treatments using a polyamine nd, or a combination of a polyamine compound with another compound, may include killing, dispersing, treating, reducing biofilms or preventing or inhibiting biofilm formation.

In one embodiment, the ion es a method for dispersing or killing a biofilm, the method comprising a step of treating the biofilm with an anti-biofllm ition, thereby effectively dispersing or killing the biofilm; n the method comprises, consists essentially of, or consists of using a polyamine compound or composition as set forth in any of the embodiments or aspects described herein.

In some aspects, the step of treating the biofilm with an anti-biofilm composition effectively disperses the biofilm.

[0234] In another embodiment, the invention provides a method for inhibiting formation of a biofilm, the method comprising a step of ng planktonic bacteria with a polyamine composition as set forth in any of the ments or s herein, thereby inhibiting incorporation of the planktonic bacteria into the biofilm.

In certain s, the method of killing, dispersing, dislodging, ng, or reducing biofilms, or preventing or inhibiting biofilm formation, includes contacting the biofilm with an effective amount of a composition of the present ion.

In some aspects, the formation of a biofilm is inhibited. In other aspects, a previously formed biofilm is dispersed. In still other aspects, substantially all of the cells comprising a biofilm are killed.

In some embodiments, the invention provides a method of killing, dispersing, treating, or reducing biofilms, or preventing or inhibiting biofilm formation, the method comprising contacting a biofilm or a surface having a biofilm disposed thereon with an effective amount of a polyamine compound.

In some aspects, a surface comprises a medical device, a wound dressing, a contact lens, or an oral device. In some aspects, the medical device is selected from a clamp, forceps, scissors, skin hook, , needle, tor, , drill, chisel, rasp, saw, er, orthopedic device, artificial heart valve, prosthetic joint, voice prosthetic, stent, shunt, pacemaker, surgical pin, respirator, ventilator, and an endoscope and combinations thereof.

In some s, the method described herein comprises, consists essentially of, or consists of using the polyamine compound or composition described in any of the embodiments or aspects herein.

In some aspects, the invention provides a method that ses, ts essentially of, or consists of using a polyamine compound or composition from any of the embodiments or aspects described herein.

In some embodiments, the ion provides a method for dispersing or killing a biofilm, the method comprising a step of treating the biofilm with an anti-biofilm composition, thereby effectively sing or killing the biofilm; wherein the anti—biofilm composition comprises, consists essentially of, or consists of a polyamine compound selected from A2 A3 Ra Ra Ra Y 1&A5 1 A§89 L1 A9 1\ 4 5A\\89 \8 I I l l‘ I A A I21&(L 4 ls i i \E '7 A 2A A 2A A / :A A ,A \A3 \A3 \A6 \A3 L1 A6 \AB and a salt thereof; , , , , wherein: each R21 is a member independently selected from m R10 R1a 1b R181 1b EA< RZa R1d R28 z1J<R2b R2 sz N_kR R20 ( Rm%R2d R1 ( Rm%R2d m R3 m and R3 ; A1, A2, A3, A4, A5 , A6, A7, A8, and A9 are each an A11 member independently selected from N, CRa, and CR5; or, alternatively, a pair of adjacent AI1 members join to form an independently selected aryl, cycloalkyl, heterocyclyl, or cycloaryl ring that is fused with an A11 ring at the pair’s AI1 ring ons; wherein at least one AI1 member and at most five AI1 members are an independently selected CR3; each R13, Rlb, R”, and R1d is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl; or, alternatively, an R1a and an R1b join to form an oxo group; each Rza, sz, R2°, de, R26, and R2f is a member independently selected from en, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl; alternatively, a pair of R2 members from the same Ra group independently selected from the group R2a and R21), R20 and RN, and R2‘3 and n to form a member independently selected from spirocycloalkyl, spiroheterocycyl, and oxo; or, atively, an R2a and an R2c from the same Ra group join to form a ring independently selected from cycloalkyl and heterocycyl; each Rm is a member independently ed from -CR2aR2b-, -CR2CR2d-, -C(R2a)=(R2b)-, -cc-, and -C(R23)(R2b)-L2-C(R2c)(R2d)-; each m is an integer independently selected from 1 to 20; each L1 and L2 is a member independently selected from a bond, —o—, -C(O)O-, -NR4-, -NR4C(O)-, and -C(O)NR4-; each R3 is a member independently selected from —Zl-R4, -Zl-Y1-R4, —zl—Y1—Y2—R4, and —zl—Y1—Y2—Y3—R4; each R4 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, kyl, cycloalkylalkyl, and heteroarylalkyl; or, alternatively, for an -N(R4)2 group, one of the two R4 in the group is a member selected from -(CO)OR63-, -(CO)N(R63)(R6b), and -C(NR63)N(R6b)(R6°); or, alternatively, for an - N(R4)2 group, the two R4 groups join to form a cyclic ring; each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, aminoalkoxy, alkylamino, alkylaminoalkoxy, alkenyl, alkynyl, aryl, aryloxy, arylamino, cycloalkyl, cycloalkoxy, cycloalkylalkoxy, cycloalkylamino, cycloalkylalkylamino, cyclyl, heterocycyloxy, heterocycylamino, halo, haloalkyl, lkyloxy, heteroaryl, heteroaryloxy, heteroarylamino, arylalkyl, arylalkyloxy, arylalkylamino, heteroarylalkyl, arylalkyloxy, heteroarylalkylamino, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; each Y1, Y2, and Y3 is an independently selected group of Formula IA: each Z1 and Z2 is a member independently selected from —N(R4)—and —O—; and each R621, R61), and R60 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, arylalkyl, heteroarylalkyl, and cycloalkylalkyl; or, alternatively, two R6n s R621 and R6b or R621 and R60 join to form a heterocycyl ring; wherein the polyamine compound comprises at least two primary or secondary amino groups.

In some aspects, each R4 is a member ndently selected from en, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl; or, atively, for a -N(R4)2 group, one of the two R4 in the group is a member selected from -(CO)OR6a-, (CO)N(R“)(R""), and —C(I~IR“)N<R6")(R6°); each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, alkylamino, alkenyl, alkynyl, aryl, aryloxy, arylamino, cycloalkyl, lkoxy, cycloalkylamino, heterocyclyl, heterocycyloxy, heterocycylamino, halo, haloalkyl, fluoroalkyloxy, heteroaryl, heteroaryloxy, heteroarylamino, arylalkyl, arylalkyloxy, arylalkylamino, heteroarylalkyl, heteroarylalkyloxy, heteroarylalkylamino, yalkyl, lkyl, and alkylaminoalkyl; and each R6a, R61), and R60 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, cycloalkyl, and heteroarylalkyl; or, atively, two R611 members R621 and R6b or R621 and R60 form a heterocycyl ring.

In some aspects, the method has the proviso that the polyamine compound is not of Formula IB: \N4 NMN/R4 KL F'a4 F'z4 J) R4 R4 wherein the —N(R4)2 groups are tertiary amines.

In some aspects, the method has the proviso that the polyamine compound is not: 4 4 i R4 R4 )2 ITI/Me Mew?l 1 0 Me Me IB-2 wherein the —R4 groups are hydrogen or methyl.

In some aspects, the method has the o that the polyamine compound is not of formula IC: RZ-HN-(CH2)p-NH-(CH2)q-NHCH2-Y-CHzNH-(CH2)s-NH-(CH2)t-NH-RZ (IC) or a salt thereof; wherein Y is selected from anthracenyl, yl, and phenyl; wherein each RZ is independently selected from hydrogen and alkyl; and wherein p, q, t, and s are each an integer independently selected from 3 and 4. In some further aspects, this proviso is combined with one or more of the prior provisos.

In some aspects, the polyamine nd is selected from Ra R8 2\A3 2‘\A3 L1 A6 , ,and a salt thereof.

In some aspects, each Ra is an independently selected . In some aspects, all the Ra are the same group.

In some aspects, each A11 member is independently selected from CRa and CR5.

In some aspects, exactly one AI1 member is CRa. atively, exactly two AI1 members are each an independently selected CRa. In some aspects, at most two AI1 members are each an independently selected CRa. Alternatively, exactly three All members are each an independently selected CRa. In some aspects, the AII members are all the same CRa.

In some s, each R13, Rlb, R“, and R1d is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl. In some aspects, each R”, Rlb, R”, and R1d is a member ndently selected from hydrogen and alkyl. In some aspects, each R”, Rlb, RIC, and R1d is hydrogen.

In some aspects, each R23, R2b, RZC, R”, R2 and R2f is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, aryl, arylalkyl, or arylalkyl. In some aspects, each R”, R21), RZC, RZd, R26, and R2f is a member independently ed from hydrogen, alkyl, and fluoroalkyl. In some aspects, each Rza, R21), RZC, RZd, R23, and R2f is hydrogen.

In some aspects, each m is an integer independently selected from 1 to 16 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,10,11,12,13, 14,15, or 16). In some aspects, each m is an integer ndently selected from 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12). In some aspects, at least one m is an integer independently selected from 6 to 20 (e.g., 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20). In some aspects, at least one m is an integer independently selected from 10 to 20 (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20). In some aspects, each m is 1. In some aspects, at least one m is 1. In some aspects, each m is 2.

In some aspects, at least one m is 2.

[0253] In some aspects, each L1 and L2 is a member independently ed from a bond, -O-, and -NR4-. In some aspects, each L1 and L2 is a member independently selected from a bond and -O-. In some aspects, each L1 and L2 is a bond.

In some aspects, each R3 is a member independently selected from -Zl-R4 and -Zl-Y1- R4. In some aspects, each R3 is an independently selected -Z1-Y1-R4. In some s, each R3 is an independently selected -Z1-R4.

In some aspects, each R4 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, kyl, cycloalkylalkyl, and heteroarylalkyl. In some aspects, each R4 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, arylalkyl, and cycloalkylalkyl. In some aspects, each R4 is a member independently selected from hydrogen, alkyl, arylalkyl, and lkylalkyl. In some aspects, R4 is hydrogen, butyl, isobutyl, hexyl, or octyl.

[0256] In some s, at least one R4 is a member independently selected from alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, arylalkyl, cycloalkylalkyl, and heteroarylalkyl. In some aspects, at least one R4 is a member independently selected from alkyl, arylalkyl, and cycloalkylalkyl. In some aspects, at least one R4 is an independently selected alkyl. In some s, at least one R4 is not hydrogen. In some aspects, at least one R4 is butyl, isobutyl, hexyl, or octyl.

In some aspects, at least one pair of R4 are both a member independently selected from alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, lkyl, heteroaryl, kyl, lkylalkyl, and heteroarylalkyl. In some aspects, at least one pair of R4 are both a member independently selected from alkyl, arylalkyl, and cycloalkylalkyl. In some aspects, at least one pair of R4 are both an alkyl. In some aspects, at least one pair of R4 is not hydrogen. In some aspects, at least one pair of R4 is butyl, isobutyl, hexyl, or octyl.

In some aspects, each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, lkoxy, alkylamino, alkylaminoalkoxy, aryl, aryloxy, cycloalkyl, cycloalkoxy, cycloalkylalkoxy, halo, fluoroalkyl, fluoroalkyloxy, heteroaryl, arylalkyl, arylalkyloxy, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl. In some aspects, each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, aminoalkoxy, alkylaminoalkoxy, aryl, y, cycloalkylalkoxy, halo, lkyl, fluoroalkyloxy, arylalkyloxy, and hydroxyalkyl. In some aspects, each R5 is a member independently selected from hydrogen, alkyl, hydroxyl, , aryl, aryloxy, halo, fluoroalkyl, and fluoroalkyloxy. In some aspects, R5 is hydrogen.

In some aspects, at least one R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, lkoxy, alkylamino, alkylaminoalkoxy, aryl, y, cycloalkyl, cycloalkoxy, lkylalkoxy, halo, lkyl, fluoroalkyloxy, heteroaryl, arylalkyl, arylalkyloxy, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl. In some aspects, at least one R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, aminoalkoxy, alkylaminoalkoxy, aryl, y, cycloalkylalkoxy, halo, fluoroalkyl, alkyloxy, arylalkyloxy, and hydroxyalkyl. In some aspects, at least one R5 is a member independently selected from hydrogen, alkyl, hydroxyl, alkoxy, aryl, aryloxy, halo, fluoroalkyl, and fluoroalkyloxy.

[0260] In some aspects, each Z1 and Z2 is an independently selected -N(R4)-.

In some aspects, each R621, R613, and R60 is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, aryl, cycloalkyl, arylalkyl, heteroarylalkyl, and cycloalkylalkyl. In some aspects, each R63, R61), and R60 is a member independently selected from hydrogen, alkyl, alkyl, alkenyl, alkynyl, aryl, and arylalkyl. In some aspects, each R6a, R61), and R6C is a member independently selected from hydrogen and alkyl.

In some s, the polyamine compound comprises at least four primary or secondary amino groups. In some aspects, the polyamine compound comprises at least six primary or secondary amino groups.

[0263] In some aspects, each R81 is independently a group of Formula II: each of the An members is independently selected from CR3 and CR5; or, alternatively, a pair of adjacent AI1 members join to form an independently selected cycloalkyl, aryl, heterocyclyl, or heterocycloaryl ring that is fused with an A11 ring at the pair’s A11 ring positions; wherein at least one A11 member and at most three AI1 members are ndently selected CRa, each R”, Rlb, R”, and R1d is a member independently selected from hydrogen, fluoro, alkyl, and fluoroalkyl; for each Ra member, at most two R”1 of the Ra member are selected from - C(R2a)=(R2b)-, -cc-, and -C(Rza)(R2b)-L2-C(R2c)(R2d)-; and each m is an integer independently selected from 1 to 16.

In some aspects, the ine compound is selected from: Ra R3 fil J§A5 fi1J§A4 A2 l4 l2 A fiS’A\\AB9iv \A3. \A3 L1 AG' and a salt thereof; wherein: at least one An member and at most three A11 members are ndently selected CRa, each Rza, R21), R”, R”, R23, and R2f is a member independently ed from hydrogen, alkyl, fluoroalkyl, alkenyl, l, aryl, heteroaryl, arylalkyl, and arylalkyl; alternatively, a pair of R211 members from the Ra group independently selected from the group R2a and R21), R20 and R”, and R2e and R2f join to form a ring independently selected from spirocycloalkyl and spiroheterocycyl; or, alternatively, the R221 and the R20 from the R3 group join to form a ring independently selected from cycloalkyl and heterocycyl; each m is an integer independently selected from 1 to 12; each R3 is a member independently selected from -Z-Y1-R4 and -Z-Y1-Y2-R4; and each Z1 and Z2 is an ndently selected NR4.

[0265] In some aspects, A1, A2, A3, A4, A5, A6, A7, A8, and A9 are each an AI1 member independently selected from CRa, and CR5; or, alternatively, a pair of nt AIl members join to form a cycloalkyl, aryl, heterocyclyl, or heterocycloaryl ring; each Rza, R2b, RZC, RM, R26, and R2f is a member independently selected from hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl; each m is an integer independently selected from 1 to 10; each L1 and L2 is a member independently selected from a bond, -O-, and -NR4-; each R3 is a member independently selected from -Z-R4, -Z-Y1-R4, and —Z-Y1-Y2-R4; and each Z1 and Z2 is an independently selected NR4.

[0266] In some aspects, the invention provides the ine compound selected from: Ra R8 fi/K’F P11J§A4 5’A§Ae9 ' ' A\ 3—.A4 ‘l 2 A2 ,r —.A7 A \A3 L1 A6 and a salt thereof; , , wherein: each R"1 is independently a group of Formula V: N R20 R“: ( Rm%R2d m R3 each of the An members is independently selected from CRa and CR5; or, alternatively, a pair of adjacent AIl members join to form a cycloalkyl, aryl, heterocyclyl, or heterocycloaryl ring; wherein at least one A11 member and at most three A11 members are independently selected CRa, each Rla, Rlb, R10, and R1d is a member independently ed from hydrogen, fluoro, alkyl, and fluoroalkyl; for each Ra , at most two R”1 of the Ra member are selected from — C(Rza)=(R2b)-, —cc—, and -C(Rza)(R2b)-L2-C(R2°)(R2d)-; and each R3 is a member independently selected from R4 and -Z-Y1-Y2-R4.

[0267] In some aspects, the polyamine nd comprises at least four primary or secondary amino groups. In some aspects, the polyamine compound ses at least six primary or secondary amino .

In some aspects, the invention provides the polyamine compound selected from Ra Ra 1km 1kg) figs/L J1 A Ra, Ra A Ra, and a salt thereof; wherein each A11 member is an independently selected CR5.

In some aspects, the invention provides the polyamine compound selected from Ra Ra R5 and a salt thereof.

In some aspects, Ra is an independently selected group of Formula VII: / _\ H ( Rm)-\ m R3 VII.

In some further s, the polyamine compound is ed from Ra Ra R5 and a salt thereof.

In some aspects, R5 is hydroxyl, alkoxy, cycloalkoxy, or arylalkoxy.

In some aspects, Ra is —CH2[NH(CH2)n]pNH2; each n is an integer ndently selected from 3 to 12; and each p is an integer independently selected from 1 to 3.

In some aspects, Rm is —CH2-. In some s, m is 1. In some aspects, R5 is selected from hydrogen, phenyl, and phenyloxy.

In some aspects, the polyamine compound is selected from Ra Ra Ra and a salt thereof.

In some aspects, the polyamine compound is selected from: H2N[(H2C)3HN]2H2C\©/CH2[NH(CH2)3]2NH2 H2'\'[(H2C3)3HNl2HzC CHleH(CH2)3]2NH2 Ph H2N[(H2C3)3HN]2|‘|2C CH2[NH(CH2)3]2NH2 H2N[(HzC)3HN]2HzC CHleH(CH2)3]2NH2 CHleH(CH2)3]2NH2 and a salt thereof.

In some s, the polyamine compound is selected from: H2N[(H20)3HN]2H20 CHleH(CH2)3]2NH2 H2N[(H20)3HN]2H2C CHleH(CH2)3]2NH2 OPh and a salt f.

In some aspects, the polyamine nd is selected from Ra Ra Ra R8 AN. ”A18 kA. 5J\A31J%A4 Re SA9 RaiaM A qu\,laiiL1A 5 & L, WkiiL1AARa and a salt thereof; wherein each Ar1 member is an independently selected CR5.

[0278] In some aspects, the polyamine compound is selected from L1 R8 L1 L1 Ra L1 R5 URE UP? ; :RS and a salt thereof.

In some aspects, Ra is an independently selected group of Formula V11: V11.

In some aspects, R5 is hydrogen. In some aspects, L1 is selected from a bond and O.

In some aspects, Rm is —CH2-. In some aspects, n1 is 1.

In some aspects, the polyamine compound is selected from )3HN]2H20QCHleH(CH2)3]2NH2 HzN[(HzC)3HleHzCQCHleH(CH2)3]2NH2 )3HN]2H20 CH2[NH(CH2)3]2NH2 CH2[NH(CH2)3]2NH2, H2N[(H20)3HN]2HZCMMCHleH(CH2)3]2NH2 CHleH(CH2)3]2NH2 l‘lzl‘l[(HzC)3l‘lN12H2C O CHleH(CH2)3]2NH2 CH2[NH(CH2)3]2NH2 OiPr H2N[(H20)3HN]2H2C O CHleH(CH2)3]2NH2 H2N[(H20)3HN]2H2C CH2[NH(CH2)3]2NH2, and a salt f.

In some aspects, Ra is —CH2[NH(CH2)n]pNH2; each n is an integer independently selected from 3 to 12; and each p is an integer ndently selected from 1 to 3. In some aspects, n is 3. In some s, n is 4. In some s, n is 5, 6, 7, 8, 9, 10, 11, or 12.

In some aspects, the polyamine compound is H H N N R‘MNJ? WVNHmNHR4 or a salt thereof.

In some aspects, the polyamine compound is H H R4HNJ9 7\/NH NHR4 HN O O or a salt thereof.

[0285] In some aspects, R4 is alkyl, cycloalkyl, or arylal 1. In some asPects, R4 is alkyl.

In some aspects, R4 is isobutyl.

In some embodiments, the ine compound or combination of a polyamine compound and at least one other composition may be used to treat Gram negative and Gram positive bacteria (including strains that are resistant to conventional antibiotics), mycobacteria (including Mycobacterium tuberculosis), enveloped viruses, fungi and even transformed or cancerous cells.

In some ments, the ine compounds of the t invention are used for the treatment of cancer. The method comprises administering to a subject a composition sing a ine compound as described herein in an amount effective to reduce cancer cell proliferation or differentiation. In some embodiments, the cancer is breast cancer, a leukemia, or a melanoma. In some embodiments, the method includes administering to the subject an effective amount of a composition sing a polyamine compound as described herein and a chemotherapeutic agent. The polyamine compounds and compositions can be administered according to known methods. Such methods are described, for example, in International Patent Application No. PCT/U820 1 3/03 1 166, Which is incorporated herein by reference in its entirety.

The compounds, compositions, and s described herein can be used to kill, disperse, treat, reduce biofilms, or t or inhibit biofilm ion. In exemplary methods, the biofilms are formed by biofilm-forming bacteria. The bacteria can be a gram- ve bacterial species or a gram-positive bacterial species. Nonlimiting examples of such bacteria include a member of the genus Actinobacillus (such as Actinobacillus actinomycetemcomitans), a member of the genus Acinetobacter (such as Acinetobacter baumannii), a member of the genus Aeromonas, a member of the genus Bordetella (such as Bordetella pertussis, ella bronchiseptica, or Bordetella parapertussis), a member of the genus Brevibacillus, a member of the genus Brucella, a member of the genus Bacteroides (such as Bacteroidesfragilis), a member of the genus Burkholderia (such as Burkholderia cepacia or Burkholderia pseudomallei), a member of the genus B0relia (such as Borelia rferi), a member of the genus Bacillus (such as Bacillus anthracis or Bacillus subtilis), a member of the genus Campylobacter (such as Campylobacterjejuni), a member of the genus Capnocytophaga, a member of the genus Cardiobacterium (such as Cardiobacterium hominis), a member of the genus Citrobacter, a member of the genus Clostridium (such as idium tetani or Clostridium dijficile), a member of the genus Chlamydia (such as Chlamydia trachomatis, Chlamydia pneumoniae, or Chlamydia psiffaci), a member of the genus Eikenella (such as Eikenella corrodens), a member of the genus Enterobacter, a member of the genus Escherichia (such as Escherichia coli), a member of the genus Francisella (such as Francisella tularensis), a member of the genus Fusobacterium, a member of the genus Flavobacterium, a member of the genus Haemophilus (such as Haemophilus ducreyi or Haemophilus zae), a member of the genus Helicobacter (such as Helicobacter pylori), a member of the genus Kingella (such as Kingella kingae), a member ofthe genus Klebsiella (such as Klebsiella pneumoniae), a member of the genus Legionella (such as Legionella pneumophila), a member of the genus Listeria (such as Listeria togenes), a member of the genus Leptospirae, a member of the genus Moraxella (such as Moraxella catarrhalis), a member of the genus Morganella, a member of the genus Mycoplasma (such as Mycoplasma hominis or asma pneumoniae), a member of the genus Mycobacterium (such as cterium tuberculosis or Mycobacterium leprae), a member ofthe genus Neisseria (such as Neisseria gonorrhoeae or Neisseria meningitidis), a member of the genus Pasteurella (such as Pasteurella multocida), a member of the genus Proteus (such as Proteus vulgaris or s mirablis), a member of the genus Prevotella, a member ofthe genus Plesiomonas (such as Plesiomonas shigelloia’es), a member of the genus Pseudomonas (such as Pseudomonas aeruginosa), a member of the genus encia, a member of the genus Rickettsia (such as Rickettsia rickettsii or Rickettsia typhi), a member of the genus Stenotrophomonas (such as rophomonas hila), a member of the genus Staphylococcus (such as Staphylococcus aureus or Staphylococcus epidermidis), a member of the genus Streptococcus (such as Streptococcus viria’ans, ococcus pyogenes (group A), Streptococcus agalactiae (group B), Streptococcus bovis, or Streptococcus pneumoniae), a member of the genus omyces (such as omyces hygroscopicus), a member of the genus Salmonella (such as Salmonella enteria’itis, Salmonella typhi, or Salmonella typhimurium), a member of the genus Serratia (such as Serratia marcescens), a member of the genus Shigella, a member of the genus Spirillum (such as Spirillum minus), a member of the genus Treponema (such as Treponema pallia’um), a member of the genus Veillonella, a member ofthe genus Vibrio (such as Vibrio cholerae, Vibrio parahaemolyticus, or Vibrio vulnificus), a member of the genus Yersinia (such as Yersinia enter ocolitica, Yersinia pestis, or Yersinia pseudotuberculosis), and a member of the genus Xanthomonas (such as Xanthomonas maltophilia).

In some embodiments, the biofilm exposed to the compounds, compositions, or methods of the present invention may comprise Gram-negative or Gram-positive bacteria. In some embodiments, the bacteria are mycobacteria.

In some aspects, the biofilm comprises an antibiotic-resistant bacterial species.

[0291] The antimicrobial compounds, compositions, and methods comprising a polyamine nd may be used to control, prevent or kill biofilms in s environments. In some ments, they may be used for treating biofilms in subjects that include human or other animals. In some embodiments, they may be used for treating biofilms in medical ations such as medical s, wound dressings, t lens, oral devices, etc. In some embodiments, they may be used for treating or preventing a biofilm-related disorder. In some embodiments, they may be used for treating biofilms in industrial applications such as oil pipelines, water pipelines, water treatment at manufacturing sites, industrial flush solution, rial wash water, industrial coatings, etc. In some embodiments, they may be used for household and hygiene ations. In some embodiments, they may be used for ltural applications, such as water remediation, crop treatment, etc. In some embodiments, they may be used for food preparation ations, such as meat sprays, fruit and vegetable sanitizers.

In some aspects, the method comprises a step of coating an object with the anti- biofilm composition. In some aspects, the method comprises a step of treating a contact lens with the anti-biofilm composition.

In some embodiments, the polyamine compound or combination of a polyamine compound and at least one other composition are directed for use in industrial applications, for example oil pipelines, water treatment, water pipelines, fracking water sanitation, milk production facility ne flush solution, oil fields, paper and pulp production, machining fluids, ship coatings, shipping, paint, il sanitizers, water filtration, ling and biocorrosion, natural gas pipeline treatment, HVAC units, etc.

[0294] In some embodiments, the polyamine compound or combination of a polyamine compound and at least one other composition are directed for use in household applications, for example, sanitizing wipes, ers, toilet bowl inserts, baby care products, toys, etc.

In some embodiments, the polyamine compound or combination of a polyamine compound and at least one other composition are directed for use in nmental applications, for example, agriculture, water remediation, water treatment, crop treatment, etc.

In some aspects, the method comprises a step of treating a pipe with the iofilm composition. In some aspects, the method comprises a step of treating a heating or cooling tower with the anti-biofilm composition.

[0297] In some embodiments, the polyamine compound or combination of a polyamine compound and at least one other composition are ed for use in food production, for example, fruit and vegetable sanitizers, water systems in food production facilities, meat sprays, cooling system sanitizers, air filtration units, feed, packaging, etc.

In some aspects, the anti-biofilm ition is a paint.

In some aspects, the method ses a step of treating a patient with a biofilm- related disorder.

Some aspects of this disclosure is directed to methods of treating a biofilm-related disorder in a subject in need thereof, the method comprising administering to the t an ive amount of a polyamine compound of the present invention.

In some embodiments, the composition is administered to a surface of the t selected from the group of dermal and mucosal surfaces and combinations thereof. In other embodiments, the surface is an oral surface, a skin surface, a urinary tract surface, a vaginal tract surface, or a lung surface.

In some embodiments, the ition is administered to the subject via subcutaneous, intra-muscular, intra-peritoneal, intravenous, oral, nasal, or topical administration, and a combination thereof.

In some aspects, a subject is treated. A subject can be a mammal including, but not limited to, a primate (e.g., a monkey, such as a cynomolgous monkey, a chimpanzee, and a human). A t can be a non-human animal such as a bird (e.g., a quail, chicken, or ), a farm animal (e.g., a cow, goat, horse, pig, or sheep), a pet (e.g., a cat, dog, or guinea pig, rat, or mouse), or laboratory animal (e.g., an animal model for a disorder). Non— limiting entative subjects can be a human infant, a pre-adolescent child, an adolescent, an adult, or a senior/elderly adult.

In some embodiments, the subject is a human.

In some instances, a subject in need of treatment can be one afflicted with one or more of the infections or disorders described herein. In some aspects, the subject is at risk of developing a biofilm on or in a biologically relevant surface, or already has developed such a biofilm. Such a subject at risk can be a candidate for ent with a polyamine compound, or ation of a polyamine compound with another compound, in order to inhibit the development or onset of a biofilm—production—related er/condition or prevent the 3O recurrence, onset, or development of one or more symptoms of a biofilm-related disorder or ion. Such a subject can be harboring an immature biofilm that is ally evident or detectable to the skilled n, but that has not yet fully formed. A subject at risk of developing a biofilm can also be one in which implantation of an indwelling device, such as a medical device, is scheduled. The risk of developing a biofilm can also be due to a propensity of ping a biofilm—related disease (such as the presence of a channel transporter mutation associated with cystic fibrosis). In such subjects, a biofilm-related disorder can be at an early stage, e.g. no bacterial infection or biofilm formation is yet detected.

In certain embodiments a biofilm—related disorder is selected from pneumonia, cystic fibrosis, otitis media, chronic obstructive pulmonary disease, and a urinary tract infection and combinations thereof. In other ments, the -related disorder is a medical device-related infection. In r embodiments, the biofilm-related disorder is a periodontal disease, such as gingivitis, periodontitis or breath malodor. In still further embodiments, the biofilm—related disorder is caused by bacteria. In some embodiments, the bacteria are Gram-negative or Gram-positive bacteria. In still other embodiments, the bacteria are of the genus Actinobacillus, Acinetobacter, Aeromonas, Bordetella, Brevibacillus, Brucella, Bacteroia'es, Burkhola'eria, Borelia, us, obacter, Capnocytophaga, Cardiobacterium, Citrobacter, Clostridium, dia, Eikenella, Enterobacter, Escherichia, Entembacter, Francisella, Fusobacterium, Flavobacterium, Haemophilus, Helicobacter, Kingella, Klebsiella, Legionella, Listeria, Leptospirae, Moraxella, Morganella, Mycoplasma, Mycobacterium, Neisseria, Pasteurella, Proteus, Prevotella, monas, Pseudomonas, Providencia, Rickettsia, Stenotrophomonas, Staphylococcus, Streptococcus, Streptomyces, Salmonella, Serratia, Shigella, Spirillum, Treponema, Veillonella, Vibrio, Yersinia, or Xanthomonas.

[0307] miting examples of -related disorders include otitis media, prostatitis, cystitis, bronchiectasis, bacterial endocarditis, osteomyelitis, dental caries, periodontal disease, infectious kidney stones, acne, Legionnaire's disease, chronic obstructive pulmonary disease (COPD), and cystic fibrosis. In one specific example, subjects with cystic fibrosis display an accumulation of biofilm in the lungs and digestive tract. ts afflicted with COPD, such as emphysema and chronic bronchitis, display a characteristic ation of the airways wherein airflow through such airways, and subsequently out of the lungs, is chronically obstructed.

Biofllm-related disorders can also encompass infections derived from implanted/inserted devices, medical device-related infections, such as ions from biliary stents, orthopedic t infections, and er-related infections (kidney, vascular, peritoneal). An infection can also originate from sites where the integrity of the skin or soft tissue has been compromised. miting examples include dermatitis, ulcers from peripheral vascular disease, a burn , and trauma. For example, a ositive bacterium, such as S. pneumoniae, can cause opportunistic ions in such tissues. The ability of S. pneumoniae to infect burn wound sites, e. g., is enhanced due to the breakdown of the skin, burn-related immune defects, and antibiotic selection.

[0309] In yet other embodiments, a biofilm-related disorder is pneumonia, cystic fibrosis, otitis media, chronic ctive pulmonary disease, or a urinary tract infection. In some embodiments, the biofllm-related disorder is a medical device-related infection.

In other aspects, this sure features compounds, compositions, or methods, such as industrial, therapeutic or pharmaceutical compositions, comprising polyamine compounds in combination with one or more additional active itions.

In some instances a polyamine nd can be administered alone or in combination with a second agent, 6.g. a biocide, an antibiotic, or an antimicrobial agent, to thereby kill, disperse, treat, reduce prevent, or inhibit bacterial biofilms. An antibiotic can be co-administered with the polyamine compound either sequentially or simultaneously.

[0312] The antibiotic can be any compound known to one of ordinary skill in the art that can inhibit the growth of, or kill, bacteria. UsefiJl, miting examples of antibiotics include lincosamides (clindomycin); chloramphenicols; tetracyclines (such as tetracycline, chlortetracycline, demeclocycline, methacycline, doxycycline, minocycline); aminoglycosides (such as gentamicin, tobramycin, netilmicin, smikacin, kanamycin, streptomycin, in); beta-lactams (such as penicillins, cephalosporins, em, aztreonam); glycopeptide antibiotics (such as vancomycin); polypeptide antibiotics (such as bacitracin); macrolides (erythromycins), amphotericins; sulfonamides (such as sulfanilamide, sulfamethoxazole, sulfacetamide, iazine, sulfisoxazole, sulfacytine, sulfadoxine, mafenide, p-aminobenzoic acid, hoprim-sulfamethoxazole); methenamin; nitrofiJrantoin; phenazopyridine; trimethoprim; rifampicins; metronidazoles; cefazolins; lincomycin; spectinomycin; mupirocins; quinolones (such as nalidixic acid, cinoxacin, norfloxacin, ciprofloxacin, perfloxacin, ofloxacin, enoxacin, fleroxacin, levofloxacin); novobiocins; polymixins; gramicidins; and antipseudomonals (such as carbenicillin, carbenicillin indanyl, ticarcillin, azlocillin, mezlocillin, piperacillin) or any salts or variants thereof. Such antibiotics are commercially available, e.g., from Daiichi Sankyo, Inc. panny, NJ), Merck (Whitehouse Station, NJ), Pfizer (New York, NY), Glaxo Smith Kline (Research Triangle Park, NC), Johnson & Johnson (New Brunswick, NJ), AstraZeneca (Wilmington, DE), Novartis (East Hanover, NJ), and Sanofi-Aventis (Bridgewater, NJ). The antibiotic used will depend on the type of ial infection.

Additional known biocides include biguanide, chlorhexidine, triclosan, chlorine dioxide, and the like.

[0314] Useful examples of antimicrobial agents include, but are not limited to, Pyrithiones, especially the zinc complex (ZPT); Octopirox®; dimethyldimethylol hydantoin nt®); chloroisothiazolinone/methylisothiazolinone (Kathon CG®); sodium sulfite; sodium bisulfite; imidazolidinyl urea (Germall llS®), diazolidinyl urea (Germaill II®); benzyl l; 2-bromonitropropane-l,3-diol (Bronopol®); formalin (formaldehyde); iodopropenyl butylcarbamate (Polyphase PI 00®); chloroacetamide; methanamine; methyldibromonitrile onitrile (1,2-dibromo—2,4-dicyanobutane or Tektamer®); glutaraldehyde; 5-bromonitro-l,3-dioxane dox®); phenethyl alcohol; 0- phenol/sodium 0-phenylphenol; sodium hydroxymethylglycinate (Suttocide A®); polymethoxy bicyclic oxazolidine pt C®); dimethoxane; thimersal; dichlorobenzyl alcohol; captan; chlorphenenesin; dichlorophene; chlorbutanol; glyceryl laurate; halogenated diphenyl ethers; 2,4,4'-trichloro-2'-hydroxy-diphenyl ether (Triclosan®. or TCS); 2,2'- dihydroxy—S,5'—dibromo-diphenyl ether; phenolic nds; phenol; 2-methylphenol; 3- methylphenol; 4-methylphenol; 4-ethylphenol; 2,4-dimethylphenol; 2,5-dimethylphenol; 3,4- dimethylphenol; 2,6-dimethylphenol; 4-n-propylphenol; 4-n-butylphenol; 4-n-amylphenol; 4- tert-amylphenol; 4—n—hexylphenol; 4—n—heptylphenol; mono- and poly—alkyl and aromatic halophenols; p-chlorophenol; methyl p-chlorophenol; ethyl p-chlorophenol; n-propyl p- chlorophenol; n-butyl p-chlorophenol; n-amyl p-chlorophenol; sec-amyl p-chlorophenol; cyclohexyl p-chlorophenol; n—heptyl p—chlorophenol; n-octyl rophenol; o-chlorophenol; methyl o-chlorophenol; ethyl o-chlorophenol; n-propyl rophenol; n—butyl o- chlorophenol; n-amyl o-chlorophenol; myl o-chlorophenol; n-hexyl o-chlorophenol; n- heptyl o-chlorophenol; o—benzyl p-chlorophenol; yl-m-methyl p-chlorophenol; o- -m,m-dimethyl-p-chlorophenol; o-phenylethyl-p-chlorophenol; ylethyl-m- methyl p-chlorophenol; 3-methyl p-chlorophenol; 3,5-dimethyl p-chlorophenol; 6-ethyl-3 - methyl p-chlorophenol; 6-n—propylmethyl-p-chlorophenol; 6-isopropylmethyl-p- chlorophenol; 2-ethyl-3,5-dimethyl p-chlorophenol; 6-sec-butylmethyl p-chlorophenol; 2- isopropyl-3, 5 -dimethyl p-chlorophenol; 6-diethylmethyl-3 -methyl p-chlorophenol; 6- isopropylethylmethyl p-chlorophenol; 2-sec-amyl-3, thyl p-chlorophenol; 2- diethylmethyl-3, 5 -dimethyl p-chlorophenol; 6-sec-octylmethyl p-chlorophenol; p-chloro- m-cresol: p-bromophenol; methyl p-bromophenol; ethyl p-bromophenol; n-propyl p- bromophenol; n-butyl ophenol; n-amyl p-bromophenol; sec-amyl p-bromophenol; n- hexyl p-bromophenol; cyclohexyl p-bromophenol; o-bromophenol; tert-amyl o-bromophenol; n-hexyl o-bromophenol; n-propyl-m,m-dimethyl-o-bromophenol; 2-phenylphenol; 4-chloro- 2-methylphenol; 4-chloromethyl ; 4-chloro-3,5-dimethyl phenol; 2,4-dichloro-3,5- dimethylphenol; 3,4,5,6-tetrabromo—2-methyl—phenol; 5-methyl—2—pentylphenol; 4-isopropyl— 3-methylphenol; p-chloro-m-xylenol (PCMX); chlorothymol; phenoxyethanol; phenoxyisopropanol; 5-chlorohydroxydiphenylmethane; resorcinol and its derivatives; resorcinol; methyl resorcinol; ethyl resorcinol; n—propyl resorcinol; n-butyl inol; n-amyl resorcinol; n-hexyl resorcinol; n-heptyl resorcinol; n-octyl resorcinol; n-nonyl resorcinol; phenyl resorcinol; benzyl resorcinol; phenylethyl resorcinol; phenylpropyl resorcinol; p- chlorobenzyl resorcinol; 5-chloro 2,4-dihydroxydiphenyl methane; 4'-chloro 2,4- dihydroxydiphenyl e; 5-bromo 2,4-dihydroxydiphenyl methane; 4'-bromo 2,4- dihydroxydiphenyl methane; bisphenolic nds; 2,2'-methylene bis-(4-chlorophenol); 2,2‘—methylene bis-(3,4,6-trichlorophenol); 2,2'-methylene bis(4-chlorobromophenol); bis(2-hydroxy-3,5-dichlorophenyl)sulfide; bis(2-hydroxychlorobenzyl)sulfide; benzoic esters (parabens); methylparaben; paraben; butylparaben; araben; isopropylparaben; isobutylparaben; benzylparaben; sodium paraben; sodium propylparaben; halogenated carbanilides; -trichlorocarbanilides (e.g., Triclocarban® or TCC); 3-trifluoromethyl-4,4'-dichlorocarbanilide; 3,3',4-trichlorocarbanilide; chlorohexidine and its digluconate; diacetate and dihydrochloride; undecenoic acid; thiabendazole, hexetidine; and examethylenebiguanide) hydrochloride (Cosmocil®).

In some embodiments of any methods described herein, the method r comprises administering a e. In some embodiments, the biocide is an antibiotic.

[0316] In instances Where a polyamine compound, or combination of a polyamine compound with another compound, is to be administered to a subject, the compound or composition herein can be orated into pharmaceutical itions. The ine compound, or combination of a polyamine compound with another compound, can be incorporated into pharmaceutical itions as pharmaceutically acceptable salts or derivatives. Some pharmaceutically acceptable derivatives of the polyamine compounds of the present ion may include a chemical group, which increases aqueous solubility. As used herein, a “pharmaceutically acceptable carrier” means a carrier that can be administered to a subject together with a polyamine compound, or ation of a polyamine compound with r compound, described herein, which does not destroy the pharmacological activity thereof. Pharmaceutically acceptable carriers include, for example, solvents, binders, dispersion media, coatings, preservatives, colorants, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. mentary active compounds can also be orated into the itions.

Non-limiting examples ofpharmaceutically acceptable carriers that can be used include poly(ethylene-co-Vinyl acetate), PVA, partially hydrolyzed poly(ethylene-co-vinyl acetate), thylene-co-Vinyl acetate-co-vinyl alcohol), a cross-linked poly(ethylene-co- Vinyl acetate), a cross-linked partially hydrolyzed poly(ethylene-co-vinyl acetate), a cross- linked poly(ethylene—co-vinyl acetate-co—vinyl l), poly-D,L-lactic acid, poly—L-lactic acid, polyglycolic acid, PGA, copolymers of lactic acid and glycolic acid (PLGA), prolactone, polyvalerolactone, poly (anhydrides), copolymers of polycaprolactone with hylene glycol, copolymers of polylactic acid with polyethylene , hylene glycol; and combinations and blends thereof.

[0318] Other carriers include, e. g., an aqueous gelatin, an aqueous protein, a polymeric carrier, a cross-linking agent, or a combination thereof. In other instances, the carrier is a matrix. In yet another instances, the carrier includes water, a pharmaceutically able buffer salt, a ceutically acceptable buffer solution, a pharmaceutically acceptable idant, ascorbic acid, one or more low molecular weight pharmaceutically acceptable polypeptides, a peptide comprising about 2 to about 10 amino acid residues, one or more pharmaceutically acceptable proteins, one or more pharmaceutically acceptable amino acids, an essential-to-human amino acid, one or more pharmaceutically acceptable carbohydrates, one or more pharmaceutically acceptable carbohydrate-derived materials, a ducing sugar, glucose, sucrose, sorbitol, trehalose, mannitol, maltodextrin, dextrins, cyclodextrin, a pharmaceutically acceptable chelating agent, EDTA, DTP A, a chelating agent for a divalent metal ion, a chelating agent for a trivalent metal ion, glutathione, pharmaceutically acceptable nonspecific serum albumin, or combinations thereof.

In other embodiments, the compositions can also comprise a pharmaceutically able carrier. In still other embodiments the effective amount is an amount effective to treat or prevent a biofilm-related disorder. In some embodiments, an ive amount comprises and amount ive to treat or prevent a biofilm on a surface.

In some ments, the compositions discussed herein further comprises an agent suitable for application to the surface. In other embodiments, the ition is formulated as a wash solution, a dressing, a wound gel, or a tic tissue. In further embodiments, the composition is formulated as tablets, pills, troches, capsules, aerosol spray, solutions, suspensions, gels, pastes, creams, or foams. In some embodiments, the composition is formulated for parenteral (e.g., intravenous), intradermal, subcutaneous, oral (e.g. inhalation), transdermal al), transmucosal, l, or rectal administration.

Another aspect of this disclosure is directed to biofilm resistant l devices, comprising a surface likely to contact a biological fluid and a polyamine compound. In some embodiments, the medical device further comprises a polyamine compound, or combinations of a polyamine compound and at least one other composition, that is coated on or nated into said surface.

In some embodiments, the ine compound or combination of a polyamine compound and at least one other composition is ated as a slow-release formulation.

In some embodiments, the polyamine compound or combination of a polyamine compound and at least one other composition are directed for use in medical applications, for example, active release or passive antimicrobial coatings for l devices, lavage solutions for open wounds, oral mouthwashes, aste additives, hand sanitizers, systemic prophylactic antibiotics, lock solutions for catheters, eye drop solutions for irrigation and contact lens cleaners, prophylactic dental inserts, high level disinfectants, gastrointestinal (GI) tract oral medications for the treatment of infections such as those caused by la, sporz'dz'um, Vibrio cholerae, or Clostrz'dz'um dz'jj‘icz'le, cancer treatment including multiple myeloma, osteosarcoma, lymphoma or other forms of cancer, topical ointrnents to treat dermatological complications including infection, canker sores, psoriasis, herpes, chronic wounds, diaper rash, mycosis (athletes foot), tinea unguium (toenail fungus), ulcers, or acne, etc.

In some embodiments, the base is selected from a liquid, gel, paste, or powder. In further embodiments, the composition is selected from shampoos, bath additives, hair care preparations, soaps, lotions, creams, deodorants, are preparations, cosmetic personal care preparations, intimate hygiene preparations, foot care preparations, light tive preparations, skin g preparations, insect repellants, antiperspirants, shaving preparations, hair removal preparations, nce preparations, dental care, denture care and mouth care ations and combinations thereof.

A pharmaceutical composition containing a ine compound, or combination of a polyamine compound with another compound, can be ated to be compatible with its intended route of administration as known by those of ordinary skill in the art. Nonlimiting es of routes of administration include parenteral, e. g., intravenous, intradermal, subcutaneous, oral {e.g., inhalation), transdermal (topical), transmucosal, vaginal and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl ns; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as nediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose Vials made of glass or plastic.

Pharmaceutical itions suitable for injectable use include sterile aqueous solutions (where water —soluble) or dispersions and sterile s for the extemporaneous preparation of sterile inj ectable solutions or dispersion. For intravenous administration, suitable carriers e physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, N.J.) or ate buffered saline (PBS). In all cases, the composition can be sterile and can be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as ia and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.

The proper fluidity can be maintained, for example, by the use of a g such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. It may be desirable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition.

Prolonged absorption of the injectable itions can be accomplished by including in the composition an agent that delays absorption, for e, aluminum monostearate and n (see, e. g., Remington: The Science and ce of cy, 21st edition, Lippincott Williams & Wilkins, Gennaro, ed. (2006)).

Sterile inj ectable solutions can be prepared by orating a polyamine compound, or combination of a polyamine compound with another compound, in the ed amount in an riate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are ed by incorporating an active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile inj e solutions, the methods of preparation include, without limitation, vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a usly sterile-filtered solution thereof.

Oral compositions may include an inert diluent or an edible carrier or binders. For the purpose of oral eutic stration, a polyamine, or a combination of a polyamine compound, or combination of a polyamine compound with another compound, can be incorporated with excipients and used in the form of tablets, pills, troches, or capsules, e.g., gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash. Pharmaceutically compatible binding agents, or adjuvant als can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the ing ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon e; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, polyamine compound, or combination of a polyamine compound with another compound, can be delivered in the form of an aerosol spray from pressured ner or dispenser that contains a suitable propellant, e.g. a gas such as carbon dioxide, or a zer.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be ted are used in the formulation. Such penetrants are generally known in the art, and include, but are not limited to, for example, for transmucosal administration, detergents, bile salts, and c acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds and itions are formulated into pharmaceutically able formulation embodiments, such as nts, salves, gels, or creams as generally known in the art.

For treatment of acute or chronic wounds, polyamine compound, or combination of a polyamine compound with another compound, can be formulated as a ng, a wash solution, gel, or a synthetic tissue, etc.

The pharmaceutical compositions containing a polyamine compound, or combination of a polyamine compound with another compound, can also be prepared in the form of suppositories (e.g., with conventional itory bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

Some pharmaceutical compositions containing a polyamine compound, or combination of a polyamine compound with r compound, can be prepared with a carrier that protects the polyamine compound, or combination of a polyamine compound with another nd, against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems as described, e.g., in Tan et al., Pharm. Res. 7-2308 (2007).

Additionally, biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations are apparent to those skilled in the art.

The materials can also be obtained commercially (e.g., from Alza Corp, Mountain View, Calif). Liposomal suspensions (including mes targeted to particular cells with onal antibodies to cell surface antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, e.g., as described in US. Pat. No. 4,522,811. ty and therapeutic efficacy of such compounds and compositions can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50%> of the population) and the ED 50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LDSQ/EDso.

While nds and compositions that t toxic side effects can be used, care should be taken to design a delivery system that targets active components to the site of affected tissue in order to minimize potential damage to normal cells and, thereby, reduce side effects.

The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds and compositions lies generally within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any nds or compositions used in the methods described herein, the therapeutically effective dose can be estimated lly from cell culture assays. A dose can be ated in animal models to achieve a circulating plasma concentration range that includes the ICso (i.e., the concentration of the test compound or composition that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more tely determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography. Information for preparing and testing such compositions are known in the art. See, e.g., Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins, Gennaro, ed. (2006).

A physician will appreciate that certain factors may influence the dosage required to ively treat a subject, including but not limited to the severity of the e or disorder, previous treatments, the general health or age of the subject, and other diseases present.

Moreover, treatment of a subject with a therapeutically effective amount of a polyamine compound, or combination of a polyamine nd with another compound, can include a single treatment or a series of treatments.

The compounds or pharmaceutical compositions can be included in a container, pack, or ser together with instructions for stration. A person of ry skill in the art will appreciate that the compounds or ceutical compositions described herein can be formulated as single—dose vials. ine compounds, or combination of a polyamine compound with another compound, may be suitable as antibiofilm active substances in personal care ations, for example shampoos, bath additives, hair care preparations, liquid and solid soaps (based on synthetic surfactants and salts of saturated or unsaturated fatty acids), lotions and creams, deodorants, other aqueous or alcoholic solutions, e. g. cleansing solutions for the skin, moist cleaning cloths, oils or powders.

Any suitable amount ofpolyamine can be used in the compositions and methods of the invention. In general, the polyamines are used in concentrations ranging from about 1 ppm to about 0 ppm, or higher. The concentration of a polyamine used in a composition or method of the invention can be, for example, from about 1 to about 100,000 ppm, or from about 10 to about 10,000 ppm, or from about 100 to about 1,000 ppm, or from about 1 to about 100 ppm, or from about 1,000 to about 10,000 ppm, or from about 10,000 to about 100,000 ppm. The concentration ofa polyamine can be about 1; 2; 3; 4; 5; 6; 7; 8; 9; ; 15; 20; 25; 30; 35; 40; 45; 50; 55; 60; 65; 70; 75; 80; 85; 90; 95; 100; 125; 150; 175; 200; 225; 250; 275; 300; 325; 350; 375; 400; 425; 450; 475; 500; 525; 550; 575; 600; 625; 650; 675; 700; 725; 750; 775; 800; 825; 850; 875; 900; 925; 950; 975; 1000; 1500; 2000; 2500; 3000; 3500; 4000; 4500; 5000; 5500; 6000; 6500; 7000; 7500; 8000; 8500; 9000; 9500; ,000; 12,500; 15,000; 17,500; 20,000; 22,500; 25,000; 27,500; 30,000; 32,500; 35,000; ; 40,000; 42,500; 45,000; 47,500; 50,000; 52,500; 55,000; 57,500; 60,000; ; 65,000; 67,500; 70,000; 72,500; 75,000; 77,500; 80,000; 82,500; 85,000; ; 90,000; 92,500; 95,000; 97,500; or about 100,000 ppm. Other concentrations ofpolyamines can be usefiJl in the compositions and methods of the invention, depending in part on factors including the specific polyamine used, the presence of other active agents if any, or the species of microorganisms that are ed.

[0341] There is thus disclosed compounds, compositions, or methods comprising novel polyamine compounds, or combinations of polyamine compounds with other compounds, that have antimicrobial activity and dispersing activity against a y of ial strains capable of forming biofilms, and methods of using the same.

EXAMPLES

[0342] The following examples serve to n ments of the present disclosure in more detail. These examples should not be construed as being exhaustive or exclusive as to the scope of this disclosure.

The ing material and methods were used.

Bacterial strains. A al strain of MRSA, isolated from a patient who underwent arthroscopic knee surgery and characterized by ARUP tories, Salt Lake City, UT, was used for this study in addition to monas aeruginosa ATCC 27853 and Alcanivorax ensz's ATCC 700651. Both ATCC strains were purchased as freeze-dried pellets from ATCC. P. aeruginosa was resuspended in BHI broth, grown overnight at 37° C and transferred to fresh BHI with 30% glycerol for e at -80° C. The MRSA isolate was likewise stored in BHI with 30% glycerol at -80° C. Notably, the clinical MRSA isolate was not passaged more than three times prior to or during the study. Prior to performing MIC analysis and biofilm experiments, the frozen stocks ofMRSA and P. aeruginosa were streaked onto Columbia blood agar plates and grown overnight at 37° C. A. ensz's ATCC 70065] was resuspended from a lyophilized pellet into marine broth, grown overnight at 30° C and passaged on marine agar plates prior to experimentation.

Example 1

[0345] MIC Determination. To determine the MIC of the polyamine chains alone and the synthesized compounds against MRSA and P. aeruginosa ATCC 27853, a modified protocol from the al and Laboratory Standards Institute (CLSI) guideline M26-A was used.

Consistent with this guideline, the MIC was defined as the tration of antimicrobial that was ed to reduce approximately 5 x 105 cells per mL to approximately 5 x 102 per mL in a 24-hour period. Each MIC experiment was performed n=10 times for each antimicrobial.

All data was collected using cation adjusted MHB for standardization.

As mentioned, MRSA and P. aeruginosa ATCC 27853 were grown on Columbia blood agar plates prior to experimentation. From the blood agar plates, a 0.5 McFarland standard was made of each isolate. This equated to approximately 5 x 107 cells/mL for the MRSA isolate and approximately 3 x 107 cells/mL for the P. aeruginosa ATCC 27853 isolate. Of the 0.5 and stock solution, 50 [LL were added to MHB. All of the antimicrobials were kept in PBS stock solutions at 10 mg/mL and diluted in the MHB to reach the desired concentration that would determine the MIC value. All tests were performed such that a final volume of 5 mL ofMHB were obtained. This provided a final bacterial concentration of approximately 5 x 105 cells/mL.

Each sample was incubated in glass test tubes at 37° C for 24 hours, then plated in duplicate on tryptic soy agar (TSA) using a 10-fold dilution series. TSA plates were further incubated 24 hours at 37° C. The number of colony forming units (CFU) that grew were d in order to calculate the number of bacteria that were present per mL in the original MHB solution. The concentration of compound that ed in a reduction of approximately x 105 cells/mL to imately 5 x 102 cells/mL in a r period was defined as the MIC.

For comparison to a current rd of care, the MICs of vancomycin against the MRSA isolate and tobramycin against the P. aeruginosa ATCC 27853 isolate were also determined. MIC results for various compounds are shown in MRSA and P. aeruginosa Biofilm Eradication. s ofMRSA and P. aeruginosa ATCC 27853 were grown on the surface of PEEK nes using a membrane biofilm r. Data demonstrating the design and repeatability of this reactor have been published previously. Williams et al., In Vivo Efficacy ofa Silicone - Cationic d Antimicrobial Coating to Prevent Implant-Related Infection, Biomaterials 33, 8641—8656 (2012); Williams et al., Use ofdelrin plastic in a modified CDC biofilm reactor, Res J Microbiol 6, 425-429 (2011); Williams et al, Experimental model ofbiofilm implant-related osteomyelitis to test combination biomaterials using biofilms as initial inocula, J Biomed Mat Res A 100, 1888-1900 (2012); Williams et al., A modified CDC biofilm reactor to produce mature biofilms on the surface ofPEEK membranesfor an in vivo animal model application, Curr Microbiol 62, 663 (2011). Prior to growing the biofilms, PEEK nes were first sonicated for 10 min in medical grade detergent, rinsed with running reverse osmosis water for 10 min, sonicated in reverse osmosis water for 10 min and rinsed once again using 70% ethanol.

Four guillotine—like holders were designed to hold eight PEEK nes (two per holder). All components were washed, assembled and the reactor autoclaved prior to each use. Following American Society for Testing and Materials (ASTM) standard E2562-07, the modified reactor was run under the following conditions: approximately 5 x 107 ial cells were inoculated into 500 mL of BHI in the biofilm reactor. A paddle in the base of the reactor was stirred at 130 rpm. The unit was placed on a hot plate set at 33° C for 24 hrs. A % BHI broth solution was then fiowed through the reactor at 6.94 mL/min for an additional 24 hrs.

Following the 48-hour growth period, six of the PEEK membranes were cally removed and placed into 5 mL of MHB that contained varying concentrations of the synthesized compounds. For ison, vancomycin was also tested against MRSA biofilms and tobramycin was tested against P. aeruginosa ATCC 27853 biofilms. The membranes and broth were incubated in glass test tubes at 370 C for 24 hours. The test tubes containing membranes were vortexed for 1 minute, sonicated at 42 kHz for 10 minutes and plated in duplicate on TSA using a 10-fold dilution series and incubated at 37° C for 24 hours. This process allowed for the ination ofwhat was defined as the effective biofilm eradication concentration (EBEC) of the selected compounds. In this case, the EBEC was defined as the concentration of compound required to reduce the number of cells in the biofilms from approximately 109 cells/PEEK membrane to 105 cells/membrane. A level of 109 bacteria in the biofilms represented an amount of bacteria that may be present in one gram of soil in a natural ecosystem. See e.g., Bakken, Separation and Purification of Bacteriafrom Soil, Appl Environ Microbiol 49, 1482-1487 (1985); Torsvik et al., High Diversity in DNA ofSoil Bacteria, Applied and Environmental Microbiology 56, 782-787 (1990).

[0352] Two membranes from each run of the reactor were aseptically removed and quantified to serve as positive controls of biofilm . All of the biofilm eradication studies were repeated n=5 times. EBEC s are summarized in FIGs. 12A and 12B.

Model oil pipeline ination using Alcanivorax ensis. shows an example ofmicrobial contamination in the oil and gas industry. To test the ability and efficacy of the novel synthesized compounds to disperse and kill biofilms that may exist in a variety of settings, a model oil pipeline system was prepared to grow biofilms ofA. borkumensis, a gram-negative organism that metabolizes alkane chains. In cases of oil spills, this organism may be beneficial for bioremediation, but in this instance, it was used to represent bacteria in an oil field or pipeline that may have adverse or ed effects of oil degradation.

B shows a representative bacteria biofilm, such as the biofilm shown in FIG. l4A, d ing to certain standards in the oil and gas industry. The biofilm shown in B was d with a solution comprising 0.25% glutaraldehyde. A similar entative biofilm was treated with a solution comprising 0.5% of a polyamine compound described herein, and is shown in C. shows the results of treating biofilms of Alcam'vorax ensz's grown on the surface of galvanized steel with a solution comprising 0.25% of a polyamine compound ofthe present invention.

Combination ent. Data collected with benzylnorspermidine (CZ-7; PBC-7) at 0.25% mixed with chlorhexidine gluconate at 0.25% has shown significant dispersal of a biofilm ofMRSA that consisted of 1011 cells within one hour.

Example 2 A compound was screened for its ability to create zones of inhibition on cation adjusted Mueller Hinton agar. It is believed that the compound of Example 2 may be represented by the structure disclosed in . To perform this test, lawns of methicillin— resistant Staphylococcus aureus (MRSA) or Pseudomonas aerugz'nosa ATCC 27853 were made on cation adjusted Mueller Hinton agar using standard microbiological ques.

Immediately after g bacteria on agar, a 50 [LL drop of the compound was placed onto the agar surface. The compound was tested in duplicate at 100 ug/mL, 50 ug/mL, 25 ug/mL and 10 ug/mL. All plates were incubated at 370 C for 24 hours. Results indicated that total clearing was seen against MRSA at 10 ug/mL. Partial ng was seen against P. aerugz'nosa at 25 ug/mL and total clearing at 50 ug/mL.

The second test determined the minimum inhibitory concentration (MIC) of the compound t MRSA and P. aerugz'nosa ATCC 27853. In this instance, the MIC was defined as the amount of antimicrobial required to reduce 105 cells/mL to 102 cells/mL in a 24-hour period. This definition was tent with the al and Laboratory Standards Institute (CLSI). More specifically, the microdilution method as outlined in the M26-A CLSI guideline was used. All tests were med with cation adjusted Mueller Hinton broth.

MIC data are presented in Table l.

[0359] In addition to determining the MIC, the minimum biofilm ation concentration (MBEC) of PBC-51 was also ined. The MBEC was determined using the MBEC assay, formerly known as the Calgary Biofilm Device, developed by Innovotech. In this instance, the MBEC was defined as the amount of antimicrobial required to eradicate 100% of detectable biofilm after being grown in the MBEC device. To grow the biofilms within this device, 150 uL of on containing a concentration of 105 cells/mL were inoculated into each well of a 96-well plate. A lid containing 96 pegs was placed onto the plate and incubated for 24 hours at 370 C while being shaken at 110 rpm. Biofilms developed on the surface of the polystyrene pegs within the 96-well plate. After 24 hours, each peg contained between 105 and 106 cells in well-established biofilms. The lid with pegs was transferred to a 96-well plate that contained 200 uL of varying trations of the compound and was incubated at 37° C for an additional 24 hours. The plate was then sonicated, and the biofilms quantified using a 10-fold on series to determine the MBEC. Data are presented in Table 1.

Table 1: MIC, MBEC and EBEC of PBC-51 Against MRSA and P. aeruginosa ATCC 27853. Data are presented in ug/mL and uM concentrations. ism I IC (ug/mL / uM) MBEC (ug/mL / uM) EBEC (pg/mL/ EMRSA 1/~1.88 25/<47 250/~470.88 EP. aeruginosa 6/~11.30 35/~65.92 88.33

[0360] The fourth test that was performed was the ive biofilm eradication concentration (EBEC). To determine the EBEC, biofilms ofMRSA and P. aeruginosa ATCC 27853 were grown on the surface of PEEK membranes using a previously established protocol. Briefly, 500 mL of brain heart infusion (BHI) broth were inoculated with 105 cells/mL of MRSA or P. nosa. BHI broth was placed into a membrane biofilm reactor and placed on a hot plate set at 34° C for 24 hours with a stir bar rotating at 130 rpm.

Following the initial r growth period, a 10% BHI broth solution was flowed through the reactor at a rate of 6.944 mL/min for another 24 hours. After 48 hours of growth, each PEEK membrane had biofllm grth on it. In this experiment, there were ~2 x 1011 cells of MRSA per PEEK membrane and ~2 x 1010 cells ofP. aeruginosa per PEEK membrane. The EBEC was defined as the tration of the compound that was required to eradicate all detectable amounts (100%) of bacteria within the s. Data are presented in Table 1.

In addition to determining the antimicrobial efficacy of the compound, an initial test was performed to determine its hemolytic nature. To do so, the nd was suspended in concentrations of 100 ug/mL, 50 ug/mL and 25 ug/mL. Twenty uL of each concentration was placed onto the surface of Columbia blood agar and incubated at 37° C for 24 hours.

Results indicated that none of these concentrations displayed tic activity.

Initial tests based on visual observation were performed to determine the ability of the compound to disperse biofllms. This was done by observing the biofilms ofMRSA and P. aeruginosa that had grown on the surface ofPEEK membranes after they had been placed in three separate concentrations of the compound (300 ug/mL, 200 ug/mL and 100 ug/mL).

Biofilms were observed every 30 s to see if they ted from the PEEK membrane.

Within 2 hours, biofilms began to separate from the PEEK membranes, by 4 hours approximately half of the biofilms had separated and by 24 hours, all detectable amounts of biofilm had separated from the PEEK membrane. For comparison, it is the understanding of the inventors hereof that this dispersal effect is not seen with other antimicrobials such as chlorhexidine gluconate, glutaraldehyde, or honium chloride.

The MIC, MBEC and EBEC ofvancomycin were also determined for comparison, and the results are presented in Table 2.

Table 2: MIC, MBEC and EBEC of Vancomycin t MRSA.

Organism [MIC (ug/mL / uM) MBEC (ug/mL / uM) EBEC (ug/mL / uM) MRSA [10/673 >500/>336.50 >20,000/>l3,460 Example 3 MIC Analysis

[0364] To determine the MIC of polyamine compounds, the protocol described herein was used. The MIC is defined as being the concentration of antimicrobial (in ug/mL) ed to reduce the number of bacteria in a solution from 105 colony forming units (CFU)/mL to 102 CFU/mL in a 24-hour period.

In brief, a 0.5 McFarland of each bacterial isolate was made. A 0.5 and is a measure of turbidity in a liquid sample that contains approximately 1 x 10A8 CFU/mL. The 0.5 McFarland rd was diluted in cation adjusted Mueller Hinton Broth (CAMHB), and 50 [LL of broth were added to a well of a 96-well plate. In addition, 50 uL of CAMHB that contained a desired concentration of antimicrobial were also added to the well for a final volume of 100 [LL and a final concentration of approximately 5 x 104 CFU/well (which equated to imately 5 x 105 CFU/mL). Each well contained a desired amount of polyamine compound in order to experimentally determine the MIC. Each 96-well plate was incubated at 37° C for 24 hours. The contents of each well were plated on tryptic soy agar (TSA). TSA plates were ted for 37° C for 24 hours after which the number of CFU were counted and used to calculate the CFU/mL that remained after re to varying concentrations of compound. This procedure was repeated n=8 times for each concentration of antimicrobial. The concentration of polyamine nd that reduced bacteria from 105 CFU/mL to 102 CFU/mL in 24 hours was considered the MIC.

MICs from representative polyamine compounds as well as select antibiotics are ed in Table 3A and 3B. Consistent with what was mentioned previously, these data te that with an increase in the number of polyamine chains attached to a lipophilic ne, the MIC is lowered, indicating it has greater antimicrobial ial. CZ-7 has one chain ofnorspermidine attached, CZ-25 has two chains ofnorspermidine and CZ-51 and CZ- 52 have three chains attached.

Table 3A: MICs of Pol amine Com ounds and Select Traditional Antibiotics /mL MRSA P. aeruginosa .. MRSA — baumanmz — aeru; znosa CZ-52 3 17 ~25 400 Vancomycin 10 , , Tobramycin 7* 2 , Polymyxin B , , 2 sulfate *Not yet determined **No change Table 3B: MICs of Pol amine Com ounds and Select Traditional Antibiotics /mL 11 1i fl A Compound .. MRSA baumannzz . aeru mosa baumannzz..

CZ-25 450 >5,000 >5,000 >5,000 CZ-52 300 250 >5,000 Vancomycin , , >25,000 *Not yet determined **No change To determine the MBEC of each polyamine compound, the MBEC Inoculation Tray by Innovotech, formerly known as the Calgary biofilm device, was used. Within this device, biofilms grow on the surface of polystyrene pegs, 96 of which are ed to a lid. These pegs are inserted into a flat bottom 96-well plate. In this instance, the MBEC of a molecule was defined as the concentration of compound (in ug/mL) required to reduce 105 or 106 CFU/peg (biofilm levels varied by e) to 102 CFU/peg in a 24-hour period.

Following the manufacturer's guidelines, biofilms were grown on the surface of each peg by first making a 0.5 McFarland of each isolate. The 0.5 McFarland was diluted 1:100 in CAMHB. Into each well of a flat bottom 96-well plate, 150 uL of broth were ed. The plate was shaken at 100 rpm for 24 hours (P. aeruginosa and A. baumannii) or 48 hours (MRSA). The pegs were then placed into a te flat bottom 96-well plate for 10 seconds with 200 uL of phosphate buffered saline (PBS) in each well to remove nonadherent cells. The lid was then placed into a 96-well plate that contained g concentrations of antimicrobial with 200 uL per well. The plate was incubated for 24 hours at 37° C after which time 100 uL of broth were plated on TSA. TSA plates were incubated 24 hours at 370 C and the number of CFU counted to calculate the CFU/peg. In this instance, the MBEC was defined as the concentration of antimicrobial required to reduce 105 or 106 CFU/peg to 102 CFU/peg in a 24 hour period.

[0369] MBEC data are presented in Table 4. Similar to the MIC data, the trend of sing antimicrobial activity by increasing the number ofpolyamine chains ed to a backbone was confirmed against low number s.

EBECAnalysis In addition to determining the MIC and the MBEC ofpolyamine compounds against planktonic bacteria and low number biofilms, our group wanted to determine the efficacy of polyamine compounds t high number biofilms. To do so, biofilms were grown on the surface of polyetheretherketone (PEEK) membranes using a membrane biofilm reactor. This reactor is similar to the CDC biofilm reactor, but rather than growing biofilms on coupon surfaces, the reactor was modified to hold PEEK membranes. In short, to grow biofilms within this system, 500 mL of brain heart infusion (BHI) broth were inoculated with 1 mL of a 0.5 McFarland. The reactor was placed on a hot plate set at 34° C and the ia were grown under batch conditions for 24 hours. ing this protocol, biofilms typically grow to 109 CFU/PEEK membrane. each PEEK membrane has high number biofilms A solution of 10% BHI was then flowed through the reactor at a rate of 6.94 mL/min for an additional 24 hours. PEEK membranes were then removed and placed into 2 mL of CAMHB that contained a desired concentration of polyamine compound or antibiotic. The EBEC was defined as the concentration of antimicrobial ed to reduce a biofilm from approximately 109 CFU/PEEK membrane to approximately 102 CFU/PEEK membrane in a 24-hour period.

EBEC data are presented in Table 4. One of the most striking results was the difference in efficacy between CZ-52 and ycin. At 25,000 ug/mL, vancomycin did not have the ability to reduce biofilms of MRSA by even 1 logo unit. In contrast, at 250 ug/mL CZ-52 was able to reduce MRSA biofilms by greater than 7 logo units in a 24-hour period.

Example 4 Release OfPolyamz'ne Compoundsfrom Carrier Products

[0372] To collect proof-of-concept/preliminary data that r(s) will release these products in sufficient amounts to have antimicrobial efficacy, a polytherapy formula was ed. A Vanicream—based cream from the University of Utah compounding cy was used as the formulation base, and active ingredients were added to create a cream with final concentrations of 0.25% CZ-52, 0.25% CZ-25 and 0.1% Polymyxin B sulfate. To test its efficacy, 0.5 McFarland standards of MRSA, P. aeruginosa, and A. baumannii were made, and a lawn of each isolate was spread on Columbia blood agar. Approximately 20 mg of cream was placed in the center of each lawn. The plates were incubated 24 hours at 37° C.

Zones of clearing in each experiment ed a preliminary indication that the antimicrobials will elute out of the cream in sufficient quantities to eradicate each bacterial isolate (). Notably, the cream alone (negative control) created no zones of inhibition, and polyamine nds used alone in the cream likewise created zones of clearing. In broth samples, the polytherapy composition has shown similar results by eradicating all detectable amounts of each ial isolate after they had been grown as biofilms on PEEK membranes with approximately 109 CFU/PEEK membrane. In short, these data indicate that the polytherapy antimicrobial agents will elute out of carrier product(s) and eradicate bacteria, including those in biofilms.

Biofilm Dispersion CZ—25 and CZ-52 were tested for their ability to disperse s ofMRSA and P. aeruginosa. To test for dispersion, s of each isolate were grown on the e of commercially pure titanium (Ti) coupons (l/2” diameter x l/8” height) in a CDC biofilm reactor. The growth conditions for this reactor were the same as those for the membrane biofilm reactor. After biofilms were grown for 48 hours, each Ti coupon was aseptically d and placed into 2 mL of CAMHB for 2 hours. The CAMHB had a final concentration of 0.25% (2.5 mg/mL) CZ-25 or CZ-52. Each nd was tested n=3 times.

Following the 2-hour exposure, each coupon was fixed in 0.25% glutaraldehyde for 24 hours, dehydrated using ascending concentrations of ethanol (70%, 95%, 100%) with 3x min. changes and desiccated. One side of each coupon was carbon coated and imaged using a JEOL ISM-6610 SEM to ly observe the surface of the coupons and determine the ability of polyamine compounds to disperse biofilms compared to positive controls that had no treatment.

When compared to untreated controls, data have indicated that CZ-25 and CZ-52 have the ability to disperse biofilms from the e of a material () such that a yer of cells or reduced communities remain.

The MRSA ms were grown on the surface of um (Ti) coupons using a CDC biofilm reactor. For this reactor, an initial inoculum of N5 x 10A5 cells/mL were put into 500 mL of brain heart on (BHI) broth. The reactor was placed on a hot plate set at 34 C for 24 hours and the bacteria were allowed to grow in batch culture. For an additional 24 hours, 10% BHI broth was flowed through the reactor at a rate of 6.94 mL/min. After the 48-hour growth period, the coupons were cally removed and placed into 2 mL of cation adjusted Mueller Hinton broth (CAMHB) for 2 hours. Digital images of the biofilm dispersion process were collected after 30 minutes of exposure to polyamine compounds.

After 2 hours, the same procedure as above was used to fix the biofilms and image them by SEM.

Digital images of MRSA biofilms being dispersed from the surface of the Ti coupon also help to highlight the dispersive ability of CZ-25 ().

Data has also been collected to determine the dispersive capacity amine compounds against vorax borkumensz's, an oil-eating bacterium (FIGS. 19 and 20).

For the dispersal data with the Alcanivorax borkumensis, 1/2 inch diameter galvanized steel pipe that was approximately 6 inches long was purchased. Two strips of galvanized sheet metal were cut into 1/2 cm x 2 cm strips, and two of those were placed into the pipe. The pipe was connectedto silicone tubing, and marine broth was run through the pipes with an um of 5 x 10A5 cells/mL. The broth was flowed through the pipes at a rate of 6.94 mL/min using a peristaltic pump. Biofilms were grown on the surface of the galvanized steel for 48 hours, then exposed to polyamine compounds or aldehyde for two hours. The strips of galvanized steel were then removed, fixed in 0.25% glutaraldehyde for 2 hours (those that had already been exposed to glutaraldehyde were not fixed twice), and dehydrated in ascending concentrations of l (70%, 95%, 100%) with 3x 10 minute exchanges of each. Lastly, the strips were carbon-coated and were imaged using a JEOL JSM-6610 ng on microscope (SEM) with a lanthanum hexaboride (LaB6) filament. e 6 One of the most promising s of polyamine compounds is that preliminary data suggests that these compounds address the characteristics of biofilms that have made them resistant to antibiotic treatment. Specifically, polyamine compounds have potent crobial activity against very high inocula of bacteria and kill bacteria in a rapid fashion.

Without meaning to be bound by theory, initial characterization suggests that polyamine compounds are membrane-active, meaning they are cific in nature. This nonspecific action reduces the ial for bacteria to upregulate their defense mechanisms and reduces the risk of resistance development. From the data collected to date, it does not appear that polyamine compounds are limited to eradicating bacteria that are in log-phase growth as indicated by their activity against bacteria in the biofilm phenotype. Bacteria in a biofilm have reduced metabolic activity, which is one of the primary contributing factors that allows biofilms to resist traditional antibiotic therapy. Typically, antibiotics affect bacteria in log- phase growth. Finally, ine compounds have the ability to disperse biofilms while demonstrating antimicrobial kill. By sing and killing bacteria in a biofilm, it is hypothesized that polyamine compounds interrupt water channels and the community as a whole, allowing for distribution of polyamine compounds to a greater number, if not all, of the cells within a biofilm.

A key aspect ofpolyamine compounds is the process by which they are synthesized to provide triple action against bacteria and biofilms. The ability to synthesize antimicrobial compounds with specific activity t biofilms is cant. Traditional approaches to discovering or ping antibiotics has primarily been done through cumbersome, high— throughput screening methods with limited results wherein perhaps 1 in 1,000 compounds may be of interest. Using the below method, our group was able to produce a lead product after synthesizing approximately 20 compounds.

Organisms residing in biofilms present a complex extracellular matrix of polysaccharides (exopolysaccharides) and proteins. As a result of this complex , nutrient limiting conditions may exist that alter the normal or planktonic metabolic state. As stated above, this produces to a reduced efficacy of ional antibiotic agents, rendering them up to 1,000 times less active. A hallmark of these exopolysachharides is the presentation of acidic es from repeated glucoronic acid motifs and pyruvate derived acetals. A recent study by Losick and co-workers demonstrated that the simple polyamines spermine and norspermidine were naturally occurring inhibitors of biofilm ion, endogenously produced at high concentrations (5 0-80 uM) in response to nutrient limiting conditions and waste accumulation in mature pellicles. In this study they were able to demonstrate that norspermidine could inhibit biofilm formation at 25 uM and showed that at similar concentrations it could disperse the exopolysaccharide component of the matrix but I5 not the protein ent. This does suggest, however, that these agents might be capable of disassembling established biofilms. Interestingly, spermidine was only active at much higher trations (~ 1 mM) leading them to propose a rationale for this activity in the ability of the polyamines to engage the acidic residues in the matrix at regular intervals.

Given the lack of activity for dine vs. norspermidine, it is interesting to note that many secondary metabolites containing the polyamine frameworks have been fied as potent antimicrobials against planktonic bacteria. Most y squalamine, which ns a spermidine tail, and the related natural product incorporating a spermine tail show potent antimicrobial activity against Gram-positive and Gram-negative bacteria ().

Lysine-rich cationic peptides such as magainin and pexiganin are also potent anti-microbials against a wide range of organisms and have been developed as topical crobials for the ent of diabetic foot ulcers.

Ofnote is the incorporation of hydrophobic and cationic residues that are present in these antimicrobial “kill” compounds, without adhering to the strict en bond donor- acceptor pattern of the biofilm disruptors. Thus, an l study was made to investigate the utility of combining a simplified hobic backbone with a cationic tail that might impart molecules with the ability to inhibit biofilm formation, disrupt established biofilms and kill the emerging planktonic bacteria. The study began with the synthesis of benzylic substituted polyamines (). The synthesis of opropane substituted nes is straightforward to provide CZ-4, 12,32 from the known mono-Boc protected diaminopropane and the commercially available aldehydes (benzaldehyde, isopthalaldehyde or 1,3,5- benzenetricarboxaldehyde). This step synthetic procedure ds via reductive amination and acidic removal of the Boc group. Of note is that no purification is required until a final recrystallization of the HCl salt. The norspermidine series (CZ-7,25,52) can be prepared in a similar manner from the mono-Boc protected norspermidine. Of note is that no purification is required until a final recrystallization of the HCl salt, which allows easy preparation of these compounds on at least a ~ 10 g scale.

[0386] In some cases, increasing number of polyamine chains on the hydrophobic backbone systematically increases activity. For example, with a single chain of diaminopropane added to a benzyl backbone (CZ-4, ), the minimum inhibitory concentration (MIC; as defined by the Clinical and Laboratory Standards ute (CLSI)) against MRSA is greater than 1,200 ug/mL, whereas two chains se the MIC to 300 ug/mL ) and three chains reduce it further to 45 ug/mL (CZ-32). Similarly, with a single chain of norspermidine attached to a benzyl backbone (CZ-7, ), the MIC against MRSA is greater than 1,200 ug/mL. By adding a second chain of norspermidine (CZ-25), the MIC is 45 ug/mL. With a third chain of norspermidine attached (CZ-52), the MIC becomes 3 ug/mL.

[0387] Concurrently to the other goals of this project, a focused synthetic effort will continue to exploit this trend and generate related chemotypes with increased efficacy (e. g., compounds with four polyamine ). This is expected to be accomplished, for example, via Pd(II) mediated dimerization of 5-bromoisopthalaldehyde followed by reductive ion ().

Example 7 Evaluation ofBiofilm ation and Removal Chemistries A series of experiments is ted to investigate the impact ofbiofilm ation and removal chemistries on mimetic multispecies biofilms commonly associated with anthropogenic heating and cooling water systems.

[0389] The criteria for success is to demonstrate that these chemistries: (1) Remove a multispecies biofilm from a copper, mild steel, galvanized steel, and stainless steel substrate to a greater extent than that which can be achieved by the existing water treatment tries. (2) Do not aggressively corrode the metal ates when used in an optimum concentration. (3) Avoid material ibility issues with the existing treatment chemicals used to minimize scale formation and prevent corrosion of the metal parts of the anthropogenic heating and cooling system.

Biofilm Generation on the Metal Coupons

[0390] Normally, monoculture bacteria ing to the Pseudomonas genus have been used to investigate the physical and mechanical properties of biofilm on different heat exchanger es and substrates under changing flow conditions. However, to more accurately represent conditions that would be found in the real environment, a mixture of Pseudomonasfluorescens and Pseudomonas putida together with a 0.1% mixture ofNCH 3010 Bacillus subtilis, NCH 3032 Bacillus subtilis, NCH 3016 Bacillus licheniformis, and NCH 3040 Bacillus thuringiensis all of which are naturally occurring soil bacteria is used.

The microorganisms will be provided in pellet form from Eco-Bionics, and each pellet contains a starter e and riate nutrients that produce actively growing bacteria with the addition of water.

[0391] The experimental test procedure is a modified version of the single tube method.

Biofilm are grown on the metal coupons using a m reactor designed around a continuous stir tank reactor (). Prior to operation the reaction chamber and the individual components are disassembled, soaked in a 10% bleach solution, then scrubbed in hot soapy water and rinsed in distilled water. Once cleaned, the stir tank reactor is d with 2 liters of DI water, and 20 g of the Free Flow pellets will be added to the water.

Coupons are placed onto the coupon holding rods, which is then inserted into the Free Flow pellet on. The reactor runs for 6 days with the Free Flow solution being replaced every 2 days.

Biofilm growth isvisually observed and documented photographically. To quantify the presence of viable microorganisms within the biofilm structure formed on the metal coupons (see for an example of the typical biofilm formed on the substrate) the biofilm d coupons is removed from the reactor and lightly rinsed with sterile buffer to remove any loose debris from the surface. The coupons are then transferred to a container with 40 ml of Phosphate buffer on (PBS) and vortexed for 30 sec then placed in a sonication bath (Branson model 3200) for 30 sec. After sonication, the container with the buffer and coupon is d and vortexed again for 30 sec before being returned to the sonication bath for an additional 30 sec of processing. A third and final vortexing step ensures that the d biofilm will be thoroughly removed. After processing to remove the bioflim the PBS solution will be serially diluted onto tryptic soy agar (TSA) plates which will be incubated at 30 0C for up to 36 hours before being enumerated.

The biofilm coupons are exposed to a high, medium, and low concentration of the product (based on the recommended dosing rates) for a prescribed time. After treatment, the coupons are d, and the eluant and treatment solutions are assayed to enumerate viable bacteria A standard dispersant used for biofilm removal, together with a standard biocide (oxidizing and non-oxidizing) used at the recommended label use concentrations, are used as controls.

Corrosion Testing ent solution of the same concentrations used in the ments above is prepared with standard corrosion tor products and m deposit products in regular tap water and tested in a standard corrosion test. The solution is added to a treatment tank that is stirred continually for two weeks, after which the coupons are removed and analyzed for ion using standard corrosion test methods. Corrosion rates of less than 3.0 mpy for mild steel and 0.2 mpy for copper are considered non corrosive. A standard dispersant used for biofilm removal together with a standard biocide (oxidizing and non-oxidizing) used at the recommended label use concentrations are used as controls.

Conclusions

[0395] We have been able to determine minimum s of our polyamine compounds that will have activity against biofilms of P. aeruginosa. In general, the polyamine compounds do better against Bacilli than P. aeruginosa, so what is active against P. aeruginosa is likely active against Bacilli.

These formulas are made in PBS, water or broth.

Example 8 Methodsfor MIC, MBEC andEBECAnalysis To determine the MIC of the compounds of the present invention the ing procedure was used.

A 0.5 McFarland of A. baumanniz’ is made. A 0.5 McFarland is a measure of turbidity in a liquid sample that ns approximately 7.5 x 107 CFU/mL. The 0.5 McFarland standard is diluted in CAMHB and 50 [LL of broth is added to a well of a 96-well plate. In addition, 50 [LL of CAMHB that ns a desired concentration of antimicrobial is added to the well for a final volume of 100 uL and a final concentration of approximately 5 x 104 CFU/well (which equates to approximately 5 x 105 CFU/mL). Each well contains a desired amount of CZ nd in order to experimentally determine the MIC. Each 96- well plate is incubated at 37° C for 24 hours. The contents of each well is plated on tryptic soy agar (TSA). TSA plates is incubated at 37° C for 24 hours after which the number of CFUs is d and used to calculate the CFU/mL that remain after exposure to varying concentrations of compound. This procedure is repeated n=8 times for each concentration of antimicrobial. The concentration of CZ compound that reduces bacteria from 105 CFU/mL to 102 CFU/mL in 24 hours is considered the MIC.

To ine the MBEC, the MBEC Inoculation Tray by Innovotech, formerly known as the Calgary biofilm device, is used. Within this device, biofilms grow on the surface of polystyrene pegs, 96 of which are attached to a lid. These pegs are inserted into a flat bottom l plate. In this instance, the MBEC of a molecule is defined as the concentration of compound required to reduce approximately 106 CFU/peg to imately 102 CFU/peg in a 24-hour period.

Following the manufacturer's guidelines, ms are grown on the e of each peg by first making a 0.5 McFarland ofA. baumanm’z‘. The 0.5 McFarland is diluted 1:100 in CAMHB broth. Into each well of a flat bottom 96-well plate, 150 uL of broth is pipetted.

The plate is shaken at 100 rpm for 24 hours. The pegs are placed into a separate flat bottom 96—well plate for 10 seconds with 200 uL ofphosphate buffered saline (PBS) in each well to remove nonadherent cells. The lid is placed into a 96-well plate that contains varying concentrations of antimicrobial with 200 uL of broth per well. The plate is incubated for 24 hours at 37° C after which time 100 uL of broth is plated on TSA. TSA plates is incubated 24 hours at 37° C and the number of CFU counted to calculate the CFU/peg.

In addition to determining the MIC and the MBEC of compounds against planktonic bacteria and low number biofilms, the efficacy of the compounds of the invention against high number biofilms is determined. To do so, biofilms are grown on the surface ofPEEK membranes using a membrane biofilm reactor (Williams, D. L., Haymond, B. S. & Bloebaum, R. D. Use of delrin plastic in a modified CDC biofilm reactor. Res JMicrobiol 6, 425-429 (2011); Williams, D. L., Woodbury, K. L., Haymond, B. S., Parker, A. E. & Bloebaum, R. D. A d CDC biofilm reactor to produce mature biofilms on the surface ofPEEK membranes for an in vivo animal model application. Curr Microbiol 62, 1657-1663 (2011); Williams, D. L. et al., In Vivo Efficacy of a Silicone - Cationic Steroid Antimicrobial g to Prevent Implant-Related Infection. Biomaterials 33, 656 (2012); Williams, D. L. et al., mental model ofbiofilm implant-related osteomyelitis to test combination erials using biofilms as initial a. J Biomed Mat Res A 100, 1888-1900 (2012)).

This reactor is similar to the CDC biofilm reactor, but rather than growing biofilms on coupon surfaces, the reactor was modified to hold PEEK membranes. In short, to grow biofilms within this system, 500 mL of brain heart ll’lqulOl’l (BHI) broth is inoculated with 1 mL of a 0.5 McFarland. The reactor is placed on a hot plate set at 340 C and the bacteria grown under batch conditions for 24 hours. A solution of 10% BHI will then be flowed through the r at a rate of 6.94 mL/min for an additional 24 hours. ing this protocol, biofilms typically grow to 109 CFU/PEEK membrane. PEEK membranes is removed and placed into 2 mL of CAMHB that contain a desired concentration of the test compound. The EBEC is defined as the concentration of antimicrobial required to reduce a biofilm from approximately 109 CFU/PEEK membrane to approximately 102 CFU/PEEK membrane in a 24-hour .

MIC, MBEC and EBECAnalysis To determine the MIC of the test compounds and traditional antibiotics, a protocol which defines the MIC as being the concentration of crobial required to reduce the number of bacteria in a solution from 105 colony g units (CFU)/mL to 102 CFU/mL in a 24-hour period. MICs for tests compounds as well as for select antibiotics are provided in Table 4. Consistent with what was mentioned previously, these data indicate that, with an se in the number of polyamine chains attached to a lipophilic backbone, the MIC is lowered, indicating it has greater antimicrobial potential. CZ-7 has one chain of norspermidine attached, CZ-25 has two chains of norspermidine, and CZ-52 has three chains attached ().

The trend of increasing antimicrobial activity by increasing the number of polyamine chains attached to a backbone while also modulating the hydrophobicity of that backbone shows promising activity t A. baumannii, more specifically in the case of CZ-58 (). The analysis below shows that compounds of the invention have increased efficacy relative to current ies (e. g., polymyxin B).

MBEC data were collected using the MBEC Inoculation Tray by Innovotech. The MBEC was defined in these preliminary results as the concentration of antimicrobial required to reduce 105 or 106 CFU/peg to 102 CFU/peg in a r period. MBEC data are presented in Table 4.

—MIC ug/mL (uM) MBEC ug/mL (uM) EBEC ug/mL (uM) i 4 i Compound MRSA .. MRSA .. MRSA baumannll baumannll baumannll.. >500 >500 >500 >500 >5,000 >10,000 CZ—7 (>2,259) (>2,259) (>2,259) (>2,259) 90) (>45,180) 40 150 25 450 >5,000 cz-25 ~7,500 (110) (411) (69) (1234) (13,715) 3 400 20 >500 250 ~7,000 CZ-52 (6) (800) (40) (995) (500) (~14,000) 6 30 30 >50 1,500 400 CZ-58 (13.6) (68) (68) (>113) (3,403) (908) >500 >>25,000 Vancomycin (6.9) (>345) (>> 17,225) 1 50 Polymyxin B 1,000 sulfate (0.7) (36) (722) Table 4: MICs, MBECs and EBECs of CZ nds and ional antibiotics. Data reported in ug/mL and uM concentrations.

EBEC data were collected to determine the efficacy of CZ-52, CZ-58 and traditional antibiotics against high number biofilms ning approximately 109 CFU grown on the surface of a polyetheretherketone (PEEK) membrane. Results are shown in Table 4.

EBEC data showed striking differences in efficacy between CZ-52 and vancomycin (Table 4). At 25,000 ug/mL, vancomycin did not have the ability to reduce biofilms ofMRSA by even 1 logo unit. In contrast, at 250 ug/mL CZ-52 was able to reduce MRSA biofilms by greater than 7 loglo units in a 24-hour period. gh the MIC of CZ—58 was lower against MRSA ed to A. baumannz’z’, CZ-58 had greater activity against biofilms ofA. baumanm‘i than against MRSA biofilms in the EBEC assay. The EBEC ofpolymyxin B againstA. baumanm‘z‘ was 1,000); greater than the MIC, whereas the EBEC for CZ-58 was only ~l3x greater than the MIC. gh the MIC of CZ-58 was “high” compared to MICs of traditional antibiotics, the efficacy against biofilms was promising.

Table 5 provides MIC results for a variety of compounds against MRSA, Pseudomonas aeruginosa, and Acinetobacter baumannii.

CZ MIC MIC (P. MIC (A.

No. Compound (MRSA) aeruginosa) bumannii) CZ-3 N/\/\/NH2 H + - 2 HCI CZ-4 CZ-5 CZ—6 CZ-7 CZ—8 CZ-9 CZ MIC MIC (P. MIC (A.

No. Compound (MRSA) nosa) bumannii) + ++ 14 + 21 +++ + MIC MIC (P. MIC (A. nd (MRSA) aeruginosa) bumannii) Nl/NHHN\)/Nl-l2 LALNH NHBoc ++ ++ MIC MIC (P. MIC (A.

. (MRSA) nosa) bumannii) MIC (P. MIC (A. nosa) bumannii) +++ +++ +++ +++ ++ ++ t0 +++ CZ MIC MIC (P. MIC (A.

No. nd (MRSA) aeruginosa) bumannii) kNH NHBoc +++ +++ +++ +++ NH NH; 'x decanoic acid +++ + +++ ++ t0 +++ ++ MIC (P. MIC (A. nosa) bumannii) +++ +++ +++ +++ +++ +++ ++ +++ +++ MIC MIC (P. MIC (A.

. (MRSA) nosa) bumannii) +++ +++ +++ +++ +++ MIC MIC (P. MIC (A.

. Compound (MRSA) nosa) bumannii) +++ + +++ +++ +++ ++ CZ MIC MIC (P. MIC (A.

No. nd (MRSA) aeruginosa) bumannii) +++ +++ +++ +++ +++ ++ +++ +++ +++ +++ +++ MIC MIC (P. MIC (A.

. (MRSA) nosa) bumannii) +++ +++ : : t0 : : : Free Base MIC (P. MIC (A. nosa) bumannii) +++ +++ +++ +++ +++ +++ ++ +++ . nd (MRSA) aeruginosa) bumannii) +++ +++ +++ +++ MIC (P. MIC (A. nosa) bumannii) ++ to +++ MIC (P. MIC (A. nosa) bumannii) +++ +++ +++ +++ +++ +++ ++ ++ +++ +++ MIC (P. MIC (A. nosa) bumannii) +++ ++ to +++ ++ ++ MIC (P. MIC (A. nosa) bumannii) +++ +++ +++ +++ +++ +++ +++ +++ +++ MIC (P. MIC (A. nosa) bumannii) +++ +++ +++ +++ +++ +++ ++ +++ MIC (P. MIC (A. nosa) bumannii) —— = MIC >100 uM. = 100 uM Z MIC Z 50 uM. MIC results of “>n uM” or “<n uM” will also be designated as --+ if 100 Z n 2 50. : 50 [1M > MIC. MIC results of “>n ”M” or “<n HM” will also be ated as +++ if n < 50.

Table 5: MICs of CZ compounds.

Biofilm Dispersion The compounds of the present invention such as CZ 25, CS—58 and CS-52 have excellent sal characteristics. To test for dispersion, biofilms of each isolate were grown on the surface of commercially pure titanium (Ti) s (1/2” diameter x 1/8” height) in a CDC biofilm reactor. The growth conditions for this reactor were the same as those for the membrane biofilm reactor. After biofilms were grown for 48 hours, each Ti coupon was aseptically removed and placed into 2 mL of cation adjusted Mueller Hinton broth (CAMHB) for 2 hours. The CAMHB had a final concentration of 0.25% (2.5 mg/mL) CZ-25 or CZ-52.

Each compound was tested n=3 times.

Following the 2-hour exposure, each coupon was fixed in 0.25% glutaraldehyde for 24 hours, dehydrated using ascending concentrations of ethanol (70%, 95%, 100%) with 3x min. changes and ated. One side of each coupon was carbon coated and imaged using a JEOL ISM-6610 SEM to directly observe the surface of the coupons and determine the ability of CZ compounds to disperse biofilms compared to ve controls that had no treatment. When compared to untreated controls, data have ted that CZ—25, CZ—52 and CZ-58 have the ability to disperse biofilms from the surface of a material () such that a monolayer of cells or reduced communities .

[0409] provides SEM images of Ti coupons with MRSA biofilms exposed to CZ- (left), CZ—52 (middle) or water only (right). CZ-25 and CZ—52 demonstrated the ability to disperse the majority of biofilms such that a monolayer of cells remained on the surface of the Ti. Those samples d with water only had biofilm communities that remained in all areas of the coupons.

Example 10 Preparation 0fN1,N1 V—(I, 3-phenylenebis(methylenej)bis(N3-(3-amin0pr0pyl)pr0pane-I, 3- diamz'ne), hydrachlaride salt[C0mp0und VII-2] The ine compound (VII-2) may be synthesized as shown in Scheme 7.

Norspermidine was reacted with di-t—butyl dicarbonate compound to provide N—Boc protected norspermidine using the following procedure: To a solution of norspermidine (27.2 g, 207.7 mmol, 3.0 equiv.) in THF solvent (1000 mL) at 00C was added di-t-butyl dicarbonate (18.1 g, 83.1 mmol, 1 equiv.) se over a period of two hours. During the period of addition, the solution went from clear to a cloudy white. Following on, the reaction mixture was allowed to stir at room temperature for a period of 12 hours. The THF solvent was evaporated, and the residue was dissolved in CH2C12 (250 mL) and washed with water (250 mL). The resultant aqueous layer was washed with CH2C12 (4 x 100 mL). The c layer was dried with Na2804 and evaporated to afford the Boo-protected amine as a clear viscous liquid.

Scheme 7 y{5.1qmvxxfiAVANHfi: i {gflflj‘go MeOH. 3; A ms, than 8.133%; Beer-{N’N’”‘§”‘\X‘§’ ' {’N”\§’\f\§“\f\m~;am.E 'n L' nd ‘33an Acid. MESH li«NTN\VA§\Qr\Vx\’N”r\-fi% ANWTNNHQ H H El {f H H Compound w-z Benzene-1,3-dicarboxyaldehyde was reacted with at least two equivalents of the N- Boc protected norspermidine in the presence of molecular sieve and ol solvent to provide a ponding polyamide product, which was then reduced by sodium borohydride (NaBH4) to produce the N—Boc protected polyamine compound VII-2a using ing procedure: To a stirring solution of 4 A molecular sieve (100 mg) in MeOH solvent (25 mL) was added benzene-1,3-dicarboxyaldehyde (0.78 g, 5.87 mmol, 1 equiv.) followed by the Boo-protected amine (2.71 g, 11.74 mmol, 2 equiv.). The solution was stirred for 12 hours at room temperature. The newly formed imine was quenched at room ature by the addition ofNaBH4 (0.89 g, 23.48 mmol, 4 equiv.) after which the solution was stirred for an additional one hour. The solution was filtered through a pad of celite, and the solvent evaporated in vacuo to afford brown oil. The crude mixture was dissolved in EtOAc (100 mL) and washed with 10% NaOH (100 mL): The resulting NaOH was further washed with EtOAc (50 mL x l). The c layer was dried with Na2SO4 and concentrated under reduced pressure to provide a crude Boc-protected product.

The Boc-protecting group on the terminal amine of the polyamine compound VII- Za was deprotected by subjecting compound VII-23 to acid hydrolysis to produce the polyamine compound (VII-2) using the following ure: The crude Boc-protected product was added to a stirring solution of HCl in MeOH (100 mL) and left for one hours.

The solvent was evaporated in vacuo to yield a white to off-white solid which was recrystallized from MeOH to provide the polyamine compound (VII-2) as white solid to off- white solid (30% yield). 1H NMR (500 MHZ, D20): 7.60 (s, 4H), 4.33 (s, 4H), 3.26-3.18 (m, 12H), 3.12 (t, J=8Hz, 4H), 2.21-2.08 (m, 8H). e 11 Preparation ofNI,NI '— (((1,3-phenylenebis(methylene))bis(azanediyl))bz’sQaropane-j’, I— diyl))bis(N3,N3-dz'methylpropane-I,3-dz'amz'ne), hydrochloride salt [Compound VII-2C] The ine compound (VII-2C) may be synthesized as shown in Scheme 8.

Benzene-l ,3-dicarboxyaldehyde was reacted with at least two equivalents ofN, N- dimethylnorspermidine in the presence of molecular sieve and methanol solvent to provide a corresponding polyamide product, which was then reduced by sodium borohydride (NaBH4) to produce the polyamine compound VII-2B using the ing procedure: To a stirring solution of 4 A molecular sieve (100 mg) in MeOH solvent (25 mL) was added e-1,3- dicarboxyaldehyde (0.78 g, 5.87 mmol, 1 equiv.) followed by N, N—dimethylnorspermidine (2.71 g, 11.74 mmol, 2 equiv). The solution was stirred for 12 hours at room temperature.

The newly formed imine was ed at room ature by the addition ofNaBH4 (0.89 g, 23.48 mmol, 4 ) after which the solution was stirred for an additional one hour. The solution was filtered through a pad of celite, and the solvent evaporated in vacuo to afford brown oil. The crude mixture was dissolved in EtOAc (100 mL) and washed with 10% NaOH (100 mL). The resulting NaOH solution was further washed with EtOAc (50 mL X 1).

The organic layer was dried with Na2804 and concentrated under reduced pressure to e the polyamine compound VII-ZB.

Scheme 8 {3 H Mia [“1" \lLg;\ ’9 ‘i- BRNVWN\’%’,4\MG, MGUH, 4 A ms. than NaBm MENN,J"\\VM\N.A\~,AN9+ Me: K “NAVANMN‘X H a l H H Me M£2. w Compound VME ; Acid, MesflH xx “ck f\Nx\,~v’\ N“’«V‘N A ,MES H H '3‘ Me: ‘ ' of Me Compound Vii-2C3 The crude the polyamine nd VII-ZB was added to a stirring solution of HCl in MeOH t (100 mL) and left for one hour. The solvent was evaporated in vacuo to yield a white to off-white solid, which was recrystallized from MeOH to afford the polyamine compound VII-2C at about 30% yield as white solid to off-white solid. 1H NMR (500 MHZ, D20): 7.59 (s, 4H), 4.24 (s, 4H), 3.29-3.15 (m, 16H), 2.91 (s, 12H), 2.21-2.15 (m, 8H).

Example 12 Preparation ofN-benzyl—I,12-d0decyldiamine (V-I) The polyamine nd (V-l) may be synthesized as shown in Scheme 9 from 1,12-dodecyldiamine, either via synthetic route [A] or [B].

HZNWNHZ l (800)20 H2N/\/\/\/\/\/\/NHB0c (DPhCHO,MeOH, [A] [B] BnCLEgN 4 A mol. sieves BnHN’“\¢/\\/”\V/A\v/\\//\\/NHBOC Compound V-1a l HCI, MeOH VWNHZ Compound V-1 In the synthetic route [A], N—Boc protected—1,12—dodecyldiamine was reacted with benzyl chloride in the presence of base and solvent to provide a corresponding polyamide compound V-la. In the synthetic route [B], N-Boc protected-1,12-dodecyldiamine, prepared as described in Example 10 as following: To a solution of amine (27.2 g, 207.7 mmol, 3.0 equiv.) in THF solvent (1000 mL) at 0°C was added di-tert—butyl dicarbonate (18.1 g, 83.1 mmol, 1 equiv.) se over a period of two hours. During the period of addition, the solution went from clear to a cloudy white. Following addition, the reaction mixture was allowed to stir at room temperature for a period of 12 hours. The THF solvent was evaporated, and the residue was dissolved in CH2C12 (250 mL) and washed with water (250 mL). The resultant aqueous layer was washed with CH2C12 (4 x 100 mL). The organic layer was dried with Na2804 and evaporated to afford the N—Boc protected-l,12-dodecyldiamine amine as a clear viscous .

[0417] The N-Boc ted-1,12-dodecyldiamine was reacted with benzaldehyde in the presence of molecular sieve and methanol solvent to provide a corresponding polyamide product, which was then reduced by sodium borohydride (NaBH4) to e the N—Boc protected polyamine compound V-la using the following procedure: To a stirring solution of 4 A lar sieve (100 mg) in MeOH solvent (25 mL) was added the benzaldehyde (0.78 g, 5.87 mol, 1 equiv.) ed by the N—Boc protected-1,12-dodecyldiamine (1.36 g, 5.87 mmol, 1 equiv.). The solution was stirred for 12 hours at room temperature. The newly formed imine was quenched at room temperature by the addition 4 (0.89 g, 23.48 mmol, 4 equiv.) after which the solution was d for an additional one hour. The solution was filtered through a pad of celite, and the solvent evaporated in vacuo to afford brown oil.

The crude mixture was dissolved in EtOAc (100 mL) and washed with 10% NaOH (100 mL).

The resulting NaOH solution was further washed with EtOAc (50 mL X 1). The c layer was dried with NaZSO4 and concentrated under reduced pressure to provide the N—Boc protected polyamine compound V-la.

The Boc protecting group on the terminal amine of the polyamine compound V-la was deprotected by treating compound V-la with acid to produce the polyamine nd (V-l) using following procedure: The crude N—Boc protected polyamine compound V-la was added to a stirring solution of HCl in MeOH solvent (100 mL) and left for one hour. The solvent was evaporated in vacuo to yield a white to off-white solid, which was recrystallized from MeOH to afford the polyamine compound (V-l) at about 30% yield as a white solid to off-white solid. 1H NMR (500 MHz, D20): 7.42 (s, 5H), 4.15 (s, 2H), 3.26 (p, J= 1.5 Hz, 2H), 2.96-2.89 (m, 4H), 1.63-1.56 (m, 4H), 1.28-1.21 (m, 14H).

Example 13 Preparation 0f],3,5—trz's-[N-(3-amin0pr0pyl)—methylamz'nej-benzene (VI-I) The polyamine compound (VI-1) may be synthesized as shown in Scheme 10. fiam‘pau‘nd Vida fidzéd. MeQH Compound VM 1,3-Diaminopropane was reacted with utyldicarbonate compound to provide N—Boc—1,3—diaminopropane using following procedure: To a solution of amine (27.2 g, 207.7 mmol, 3.0 equiv.) in THF solvent (1000 mL) at 0°C was added di-tert—butyl dicarbonate (18.1 g, 83.1 mol, 1 equiv.) dropwise over a period of two hours. During the period of addition, the solution went from clear to a cloudy white. ing addition, the reaction mixture was allowed to stir at room temperature for a period of 12 hours. The THF solvent was ated, and the residue was dissolved in CH2C12 (250 mL) and washed with water (250 mL). The resultant aqueous layer was backwashed with CH2C12 (4 x 100 mL). The organic layer was dried with Na2SO4 and evaporated to afford the N-Boc-1,3-diaminopropane as a clear viscous liquid.

Benzene-1,3,5-tricarboxyaldehyde was d with at least three equivalents of three equivalents of the N-Boc protected 1,3-diaminopropane in the presence of molecular sieve and ol solvent to provide a corresponding polyamide product, which was then reduced by sodium borohydride (NaBH4) to produce the N-Boc protected polyamine compound VI-la using following procedure: To a stirring on of 4 A molecular seive (100 mg) in MeOH solvent (20 mL) was added e-1,3,5-tricarboxyaldehyde (64 mg, 0.39 mmol, 1 equiv.) followed by N -Boc protected 1,3-diaminopropane (222 mg, 1.19 mmol, 3 equiv.). The solution was stirred for 12 hours at room temperature. The newly formed imine was quenched at room temperature by the addition ofNaBH4 (90 mg, 2.38 mol, 6 equiv.) after which the solution was stirred for an additional one hour. The solution was filtered through a pad of celite, and the solvent evaporated in vacuo to afford brown oil. The crude mixture was dissolved in EtOAc (100 mL) and washed with 10% NaOH (100 mL).

The resulting NaOH was further backwashed with EtOAc (50 mL x 1). The organic layer was dried with Na2804 and concentrated under reduced pressure to provide the N-Boc protected polyamine compound VI—la.

The Boc protecting group on the terminal amine of the polyamine nd VI-la was deprotected by treating nd VI-la with acid to produce the ine compound (VI-1) using ing ure: The crude polyamine compound VI—la was added to a stirring solution of HCl in MeOH solvent (100 mL) and left for one hour. The solvent was evaporated in vacuo to yield a white to off—white solid, which was recrystallized from MeOH solvent to afford the polyamine compound VI—l at about 30% yield as white solid to off- white solid.

Example 14 Preparation 0fNI,N i,NIH—(benzene—I,3,5—trz'yltrz's(methylene))trz'sW3—is0pr0pylpr0pane—I,3— diamz'ne) (HCZ) (VII-4C) The polyamine compound (VII-4C) may be synthesized as shown in Scheme 11.

G 8 HEMW G "3' ““1“ i Q HEN’NI’A"N Me M53011, 4 ix ms, than Nam, i‘iNfiNfNNf frfie’Lfie R {jigHr“MW“\Nx‘Lm”‘1’ Ema“.

Mei‘Me Compound “£48 Acid Mei:H “1.3me x ”N”WN Me Me Me Ciammund V348 Benzene-1,3,5-tricarboxyaldehyde was reacted with at least three equivalents of N,N-dimethylnorspermidine in the presence of molecular sieve and methanol solvent to provide a corresponding polyamide product, which was then reduced by sodium borohydride (NaBH4) to produce the N-Boc protected polyamine compound VII-4B using the ing procedure: To a stirring solution of 4 A molecular sieve (100 mg) in MeOH solvent (20 mL) was added benzene-1,3,5-tricarboxyaldehyde (64 mg, 0.39 mmol, 1 equiv.) followed by N, thylnorspermidine (222 mg, 1.19 mol, 3 equiv.). The solution was stirred for 12 hours at room temperature. The newly formed imine was quenched at room temperature by the on ofNaBH4 (90 mg, 2.38 mmol, 6 equiv.) after which the solution was stirred for an additional one hour. The solution was d through a pad of celite, and the solvent evaporated in vacuo to afford brown oil. The crude mixture was dissolved in EtOAc (100 mL) and washed with 10% NaOH (100 mL). The resulting NaOH was further backwashed with EtOAc (50 mL x 1). The organic layer was dried with NaZSO4 and concentrated under reduced pressure to provide the crude N—Boc protected polyamine compound VII-4B.

The polyamine compound VII-4B was added to a stirring solution of HCl in MeOH solvent (100 mL) and left for one hour. The solvent was evaporated in vacuo to afford the polyamine compound VII-4C as viscous liquid at about 30% yield. 1H NMR (500 MHz, D20): 7.68 (s, 3H), 4.35 (s, 6H), 3.41 (p, J: 6.5 Hz, 3H), 3.23 (t, J: 7.5 Hz, 6H), 3.14-3.07 (m, 6H), .11 (m, 6H), 1.30 (d, J: 6.5 Hz, 18H). e 15 Preparation 0f],3-bz's-[N—(NL3-amin0pr0pyDpropyl—3-amine)-methylamine]-benzene (VII-2) The polyamine compound (VII-2) may be synthesized as shown in Scheme 12. N- (3 -Aminopropyl)propane-l,3-diamine (i.e., norspermidine) was reacted with di-t— butyldicarbonate compound to provide a N—Boc ted norspermidine compound.

Benzene-1,3,5-tricarboxyaldehyde was reacted with at least three lents of three equivalents of the N—Boc protected norspermidine compound in the presence of molecular sieve and methanol solvent to e a corresponding polyamide product, which was then reduced by sodium borohydride CNaBH4) to e the N-Boc protected polyamine compound VII-2:1. using the following procedure: To a stirring solution of 4 A lar sieve (100 mg) in MeOH solvent (20 mL) was added e—l,3,5—tricarboxyaldehyde (64 mg, 0.39 mmol, 1 equiv.) ed by the N-Boc protected norspermidine nd (222 mg, 1.19 mmol, 3 equiv.). The solution was stirred for 12 hours at room temperature. The newly formed imine was quenched at room temperature by the addition of NaBH4 (90 mg, 2.38 mmol, 6 equiv.) after which the solution was stirred for an additional one hour. The solution was filtered h a pad of celite, and the solvent evaporated in vacuo to afford brown oil. The crude mixture was dissolved in EtOAc (100 mL) and washed with 10% NaOH (100 mL). The resulting NaOH was further backwashed with EtOAc (50 mL x l).

The organic layer was dried with NaZSO4 and concentrated under reduced pressure to provide the crude N—Boc protected polyamine compound VII-2a.

Mrs-OH, 4 fix ms, than Naam BOCHNMm. Mw‘ M A f\ .M. . . i3! 3 i“ ill gm3’\ l NHaoc ' x i‘imN‘meHaoc N H Compound Viina Acid: MeOH ’8. X ny-n r—h ‘ A" mmNHa Compound Wk? The Boc protecting group on the terminal amine of the polyamine compound VII- Za was deprotected by treating compound VII-23 with acid to produce the polyamine compound (VII-2) using following procedure: The crude ine compound VII-23 was added to a stirring solution of HCl in MeOH solvent (100 mL) and left for one hour. The t was evaporated in vacuo to yield a white to off-white solid, which was recrystallized from MeOH to afford the polyamine compound VII-2 as white solid to off-white solid at % yield.

Example 16 Preparation of],3-bis-[N—3-amin0pr0pyl]benzenediamide (VIII-3) The polyamine compound (VIII-3) may be synthesized as shown in Scheme 13.

+ HgNwNHBOC was, (3.14205 Compound Vina—33 Acid, Meflfi {:1 C) RQNMN NMNHQ H H Compound Wild Benzene-l,3-dicarbonylchloride was reacted with at least two equivalents of the N- Boc protected 1,3-diaminopropane (prepared as described in Example 4) in the presence of triethylamine (NEtg) and romethane solvent to provide a corresponding N-Boc protected polyamide compound VIII-3a. The Boc protecting group on the terminal amine of the polyamine compound VIII-3a was deprotected by treating compound VIII-3a with acid, such as acetyl de, in methanol to produce the polyamine compound 3).

Example 17 General methodfor ation ofa’iala’ehya'e aryl starting material An aldehyde precursor to the polyamine compound may be produced by the palladium coupling method of General Synthetic Scheme 14.

General Synthetic Scheme 14 O O O O O-R5 a H H H H + X-B: O-R5 Step 1 Br X X = aryl, alkyl, vinyl, heteroaryl Reagents: (a) 5:1 DME/HZO, Pd(PPh3)4, K2C03 Specific examples of this method are set forth in the following es, such as Example 68. e 18 Preparation 0fN1,N] '—(1,3—phenylenebis(methyleneflbis(N3—(3—amm0ni0pr0pyl)pr0pane—1,3— diamz'm'um) de (CZ-25) < NHZ I"'\‘_\_/NH 3—} - 6 HCI HZN HN

[0432] Step 1: Di—tert—butyl (((((1,3—phenylenebis(methylene))bz's(azanediyl))bz's(pr0pane— 3,1-diyll)l)bis(azanediyl)‘)bis(propane-3,1—diyl))dicarbamate. Isophthalaldehyde (1.07 g, 7.99 mmol) and MeOH (50 mL) were added to a bottom flask. To the solution was added tert—butyl (3-((3 -arninopropyl)arnino)propyl)carbamate (3.69 g, 16.0 mmol) and the reaction mixture was stirred for 24 h. Sodium borohydride (1.21 g, 32.0 mmol) was added portionwise and stirred for 1 h. The reaction mixture was concentrated under d pressure, and aq. NaOH (10%, 150 mL) and EtOAc (150 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (150 mL). The organic layers were combined, dried over NaZSO4, filtered, and concentrated under reduced pressure to afford the desired product as a clear oil. The intermediate was carried forward without further purification.

Step 2: N1,NI '-(I,3-Phenylenebis(methylene))bis(N3-(3-amin0pr0pyl)pr0pane—1,3- diamz'ne): To the crude di-tert-butyl (((((1,3-phenylenebis(methylene))bis(azanediyl))- bis(propane-3,1-diyl))bis(azanediyl))bis(propane-3,1-diyl))dicarbamate from Step 1 was added methanolic HCl (150 mL, 1.0 M). The reaction mixture was stirred for 2 h and concentrated under reduced pressure. The solid was collected by vacuum filtration, and it was washed with Et20 (30 mL) and hot MeOH (30 mL) to afford the desired product (1.1 g, 56%) as a white solid. 1H NMR (500 MHz, D20) 8 ppm 7.62 (s, 4H), 4.36 (s, 4H), 3.28-3.21 (m, 12H), 3.15 (t, J: 8.0 Hz, 4H), 2.23-2.11 (m, 8H). 13C NMR (125 MHz, D20) 131.4, 131.3, 131.1, 130.2, 50.8, 44.7, 44.6, 44.1, 36.5, 23.6, 22.6. LRMS [M+H]+365.3.

Example 19 Preparation ofNI-benzylbutane-1,4-a’iaminium chloride (CZ-3) N/\/\/NH2 - 2 HCI Example 19 was prepared in a similar fashion to Example 18 ) from benzaldehyde and tert—butyl (4-aminobutyl)carbamate. 1H NMR (500 MHZ, D20) 5 7.52 (s, 5H), 4.26 (s, 2H), 3.15 (t, J= 8 Hz, 2H), 3.07 (t, J: 7.5 Hz, 2H), 1.86-1.74 (m, 4H). 13C NMR (125 MHz, D20) 130.6, 129.9, 129.7, 129.2, 51.0, 46.3, 38.8, 23.9, 22.7. LRMS [M+H]+ 179.2.

Example 20 Preparation enzylpropane-I, 3-a’iamine (CZ-4) Example 20 was prepared in a similar fashion to Example 18 (CZ-25) from benzaldehyde and tert—butyl nopropyl)carbamate. 1H NMR (500 MHz, D20) 8 7.49 (quint, J: 3 Hz, 5H), 4.26 (s, 2H), 3.17 (t, J = 8 Hz, 2H), 3.07 (t, J = 8 Hz, 2H), 2.09 (quint, J = 7.5 Hz, 2H). 13C NMR (125 MHz, D20) 5 130.6, 129.9, 129.8, 129.4, 51.4, 44.1, 36.7, 23.9. LRMS [M+H]+291.3.

Example 21 Preparation ofNI-benzyla'oa'ecane-IJZ-a'iamine, hloride salt (CZ-5) (3‘4 NH2 '2 HCI

[0436] Example 21 was prepared in a similar n to Example 18 (CZ-25) from benzaldehyde and tert—butyl (12-aminododecyl)carbamate.

Example 22 Preparation ofNI-(3-(benzylamino)propyl)-N3,N3-dimethylpropane-I, 3-diamine, hydrochloride salt (CZ-6) ©/\N/\/\N/\/\N/H H | °3HCI Example 22 was prepared in a similar fashion to Example 18 (CZ-25) from benzaldehyde and Nl-(3-aminopropyl)—N3,N3-dimethylpropane-l,3-diamine. 1H NMR (500 MHz, D20) 8 7.41 (s, 5H), 4.18 (s, 2H), 3.20-3.08 (m, 8 H), 2.83 (s, 6H), 2.15-2.03 (m, 4H). 13C NMR (125 MHz, D20) 131.0, 130.5, 130.4, 129.9, 54.8, 51.8, 45.3, 45.1, 44.5, 43.5, 23.2, 21.8.

Example 23 Preparation ofNI-(3-aminopropyl)-N3-benzylpropane-I, 3-diamine, hloride salt (CZ- ©/\H/\/\H/\/\NH2 '3HCI Example 23 was prepared in a r n to Example 18 (CZ-25) from dehyde and tert-butyl (3-((3-aminopropyl)amino)propyl)carbamate. 1H NMR (500 MHz, D20) 5 7.49 (s, 5H), 4.25 (s, 2H), 3.20—3.15 (m, 6H), 3.11 (t, J = 8 Hz, 2H), 2.17—2.07 (m, 4H). 13(3 NMR (125 MHz, D20) 5 130.3, 129.8, 129.7, 129.2, 51.2, 44.6, 44.6, 43.8, 36.5, 23.7, 22.6. LRMS [M+H]+222.2.

Example 24 Preparation ofNI-benzylhexane-I, 6—diamine, hydrochloride salt (CZ-9) -2HC| Example 24 was prepared in a similar fashion to Example 18 (CZ-25) from benzaldehyde and tert-butyl (6-aminohexyl)carbamate.

Example 25 Preparation ofNI-Benzyl-NI,N12,N12-trimethyldodecane-I,I2-diamine, hydrochloride salt «24m o”.N /N\ ~2Hm Example 25 was prepared in a similar fashion to Example 18 (CZ-25) from benzaldehyde and N1,N1 ,N12-trimethy1dodecane-1, 12-diamine.

Example 26 Preparation ofNI-benzyl-N1,N12,NIZ—trimethyldodecane-I,I2-diamine, hydrochloride salt (CZ-1 1) NW”“Q ~2Hm E:j/\H e 26 was prepared in a similar fashion to e 18 (CZ-25) from benzaldehyde and dodecane-1,12-diamine. 1H NMR (500 MHZ, D20 + CDgOD) 5 7.53 (s, 10H), 4.24 (s, 4H), 3.04 (t, J = 8.5 Hz, 4H), 1.77-1.70 (m, 4H), 1.42-1.28 (m, 16 H). 13C NMR (125 MHZ, D20 + CD3OD) 6 131.0, 129.8, 129.6, 129.3, 50.9, 47.0, 29.2, 29.1, 28.7, 26.2, 25.7.

Example 27 Preparation ofNI-benzyl-N1,N12,NU—trimethyldodecane-I,12-diamine, hydrochloride salt «24» HzNM”/\©/\”MNH2 °4HC|

[0442] Example 27 was prepared in a similar fashion to Example 18 (CZ-25) from halaldehyde and tert—butyl (3 -aminopropy1)carbamate.

Example 28 Preparation ofNI-(3-methoxybenzyl)dodecane-I,12-diamine, hydrochloride salt (CZ-13) N/\/\/\/\/\/\/NHZ - 2 HCI Example 28 was prepared in a similar fashion to Example 18 (CZ-25) from 3- methoxybenzaldehyde and tert—butyl (12-aminododecyl)carbamate. 1H NMR (500 MHz, D20) 5 7.40 (t, J= 8.0 Hz, 1H), 7.06 (s, 1H), 7.05 (s, 1H), 7.03 (s, 1H), 4.16 (s, 2H), 3.82 (s, 3H), 3.00—2.93 (m, 4H). 1.66-1.58 (m, 4H), 1.32—1.22 (m, 16H). 13c NMR (125 MHz, D20) 5158.4,131.5,129.8,121.6,114.5,114.3, 54.7, 49.8, 46.1, 38.7, 27.8, 27.7, 27.4, 25.9, 24.8, 24.5.

Example 29 Preparation ofNI-(3-aminopropyl)-N3-(3-(benzylamino)propyl)propane-1,3-diamine, hydrochloride salt (CZ— l 6) ©/\u/\/\u/\/\u/\/\NH2 °4HCI Example 29 was ed in a similar fashion to Example 18 (CZ-25) from benzaldehyde and utyl (3-((3-((3-aminopropyl)amino)propyl)amino)propyl)carbamate. 1H NMR (500 MHZ, D20) 5 7.49 (s, 5H), 4.27 (s, 2H), 3.51-3.49 (m, 6H), 3.27-3.14 (m, 6H), 3.12 (t, .1: 8.0 Hz,2H),2.21-2.10(m,4H). 13c NMR (125 MHz,D20)5130.5, 130.0, 130.0, 129.5, 51.4, 45.3, 45.2, 43.9, 43.2, 36.6, 27.8, 23.9, 22.8.

Example 30 Preparation ofNI,N1V—((2,4,6—trimethyl—I,3-phenylene)bis(methylene))bis(propane-I,3- ium) de (CZ-19) HZNM” ”MNHZ Me Me - 4 HCI Example 30 was prepared in a similar fashion to Example 18 (CZ-25) from 2,4- bis(bromomethyl)- 1 ,3 ,5 thylbenzene tert-butyl (3 -aminopropyl)carbamate.

Example 31 Preparation ofNI,NI -phenylenebis(methylene))bis(dodecane-I, IZ—diamine), hydrochloride salt (CZ-21) H2N\/\/\/\/\/\/\N NWNHZ H H - 4 HCI Example 31 was prepared in a similar fashion to Example 18 (CZ-25) from isophthalaldehyde and tert—butyl (l2-aminododecyl)carbamate. 1H NMR (500 MHZ, D20) 8 7.65 (s, 3H), 4.36 (s, 4H), 3.11 (t, J: 8.0 Hz, 4H), 3.06 (t, J: 7.5 Hz, 4H), 1.77-1.68 (m, 8H), 1.47—1.33 (m, 32H). 13c NMR (125 MHz, D20) 5 130.4, 129.9, 129.7, 128.9, 49.0, 45.6, 38.2, 27.3, 27.3, 27.2, 27.1, 26.9, 26.8, 25.4, 24.4, 24.3, 24.0. LRMS [MJrH]+ 503.5.

Example 32 Preparation of(3-(((3-((3-aminopropyl)amino)propyl)amino)methyl)phenyl)methanol, hydrochloride salt (CZ-22) \N/\/\NHH ° 3 HCI KL Example 32 was prepared in a similar fashion to Example 18 ) from isophthalaldehyde and N1,N1-dimethylpropane-1,3-diamine.

Example 33 Preparation ofNI,NI -phenylenebis(methylene))bis(hexane-I, 6-diamine), hydrochloride salt (CZ-26) H2N\/\/\/\N N/\/\/\/NH2 H H ° 4 HCI

[0448] Example 33 was prepared in a similar fashion to Example 18 (CZ-25) from isophthalaldehyde and tert—butyl (6-aminohexy1)carbamate.

Example 34 ation ofdi-tert—butyl (((((I,3—phenylenebis(methylene))bis(azanediyl))bis(propane-3,I- diyl))bis(azanediyl))bis(propane-3,I—diyl))dicarbamate (CZ-27) BocHNJPNLNH NHBoc Example 34 was prepared in a similar fashion to Example Example 18 (CZ-25) from benzaldehyde and tert—butyl -aminopropyl)amino)propyl)carbamate.

Example 35 Preparation ofNI,NI 2N1 ”— (benzene-I, 3,5-triyltris(methyleneUtrisCoropane-I,3-diamine) hydrochloride salt (CZ-32) HzNM” ”MNHZ f ° 6 HCI Example 35 was prepared in a similar fashion to Example 18 (CZ-25) from benzene-1,3,5-tricarbaldehyde and tert-butyl (3—aminopropyl)carbamate. e 36 Preparation ofNI,NI 2N1 ”— (benzene-I, 3, 5-triyltri's(methylene))tris(hexane-I, ine), hydrochloride salt (CZ-33) H2N\/\/\/\N NWNHZ H H '6 HCI Example 36 was prepared in a similar n to Example 18 (CZ-25) from benzene-1,3,5-tricarbaldehyde and tert-butyl (6-aminohexyl)carbamate.

Example 37 Preparation ofNI,NI '-(’((I, 3-phenylenebis(n1ethylene))bis(azanediyl))bis(propane-3, I- diyl))bis(N3,Nj-dimethylpropane-I,3-diamine), hydrochloride salt (CZ-43) °6HC| Example 37 was prepared in a similar fashion to Example 18 ) from isophthalaldehyde and N1-(3-aminopropyl)-N3 ,N3-dimethylpropane-1,3-diamine. 1H NMR (500 MHZ, D20) 8 7.57 (s, 4H), 4.31 (s, 4H), 3.28-3.14 (m, 16H), 2.91 (s, 12H), 2.20-2.10 (m, 8H). 13c NMR (125 MHz,D20) 5 132.1, 131.9, 131.8, 130.9, 54.8, 515,453,451, 44.8, 43.5, 23.3, 21.9.

Example 38 Preparation ofNI,NI ZN] ”— (benzene-I, 3, 5-triyltris(methylene))tris(N3,N3-dimethylpropane- 1,3-diamine), hydrochloride salt (CZ-46) \NMN NMN/ | H H | NH ° 6 HCI Example 38 was ed in a similar n to Example 18 (CZ-25) from benzene-1,3,5-tricarbaldehyde and N1,N1-dimethylpropane-1,3-diamine.

Example 39 Preparation of(3, 5-bis(((3-((3-aminopropyl)amino)propyl)amino)methyl)phenyl)-methanol, hydrochloride salt (CZ-53) HZNMEME EMNMNHz

[0454] Example 3 was prepared in a similar fashion to e 18 (CZ-25) from 5- (hydroxymethyl)isophthalaldehyde and utyl (3-((3- aminopropyl)amino)propyl)carbamate. 1H NMR (500 MHZ, D20) 5 7.57 (s, 2H), 7.55 (s, 1H), 4.72 (s, 2H), 4.35 (s, 4H), 3.26-3.19 (m, 12H), 3.13 (t, J: 7.5 Hz, 4H), 2.21-2.09 (m, 8H). 13C NMR (125 MHz, D20) 5 142.8, 132.0, 130.7, 130.0, 63.2, 51.0, 44.9, 44.8, 44.4, 36.7, 23.9, 22.9.

Example 40 Preparation oftri-tert—butyl— (((((benzene-1,3, 5-triyltris lene))tris (azanediyl))tris- (propane-3, 1-diyl))tris(azanediyl))tris(propane-3, I -diyl))tricarbamate, hloride salt (CZ-54) O O J< H H H H H H °6HC| OxJ<O

[0455] e 40 was prepared in a r fashion to Example 18 (CZ-25) from benzene-1,3,5-tricarbaldehyde and tert-butyl (3—((3-aminopropyl)amin0)pr0pyl)carbamate.

Example 41 Preparation ofNI,N1'-(1,3-phenylenebis(methylene))bis(N3-(3-((3- aminopropyl)amino)propyl)propane—1,3-diamine), hydrochloride salt (CZ-57) HZNMEMEMNUNMHMEMNHZ °8HCI Example 41 was prepared in a similar fashion to Example 18 (CZ-25) from isophthalaldehyde and tert—butyl (3-((3-((3 -amin0pr0pyl)amin0)pr0pyl)amin0)pr0pyl)- carbamate.

Example 42 Preparation of3,3 '-(isophthaloylbis(azanediyl))bis(propanaminium) chloride (CZ-40) Step 1: tert—BuW/(5-(3-«3-((tert—butoxycarbonyl)amin0)pr0pyl)carbamoyl)phenyl)- -oxopentyl)carbamate. Isophthaloyl chloride (344 mg, 1.70 mmol), triethylamine (343 mg, 3.41 mmol), and CHzClz (10 mL) were added to a round-bottom flask. To the solution was added tert-butyl (3-aminopropyl)carbamate (593 mg, 3.41 mmol), and the reaction e was stirred for 16 h. The mixture was washed with aq. NaOH (10%, 50 mL) and the layers separated. The s layer was extracted with CHZCIZ (2 x 50 mL) and organic layers were combined, dried over NagSO4, filtered, and concentrated under reduced pressure. Purification by column chromatography (hexanes/EtOAc) afforded the desired product, which was used without further purification.

Step 2: 3,3 '—(Isophthaloylbis(azanediyl))bisQJropan-I-aminium) de. To the crude tert—butyl(5 -(3-((3-((tert-butoxycarbonyl)amino)propyl)carbamoyl)phenyl) oxopentyl)carbamate from Step 1 was added methanolic HCl (150 mL, 1.0 M). The reaction mixture was stirred for 2 h and concentrated under reduced pressure. The solid was collected by vacuum ion and washed with EtzO (30 mL) and hot MeOH (30 mL) to afford the desired product (0.388 g, 56%) as a white solid.

Example 43 Preparation ofNI,NI 2N1 ”—(benzene—I, iyltris(methylene))tris(N3—(3— aminopropyl)propane-I3-diamine) hydrochloride salt (CZ-52) H2N\V/J;:;]NLlfi:\lLH2 -9HCI H2 Step 1: Tri-tert—butyl (((((benzene-I,3,5-triyltris(methylene))tris(azanedin)- trisCoropane-j’, ))tris (azanedin)tris(propane-3, I—din)tricarbamate. Benzene-1,3,5- tricarbaldehyde (1.80 g, 11.04 mmol) and MeOH (50 mL) were added to a round-bottom flask. To the solution was added trimethyl orthoformate (165.6 g, 17.6 mmol) and tert-butyl (3 -((3 -aminopropyl)amino)propyl)carbamate (7.66 g, 33.1 mmol) and the reaction mixture was stirred for 24 h. Sodium borohydride (1.67 g, 44.2 mmol) was added portionwise and stirred for 1 h. The reaction mixture was trated under reduced pressure and aq. NaOH (10%, 150 mL) and EtOAc (150 mL) were added. The layers were ted and the aqueous layer was extracted with EtOAc (150 mL). The organic layers combined, dried over NaZSO4, filtered, and concentrated under reduced pressure to afford the desired product as a clear oil.

The intermediate was carried forward without further purification.

Step 2: N’,N’ 2N] zene-I,3,5-trz'yltrz's(methylene))tris(N3-(3- aminoproprpropane-1,3-a’z'amz'ne), hydrochloride salt. To the crude tri-tert-butyl (((((benzene- 1 ,3 ,5 -triyltris(methylene))tris(azanediyl))tris(propane-3 ,1- diyl))tris(azanediyl))tris(propane-3,1-diyl))tricarbamate from Step 1 was added methanolic HCl (150 mL, 1.0 M). The reaction mixture was d for 2 h and concentrated under reduced pressure. The solid was collected by vacuum filtration and washed with EtzO (30 mL) and hot MeOH (30 mL) to afford the desired product (5.6 g, 61%) as a white solid.

LRMS [M+H]+ 508.5.

Example 44 Preparation oftert—butyl (3—((3-((3,5-bis(((3-((3-aminopropyl)amino)propyl)amz'no)methyl)- )amino)propyl)amino)propyl)carbamate (CZ-51) H H HZNJg l/NH NHBOC WNH °8HCI HNWNHZ

[0461] Example 44 was prepared in a similar fashion to Example 43 (CZ—52) from benzene- tricarbaldehyde and tert-butyl (3-((3 -aminopropyl)amino)propyl)carbamate.

Example 45 Preparation ofN’,N”_([1,1Lbz‘phenyzj—3,5—diy1bis(methyzene))MSW—(3— apropyl)pr0pane-I,3-diaminz'um) chloride (CZ-58) < NH2 HN_'\_/NH 3“)”

[0462] Step 1: [1,1'-Biphenyl]-3,5-dicarbaldehyde. A on of 5- bromoisophthaldehyde (40.0 g, 187.8 mmol) (Med. Chem. Commun., 2012, 3, 763—770), phenylboronic acid (22.9 g, 187.8 mmol) and potassium carbonate (64.8 g, 469.5 mmol) in DME/HZO (5:1, 600 mL) was purged with N2 for 5 min. Tetrakispalladium nylphosphine (1.1 g, 0.9 mmol) was added, and the reaction mixture was heated to 90 °C and stirred for 16 h. The reaction mixture was cooled to rt, filtered through a pad of Celite diatomaceous earth, and the solvent evaporated under d pressure. Purification by column chromatography (10% EtOAc/hexanes) afforded the desired product (74%, 29.1 g) as an off-white solid. 1H NMR (300 MHz, CDClg) 8 ppm 10.18 (s, 2H), 8.37 (d, J: 1.2 Hz, 2H), 8.35 (t, J: 1.5 Hz, 1H), .66 (m, 2H), 7.55—7.43 (m, 3H).

[0463] Step 2: Di—tert—butyl ((((([I, I '—bz'phenyl]-3,5-dlylbis(methylene))bis(azanediy0)bis- (propane-3, I-diyl))bis(azanediywbismropane-i ))dicarbamate. [1 , 1‘-Biphenyl]—3,5- dicarbaldehyde (3.62 g, 17.24 mmol) and MeOH (100 mL) were added to a round-bottom flask. To the solution was added tert-butyl (3-((3—aminopropyl)amino)propyl)carbamate (7.96 g, 34.4 mmol), and the reaction mixture was stirred for 24 h. Sodium borohydride (2.62 g, 69.0 mmol) was added, and the mixture was stirred for 1 h. The solvent was d under reduced pressure to afford a white solid. Aqueous NaOH (10%, 200 mL) was added, and the solution was extracted with EtOAc (200 mL). The organic layer was separated, and the aqueous layer was extracted with EtOAc (200 mL). The organic layers were combined, dried over Na2S04, filtered, and concentrated under reduced pressure to afford the desired product as a clear oil that was used without further purification.

Step 3: N1,NII-([I,I '-bz'phenyl]-3,5—dl'ylbis(methylene))bis(N3-(3- ammoniopropyl)pr0pane-1,3-dz'amz'm'um) chloride. To the crude N1,N1'-([1,l'-biphenyl]-3,5- diylbis(methylene))bis(N3—(3—ammoniopropyl)propane—1,3—diaminium) chloride from Step 2 was added methanolic HCl (150 mL, 1.0 M). The reaction e was stirred for 2 h and concentrated under reduced pressure. The solid was collected by vacuum filtration and washed with EtzO (30 mL) and hot MeOH (30 mL) to afford the desired t (6.8 g, 60%) as a white solid. 1H NMR (D20, 500 MHZ) 5 ppm 7.85 (s, 2H), 7.74 (d, J: 7.5 HZ, 2H), 7.59-7.56 (m, 3H), 7.50-7.49 (m, 1H), 4.38 (s, 4H), 3.29 (t, J=8.0 Hz, 4H), 3.23 (q, J= 5.0 Hz, 8H), 3.14 (t, J: 8.0 Hz,4H),2.25-2.19(m,4H),2.17-2.11 (m, 4H). 13c NMR (125 MHz, D20) 142.3, 138.8, 132.1, 130.0, 129.5, 129.2, 128.4, 127.0, 50.8, 44.7, 44.6, 44.2, 36.5, 23.7, 22.7. LRMS [M+H]+441.4.

Example 46 Preparation 1 '—((5-(benzo[d][l,3]di0x0[—5—yl)-I,3-phenylene)bisMethylene»bis(N3- (3-amm0ni0pr0py0pr0pane-I,3-dz'amz'm'um) chloride (CZ-61) < NH2 HN—\_/NH HN OW j? 0 O 0 HZN HN -6HC| Example 46 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, benzo[d][1,3]dioxolylboronic acid, and tert—butyl (3 -((3- aminopropyl)amino)propyl)carbamate. N1,N1'-((5 -(benzo[d][1,3]dioxol-5 -yl)-1,3- phenylene)bis(methylene))bis(1\73-(3-aminopropyl)propane-1,3-diamine), hloride salt: 1H NMR (500 MHz, D20) 8 7.72 (s, 2H), 7.49 (s, 1H), 7.15 (s, 2H), 6.93 (d, J = 8.5 Hz, 1H), .95 (s, 2H), 4.30 (s, 4H), 3.21—3.12 (m, 8H), 3.05 (t, J 2 8.0 Hz, 4H), 2.17—2.02 (m, 8H). 13c NMR(125 MHz, D20)8148.1, 147.6, 142.3, 133.3, 132.2, 130.8, 129.7, 129.6, 129.4, 121.1, 109.0, 107.5, 101.6, 51.0, 44.8, 44.8, 44.4, 36.6, 23.9, 22.8.

Example 47 Preparation ofNI,N1 EN] ”-([1, I enyl]-3,3 ',5-triyltris(methylene))tris(N3-(3— aminoproprpropane-1,3-diamine), hydrochloride salt (CZ-62)

[0466] Example 47 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, (3-f0rmy1phenyl)b0r0nic acid and tert-butyl (3-((3- aminopropy1)amin0)pr0pyl)carbamate. 1H NMR (400 MHZ, D20) 5 7.96 (s, 2H), .85 (m, 2H), 7.71-7.61 (m, J= 8.5 Hz, 3H), 4.47 (s, 4H), 4.44 (s, 2H), 3.36-3.23 (m, 18H), 3.17 (t, J: 8.0 HZ, 6H), 2.30-2.22 (m, 6H), 2.20-2.13 (m, 6H). LRMS [M+H]+ 584.5.

Example 48 Preparation ofN’,N’ '-((2'-methyl-[1,1 '-biphenyl]-3, 5-diyl)bis(methylene))bis(N3—(3- aminoproprpropane-1,3-diamine), hydrochloride salt (CZ-63) HN_\_/NH Example 48 was prepared in a r fashion to Example 45 (CZ-5 8) from 5— bromoisophthaldehyde, o-tolylboronic acid and tert—butyl (3-((3- aminopr0py1)amino)propyl)carbamate. 1H NMR (500 MHZ, D20) 5 7.59 (s, 1H), 7.56(d, J = 1.0 HZ, 2H), 7.41—7.40 (m, 2H), 7.38—7.35 (m, 1H), 7.32—7.31 (m, 1H), 4.38 (s, 4H), 3.25 (t, J =7.5 Hz, 4H), 3.20-3.18 (m, 8H), 3.11 (t, J = 8.0 HZ, 4H), 2.25 (m, 3H), 2.20-2.15 (m, 4H), 2.14—2.07 (m, 4H) ). 13c NMR (125 MHZ,D2O)8143.3,139.9,135.6,131.8,131.7,131.5, 131.0, 130.5, 129.9, 129.7, 128.2, 126.1, 50.8, 44.7, 44.6, 44.2, 36.6, 23.7, 22.7. LRMS [M+H]+455.4.

Example 49 Preparation 1 '-('(4 '-morpholino-[1, 1 '-biphenyl]-3,5-diyl)bis(methylene))bis(N3- (3- aminoproprpropane-I,3-diamine), hydrochloride salt (CZ-64) MP 6km"...

Example 49 was prepared in a similar n to Example 45 (CZ-58) from 5- bromoisophthaldehyde, (4-m0rpholin0phenyl)b0r0nic acid and tert-butyl (3-((3- aminopropy1)am1n0)propyl)carbamate.

Example 50 ation ofN],NI',N1"-([1, 1'-biphenyl]-3,4C5-triyltris(methylene))tris(N3-(3— aminoproprpropane-1,3-diamine), hydrochloride salt (CZ-65) < NH2 l'm_\_/NH HZNjHM? 0 O NH -9HC| (—/ HN NH2

[0469] Example 50 was ed in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, (4-f0rmy1phenyl)boronic acid and tert-butyl (3-((3- aminopropy1)amino)propyl)carbamate. 1H NMR (500 MHZ, D20) 5 7.92 (s, 2H), 7.84 (d, J = 8.5 Hz, 2H), 7.65 (d, J = 8.5 Hz, 3H), 4.43 (s, 4H), 4.37 (s, 2H), 3.31-3.28 (m, 6H), 3.26-3.20 (m, 12H), 3.14 (t, J = 8.0 Hz, 6H),2.25—2.17 (m, 6H), 2.16-2.10(m, 6H). 13C NMR (125 MHz, D20) 8 141.6, 140.1, 132.2, 130.6, 130.4, 129.7, 127.8, 50.8, 44.7, 44.6, 44.2, 36.5, 23.7, 22.7, 22.6. LRMS [M+H]+584.5.

Example 51 Preparation ofN’,N’ 1N1 ("-(4'-l'sopropoxy-[I,I '-bz'phenyl]-3,3 ', 5-trz'yl)tris(methylene))- tris(N3-(3-aminopropyl)propane-I,3-diamine), hydrochloride salt (CZ-66)

[0470] Example 51 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- sophthaldehyde, (3-f0rmylisopropoxyphenyl)boronic acid and tert-butyl (3-((3- ammopropyl)amin0)pr0pyl)carbamate. 1H NMR (500 MHZ, D20) 5 7.93 (s, 2H), 7.88-7.82 (m, 2H), 7.65 (s, 1H), 7.35 (d, J = 9.0 Hz, 1H), 4.94-4.87 (m, 1H), 4.47 (s, 4H), 4.43 (s, 2H), 3.39-3.28 (m, 18H), 3.22 (t, J = 8.0 Hz, 6H), 2.33-2.18 (m, 12H), 1.48 (d, J = 6.5 Hz, 6H). 13C NMR (125 MHz, D20)8158.9,143.9,134.8,133.8,132.9,132.6,132.3,131.8,122.7, 117.0, 84.4, 74.5, 53.5, 49.6, 47.3, 47.3, 47.0, 46.9, 46.7, 39.2, 26.3, 25.3, 25.2, 23.9. LRMS [M+H]+642.5.

Example 52 Preparation ofNI,NI '-(’(4 '-z'sopropoxy-[I, I enyl]-3,5-dz'yl)bis(Methylene))bis(N3-(3- aminoproprpropane-1,3-dz'amz'ne), hloride salt (CZ-67) {NHNHZ HN—\_/ M)—Me H2N3—?0000 -6HC| e 52 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, (4-isopropoxyphenyl)boronic acid, and tert-butyl (3-((3- aminopropyl)amino)propyl)carbamate. 1H NMR (500 MHz, D20) 5 7.81 (s, 2H), 7.68 (d, J = 8.5 Hz, 2H), 7.54 (s, 1H), 7.12 (d, J: 8.5 Hz, 2H), 4.78-4.72 (m, 1H), 4.37 (s, 4H), 3.28 (t, J = 8.0 Hz, 4H), 3.24-3.19 (m, 8H), 3.13 (t, J= 8.0 Hz, 4H), 2.24-2.19 (m, 4H), 2.17-2.09 (m, 4H), 1.35 (d, J= 6.0 Hz, 6H). 13C NMR (125 MHz,D20)6157.0, 141.8, 132.0, 131.8, 129.4, 129.0, 128.3, 116.8, 71.6, 50.8, 44.6, 44.6, 44.1, 36.4, 23.6, 22.6, 21.0. e 53 Preparation I '-((5-(thz'0phenyl)-1,3-phenylene)bis(methylene))bisfl\73-(3- aminopropyl)pr0pane-1,3-a’z'amz'ne), hydrochlarz'de salt ) < NH2 l"N_\_/NH )7HN S HZN HN '6HCI Example 53 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, (4-(thi0phenyl)phenyl)boronic acid and tert-butyl (3-((3- aminopropyl)amino)propyl)carbamate. 1H NMR (500 MHz, D20) 8 7.90 (s, 2H), 7.83 (t, J = 1.0 Hz, 1H), 7.63-7.61 (m, 1H), 7.60-7.57 (m, 1H), 7.53 (s, 1H), 4.38 (s, 4H), 3.27 (t, J = 8.0 Hz, 4H), 3.23-3.19 (m, 8H), 3.13 (t, J = 8.0 Hz, 4H), 2.23-2.17 (m, 4H), 2.15-2.09 (m, 4H). 13c NMR(125 MHz,D20)8139.9, 137.1, 1322,1296, 128.7, 127.7, 125.9, 122.3, 50.8, 44.7, 44.6, 44.1, 36.5, 23.7, 22.6. LRMS [M+H]+ 447.3.

Example 54 Preparation ofN’,N’ 2N1"—((4Larmuoromethyu—[L1 '-biphenyl]-3,3',5- triyl)tris(methylene))tris(N3-(3-aminopropyl)propane-I,3-dz'aml'ne), hydrochloride salt (CZ- -9HC| g Example 54 was prepared in a similar fashion to e 45 (CZ-5 8) from 5- bromoisophthaldehyde, (3-formyl(trifluoromethy1)phenyl)boronic acid and tert-butyl (3- ((3-aminopropyl)amino)propyl)carbamate. 1H NMR (500 MHZ, D20) 8 7.93 (s, 2H), 7.87 (s, 2H), 7.61 (s, 2H), 4.45 (s, 2H), 4.38 (s, 4H), 3.27-3.14 (m, 16H), 3.10-06 (m, 8H), 2.25-2.05 (m, 12H).

Example 55 Preparation ofNI,NI '-((325 '-bis(trzfluoromethyl)-[I,1 '-bl'phenyl]-3,5- diyl)bis(methylene))bis(N3—(3-aminopropyl)pr0pane-I,3-dz'amz'ne), hydrochloride salt (CZ- < ‘NHZ HN—\_/NH H2N4§““1 O OHN CF3 Example 55 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, (3,5-bis(trifluoromethyl)phenyl)boronic acid and utyl (3 -((3- aminopropy1)amino)propyl)carbamate. 1H NMR (500 MHz, D20) 5 8.20 (s, 2H), 8.07 (s, 1H), 7.91 (s, 2H), 7.69 (s, 1H), 4.41 (s, 4H), 3.30 (t, J = 7.5 Hz, 4H), 3.26-3.21(m, 8H), 3.14 (t, J = 8.0 Hz, 4H), 2.23-2.19 (m, 4H), 2.17—2.10 (m, 4H). 13C NMR (125 MHz, D20) 5 140.6, 139.7, 132.4, 131.5, 131.2, 129.9, 127.5, 124.4, 122.2, 50.7, 44.7, 44.6, 44.3, 36.5, 23.7, 22.6. LRMS [M+H]+577.3.

Example 56 Preparation ofNI,NI '-(’(5-(pyridinyl)-I,3-phenylene)bis(methylene))bis(N3-(3- aminopropyl)propane-1,3-diamine), hydrochloride salt (CZ-71) < NH2 HN_\_/NH HN _ HZN HN - 6 HCI Example 56 was ed in a similar fashion to e 45 (CZ-5 8) from 5— bromoisophthaldehyde, pyridin—4-ylboronic acid and tert-butyl (3 -((3- aminopropyl)amin0)pr0pyl)carbamate. 1H NMR (500 MHZ, D20) 5 8.87 (d, J = 6.0 HZ, 2H), 8.39 (d, J = 6.5 Hz, 2H), 8.15 (s, 2H), 7.88 (s, 1H), 4.47 (s, 4H), 3.30 (t, J = 8.0 Hz, 4H), 3.24-3.19(m, 8H), 3.12 (t, J = 8.0 Hz, 4H), 2.24—2.19 (m, 4H), 2.15-2.08 (m, 4H). 13c NMR (125 MHZ, D20)8156.6, 141.6, 1368, 332, 131.0, 125.2, 50.8, 44.9, 44.8, 44.6, 36.7, 23.9, 22.9. LRMS [M+H]+442.4.

Example 57 Preparation ofNI,NI '-(’(5-(6—methoxypyridinyl)-I,3-phenylene)bis (methylene))bis(N3-(3- aminopropyl)propane-1,3-diamine), hydrochloride salt (CZ-72) HN—\_/NH HN —N § \ / 0Me H2N HN -6HC| e 57 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, (6-methoxypyridin-3 -yl)boronic acid and tert—butyl (3-((3- aminopropyl)amino)propyl)carbamate. LRMS [M+H]+ 472.4.

Example 58 Preparation ofNI,N1 '_([1,1 '-biphenyl]-3, 5-dz'ylbz's (methylene))bz’s(N3—(3_((3_ aminopropyl)amino)propyl)propane—I,3-dz'amine), hydrochloride salt ) < HN HN—\_/NH —\—\ [\N -8HC| Example 58 was prepared in a similar n to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, phenylboronic acid and tert—butyl (3-((3-((3- aminopropyl)amino)propyl)amino)propyl)carbamate.

Example 59 Preparation I '—((5-phenoxy-I,3-phenylene)bis(methylene))bis(N3-(3- aminopropyl)propane-1,3-dz'amz'ne), hydrochloride salt (CZ-74) .3” Q

[0478] Step 1: Dimethyl 5-phenoxyts0phthalate. 5-Hydroxyisophthalate (210 mg, 1.00 mmol), phenylboronic acid (0.244 g, 2.0 mmol), copper(Il) acetate (0.182 mg, 1.0 mmol) and triethylamine (0.303 mg, 3.0 mmol) in CH2C12 (20 mL) was stirred at rt in the open air for 38 h. onal phenylboronic acid (0.061 mg, 0.50 mmol) was added, and the reaction mixture was stirred at rt in the open air. After 24 h, additional phenylboronic acid (0.061 g, 0.50 mmol) and copper acetate (0.020 g, 0.11 mmol) were added to the reaction mixture, and the mixture was d for 6 h. The mixture was diluted with CHClz (20 mL) and aqueous HCl (10 mL). The organic layer was separated, washed with aq HCl, aq NaHCOg, dried over MgSO4, and concentrated under reduced re. Purification by column chromatography (10% EtOAc/hexanes) afforded the desired product (0.200 g 70 %) as a white solid. 1H NMR (300 MHz, CDClg) 8 ppm 8.42-8.41 (m, 1H), 7.85 (d, J= 1.5 Hz, 2H), 7.42-7.36 (m, 2H), 7.21-7.16 (m, 1H), 7.05-7.01 (m, 2H), 3.93 (s, 6H).

Step 2: 5—Phen0xyz's0phthalaldehyde. To a solution of dimethyl 5- yisophthalate (0.143 g, 0.50 mmol) in toluene (12 mL) was added drop-wise a solution of Red-Al (60% in toluene, 0.673 mL, 2.0 mmol) and l-methylpiperazine (0.243 mL, 2.2 mmol) in THF at 5 °C, and the resulting mixture was d for 2 h. The on was quenched with H20 (4 mL), and then extracted with EtOAc (20 mL). The organic layer was washed with H20 (10 mL), dried over anhydrous NazSO4, and then concentrated under reduced pressure. Purification by flash column chromatography (10% EtOAc/hexanes) afforded the desired product (0.094 g, 83 %) as a white solid. 1H NMR (300 MHz, CDCl3) 5 ppm 10.04 (s, 2 H), 8.08 (s, 1 H), 7.72 (t, J = 1.5 Hz, 2H), 7.46-7.40 (m, 2H), 7.28-7.24 (m, 1H), 7.09-7.06 (m, 2H).

Step 3: Di—tert—butyl ((((((5-phen0xy—I,3-phenylene)bis(methylene))- bis(azanediyl))bis (propane-3,1-diyl))bis(azanediyl)) bis (propane-3,1-diyl))dicarbamate. To a solution of 5-phenoxyisophthalaldehyde (0.060 g, 0.27 mmol) and 3 A mol. sieves in MeOH, was added N1 inopropyl)-N3-isobutylpropane-1,3-diamine (0.122 g, 0.53 mmol). The resultant mixture was d for 18 h. Sodium borohydride (0.020 g, 0.53 mmol) was added portionwise, and the reaction mixture was stirred for l h. The MeOH was evaporated, and the white solid was taken up in EtOAc (10 mL) and washed with aq. NaOH (10%, 4 mL). The aqueous layer was back—extracted with EtOAc (10 mL). The organic layers were combined, dried over Na2SO4, d and evaporated to afford clear oil.

Methanolic HCl (30 mL, 1.0M) was added to the resultant oil, and the reaction mixture was stirred for 1 h. Evaporation and collection by vacuum filtration afforded a white solid, which was washed hot MeOH (25 mL) to give the desired product (0.064 g, 52%) as a white solid. 1H NMR (500 MHz, D20) 6 7.50 (t, J: 7.5 Hz, 2H), 7.36 (S, 1H), 7.30 (d, J: 7.5 Hz, 1H), 7.24 (s, 2H), 7.17 (d, J: 8.0 Hz, 2H), 4.29 (s, 4H), 3.25—3.18 (m, 12H), 3.14 (t, J: 8.0 Hz, 4H),2.21-2.10 (m, 8H). 13c NMR(125 MHz,D20)6158.2,155.6,133.3,130.3,124.8, 120.5, 119.7, 119.3, 50.5, 44.7, 44.6, 44.2, 36.5, 23.7, 22.6. LRMS [M+H]+457.4.

Example 60 Preparation ofNI,N1'—((4 '-fluoro-[1,1 '-biphenyl]-3,5—diyl)bis(methylene))bis(N3-(3- aminoproprpropane-I,3-diamine), hydrochloride salt (CZ-75) < NH2 |""‘_\_/NH HZNj—liHN O O F '6HCI

[0481] Example 60 was prepared in a similar n to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, (4-fluorophenyl)boronie acid and tert—butyl (3-((3- aminopropyl)amin0)pr0pyl)carbamate. 1H NMR (500 MHZ, D20) 5 7.80 (S, 2H), 7.71-7.68 (m, 2H), 7.56 (s, 1H), 7.25 (t, J = 9 Hz, 2H), 4.37 (s, 4H), 3.26 (t, J = 8 Hz, 4H), 3.19 (q, J = 8 Hz, 8H), 3.11 (t, J = 7.5 Hz, 4H), 2.22—2.07 (m, 8H). 13c NMR (125 MHz, D20) 8 162.9 (d, 244 Hz), 135.3(d, 3 Hz),132.3,130.1,129.7,129.1(d,8 Hz), 116.1 (d, 21 Hz), 51.1, 44.9, 44.8, 44.4, 36.7, 23.9, 22.9. LRMS [M+H]+ 459.4.

Example 61 ation ofNI,N1’—([I,I'-biphenyl]-3,5-diylbis(methylene))bis(propane-I,3-diamine), hloride salt (CZ-77) HN—\_/NH2 “2“) O O -4HCI Example 61 was prepared in a similar fashion to Example 45 (CZ-5 8) from 5- bromoisophthaldehyde, phenylboronic acid and tert—butyl (3-amin0propyl)carbamate. N1,N1'- -biphenyl]—3,5—diylbis(methylene))bis(propane-1,3-diamine). 1H NMR (300 MHZ, D20) 8 7.82 (s, 2H), 7.71-7.64 (m, 2H), 7.54-7.41 (m, 4H), 4.34 (s, 4H), 3.22 (t, J: 7.8 Hz, 4H), 3.08 (t, J: 7.5 Hz, 4H), 2.11 (quint, J: 7.5 Hz, 4H).

Example 62 Preparation ofNI,N1 '—((5-butoxy-I,3-phenylene)bis(methylene))bis(N3-(3- aminopropybpropane-1,3-a’z'amz'ne), hydrochloride salt (CZ-76) < NH2 HN—\_/NH HN_? /_/_ - 6 HCI HZN HN

[0483] Step 1: Dimethyl 5-butoxyz'sophthalate. Dimethyl oxyisophthalate (0.40 g, 1.90 mmol), cesium carbonate (1.30 g, 3.80 mmol) and CH3CN (12 mL) were added to a round-bottom flask and stirred for 15-30 min. tane (0.42 g, 2.28 mmol) was added,and the on was stirred for 16 h. The solvent was concentrated under reduced pressure, and the reaction mixture was partitioned between EtOAc (50 mL) and H20 (50 mL). The organic layer was separated and the aqueous layer extracted with EtOAc (50 mL).

The combined organics were dried over NaZSO4, filtered, and concentrated under reduced pressure to afford the desired product which was used t further purification.

Step 2: (5-Butoxy-I,3-phenylene)a’imethanol. To a solution of dimethyl 5- butoxyisophthalate (0.51 g, 1.90 mmol) in THF (16 mL) was slowly added LiAlH4 (0.40 g, 10.5 mmol). The reaction e was stirred for 8 h. The reaction was quenched by the addition of 2N aq. HCl (5 mL). The mixture was extracted with EtZO (2 x 25 mL) and EtOAc (2 x 25 mL), the organics layers combined, dried over Na2S04, filtered, and concentrated under reduced pressure to afford the d product (0.24 g) as an oil, which was used without further ation.

[0485] Step 3: 5-Butoxyisophthalala’ehya’e. (5-Butoxy-l,3-pheny1ene)dimethanol (0.24 g, 1.15 mmol) and CH2C12 (10 mL) were added to a round-bottom flask. To the solution was added PCC (0.74 g, 3.45 mmol), and the reaction was stirred at rt for 16 h. The reaction mixture was concentrated under reduced pressure. Purification by column chromatography (90% s/EtOAc) afforded the desired product (0.18 g, 44%, 3 steps) as a white semi- solid. 1H NMR (300 MHz, CDC13) 5 ppm 9.99 (s, 2H), 7.89 (s, 1H), 7.58 (s, 2H), 4.03 (t, J: 6.6 Hz, 2H), 1.77 (quint, J= 6.6 Hz, 2H), 1.47 (sext, J= 7.2 Hz, 2H), 0.94 (t, J= 7.5 Hz, 3H). 13C NMR (75 MHz, CDC13)5 ppm191.3,160.6,138.6,124.2,120.1, 68.9, 313,194,141.

Step 4: Di-tert—butyl ((((((5-butoxy-1,3-phenylene)bis(methylene))bis(azanediyl))- bis(propane-3, I—diyl))bis(azanediyl)) bis(propane—3, I—diyl))dicarbamate. 5- Butoxyisophthalaldehyde (0.80 g, 0.85 mmol) and MeOH (15 mL) were added to a round- bottom flask. To the solution was added tert-butyl (3-((3-aminopropyl)amino)propyl)— carbamate (0.39 g, 1.70 mmol), and the reaction mixture was stirred at rt for 24 h. Sodium borohydride (0.13 g, 3.40 mmol) was added and the reaction e stirred for l h. The reaction mixture was concentrated under reduced pressure and aq. NaOH (10%, 50 mL) and EtOAc (50 mL) were added. The layers were separated and the aqueous layer was extracted with EtOAc (50 mL). The organic layers combined, dried over Na2804, filtered, and concentrated under reduced pressure to afford the desired product as a clear oil which was used without further purification.

Step 5: -((5—Butoxy—I,3-phenylene)bis(methylene))bis(N3-(3- aminopropyl)propane-1,3-diamine), hydrochloride salt. To the crude di-tert—butyl ((((((5- butoxy- l ,3 -phenylene)bis(methylene))bis(azanediyl))bis(propane-3 , l -diyl))bis(azanediyl))- opane-3,1-diyl))dicarbamate from Step 4 was added methanolic HCl (50 mL, 1.0 M).

The reaction mixture was stirred for 2 h and concentrated under reduced pressure. The solid was collected by vacuum filtration and washed with Et20 (10 mL) and hot MeOH (10 mL) to afford the d product (0.27 g, 48%) as a white solid. 1H NMR (500 MHz, D20) 8 ppm 7.20 (s, 3H), 4.30 (s, 4H), 4.15 (t, J= 6.5 Hz, 2H), 3.26-3.20 (m, 12H), 3.14 (t, J= 7.5 Hz, 4H), 2.22-2.10 (m, 8H), 1.79 (quint, J: 6.5 Hz, 2H), 1.48 (sext, J: 7.5 Hz, 2H), 0.96 (t, J = 7 Hz, 3H). 13C NMR (125 MHz, D20) 159.2, 132.9, 123.5,117.3, 68.8, 50.8, 44.7, 44.6, 44.1, 36.5, 30.3, 23.7, 22.6, 18.5, 13.0. LRMS [M+H]+437.4.

Example 63 ation ofN],NI '-((5-((2-ethylhexyl)oxy)-1, 3-phenylene)bis(methylene))bis(N3-(3- aminopropyl)propane—1,3—diamine), hloride salt (CZ—8 l) < NH2 HN_\_/NH j?HN 0 Me H2N HN - 6 HCI Me Example 63 was prepared in a similar fashion to Example 62 (CZ-76) from dimethyl xyisophthalate, 3-(bromomethyl)heptane and tert-butyl (3 -((3- aminopropyl)amino)propyl)carbamate. LRMS [M+H]+ 493.5.

Example 64 Preparation ofN1,Nl'-(’(5-(2-ethylbutoxy)-1, 3-phenylene)bis(Methylene))bis(N3-(3- aminoproprpropane-I,3-diamine), hydrochloride salt (CZ-90) < NH2 HN_\_/NH RHN we HZN HN Me - 6 HCI e 64 was prepared in a similar fashion to Example 62 (CZ-76) from dimethyl 5-butoxyisophthalate, 3-(bromomethyl)pentane and tert-butyl (3 -((3- aminopropyl)amino)propyl)carbamate. 1H NMR (500 MHZ, D20) 8 7.23 (s, 2H), 7.22 (s, 1H), 4.32 (s, 4H), 4.06 (t, J: 7.0 HZ, 2H), 3.27-3.18 (m, 12H), 3.15 (t, J: 8.0 HZ, 4H), 2.24- 2.12 (m, 8H), 1.72 (m .1: 6.0 HZ, 1H), 1.52—1.44 (m, 4H), 0.94—0.91 (m, 6H). 13C NMR (125 MHZ, D20) 8 159.6, 132.9, 123.5, 117.5, 71.43, 50.8, 44.7, 44.6, 44.1, 40.1, 36.5, 23.7, 22.6, 10.3. LRMS [M+H]+465.4.

Example 65 Preparation ofNI,N1'-(’(5-(Benzyloxy)-1,3-phenylene)bis(methylene))bis(N3-(3— aminopropyl)propane-1,3-a’iamine), hydrochloride salt (CZ-95) < NH2 l""l_\_/NH §lHN °L© H2” HN - 6 HCI Example 65 was prepared in a similar fashion to e 62 (CZ-76) from dimethyl xyisophthalate, benzyl e and tert—butyl (3-((3- aminopropyl)amino)propyl)carbamate. 1H NMR (500 MHZ, D20) 8 7.52 (d, J = 7.0 HZ, 2H), .41 (m, 3H), 7.23 (s, 2H), 7.20 (s, 1H), 5.23 (s, 2H), 4.29 (s, 4H), 3.22—3.15 (m, 12H), 3.13 (t, J = 8.0 Hz, 4H), 2.20—2.09 (m, 8H). 13c NMR (125 MHz, D20) 8 158.7, 136.0, 133.0, 132.5, 128.8, 128.5, 128.0, 123.9, 117.7, 70.4, 50.7, 44.7, 44.6, 44.3, 44.0, 36.5, 23.7, 22.6.

LRMS [M+H]+ 471.4.

Example 66 Preparation ofNI,N1 '—('(5-(cyclohexylmethoxy)-1,3-phenylene)bis(methylene))bis (N3-(3- aminopropyl)propane-1,3-diamine), hydrochloride salt (CZ-101) < NH2 HN_\_/NH HZN HN ' 6 HCI Example 66 was ed in a similar n to Example 62 (CZ-76) from dimethyl 5-butoxyisophthalate, (bromomethyl)cyclohexane and tert—butyl (3-((3- aminopropy1)amin0)propyl)carbamate. 1H NMR (500 MHZ, D20) 8 7.16 (d, J = 7.5 Hz, 3H), 4.27 (s, 4H), 3.93 (t, J = 6.0 Hz, 2H), 3.23-3.09 (m, 16H), 2.19-2.07 (m, 8H), 1.83-1.66 (m, 6H), 1.31-1.17 (m, 3H), 1.10-1.05 (m, 2H). 13C NMR (125 O)8131.4, 131.2, 131.1,130.2, 50.8, 47.9, 44.6, 446,442, 44.1, 30.9, 28.1, 28.1, 25.6, 25.4, 22.6, 21.9, 13.3.

Example 67 Preparation ofNI,N1 '-([1, I ’-biphenyl]-3, 5-diylbis (methylene))bis(N3-(3- (butylamino)propyl)propane-1,3-a’iamine), hydrochloride salt (CZ-83) 6...my”)...0 .0 Step 1: [1,1'-Biphenyl]-3,5-dicarbala’ehya’e. A solution of 5- bromoisophthaldehyde (40.0 g, 187.8 mmol), phenylboronic acid (22.9 g, 187.8) and potassium carbonate (64.8 g, 469.5 mmol) in DME/HZO (5:1 600 mL) was purged with N2 for 5 min. Tetrakispalladium triphenylphosphine (1.1 g mg, 0.9 mmol) was added and the reaction e was heated to 90 CC and stirred for 16 h. The reaction mixture was cooled, filtered through a pad of celite and the solvent evaporated under reduced pressure.

Purification by column chromatography (10% EtOAc/hexanes) afforded the desired product (74%, 29.1 g) as a tan solid. 1H NMR (300 MHz, CDClg) 6 ppm 10.18 (s, 2H), 8.37 (d, J= 1.2 Hz, 2H), 8.35 (t, J: 1.5 Hz, 1H), 7.70-7.66 (m, 2H), .43 (m, 3H).

Step 2: N1,N1'-([1,1'-biphenyl]-3,5-diylbis(methylene))bis(N3-(3- (bugzlamino)pr0pyl)pr0pane-I,3-a’z‘amz’ne). [1,1'-Biphenyl]—3,5-dicarbaldehyde (0.66 g, 3.15 mmol) and MeOH (50 mL) were added to a round-bottom flask. To the solution was added N1-(3-aminopropyl)-Z\73-butylpropane-l,3-diamine (1.18 g, 6.31 mmol) and the reaction e was stirred at rt for 24 h. Sodium borohydride (0.48 g, 12.62 mmol) was added and the reaction mixture stirred for 1 h. The reaction mixture was concentrated under d pressure to afford a white solid. Aq. NaOH (10%, 100 mL) and EtOAc (100 mL) were added and the reaction mixture stirred for 1 h. The layers were separated and the aqueous layer was extracted with EtOAc (100 mL). The organic layers were combined, dried over NaZSO4, filtered, and concentrated under reduced pressure to afford a clear oil which was used without further purification.

Step 3: N’,N’ '_([1,1 '-bz'phenyl]-3, 5-diylbis(methylene))bz’s(N3-(3- (butylamin0)pr0pyl)pr0pane-I,3-dz'amz'ne) hydrochloride salt. To the crude N1,N1'—([1,1‘— yl]—3,5-diylbis(methylene))bis(Z\73-(3-(butylamino)propyl)propane-1,3-diamine) from step 2 was subjected to acidification with methanolic HCl (100 mL, 1.0M). The reaction mixture stirred at rt for 2 h. The reaction e was concentrated under reduced pressure and the solid was collected by vacuum filtration and washed with Et20 (50 mL) and hot MeOH (50 mL) to afford the desired product (1.24 g, 51%) as a white solid. 1H NMR (500 MHz, D20) 5 7.83 (s, 2H), 7.71 (d, J = 7.5 Hz, 2H), 7.55—7.52(m, 3H), 7.49—7.45 (m, 1H), 4.35 (s, 4H), 3.26 (t, J = 7.5 Hz, 4H), .16 (m, 8H), 3.14 (t, J = 8.0 Hz, 4H), 3.05 (t, J = 8.0 Hz, 4H), .10 (m, 8H), 1.65 (quint, J = 7.0 Hz, 4H), 1.37 (sext, J = 8.0 Hz, 4H), 0.91 (t, J = 7.0 Hz, 6H). 13C NMR (125 MHz, D20) 5 142.5, 138.9, 132.3, 130.3, 129.8, 129.5, 128.7, 127.3, 51.0, 47.8, 44.9, 44.5, 44.4, 27.7, 22.9, 22.9, 19.3, 13.0. LRMS [MJrH]+ 553.5.

Example 68 Preparation ofN’Jv’ '—([1,1 '-bz'phenyl]-3, 5-dz'ylbz's (methylene))bz's(N3-(3— (isobutylamindpropybpropane-I,3-dz'amine), hydrochloride salt ) HNLQNy 3"? O O l-gli HN ' 6 HCI Method 1 Step 1: [1,1 '—Bz‘phenyl]—3,5—dicarbaldehyde. A solution of 5— bromoisophthaldehyde (40.0 g, 187.8 mmol), phenylboronic acid (22.9 g, 187.8) and potassium carbonate (64.8 g, 469.5 mmol) in DME/HZO (5:1 600 mL) was purged with N2 for 5 min. Tetrakispalladium triphenylphosphine (1.1 g mg, 0.9 mmol) was added and the reaction mixture was heated to 90 CC and d for 16 h. The reaction mixture was , filtered through a pad of celite and the solvent evaporated under reduced pressure.

Purification by column chromatography (10% EtOAc/hexanes) afforded the desired product (74%, 29.1 g) as a tan solid. 1H NMR (300 MHZ, CDClg) 8 ppm 10.18 (s, 2H), 8.37 (d, J= 1.2 Hz, 2H), 8.35 (t, J: 1.5 Hz, 1H), .66 (m, 2H), 7.55-7.43 (m, 3H).

[0496] Step 2: t—butyl ((((([1,1 henyl]-3,5-dl'ylbl's(methylene))bis(azanediy0)- bis(pr0pane-3, l-dbzwbismzanediyl»bis(pr0pane—3, 1-diyl))dicarbamate. [1 1 '-Biphenyl]- 3,5-dicarbaldehyde (3.62 g, 17.2 mmol, 1 ) and MeOH (100 mL) were added to a round- bottom flask. To the solution was added tert—butyl (3-((3-aminopropyl)amino)propyl)- carbamate (7.96 g, 34.4 mmol), and the reaction mixture was stirred at rt for 24 h. Sodium borohydride (2.62 g, 69.0 mmol) was added and the reaction e stirred for 1 h. The reaction mixture was concentrated under reduced pressure to afford a white solid. Aqueous NaOH (10%, 200 mL) and EtOAc (200 mL) were added, and the reaction mixture was stirred for 1 h. The layers were separated, and the aqueous layer was extracted with EtOAc (200 mL). The organic layers were combined, dried over , filtered, and concentrated under reduced pressure to afford a clear oil that was used without further purification.

Step 3: N1,N1'-([1,1'-bz'phenyl]-3,5-dl'ylbz's(methylene))bis(N3-(3- aminopropyl)pr0pane-1,3-diamine), hydrochloride salt. To the crude di-tert—butyl ((((([1,1'- biphenyl]-3 ,5-diylbis(methylene))bis(azanediyl))bis(propane-3 ,1-diyl))bis(azanediyl))- bis(propane-3,1-diyl))dicarbamate from Step 2 was subjected to acidification with methanolic HCl (200 mL, 1.0M). The reaction mixture stirred at rt for 2 h. The reaction mixture was concentrated under reduced pressure, and the solid was collected by vacuum filtration and washed with Et20 (50 mL) and hot MeOH (50 mL) to afford the desired product (6.8 g, 60%) as a white solid. 1H NMR (500 MHZ, D20) 8 7.85 (s, 2H), 7.74 (d, J : 7.5 Hz, 2H), 7.59-7.56 (m, 3H), .49 (m, 1H), 4.38 (s, 4H), 3.29 (t, J = 8.0 Hz, 4H), 3.23 (q, J = 5.0 Hz, 8H), 3.14 (t, J = 8.0 Hz, 4H), 2.25-2.19 (m, 4H), 2.17-2.11 (m, 4H). LRMS [M+H]+ 441.4.

Step 4: N1,N1'-([1,1'-Bz‘phenyl]-3,5-dz'ylbz's(methylene))bis(N3-(3- (isobutylamz'no)pr0pyl)pr0pane-I,3-diamine). To N1,N1'-([1,1‘-biphenyl]-3,5- diylbis(methylene))bis(1\73-(3-aminopropyl)propane-1,3-diamine), hydrochloride salt was added aq. NaOH (10%, 100 mL) and 75% CHClg/i-propanol (100 mL). The layers were separate and the aqueous layer was extracted with 75% CHClg/i-propanol (4 x 100 mL). The organic layers were ed dried over Na2804, filtered, and concentrated under reduced pressure to afford a clear oil which was used without r ation.

To a round-bottom flask was added the crude N1,N1'-([1,1'-biphenyl]-3,5- diylbis(methylene))bis(N3-(3-aminopropyl)propane—1,3—diamine) (3.5 g, 8.0 mmol) and MeOH (50 mL). To the solution was added isobutyraldehyde (1.15 g, 15.9 mmol) and the reaction mixture was stirred at rt for 24 h. Sodium borohydride (1.2 g, 31.9 mmol) was added and the reaction mixture was stirred for 1 h. The reaction mixture was concentrated under reduced pressure to afford a white solid. Aq. NaOH (10%, 200 mL) and EtOAc (200 mL) were added, and the mixture was stirred for 1 h. The layers were separated and the aqueous layer was extracted with EtOAc (200 mL). The combined organics were dried over Na2804, filtered, and concentrated under reduced pressure to afford the desired t as a clear oil which was used without further ation.

Step 5: N1,N1'-([1,1'-Bl'phenyl]-3,5-dl'ylbl's(methylene))biS(N3-(3- (isobutylamz'no)pr0pyl)pr0pane—I,3—diamine), hloride salt. To the crude —([1,1'- biphenyl]—3,5-diylbis(methylene))bis(N3-(3-(isobutylamino)propyl)propane-1,3-diamine) was added methanolic HCl (200 mL, 1.0M). The reaction mixture was stirred at rt for 1 h. The reaction e was concentrated under reduced pressure, and the solid was collected by vacuum filtration and washed with EtzO (50 mL) and hot MeOH (50 mL) to afford the desired product as a white solid (3.07 g, 50%). 1H NMR (500 MHz, D20) 8 7.87 (s, 2H), 7.75 (d, J= 7.5, 2H), 7.59-7.48 (m, 4H), 4.39 (s, 4H), 3.26 (t, J= 8.0 Hz, 4H), 3.21-3.12 (m, 12H), 2.92 (d, J: 7.0 Hz, 4H), 2.21-2.10 (m, 8H), 2.01 (sept, J: 7.0 Hz, 2H), 0.99 (d, J: 7.0 Hz, 12H). 13C NMR(125 MHz, D20)8142.8,139.1,132.4,130.1,129.8,129.5,128.7, 127.3, 55.1, 51.1, 44.9, 44.8, 44.4, 25.8, 22.8, 22.7, 19.2. LRMS [M+H]+ 553.5.

Method 11 Step 1: [1,1’-Bz'phenyl]-3,5-dz'carbaldehyde. A solution of 5- bromoisophthaldehyde (40.0 g, 187.8 mmol), phenylboronic acid (22.9 g, 187.8) and ium carbonate (64.8 g, 469.5 mmol) in DME/HzO (5:1, 600 mL) was purged with N2 for 5 min. Tetrakispalladium triphenylphosphine (1.1 g mg, 0.9 mmol) was added and the reaction mixture was heated to 90 OC and stirred for 16 h. The reaction mixture was cooled, filtered through a pad of Celite diatomaceous earth, and the solvent was evaporated under reduced pressure. ation by column chromatography (10% hexanes) afforded the desired t (74%, 29.1 g) as a tan solid. 1H NMR (300 MHz, CDC13) 8 ppm 10.18 (s, 2H), 8.37 (d, J= 1.2 Hz, 2H), 8.35 (t, J= 1.5 Hz, 1H), 7.70-7.66 (m, 2H), 7.55-7.43 (m, 3H).

Step 2: N1,N1'-([1,1'-Bz'phenyl]-3,5-diylbis(methylene))bis(N3-(3- (isobutylamino)pr0pyl)pr0pane-I,3-dz'amine). [1,1'-Biphenyl]—3,5-dicarbaldehyde (3.03 g, 14.4 mmol) and MeOH (60 mL) were added to a round—bottom flask. To the solution was added N1-(3-aminopropyl)-]\]3-isobutylpropane-1,3-diamine (5.40 g, 28.8 mmol), and the reaction mixture was stirred at rt for 24 h. Sodium dride (0.55 g, 14.4 mmol) was added, and the reaction mixture was stirred for 2 h. The reaction mixture was concentrated under reduced pressure to afford a white solid. Aqueous NaOH (10%, 50 mL) was added, and the reaction mixture stirred for 30 min. The solution was extracted with CHC13 (3 x 75 mL). The c layers were combined, dried over Na2S04, filtered, and trated under reduced pressure to afford a clear oil which was used without further purification.

Step 3; N1,N1'-([1,1'-Biphenyl]—3,5-diylbis(methylene))bis(]\]3-(3- (isobutylamino)propyl)propane-1,3-diamine) hydrochloride salt: To the crude N1,N1'-([l,l'- biphenyl]—3,5—diylbis(methylene))bis(1\73-(3-(isobutylamino)propyl)propane—1,3—diamine) from step 2 was added methanolic HCl (75 mL, 2.0M). The resulting white solid was collected by vacuum ion and washed with minimal MeOH and EtzO and stored under high vacuum to afford the desired product (9.3 g, 89%) as a white solid.

Example 69 Preparation of3, 5-bis(((3-((3-aminopropyl)amino)propyl)amino)methyl)phenol, hydrochloride salt (CZ-87) < NHZ HN_'\_/NH j?HN 0H . 6 HCI HZN HN

[0504] Example 69 was prepared in a similar fashion to Example 62 (CZ-76) from dimethyl -butoxyis0phthalate and tert—butyl (3 -((3-aminopropyl)amino)pr0pyl)carbamate.

Example 70 Preparation of(3 C5 '-Bis(((3- ((3-aminopropyl)amino)propyl)amino)methyl)—[I, I enyl]- 4-yl)(piperidin-I—ybmethanone, hydrochloride salt (CZ-88) < NH2 HzNj—lil"N O O 2 -6HC| O Example 70 was ed in a similar fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, din- l -yl(4-(4,4,5 ,5 —tetramethyl— l ,3 ,2—di0xaborolan—2—yl)- phenyl)methanone and tert—butyl (3-((3-aminopr0pyl)amino)propyl)carbamate. 1H NMR (300 MHz, D20) 5 7.87 (s, 2H), 7.78 (d, J = 8.4 Hz, 2H), 7.58 (s, 1H), 7.54—7.49 (m, 2H), 4.37 (s, 4H), 3.68 (bs, 2H), 3.42—3.38 (m, 2H), 3.26—3.13 (m, 12H), 3.07 (t, J = 7.8 Hz, 4H), 2.20—2.01 (m, 8H), 1.71-1.64 (m, 4H), 1.58-1.49 (m, 4H). LRMS [M+H]+552.4.

Example 71 Preparation ofI, I '-((([1,1 '-biphenyl]-3,5diylbis(methylene))bis(azanediyljjbismropane-iI- diyl))bis(tetrahydropyrimidin-2(II-D-one), hydrochloride salt (CZ-89) ° 2 HCI HM "WHOO—\_/:HN—<

[0506] Example 71 was prepared in a similar fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, phenyl boronic acid, 1-(3-aminopropyl)tetrahydropyrimidin—2(1H)- one and tert—butyl (3-((3-aminopropyl)amin0)pr0pyl)carbamate. 1H NMR (300 MHz, D20) 8 7.80 (d, J = 6.5 HZ, 2H), 7.71-7.68 (m, 2H), .45 (m, 4H), 4.30 (s, 4H), 3.34 (t, J = 6.6 Hz, 4H), 3.21 (t, J = 6.0 Hz, 4H), 3.11 (t, J = 6.0 Hz, 4H), 3.04 (t, J = 7.2 HZ, 4H), .88 (m, 4H), 1.78 (p, J = 6.0 Hz, 4H). LRMS 493.4.

Example 72 Preparation ofNI,NI '-([I, I enyl]-3,5-diylbis(methylene))bis(N3-(3-(octylamino)propyl)- propane-1,3-diamine), hydrochloride salt (CZ-92) HN—\_/NH HNA§ H <:/—\

[0507] Example 72 was prepared in a similar fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, phenylboronic acid, tert-butyl (3 -((3-amin0propyl)amino)propyl)— carbamate, and octanal. 1H NMR (500 MHZ, D20) 8 7.88 (s, 2H), 7.75 (d, J = 8.0 Hz, 2H), 7.60-7.56(m, 3H), .50 (m, 1H), 4.40 (s, 4H), 3.26 (t, J = 8.0 Hz, 4H), 3.21-3.16 (m, 8H), 3.14 (t, J = 8.0 Hz, 4H), 3.06 (t, J = 8.0 Hz, 4H), 2.21-2.09 (m, 4H), 1.69-1.65 (m, 4H), 1.38—1.28 (m, 20H), 0.86 (t, J = 6.0 Hz, 4H). 13C NMR (125 MHz, D20) 8 142.5, 138.8, 132.2, 129.9, 129.5, 129.3, 128.5, 127.0, 50.8, 47.8, 44.6, 44.1, 30.9, 28.1, 28.0, 25.6, 25.4, 22.7, 22.6, 21.9, 13.3. LRMS [M+H]/2+333.3.

Example 73 Preparation ofN’,N’ '—([1,1 '-bz'phenyl]-3, 5-dz'ylbz's (methylene))bz's(N3-(3— (benzylamino)propyl)propane-1,3-diamine), hloride salt (CZ-94) HN5mO H?NH - 6 HCI

[0508] Example 73 was prepared in a similar fashion to e 68 ) from 5- bromoisophthaldehyde, phenylboronic acid, tert-butyl (3-((3-aminopropyl)amin0)pr0pyl)- carbamate, and benzaldehyde. 1H NMR (300 MHZ, D20) 5 7.84 (s, 2H), 7.74-7.70 (m, 2H), 7.58-7.50 (m, 4H), 7.47 (s, 10H), 4.37 (s, 4H), 4.24 (s, 4H), 3.26-3.11 (m, 16H), 2.20—2.05 (m, 8H).

Example 74 Preparation ofNI,N1 '-([1, I '-bz'phenyl]-3,5-dl'ylbz's(methylene))bis(lV3-(3- ((Cyclohexylmethyl)amino)propyl)propane-I,3-dz'amz'ne), hydrochloride salt (CZ-96) (Tin Example 74 was prepared in a similar fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, phenylboronic acid and N1—(3—amin0pr0pyl)-NS— (cyclohexylmethyl)pr0pane-l,3-diamine. 1H NMR (500 MHZ, D20) 5 7.87 (s, 2H), 7.75 (d, J = 7.5 Hz, 2H), 7.59-7.56(m, 3H), 7.51—7.49 (m, 1H), 4.40 (s, 4H), 3.28 (t, J = 8.0 Hz, 4H), 3.24-3.18 (m, 8H), 3.15 (t, J = 8.0 Hz, 4H), 2.93 (d, J = 7.0 Hz, 4H), 2.24—2.12 (m, 8H), 1.74- 1.60 (m, 12H), .14 (m, 6H), 1.05-0.98 (m, 4H). 13C NMR (125 MHz, D20) 5 142.4, 138.8, 132.1, 130.0, 129.6, 129.3, 128.5, 127.0, 53.7, 50.8, 44.7, 44.6, 44.2, 34.5, 29.7, 25.4, 24.9, 22.6, 22.5. LRMS [M+H]+633.5.

Example 75 Preparation ofNI,N1 '-((5-(I-methyl—IH—pyrazol—4-yl)-I,3-phenylene)bis(methylene))bis(N3- (3-(isobutylamino)propyl)propane-I, 3—diamine), hydrochloride salt (CZ-102) (—H‘N HN—\_/NH —>— H?HN {Ill HN HN - 6 HCI

[0510] Example 75 was prepared in a r fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, (1-methyl-1H-pyrazolyl)boronic acid and tert-butyl (3-((3- aminopropyl)amino)propyl)carbamate. 1H NMR (300 MHZ, D20) 5 7.75—7.69 (m, 4H), 6.60 (d, J = 2.1 Hz, 2H), 4.38 (s, 4H), 3.92 (s, 3H), 3.27-3.09 (m, 16H), 2.89 (d, J = 7.2 Hz, 4H), 2.21-2.06 (m, 8H), 2.03-1.94 (m, 2H), 0.97 (d, J = 6.9 Hz, 12H). 1H NMR (300 MHz, D20) 5 7.75-7.69 (m, 4H), 6.60 (d, J = 2.1 Hz, 2H), 4.38 (s, 4H), 3.92 (s, 3H), 3.27-3.09 (m, 16H), 2.89 (d, J = 7.2 Hz, 4H), 2.21-2.06 (m, 8H), 2.03-1.94 (m, 2H), 0.97 (d, J = 6.9 Hz, 12H). e 76 Preparation ofNI,N1/—([I,I '-biphenyl]-3,5-diylbis(methylene))bis(N4-(3- (isobutylamino)propyl)butane-1,4-diamine), hydrochloride salt (CZ-1 11) .5 .. H9 § O O_\_/_ Example 76 was prepared in a similar fashion to Example 76 (CZ-86) from 5- bromoisophthaldehyde, phenylboronic acid and N1-(3-amin0pr0pyl)-]\73-is0butylpropane-1,3- diamine. 1H NMR (500 MHz, D20) 6 7.86 (s, 2H), 7.75 (d, J = 7.5 Hz, 2H), 7.58 (t, J = 8.0 Hz, 3H), 7.50 (t, J = 7.5 Hz, 1H), 4.37 (s, 4H), 3.20-3.11 (m, 16H), 2.92 (d, J = 7.5 Hz, 4H), .10 (m, 4H), 2.06-1.97 (m, 2H), 1.88-1.80 (m, 8H), 0.99 (d, J = 6.5 Hz, 12H). 13C NMR(125 MHz, D20) 6 142.7, 139.1, 132.6, 130.1, 129.7, 129.5, 128.7, 127.7, 55.1, 50.9, 47.2, 46.7, 44.5, 44.7, 25.8, 23.0, 22.9, 22.7, 19.2. LRMS [M+H]+ 581.5.

Example 77 Preparation 0fN1,NIV—([I,I '-bz'phenyl]-3,5-diylbz's(methylene))bz'S(N3-(3- (z'sabutylamz'no)pr0pyl)-2,2-a’imethylpr0pane-I,3-a’z'amz'ne), hloride salt (CZ-112) WFQNWF x0 O >_/ -6HC| Example 77 was prepared in a similar n to Example 68 (CZ-86) from 5- bromoisophthaldehyde, phenylboronic acid and N1-(3-(is0butylamino)propyl)—2,2- dimethylpropane-l,3-diamine. NMR (500 MHz, D20) 8 7.89 (s, 2H), 7.71 (d, J = 7.5 HZ, 2H), 7.62 (s, 1H), 7.52 (t, J = 7.5 Hz, 2H), 7.44 (t, J = 7.5 Hz, 1H), 4.39 (s, 4H), 3.19-3.01 (m, 16H), 2.89 (d, J : 7.5 Hz, 4H), 2.21-2.15 (m, 4H), 2.03-1.96 (m, 2H), 1.18 (s, 12H), 0.97 (d, J = 6.5 Hz, 12H). 13c NMR (125 MHz, D20) 5 1422, 1388,1317, 1314, 130.2, 129.5, 128.7, 127.2, 55.6, 55.1, 4.9, 52.1, 46.3, 45.0, 33.2, 25.8, 22.5, 22.4, 219,193. LRMS [M+H]+609.5.

Example 78 Preparation ofNI,N1 '-([I, I '-bipheriyl]-3,5-diylbis(methylene))bis(lV3-(3— (hexylamino)propyl)proparie-I,3-diamine), hydrochloride salt (CZ-l 14)

[0513] Example 78 was prepared in a similar fashion to Example 68 (CZ-86) from 5- sophthaldehyde, phenylboronic acid and amin0propyl)-N3-hexylpropane-1,3- diamine. 1H NMR (500 MHz, D20) 8 7.83 (s, 2H), 7.71 (d, J = 7.5 Hz, 2H), .53 (m, 3H), 7.47 (t, J = 6.5 Hz, 1H), 4.34 (s, 4H), 3.25 (t, J =8 Hz, 4H), 3.19 (q, J = 8 Hz, 8H), 3.13 (t, J = 8 Hz, 4H), 3.04 (t, J = 8.5 Hz, 4H), 2.22-2.09 (m, 8H), 1.66 (p, J = 7.5 Hz, 4H), 1.36- 1.31 (m, 4H), 1.28-1.27 (m, 8H), 0.85 (t, J = 7 Hz, 6H). 13C NMR (125 MHz, D20) 8 142.5, 139.0, 132.3, 130.3, 129.8, 129.5, 128.7, 127.3, 51.0, 48.1, 44.9, 44.4, 44.4, 30.6, 25.6, 25.5, 22.9, 22.8, 21.9, 13.5. LRMS [M+H]+ 609.5.

Example 79 Preparation ofNI,N1 '— ([1, I '-bipheriyl]-3, 5-diylbis(methylerie))bis(dodecane-1, I2-diamine), hydrochloride salt (CZ-115) Example 79 was prepared in a similar fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, phenylboronic acid and tert—butyl (12-amin0dodecyl)carbamate. 1H NMR (500 MHZ, D20) 8 7.92 (s, 2H), 7.80 (d, J = 8.5 Hz, 2H), 7.61—7.64 (m, 3H), 7.57—7.54 (m, 1H), 4.41 (s, 4H), 3.10 (t, J =75 Hz, 4H), 3.03 (t, J = 7.5 Hz, 4H), 1.77-1.66 (m, 8H), 1.39-1.36 (m, 12H), 1.35-1.28 (m, 22H). 13c NMR (125 MHz, D20) 8 142.5, 139.0, 132.3, 130.3, 129.8, 129.5, 128.7, 127.3, 51.0, 48.1, 44.9, 44.4, 44.4, 30.6, 25.6, 25.5, 22.9, 22.8, 21.9, 13.5. LRMS [M+H]+ 579.5.

Example 80 Preparation I '—([I, I '-bz'phenyl]-3, 5-dl'ylbz's (methylene))bis(NIZ—isobugildodecane-I, 12- diamz'ne), hydrochloride salt (CZ-116) iO 11 "1 .4HCI Example 80 was prepared in a similar fashion to Example 68 (CZ-86) from 5— bromoisophthaldehyde, boronic acid and N1-isobutyldodecane-1 12-diamine. 1H NMR, (500 MHz, D20 + CDgOD) 8 7.90 (s, 2H), 7.76 (d, J = 7.5 Hz, 2H), 7.60-7.56 (m, 3H), 7.52 (t, J = 7.5 Hz, 1H), 4.38 (s, 4H), 3.04 (t, J :80 Hz, 4H), 3.01 (t, J = 8.5 Hz, 4H), 2.88 (d, J = 7.5, 4H), 2.06-1.98 (m, 2H), 1.74-1.65 (m, 8H), 1.33-1.30 (m, 12H), 1.28-1.23 (m, 20H), 1.01 (d, J = 7.5, 12H). 13C NMR (125 MHz, D20 + CD30D)8142.5, 138.8, 132.4, 130.1, 129.5, 129.3, 128.5, 127.0, 51.1, 46.6, 39.5, 28.6, 28.5, 28.4, 28.2, 28.0, 26.7, 25.6, 25.6, 25.2.

LRMS [M+H]+ 691.6.

Example 81 Preparation ofNI,N1',N1H-([I, I '-biphenyl]-3,3 ',5—trz'yltris (methylene))tris(N12- z'sobulyldoa’ecane-I, I2-a’iamz'ne) (CZ-1 17) NH HN H H ”A NH ”A Y” -6HC|

[0516] e 81 was prepared in a r fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, 3-f0rmyl-phenylb0r0nic acid and N1-is0butyld0decane-1,12-diamine. 1H NMR (500 MHZ, D20) 8 7.97 (s, 2H), 7.89 (d, J = 8.0 HZ, 2H), 7.69-7.66 (m, 3H), 4.43 (s, 4H), 4.37 (s, 2H), 3.14-3.05 (m, 12H), 2.94 (d, J : 7.0, 6H), 2.10-2.01 (m, 3H), 1.78-1.71 (m, 12H), 1.39-1.26 (m, 48H), 1.50 (d, J = 6.5, 18H). 13C NMR (125 MHZ, D20) 8 144.1, 142.5,135.1,133.4,133.2,133.1,132.5,132.1,130.3,130.3, 57.0, 52.9, 52.7, 50.6, 49.4, 49.3, 31.2, 31.1, 31.1, 31.0, 30.6, 28.3, 28.3, 28.2, 28.0, 27.8, 21.7. LRMS [M+H]+ 960.0.

Example 82 Preparation ofNI,N1'-([I,I’-bz‘phenyl]-3,5-a’z‘ylbz‘s(methylene))bis(N4-(3- (hexylamino)propyl)butane-I,4-a’z'amz'ne), hydrochloride salt (CZ-118) CZ: ~H a .3") Example 82 was prepared in a similar fashion to Example 68 (CZ-86) from 5- bromoisophthaldehyde, boronic acid and N1-(3-aminopropyl)-]\73-hexylpropane-1,3- diamine. 1H NMR (500 MHz, D20) 6 7.87 (s, 2H), 7.76 (d, J = 7.5 Hz, 2H), 7.58 (t, J = 8.0 Hz, 3H), 7.51 (t, J = 7.5 Hz, 1H), 4.37 (s, 4H), 3.21-3.14 (m, 12H), 3.07 (t, J = 7.0 Hz, 4H), 2.16-2.09 (m, 4H), 1.90-1.77 (m, 10H), 1.69 (quint, J = 7 Hz, 4H), 1.39-1.37 (m, 4H), 1.32- 1.31(m, 10H), 0.86 (t, J = 7 Hz, 6H). 13C NMR (125 MHz, D20) 5 142.6, 139.1, 132.6, 130.2, 129.7, 129.5, 128.7, 127.3, 50.9, 48.1, 47.2, 46.7, 44.7, 44.5, 30.6, 25.6, 25.5, 23.0, 23.0, 22.8, 21.9, 13.4. LRMS [M+H]+ 637.6.

Example 83 Preparation 0fN1,N1'-((5-(2-ethylbut0xy)—I, 3—phenylene)bis(methylene))bis(IV3-(3- (isobutylammonio)pr0pyl)pr0pane-I,3-dz'aminium) chloride ) Step 1: Dimethyl 5—(2-ethylbut0xy)isophthalate. yl 5-hydroxyisophthalate (0.44 g, 2.10 mmol) cesium carbonate (1.37 g, 4.20 mmol) and CH3CN (20 mL) were added to a round-bottom flask and stirred for 15-30 min. 3-(Bromomethyl)pentane (0.42 g, 2.52 mmol) was added and the reaction was stirred for 16 h. The t was removed under reduced pressure and the reaction mixture was partitioned between EtOAc (50 mL) and H20 (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL). The combined organics were dried over Na2S04, filtered, and concentrated under reduced pressure to afford the desired product which was used without further purification.

Step 2: Ethylbut0xy)-I,3-phenylene)dimethanol. To a solution of dimethyl 5- ylbutoxy)isophthalate (0.62 g, 2.10 mmol) in THF (20 mL) was added LiAlH4 (0.24 g, 6.32 mmol). The reaction mixture was stirred for 8 h and subsequently quenched with 2N HCl (10 mL) and extracted with EtzO (2 x 25 mL) and EtOAc (2 x 25 mL). The combined organics were dried over Nast4, d, and concentrated under reduced pressure to afford the desired product which was used without further purification.

Step 3: 5-(2-Ethylbut0xy)isophthalaldehyde. (5-(2-Ethylbutoxy)—l,3- phenylene)dimethanol (0.50 g, 2.10 mmol) and CH2C12 (10 mL) were added to a round- bottom flask. To this solution was added PCC (1.35 g, 6.30 mmol), and the reaction was stirred for 16 h. The reaction mixture was concentrated under reduced pressure. Purification by column chromatography (90% s/EtOAc) afforded the desired product (0.13 g, 25%, 3 steps) as a white olid. 1H NMR (500 MHz, CDClg) 8 ppm 10.01 (s, 2H), 7.90 (s, 1H), 7.62 (s, 2H), 3.94 (d, J: 5.5 Hz, 2H), 1.68 (hept, J: 6.5 Hz, 1H), 1.46 (dec, J: 6.5 Hz, 4H), 0.91 (t, J = 7.5 Hz, 6H). 13c NMR (125 MHz, CDC13) 5 ppm 191.1, 160.7, 138.5, 124.1, 120.0, 71.1, 40.9, 235,113.

Step 4: N1,Nli-((5-(2-Ethylbut0xy)-I,3-phenylene)bis(methylene))bis(N3-(3— (isobutylamin0)pr0pyl) propane-1,3-diamz'ne). 5-(2-Ethylbutoxy)isophthalaldehyde (0.20 g, 0.88 mmol) and MeOH (15 mL) were added to a round-bottom flask. To the on was added N1-(3-aminopropyl)—]\]3-isobutylpropane-1,3-diamine (0.33 g, 1.76 mmol) and the reaction mixture was stirred for 24 h. Sodium borohydride (0.13 g, 3.52 mmol) was added and the reaction mixture d for l h. The reaction mixture was concentrated under d pressure to afford a white solid. Aqueous NaOH (10%, 50 mL) and EtOAc (50 mL) were added and the reaction mixture stirred for l h. The layers were separated and the aqueous layer was extracted with EtOAc (50 mL). The organic layers were combined, dried over NaZSO4, filtered, and concentrated under reduced pressure to afford the desired product a clear 011.

Step 5: N1,Nli—((5—(2—Ethylbut0xy)—I,3—phenylene)bis(methylene))bis(N3—(3— (isobutylammonio) propprropane-I,3-dz'amim'um) chloride. To N1,Nly-((5-(2-ethylbutoxy)- 1,3-phenylene)bis(methylene))bis(]\73-(3-(isobutylamino)propyl) propane-1,3-diamine) from step 4 was which was added methanolic HCl (50 mL, 1.0M). The reaction mixture was d for 1 h, the reaction e was concentrated under reduced pressure and the solid collected by vacuum filtration. The solid was washed with EtZO (10 mL) and hot MeOH (10 mL) to afford the desired product (0.42 g, 59%) as a white solid. 1H NMR (500 MHz, D20) 8 7.20 (s, 2H), 7.18 (s, 1H), 4.29 (s, 4H), 4.05 (d, J = 6.0 Hz, 2H), 3.25-3.15 (m, 16H), 2.94 (d, J = 7.0 Hz, 2H), 2.21-2.12 (m, 8H), 2.07-1.99 (m, 2H), 1.72 (p, J = 6.5 Hz, 1H), 1.50-1.43 (m, 4H), 1.00 (d, J = 7.0 Hz, 12H), 0.92 (t, J = 6.0 Hz, 6H). 13C NMR (125 MHz, D20) 8 159.6, 132.9, 123.4, 117.4, 71.4, 54.8, 50.8, 44.7, 44.6, 44.6, 44.1, 40.1, 25.5, 22.6, 22.5, 19.0, 10.2. LRMS [M+H]+577.6.

Example 84 Preparation ofNI,N1’—((5-isopropoxy-l,3-phenylene)bis(methylene))bis(N3-(3- aminoproprpropane-I,3-diamine), hydrochloride salt (CZ-93) < NH2 l"N_\_/NH 2.9 2 Example 84 was prepared in a similar n to Example 83 (CZ—91) from 5- isopropoxyisophthalaldehyde and tert—butyl (3-((3-aminopropy1)amino)propyl)carbamate. 1H NMR (300 MHZ, D20) 8 7.13 (s, 3H), 4.78-4.72 (m, 1H), 4.23 (s, 4H), 3.27-3.18 (m, 12H), 3.13 (t, J = 7.8 HZ, 4H), 2.23—2.18 (m, 4H), 2.18—2.10(m, 4H), 1.36 (d, J =10.2HZ, 6H).

Example 85 Preparation I ’—((5-(cyclohexylmethoxy)-I, 3-phenylene)bis(methylene))bis(N3-(3- (octylamino)propyl)propane-1,3-diamine), hydrochloride salt (CZ-103) .1144?” ..29 H;

[0524] Example 85 was prepared in a similar fashion to Example 83 (CZ-91) from dimethyl 5-butoxyisophtha1ate, (bromomethyl)cyclohexane, tert-butyl (3 -((3- aminopropyl)amino)propyl)carbamate and octanal. 1H NMR (500 MHZ, D20) 5 7.18 (s, 3H), 4.28 (s, 4H), 3.95 (s, 2H) .14 (m, 16H), 3.06 (t, J = 7.5 HZ, 4H), .10 (m, 8H), .80 (m, 3H), 1.76-1.64 (m, 8H), 1.41-1.20 (m, 22H), 1.12-1.03 (m, 2H), 0.89-0.83 (m, 6H). 13C NMR (125 MHZ, D20)8159.5,133.0, 123.4, 117.3, 74.4, 50.8, 47.8, 44.6, 44.6, 44.2, 44.1, 36.8, 30.9, 29.1, 28.1, 28.0, 25.9, 25.6, 25.4, 25.2, 22.6, 22.6, 21.9, 13.3. LRMS [M+H]+701.7.

Example 86 ation ofNI,N1’—((5-(cyclohexyloxy)-1,3-phenylene)bis(methylene))bis(N3-(3- (isobutylamino)propyl)propane-1,3-diamine), hydrochloride salt (CZ-104) I"'\'_\_/ Example 86 was prepared in a similar fashion to Example 83 (CZ-91) from 5- (cyclohexyloxy)isophthalaldehyde and N1-(3-aminopropyl)-N3-isobutylpropane-1,3-diamine. 1H NMR (300 MHZ, D20) 5 7.60 (s, 2H), 7.53 (s, 1H), 4.73 (t, J = 1.2 Hz, 1H), 4.42 (s, 4H), 3.26-3.13 (m, 16H), 2.84 (d, J = 6.9 Hz, 2H), .08 (m, 8H), 2.06-1.96 (m, 2H), 1.87-1.78 (m, 4H), 1.65-1.36 (m, 6H), 0.99 (d, J : 6.6 Hz, 12H). LRMS [M+H]+ 575.5.

Example 87 ation ofNI,N1V—((5-(cyclohexyloxy)-1,3-phenylene)bis(methylene))bis(N3-(3- aminopropyl)propane-1,3-diamine), hydrochloride salt (CZ-105) < NH2 HN—\_/NH ‘23 Example 87 was prepared in a similar fashion to Example 83 (CZ-91) from 5- (cyclohexyloxy)isophthalaldehyde and tert-butyl (3-((3-am1nopr0pyl)amino)propyl)- carbamate. 1H NMR (300 MHz, D20) 5 7.22 (s, 2H), 7.18 (s, 1H), 4.68 (t, J = 1.2 Hz, 1H), 4.30 (s, 4H), 3.28-3.18 (m, 12H), 3.14 (t, J = 7.8 Hz, 4H), 2.21-2.06 (m, 8H), 1.84-1.75 (m, 4H), 1.63-1.34 (m, 6H). LRMS [M+H]+ 463.5.

Example 88 Preparation 1V—((5-(benzyloxy)—I,3-phenylene)bis(methylene))bis(N3-(3- (isobutylamino)propyl)propane-I,3-diamine), hydrochloride salt (CZ-106) “EQU— Qi W3 $7 - 6 HCI

[0527] Example 88 was prepared in a similar fashion to Example 83 (CZ-91) from dimethyl 5-butoxyisophthalate, benzyl bromide, utyl (3-((3-aminopropyl)amino)- propyl)carbamate and isobutyraldehyde.

Example 89 Preparation 1V—(I,3—phenylenebis(methylene))bis(N3—(3—((Cyclohexylmethyl)— amino)propyl)propane-1,3-diamine), hydrochloride salt (CZ-97) < HN HN_\_/NH Step 1: -(1,3-Phenylenebis(methylene))bis(1V3-(3-((Cyclohexylmethyl)- amino)propyl)propane-1,3-diamine). Isophthalaldehyde (0.34 g, 2.57 mmol, l) and MeOH (20 mL) were added to a round-bottom flask. To the solution was added N1-(3 -aminopropyl)- N3-(cyclohexylmethyl)propane-1,3-diaminediamine (1.17 g, 5.14 mmol, 2) and the reaction mixture was stirred at rt for 24 h. Sodium borohydride (0.39 g, 10.28 mmol, 4) was added and the on mixture stirred for 1 h. The reaction mixture was concentrated under reduced pressure to afford a white solid. Aqueous NaOH (10%, 100 mL) and EtOAc (100 mL) were added and the reaction mixture stirred for l h. The layers were separated and the aqueous layer was extracted with EtOAc (100 mL). The organic layers were combined, dried over Na2S04, filtered, and trated under reduced pressure to afford a clear oil which was used t filI‘thGI‘ purification.

Step 2: N1,NI'-(I,3-phenylenebis(methylene))bis(N3—(3-((cyclohexylmethyl)amin0)- propyl)propane-1,3-diamine) hydrochloride salt. To the crude N1,N1‘-(1,3-phenylenebis- (methylene))bis(l\73-(3-((cyclohexylmethyl)amino)propyl)propane-1,3 -diamine) from Step 1 was added olic HCl (100 mL, 1.0M). The reaction mixture stirred at rt for 2 h. The reaction mixture was concentrated under reduced pressure and the solid was collected by vacuum filtration and washed with EtZO (50 mL) and hot MeOH (50 mL) to afford the desire t (1.3 g, 67%) as a white solid. 1H NMR (500 MHz, D20) 8 7.57 (s, 4H), 4.30 (s, 4H), 3.22 (t, J = 7.5 Hz, 4H), 3.18-3.15 (m, 8H), 3.12 (t, J = 8.0 Hz, 4H), 2.91 (d, J = 7.0 Hz, 4H), 2.18-2.08 (m, 8H), 1.72-1.62 (m, 12H), 1.28-1.11 (m, 6H), 1.03-0.96 (m, 4H). 13C NMR (125 MHz, D20) 8 131.4, 131.2, 131.0, 130.2, 53.7, 50.8, 48.8, 44.7, 44.6, 44.0, 34.5, 29.7, .3, 24.8, 22.6, 22.5. LRMS [M+H]+ 557.5. e 90 Preparation ofNI,NI V—(I, 3-phenylenebis(methylene))bis(N3-(3-(octylamino)propyl)propane- 1,3-diamine), hydrochloride salt (CZ-100) < HN m. HN—\_/NHHN -6HC| Example 90 was prepared in a similar fashion to Example 62 (CZ-76) from isophthaldehyde, tert—butyl (3 —((3—aminopropyl)amino)propyl)carbamate and octanal. 1H NMR (500 MHZ, D20) 8 7.58 (s, 4H), 4.32 (s, 4H), 3.10-2.98 (m, 16H), 2.91 (t, J = 8.0 Hz, 4H), 2.05-1.95 (m, 8H), 1.53 (p, J = 7 Hz, 4H), 1.24-1.13 (m, 20H), 0.71 (t, J = 7.0 Hz, 6H). 13C NMR (125 MHz, D20)5131.4, 131.2, 131.1, 130.2, 50.8, 47.9, 44.6, 44.6, 44.2, 44.1, .9, 28.1, 28.1, 25.6, 25.4, 22.6, 219,133. LRMS [M+H]+ 589.5.

Example 91 Preparation ’1(1,3—phenyzenebz‘smethylene»[mar—(3— (isobutylamino)propyl)propane-I,3-diamine), hydrochloride salt 8) HNMQNWF >_/ '6HCI

[0531] Example 91 was prepared in a similar fashion to Example 89 (CZ-97) from isophthaldehyde and N1-(3-aminopropyl)—]\73-isobutylpropane—1,3-diamine. 1H NMR (300 MHz, D20) 6 7.54 (s, 4H), 4.28 (s, 4H), 3.19-3.08 (m, 16H), 2.88 (d, J= 7.5 Hz, 4H), 2.15- 2.05 (m, 8H), 2.02-1.90 (m, 2H), 0.95 (d, J: 6.9 Hz, 12H). LRMS [M+H]+ 477.4.

Example 92 Preparation ofNI,N1V—((5-bromo-I,3-phenylene)bis(methylene))bis(N3-(3- (isobutylamino)propyl)propane-I,3-diamine), hydrochloride salt (CZ-109) HNLQN} >_/ -6HC| Example 92 was prepared in a similar fashion to Example 89 (CZ-97) from 4- bromo-isophthaldehyde and aminopropyl)—N3-isobutylpropane-1,3-diamine. 1H NMR (300 MHz, D20) 8 7.76 (s, 2H), 7.51 (s, 1H), 4.27 (s, 4H), 3.21-3.08 (m, 16H), 2.88 (d, J: 7.5 Hz, 4H), 2.16-2.06 (m, 8H), 2.02-1.93 (m, 2H), 0.96 (d, J: 6.6 Hz, 12H). LRMS [M+H]+ 555.4, 557.4.

Example 93 Preparation ofNI,NI V—(I, 3-phenylenebis(methylenejjbis(N3-(3-(hexylaminojpropyljpropane- 1,3-diamine), hydrochloride salt (CZ-113) (—1111 HN_\_/NH -6HCI

[0533] Example 93 was prepared in a similar fashion to Example 89 (CZ-97) from haldehyde and N1-(3-aminopropyl)—l\73-hexylpropane-1,3-diamine. 1H NMR (500 MHZ, D20) 8 7.58 (s, 3H), 4.32 (s, 4H), .13 (m, 16H), 3.05 (t, J = 8.0 Hz, 4H), 2.20- 2.09 (m, 8H), 1.68—1.63 (m, 4H), 1.38—1.33 (m, 4H), 1.32—1.26 (m, 8H), 0.85 (t, J = 7.5 Hz, 6H). 13C NMR (125 MHz, D20) 8 131.6, 131.5, 131.4, 130.4, 51.1, 48.1, 44.9, 44.8, 44.5, 44.3, 30.6, 25.6, 25.5, 22.8, 21.9, 13.5. LRMS [M+H]+533.6. e 94 Preparation ofNI,N1'-(1,3-phenylenebis(methylene))bis(N4-(3-(hexylamino)propyl)butane- 1,4-diamine), hydrochloride salt (CZ-119) g HN—\_/—Nl-l—/—NH HN§ - 6 HCI

[0534] Example 94 was prepared in a similar fashion to Example 89 (CZ-97) from haldehyde and N1-(3-aminopropyl)—N4-hexylbutane-1,4-diamine. 1H NMR (500 MHz, D20) 6 7.65 (s, 4H), 4.37 (s, 4H), 3.26-3.19 (m, 16H), 3.14 (t, J= 7.5 Hz, 4H), 2.20 (quint, J = 8.0 Hz, 4H), 1.88 (br s, 8H), 1.75 (quint, J: 8.0 Hz, 4H), 1.46-1.42 (m, 4H), 1.39-1.36 (m, 8H), 0.94 (t, J: 6.5 Hz, 6H).13C NMR (125 MHz, D20) 8 134.2, 133.7, 133.5, 132.7, 53.3, 50.4, 49.6, 49.1, 47.1, 46.8, 32.9, 27.9, 27.8, 25.4, 25.3, 25.2, 24.2, 15.8. LRMS [M+H]+ 561.5.

Example 95 Preparation 1',N1”-([1, I '-bz'phenyl]-3,3 iyltris(methylene))tris(N3-(3— (isobutylamino)pr0pyl)propane-1 3-a’z‘amine) hydrochloride salt (CZ-99) New;Eng YNMNMNQ - 9 HCIH H H Step 1: Bz'phenyl]—3,3Z5—tricarbaldehyde. A solution of 5— bromoisophthaldehyde (2.13 g, 10.0 mmol), 3-formylphenylboronic acid (1.49 g, 10.0 mmol) and potassium carbonate (2.76 g, 20.0 mmol) in DME/HZO (5:1 50 mL) was purged with N2 for 5 min. Tetrakispalladium triphenylphosphine (0.12 g, 0.1 mmol) was added and the on mixture was heated to 90 CC and stirred for 16 h. The on e was cooled, filtered through a pad of Ce1ite diatomaceous earth and the solvent evaporated under reduced pressure. Purification by column chromatography (10% EtOAc/hexanes) afforded the desired product (1.47 g, 62%) as a tan solid. 1H NMR (300 MHz, CDClg) 6 ppm 10.20 (s, 2H), 10.10 (s, 1H), 8.42 (s, 3H), 8.21 (s, 1H), 7.98 (t, J: 5.4 Hz, 2H), 7.72 (t, J: 5.4 Hz, 2H).

Step 2; N’,N”,N’ ”_([1,1 '—Biphenyl]—3,3 ',5—trz‘yltrz‘s (methylene))ms (N3—(3— (isobutylamino)propyl)propane-1,3-a’z'amz'ne). [1,1'—biphenyl]—3,3',5-tricarbaldehyde (0.24 g, 1.0 mmol) and MeOH (20 mL) were added to a round-bottom flask. To the solution was added N1—(3—aminopropyl)—N3—isobutylpropane—1,3—diamine (570 mg, 3.0 mol, 3 equiv.) and the reaction mixture was stirred at rt for 24 h. Sodium borohydride (0.34 g, 9.0 mmol) was added and the reaction mixture stirred for 1 h. The reaction mixture was concentrated under reduced pressure to afford a white solid. Aqueous NaOH (10%, 100 mL) and EtOAc (100 mL) were added and the reaction mixture stirred for 1 h. The layers were separated and the aqueous layer was extracted with EtOAc (100 mL). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to afford a clear oil which was used t r purification.

Step 3: ,N1H-([I,I'-Biphenyl]-3,3',5-triyltris(methylene))trisW3-(3- (isobutylamino)propyl)propane-I,3-diamine), hydrochloride salt. N1,N1',N1"-([1,1'- biphenyl]-3 ,3',5 -triyltris(methylene))tris(N3-(3-(isobutylamino)propyl)propane-1 ,3-diamine) from step 2 was subjected to acidification with methanolic HCl (100 mL, 1.0M). The reaction mixture d at rt for 2 h. The reaction mixture was concentrated under reduced re and the solid was collected by vacuum filtration and washed with Et20 (50 mL) and hot MeOH (50 mL) to afford the desired product (0.58 g, 52%) as a white solid. 1H NMR (500 MHz, D20) 8 ppm 7.91 (s, 1H), 7.85-7.82 (m, 3H), 7.65-7.56 (m, 3H), 4.42 (s, 4H), 4.39 (s, 2H), 3.29-3.16 (m, 24H), 2.93 (d, J = 6.0 Hz, 6H), 2.20-2.13 (m, 12H), 2.07-2.00 (m, 3H), 0.99 (d, J : 6.0 Hz, 18H). 13CNMR (125 MHz, D20) 8 ppm 141.6, 139.8, 132.2, 131.3, 130.4, 130.3, 129.7, 129.5, 128.6, 128.4, 54.8, 54.8, 51.1, 50.8, 44.7, 44.6, 44.2, 44.0, 25.5, 22.7, 22.5, 19.0. LRMS [M+2H+]2+ 376.8.

Example 96 Preparation ofNI,NI '—((5-(3-(isoButylammonio)propoxy)-1, 3-phenylene)bis(methylene))— bis(N3-(3-ammoniopropyl)propane-1,3-diaminium) chloride (CZ-107) ( NH2 |"'\'—\_/NH H 2 0\—\—th Step 1: tert-Butyl (3-hydroxypropyl) (isobutyl)carbamate. 3-(iso- Butylamino)propan—1—ol (7.92 g, 60.5 mmol) and THF (100 mL) were added to a round— bottom flask. To the solution was added aqueous NaOH (10%, 100 mL) followed by the slow addition of di-tert-butyl dicarbonate (11.86 g, 54.4 mmol). The reaction mixture was stirred for 12 h. The reaction mixture was extracted with CHzClz (3 x 100 mL). The combined organics were dried over Na2SO4, filtered, and concentrated under reduced pressure to afford the desired product (12.0 g, 86%) which was used without further purification.

Step 2: 3-((tert—But0xycarb0nyl) (isobutybamz‘nomropyl methanesulfonate. tert- Butyl (3-hydroxypropyl)(isobutyl)carbamate (0.73 g, 3.15 ), triethylamine (0.64 g, 6.30 mmol) and CH2C12 (30 mL) were added to a round-bottom flask. To the solution was added methanesulfonyl de (0.54 g, 4.72 mmol) and the reaction mixture was stirred for 5 h.

The reaction mixture was concentrated under d pressure and purified by column chromatography (hexanes/EtOAC) to afford the desired product (063 g, 64%) as a yellow oil.

Step 3: Dimethyl 5-(3-((tert—but0xycarb0nyl)(isobutyl)amz'n0)pr0p0xy)is0phthalate.

Dimethyl 5-hydroxyisophthalate (0.30 g, 1.44 mmol) cesium carbonate (0.95 g, 2.88 mmol) and CH3CN (25 mL) were stirred for 30 min. 3-((tert—Butoxycarbonyl)(isobutyl)amino)- propyl methanesulfonate (0.63 g, 2.02 mmol) was added and the reaction mixture was stirred for 16 h. The on mixture was concentrated under reduced pressure and ioned between EtOAc (50 mL) and H20 (50 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL). The combined organics were dried over NazSO4, filtered, and concentrated under reduced pressure to afford the desired product, which was used t further purification.

Step 4: tert—Butyl (3-(3,5-bis(hydroxymethyl)phen0xy)pr0pyl)(isobutybcarbamate.

To a solution of dimethyl 5-(3-((tert-butoxycarbonyl)(isobutyl)amino)propoxy)isophthalate (0.60 g, 1.44 mmol) in THF (10 mL) was added LiAlH4 (0.30 g, 7.89 mmol). The reaction mixture was stirred for 8 h and uently quenched with 2N HCl (10 mL) and extracted with Et20 (2 x 25 mL) and EtOAc (2 x 25 mL). The combined organic layers were dried over N32804, filtered, and concentrated under reduced re to afford the desired product (0.28 g, 54%) as an oil, which was used without further purification.

Step 5: tert—Butyl (3-(3,5-dl'f0rmylphen0xy)pr0pyl) (isobutyl)carbamate. tert—Butyl (3-(3,5-bis(hydroxymethyl)phenoxy)propyl)(isobutyl)carbamate (0.29 g, 0.78 mmol) and CH2C12 (20 mL) were added to a round—bottom flask. To the solution was added PCC (0.42 g, 1.94 mmol) and the reaction was stirred for 16 h. The reaction e was concentrated under reduced pressure. Purification by column chromatography (90% Hexanes/EtOAc) afforded the desired t (0.15 g, 20% 3 steps) as a white semi—solid, which was used without further purification.

Step 6: N1,N1/-((5-(3-(iso-Butylaminwpropoxy}I,3-phenylene)bis(methylene))- bis(N3-(3-amin0pr0pyl)pr0pane-1,3-a’z'amz'ne). utyl (3-(3,5-diformylphenoxy)propyl)- (isobutyl)carbamate (0.15 g, 0.42 mmol) and MeOH (10 mL) were added to a round-bottom flask. To the on was added tert-butyl (3-((3-aminopropyl)amino)propyl)carbamate (0.20 g, 0.84 mmol) and the reaction mixture was d for 24 h. Sodium borohydride (0.06 g, 1.69 mmol) was added and the reaction mixture d for 1 h. The reaction mixture was concentrated under reduced pressure to afford a white solid. Aqueous NaOH (10%, 50 mL) and EtOAc (50 mL) were added, the layers separated and the aqueous layer was extracted with EtOAc (50 mL). The organic layers were combined, dried over NaZSO4, filtered, and concentrated under reduced pressure to afford the desired product as clear oil which was used without further purification.

[0544] Step 7: N1,N1/-((5-(3-(iso-Batylammonio)propoxy)-I,3-phenylene)bis(methylene))- bis(N3-(3-ammoniopropyl)propane-1,3-diaminiam) chloride. To N1,N1'-((5-(3-(iso- butylamino)propoxy)—1 ,3 -phenylene)bis(methylene))bis(l\73-(3 -an11nopropyl)propane- 1 ,3 - diamine) from step 6 was added methanolic HCl (50 mL, 1.0M). The reaction mixture was stirred for 1 h, the reaction mixture was concentrated under reduced pressure and the solid collected by vacuum filtration. The solid was washed with EtZO (10 mL) and hot MeOH (10 mL) to afford the desired product (0.14 g, 43%) as a white solid. 1H NMR (500 MHz, D20) 8 ppm 7.21 (s, 1H), 7.19 (s, 2H), 4.30 (s, 4H), 4.23 (t, J: 5.5 Hz, 2H), 3.29 (t, J: 7 Hz, 2H), .19(m, 12H), 3.12 (t, J: 7.5 Hz, 4H), 2.95 (d, J: 7 Hz, 2H), 2.25—2.03 (m, 11H), 1.01 (d, J= 7.5 Hz, 6H). 13c NMR (125 MHz, D20) 158.9, 133.0, 123.7, 117.1, 65.8, 54.8, 50.7, 45.7, 44.7, 44.6, 44.2, 36.7, 25.5, 25.2, 23.7, 22.6, 19.1. LRMS [M+H]+ 494.4.

Example 97 Preparation 1'—((5-(3-(isobutylamino)propoxy)-I,3-phenylene)bis(methylene))bis(N3- (3-(octylamino)propyl)propane—I,3-diamine), hydrochloride salt (CZ-110) (711.

HN—\_/NH Step 1: tert-Butyl (3-hydr0xypr0pyl) (isobutylkarbamate. 3-(l's0- Butylamino)propanol (7.92 g, 60.5 mmol) and THF (100 mL) were added to a round- bottom flask. To the solution was added aqueous NaOH (10%, 100 mL) followed by the slow addition of di-tert-butyl dicarbonate (11.86 g, 54.4 mmol). The reaction e was stirred for 12 h. The reaction mixture was extracted with CHZClz (3 x 100 mL). The combined cs were dried over NagSO4, filtered, and concentrated under reduced pressure to afford the desired product (12.0 g, 86%), which was used without further purification.

Step 2: 3-((tert—But0xycarb0nyl) nzl)amin0)pr0pyl esulfonate. tert- Butyl (3-hydroxypropyl)(isobutyl)carbamate (0.73 g, 3.15 ), triethylamine (0.64 g, 6.30 mmol) and CH2C12 (30 mL) were added to a round-bottom flask. To the solution was added methanesulfonyl chloride (0.54 g, 4.72 mmol) and the reaction mixture was stirred for 5 h.

The reaction mixture was trated under reduced pressure and purified by column tography (hexanes/EtOAC) to afford the desired product (0.63 g, 64%) as a yellow oil.

Step 3: Dimethyl 5-(3-((tert—but0xycarb0nyl)(isobutyl)amin0)pr0p0xy)is0phthalate: Dimethyl 5-hydroxyisophthalate (0.30 g, 1.44 mmol) cesium carbonate (0.95 g, 2.88 mmol) and CH3CN (25 mL) were stirred for 30 min. 3-((tert-Butoxycarbonyl)(isobutyl)amino)- propyl methanesulfonate (0.63 g, 2.02 mmol) was added, and the reaction mixture was stirred for 16 h. The reaction mixture was concentrated under reduced pressure and partitioned between EtOAc (50 mL) and H20 (50 mL). The layers were separated, and the aqueous layer was extracted with EtOAc (50 mL). The ed organics were dried over NazSO4, filtered, and concentrated under reduced pressure to afford the desired product, which was used without further purification.

[0548] Step 4: tert-Butyl (3-(3,5-bis(hydroxymethpr/aenoxy)pr0pyl)(isobugibcarbamate.

To a on of dimethyl 5-(3-((tert-butoxycarbonyl)(isobutyl)amino)propoxy)isophthalate (0.60 g, 1.44 mmol) in THF (10 mL) was added LiAlH4 (0.30 g, 7.89 mmol). The reaction mixture was stirred for 8 h and subsequently quenched with 2N HCl (10 mL) and ted with EtZO (2 x 25 mL) and EtOAc (2 x 25 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to afford the desired t (0.28 g, 54%) as an oil, which was used without further purification.

Step 5: tert-Butyl (3-(3,5-dz‘f0rmylphenoxy)pr0pyl) (isobutyDCCIrbamate. tert-Butyl (3-(3,5-bis(hydroxymethyl)phenoxy)propyl)(isobutyl)carbamate (0.29 g, 0.78 mmol) and CH2C12 (20 mL) were added to a bottom flask. To the solution was added PCC (0.42 g, 1.94 mmol) and the reaction was stirred for 16 h. The on mixture was concentrated under reduced pressure. Purification by column chromatography (90% hexanes/EtOAc) afforded the desired product (0.15 g, 20% 3 steps) as a white semi-solid which was used without further purification.

Step 6: NI,NI/-((5-(3-(iso-Butylaminwpropoxy}I,3-phenylene)bis(methylene))- bis(N3- (3-amz'n0pr0py0pr0pane-1,3-dz'amz'ne). tert-Butyl (3-(3,5-diformylphenoxy)propyl)- (isobutyl)carbamate (0.15 g, 0.42 mmol) and MeOH (10 mL) were added to a round-bottom flask. To the solution was added tert—butyl (3-((3-aminopropyl)amino)propyl)carbamate (0.20 g, 0.84 mmol), and the reaction mixture was stirred for 24 h. Sodium borohydride (0.06 g, 1.69 mmol) was added and the reaction mixture d for 1 h. The reaction mixture was concentrated under reduced re to afford a white solid. Aqueous NaOH (10%, 50 mL) and EtOAc (50 mL) were added, the layers separated, and the aqueous layer was extracted with EtOAc (50 mL). The organic layers were combined, dried over Na2S04, filtered, and concentrated under reduced pressure to afford the desired product as a clear oil, which was used without further purification.

Step 7: NI,NI/-((5-(3-(iso-Butylamm0ni0)pr0p0xy)-1,3- phenylene)bis(methylene))bis(N3-(3-amm0ni0pr0pyl)pr0pane-I,3-dz'aminium) chloride. To N1,N1'-((5 -(3 -(z's0-Butylamino)propoxy)-1,3 -phenylene)bis(methylene))bis(N3-(3 - aminopropyl)propane—l,3—diamine) from step 6 was added methanolic HCl (50 mL, 1.0M).

The reaction mixture was stirred for 1 h, the reaction e was concentrated under reduced pressure and the solid collected by vacuum filtration. The solid was washed with EtzO (10 mL) and hot MeOH (10 mL) to afford the desired t (0.14 g, 43%) as a white solid. 1H NMR (500 MHz, D20) 6 ppm 7.21 (s, 1H), 7.19 (s, 2H), 4.30 (s, 4H), 4.23 (t, J: 5.5 Hz, 2H), 3.29 (t, J: 7 Hz, 2H), 3.25-3.19 (m, 12H), 3.12 (t, J: 7.5 Hz, 4H), 2.95 (d, J: 7 Hz, 2H), 2.25—2.03 (m, 11H), 1.01 (d, J: 7.5 Hz, 6H). 13C NMR (125 MHZ, D20) 158.9, 133.0, 123.7, 117.1, 65.8, 54.8, 50.7, 45.7, 44.7, 44.6, 44.2, 36.7, 25.5, 25.2, 23.7, 22.6, 19.1. LRMS [M+H]+494.4.

Step 8: To N1,N1'-((5-(3-(isoButylammonio)propoxy)-1,3-phenylene)bis- lene))bis(]\73-(3 -ammoniopropyl)propane-1,3-diaminium) hydrochloride salt was added aqueous NaOH (10%, 100 mL) and 75% CHClg/isopropanol (100 mL). The layers were separated, and the aqueous layer was extracted with 75% CHClg/isopropanol (4 x 100 mL). The organic layers were combined, dried over Na2SO4, d, and concentrated under reduced pressure to afford a clear oil, which was used without r purification.

Step 9: To a round-bottom flask was added the crude N1,N1'-((5-(3- (isobutylammonio)propoxy)- 1 ,3 -phenylene)bis(methylene))bis(l\l3-(3 - ammoniopropyl)propane-1,3-diaminium) (0.91 g, 1.84 mmol) and MeOH (20 mL). To the solution was added octanal (0.47 g, 3.68 mmol) and the reaction e was stirred at rt for 24 h. Sodium borohydride (0.28 g, 7.36 mmol) was added and the reaction mixture was stirred for 1 h. The reaction mixture was concentrated under reduced re to afford a white solid. Aq. NaOH (10%, 100 mL) and EtOAc (100 mL) were added and the mixture d for 1 h. The layers were separated and the aqueous layer was extracted with EtOAc (100 mL). The combined organics were dried over Na2804, filtered, and concentrated under reduced re to afford the desired product as a clear oil which was used without further ation.

Step 10: N1,N1'-((5-(3-(isobatylamino)propoxy)-I,3-plzenylene)bis(methylene))— bis(N3-(3-(octylamino)propyl)propane-I,3—diamine), hydrochloride salt. To the crude N1,N1'- ((5 -(3 -(isobutylamino)propoxy)-1,3 -phenylene)bis(methylene))bis(N3-(3 - (octylamino)propyl)propane-1,3-diamine) was added methanolic HCl (100 mL, 1.0M). The reaction mixture was stirred at rt for 1 h. The reaction mixture was concentrated under reduced pressure, and the solid was ted by vacuum filtration and washed with Et20 (50 mL) and hot MeOH (50 mL) to afford the desired product as a white solid (0.75 g, 42%). 1H NMR (500 MHz, D20) 5 7.21 (s, 1H), 7.20 (s 2H), 4.30 (s, 4H), 4.24 (t, J = 5 Hz, 2H), 3.30 (t, J = 7 Hz, 2H), 3.26-3.14 (m, 16H), 3.07 (t, J = 7 Hz, 4H), 2.96 (d, J = 7.5 Hz, 2H), 2.26- 2.03 (m, 11H), 1.69 (p, J = 6.5 Hz, 4H), 1.38—1.28 (m, 20H), 1.02 (d, J = 6.5 Hz, 6H), 0.87 (t, J = 5.5 Hz, 6H). 13C NMR (125 MHZ,D20)8158.9,133.0,123.7, 117.1, 65.8, 54.8, 50.8, 47.9, 45.7, 44.6, 44.6, 44.2, 44.1, 30.9, 28.1, 28.1, 25.6, 25.5, 25.4, 25.2, 22.6, 22.6, 19.1, 13.4.

Example 98 Antimicrobial activity ofpolyamines in agar media A 0.5 McFarland standard of each bacteria was made, then individual lawns of bacteria spread onto the surface of tryptic soy agar (TSA). 500 uL of the suspended gel were placed on the center of each lawn. Plates were incubated at 37 C for 24 hours, then imaged.

Images of the resulting gels are shown in FIGS. 30A to 301 show the effects of the polyamines CZ-86, CZ-100, and CZ-110 on various bacterial cell cultures. The data shown indicated that when the polyamine compounds were mixed with the gel, the compounds diffused out of the gel and created zones of inhibition in the bacterial lawns. The gel alone showed no signs of crobial activity (B).

The hydrogel that was used for those images is a LifeCore gel with 1% sodium hyaluronate (1% Sodium Hyaluronate Solution Part #82). The CZ compound was suspended in water at 1% (w/w) concentration and ed with the LifeCore gel 1:1 for a final product that contained 0.5% sodium hyaluronate and 0.5% CZ compound.

Example 99 CZ-86 and CZ-90p01yaml'ne hemolysz's activity The polyamines CZ—86 and CZ—90 were tested for hemolytic ty. The difference between the hemolytic indexes of both polyamines and the negative l was 0.00 percent. This places both test es in the non-hemolytic range according to the grade outlined below in Table 6A.

Table 6A: Hemol ic Index and Grade: Hemol ic Index Hemol tic Grade 1 Hemol ic All test method acceptance criteria were met. The test procedures listed below were followed without deviation.

Table 6B: Results Test Article / Optical Average tic Average Corrected Control Density Optical Index Hemolytic Hemolytic Density Index (% Index (% Hemolysis) Hemolysis) Negative Control Test Article / Average Hemolytic e Corrected Control l Index Hemolytic Hemolytic Density Index (% Index (% Hemolysis) sis) Positive 97.068 101.7 Control 105.638 102.255 Phosphate Buffered Saline (PBS) Blank Table 6C: Hemo_lobin Standard: Reression Outut nt 0.00056 Standard Error ofY Estimate 0.00604 R2 0.99961 De _rees of Freedom X Coefficient s 1.44547 Standard Error of Coefficient 0.01164 Acceptance Criteria: The negative control must produce a ted hemolytic index of less than 2%. The positive control must produce a corrected hemolytic index of greater than 5%.

Procedure: The PBS used in testing was calcium and magnesium free. The method has been validated using human blood from one donor. This is in compliance with ISO 10993-4 which states due to differences in blood activity, human blood should be used where possible. Furthermore, pooling unmatched human blood from multiple donors may cause red blood cell agglutination which in turn could cause hemolysis; this supports the use of a single donor.

The blood was drawn using vacutainers containing 0.1 M sodium citrate at a ratio of 9:1 (3.2% anticoagulant to blood). The blood in this test was used within four hours ofblood draw. The ted blood was refrigerated until g was performed.

A hemoglobin standard was d with Drabkin's reagent to give solutions at concentrations of 0.80, 0.60, 0.40, 0.30, 0.20, 0.10, 0.02, and 0.01 mg/ml. These solutions were allowed to stand at room temperature for a minimum of five minutes. The absorbance was read on a spectrophotometer at 540 nanometers (nm). A standard curve was determined with the absorbance values and the standard concentrations of hemoglobin.

Human blood was centrifuged at 700-800 x g for 15 minutes. A 1 mL aliquot of the plasma was added into 1 mL of n's reagent, and placed at room temperature for a minimum of 15 minutes. The absorbance was read on a ophotometer at 540 nm. The hemoglobin concentration was determined from the standard curve and then multiplied by a factor of 2 to obtain the total plasma free hemoglobin. The plasma free hemoglobin was less than 2 mg/mL (actual value 0.221 mg/mL). A 20 uL aliquot of blood was added to 5 mL of Drabkin's reagent in duplicate and allowed to stand at room temperature for a minimum of 15 minutes. The absorbance was read on a spectrophotometer at 540 nm then multiplied by 251 to account for the dilution.

[0564] Based on the plasma hemoglobin and the blood absorbance against Drabkin's reagent, the blood was diluted out to 10 :: 1 mg/mL with PBS. To verify the blood on, a 300 [LL t of this blood was added to 4.5 mL of Drabkin's reagent in cate and allowed to stand at room temperature for a minimum of 15 minutes. The absorbance was read on a spectrophotometer at 540 nm then multiplied by 16 to account for the dilution.

[0565] Glass test tubes were labeled appropriately. Both samples were prepared the same way. The samples were allowed to thaw prior to testing. To each test tube, 7 mL of the test article and 1 mL of the diluted blood were added. The controls consisted of the appropriate amount of control material, 7 mL of PBS and 1 mL of blood. Three tubes were prepared for each test article and control. The tubes were ted at 37 :: 2°C for 3 hours :: 5 minutes.

Tubes were gently inverted twice at 30 minute intervals throughout the incubation period. A non-hemolytic ve control, a hemolytic ve control and a PBS blank were included.

After incubation, test articles were centrifuged at 700-800 K g for 15 minutes and 1 mL of the supernatant fluid was ed with 1 mL of Drabkin's reagent and allowed to stand at room temperature for a minimum of 15 minutes. Following the fugation phase, the supernatant of both test articles visually appeared clear and were particulate free. The PBS blank and the negative control supernatant visually appeared clear and were particulate free. The supernatant of the positive control visually appeared red and were particulate free.

The test articles and controls were read at 540 nm in a spectrophotometer.

The hemolytic index (percent hemolysis) was interpreted using the following equation: Hemoglobin ed (mg/mL) H emo y 1C1 t' I 11dex —_ x 100 obin t (mg/mL) Where: Hemoglobin Released (mg/mL) = (Optical y x X Coefficient+ nt) x 16 Hemoglobin Present (mg/ml) = Diluted Blood 10 :: 1 mg/mL

[0568] The corrected hemolytic index was calculated by subtracting the hemolytic index of the PBS blank solution from the hemolytic index of the test article and controls.

The test article is compared to the negative control by cting the hemolytic index of the negative control from the hemolytic index of the test article.

Table 6D: Test Parameters: BloodT 6 Used: Nitrile Glove Material, tested at 3 cmZ/mL Pol u-ro lene Pellets tested at 0.2 ; ams/mL Total Hemo lobin Kit: Stanbio, 80 m/dL Incubation Time: Incubation Temerature: 37 :: 2°C Example 100 Minimum Elation Media WEN!) elution test 0fCZ—25 y: The Minimal Essential Media (MEM) n test was designed to determine the cytotoxicity of extractable substances. The test article was added to cell monolayers and incubated. The cell monolayers were examined and scored based on the degree of ar destruction. All test method acceptance criteria were met. The test procedure(s) listed above were followed t deviation.

Table 7A: Results of Test Results Scores mum“ O‘CZ'ZS Pass/Fail Initial Fail 4 4 4 4 Concentration 2.5 m_/mL ————— ————— ————— ————— Table 7B: Controls: Scores Amount . Tested / Extraction Identification Extraction #1 #2 #3 Average Ratio Solvent Amount Negative Control - 0.2 g/mL 4 g / 20 mL Pol IOI'O lene Pellets Media Control “nun—— Positive Control - 4 4 4 4 0.2 g/mL 4 g / 20 mL Latex Natural Rubber Acceptance Criteria: The United States Pharmacopeia & National Formulary (USP <87>) states that the test article meets the requirements, or receives a g score (Pass) if the reactivity grade is not greater than grade 2 or a mild reactivity. The ANSI/AAMI/ISO 10993—5 standard states that the achievement of a numerical grade greater than 2 is considered a cytotoxic , or a failing score (Fail).

Acceptance criteria were based upon the ve and media controls receiving "0" vity grades and positive controls receiving a 3-4 reactivity grades (moderate to ).

The test was considered valid as the control s were within able parameters.

The cell monolayers were examined microscopically. The wells were scored as to the degree of discernable morphological cytotoxicity on a ve scale of 0 to 4: Table 7C: Standards for Grades Conditions of All Reactivity Cultures No cell lysis, None 0 intrac o lasmic _ranules.

Not more than 20% Slight 1 rounding, occasional lysed cells. rounding, no extensive cell 1 sis. roundin; andl sed cells. destruction.

[0574] The results from the three wells were averaged to give a final cytotoxicity score.

Procedure: The test article was added to 1X Minimal Essential Media + 5% bovine serum at an initial concentration of 2.5 mg/ml. This initial concentration was then diluted 1: 2, 1:4, 1:8, and 1:16. Multiple well cell culture plates were seeded with a verified quantity of industry rd L-929 cells (ATCC CCL-l) and incubated until imately 80% confluent. The test articles and control ts were added to the cell monolayers in triplicate. The cells were incubated at 37 :: 1°C with 5 :: 1% C02 for 48 :: 3 hours. e 101 Minimum Elation Media (MEM) elution test 0fCZ-86 Summary: The polyamine compound CZ-86 was added to cell monolayers and incubated according to the procedure described in Example 100.

Table 8A: Results , _ Results mm of (31-86 Pass/Fan 400 m ————— ————_— ————_— ——-_‘--_-_ ——-_-_-_-_ Table 8B: Controls: Amount Tested / Extraction Identification Extraction Average Ratlo Solvent Amount Negative Control - 4 g / 20 mL Polypropylene Pellets Media Control -_ 20 ml Positive Control - 4 4 4 g / 20 mL Latex l Rubber The cell monolayers were examined microscopically. The wells were scored as to the degree of discemable morphological cytotoxicity on a ve scale of 0 to 4: Table 8C: Standards for Grades Cultures No cell lysis, None 0 intracytoplasmic granules.

Less than or equal to 20% Slight 1 rounding, occasional lysed cells. than or eual to 50% rounding, no extensive cell lysis.

Greater than 50% to less Moderate 3 than 70% rounding and l sed cells.

Nearly te Severe 4 destruction of the cell la ers.

Example 102 Minimum Elation Media (MEA/I) elutian test 0fCZ—52 and CZ—100 Summary: The test articles shown below was added to cell monolayers and incubated according to the ure of Example 100.

Table 9A: Results Results Scores Identification Pass/Fail I(a)#2 Avera 1 e a:>—I ’— 4 4; A 4 m(/3 VJ [\J [\J [\J m(/1 (/1 a:(ll (/1 wA-h DJ-h-h ;_a ;_a Hang-l; #-l> N [vb-DA N-b-b-b N-b NA N-b #-l> AAOO 4; A ##-I>OO --_—____ Code for Test Article: 1) Chlorhexidine gluconate 2) Glutaraldehyde 3) CZ-52 4) CZ—lOO ) ycin 6) Benzethonium chloride Table 9B: ls: -S—or-es Amount Tested Extraction Identification -A / Extractlon. verage Ratio Solvent Amount Negative Control - 0.2 g/mL 4 g / 20 mL Pol r0 1 lene Pellets Media Control “nun—— Positive Control - 4 4 4 4 0.2 g/mL 4 g / 20 mL Latex Natural Rubber The cell monolayers were examined microscopically. The wells were scored as to the degree of nable morphological cytotoxicity on a ve scale of 0 to 4: Table 9C: Standards for Grades Conditions of All Cultures Reactivi Grade intrac o lasmic _ranules. rounding, occasional lysed cells. rounding, no extensive cell 1 sis. roundin andl sed cells. destruction.

Example 103 Minimum Elation Media (MEM) elution test 0fCZ-58, CZ-62, CZ—65 and CZ-66

[0580] Summary: The test articles shown below was added to cell monolayers and incubated according to the procedure of Example 100.

Table 10A: Results fi- Results Scores Dil“tion cation Pass/Fail #1 #2 #3 Avera_e t—NAAHb—KN-h-hb—tb—KUJA-hl—tb—KN l—‘N-b-bl—‘l—‘N-b-bl—‘l—‘LQ-b-bl—‘l—‘N j—A h—K Table 10B: Controls: Amount Tested fication / Extraction Average E11323?“ Solvent Amount Negative Control - 0 Wm Pol pro o lene s 0 N/A Positive Control - 4 0.2gmL 4g/20mL Latex Natural Rubber The cell monolayers were examined microscopically. The wells were scored as to the degree of discernable morphological cytotoxicity on a relative scale of 0 to 4: Table 10C: Standards for Grades Conditions of All Cultures Reactivi Grade No cell lysis, intrac o lasmic _ranules.

Not more than 20% rounding, Slight 1 occasional l sed cells.

Not more than 50% rounding, Mild 2 no extensive cell 1 sis.

Not more than 70% rounding Moderate 3 and lysed cells.

Nearly te cell Severe destruction.

Example 104 Minimum Elation Media (MEM) elution test 0fCZ—92, CZ—96 and CZ-99 Summary: The test articles shown below was added to cell monolayers and incubated according to the procedure of Example 100.

Table llA: s Identlfi- s Dilution catlon e /mL Fail 4 4 CZ-92 2 2 0 0 CZ-96 0 0 0 0 0 0 CZ—99 Amount Tested Extraction Identification / Extraction Ratio Solvent Amount ve l - 4 g / 20 mL Polypropylene Pellets Media Control 20 ml Positive Control - 4 g / 20 mL Latex Natural Rubber The cell monolayers were examined microscopically. The wells were scored as to the degree of discernable morphological cytotoxicity on a relative scale of 0 to 4: Table llC: Standards for Grades Conditions of All Cultures No cell lysis, None intrac o lasmic _ranules.

Less than or equal to 20% Slight rounding, occasional lysed cells.

Greater than 20% to less than or equal to 50% rounding, no extensive celll sis.

Greater than 50% to less than Moderate 3 70% rounding and lysed cells.

Nearly complete destruction Severe 4 ofthe cell la ers.

While this invention has been described in n embodiments, the present invention can be further modified within the spirit and scope of this disclosure. This application is therefore intended to cover any variations, uses, or adaptations of the ion using its general principles. Further, this application is intended to cover such departures from the present disclosure as come within known or customary ce in the art to which this invention pertains and which fall within the limits of the appended claims.

Claims (52)

CLAIMS :

1. A erapeutic method for dispersing or killing a biofilm, the method comprising a step of contacting the biofilm with an anti-biofilm composition, thereby dispersing or killing the biofilm; wherein the anti-biofilm composition comprises a biocidal polyamine compound selected from the group consisting of Ra Ra A1 A5 A1 A5 A3 Ra , Ra A3 Ra , and a salt thereof; A1, A2, A3, and A5 are each an independently selected CR5; each Ra is an independently selected group of a V: R1a R1b N R2c R4 Rm R2d R3 ; each R1a and R1b is a member independently selected from hydrogen and alkyl; each R2a, R2b, R2c and R2d is a member independently selected from the group consisting of hydrogen, alkyl, and fluoroalkyl; each R3 is a member independently selected from the group consisting of , -Z1-Y1-R4, -Y2-R4, and -Z1-Y1-Y2-Y3-R4; each Y1, Y2, and Y3 is an independently selected group of Formula IA: Rm R2d each Z1 and Z2 is an independently selected -N(R4)-; each Rm is -CH2-; 16584570_1 (GHMatters) P41348NZ01 each m is an integer independently selected from 1 to 2; each R4 is a member independently selected from the group consisting of hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, cycloalkyl, aryl, arylalkyl, cycloalkylalkyl, and heteroarylalkyl; or, alternatively, for an -N(R4)2 group, one of the two R4 in the group is a member selected from the group consisting of -(CO)OR6a, -(CO)N(R6a)(R6b), and -C(NR6a)N(R6b)(R6c); or, alternatively, for an -N(R4)2 group, the two R4 groups join to form a heterocyclic ring; each R5 is a member independently selected from the group consisting of hydrogen, alkyl, hydroxyl, alkoxy, aminoalkoxy, alkylamino, alkylaminoalkoxy, alkenyl, alkynyl, aryl, aryloxy, arylamino, cycloalkyl, cycloalkoxy, cycloalkylalkoxy, cycloalkylamino, cycloalkylalkylamino, heterocyclyl, heterocycyloxy, heterocycylamino, halo, haloalkyl, fluoroalkyloxy, heteroaryl, heteroaryloxy, arylamino, arylalkyl, arylalkyloxy, arylalkylamino, heteroarylalkyl, heteroarylalkyloxy, arylalkylamino, yalkyl, aminoalkyl, and alkylaminoalkyl; and each R6a, R6b, and R6c is a member independently selected from hydrogen and alkyl; wherein if R4 is -(CO)OR6a, R6a is alkyl; n the ine compound comprises at least six primary or secondary amino groups.

2. The method of claim 1, wherein: each R1a and R1b is hydrogen; and each R2a, R2b, R2c and R2d is hydrogen.

3. The method of claim 1 or 2, wherein: each Ra is an independently selected H(CH2)n]pNHR4; each n is an integer independently selected from 3 or 4; and each p is an integer independently selected from 1 to 3.

4. The method of any one of claims 1 to 3, n the biocidal polyamine nd is a hydrochloride salt.

5. The method of any one of claims 1 to 4, wherein the biocidal polyamine compound is selected from the group consisting of 16584570_1 (GHMatters) P41348NZ01 Ra Ra Ra Ra R5 , Ra , and a salt thereof; wherein: each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is ndently selected from 3 or 4; each p is independently selected from 1 to 3; each R4 is a member independently selected from the group consisting of hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, arylalkyl, cycloalkylalkyl, and heteroarylalkyl; and R5 is a member independently selected from the group consisting of hydrogen, alkyl, hydroxyl, alkoxy, aminoalkoxy, alkylamino, alkylaminoalkoxy, alkenyl, alkynyl, aryl, aryloxy, arylamino, cycloalkyl, cycloalkoxy, cycloalkylalkoxy, cycloalkylamino, cycloalkylalkylamino, heterocyclyl, heterocycyloxy, cycylamino, halo, haloalkyl, fluoroalkyloxy, heteroaryl, heteroaryloxy, heteroarylamino, arylalkyl, arylalkyloxy, arylalkylamino, heteroarylalkyl, heteroarylalkyloxy, heteroarylalkylamino, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; n the polyamine compound comprises at least six primary or secondary amino groups.

6. The method of any one of claims 1 to 5, wherein: each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is 3; and each p is independently ed from 1 to 3.

7. The method of any one of claims 1 to 6, wherein each R4 is a member independently ed from the group consisting of hydrogen, alkyl, fluoroalkyl, alkenyl, alkynyl, arylalkyl, and cycloalkylalkyl.

8. The method of any one of claims 1 to 7, wherein each R4 is a member independently selected from the group ting of hydrogen, alkyl, kyl, and cycloalkylalkyl. 16584570_1 (GHMatters) P41348NZ01

9. The method of any one of claims 1 to 8, wherein each R4 is a member independently ed from the group consisting of hydrogen, isopropyl, isobutyl, hexyl, octyl, and cyclohexylmethyl.

10. The method of any one of claims 5 to 9, wherein each R4 is not hydrogen.

11. The method of claim 10, wherein each R4 is independently selected from the group consisting of butyl, isobutyl, hexyl, and octyl.

12. The method of any one of claims 1 to 11, n each R5 is selected from the group consisting of hydrogen, alkyl, hydroxyl, alkoxy, aminoalkoxy, alkylamino, alkylaminoalkoxy, aryl, y, cycloalkyl, cycloalkoxy, cycloalkylalkoxy, halo, fluoroalkyl, fluoroalkyloxy, heteroaryl, arylalkyl, arylalkyloxy, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl.

13. The method of any one of claims 1 to 12, n each R5 is a member independently ed from the group consisting of hydrogen, hydroxyl, , alkylaminoalkoxy, cycloalkoxy, cycloalkylalkoxy, halo, heteroaryl, arylalkyloxy, and hydroxyalkyl.

14. The method of any one of claims 5 to 11, wherein R5 is hydroxyl, alkoxy, cycloalkoxy, heterocycyloxy, arylalkyloxy, heteroaryloxy, or heteroarylalkyloxy.

15. The method of any one of claims 5 to 12, wherein R5 is selected from the group consisting of hydroxyl, alkoxy, cycloalkoxy, and arylalkyloxy.

16. The method of claim 5, wherein the biocidal ine compound is selected from the group consisting of Ra Ra R5 and a salt thereof; wherein: each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is independently selected from 3 or 4; 16584570_1 (GHMatters) P41348NZ01 each p is independently selected from 1 to 3; each R4 is a member independently selected from the group consisting of hydrogen, alkyl, arylalkyl, and cycloalkylalkyl; and R5 is selected from the group consisting of hydrogen, hydroxyl, alkoxy, alkylaminoalkoxy, cycloalkoxy, cycloalkylalkoxy, halo, heteroaryl, arylalkyloxy, and hydroxyalkyl; n the polyamine compound comprises at least six primary or secondary amino groups.

17. The method of claim 16, wherein each n is 3.

18. The method of claim 16 or 17, wherein each R4 is a member independently selected from the group consisting of hydrogen, isopropyl, isobutyl, hexyl, octyl, and cyclohexylmethyl.

19. The method of any one of claims 16 to 18, wherein each R4 is not

20. The method of claim 19, wherein each R4 is independently selected from the group consisting of butyl, yl, hexyl, and octyl.

21. The method of claim 16, wherein R5 is selected from the group consisting of hydroxyl, alkoxy, cycloalkoxy, and arylalkyloxy.

22. The method of claim 5, wherein the biocidal polyamine compound is ed from the group consisting of Ra Ra Ra and a salt thereof; wherein: each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is 3; each p is independently selected from 1 to 3; and 16584570_1 ters) P41348NZ01 each R4 is a member independently selected from the group consisting of hydrogen, alkyl, arylalkyl, and cycloalkylalkyl; wherein the polyamine compound ses at least six primary or secondary amino groups.

23. The method of claim 22, n each R4 is independently selected from the group consisting of hydrogen, butyl, isobutyl, hexyl, and octyl.

24. The method of claim 5, wherein the biocidal polyamine nd is selected from the group consisting of Ra Ra Ra and a salt thereof; wherein: each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is 3; each p is independently selected from 1 to 3; and each R4 is a member independently selected from the group consisting of alkyl, arylalkyl, and cycloalkylalkyl; n the polyamine compound ses at least six primary or secondary amino groups.

25. The method of claim 24, wherein each R4 is independently selected from the group consisting of butyl, isobutyl, hexyl, and octyl.

26. The method of any one of claims 1 to 4, wherein the biocidal polyamine compound is selected from the group consisting of: 16584570_1 (GHMatters) P41348NZ01 H H H2N N H2N N H BocHN N HN N HN N H H HN N N NH2 N NH2 H H . 6 HCl . 6 HOBz N NHBoc , , H , HN NH2 H HN N H2N N H NH2 NH NH NH HN N N H2N H . N HN N NH2 H 6 HCl , H , , H NH2 H H N N N HN H2N N NH HN H N NH2 H2N HN HN NH H2N HN H2N NH BocHN . N NH NH 9 HCl H NH2 . . mixture of mono / di / tri-Boc 9 HCl , HO 6 HCl , , H H N N H BocHN N NH H2N N NH H H H2N N NH H2N HN H2N HN H2N HN N NH N NH N NH H H H NHBoc . H2N . . 8 HCl x HOBz H2N x decanoic acid , , , 70_1 (GHMatters) P41348NZ01 HN N HN NH NH NH N HN NH2 H O N S N N N N H H H2N H H H NH2 H2N . 8 HCl NH2 . NH2 · 6 HCl 6 HCl , , , N NH N NH H H NH NH O N N O NH H N N NH2 H H H2N H2N NH2 . . 6 HCl H2N 6 HCl , , , HN NH HN NH N N O H H O N HN NH HN HN N H H O N H2N H2N NH2 . . 6 HCl H . 6 HCl 6 HCl , , , N N H H N N NH HN H H NH HN . 6 HCl · 6 HCl NH N NH2 H2N , , 70_1 (GHMatters) P41348NZ01 H H N N H H N N N N H H NH HN NH NH H2N HN NH N 6 HCl · 6 HCl , , N N H H HN NH NH HN HN NH HN NH NH N N N H H H NH HN H2N NH2 · 6 HCl · 6 HCl · 6 HCl , , , N N HN NH HN NH H H NH HN N N O N N H H H H HN HN NH HN NH2 H2N · 6 HCl · 6 HCl , , , HN NH N N N N H H H H O N N N N N N H H H H H H Br NH2 H2N NH HN NH HN NH . . · 7 HCl 6 HCl 6 HCl , , , 16584570_1 ters) P41348NZ01 N NH NH N N H H N NH HN N N H H N N H H · 6 HCl · 6 HCl , , and N N H H NH HN N HN 6 HCl

27. The method of claim 16, wherein the biocidal polyamine nd is selected from the group consisting of: H H H2N N H2N N H H2N N HN N HN N H H HN N N NH2 N NH2 H H . 6 HCl . 6 HOBz N NH2 , , H , N NH2 HN N H H HN NH HN NH NH HN NH2 N S N N N H H . H H NH2 H2N . 8 HCl HO 6 HCl NH2 · 6 HCl , , , 16584570_1 (GHMatters) P41348NZ01 N NH N NH H N NH H NH H NH O N O NH H O NH H2N H NH2 . NH2 . H2N 6 HCl 6 HCl H2N , , , HN NH HN NH N N O H H O N HN HN H N N N H H H NH OH H2N H2N NH2 NH2 . . N 6 HCl 6 HCl 6 HCl H , , , N N H H N N N N NH HN H H H H NH HN NH HN . 6 HCl O O · 6 HCl NH N H2N H2N NH2 H2N 6 HCl , , , N N HN NH H H HN NH NH HN HN NH N N H H NH N N N NH HN H2N NH2 · 6 HCl · 6 HCl · 6 HCl , , , 70_1 (GHMatters) P41348NZ01 N N H H NH HN HN NH HN NH NH N O H N N O N N H H H H NH HN NH2 H2N · 6 HCl · 6 HCl , , , N N H H HN NH NH HN N N O H H O N N N N H H HN HN H H NH2 H2N NH HN NH . · 6 HCl · 7 HCl 6 HCl , , N NH N N NH H H O N N H H H Br N N H H NH HN 6 HCl · 6 HCl , , , N N H H N N H H NH HN NH HN N N N HN H H . · 6 HCl 6 HCl , and .

28. The method of claim 27, wherein the biocidal ine compound is selected from the group consisting of: 16584570_1 (GHMatters) P41348NZ01 N NH2 H HN NH N NH HN NH H H O N S N N H H H2N H . NH2 H2N . NH2 HO 6 HCl · 6 HCl 6 HCl , , , N NH N NH H H NH NH O N N O NH H H N N NH2 H H H2N H2N NH2 . . 6 HCl H2N 6 HCl , , , HN NH HN NH N N O H H O N HN NH HN HN N H H O N H2N H2N NH2 . . 6 HCl H . 6 HCl 6 HCl , , , HN NH HN NH N N H H O NH HN HN NH N N O H H · 6 HCl N N NH HN NH2 H2N H2N NH2 · 6 HCl · 6 HCl , , , 70_1 (GHMatters) P41348NZ01 N N H H NH HN HN NH HN NH NH N O N N O H N N H H H H NH HN NH2 H2N · 6 HCl · 6 HCl , , , N N H H HN NH NH HN N N HN HN H H NH2 H2N · 6 HCl · 7 HCl , , N NH N N NH H H O N N H H H Br N N H H NH HN 6 HCl · 6 HCl , and .

29. The method of claim 19, wherein the biocidal polyamine compound is selected from the group consisting of: 70_1 (GHMatters) P41348NZ01 HN NH N N N N H H H H NH HN NH HN N HN . 6 HCl NH NH N H NH N . N 6 HCl H · 6 HCl , , N N H H HN NH NH HN HN NH NH N O N N H N N H H H H NH HN NH HN · 6 HCl · 6 HCl , , , , N N H H NH HN N N N N O H H H H N N N N H H H H HN HN Br NH HN NH HN . . · 6 HCl 6 HCl 6 HCl , , , 16584570_1 ters) P41348NZ01 N NH NH N N H H N NH HN N N H H N N H H · 6 HCl · 6 HCl , , and N N H H NH HN N HN 6 HCl

30. The method of claim 21, wherein the biocidal polyamine compound is selected from the group ting of: N NH N NH H N NH H NH H NH O N O NH H O NH H2N H NH2 . NH2 . H2N 6 HCl 6 HCl H2N , , , HN NH HN NH N N O H H O N HN HN N H N N H H H NH OH H2N H2N NH2 . N NH2 . . 6 HCl 6 HCl 6 HCl H , , , 16584570_1 (GHMatters) P41348NZ01 HN NH N N N N H H H H NH HN O NH HN N N O O H H · 6 HCl NH HN H2N H2N NH2 H2N 6 HCl , , , N N HN NH H H NH HN N N H H HN HN NH2 H2N · 6 HCl · 6 HCl , and .

31. The method of claim 22 or 23, wherein the biocidal polyamine compound is selected from the group consisting of: 16584570_1 ters) P41348NZ01 H2N N NH HN H NH2 HN N NH2 NH NH NH H2N HN N NH N H2N H H . N HN H NH2 . 6 HCl 9 HCl , , , N H H2N N NH H H2N N NH H2N HN H2N HN N NH H N NH H2N . x HOBz H2N . x decanoic acid , and .

32. The method of claim 24 or 25, wherein the biocidal polyamine compound is selected from the group consisting of: HN N NH NH N HN and a salt thereof.

33. The method of any one of claims 1 to 32, n the biofilm comprises an otic-resistant bacterial species.

34. An anti-biofilm composition comprising a biocidal polyamine compound used in the method of any one of claims 16 to 32. 16584570_1 (GHMatters) P41348NZ01

35. The anti-biofilm composition of claim 34, wherein the biocidal polyamine compound is selected from the group consisting of Ra Ra R5 and a salt thereof; wherein: each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is independently selected from 3 or 4; each p is independently selected from 1 to 3; each R4 is a member independently selected from the group consisting of hydrogen, alkyl, arylalkyl, and cycloalkylalkyl; and R5 is selected from the group consisting of hydrogen, hydroxyl, alkoxy, alkylaminoalkoxy, cycloalkoxy, cycloalkylalkoxy, halo, heteroaryl, kyloxy, and hydroxyalkyl; wherein the polyamine compound comprises at least six primary or secondary amino groups.

36. The anti-biofilm composition of claim 34, wherein the biocidal ine compound is selected from the group ting of Ra Ra Ra and a salt thereof; each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is 3; each p is independently selected from 1 to 3; and each R4 is a member independently selected from the group consisting of en, alkyl, arylalkyl, and cycloalkylalkyl; wherein the polyamine compound comprises at least six primary or ary amino groups. 16584570_1 (GHMatters) P41348NZ01

37. The anti-biofilm composition of claim 34 or 36, wherein the al ine compound is selected from the group consisting of Ra Ra Ra and a salt thereof; wherein: each Ra is an independently selected -CH2[NH(CH2)n]pNHR4; each n is 3; each p is independently selected from 1 to 3; and each R4 is a member independently selected from the group consisting of alkyl, arylalkyl, and cycloalkylalkyl; wherein the polyamine compound comprises at least six primary or secondary amino groups.

38. The iofilm composition of any one of claims 34 to 37, wherein the biocidal polyamine compound is selected from the group consisting of: H H H2N N H2N N H H2N N HN N HN N H H HN N N NH2 N NH2 H H . 6 HCl . 6 HOBz N NH2 , , H , N NH2 HN N H H HN NH HN NH NH HN NH2 N S N N N H H . H H NH2 H2N . 8 HCl HO 6 HCl NH2 · 6 HCl , , , 16584570_1 (GHMatters) P41348NZ01 N NH N NH H N NH H NH H NH O N O NH H O NH H2N H NH2 . NH2 . H2N 6 HCl 6 HCl H2N , , , HN NH HN NH N N O H H O N HN HN H N N N H H H NH OH H2N H2N NH2 NH2 . . N 6 HCl 6 HCl 6 HCl H , , , N N H H N N N N NH HN H H H H NH HN NH HN . 6 HCl O O · 6 HCl NH N H2N H2N NH2 H2N 6 HCl , , , N N HN NH H H HN NH NH HN HN NH N N H H NH N N N NH HN H2N NH2 · 6 HCl · 6 HCl · 6 HCl , , , 16584570_1 ters) P41348NZ01 N N H H NH HN HN NH HN NH NH N O H N N O N N H H H H NH HN NH2 H2N · 6 HCl · 6 HCl , , , N N H H HN NH NH HN N N O H H O N N N N H H HN HN H H NH2 H2N NH HN NH . · 6 HCl · 7 HCl 6 HCl , , N NH N N NH H H O N N H H H Br N N H H NH HN 6 HCl · 6 HCl , , , N N H H N N HN NH2 H H NH HN NH2 NH NH HN N N N HN N H2N H H . H . · 6 HCl 6 HCl 6 HCl , , , 16584570_1 ters) P41348NZ01 H H N N H2N N NH H2N N NH H H HN N NH NH H2N HN H2N HN N NH N NH N HN H H H NH2 . H2N . 9 HCl x HOBz , , , H2N N NH H HN N NH NH H2N HN N NH H N HN H2N . H x decanoic acid , and .

39. A non-therapeutic method for inhibiting formation of a biofilm, the method comprising a step of treating planktonic bacteria with the anti-biofilm composition of any one of claims 34 to 38, thereby inhibiting incorporation of the planktonic bacteria into the biofilm.

40. The anti-biofilm composition of any one of claims 34 to 38, r sing a carrier.

41. The anti-biofilm composition of claim 40, wherein the composition is a tablet, a pill, a , a e, an aerosol spray, a solution, a suspension, a gel, a paste, a cream, a foam, a wash solution, a dressing, a wound gel, or a synthetic tissue.

42. The anti-biofilm composition of claim 40, wherein the composition is coated on or impregnated into a surface of a medical device. 16584570_1 (GHMatters) P41348NZ01

43. The anti-biofilm composition of claim 42, wherein the medical device is a clamp, a s, a scissors, a skin hook, tubing, a needle, a retractor, a scaler, a drill, a chisel, a rasp, a saw, a catheter, an orthopedic device, an artificial heart valve, a prosthetic joint, a voice prosthetic, a stent, a shunt, a pacemaker, a surgical pin, a respirator, a ventilator, and an ope.

44. The anti-biofilm composition of claim 40, wherein the anti-biofilm composition is selected from the group consisting of a sanitizing wipe, a cleanser, a toilet bowl , a shampoo, a bath additive, a liquid soap, a solid soap, a lotion, a cream, a deodorant, a moist cleaning cloth, an oil, and a powder.

45. The iofilm composition of claim 40, wherein the anti-biofilm composition is a paint, a pipe coating, a flush solution, or pipeline flush solution.

46. Use of an anti-biofilm composition in the cture of a medicament for therapeutically dispersing or killing a biofilm on a surface of a subject, wherein the antibiofilm ition comprises: (i) a pharmaceutically acceptable carrier; and (ii) a therapeutically effective amount of a biocidal polyamine compound used in the method of any one of claims 1 to 32; wherein the anti-biofilm composition is ated for topical, oral, transmucosal, vaginal, or rectal administration.

47. The use of claim 46, wherein the biofilm comprises an antibiotic-resistant bacterial species.

48. The use of claim 46 or 47, wherein the anti-biofilm composition is a tablet, a pill, a troche, a capsule, an l spray, a solution, a suspension, a cream, a foam, a liquid, a gel, a paste, or a powder.

49. The use of any one of claims 46 to 48, n the surface is a dermal surface, a mucosal surface, an oral surface, a urinary tract surface, a vaginal tract surface, or a lung e. 16584570_1 (GHMatters) P41348NZ01

50. The use of any one of claims 46 to 49, wherein the e is skin or soft tissue that has been mised by dermatitis, ulcers, a burn injury, or trauma.

51. The use of any one of claims 46 to 50, wherein the anti-biofilm composition is for treating a patient with a biofilm-related disorder.

52. The use of claim 51, wherein the biofilm-related disorder is an infection, pneumonia, cystic fibrosis, otitis media, a urinary tract disorder, a periodontal disease, bronchiectasis, dental caries, or acne. 73919818V.1 16584570_1 (GHMatters) P41348NZ01