NZ761261A - Treatments for resistant acne - Google Patents

Abstract

The present disclosure relates generally to formulations comprising besifloxacin as an active component for the treatment of acne, in particular acne caused by infections with drug-resistant strains of Propionibacterium acnes (Cutibacterium acnes), methods for the preparation of such formulations and their use.

Description

TREATMENTS FGR RESTSTANT ACNE RELATEB CATTONS lll This application claims benefit priority of ot‘lndian Patent Application No. 269/DEL/2tll4, filed January 29, 20M and No. 324:”ii/D‘Eds/ZGlilya filed November l0, 20M, the content of both ations is incorporated herein by reference in their entirety.

FEELB 9F TEE TNVENTEQN lfl lZl The present disclosure relates generally to novel molecules, compositions, and formulations for treatment of bacterial infections in general and more specifically to bacterial infections with antibiotic—tolerant pathogensl BACKGRGUND OF THE ENVENTTON {lllltlitl Acne vulgaris is a skin condition that affects over 85% of all people.

Acne is a term for a medical condition of plugged pores typically occurring on the face, neck, and upper torso Following are four primary factors that are tly known to contribute to the formation of acne vulgaris; (1) increased sebum output resulting in oily, greasy skin; (2) increased bacterial ty, normally due to an overabundance of Propionibacterium acnes bacteria; (3) plugging (hypercornification) of the follicle or pilosebaceous duct; and (4) and inflammation. The plugged pores result in blackheads, whiteheads, pimples or deeper lumps such as cysts or s. Severe cases of acne can result in permanent scarring or disfiguring.

NU] liltilléll Though acne vulgaris is multifactorial, a conunensal skin ia (P. aches) plays a major role in the formation of acne lesion. it is an infection of hacious glands, oil glands in the skin. in most cases sudden breakouts of acne can be correlated with sudden increased production of sebum in the affected individual. During adolescence androgen hormones play a crucial role. it leads to overproduction of sebum by the baceous gland. The situation gets fin‘ther accentuated by irregular shedding of dead skin from lining of hair follicles. As the dead skin cells clump er in the oily nment, they can form plugs which block the pores of the hair follicles. A pore clogged by the shedding skin is referred to as a . {tiling} This creates a very conducive anaerobic condition for P. cones bacteria to grow. Hyperproliferation of P. cones leads to ction of follicular walls and it sends a danger signal to the host immune system. P. acnes may trigger an innate immune reaction both in very early comedogenic) and in late (inflammatory) acne lesions via the activation of Toll like receptors 2 (TLRIZ) on inante immune cells. TLR activation ultimately triggers the expression various l0 cytokines (like lL—o, iL—tt, lL—lZ, lL-l’? etc) and chemokines that stimulate recruitment of other host immune cells [Jeremy et al, 2003; t et al, 20%}.

Acne lesions range in severity from eads, whiteheads and pimples to more serious lesions such as deeper lumps, cysts and nodules. {sass} Although s over-the—counter products are commercially l5 available to counteract acne ion, such as anti—acne agents for l use, including salicylic acid; sulfur; lactic acid; glycolic acid; pyruvic acid; urea; resorcinol; Nsacetylcysteine; retinoic acid; isotretinoin; tretinoin; adapaiene; tazoretene; antibacterials such as clindamycin, tetracyclines, and erythromycin; vitamins such as folic acid and nicotinamide; minerals such as zinc; benzoyl peroxide; octopirox; san; azelaic acid; phenoxyethanol; phenoxypropanol; and flavinoidsa these agents tend to lack in potential to mitigate the acne condition and may have negative side effects when devised in conventional topical formulations A key challenge that has limited the use of topical formulations is the absence of formulations with the desired physicochemical properties and high drug loading, which maintains a concentration significantly higher than the MK: at the site of application by facilitating the right degree of penetration over time but with minimal. systemic exposure. A formulation that addresses these unmet needs can be a significant advance in the treatment of acne: {6997} Furthermore, as articulated in {’l‘aglietti et al, 2068}, when it comes to the delivery of a drug to a specific site, topical formulations that are efficacious are ly among the most challenging products to develop. Once the product is applied on the skin, a complex ction occurs between the formulation, the active compounds, and the skin itself. The penetration of the active compound(s) into the sltin follows Fick’s first law of diffitsion, which describes the transfer rate of solutes as a function of the concentration of the various ingredients, the size of the treatment surface area, and the permeability of the skin. However, the skin’s permeability can be influenced by many factors, such as the drying, moisturizing, or occluding effects of the excipients in the formulation, which, in combination, can modulate the release of the t at the treatment site. in acne, the site of action is inside the pilosebaceous unit and, therefore, an efficacious anti—acne ll) formulation should facilitate the penetration of the active ndfs} into this extremely ilic nment. An effective l formulation therefore needs to provide a stable chemical environment in a suitable dispensing container in order to odate multiple compounds that may have different, if not incompatible, physicochemical characteristics [Tagleitti et al, 2008]. Gnce d, a topical ation must interact with the skin nment, which can influence the rate of the release of the compound(s) in order to achieve adequate shin absorption, and exert additional physical effects on the skin, such as drying, occluding, or moisturizing leitti et al, 2968]. For example, even if an active agent is very potent, and is effective via a systemic route, in the ease of topical administration can behave completely differently, i.e. if the desired concentration is not reached in the pilosebaceous (or skin) unit, it will not serve as an effective anti—acne therapy. rly, a le or drug can behave entirely differently if formulated with different compositions, which we demonstrate later in an example. Similarly, two molecule or active agents may behave entirely differently in the same formulation or composition. Therefore, every new molecule that needs to be formulated for topical skin application poses a novel and independent challenge as it is impossible to predict which composition. and ratio of active and excipients will provide the desired efficacy benefit, {Guild} Furthermore, an emerging condition is the ion of strains of P. cones, which do not respond to theantibiotic agents such as clindarnycin, tetracyclines and erythromycincurrently approved for the treatment of acne.While the earlier dogmawas that antibiotics failure arises due to selection of ‘resistant’ strains, i.e. a mutation resulting in alteration of the target of the antibioticrendering it ineffective, emerging evidence ts that antibiotic failure is more complex than this simple understanding. The assumption was that if resistance develops, he the target of an antibiotic is altered, it is possible to treat the condition by changing to an alternative antibiotic, the target of which is still intact in the bacteria. However, recent knowledge has rendered this assumption as false. For example, Regoes et al, 2064 demonstrates that even in the absence of any resistance, a subset of bacteria can just exhibit tolerance to an l0 antibiotic, i.e. not o lysis. This could arise due to physiological (metabolic) and morphological changes observed in bacteria exposed to antibioticsFor example, in a study published in Science, r et al, 2004} showed that a ent induction of SOS response by ampicillin can protect E. coé’z' against the bactericidal effects of ampicillin. Regoes et al, 2094 ted that nce mechanisms could cross over between some antibiotics, i.e. Antibiotic A might be rendered ineffective due to development of resistance, but it is possible that otic B, which’has an entirely different target/mechanism of action, and is shown to be active in a different or sensitive bacterial strain, may also be rendered ineffective in the resistant strain due to shared tolerance mechanisms. indeed, 2% massive changes in gene expression leading to tion in the syntheses of proteins of metabolic and stress response pathways and cell on during re of E. coil to ampicillin and ofloxacin have been observed, and a number of these tions in the gene expression levels were shared between bacteria exposed to ampicillin and ofloxacin, suggesting a bacteria not respondingto ampicillin may not respond to cin although both agents have different targets, We saw a similar observation in screening a library of antibiotics against different strains of P. genes that are ive or non—responsive to mycin (a lincosamide), As shown in Fig. 1A, the strain of P. common—responsive to clindamycin also showed increased survival capability in the presence of roxithromycin (a macrolide), which targets a different site from clindamycin.

[Keren et al, 2604}, Specialized ter cells and the mechanism of multidrug tolerance in Escherichia coii. l. Bacteriolilgozglflusmfl) ted that random fluctuations in gene expression are responsible for the formation of specialized persister cells. As argued by Regoes et al, 2004, phenotypic tolerance to antibiotic could actually prevent clearance. As a result? while there remains a need in the art L]! for compositions? formulations and methods for treating acnethat is not responding to the currently used agents, ally cliitdainycinw, minocydine—, omycinw, mdjor doxycycline, the ility of tolerance makes itimprobable to predict a drug that may work against 1‘" acnes. l9l Furthermore, it is singly becoming evident that subtle changes in l0 chemical structure of a molecule can dramatically change activity of the molecules against target protein. For example erythromycin (a macrolide) and clindamycinhind to similar SGS ribosomal unit but crystal structure [Schulzen et al, 290i} showed different mode of binding between the agents and amino acid residues present in SOS ribosomal sub—unit. There are known bacterial strains of l5 P. cones that are resistant to clindamycin but can be either non-responsive or susceptible to erythromycinand vice versa. interestingly, romycin, which is a semisynthetic derivative of erythrontycin works well in a bacterial strain that is resistant to both erythromycin and clindamycinW et al, 2003}. Similarly; in another example, the introduction of S-chloro group dramatically enhanced the 2t) potency of moxiflosacin but a similar change in gatifloxacin had no effect againstfi curses, S. pneumonia, and E, coil. Additionally, molecules of the same class can have different affinity for the same protein target but in different bacterial For example it has been found that both besifloxacin and oxacin effectively bind to DNA gyrase than ciprofloxacin in Si ln contrary ciprofloxacin binding towards DNA gyrase is more effective in Ecoli than moxifloxacin or besifloxacin. Similarly, oxacin is found to be best effective le against S pneumonia followed by oxacin and ciprofloxacin.

[Cambau et al, Ztltlgjilt is therefore not le to predict the activity of a moleculeagainst a bacteria or microbe based on its similarity in structure another drug that shows activity against the same microbe or a different microbe, even though they might have similar mechanisms of action indeed, as shown in Figure.

WO 14666 l, we observed that molecules that were verisimilar in structure had completely distinct activity against P. acnes, i.e. where one was inactive the other was very potent against both clindamycinususceptihle and =nonuresistant P. acne: (Fig. l A and l3) in another example, which we discuss later, we ed a non- U1 lincosarnide molecule that was very effective in a P. genes strain ant to clindamycin but not active in a clindamycin—sensitive P. acnes (Fig. lA and lB).

The identification of an effective drug that works against both sensitive as well as clindamycin—, minocycline-, erythromycin—, or doxycycline—nonresponder P. acnes therefore s through serendipity during systematic screening in P. l0 acnes. {0010} Furthermore, while an emerging problem is the development of resistant strains of microhes that are not responding to antimicrobial compounds and compositions well known in the art, there remains a need in the art for a more effective antibiotic that not only works against ant microbes but also reduces the risk of development of resistance by the microbes to this new antibiotic. Thus molecules that are efficacious antibiotics and also ‘prevent’ or reduce the development of resistance can he a major advancement in the treatment of microbial diseases. {00.13.} The inflammatory character of acne has been correlated with the host immune response targeting Propionibaeterium genes, in vitro s demonstrate that P. genes whole cells or cell fractions stimulate cytolrine and matrix metalloproteinase release from immune cells, keratinocytes, and sehocytes [Kim et al, 2002; Liu et al., 2005; Nagy et ah, 2006; Lee et al, 20l0l Though P. acnes are long been present in the follicular area come in , they direct contact with immune cells in dermis only after follicular e, The innate immune system recognizes P. acnes via TLR2 {Kim et al., 2002}, leading to the ion of inflammatory cytolcines, including malt-a ,lL-l2 etc. Follicular rupture happens very late in the disease process. But there are multiple evidences which suggest that the ve immune response also has a significant role in the inflammation ed even in early stages of acne, resulting from the tment of activated T helper l (Thl) lymphocytes to early acne lesions {Mouser et al, 2003}.A ial treatment of acne therefore needs to resolve inflammation, and should be able to target these inflammatory pathways. {$912} An ideal treatment for acne therefore needmolecules that can work at two or more targets. Molecules that work against both antibiotic-sensitive as well, as clindamycinw, minocycline—, erythromycin— and cline-tolerant or ponsivestrains of P. acnesand can additionally inhibit the P. genes-activated inflammatory mediator/s, or molecules that target two or more cellulm targets in these microbes while additionally ng an inhibitory effect on the P. acne;- activated inflammatory mediator/s, and is formulated in an optimal formulation it? that enables the desired concentration of the active agent on the skin or pilosehaceous region following topical application can emerge as a powerful strategy for the treatment of acne. {9&3} A series of novel DART (Dual Action Rational Therapeutics) molecules were designed and synthesized for treatment of bacterial infections caused by both susceptible and resistant gram positive and gram negative bacteria and specially for curing acne and different skin and skin structure ions and additionally prevent the development of resistance. DART molecules can mount its activity h two distinct mechanisms of action in a micro‘oe (such as a bacteria), and create less chance in mutation development at both target sites in the bacteria. Additionally, it can also act at the host level by ting the immune response, such as altering the levels of inflammatory ines. {iltlldl The design of DART comprises of two active domains. The two active domains can he selected from different families, for examplefi—lactam, B—lactam derivatives, 2- and 4—ouinolones, quinolones having halogenated atom specially fluorine atom attached at (3-6 or {3—7 on of the central ring system, tluoroquinolone with halogenated atom specially chlorine atom attached at {3—8 3G position of the l ring system, ycline, oxazolidinone, hydroxypyridones, derivatives of hydroxypyridones, plenromutilin, azoles, nitroimidazoles, monoxycarboiic acid eiass, c acid, suifonamide, suifonamide derivatives, retinoids, different fatty acids (saturated, unsaturated), propyiene giycoi and glycerol derivatives of different fatty acids and a strategic combination from each of the famiiies. The design was made strategicaiiy by arranging the two active Vi domains in the right steric arrangement for both the active domains to maintain their function against bacteria or fungus. Overaii these moieeuies possess faster ial hitting with reduction in ation and activity against ant nathogens. These moiecules aiso show a lower risk of pment of resistance. {dfiifii in some embodiments, the BART moiecnie has at least two ai it} domains. Each of said chemicai domains binds to a distinct or different active site in a target celi. in a preferred embodiment, a third chemical domain may be present. in a r preferred embodiment, said two chemicai domains may be bound together through a said third domain. in some embodiments, the BART moiecuie has at ieast two distinct or different anti—bacterial mechanisms of action. in some embodiments, the BART moiecuie has at least two distinct or different cne mechanisms of action. Without iimitations, the BARTs can act on the same target or on different targets, for exampie, the bacteria and the host. in some embodiments, the BART acts on at ieast two different s. in some embodiments, at ieast one of the targets is different than that affected by conventionai antibiotics.

{Mild} In some embodiments, the BART moiecuie has a B—iactam ring and a quinoione nucieus, or a quinoione s and a nitroxheterocycie, or a B—iactam ring and a nitroheterocycie. {9&7} In some embodiments, the BART has at ieast two distinct anti— bacterial mechanisms of action, for e inhibits DNA gyrase or topoisomerase W and transpeptidase-mediated cross~iinhing of peptidogiycans; inhibits isoprenyi pyrophosphate and eptidase-mediated crosswiinking of peptidoglycans; inhibits isoprenyi pyrophosphate and DNA gyrase of topoisomerase EV; inhibits foiate synthesis and DNA gyrase of topoisomerase 1V; 3G inhibits foiate synthesis and transpeptidasednediated crossdinhing of peptidoglycans; inhibits DNA gyrase or topoisomerase 1V and the 303 ribosomai it in ia; inhibits DNA gyrase or topoisomerase iv and the SOS sub~ unit in bacteria; inhibits transpeptidase—mediated linking of peptidoglycans and the 303 or the SOS ribosomal sub-unit in bacteria; inhibits folate synthesis and the 308 or the SOS sub—unit in bacteria; or ts isoprenyl pyrophosphate and U1 the 308 or the SOS sub—unit in bacteria, or causes DNA modification, such as inducing DNA nickswhile inhibiting the induction of negative supercoils in DNAgor altering the fluidity of the cell membrane while exerting an activity on the DNA; or altering the levels of metal ions in a cell while inducing DNA changesin some embodiments, the first mechanism of action is an anti—bacterial action and the second mechanism of action is anti-inflannnatory or immnnomodulatory. {961% in some embodiments, the DART molecule has at least two ct treating acne mechanisms of action and modulates at least two different targets in some embodiments, the first mechanism is an antibacterial action and the second mechanism of action is inhibition of keratinocyte proliferation and differentiation. in some embodiments, the DART molecule has two distinct acne treating isms of action and whereinthe first mechanism is an antibacterial action and the second mechanism of action is anti~intlammatory, in some embodiments, the DART molecule is effective against forms of Proptonbacterium acnes that respond poorly to clindamycin~, or doxycycline—, or erythromycin~, or minocycline—containing anti—acne products. in some embodiments, they areeffective t one or more of clindamycin—, minocycline-, erythromycin—, and/or doxycycline~ tolerant or resistant s of Propionbacteriam genes. in. some ments, it prevents the development of resistance in P. genes. {36195 in some embodiments, the DART molecule has at least two distinct anti—bacterial mechanisms of action and modulates at least two different targets against a pathogen. Non limiting examples of suchpathogens are : Bartonella henselae, Sorrelia burgdorjf‘erij Campylobaeter jejuni, Campyiobacterjfbtus, Chlamydia trachomatis, dia pneumoniae, Chylamydia psittaci, Simkania 3t) negevensis, Escherichia coli (ag, li7 and K88), Ehrlichia chafiensis, Closi‘ridium botulinum, Ciostridium pcrfiingens, Closa‘ria’iam temni, Enterococcas WO 14666 faccalis, Haemophilt’us zae, hiiius ducrcyi, Coccidtoides immitis, Bordctciia pertussis, Coxicila burnetii, Urcapiasma urcat’yticum, .: ycoplasma t’tum, Trichomatis vaginalis, Hcfz'cobacter pylori, Helicobacter hepaticus, Legionella pacumophila, Mycabacterium tubercuiosis, Mycobactcrium bavis, (J‘ Mycabacterium afiicatzum, Mycaaactcrium leprae, fiftfi/‘cabactcrtum asiaticam, Mycobacterium avium, fvfycabacteriun'z celatum. Mycobactcrium ceianae, Mycobactcrium fortuitum, Mycottacterium genavcnsc, Mycobactcrium haemophiium, Mycobactcrium intracellularc, Mucobacterium katzsasii, micabactcrium matmocusc, Mycobactcrium marinum, Mycobactcrium scrofitlaccum, Mycobacterium simtae, Mycobactcrium szulgai, Mycobactcrium ut’ccrans, cterium xenapi, Corynebocterium diptheriac, Rhodococcus cam”, Rickcttsia aescklimanuii, Rickettsia afi‘icae, Rickcttsia conorii, Arcanobacterium yticum, Bacillus anthracis, us cercus, ia monocytogerzes, Yerst‘nia pestz‘s, t’crstnia cntcracotitica, Shigcila dysenteriac, Ncisscria meningitides, Ncisscria ganorrhccac, Streptococcus bovis, Streptococcus hemoiyticus. Streptococcus mutam‘, Streptococcus pyagenes, Streptococcus pacumaniae, Staphylococcus aurcus, Staphylococcus cpia’ermidis, Staphylococcus pneumoniac, Staphyt’acoccus saprophyticus, Vibrio cholerac, Vibrio cmolyticus, Salmonelia typhz’, Salmoncila pht, Salmonet’la tia’is. Treponema pallidum. Candida, Cryyjtcooccus, Cryptosporia’ium, Gtara’z’a Zambiz'a, Microsparidia, a’ium vivax, Pneumacystt‘s carinii, Toxoplasma gondii, Trichophyton mentagrophytes, Enteracytozoon bieneusi, pora cayctaneusts, Encephalitozocn heliem, Encephalitozoon cuaiculi, among other bacteria, archaea. protozoa? and fungi. 33929} In some embodiments, the first and second domains independently have cteriai activity against a Staphylococcus spocies. Examples. of Staphylococcus spades include, but are not iimited to, S. aarcus group tag, S. aureus. S. simiac), S. auricularis group (cg, S. auricularis), S. carnosus group (cg. S. us, S. condtmcnti, S. massit’iensis, S. pisciflermentans, S. simulans), S. cpidermictis group (cg, S. capitis, S. caprae, S. midis, S. saccharoiyttcus}, S. haemalyticus group (cg, S. csci, S. haemot’yticus, S. hominis), S. hymns—intermediate group (cg, S. chromogenes, S. felts, S. deénhini, S. hyicus, S. intermedius, S. lutrae, S. microti, S. mascara, S. pseudiniermedius, S. rostri, S. schiei'ferz), S. lugdunensis group (eg, S. Zuga’unensis), S. soprophyticus group (cg, S. arietiae, S. cobalt, S. m, S. galfinarum, S. kloosiz’, S. Zeei, S. nsis, S. saprophyticus, S. us. S. s), S. sciuri group (3.3:, S. fleureaii‘, S. icnius, S. sciuri, S. siepanovicii, S. newlines), S. simulates group (cg, S. tes), and S. warneri group (eg, S. pasteuri, S. warneri). {6&2le Without limitations, the DARTS can be in the form of particles, powders, suspensions, dispersions, emulsions, liposonies, micelles, globules, solutions, vesicles, aggregates, , gels, and the like. {fidZZl The disclosure also provides formulations comprising DARTs as the active pharmaceutical ingredient (APB. otics {$923} The disclosure also provides formulations comprising antibiotic agents, which are not DARTs, as the APlln some embodiments, the antibiotic agent is aS—chloro fluoroquinolone. Exemplary S—chloro lluoroquinolones include, but are not limited to, hesifloxacin, clinafloxacin and sitatloxacin. In some embodiments, the ation comprises besilloxacin as the APl. 2t} {lltlZdl in various embodiments, the APl can be micronized, suspended, or solubilized. in some embodiments, the APl can be in the form of particles, s, suspensions, dispersions, emulsions, mes, micelles, globules, solutions, vesicles, aggregates, and the like in some embodiments, the API can in the form of a drug carrier. {tltlZS{ in some embodiments, the APl can be coated. in some other embodiments, the APi can be uncoated. {titlZél Without limitations, the formulation can be in a form ed from the group consisting of lotions, creams, gels, emulgel, oils, serums, powders, sprays, ointments, solutions, suspensions, dispersions, pastes, foams, peels, films, masks, 3t) patches, sticks, rollers, cleansing liquid washes, cleansing solid bars, pastes, foams, powders, shaving creams, impregnated fabric ), and the like. in some embodiments, the formulation is in a form ed from the group consisting of gel, cream, spary, face wash, soap bar, body wash, lotion, suspended drug loaded gel, suspended drug loaded cream, and any combinations thereof. {$327} in some ments, the APl or the formulation can be used to treat acne not ding to antibiotics. Specifically, it exerts greater cy against forms of Propionbacterium cones that respond poorly to clindamycin—, or doxycycline—, or omycin—, or minocycline-containing anti-acne products. {$328} in some embodiments, the Add or the formulation can be used to treat acne by exerting an anti—inflammatory effect. {@329} in some embodiments, the APl or the formulation can be used to treat acne by killing s of Propionbacterium gene that are sensitive toone or more of clindamycin—, minocycline-, erythromycin—, and/or doxycyclineand additionally exerting a greater efficacy by inhibiting P. acnesnnediated inflammatory pathways (ie. dual mechanisms of action). ltll The disclosure also provides formulations comprising a combination of two or more antibiotic agents. For example, an Suchloro quinolone in combination with another anti-acne agent. in some embodiments, the formulation 2t) comprises two or more different 8~chloro fluoroquinolones. ln some embodiments, the formulation comprises besifloxacin and a retinoid, such as {6931} in some embodiments, the formulation comprises an anti—acne agent and an anti—inflammatory agent. For example, the formulation can comprise an ti— chloro fluoroquinolone and an antiwinflammatory agent. {@932} in some embodiments, the two or more antibiotic agents can be a DART molecule or two or more different DAR’l‘ molecules. ln some embodiments, one of the two or more antibiotic agent is a BART and the other is not a DAR’I‘ le. {limit} As described herein, the disclosure provides formulations comprising DART and/or non-DART antibitotic agent as the APl. As such, exemplary APl’s for the formulations include DARTS, antiwhacterial, anti-fungal and anti—acne agents, in some embodiments, the API can he in the form of a drug carrier, tie, the APl can be nanotized, coated, made into vesicles, liposorne, emulsions, and the like for the formulation. Without limitations the formulation or the composition can he formulated for administration by any appropriate route known in the art including, but not limited to, topical ( ing huccal and sublingual) and oral or parenteral routes, including intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, and nasal administration. {@934} The DARTS and formulations disclosed herein can he used for treating bacterial infections due to Gram—positive or Gram-negative bacteria Additionally, the DARTS are effective against resistant forms of ens. rmore, the DAR’l‘s are effective in preventing the development of resistant forms of pathogens. Thus, the DAR’ls and formulations disclosed herein can be used for treating antibiotic tolerant or resistant bacterial infections. Exemplary l5 bacterial infections include, but are not limited to, infection by Barroheiia ae, Borreiia harga’orferi, Campylahacter jejani, Campylahacteigfeius, Chlamydia matis, Chlamydia pneumoni‘ae, Chyiamydia psiitaci, Simkaiiia hegevehsis, Escherichia caii (eg, 0157zll7 and K88), Ehrlichia chafeensis, Ciastridiam hum, Ciastria’ium perjiririgehs, ria’ium terani, Enteroeoeeus faecaiis, Haemophiiius influenzae, Haemophiiius dacreyi, Coceiriioia’es immiiis, telia sis, {l'oxieiia burnetii, Ureaplasma ureaiyiieam, ,tfiicoplasma genitaiium, Triehomaris vagihaiis, Helicoha’eier pylori, Helicobacier hepaiieus, Legioneiia pheamophiia, Mycahacteriam tuberculosis, eierium basis, Myeobaeterium africaham, Mycobacierium ieprae, Mycobaeterium asiaiieam, tidycebaeterium avium, iiiycobacterium ceiaiam, .hfizcohacteriam eeionae, @Icobaeierium fortuiium, Mycahacierium rzse, Myeohaeierium haemophiiam, Myeobaeierium iniraceh’ar’are, ,hhicohacterium kahsasii, Mrcabacterium maimoense, Mycobaezerium marinam, eierium laeeum, Myeebaererium Simiae, llaiycobacterium szuigai, cierium 3G ulcerahs, Mycahaeieriam xenopi, bacierium dipiheriae, Rhadoeaecus eqai, Rickeiisia aesehlimahrzii, Ricketisia afi‘icae, Rickettsia canorii, Areahobacreriam haemolyricum, us anthracis, Bacillus cereus, Lysteria monocytogenes, Yersinia posits, Yersinio enterocolitico, Slzigella oysenterioe, Neisserio meningirides, Neisscrio gonorrhoeoe, Streptococcus bovis, Streptococcus hemolvticns, Streptococcus melons, Streptococcus pyogenes, Streptococcus S pneumoniae, Staphylococcus oureus, Staphylococcus epidermidis, Staphylococcus nioe, Staphylococcus soproplzyticns, Vibrio cholerae, Vibrio emolyricus, Salmonella n/phi, Salmonella paratyphi, Salmonella cnteritio’is, Treponemo pallidum, Candida, ertptcooccus, Cryptosporldium, Giordia lamblia, Microsporldio, Plasmoa’ium vita/wt, Pneumocystis carinii, it) Toxoplasmo gondli, Triclzoplzylon menragropltytes, Enterocytozoon blenensi, Cyclosporo coyetonensls, Encephalitozoon hellem, alitozoon coniculi, among other bacteria? nrehaeaa protozoa, and fungi. in some embodiments, ion is with a Staphylococcus species. {@935} in some ments, the DARTS and formulations disclosed herein can. he used for treating acne. in some embodiments, the DARTS and formulations disclosed herein are effective against forms of Propionbacterium acnes that respond poorly to clindarnycin—, or doxycycline-, or erythromycin—, or minoeyeline—eontaining anti—acne products. in some embodiments, the DARTs and ations sed herein are effective against one or more of 2G elindamyein-, minocyelinen, erythrom.yein-, and/or doxyeyeline— tolerant or resistant s ol‘Propionbacrcrium acnes. For treating infections, the BART or the formulation disclosed herein can be stered once or daily to the t as a single dose or a plurality of doses.

BRIEF BESQ‘REPTiGN GF THE DRAWENGS geese} Figs 1A and iii shows dose response curves of different antibiotics against both MTCCl951 and CCARM 95)“) strains of P. ncnes. While MTCClQSl is killed by clindamycin, the drug has no effect on CCARMQGlO. ent antibiotics behave differently and unpredictably on the different strains belong to a different family from Clindsmyein. 11111371 Figs. 11: and 11) are line graphs showing concentration efficacy curve of DART nds 911, 91. 141, 113, 115 and 116 in both clindarnycin- susceptible (MTCC 1951) (Fig. 1C) and clindamycinunonresponsive (CCARM 9010) (Fig 1.1)) P. times s. The les have different and ictable activity against MTCCl951 and CCARM9010 strains of P. acnes. Compound 91 showed highly efficacious bacteriai killing profile for both bacterial strains.

Compound 911 shows activity in P. acnes that do not respond to clindamycin but is ineffective in the P. genes strain that responds to clindamycin. 111 Figs. 2A and EB show concentration—dependent inhibition of DNA gyrase activity (super—coiling) by nd 91. Fig. 2A - Agarose gel electrophoresis showing effect of compound 91 on coiling of E. 6031' plasmid DNA by DNA Gyrase. Fig. 23 entage of DNA super—coiling by DNA gyrase in presence increasing concentrations of compound 91. {111139} Figs. 3A is a bar graph showing percentage of DNA super-coiling by DNA gyrase in presence of compounds $11, $113 94, 113, 115 and 116 with relaxed E. coli plasmid DNA. nd 91and compound 116 seemed to have the best gyrase inhibiting activity among all the comparators. Though this observation is mostly correlated with MlC data agianst P. cranes, yet there is some s» specific advantages is observed with compound 91. 111114111 Fig. 313 is a bar graph showing percentage of DNA super-coiling by DNA gyrase in presence of compound 91 and nadiiloxacin with relaxed E, coir” plasmid DNA. Compound 91 showsgreaterefficacy than Nadifloxacin. 11111411} Figs. 41A. and 41% are bar graphs showing the effect of compound 91 on P. acnesuinduced cytokine 1L — 6 (Fig. 41A), 11 .. 8 (Fig. ~13) release in THP-vl cells. Compound 91 exerts an anti~inilamrnatory activity against P. acnesdnduced cytokine production. Statistical analysis was performed using Student’s t~test (* p = 0.05; M“ p = 0.005). 11111421 Figs. 5A and SB are bar graphs showing the effect of compound 91 on P, acnes~induced cytokine 1L — 1n (Fig. 5A), 1L — 16 (Fig. 53) release in THP—1 cells. {0043} Fig. 6. is a bar graph showing the minimum inhibition value for some exemplary topical gel formulations against P. aenes. {0044} Fig. 7 is a line graph showing the close response curve of Zone of inhibition (ZOE) of some exemplary gel formulations t P. acnes.

U1 E0045} Fig. 8 is a line graph showing the time kill kinetics of some exemplary gel ations t P. oer-res. {0046} Fig. 9 Graph shows the efficacy of a l formulation of besitloxacin in P. cones in an in viva skin infection model. Besitioxacin gel formulation has the ability to clear almost 1.5 log CFU (~95%) of inoculum of clintlamycin resistant P. acnes within first 24 hours. {0047} Fig. iii is a line graph showing time kill cs of some exemplary besifioxacin formulations against P. genes MTCC 1951, showing that the composition of the formulation can change the efficacy of an antibiotic. {0048} Fig. iiis a line graph showing time kill kinetics of some exemplary l5 besitioxacin formulations and besitloxacin APis against S. oureus. {0049} Fig. i2 is a line graph showing time hill kinetics of besifloxacin against P. apnea (CCARM 9010). {0050} Fig. i3 Graph shows that topical formulations with different ent compositions for the same antibiotic can result in different profiles in the skin and in systemic circulation of SD rats. Fully suspended 1% Besitioxacin gel (VLN— Fig/BSF/GL/(log), fully soluble l% oxacin gel (VLN-FZl/BSF/GL/OOlA) and fully suspended 1% Besifloxacin gel (VLN—FZtl/BSF/CR/Gw) were used for comparison purpose. To be efficacious, a formulation should not only be physicochemically compatible with the antibiotic but also enable a susmined concentration of the antibiotic at a concentration greater than MIC level.- 30051} Figs MA anti itiii are bar graphs showing the concentrationw ent inhibitory effect of Besifloxacin on P. genes—induced cytohines iL—é (Fig. 14A) but not 1&8 (Fig. MB) release in l cells. Statistical analysis was performed using Student’s f-tcst (* p = 0.005; ** p == 0.0005). {0052} Figs. ESA anti ESE are bar graph showing the combination of Besifioxacin and Adapalene ses the efficacy of inhibiting P. genes—induced WO 14666 cytohines lL-o (Fig. ESA) but has no effect on lL—rS (Fig. 153) release in THP~l cells. Statistical analysis was med using Student’s Host (’3‘ p 2 0.005; ** p = acct).

Ei) EESCREPTEGN 0F THE iNVEN’i‘EGN {@953} Acne vulgaris is a skin condition that affects over 85% of all people.

Acne is a term for a l condition of plugged pores typically occurring on the face, neck, and upper torso. Following are four primary factors that are currently known to contribute to the formation of acne vulgaris; (l) increased sebum output l0 resulting in oily, greasy skin; (2) increased bacterial activity, normally due to an undance of Propionibacrcrium cranes bacteria; (3) plugging (hypercornification) of the follicle or pilosebaceous duct; and (4) and ation. The plugged pores result in blackheads, whiteheads, pimples or deeper lumps such as cysts or nodules, Severe cases of acne can result in l5 permanent scarring or disfiguring. {@8354} As articulated in htt ://theseienceofacne.eom’antibiotic-susce tibilit - oil-propionibacterium—acnes/ results from studies over last four decades clearly demonstrate that over time P. aortas bacteria has become increasingly resistant to certain classes of antibioticsPartieularly important are observations that a significant tage of the bacteria isolated from acne patients are now resistant to the most common antibiotics used in acne treatment: Clindamycin, Erythromyein, Tetracycline, Doxycycline and Minocycline. Additionally, such resistant or antibiotic—tolerant strains can cause relapse of acne, and also cause other diease states. There is a need for antibiotics that can kill P. acnes while zing the probability of development of mutant or tolerant strains that can survive the antibiotic exposure, and those that can work against strains that are not responding to the current drugs. Additionally, if these novel les can target additional steps in acne formation, such as inflammation, then the clinical outcome in acne can be greater than existing therapies, {9955} Skin is a major organ of the body, and performs many essential functions besides acting as a barrier, such as maintaining homeostasis. Besides acne, there are many other skin diseases that are caused by bacterial colonization of the skin. The most common bacteria for mild to moderate skin infection are Staphylococcus and Streptococcus e.g., Acute Bacterial Skin. and Skin Structure infection (ABSSSl), Such ia can infect the skin of both pediatric and adult patients; mainly develop during hospitalization or living in a g home, while gardening, or while swimming. Some people are at particular risk of developing skin infections, for example, people suffering with diabetes, human it) immunodeficiency virus (HIV) or AlDS or other immune ers, or hepatitis, and who is undergoing chemotherapy or treatment with other drugs that suppress the immune system. {9956} Common skin bacterial infections e cellulitis, erysipelas, impetigo, folliculitis, and les and carbuncles. Cellulitis is a painful, l5 erythematous infection of the dermis and subcutaneous tissue characterized by warmth, edema, and advancing s and is usually caused by Streptococcus or Staphylococcus species. Erysipelas is a superficial form of cellulitis with sharply demarcated borders and is caused almost exclusively by Streptococcus. impetigo is also caused by Streptococcus or Staphylococcus and can lead to lifting of the 2% stratum corneum resulting in the commonly seen bullous effect, Folliculitis is an mation of the hair follicles, and it is most commonly caused by Staphylococcus, if the infection of the follicle is deeper and involves more les, it moves into the furuncle and cle stages and usually requires incision and drainage. Two different kinds of skin diseases occurred due to the toxins produced by the ia include, Staphylococcal Scaled Skin me (SSSS) which usually affects children less than 5 years old, adults with kidney failure and the other one is Toxic Shock SyndromeThere is more chance of colonization of S. career is found with patients suffering from eczema and atopic itis, a type of inflammatory, relapsing, non-contagious, itchy skin disorder.

L...) O 'l‘hus Staphylococcus ourcus infection plays an important role in atopic dermatitis (AD) or atopic eczema (Ali). Unfortunately, some strains of Staphylococcus have become resistant to methicillin and other similar otics which are known as MRS/5t. Recently it has been found that more than one-half of all cases of skin bacterial infections caused by MRSA species. The infections associated with MESA species cannot be cured with traditional penicillin—related drugs. instead, MESA must be treated with alternate antibiotics. ’ll However as articulated in littp:/’/thescienceofacnecom/antibiotic— susceptibility—of—propionibacterium-acnes/ “Not all antibiotics are created equal”.

The same is true for bacteria. Some types of antibiotics are highly effective against n types of bacteria, while essentially worthless against others. ll) Moreover? antibioitic susceptibility and resistance is a dynamic process that is constantly changing. Over time, certain. types of bacteria may gain or lose resistance to particular antibiotics. The primary problem with standard laboratory—based antibiotic resistance testing is that the tibility of a ia to an antibiotic is often ent when it is growing on a petri dish versus when it l5 is growing on your body, This is because bacteria are not static organisms, they adapt to their environment. A P. acnes bacteria g in a follicle and g on sebum has a different lic profile than one growing on a petri dish and feeding on a ial nutrition supplement. Furthermore, bacteria modulate expression of surface proteins, cell wall structures and genesdepending on their environment, and these changes can have a profound effect on their susceptibility to a particular antibiotic. As a result, in the ease of topical antibiotics for treatment of skin bacterial conditions, apriori knowledge does not exist, i.e. there is no mechanism to predict that an antibiotic will be effective against P. acnesacnes or any other skin bacterial condition until it has been tested on the bacterial strain.

For e, as shown in http://thescienceofacnecom/antibiotic~susceptibility~of~ propionibacterium~acnes/, P. cones was reported to be highly resistant to a nitroirnidazole (metronidazole) or a ycline (lymecycline) but partially responsive to doxycycline (another tetracycline), and showing no resistance to ciprofloxacin but resistant to another fluoroquinolone, Levofloxacinlt is therefore impossible to predict which antibiotic will work based on a priori activity in other bacterial strains. There is a need for a systematic development of novel antibiotics that show activity against acne. [3958} in this regard DART molecules can act as an ideal drug candidate to acne caused by P. cranes, and additionally for the treatment of other skin and skin ”J! structure infections caused by other bacteria such as MESA. DARTs were ed to contain two distinct chemical domains, selected from different families as mentioned earlier, for example a 5~lactam ring and a quinolone s, or a quinolone nuclues and nitrowheterocycle, or a galactam ring and a nitroheterocycle, which confers two distinct mechanisms of action. This creates l0 less chance in mutation development at both target sites of bacteria resulting in less resistance development against these antibiotics. Some of the molecules cm exert additional anti=inilammatory mechanisms to reduce host infiamniatory se, r enhancing the antiwacne efficacy, {b35533 'l‘he embodiments of the various aspects disclosed herein are based on. l5 the les ed by the inventors, which can act on at least two different or distinct targets. Generally, the molecule includes at least two ent or distinct chemical domains, Each of said al domains binds to a ct or different active site in a target cell. The said chemical domains can be bound together through a third domain. As used herein, the term “chemical domain” means a part of a molecule that is involved in a desired property. For example, a al domain can be part of the molecule involved in binding of the molecule with a target or involved in modulating an activity of the target. t} in some embodiments, the first and second chemical domains independently have anti-bacterial or bactericidal activity. in some embodiments, the first and second domain can ndently comprise an antibacterial agent.

As used herein, the term “antibacterial agent” or “antibiotic agent” is defined as a compound having either a bactericidal or bacteriostatic effect upon bacteria contacted by the compound. As used herein, the term “bactericidal” is defined to mean having a destructive killing action upon bacteria, As used herein, the term “bacteriostatic” is defined to mean having an inhibiting action upon the growth of bacteria. Examples of antibacterial agents include, but are not limited to, macrclidcs 0r kctolidcs such as crydhi‘omycin, azithmrnycin, clarithrcmycin, dirithmmycin, trclcandcmycin, splrarnycina iclithrcmycin, carbcmycin a3 ‘icsamycin, kitasamycin, midccamycin acetate, cniycin, sclithminycin, spiramycin,irclcand0mycin, ccihrcniycin, solithrcinycin, spirarnycin, ansamycin, clcandcmycin, carbomycin and tylcsin; bcta~laciams including llin, ccphalcspcrin and caibapcncms such as carbapcncms imipcnem and mcrcpcncm; monolactarns such as penicillin g, penicillin v, meihicillin, cxacillin, clcxacillim diclcxacillin, nafcillin, ampicillin, azlccillin, amcxicillin, icillim ticarcillin, mczlccilllna pipcracillin, azlocillinfi icmocilling fluclcxacillln, ccpalothin, ii} ccphapirin, ccphraclinc, ccphaloridinc, ccfazclin, ccfamandclc, ccfurcxinic, cxin, zil, ccfaclcn lnracarbcfl tin, azolc, ccfctaximca ccfiizcximc, ccfiriaxcnc, ccfcpcrazcnc, ccfiazidimc, ccfixinic, ccfpcdcximc, ccfiibutcn, ccfdinir, ccfpircmc, ccfcpimc, ccfadroxil, cefalci’hin, ccfalcxin, ccfurcximc, ccfditorcn, ccfiazidirnc, <.:c‘fiizcxiinca ccfiamlinc fcsamil, ccftarclinc, iprclc, aztrccnam, crtapcncifine dciipcncm and ciiastatin; penicillin combinations such as aincxicillin/clavulanaic, ampiclllin/sulhactam, piperacillin’tazcbactam and iicarcillin/clavulanatc; quinclcncs such as nalidixic acid, cxclinic acid, ncrflcxacin, pcficxacin, in, cin? cxacin, ciprofloxacin, tcmaficxacin, lcmcflcxacin, flcrcxacin, grcpaflcxacin, xacin, imvalfi'lcxacina clinaflcxacin, gatiflcxacin, nicxiflcxacin, sitaficxacin, gancficxacin, gcrnillcxacin, pazuflcxacin, besiflcxacin, uliflcxacin, pruliflcxacinfi cincxacin, piromidic acid? pipcmidic acid, rcscxacing mllcxacin, balcflcxacin, tcsuflcxacin, dclaflcxacini ncmcncxacin; antibacterial sulfcnamides and antibacterial sulphanilamidcs, including parawarninchcnzcic acid, sulfadiazinc, silver sulfadiazinc, sulfisoxazclc, sulfamcthcxazolc, sulfadimcthcxlnc, sulfadcxinc, sulfamcthizolc and sulfathalidinc, dc, sulfacciamidc, sulfiscmidinc, snlfanilimidc, sulfasalazinc and sulfcnamidochryscidinc; amincglyccsidcs such as streptomycin, nccmycin, kanamycin, paromycina geniainicin, tchrainycin, amikacin, nctilrnicin, spcc’iincmycin, siscniicini 3C9 lin, icina dihydrcsncptcmycim ctin, ainycin, arbekacin, hckanamycin, dihckacin, hygmmycin b, vcrdamicin and, astromicin; iciracyclincs tetracyciine, etracyciine, demeciocyciine, minocyciine, oxytetracyciinei inethacyciine, doxycyciine, cicmccyciine, lymecyciine, meciocyciine, penimcpicyciine, and roiitetracyciine; rifan‘iycins such as rifampicin (aiso caiied rifarnpin), rifapentine, rifabutin, hezoxazin, crifamycin and rifaxirnin; U5 amides such as iinconiycin and ciindamycin; iipopeptide like daptcmycin; giycopeptides such as vanccrnycin, teiavancin and teicopianin; streptograrnins such as ristin and istin; ansamycins such as geidananiycin, herbimycin, rifaxirnin; oxazciidinones such as iinezoiid, eperezoiida posizciid, radezciid, ranhezoiid, sutezoiid and tedizciid; pieuromutiiins such as retapamuiin, i0 iin, vainen‘rulin; steroid antibacterials such as insidic acid; amnheniccis such as chiorantphenicoi, azidatnfenicoi, thiamphenicoi, fiorfenicci; nitrofurans such as furazoiidone, nitrcfurantoin; streptogramins such as pristinamycin, ristin/daifopristin virginiarnycin; other antihacteriais such as arsphenarnine, fosfomycin, mupirccin, piatensitnycin, tigccyciine, trirnethoprini, poiymyxin, hacitracin, colistin, eciyrnycin, inetronidazcie, cctrirnoxazoie and phosphonornycin; and anti—niyeohacterial drugs such as clofazirnine, dapsone, capreornycin, cycicserine, ethanthutoi, ethionarnide, iscniazid, pyrazinatnide, rifarnpicin, rifa‘outin, ntine, streptomycin. in some emhediments, the antihactcriai agent can he sciected from the group consisting of azithron‘iycin, roxithrentycin, ceftaroiine, cefotaxirne, cefcxitin, axone, cephaiothin, minocyciine, oxacin, rnoxifloxacin, hesifioxacin, uiifloxaein, pruiifioxacin, retapamulin, metronidazoie, ornidazoie and any combinations fi in some emhodients, antibacterial agent can he hyaluronic acid or a derivative thereof. {tittéi} in some embodiments of the BART rnoiecuie, the first and second dcntains independentiy have anti~acne activity. in same ernhctiirnents, the first and second chemicai s are independently an anti-acne agent. As used herein, the term “anti—acne agent” refers to any chemical that is ive in the treatment of acne and/or the symptoms associated ith. Anti~acne agents are weii known in the art such as US. Pat. App. Pub. No. 2006/ 0008538 and US.

Pat, No. 5,607,980, content of both of which is incorporated herein by reference. es of usefui anti—acne agents include, but are not d to kcratoiytics, such as salicylic acid, derivatives of salicylic acid, and resorcinol; retinoids, such as retinoic acid, tretinoin, adapalene, tazarotene; sulfur-containing D- and Le amino acids and their derivatives and salts; lipoic acid; otics and antimicrobials, such as benzoyl peroxide, triclosan, chlorhexidine gluconate, (.11 octopirox, tetracycline, 2,4,4'~trichloro—2'-hydroxy diphen‘yl ether, 3,4,4’— trichlorobanilide, nicotinamide, tea tree oil, xib, azelaic acid and its derivatives, phenoxyethanol, phenoxypropanol, isopropanol, ethyl acetate, clindamycin, erythromycin, and ycline; sebostats, such as flavonoids; and bile salts, such as scymnol sulfate and its derivatives, deoxycholate, and e; l0 and combinations thereof. 'l‘hese agents are well known and commonly used in the field of personal care. {$962} in some embodiments, the anti—acne agent can be an antimicrobial peptide having activity against P. cones. Antimicrobial peptides are ubiquitous in nature and play an important role in the innate immune system of many species l5 [Zasloff at at, 2002; and Epand et al., l999]. The antimicrobial peptide can be a naturally occurring peptide or an analog thereof, or it can be a tic peptide.

As used herein an “analog” refers to a naturally-occurring antimicrobial. peptide that has been chemically modified to improve its effectiveness and/or reduce its toxic side effects. The antimicrobial peptide can be a peptide known to be effective against Gram positive bacteria. iting examples include lantibiotics, such as nisin, subtilin, min and galliderrnin; defensins; attacins, such as sarcotoxin; ins, such as cecropin A, bactericidin, and lepidopteran; magainins; melittins; histatins; brevinins; and combinations thereof. Additionally, antimicrobial peptides having activity against P. genes have been reported, for e, in US. Pat. App. Pub. No. 2905/0282755; No. 2005/02455452; and No.2005/6209l57, and US. Pat. No. 279, content of all of which is incorporated herein by reference. Suitable examples of antimicrobial peptides having reported activity against P. genes include, but are not limited to, novispirins haug, supra), and those described in US. Pat. App. Pub. 3t) No.2007/0265431, content of which is incorporated herein by reference. in some embodiments, the antimicrobial peptide can be cathilicidine and its derivatives.

E3963} in some embodiments, the anti—bacterial agent can e fatty acid (FFA) or fatty acid tives or fatty acid esters of propylene glycol (PG) or glycerol(G) derivatives and any ations thereof. For example, lauric acid, stearic acid, myristic acid, oleic acid, linoleic acid, myristoleic acid, palmitooleic U1 acid, linoleic aicd, linolenlc acid, sapienic acid, different saturated FAs (PUFA), propylene glycol monolaurate, glycerol mono and/or di e, propylene glycol monoloeate, glycerol mono and/or di oleate and other derivatives known in art. Fatty acids are wellknown antimicrobial agent a et al., 1972] and their activity varies with chain length, degree of unsaturation and lit number of fatty acid ester present in propylene glycol or glycerol backbones, Theprime target of PAS and derivatives is bacterial cellmembrane, which is non: ic in nature. Bisrnpticn of bacterial membrane causes disruption incellularelectron transport activity or oxidative phosphorylation or inhibition to a particular enzyme activity or diminishingcellular energy production or impairment of nutrient uptake or auto-oxidation of degradation ts or generation of toxic peroxidation ct lysis of bacterial cells. Their broad spectrum of non-«specific activity makes them as a promising antimicrobial candidate for treatment and prevention of antimicrobial infections easued by number of ositive and gram=negative bacteria thatgeneratesvarious skin and skin ure infections. in some embodiments, theFA and derivativesalone or in combination with any antibiotics or covalent conjugates with any otics (cg, DARTS) are effective against antibiotic pronePropionbacterium acnesas well as P. genes that respond poorly to clindamycin—, or doxycycline-, or omycirn, or minocycline~ containing anti-acne products. ln some embodiments, they are effective against one or more of clindarnycin—, minocycline—, erythrornycin—, and/or doxycyclinew tolerant or resistant strains of Propionbacterium genes. in some embodiments, it prevents the development of resistant forms of pathogens lililtidl in some embodiments, the first and second anti-acne agents in the DART or ations disclosed herein are selected independently from the group 39 consisting of acetretin, adapalene, alitretinoin, azelaic acid, Azithrornycin, benzoyl peroxide, lEiesifloxacina bexarotene, Cefotaxime, Cetoxitina Ceftaroline, iprcle, Ceftriaxone, Cephalothin, clindamycin, erythremycin, etretinate, Garencxacin, glycelic acid, isotretinoin, lactic acid, ciine, Mexit‘ioxacin, N—acetylcystein, Nadifloxacin, octepirex, phencxyethanel, phennxypropanol, Pruiifloxacin, pyruvic acid, Radezolid (RX—W41), resercinol, Retaparnulin, (Jr retinoic acid, rcmycin, salicylic acid, Sitaflcxacin, sodium sulfacetamide, nlactone, sulfacetamide, sulfur, tazarctene, tretincin, triclosan, xacin, nietronidazole, cmidaznle, urea, and any combinations thereof, {8665} in some embodiments, the first and second chemical domains are independently an antifungal agent. As used herein, the term “antifnngal agent” is intended to mean a substance capable 0f inhibiting er preventing the growth, ity and/or reproduction of a fungal cell. Preterahle ingai agents are these capable cf preventing or treating a fungal infection in an animal or plant. A preferable antifungal agent is a bread spectrum antifnngal agent. However, an ngal agent can also be specific to one or more particular s of fungus. {8066} Examples of antifiingal agents include, but are not limited t0, azales (e.g., Fluconaznle, lsavuccnazele, ltraconazcle, Ketoconazole, Miccnazcle, Clcrtrimaznle, Vericcnazcle, Pcsacenazole, Ravucnriazcle, Ciclopircx, etc), pclyenes (e.g., natamycin, lucenscmycin, nystatin, amphotericin B, etc.), echinocandins tag, Cancidas), pradimicins (e,g., beancmicins, niidtemycins, 2t) serdarins, allylamines, etc), ’l‘riclesan, Pircctcne and its elamine salt, fenprcpimorph, terhinai‘ine, and derivatives and analogs f. Additional antifungal agents include these described, fer example, in lnt, Pat. Pub, No.

W020Gl/‘066551, No. W02Q62/090354, Ne. W92QOG/043390, N0.

WOZOlG/G32652, No. W02003/GG839l, Ne. 04/018485, N0. lW02065/006869, No. WOZQQS/Q8627l, No. W02002/067889; in US. Pat. App.

Pub. No. 2OQ8/Ol94661, No. 2008/9287449, Ne, NOS/M39943, Ne. 2010/0063285, Ne, EGGS/0032994, No. 047135, Ne. 2068/6l82885; and in US. Pat. Ne. 6,812,238; No. 4,588,525; No. 6,235,728; No. 6,265,584; No. 4,942,l62; and N0, 6,362,l72, t of all of which is incorporated herein by reference. {@967} in some embodiments, the antifungal agent is a pyrithione salt.

Examples of useful pyrithione salts include, but are not limited to, zinc pyrithione, sodium pyrithione, potassium. pyrithione, lithium pyrithione, ammonium pyrithione, copper pyrithione, calcium pyrithione, magnesium pyrithione, strontium pyrithione, silver pyrithione, gold pyrithione, manganese pyrithione, and ations thereof. Non—metal pyrithione salts such as the ethanolamine salt, chitosan salt, and the disulfide salt of pyrithione (which is commercially available as GMADINE M’DS or OMDS), can also be used, The pyrithione salt can be used in any ulate form, including, but not limited to, crystalline form such as platelets, rods, needles, blocks, round and amorphous, regularly or irregularly shaped particles. {6368} in some embodiments, the pyrithione salt is zinc pyrithione. Zinc. ione is best known for its use in treating dandruff and seborrhoeic dermatitis, it also has antibacterial properties and is effective against many pathogens from the Streptococcus and Staphylococcus genera. its other medical applications include treatments of psoriasis, eczema, ringworm, fungus, athlete’s foot, dry skin, atopic dermatitis, tinea, and vitiligo. {@369} in some embodiments, the ngal agent is an antifisngal e.

Antifungal peptides are well known in the art (see for example, De Lucca et al., 2000}. The antifungal peptide can be a naturally ing peptide or an analog thereof, or it can be a synthetic e. As used herein, the term “analog” refers to a naturally occurring antifungal peptide that has been chemically modified to improve its effectiveness and/or reduce its toxic/side effects, Exemplary antifungal peptides can include, but are not limited to, syringoniycins, syringostatins, otoxins, mycins, echinoeandins, pneumocadins, aculeacins, mulundocadins, cecropins, alpha-defensins, efensins, novispirins, and combinations thereof. Other antifungal peptides include those described, for example, in US. Pat. No. 6,255,279 and US. Pat. App. Pub. No. 239799; No, 2005/ 151; No. EGGS/0282755, and No. EGGS/0245452, content all of which is incorporated herein by reference. {$979} in some embodiments, the first al domain is an anti-bacterial agent and the second chemical domain is anti—acne agent or an antihmgal agent, {tltl‘l’l} Without limitations, the first and the second chemical s in the DART can be hound to each other covalently, One of skill in the art is well aware U: of different functional groups in al domains that can be used for covalently g a first chemical domain with a second al domain. For example, the first chemical domain can comprise a functional group selected from the group consisting of an amino group, a N-substituted amino group, a carboxyl group, a carbonyl group, an acid anhydride group, an aldehyde group, a hydroxyl group, an ll) epoxy group, a thiol, a disulfide group, an alkenyl group, a ine group, a hydrazide group, a semicarhazide group, a thiosemicarbazide group, an amide group, an aryl group, an ester group, an ether group, a glycidyl group, a halo group, a hydride group, an isocyanate group, a urea group, a urethane group, and any combinations thereof for binding with the second chemical . in some embodiments, the second chemical domain comprises a fianctional group ed from the group consisting of an amino group, a N—substituted amino group, a carboxyl group, a carbonyl group, an acid anhydride group, an aldehyde group, a hydroxyl group, an epoxy group, a thiol, a disulfide group, an allcenyl group, a hydrazine group, a hydrazide group, a semicarbazide group, a thiosemicarbazide group, an amide group, an aryl group, an ester group, an ether group, a glycidyl group, a halo group, a hydride group, an isocyanate group, a urea group, a urethane group, and any combinations thereof for binding with the first chemical domain, in some embodiments, the first and second chemical domains are bound to each other via same functional group. in some embodiments, the first and second chemical domains are bound to each other via different functional groups. {llll'72} in some embodiments, the third domain can enhance or increase an activity of at least one of the chemical domains~ For example, the ty of at least one of the chemical domains is sed or ed relative to when the third domain is absent. in some embodiments, the third domain can increase or enhance antibacterial activity of at least one of the chemical domains in the BART. In some embodiments, the third domain can increase or enhance anti- acne activity of at least one of the chemical domains in the BART. in some embodiments, the third domain. can increase or enhance anti—inflammatory activity of at least one of the chemical domains in the BART. ltltt’73l in some embodiments, the third domain itself has biological ty.

For example, the third domain can he an active agent. in some embodiments, the third domain can have anti—bacterial or anti-fungal activity. in some ments, the third domain. can have anti-inflammatory activitye ltlll74l The third domain of the DARTS can he a direct bond or an atom such as oxygen or sulfun a unit such as NR1, (3(0), C(Q)O, C(O)NH, SS, SO, $02, ltl SQgNH or a chain of atoms, such as tuted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alhynyl, arylalkyl, arylalltenyl, arylallezynyl, heteroarylalkyl, heteroarylalkenyl, arylalkynyl, heterocyclylalltyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroarylt hetetocyclyl, cycloalkyl, cycloalltenyl, alkylarylalkyl, alltylarylalltenyla alkylat’ylalltynyl, ylat'ylalkyla alkenylarylallcenyl, larylalkynyl, alltynylarylalkyh alkynylarylalltenyl, alkynylarylalltynyl, alkylheteroarylalltyl, alltylheteroarylalkenyl, alkylheteroarylalltynyl, alltenylheteroarylalkyl, alkenylheteroarylalkenyl, alkenylheteroarylalkynyl, lheteroarylalliyl, alltynylheteroarylalhenyl, all<ynylheteroarylalkynyl, alkylheterocyclylalliyl, alltylheterocyclylalkenyl3 alkylhereroeyclylalltynyl, alkenylheterocyclylalltylfi lheterocyclylalkenyl, alltenylheterocyclylalltynyl, alltynylheterocyclylalltyl, alkynylheterocyclylalltenyl, alleynylheterocyclylalkynyl, alkylaryl, laryl, alkynylaryl, heteroaryl, alkenylheteroaryl, alkynylhereroaryh where one or more methylenes can be interrupted or terminated by Q, S, 3(0), S023 N(R§)23 l‘x) LA (3(0),, C(O)O, cleavable linking group, substituted or tituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted hetetocyclic; where R1 is hydrogen? acyl, aliphatic or substituted aliphatic. {@375} in some embodiments, the first and second chemical domains are covalently bound to each other via a third domain comprising at least one cleavable group, A cleavable group is one which is sufficiently stable under a first set of conditions and can be cleaved to release the two baits the cleavable group is holding together. in a preferred embodiment, the cleavable group is cleaved at least it) times or more, ably at least liltl times faster under a first reference condition (which can, e.g., be selected to mimic or represent intracellular conditions) than under a second reference condition (which can, e.g., be selected to mimic or represent conditions found in the blood or serum). l’til Cleavable groups are susceptible to cleavage agents, e.g., pH, redox ial or the ce of ative molecules. Generally, cleavage agents are more prevalent or found at higher levels or activities at the desired site of action of the molecule comprising the cleavable group. Examples of such l0 ative agents include: redox agents which are selected for ular substrates or which have no substrate specificity, including, e.g., oxidative or reductive enzymes or reductive agents such as rnereaptans, present in cells, that can degrade a redox cleavahle linking group by reduction; esterases; arnidases; mes or agents that can create an acidic environment, cg, those that result l5 in a pH of five or lower; enzymes that can hydrolyze or degrade an acid cleavable linking group by acting as a general acid, peptidases (which can be substrate specific) and proteases, and phosphatases.

KNEW} Exemplary cleavable groups include, but are not limited to, redox cleavable groups (cg, -S—S- and ~C(R)g—S-S—, wherein R is H or Ct'Cfi alkyl and at least one R is , alkyl such as CH3 or CHZCH3); phosphateebased cleavable linking groups (e.g., ~t)—P(t))(0R}—O~, —O-P(S)(OR}~O-, —0-P(S)(SR)—O—, —S-— P(O)(QR)—=G—, —OIP(O){OR)—Su, -S~P(O)(0l§)—S—, ~Q—P(S)(O'Rk)-S—, —S—P(S)(OR)— 0—, —G—P(O)(R)—G~, —O-P(S)(R)—Ou, )(R)—O—, uSwP(S)(R)-=Ct—, -S-P(Q)(R)=S~ , ~O~P(S)( R)~S—, . ~0—P(O)(0H)-O-, —O~P(S)(Ol—l)—Gu, —O—P(S)(SH)~O~, is- Pttil)(0l-l)—Ct~, ~O—P(O)(OH)—S-, -S-P(O)(OH)—Sw, =O=P(S)(OH)—S-, =S—P(S)(Oll)— O=, ~G-P(G)(H)~O—, —O—P(S)(H)~G~, -S—P(0)(li)~0—, ~S—P(S)(H)—O~, —S-P(O)(H)— 3-, and -{)~P(S)(l-l)~S—, n. R is optionally substituted linear or branched C;— Cm alkyl); acid cleavable groups (eg, hydrazones, esters, and esters of amino acids, =C=NN- and -OC(0)-); ester-based ble groups (cg, -C(Q)Ov); peptide—based eleavable groups, (cg, groups that are cleaved by enzymes such as peptidases and proteases, e.g., m Ni—icnnt‘crowrtcnafic(o)-, where aA and RB are the R groups of the two nt amino acids). A peptide based cleavable group comprises two or more amino acids, in some embodiments, the peptide— based cieavable group comprises the amino acid sequence that is the substrate for a peptidase or a protease found in ./ secreted by P. genes. {$373} in some embodiments, the hle group is an acid labile group.

Generally, an acid cleavable group is cleavahle in an acidic environment with a pH of about 6.5 or lower (e.g., about 6.5, 6.0, 5,5, 5.0, 4.5, 4.6, 3.5, 3.6, or lower), or by agents such as enzymes that can act as a general acid. {£3679} in some embodiments, the first and second chemical domains are ll) covalently bound together by a third domain selected from the group consisting of roxyundecenic acid; i,lG-decanediol; 1,3—propanediol; l,5—pentanedil; iti— hydroxydecenic acid; succinic; lactic acid; 3—hydroxypropionic acid; and any combinations thereof. {tidbit} in some embodiments, the third domain can be , e.g., a cleavable or non—cleavable linker, {tidbit The first chemical domain can be bound to the second al domain or the domain connecting the first and second chemical s via a direct bond or an atom such as oxygen or snliitr, a unit such as NH, {3(0), C(030, C(0)NH, SS, SO, 80;, or SOlei, {($82} Similarly, the second chemical domain can be bound to the first chemical domain or the domain connecting the first and second chemical domains via a direct bond or an atom such as oxygen or sulfur, a unit such as Nil, C(O), C((DG, C(Q)Nl-i, SS, SO, $02, or SOzNii. {tili83} 'l‘he DARTS can be synthesized using methods known in the art.

Exemplary methods for synthesizing the DAR'l‘s are bed in the Examples section herein. See Examples 2 to it).

{W84} in some embodiments, the DART can be selected from those shown in Tables EA & EB. écwww thx/Hfl m 9m ”Rafi . £34m“ “£4713.Eu 0.: \ .1.

D3/\\(I\Jt\: . a N ......\Dv m . _ alm‘)‘\l‘\\I/\\I(\!I\Q.

“A; z 4m: .cr a :2QCKZ«r @Emymk as WO 14666 WO 14666 WO 14666 WO 14666 WO 14666 WO 14666 mwm<m $3?ny WO 14666 .Euw Eva fig ufiggia exfinsfiz 393% .23” weggaw 325 uEEfi: NM,“ T 233% 3gamma mmfi. fig. ,Eua mfggig utgfi bagging “:3 a u§3fita§ a“$9an {6685} The present inveniton also provities fonnulations comprising DART as the AH. Various features of the formulations are described in more detail vide infra. otics (it‘ifrm—D/lRTil {9986} The present invention also envisages compounds which are not DARTS. Accordingly, the invention also provides formulations comprising an antibiotic agent which is not a DART; 129., a formulation comprising a non—DART antibiotic agent as the API. For example, the present invention describes the use of S-chloro fluoroquinoiones for the treatment of acne ions, especially those caused by resistant forms of P. acnes. it is a sub class of fluoroquinolones where C8 position is substituted with ne. Thus, in some embodiments, the disclosure provides a ation comprising Suehloro fluoroqninolone as the API. Exemplary S—chloro fluoroquinolones includeS but are not limited to, besifloxac-in, sitailoxaein, and elinailoxaein. in some embodiments, the formulation ses oxaein R2 = H, Cyclopmpyl Examples — H2“ H2” For Clinafioxacin Sitafloxaein .............................................................................................................. {(3987} Without wishing to be bound by a theory, micronization of an antibiotic agent, such as besifloxacin, can have an impact on its bioactivity. For example, micronieation can ce antibiotic agent’s bioactivity or its retention U: at a desired site. Further, ization can also affect the antibiotic agent’s stability and amounts in a formulation. Moreover, micronization can also allow optimizing properties of formulations comprising micronized besitloxacin, ingly, without limitations, the APl in the formulation, (eg, the antibiotic agent) can be in the form of particles, s, suspensions, dispersions, ll) emulsions, liposomes, micelles, globules, solutions, es, aggregates, and the like. {@983} in some embodiments, the APl, for example, but not limited to, besifloxacin or DART, can be micronized, tie, formed as a particle.

Generally, the micronized APl has a size in the range from about (32 um to about l5 pm. in some embodiments, the micronized APl has a size in the range from about 1 pm to about l8 unit. In some embodiments, the micronized API has a size in the range from about l.5 um to about 9 um. in some embodiments, the ized APl has a size in the range from about 2 pm to about 8 pm. lfifiggl in some embodiments, the APl is in the form of a particle and comprises a surface modifier on the e thereof. Generally a surface modifier is a molecule that can change the e of the particle (such as by coating) in question and help in adhering the whole particle, hence, to the specific surface(s).

Generally, the surface modification does not involve chemical bonding alterations or creation of any chemical bond. The e modifier just physically associates with the particle. ltlll9tll The surface modifier can be selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, olipids, lipoproteins, cationic molecules, and any combinations thereof.

The surface modifier can form a coating layer on the le surface. Without limitations, the particle can be partially or fully coated with the surface modifier.

WO 14666 ll Some nonnlimiting exemplary formulations sing a ized antibiotic agent, cg, besifloxacin, are described in Examples lS—ZQ and shown in Table 13. {8992} in some embodiments, the formulation can be a spray formulation.

Exemplary non—limiting spray formulations are described in e 23 and fable 1?. ln some embodiments, the formulation can be in the form of a face wash. Exemplary non—limiting face wash formulations are described in Example 24, and Table 2%. ln some embodiments, the formulation can be in the form of a soap bar. Exemplary non—limiting soap bar formulations are described in Example ll} 25 and Table Zl. in some embodiments, the formulation can be in the form of a body wash. Exemplary non—limiting body wash formulations are described in Example 26, and Table 22. in some embodiments, the ation can be in the form of a lotion. Exemplary non-limiting lotion formulations are described in Example 27, and ”l"able 23. [fill93l tants are known to solubilize hydrophobic substances by reducing the interfacial tension. Accordingly, in some embodiments, the antibiotic agent can be solubilized with a surfactant before forming the formulation. lnwaddition to surfactants, co~solvents or co—surfactants can also help in solubilization of the poorly water-soluble compounds by increasing the wetting property or reducing the interfacial tension of the hydrophobic molecule. Some exemplary surfactants and (Lo-surfactants can include, but are not limited to, sodium lauryl sulfate, tween 80, tween 20, span 2t}, and any combinations thereof.

Exemplary co—solvents for lizing the antibiotic agent, such as besiiloxacin, can include propylene glycol. monocaprylate and diethylene glycol monoethyl ether. Additional surfactants, rfactants and cor-solvents that are amenable for solubilizing the antibiotic agent are described elsewhere in the disclosure. t wishing to be bound by a theory, solubilizing the antibiotic agent, cg, besifloxacin, can provide formulations that are within FDA prescription guidelines and limits of inactive excipients or ingredients. Nonnlimiting exemplary formulations sing a solubilized Krill, e.g., antibiotic agent such as hesifloxacin, are described in es 28, El and 33, and shown in Tables 24, 27 and Bil-32. {9994} Preparation of drug—loaded (suspended form) gel via conventional methods usually leads to exposure of the drug to a wide range of pH conditions, which can lead to, in some instances, solubilization of the drug, and then ipitation. This solubilization~reprecipitation phenomenon in most cases leads to change in original particle size, impurity profile or crystal pattern, or . in order to circumvent this issue, the inventors have used an inventive approach to prepare different suspended drug-loaded formulations. 'l‘hus, in some it) embodiments, the formulation is in form of a suspended gel with negligible or minimal. drug solubilization~reprecipitation in the suspended drug formulations, the APl particles are disperesed in a carrier media, such as, but not limited to, glycerol, and processed to the d formulation. ary suspended gel formulations are described in Example 24, 26, 23, 29, 30, 31 and 32 and shown in Tables 25, 2‘7, 29, 33, 34, 37 and 33, Exemplary ded drug loaded cream tormulations are described in es 30 and 32 and Tables 26 and 23. {3&95} in addition to the various components, the formulation can also comprise one or more ity modifiers. in some ments, the viscosity modifier is a polymer. Exemplary polymeric viscosity modifiers include, but are not limited to, carhopol, hydroxypropyl cellulose, hydroxypropyl methyl ose, hydroxyethyl cellulose, and sodium hyaluronate. in some other embodiments, the viscosity modifier is a non-polymeric viscosity modifier or gelling agent, Additional exemplary viscosity modifiers are described ere in the disclosure. Exemplary ations comprising various viscosity modifiers are described in Example 33 and Tahles 29-32, roses} According to published literature there may be some kind of physical. and/or al interaction between carhopol and fluoroquinolones. For which, there may he a need to prepare formulations without carbopol or carhopoldike— polymers to avoid any incompatibility issues during the product shelf life.

Accordingly, in some embodiments, the formulation is essentially free ofd viscosity modifiers. Exemplary formulations that are essentially free of a viscosity modifier are described in Example 32 and Table 28. ltitl97l in some embodiments, the APl can be coated with a molecule selected from the group ting of lipids, oils, polymers, peptides, proteins, carbohydrates, ipids, phospholipids, lipoproteins, cationic les, and any combinations thereof. t limitations, the AH can be partially or fully coated with the coating molecule. ary formulations comprising coated or non—coated Al’l are described in Example 46 and Tables 52 and 53. {3693} it is noted that various formulations features discussed in more detail it) vide infra are applicable to the ation comprising antibiotic agent, e.g., besifloxacin, bed . {6999} in some embodiments, the formulation comprises two or more otic agents, For example, the formulation can comprise two or more different anti—acne agents. in some embodiments, the formulation comprises an 8—chioro fluoroduinolones alone or in combination with another anti—acne agent.

Exemplary 8-chloro fluoroduinolones include, but are not limited to, besitloxacin, sitafloxacin, and clinafloxacin. in some embodiments, the formulation comprises besitloxacin. in some embodiments, the formulation comprises besitloxacin and adapalene, ltlllliltil Without limitations, the two or more antibiotic agents can be in the same form or different forms. For example, the first and second antimbiotic agents can be independently micronizcd, suspended, or solubilized for the API.

Ix) U} Accordingly, in some embodiments, both the first anti-biotic agent and the second anti—biotic agent are micronized, in some embodiments, the first anti-biotic agent is micronized and the second anti—biotic agent is solubilized. in some embodiments, the first anti—biotic agent is micronized and the second anti—biotic agent is suspended in the formulation, In some embodiments, the first iotic agent is solubilized and the second anti-biotic agent is micronized. in some embodiments, both the first anti—biotic agent the second anti-biotic agent are solubilized in some embodiments, the first anti—biotic agent is soiubiiized and the second anti—biotic agent is suspended, in some ments, the first anti-biotic agent is suspended and the second antiwbiotic agent is micronized. in some embodiments, the first antibiotic agent is suspended and the second anti—biotic Ll] agent is solubilized in some embodiments, boht the first anti—biotic agent and the second antibiotic agent are suspended, iiiiiiiiii in some embodiments, the formulation comprises besifloxacin and adapaienea n besifioxacin is soiubilized and adapaiene is suspended. in some ether embodiments, the ation comprises besifioxacin and adapaiene, it) wherein both the besifioxacin and adapaiene are soiubilized. Exempiary ation comprising both besifloxacin and adapalene are described in es i8, 23-27 and 3i and Tahies 13:23 and 27. {961532} it is noted that various formulations features discussed in more detail vide infra are applicable to the formuiation comprising two or more antibiotic agents described herein.

Features aggiicable to DART non—DART and combination APIS 596193} Furthermore? as articulated in http://thescienceofacne.com/antibiotic~ tibiiity-of~propionibacterium-acnesO"i‘he second major iirnitation of treatment of acne is that antibiotics are not evenly dispersed throughout the different tissues in the body: Many antibiotics do not effectiveiy accumulate in the foilicie and/or ous , and therefore do not effectiveiy reach the bacteria responsible for acne. Even if a bacteria is highiy susceptible to a particular antibiotic in lab-based testing, if that antibiotic does not make it to the site of infection at a sufficient concentration? it is not going to be an effective treatment. As a resuit there can be major differences in the effectiveness of orai otics and topical antibiotics used in acne treatments.’ This extends to all bacterial diseases of the skin. There is a need to deveiop unique optimal topical formuiations and is described later. ltltlltlct} Skin, e.g., micro—cracks, sweat or secretion pores, and hair follicles can act as reservoirs for drug carriers of particular sizes. Efficacy of active agents, e.g., antifungal and antibacterial formulations can be enhanced using infimdibttlar delivery. A drug carrier can enhance the delivery of active agent on sebum filled hair follicles and also exhibit necity of such drug carriers to lipophilic microbial cell wall /’ cell membrane. This allows ion of the drag carrier on the shin, followed by slow and continuous release of the DART or antibacterial agent from the drug carrier, Exemplary drug carriers include, but are not limited to microparticles, nanoparticles, vesicles, mes, emulsions, globules, and ll) solutions. tlS} in addition to the APT (cg, DART and/or other antimbacterial agent), the drug carrier can further se one or more additional components. For example, the drug carrier can further comprise a compound ed from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, ic molecules, and any combinations thereof. The Al’l, e.g., DART or other antiwbacterial agent, can be present in the core of the drug carrier and the additional component can form a coating layer over the core. Without limitation,the coating can be a functional or nonfunctional g° By functional coating is meant a g that imparts one or more desirable ties to the drug carrier,snch as enhanced targeting or retention at site of action, increase in the activity or" the APT, or having a destired activity itself. {66166} in some embodiments, DART and/or other anti-bacterial agent can be formed as a particle. in addition to the APT (eg, DART andfor other anti— bacterial agent), the particle can further comprise a nd ed from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations thereof. The APl, e,g,, DART or other anti-bacterial agent, can be present in the core of the particle and the additional component can form a coating layer over the core, {@6137} in some embodiments, the particle comprises a surface modifier on the surface thereof. Generally a surface modifier is a molecule that can change the surface of the particle (such as by coating) in on and help in adhering the whole particle, hence, to the specific surfacet's). Generally, the surface U) cation does not involve chemical bonding tions or creation of any chemical bond. The surface modifier just physically associates with the particle. {illilfltll The surface modifier can be selected from the group consisting of , oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations thereof.

The surface modifier can form a coating layer on the particle surface. Without limitations, the particle can be partially or fully coated with the surface modifier. tl9l in some embodiments, the drug carriers and ations disclosed herein can r comprise an active agent, i.e., an active agent in addition to the BART and/or anti—bacterial agent. As used herein, the term e agent” means a compound or composition that has a particular d activity. For e, an active agent can he a eutic compound. Without limitations the active agent can he selected from the group consisting of small organic or nic molecules, saccharines, oligosaccharides, polysaccharides, peptides; proteins, peptide analogs and derivatives, peptidomimetics, nucleic acids, nucleic acid analogs and 2t) derivatives, antibodies, antigen binding fragments of antibodies, lipids, extracts made from biological materials, naturally occurring or synthetic compositions, and any combinations thereof. liltiiilll ln some embodiments, the active agent can be selected from the group consisting of antifungal agents, antibacterial agents, antimicrobial agents, anti— acne agents, antioxidant agents, cooling , soothing agents, wound g agents, anti-inflammatory—agents, penetration enhancers, permeation enhancers, anti—oxidants, anti-aging , anti-wrinkle agents, skin whitening or bleaching agents, ultraviolet (UV) light absorbing or scattering agents, skin depigmentation agents, regenerative agents, scar healing agents, dyes or coloring , deodorizing agents, fragrances, heratolytic agent, and any combinations thereof. in some embodiments, the active agent can he a heratolytic agent. {653111} in some embodiments, the active agent is an anti—inflammatory agent, As used herein the term “antininflammatory agent” refers to a compound (including its analogs, derivatives, prodrugs and pharmaceutically salts) which can be used to treat inflammation or inflammation related e or disorder.

Exemplary anti~inilammatory agents include, but are not d to, the known steroidal antiwintlammatory and non-steroidal anti=inflammatory drugs (NSAiDs). ary steroidal anti—inflammatory agents include but are not limited to 2i— acetoxypregnenolone, alclometasone, algestone, amcinonide, heclomethasone, betarnethasone, hudesonide, chloroprednisone, clobetasol, tansone, it) clocortolone, cloprednol, osterone, cortisone, cortivazol, deflazacort, desonide, desoxiinctasone, dexamethasone, diflorasone, diflucortolone, ditluprednate, enoxolone, fluazacort, flucloronide, finmethasone t‘lunisolide, fluocinolone acetonide, nonide, rtin butyl, fluocortolone, iluorometholone, flnperolone e, iluprednidene e, iluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, haiometasone, halopredone acetate, hydrocortarnatc, hydrocortisone, loteprednol etahonate, mazipredone, medrysone, nisone, methylprednisolone, mometasone furcate, paramethosone, prednicarbate, prednisolone, prednisolone 25udiethylamino—acetate, prednisolone sodium phosphate, prednisonc, prednival, prednylidenc, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triatncinolone benetonide, triamcinolone hexacetonide, derivatives thereof and mixtures f, Exemplary nonsteroidal anti—inflammatory agents include but are not limited to COX tors (CQXJ or COX nonspecific inhibitors) and selective COX~2 inhibitors. Exemplary CGX inhibitors include but are not limited to salicylic acid derivatives such as aspirin, sodium salicylate, choline magnesium icylate, salicylate, diilunisal, sulfasalazine and olsalazine; minophenol derivatives such as acetaminophen; indole and indene acetic acids such as indomethacin and sulindac; heteroaryl acetic acids such as tolmetin, dicofenac and ketorolac; arylpropionic 3@ acids such as ibuprofen, naproxen, profen, ketoprofcn, fenoproten and oxaprozin; anthranilic acids (fenamates) such as mefenamic acid and meloxicam; enolic acids such as the oxican’is (piroxicam, meloxicam); nes such as nabumetone; derivatives thereof and mixtures f. Exemplary CQX—Z inhibitors e but are not limited to diarylsubstituted furanones such as xib; diaryl—substituted pyrazoles' such as celecoxib; indole acetic acids such as etodolac and sulfonanilides such as nimesulide; tives thereof and mixtures thereof. {£33112} in some embodiments, the active agent is an anti~aging agent. As used herein, the term “anti—aging agent” means a compound or ition that inhibits or reduces signs of aging, such as wrinkles, fine lines, and other stations of photodamage, Examples of anti—aging agentsinclude, but are not limited to, ilavonoids such as quercetin, hesperidin, quercitrin, rutin, tangeritin, and epicatechin; Cletl; nic sunscreens such as tianium dioxide and zinc oxide; organic sunscreens such as octyl- methyl cinnamates and derivatives thereof; retinoids; vitamins such as vitamin E, vitamin A, vitamin C (ascorbic l5 acid), vitamin B, and derivatives thereof such as n E acetate, n C palinitate, and the like; antioxidants including alpha hydroxy acid such as glycolic acid, citric acid, lactic acid, malic acid, mandolic acid, ascorbic acid, alphahydroxybutyric acid, alpha— hydroxyisobutyric acid, alphanhydroxyisocaproic acid, atrrolactic acid, alpha- hydroxyisovaleric acid, ethyl pyruvate, galacturonic acid, glucopehtonie acid, glucopheptono l,4~laetone, gluconic acid, gluconolactone, glucuronic acid, glucurronolactone, glycolic acid, isopropyi pyruvate, methyl te, mucic acid, pyruvia acid, saccharic acid, saccaric acid l,4—lactone, tartaric acid, and tartronic acid; beta hydroxy acids such as beta~ hydroxybutyric acid, beta-phenyl-lactic acid, beta-phenylpyruvic acid; cal extracts such as green tea, soy, milk thistle, algae, aloe, angelica, bitter orange, coffee, goldthread, grapefruit, hoellen, honeysuckle, Job’s tears, lithosperrnum, mulberry, peony, puerarua, rice, safflower, and mixtures thereof. {titlll3l in some embodiments, the active agent is an ultraviolet (UV) light absorbing or scattering agent. Ultraviolet light absorbing agents include, for example, ultraviolet absorber of c acid system such as para-aminobenzoie acid (hereinafter, abbreviated as PARA), PARA monoglycerin ester, N,N- dipropoxy PABA ethyl ester, N,Nudiethoxy PABA ethyl ester, N,N—dimethyl PABA ethyl ester, methyl PARA butyl ester, and N,N—dirnethyi PARA methyl ester and the like; ultraviolet absorber of anthranilic acid system such as honiomenthyl-N—acetyl anthranilate and the like; ultraviolet absorber of salicylic L)! acid system such as arnyl salicylate, menthyl salicylate, homomenthyl late, octyl salicylate, phenyl salicylate, benzyl salicylate, ropanol phenyl salicylate and the like; iolet absorber of cinnamic acid system such as octyl ate, ethyl-ll-isopropyl atc, methylw2,5r=diisopropyl cinnainate, ethyl: 2,4wdiisopropyl cinnamate, methyl—2,4-—diisopropyl cinnamate, propylmp—methoxy cinnamate, isopropyl—p-methoxy cinnamate, isoamyl~p~methoxy cinnamate, octyl~ oxy nate(2—cthylhexyl-p~rnethoxy cinnamate), 2wetlioxycthylwp— methoxy cinnamate, cyclohexyl-p~rnethoxy' cinnamate, ethyl~o—cyano~l3—phenyi cinnamate, 2—ethylhexyl-o—cyano—B—phenyl cinnamate, glyccryi mono-2— ethylhexanoyl-dipara—methoxy cinnaniate, methyl.- bis(trimethylsiloxane)silylisopentyl trirnethoxy cinnamate and the like; 3—(4’- metliyibenzylidene)—d,l-camphor; 3—benzylidene—d,l-earnphor; urocanic acid, urocanic acid ethyl ester; Z—phenyl—5~methylhenzoxazole; ydroxy methylphenylbenzotriazole; 2—(23vhydroxyw5‘=t—octylphenyl)benzotriazole; 2—(2’— hydroxy—S’-niethylphenylhenzotriazolef, dibenzaladine; dianisoylmethane, 4~ methoxy—4’wt-hutyldibenzoylmethane; 5~t3,3~dirnethyl—2~norbornylidene)= pentane-fZ—one; dirnorpholinopyridazinone; and combinations thereof. Ultraviolet light scattering agents include, for example, powders such as titanium oxide, particulate titanium oxide, zinc oxide, particulate Zinc oxide, ferric oxides particulate ferric oxide, ceric oxide and the like.

Ex) kh Etitlildl in some embodiments, the active agent is an anticwrinlrle agent, e.g., a ological anti-wrinkle agent. Anti—wrinkle agents include, without limitations, flavonoids such as quercetin, hesperidin, quercitrin, rutin, tangeritin, and epicatechin; CleO; Vitamin C; hydroxy acids including C2 ~C30 alpha- hydroxy acids such as glycolic acid, lactic acid, Z—hydroxy butanoic acid, malic acid, citric acid tartaric acid, alpha—hydroxyethanoic acid, hydroxycaprylic acid and the like; beta hydroxy acids including salicylic acid and polyhydroxy acids .

WO 14666 including gluconolactone (G4); and mixtures of these acids. Further anti- wrinkleagentsinclude retinoic acid and gamma—linolenic acid, lfltlllSl in some embodiments, the active agent is a skin ing or bleaching agent. Skin whitening and bleaching agentsinclude hydrogen peroxide, zinc peroxide, sodium peroxide, hydroquinone, 4—isopropylcatechol, hydroquinone monobenzyl ether, kojic acid; lactic acid; ascorhyl acid and derivatives such as magnesium ascorbyl phosphate; arhutin; and licorice root, Sunless tanning actives include dihydroxyacetone (DEA); glyceryl de; tyrosine and tyrosine derivatives such as malyltyrosine, ne glucosinate, and l0 ethyl tyrosine; phospho—DGPA, indolcs and derivatives; and mixtures f.

Other shin whitening agents include sugar amines, such as glucosamine, yl glucosamine, glucosamine sulfate, matmosamine, N—acetyl mannosamine, galactosamine, N-acetyl galactosamine, their isomers (cg, stereoisomers), and their salts (cg, HCl salt); and N-acyl amino acid compounds, such as Nnaeyl l5 phenylalanine, Nwacyl tyrosine, their isomers, including their 1) and L isomers, salts, derivatives, and mixtures thereof. An example of a suitable Nmacyl amino acid is N— undecylenoyl-L-phenylalanine. llltlllél ln some ments, the active agent is a skin depigmentation agent.

Examples of suitahle depigmentationagentsinclude, but are not limited to, soy extract; soy isotlavones; retinoids such as retinol; kojic acid; kojic dipalmitate; hydroquinone ; arhutin; transexamic acid; vitamins such as niacin and vitamin C; azelaic acid; linolenic acid and linoleic acid; placcrtia; licorice; and ts such as chamomile and green tea; and salts and prodrugs thereof. liltll 1.7} in some ments, the active agent is an antioxidant agent. As used herein, the term “antioxidant agent” refers to any le capable of slowing, ng, inhibiting, or preventing the oxidation of other molecules.

Examples of antioxidants include, but are not limited to, hydrophilic antioxidants, lipophilic antioxidants, and mixtures f. Non-limiting examples of hydrophilic idants include chelating agents (eg, metal ors) such as 3Q ethylenediaminetetraacetic acid (EDTA), citrate, ethylene glycol tetraacetic acid (EGTA), l,2—his(o—aminophenoxylethanexNNN’,N’—tetraacetic acid (BAPTA), lene triamine cetic acid (DTPA), mercapto-l—propanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), d-lipoic acid, salicylaldehyde isonicotinoyl hydrazone (Sill), hexyl thioethylamine hydrochloride (ETA), desferrioxamine, salts f, and mixtures thereof. Additional hydrophilie antioxidants include ascorbic acid in C), cysteine, glutathione, dihydrolipoic acid, aptoethane sulfonic acid, 2-mercaptobenzimidazole sulfonic acid, 6=hydroxy=25,7,8—tetramethylchroman—Ewcarboxylic acid, sodium metabisulfite, salts thereof, and mixtures thereof. Non-limiting es of lipophilic antioxidants include vitamin E isomers such as (1-, Ba, v—, and @— tocopherols and on, l3=, ya, and 5~tocotrienols; polyphenols such as 2~tert~hutyl-4~ methyl phenol, Z-tert-hutyl—S—methyl phenol, and 2utert—butyl-6—methyl phenol; butylated hydroxyanisole (BEA) (cg, 2—tert-butyl-4—hydroxyanisole and 3—tert— hutyl—4—hydroxyanisole); hutylhydroxytoloene (Bl-ll"); utylhydroquinone (TBHQ); ascorbyl palmitate; nupropyl gallate; salts thereof; and mixtures thereof. l5 One of skill in the art will appreciate that antioxidants can be classified as primary antioxidants, secondary antioxidants, or metal chelators based upon the mechanisms in which they act. Primary antioxidants quench free radicals which are often the source of oxidative ys, whereas secondary antioxidants on by decomposing the peroxides that are reactive intermediates of the pathways, Metal ors function by sequestering the trace metals that promote free radical development. in some embodiments, the antioxidant agent is resveratrol, Emilia} in some embodiments, the active agent is a wound healing agent. As used herein, theterm ”wound healing agent" means active agents that are effective for promoting natural wound healing processes over days, weeks, or months.

Exemplary wound healing agents include, but are not limited to, proteinaceous growth factors, vascular endothelial growth factors, anti—proliferant agent, antirnicrohials, and anti—inflammatory agents. ltltllltil in some embodiments, the active agent is a soothing agent. As used herein, the term ”soothing agent" means a molecule which helps in reducing the discomfort of the skin and/or scalp, for e by soothing the feelings of itching. Exemplary soothing agents include, but are not limited to, aloe, o oil, green tea extract, hops extract, chamomile extract, colloidai oatmeal, calarnine, cucumber extract, sodium paimate, sodium palm kernelate, hutyrospermurn parkii (i.e., shea butter), menthe piperita (i,e., peppermint) leaf oil, sericin, pyridoxine (a form of n as), retinyi palniitate and/or other forms of vitamin. A, tocopheryl e and/or other forms of vitamin E, latuyi iaurate, hyaluronic acid, aloe harhadensis leafjuice powder, euterpe ea (i.e., acai berry) fruit extract, riboflavin (i.e., vitamin B2), thiamin HCl and/or other forms of vitamin Bi, and/ any combinations thereof. ititiiZtii in some embodiments, the active agent is a cooling agent. As used herein, the term “cooling agent” refers to molecules which provide a sensation of cooling on application. Some exempiary cooling agents include, but are not limited to, WEE-=3; WSa—23; menthol; 3~suhstituted-P—menthanes; N—suhstituted-P- menthaneuEucarboxamides; isopuiegol; 3-»(1— xy)propane-—l,Z—dioi; 3—(ta menthoxy)—2~methylpropane-1,Z—diol; p-menthanew’lfindiol; p—menthane-Sfi—dioi; 6-isopropyi—9-methyl—l,4—dioxaspiro[4,5]decane-2~methanoi; menthyl ate and its alkaline earth metal salts; trimethylcyclohexanol; lh—ethyi—E-isopropyi—S- methylcyciohexanecarhoxamide; Japanese mint oil; peppermint oil; ne; ne giycerol ketal; menthyi lactate; enthoxy)ethan-l~ol; Butl— 2t) xy)propan-—l-ol; 3—(1— menthoxy)butan—i—ol; hylaeetie acid N~ ethylainide; i~menthyi~4-hydroxypentanoate; l- menthyl-3~hydroxyhutyrate; N,2,3—trimethyl-’7—(l—methy1ethyl)—butanarnide; n—ethyl~t~2—c—6 nonadienamide; N,N-dimethyl menthyi succinamide; inenthyl pyrroiidone carhoxylate; and the like. {@121} in some embodiments, the active agent is a coloring agent, As used herein, theterm ”coloring agent" means any substance that canhe employed to produce a desired color. Gen. Such coloring agents are approved for human consumption pursuant an appropriate governmental agency and/or act, such as the Food and Drug Administration (FDA)/Federal Food Drug and Cosmetic Act 3t) (FD&C) in the US or an anaiogous agency of the European Union. For example, oring agent can he a food-grade dye or a take. A ”dye" is a water soluble compound, which is available as a powt‘ler, granule, liquid or other special purpose form. A ”lake" is a water insoluble form of a (lye. Exemplary coloring agents include, but are not limited to, FD&C Blue No, l. (Brilliant Blue), roac Blue No 2 otine), FD&C Green No, 3 (Fast Green), FEé’cC Red No. 3 (Erythroslne), Fl)&C Red No, 4% {Allure Red), FD&C Yellow No. 5 (Tartrazine), FD&C Yellow No. 6 t Yellow), annatto extract, anthocyanis, aronia/redfiuit, beet juice, beet powder, beta—carotene, beta~apo-8* nal, black currant, burnt sugar, xanthin, caramel, carbo medicinalis, carrnine, carmine/beta—carotene, carmine blue, carrninic acid, carrot, carrot oils, ll} chlorophyll, chlorophyllin, cocbineal t, copper—chlorophyll, coppen chlorophyllin, curcumin, curcumin’Cu—cblorophyllin, elderberry, grape, grape skin. extracts, hibiscus, lutein, mixed carotenoids, paprika, naprilta extract, paprika cleoresin, riboflavin, saffron, spinach, stinging nettle, titanium e, ic, and combinations thereof. Preferred coloring agents according to the present invention are FD&C Blue No. l (Brilliant Blue), FDELC Blue No, 2 {lntligotine}, FD&C Green No. 3 (Fast Green), FD&C Red No. 3 (Erythrosine), FD&C Red No. 40 (Allura Red), FD&C Yellow No. 5 {Tartrazine}, FD&C Yellow No. 6 (Sunset Yellow), and any combinations thereof, ltltthZZl in some embodiments; the active agent is a fragrance. Exemplar")r fragrances include; but are not limited to, 2,4—dimethyl—3-cycloltexene~l.~ dehytle; isocyclocitral; menthone; thone; RQMASCQNE® (methyl 2,2-dimetliyl-6—metliylene—l ~cyclohexanecarboxylate)', nerone; terpineol; diliydroterpineol; terpenyl acetate; oibyoroterpenyl acetate; dipentene; eucalyptol; ate; rose oxide; PERYCQROLLEGD ((3)4,8-p—mentliadiene—7—ol); l—p— inentbene—4—ol; (lRS,3RS,4SR)~3-p~mentanyl acetate; (lR,2S,4R)—4,6,6— hyl~bicyelol3 , l , l lbeptan~2=ol; DOREMOX® (tetrahytirernethle- -EH—pyran); cycloliexyl acetate; cyclanol acetate; Fructalate (l,4~ cycloliexane dietliyldicarboxylate); KQUMALACTONE® ,6SR,7ASR)~ perhydro~3,6-dimetltyl—benzo[Blfurana'Z—one); Natactone ((6R)=perliydro-3,6- dimetliyi—benzollfi]tirran-2~one); 2,4,6—trimetltyl~4-plienyl—l,3~dioxane; 2,4,6, trlmetliyl~3~cyclobexene—l wearbaldehyde; (3-3 =metbyl—5-(2,2,3-trimethyl—3— cyclopentcnm 1 —yl)-4—penten-2—ol; (1‘R,E)—2-ethyi—4—(2',2’,3'«trimethyl—E’- cyclopanten-t '—yE)—2—buten~ l ~0E; PQLYSANTQL® ((1 ER,E)~3 ,3~dimathyl—5- (222‘,3’—trimethyl~3i—cyciopentem1’~yl)-4—pentsn~2~oi); fleuramone; PARADESGNE® l—(1R)—cis—3—ox0—2—pentyl-1—cyc§0pentane acetate); U: Veioutene (2,2,5~Trimethy1—5-pantyt~1-cycfiop¢ntan0ne); NIRVANOL® (3,3— dimethyE~5—(2,2,3-trimt:thyis3—cyciopentcn—t —y‘a)—4~penten—2woi); 3~m€thyi~5~ -trimethyi—3~cyc10penten~i—yE)—2—pentan01; damascones; NEOBUTENGNE® (1st”SSS—dimethyl- 1 hexen— 1 ~yE)penten— 1 -one); nectaiactone (( 1 °R)-2—{2~ (4°-methyE-3’—cyciohexen~1'—yl)propyi}cyciopentanone); aipha—ionone; beta» H) fictions; damascenena; ONE® (mixture of t—(Sfi—dimethyi—t— cyclohexen-l—yE)-4np€nten—1-ona and 1-(3,3—dim€thyI-l~cycioh¢xen~l-yfi)—4— penten-t—one); DORINONE® beta (1-(2,6,6~trimethyi~l-cyciohexen—t~y§)—2~ buten— 1 —0ne); RC5MANDOLIDE® ((18,1ER)~{1~(3’,3i—DimethyEv-l'- cyciohaxyDethoxycarbonyflmethyI propanoate); 2—tert—butyi~1—cyctohexyi acetats; LEABANOLGE (1-(2,2,3,6~tatramethyl-cyclohexyi)~3~hexan03); trans~1~(232,6— trimethyl— 1 —cyct0hexyi)=3 —hexan0i; (E)=3—methyi=4-(2,6,6utt'intethyi—2— cyctohexen—l~yi)=3—buten—2—one; terpenyi isobutyrate; LORYSEA® («it-(1,1— dimethyiethyl)- i ~cyciohexyi acetate); S—nigthoxyp=m€nthene; HELVETOLIDE® ((1 SS 1 ‘R)—2-{ l”(3‘,3’=dimethyi—1'-cyci0h€xyi) sthexyj—Z- methyipropyi pmpanoate); para tsrt-butyicyclehexanone; menthenethioi; 1~ mathyl(4—methyl~3~p€meny1)-3ucyciohaxsne— 1 -carhaidehyde; ally} cyclohexylpropionate; cyciohexyi late; Methyl cedryt listens; Vsrdyiate; vetyvemfi; mne; 1-(octahydm-2,3,8,8—tetramathyt~2*naphtaiany];—1- ne; (SRSBRS, 639,1Q—tetramethyi—I~0xaspiroi4fijdeca-3 ,6—diene and the (SRSBSRJQRS) isomsr; 6—ethyE—2,10,1fi~trimethyt~t~0xaspir0[4.5]déca— 3 ,6~diana; l,2,3,5,6,7-hexahydm~1,1,2,3 ,3=pentamsthyl-4—indenene; I—HVERNAI_,® (a mixture of 3=(3,3-dimethyEindanyi)pmpanai and 3~(i,1~ dimethyE-S~indanyi)pmpanal); Rhubofix® (3’,4=dimethyi- tricyclcsEél 1 .0(2,7)]undec—ftuene—Q—spiro»2’—0xirane); 9/10—ethyldiene—3— oxatricycioi623 .G(2,7)}undecam€; OD® (perhydro—S55,8A—trimethyl—2~ atcnyi acatata); octa‘iynol; CETALOX® ‘(dodecahydro-Saaéfifia— tetramethyi-naphthoiz,ivbjfuran); io{5.2.i.0(2,6)]dee—3—en—8-yi acetate and trieyeio[5.2.1.9(2,6)}decen—8~yi acetate as weii as trieyeioiSQl.0(2,6)}dee—3— en-g-yi propanoate and tricycioESQ.i.0(2,6)jdee=4—en—8—yi propanoate; camphor; hemeei; isehornyi acetate; 8—iseprepyi~6~methyi~hieyelo{2.2.2}octene—2— earhaidehyde; camphopinene; cedramher (3-rnethexy~2,6,6,8~tetramethyi~ tricyeioE5.3.i.0( ndecane); cedrene; cedrenei; ; FLQREX® (mixture 0f g-ethyiidene-E~oxatrieyeloifi2. i .G{2,7)]undeean—4wone and iti—ethyiidene-B— oxatricyeieidl i .O(2,7)]undecan~4~one); 3~rnethoxy-7,7-dimethyi—i yiene— bieyeioidfi. i ideeane; CEDROXYDE® (trimethyi-i 3—oxabieycio—[i 0.1 .0]-trideea= it) 4,8—diene); Anthrettuiide LG {(E)hexadecen—i6—eiide); HABANGLEDE® (pentadeeenoiide); museenone hyi—(d/E)weyciopentadeeenene); museone; EXALTOLIDE® {pentadeeaneiide}; EXALTONE® (eyeiepentadecanune); (iu ethoxyethoxy)eyeiodedeeane; Astrotene; LILIALQE; rosinei; and the iike. {$9123} in some embodiments, the active agent is an ngai agent.

Exampiary antifungai agents are described elsewhere in the disciaoure. As used herein, the terms “fungus” or “tungi” include a variety of nueieated, spore~hearing sms which are devoid of ehiorophyii. Exampies ineiude yeasts, miidews, molds, rusts, and mushrooms. es of fungi inetude, but are not iirnited to iilusfiimigates, Aspergiliusflavus, Aspergz‘lius nidulans, Candida aibicans, Candida glabrata, Candida gaiiliermandii, Candida krusei, Candida lusiiam’ae, Candida parapsilosis, Candida tropicalis, Crypiacaccus neafurmans, Issatchenkia orientaiis, Coccidiaides, Paraeeccia’ioides, Histopiasma, myees, Trichophyiaa rubrymfind Neurespora crassa. in some embodiments, fungus is of the genus ezia (e.g., M. far/id, M. pachydermatis, M. globasa, M. restricta, M. siaafiiae, M. z’aiis, M. nana, M. yamataensis, M. dermatis, and M. obtuse). in one embodiment, the fungus is Trichaphymn rubrum. {@124} in same embodiments, the active agent is an antibacterial agent, Exempiary acterial agents are described elsewhere in the disclosure. 3t} EtifiiZSi in some embedimens, the active agent is an anthsearring agent. As used herein, an “anti~searring agent” refers te any agent which inhibit fibrosis or scarring. Usefiil carring agents can inhibit one or more aspect of the fibrosis process. For example, in certain ments, the anti-scarring agent inhibits ation; collagen production in, or release from, cells; and/or is an anti— infective or antifnngal agent. in some embodiments, the anti-scarring agent is selected from the group consisting of (—i—arctigenin, 6. The device of claim l or claim 2 wherein the anti—scarring agent is selected from an angiogenesis inhibitor, a 5-HT inhibitor, a beta 1 integrin antagonist, a beta tnbnlin inhibitor, a bisphosphonate compound selected from onate and an analogue or derivative thereof, a blocker of enzyme production in Hepatitis C, a bone if) mineralization er, a Bruten’s tyrosine kinase inhibitor, a calcinenrin inhibitor, a calcium l blocker, a Caivi kinase ii inhibitor, a caspase 3 inhibitor, a cathepsin B inhibitor, a cathepsin K. inhibitor, a eathepsin L inhibitor, a CBl/CBZ receptor agonist, a CC chemokine receptor antagonist, a CD40 antagonist, a cell cycle inhibitor, a cell cycle inhibitor, a chernokine receptor antagonist, a chymase inhibitor, a clotting factor, a collagenase nist, a coal integrin inhibitor, a CXCR antagonist, a cyclic GM? agonist, a cyclin dependent kinase inhibitor, a cyclooxygenase l inhibitor, a D2 ne receptor antagonist, a DliFR inhibitor, a diuretic, a DNA alkylating agent, a DNA methylation inhibitor, a DNA methylation promoter, a DNA methylation er, a DNA synthesis inhibitor, a DNA topoisomerase inhibitor, a ne antagonist, a famesyltransl’erase inhibitor, a tarnexyl transferase inhibitor, a ogen antagonist, a G protein agonist, a glyeosylation inhibitor, a heat shock protein 9% antagonist, a histamine or antagonist, a e dcacetylase inhibitor, a histone deacetylase inhibitor, a JAKZ inhibitor, a JAK3 enzyme inhibitor, a INK inhibitor, a kinase tor, a n antagonist, a leukotriene inhibitor and antagonist, a iysyi hydrolase inhibitor, a MAP kinase inhibitor, a matrix metalloproteinase inhibitor, a ubnie inhibitor, a microtnbuie inhibitor, a muscarinic receptor inhibitor, a neurokinin antagonist, a nitric oxide agonist, a nitric oxide synthase inhibitor, a NO synthase inhibitor, a norepinephrine renptake 39 inhibitor, a NSAID agent, a p38 MAP ltinase inhibitor, a palmitoyi-protein thioesterase inhibitor, a PDGF receptor kinase inhibitor, a peptidyiglycine alpha- hydroxyiating ygenase inhibitor, a peptidyhproiyi cis/trans isomerase tor, a Peptidyl—Prolyi Cis/Trans isomerase inhibitor, a peroxisome proiiferator—activated receptor (PPAR) agonist, a pesticide, a phosphatase inhibitor, a phosphotiiesterase tor, a PKC inhibitor, a PKC inhibitor, a piateiet activating factor antagonist, a platelet aggregation inhibitor, a poiyntorphonnclear neutrophii inhibitor, a proiyl hydroxyiase inhibitor, a giandin inhibitor, a protein synthesis inhibitor, 3. protein tyrosine kinase inhibitor, a pnrineoreceptor P2X antagonist, a pyruvate dehydrogenase activator, a Raf kinase inhibitor, a RAR/RXT antagonist, a reducing agent, a retinoic acid receptor nist, a retinoio acid or antagonist, a selective serotonin reuptake tor, a serine protease inhibitor, a serotonin receptor inhibitor, a sheddase inhibitor, a sodium channei tor, a steroid, a steroid, a stromelysin inhibitor, a superoxide anion generator, a "PACE inhibitor, a teiomerase inhibitor, a TGIF beta inhibitor, a thromboxane A2 receptor inhibitor, a TNF— alpha nist, a Toii receptor inhibitor, a tryptase inhibitor, a tobniin antagonist, a tumor necrosis factor antagonist, a ne kinase inhibitor, a VEGF tor, a vitamin D receptor agonist, ampiciiiin sodium sait, an acetyichoiinesterase inhibitor, an actin polymerization and stabiiization promoter, an adenyiate e agonist, an A LK-S receptor antagonist, an alpha adrenergic receptor antagonist, an androgen inhibitor, an anesthetic compound, an ensin ii receptor t, an antibiotic seiected from the group consisting of apigenin, an anti-coagniant, an anti-emetic- agent, an anti—inflammatory compound, an antimetabolite and antineoplastic agent, an anti—microbial agent, an anti-microbiai agent, an anti- neoplastic agent, an antioxidant, an antinproliferative agent, an anti-psychotic compound, an anti—spasmodic agent, an antithrombotic agent, an anti-viral agent, an apoptosis activator, an apoptosis tor, an sis antagonist, an aromatase inhibitor, an AXOR12 agonist, an eiastase inhibitor, an eiF-Za inhibitor, an elongation factor-i. alpha inhibitor, an endothetiai growth factor antagonist, an endotheiial growth factor receptor itinase inhibitor, an endotoxin antagonist, an epothiione and tnbuiin binder, an estrogen agonist, an estrogen receptor antagonist, an FGF inhibitor, an FCiF receptor kinase inhibitor, an FLT-=3 hinase inhibitor, an FXR antagonist, an HMGCoA reductase inhibitor, an HMGCoA reductase inhibitor, an lCAM tor, an ii, an lL—2 inhibitor, an irnmttnosuppressant, an tor of type lll receptor tyrosine kinase, an inosine monophosphate inhibitor, an interleukin antagonist, an intracellular calcium flux U1 tor, an intracellular calcium flux inhibitor, an intracellular m influx tor, an irreversible inhibitor of enzyme methionine arninopeptidase type 2, an isozyme selective delta n kinase C inhibitor, an MCF-CCRQ, inhibitor, an MEKl/MEK 2 inhibitor, an MIF inhibitor, an mTOR inhibitor, an m’l‘OR kinase inhibitor, an NF kappa 3 inhibitor, an ornithine decarboxylase inhibitor, an S— ll) adenosyl—L-homocystcine hydrolase inhibitor, an SDF-l antagonist, an SRC tor, an Syk kinase inhibitor, an d—glucosidase inhibitor, an tntegrin antagonist, and a irnrnuno~ modulator selected from Bay ll—7085, and lRAK antagonist, lCE, idazo‘xan hydrochloride, protein itinase B inhibitor, protein hinase C stimulant, purine nucleoside analogue, purornycin, ible inhibitor of ErbBl and ErbBi’Z, ribonucleoside triphosphate reductase inhibitor, an“)i combination thereof, in some embodiments, the anti-scarring agent can be selected from ZD~6474, AP-ZSS’ZS, synthadotin, S-G885, ne, ixabepilone, lDN-SEQG, SB~2723005, ART—Sig, combretastatin, anecortavc acetate, SB- 7l5992, ternsirolirnus, adalirnnrnab, erucylphosphocholine, alphastatin, etanercept, humicade, gefitinib, isotretinoin, radicicol, clobetasol propionate, homoharringtonine, triohostatin A, brefeldin A, thapsigargin, dolastatin, cerivastatin, jasplahinolide, herbimycin A, pirfenidone, Vinorelbine, l7-DMAG, imus, loteprednol etabonate, juglone, prednisolone, purornycin, E, cladribine, inannose-o-phosphate, 5- azacytidine, Ly33353l (ruboxistaurin), and sirnvastatin. maize} ln some embodiments, the active agent is a sltin regenerating agent.

Some skin regenerating agents can act as anti-scarring agents. {@127} in some embodiments, the drug carrier comprises an additional anti— acne agent, in some embodiments, the onal anti—acne agent can be selected 3t} from the group consisting of acetretin, adapalen.e(s}, alitretinoin, alpha- or beta~ liydroxy acids, antibiotics, antimicrobial peptides, antimicrobials, azelaic acid, benzoyl peroxide, bexarotene, bile salts, biofilm inhibitors, clindamycin, erythroniycin, etretinate, glycolic acid, isotretinoin, heratolytic agents, lactic acid, lipoic acid, N—acetylcystein, natural anti-acne agents, octopirox, yethanol, phenoxypropanol, pyruvic acid, resorcinol, retinoic acid, retinoidts), salicylic acid, sebostats, sodium snifacetamide, oiactone, sulfur, sulfur containing D— or L—amino acids, tazarotene, tea tree oil, tretinoin, triclosan, urea, and any combinations thereof. {00123} The drug carrier disclosed herein can comprise any amount of the APE (e.g., DART or other agent). For example, the drug carrier can se about l0 0,0l% to about 99% tw/w) of the APl. For example, the particle can se between about 0.0l% to about 20% (w/w) of the APE. in some ments, the APl comprises greater than 1 % (w/w), greater than 5% (w/w), greater than 10% (wl'w), greater than 15% (w/w), greater than 20% (w/w), greater than 25% (w/w), r than 30% (w/w), greater than 35% (w/w), greater than 40% (w/w), greater than 45% (w/w), greater than 50% (WI/W), greater than 55% (w/w), greater than 60% (w/w), greater than 65% (w/w), greater than 70% (w/w), greater than 75% (w/w), greater than 80% (w/w), r than 85% (w/w), greater than 90% (Wt/Vii), or greater than 95% (w/w) of the total weight of the drug carrier. in some embodiments, the content of Al’l in the drug carrier can range from about 75% to about 97% (MW). in some other embodiments, the content of APl in the drug carrier can range from about 3% to about 25% (w/w). [lllll29l A lipid for use in the drug carriers or formulations disclosed herein can be selected from the group consisting of fatty acids, fatty alcohols, glycerolipids (cg, monoglyceridcs, erides, and triglycerides), olipids, glycerophospholipids, olipids, sterol lipids, prenol , saccharolipids, polyketides, and any combination thereof. in some embodiments, the lipid can be selected from the group consisting of l,3-Propanediol Dicaprylate/Dicaprate; i0— undecenoic acid; l~dotriacontanoh l—heptacosanol; l—nonacosanoi; Z-ethyl hexanol; Androstanes; Arachidic acid; Arachidonie acid; arachidyl alcohol; Behenic acid; behenyi alcohol; Capmul MCM C10; Capric acid; capric alcohol; capryl alcohol; Caprylic acid; Caprylic/Capric Acid Ester of Saturated Fatty Alcohol Cl2—Cl8; Caprylic/Capric Triglyceride; Caprylic/Capric 'l‘iiglyccridc; Ccramidc phosphorylcholihc (Sphingomyclin, SPH); Ceramide phosphowlethanolamiae (Sphingomyclin; Ccr—PE); dc phosphorylglyccrol; Caroplaslic acid; Ccrolic acid; Carotic acid; ccryl alcohol; (J: Cctearyl alcohol; Cctclhdll; cclyl alcohol; Cholancs; tancs; cholesterol; cis-l lacicoscnoic acid; cis-l l—octadcccnoic acid; cis—l3-docoscnoic acid; cliiylyl alcohol; cocnzymc Q10 (CleG); Dihomoay-linolcnic; Docosahcxaciioic acid; egg lecithin; Eicosapcntacnoic acid; Eicoscnoic acid; Elaidic acid; claidolinolchyl alcohol; claidolinolcyl alcohol; elaidyl alcohol; Emcic acid; crucyl alcohol; lll cs; Ethylene glycol distcaralc (EGDS); chdic acid; gcddyl l; glycerol dislcaralc (type l) E? (Precirol All) 5); Glyccrol 'l‘i‘icaprylatc/Capralc; Glycerol Tricaprylate/Capralc (CAPTEX® 355 ElVNF); glyccryl monocapi’ylatc (Capmul MClvl C8 EP); Glyccryl Triacctatc; Glyccryl Tricapiylatc; Glyceryl Tricaprylalc/Capralchauralc; Glyceryl Tricapiylatc/Tiicapralc; glyceiyl l5 mitalc (Tripalmitin); llcnatriaconlylic acid; lleiicicosyl alcohol; llcncicosylic acid; Hectacosylic acid; llcptadccanoic acid; llcptadccyl alcohol; l-lcxati’iacontylic acid; isostcaric acid; isoslcaryl alcohol; Lacceroic acid; Laurie acid; Lauryl alcohol; Ligncccric acid; ligaoceryl alcohol; Linoelaidic acid; Linolcic acid; nyl alcohol; linolcyl l; Margaric acid; lvlcad; Melissic '20 acid; mclissyl alcohol; ic acid; mohlanyl l; myiicyl alcohol; Myristic acid; Myrisloleic acid; Myristyl alcohol; acodccanoic acid; heoheptanoic acid; neonooanoic acid; Naivonic; Nonacosylic acid; Nonadccyl alcohol; cylic acid; Nonadccylic acid; Oleic acid; olcyl alcohol; Palmitic acid; olcic acid; palmitolcyl alcohol; Pclargoaic acid; pclargoaic alcohol; Pcntacosylic acid; Pentadccyl alcohol; ecylic acid; atidic acid (phosphalida‘ic, PA); Phosphatidylcholinc (lecithin; PC); Phosphalidylcihanolaminc (cephalic, PE); Phosphatidylinosilol (Pl); Fhosphalidyliiiositol bisphocphalc (PlPZ); Phosphatidylihositol ahosphatc (Pll’); Phosphalidylinositol tripliosphatc (PlPIS); Phosphalidylscrinc (PS); polyglyccryl— 6—distcaratc; Pregnancs; Propylene Glycol ale; Propylcnc Glycol Dicapwlocapratc; Propylenc Glycol ylocapratc; Psyllic acid; icciaoleaic acid; ieyi aicohol; Sapienic acid; soy lecithin; Stearic acid; Stearidonic; stearyl alcohol; Tricosyiic acid; Tridecyl aicohoi; Tridecyiic acid; 'ilriolein; Undecyi aicohoi; undecylenic acid; Undecylic acid; Vaccenic acid; o—Linoienic acid; y-Linoienic acid; a fatty acid salt of iD—undecenoic acid, adapaiene, arachidic acid, arachidonic acid, behenic acid, butyric acid, capric acid, capryiic acid, ccrotic acid, cis-l i-eicosenoic acid, cisnil—oetadecenoic acid, cis—iS- docosenoic acid, docosahexaenoic acid, eicosapcntaenoic acid, elaidic acid, erucic acid, heneicosylic acid, osylic acid, heptadecanoic acid, aric acid, iauric acid, lignoceric acid, iinoeiaidic acid, linoieic acid, montanic acid, myristic it) acid, myristoieic acid, neodecanoic acid, neoheptanoic acid, neononanoic acid, nonadecyiic acid, oieic acid, paimitic acid, palmitoieic acid, onic acid, pentacosyiic acid, pentadecyiic acid, recinoieaic acid (eg. zinc recinoleate), ic acid, stearic acid, tricosylic acid, tridecylic acid, undecyienic acid, iic acid, vaecenic acid, vaieric acid, a—iinoienic acid, or y—linoienic acid; i5 paraffin; and any combinations thereof. in some embodiments, the iipid can be a fatty acid comprising ii. or fewer carbons. For example the fatty acid can comprise 6, 7, 8, 9, it), or ii carbons. {tittiStt} Without wishing to be bound by a theory, it is heiieved that fatty acid salts can he also used in the particles to potentiate antiubacteriai activity, e.g., anti- 29 acne activity and provide stability in compositions sing said drug carriers, Accordingly, in some embodiments, the lipid is a fatty acid sait. Without limitations, the fatty acid salt can be selected from the group consisting of zinc, sodium, potassium, lithium, ammonium, copper, caicium, magnesium, strontium, ese, and ations thereof. The drug carrier can comprise any amount of the lipid component. For example, the drug carrier can comprise n about 0.0i% to about 99% (w/w) of the iipid component, in some embodiments, the lipid component comprises greater than 0.1% (w/w), greater than 9.5% (w/w), greater than iii/a (w/w), greater than 2% (w/w), r than 3% (w/w), greater than % (w/w), greater than 5% (w/w), greater than 6% (w/w), r than 7% {w/w), greater than 8% (w/w), r than 9% (w/w), greater than i0% (”w/w), greater than 11% (w/w), greater than i292) (w/w), greater than 13% (w/w), greater than 14% (w/w), greater than 15% (w/w), greater than 16% (w/w), greater than 17% (w/w), greater than 18% (w/w), greater than 19% (w/w), r than 20% {w/w), greater than 25% (w/w), greater than 30% (”w/w), greater than 35% (w/w), greater than 40% (w/w), greater than 45% (w/w), or greater than 50% (w/w) of the LA totai weight of the drug carrier, Typicaiiy, the t of the 1ipid component in the drug carriers are in the range of about 12-25% (w/w). {001311 Ratio of the active agent (e,g., DART or other anti-bacteriai agent) to the totai iipid ent of the coating 1ayer can be any desired ratio. For e, ratio of the active agent to the totai 1ipid component can range from about 100:1 to about 1:100. in some embodiments, the ratio of the active agent to the totai 1ipid component can range from about 75:1 to about 1:75, from about 50:1 to about 1:50, from about 25:1 to about 1:25, from about 20:1 to about 1:20, from about 15:1 to about 1:15, from about 5:1 to about 1:5, or from about 25:1 to about 1:5. in some embodiments, the ratio of the active agent to the totai iipid component is about 30:1, about 25:1, about 20:1, about 15:1, about 10:1, about :1, or about 1:1, The ratio can be based on weight, mass, or moies. {001321 Thickness of the coating layer can range from nanometera to miiiimeters, For exampie, the coating 1ayer thickness can. range from about 1 nm to about 5000 nm, from about 5 nm to about 2500 nm, from about 10 nm to about 2000 nm, from about 50 nm to about 1500 nm, from about 20 nm to about 1000 run, from about 1 nm to about 1000 run, from about 1 nm to about 500 nm, from about 1 nm to about 250 nrn, from about 1 nm to about 200 run, from about 1 nm to about 150 um, from about 1 nm to about 100 nm, from about 2 nm to about 50 nm, or from about 5 nm to about 25 nm. 1001331 in some embodiments, the drug carrier can comprise two or more (cg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) iipids, ie., the carrier can comprise a first iipid and a second lipid, For exampie, the coating 1ayer can comprise a second iipid that is different from the first iipid. 1001341 ary proteins for use in the drug carriers or tonnuiations disciosed herein can include, but are not ed to, Actin, Albumin, Amaranth Protein, Ammonium Hydroiyzed Animal Protein, Animai protein, Barley Protein, Brazil Nut Protein, , Collagen, Collagen protein hydrolyzed, Conehiolin Protein, corn protein, Cottonseed Protein, Elastin, Extensin, Pibroin, eetin, Fish Protein, Gadidae Protein, Gelatin, Glutein, Glyeoproteins, Hazelnut Protein, Hemoglobin, l-lemp Seed n, l-loney Protein, Hydrolyzed Aetin, Hydrolyzed Amaranth Protein, Hydrolyzed animal protein, Hydrolyzed Barley n, Hydrolyzed Brazil Nut Protein, Hydrolyzed Conehiolin n, Hydrolyzed eorn protein, l-lydrolyzed Cottonseed Protein, Hydrolyzed n, Hydrolyzed Extensin, Hydrolyzed Pibroin, Hydrolyzed Pibroneetin, Hydrolyzed Fish Protein, yzed Gadidae Protein, Hydrolyzed Gadidae Protein, Hydrolyzed Gelatin, l0 Hydrolyzed Hair ‘lieratin, llydrol.yzed Hazelnut, Hydrolyzed Hazelnut Protein, Hydrolyzed Hemoglobin, Hydrolyzed Hemp Seed Protein, Hydrolyzed Honey Protein, Hydrolyzed Keratin, yzed Lupine n, Hydrolyzed Maple re Protein, liydrolyzed Milk Protein, Hydrolyzed Oat Protein, Hydrolyzed Pea n, yzed Potato Protein, l-lydrolyzed Reticulin, Hydrolyzed Royal Jelly Protein, Hydrolyzed Serioin, Hydrolyzed Serum Protein, Hydrolyzed Sesame Protein, Hydrolyzed Soy Protein, Hydrolyzed Soymilk Protein, Hydrolyzed Spinal Protein, Hydrolyzed Spongin, Hydrolyzed Sweet Almond Protein, Hydrolyzed Vegetable Protein, Hydrolyzed Wheat , Hydrolyzed Wheat Protein, Hydrolyzed Whey Protein, Hydrolyzed Yeast Protein, l-lydrolyzed 2% Yogurt Protein, l-lydrolyzed Zein, lntegrin, Jojoba protein i-lP, Hydrolyzed, keratin, Lupine n, Maple Sycamore Protein, MEAlZIHydrolyzed Collagen, MEAEl-lydrolyzed Silk, Milk Protein, Myosin, Oat Protein, Pea Protein, polylysine, Potato Protein, Reticulin, Pie-e Quat, Royal. Jelly n, Seriein, Serum Protein, Sesame Protein, Silk powder, Soditun Hydrolyzed , Soy Protein, Soy Rice Peptides, Soymilk Protein, Spinal Protein, Spongin, Sweet Almond Protein, Vegetable Protein, Wheat Gluten, Whey Protein, Yeast Protein, Yogurt Protein, Zein, and Zine Hydrolyzed Collagen. {553135} in some embodiments, the protein is an albumin. The albumin can be a naturally occurring albumin, an albumin related protein or a variant thereof such 3t} as a natural or engineered variant. Variants include polymorphisms, fragments such as s and subdomains, fragments and/or fusion proteins, An albumin can comprise the sequence of an aibumin protein obtained from any source, A number of proteins are known to exist within the albumin family. Accordingly, the albumin can comprise the sequence of an albumin derived from one of serum albumin. from African clawed frog (e. g., see Swissprot accession number P98759= U] l), bovine , see Swissprot accession number P027694), cat (eg., see Swissprot accession number i’49064—t), chicken (cg, see Swissprot accession number PliiiZl-i), chicken ovaibumin (ago see Swissprot ion number PGiGiZ—i), cobra ALB(e.g., see Swissprot accession number Q91134—i), dog (cg, see Swissprot accession number P498221), donkey (e.g., see Swissprot ii) accession number QSXLE4—i), European water frog (e.g., see Swissprot accession number QQYGHt’i-i), blood fluke (e.g., see Swissprot accession number AAL08579 and Q95VB7r-i), Mongolian gerbii (e.g., see Swissprot accession number 0350904 and ), goat (cg, see Swissprot accession number BBVHMQJ and as available from Sigma as product no, A2514 or A4164), guinea pig (e.g., see Swissprot ion number QoWDNQ-i), hamster (see DeMarco et a]. (2007). internationai Journal for Parasitoiogy : 12014208), horse (cg, see Swissprot accession number P35747-i), human (cg, see Swissprot accession number P027684), lian Lungufish (e.g., see rot accession number 1383517), macaque (Rhesus monkey) (ego see Swissprot accession number Q28522—), mouse (e.g., see Swissprot accession number P677244), North American bull frog (egi, see Swissprot accession number P2l847—i), pig (e.g., see Swissprot accession number P()8835—i), pigeon (eg. as defined by Khan et ai, 2002,1ii2. J, Biol. Macromoi, 30(3—4),i7i—8), rabbit (e.g., see Swissprot accession number P490 65—1), rat (e.g., sec Swissprot accession number P02770— 1), saiamander (e.g., see Swissprot accession number Q8UWOS—l.), salmon ALBE (ego see Swissprot ion number ~l), salmon ALBE (as, see Swissprot ion number (2031564), sea lamprey (e.g., see Swissprot accession number (3912744 and 0422794) sheep (e,g,, see Swissprot ion number 4), Sumatran orangutan (e.g., see Swissprot accession number QSNVHS—i), tuatara (e.g,, see Swissprot accession number lelAQ—i), turkey ovaibumin (e.g., see Swissprot ion number O73860~l), Western clawed frog (eg, see Swissprot accession number Q6Di95-1), and includes variants and fragments thereof as defined herein. Many naturally occurring mutant forms of albumin are known. Many are described in Peters, (@926, All About Albumin: Biochemistry, Genetics and Medical Applications, Academic Press, inc, San Diego, California, p.l7G-i.8l), t of which is incorporated herein by reference. The term aibumin also encompasses albumin variants, such as genetically ered forms, mutated forms, and fragments etc. having one or more binding sites that are ous to a binding site unique for one or more albumins as defined above. By analogous binding sites in the context of the it) invention are contemplated structures that are able to compete with each other for binding to one and the same ligand structure in one embodiment, albumin is bovine serum n, egg albumin, hydrolyzed lactalbumin, or lactalburnin, including ts and fragments thereof. in one embodiment, the n is egg albumin. l5 {99136} The protein can comprise between about Gill % to about 99% (w/w) of the drug carrier. in some embodiments, the protein component comprises greater than t),l% (w/w), greater than 0.5% (w/w), greater than l% (w/w), greater than 2% (w/w), greater than 3% (w/w), greater than 4% (w/w), greater than 5% (w/w), greater than 6% (w/w), greater than 7% (w/w), greater than 8% (W/W), greater than 9% (w/w), greater than i0% (w/w), r than 11%. (w/w), greater than l2% (w/w), greater than 13% (w/w), greater than l4% (w/w), r than l5% (w/W), greater than l6% {xv/w), greater than 17% (w/w), greater than l8% (w/w), greater than l9% (w/w), greater than 20% (w/w), greater than 25% (wfw), greater than 30% (w/w), greater than 35% {w/w), greater than 40% (w/w), greater than N (J? 45% (w/w), or greater than 50% (w/w) of the total weight of the drug carriers.

Typically, the t of the n component in the drug rs are in the range of about 1»25% (”w/w), about O.l~l0% (w/w), about 0.56% (w/w), or about l~l 5% (w/w). ititii37} Ratio of the active agent (cg, DART or other anti~bacterial agent) to the protein component can be any desired ratio. For example, ratio of the active agent to the protein component can range from about lOtlzl to about men. in some embodiments, the ratio the active agent to the protein can range from about 100:1 to about 1:1, from about 90:1 to about 10:1, from about 85:1 to about 15:1, from about 80:1 to about 25:13 or from 75:1 to about 50:1. in some emhodimentss the ratio of the active agent to the protein component is about 75:1. The ratio can be based on weight, mass, or moies. {00,1381 Generaiiy, any cationic moiecuie can be used in the drug carriers or ations disciosed herein. As used herein the term “cationic moiecuie”refers to a rnoiecuie that carries a net positive charge. in some embodiments, the cationic moiecuie is a poiyarnine. Exempiary cationic moiecoies inchtde, but are not iirnited to, Putrescine (Butane-1,4adiarnine), Cadaverine newifia tiiarnine), itiine, Sperminefi Cycien (1,437,10—tetrazacyciododecane), Cyciarn (1 ,4,8r1 1 ~Tetraazacyc1otetradecane), Linear Poiyethyieneirnine (Poiyt’iminoethyienefi, Norspermidine , p~Pheny1eneriiarnine (151“ diaminohenzene), Diethyienetriamine (1‘1—(2=arninoethyi)—1 anediamineh osperrnine, Tris(2=arninoethy1)arnine, Hexaniethyienedianiine, ysine (3,6-diaminohexanoic acid). rn—Phenyienediarnine (1,3-diaminohenzene), Diaminopropane (132~Diarninopropane), Ethyienetiiarnine dihydroiodide, and poiyarnine 1) 400 (i’oiyoxyaikyieneamine D 400): {@139} The cationic rnoiecuie can comprise between about 0.01% to about 99% (w/w) of the drug carriert in some embodiments, the cationic nie comprises greater than 0.1% (W/W’L greater than 0.5% (w/w), greater than 1% (wz’w), greater than 2% (Via/W)a r than 3% (w/w), greater than 4% {wt/w)? greater than 5% (w/w), greater than 6% {w/w). greater than 7% {w/w), greater than 8% (w/w). greater than 9% (VJ/W), greater than 10% (w/w), greater than 11% (w/w), greater than 12% (W'l/W'), greater than 13% (w/w), greater than 14% (w/w), greater than 15% (w/w), greater than 16% (Vii/W)? greater than 17% (w/w), greater than 18% (w/w), greater than 19% (w/w), greater than 20% (w/w), greater than % (w/w), greater than 30% (w/w), greater than 35% (w/w), r than 40% (w/w), greater than 45% (w/w}, or greater than 50% (w/w) of the total weight of the drug rs. Typicahy. the content or" the cationic moieeuie in the drug carriers are in the range of about 1~25% (w/w), about 01-10% (w/w), about 0.5— % (W/W), or about 1-1.5% (w/w). {011141.11 Ratio of the active agent (e.g.. DART or other anti—bacteriai agent) to the n component can be any desired ratio. For exampie, ratio of the active UI agent to the protein component can range from about 100:1 to about 1:100. in some embodiments, the ratio the active agent to the protein can range from about 100:1 to about 1:1, from about 90:1 to about 10:1, from about 85:1 to about 15:1, from about 80:1 to about 25:1, or from 75:1 to about 50:1. in some embodiments, the ratio of the active agent to the protein component is about 75: 1. The ratio can be based on weight, mass, or motes. {001.41} Generaiiy, any carbohydrate ie can be used in the drug carriers or forniuiations disciosed herein. in some embodiments, the carbohydrate is a poiysaccharide. Exemplary poiysaccharides inciude ceiiuiose derivatives such as yethyhceiiuiose, hydroxy propyi-methyi—ceiiuiose and carboxymethyh ose; giycosaminogiycans such as hyaiuronic acid, chondroitin suifate, chitin and chitosan; starch derivatives such as starch/hydroxyethyi starch; agarose; and aiginate and combinations thereof. in some embodiments, the carbohydrate can be selected from the group consisting of chitosan and their derivatives, aiginates and their derivatives, puiiuian, their tives {1101142} The carbohydrate can comprise between about 0.01% to about 99% (w/w) of the drug carrier. in some embodiments, the ydrate comprises greater than 0.1% (w/w), greater than. 0.5% (“w/w), greater than 1% (w/w), greater than 2% «(w/w), greater than 3% (w/w), greater than 4% (w/w), greater than 5% (w/w), greater than 6% (w/w), greater than 7% (w/w), greater than 8% (w/w), greater than 9% (w/w), greater than 10% (w/w), greater than 11% (w/w), greater than 12% (WI/‘71,), greater than 13% (W/W), greater than 14% (xv/w), greater than % (w/w), greater than 16% «(w/w), greater than 17% (w/w), r than 18% (w/w), greater than 19% (w/w}, greater than 20% (w/w), r than 25% (w/w), greater than 30% (w/w), greater than 35% (w/w), greater than 40% (w/w}, greater than 45% (w/w), or r than 50% (w/w) of the totai weight of the drug carriers. Typicaliy, the content of the carbohydrate in the drug carriers are in the range of about 1—25% (xv/w), about 0.1~10% (WI/w}, about (LS—5% (xv/w), or about 1«l 5% {w/w). {1111,1433 Ratio of the active agent (cg, DART or other antibacterial agent) to the carbohydrate can be any desired ratio. For example, ratio of the active agent U: to the carbohydrate can range from about 109:1 to about 1:109. in some embodiments, the ratio the active agent to the carbohydrate can range from about 100:1 to about 1:1, from about 96:1 to about 10:1, from about 85:1 to about 15:1,, from about 811:1 to about 25:1, or from 73:1 to about 50:1. in some embodiments, the ratio of the active agent to the carbohydrate is about 75:1. The ratio can be 16 based on weight, mass, or moles. {119144} in some embodiments, the drug carrier r comprises an excipient. in some embodiments, the excipient is a wetting agent. Without limitations, the wetting agent can be selected from alkyl sulfates, e.g. sodium lauryl e, sodium stearyl sulfate, sodium oleyl e and sodium cetyl e, alkyl aryl sulfonates, eg. sodium dodecylbenzene suifonate and diaikyl sodium sulfosuccinates, eg. sodium bis—(2~ethy1hexyl)sulfosuccinate, and most preferably sodium lauryl sulfate. Further examples of the pharmaceutically able g agent include benzethonium chloride, cetylpyridinium chloride, docusatesodium, poloxamer, polysorbate and sorbitan esters, 199145} in some embodiments, the excipient is a stabilizer, cg, a surface stabilizer. Suitable surfacestabilizers can preferably be ed from known organic and inorganic pharmaceutical excipients. Such excipients include various rs, low molecular weight oligomers, natural products, and surfactants with high and low hydrophiiic lipophilic e (111B). Preferred surfacestabilizersinclude ic and ionic surfactants. Two or more surfacestabilizers can be used in combination. Representative examples of estabilizersinclude sodium docusate, cetyl pyridinium chloride, gelatin, , lecithin (phosphatides), dextran, glycerol, gum acacia, cholesterol, tragacanth, stearic acid, benzalkoniuin chloride, calcium stearate, glycerol monostearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan esters, polyoxyetliylene alkyl ethers (cg, maerogoi ethers such as cetomacrogol 1013(1), polyoxyethylene castor oil derivatives? polyoxyethylene sorhitan fatty acid esters (e.g., the commercially available Tweens® such as e.g., Tween 26® and n Ell®lflCl Specialty Chemicals)); polyethylene glycols (e.g., Carbowaxs 335il® and 1450®, and Carhopol 934® (Union Carbide», dodecyl trimethyl ammonium LA bromide, polyoxyethylene steara‘tes, colloidal silicon dioxide? phosphates, sodium dodeeylsulfate, carboxymethylcellulose calcium, ypropyl celluloses (erg, HPC, HPC-SL, and HPC—L), hydroxypropyl methylcellulose (HPMC), earhoxymethylcellulose sodium, methylcellulose? hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethyl~cellulose ale, noncrystalline ll) cellulose, magnesium aluminum silicate, triethanolamine, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), ,3,3-tetramethylhutyl)— phenol polymer with ethylene oxide and formaldehyde (also known as tyloxapol, superione, and triton), poloxamers (e.g., Pluronics F68® and FREQ), which are block copolymers of ethylene oxide and propylene oxide); poloxamines (e.g.., 'l‘etronic 903®, also known as Poloxamine 96869, which is a telrafunctional block mer derived from tial addition of propylene oxide and ethylene oxide to ethylenediamine (BASF Wyandotte Corporation, Parsippany, Nil); a charged phospholipid such as dirnyristoyl phophatidyl glycerol, dioetylsull’osuccinate (BOSS); Tetronic l508® ("l1 lSGS) (BASF Wyandotte Corporation), dialkylesiers of sodium uccinic acid (egg l OT®3 which is a dioctyl ester of sodium sulfosuccinie acid (American Cyanamid)); Duponol P®, which is a sodium lauryl sulfate (DuPont); Tritons X-200®, which is an alkyl aryl polyether sulfonate (Rohm and Haas); Crodestas Ell l0®, which is a mixture of sucrose stearate and sucrose distearate (Croda inc); onylphenoxypolyw(glycidol), also known as Olin-lOG® or Surfactant lO—G®(Olin Chemicals Stamford, CT); Cmdestfis SL- 4il® (Croda, hie); decanoyl.«N-methylglucamide; nudecyl fi—‘Dglucopyranoside; n- decyl {3— D—maltopyranoside; cyl B—D-glucopyranoside; nwdodecyl 3-D— maltoside; heptanoyl—N—methylglucamide; n-heptyl—fi~l3~glucopyranoside; n~ heptyl EH)— thioglucoside; l B-l)»glucopyranoside; nonanoyl-N-= methylglucamide; nmnoyl {3—D glucopyranoside; octanoyl—N»methylglueamide; n- octyl~B—D-glucopyranoside; octyl {i—D— thioglueopyranoside; and the like. Most of these surt‘acestabilizers are ltnovvn pharmaceutical excipients and are described in detail in the Handbook of ceutical Excipients, published jointly by the American Pharmaceutical ation and The Pharmaceutical Society of Great Britain (The Pharmaceutical Press, 1986), content of which is incorporated herein by nce in its entirety. in one embodiment, the exeipient is sodium docusate, {billed} Generally, the drug carriers have an average diameter of from about 5 nm to about 20,000 run. in some embodiments, the drug carriers have an average diameter of from about 5 nm to about 5,006 nm. in some embodiments, the drug carriers have an average er of from about 50 nm to about 2500 nm. in some embodiments, the drug carriers have an average diameter of from about lOO nm to about 2800 nm. in some embodiments, the drug carriers have an average diameter of from about 15G nm to about l7tlilnrn. in some embodiments, the drug carriers have an e diameter of from about 269 nm to about lfifltl nm. in some embodiment, the drug rs have an average diameter of about 260 nm. l5 In one embodiment, the drug carriers have an average diameter of about 30 nm to about l50nm. in some embodiments, the drug carriers have an average diameter of about liltl nm to about 10th run, from about 2th nm to about 806 nm, from about 243% nm to about 790 rim, or from about 3% nm to about 7th run. {bill-47} Generally, the drug carriers disclosed herein can be of any shape or 2% form, e. g., spherical, rod, elliptical, cylindrical, capsule, or disc. {@9148} in some embodiments, the drug carrier can be micro—sized and have a size of about l urn to about lGOG um, in some embodiments, the drug carrier can be ized and have size of about ill rim to about 10th run. in some embodiments, the drug carrier is a micropartiele or a nanoparticle. As used herein, the term “tnieroparticle” refers to a particle having a particle size of about i um to about lOGO urn. As used herein, the term “nanoparticle” refers to particle having a particle size of about (ll nm to about liltlil rim, {96149} it will be understood by one of ordinary sltill in the art that les usually t a distribution of sizes around the indicated “size.” Unless otherwise stated, the terms “drug carrier size” and “particle size” as used herein refer to the mode of a size distribution. of drug carriers or particles, i.e., the value that occurs most ntly in the size distribution. Methods for measuring the drug carrier or le size are known to a d artisan, eg, by dynamic light scattering (such as photocorrelation spectroscopy, laser diffraction, low—angle laser light scattering (LALLS), and medium—angle laser light scattering (MALLS)), light obscuration methods (such as Coulter analysis method), or other techniques (such as rheology, and light or electron microscopy). {$9153} in some embodiments, the drug carrier can be substantially spherical.

What is meant by “substantially spherical” is that the ratio of the lengths of the longest to the shortest perpendicular axes of the drug carrier cross section is less l0 than or equal to about l5, Substantially spherical does not require a line of ry. Further, the drug rs can have surface texturing, such as lines or indentations or protuberances that are small in scale when compared to the overall size of the drug r and still be substantially cal, in some embodiments, the ratio of lengths between the longest and shortest axes of the drug carrier is less l5 than or equal to about l.5, less than or equal to about L45, less than or equal to about 1.4, less than or equal to about 1.35, less than or equal to about 1.30, less than or equal to about 1.25, less than or equal to about LEO, less than or equal to about l.15 less than or equal to about l.l. t wishing to be bound by a theory, surface t is minimized in drug carriers that are substantially spherical, which minimizes the undesirable agglomeration of the drug carriers upon storage, Many crystals or flakes have flat surfaces that can allow large surface contact areas where agglomeration can occur by ionic or non‘ionic interactions. A sphere permits contact over a much smaller area. liltilSll in some embodiments, the drug rs have substantially the same particle size, Drug carriers having a broad size distribution where there are both relatively big and small drug carriers allow for the smaller drug carriers to fill in the gaps between the drug carriers, thereby creating new contact surfaces. A broad size distribution can result in larger spheres by creating many contact opportunities for binding agglomeration. The drug carriers described herein are 3G within a narrow size distribution, thereby minimizing opportunities for contact agglomeration, What is meant by a “narrow size distribution” is a particle size distribution. that has a ratio of the volume diameter of the 90th percentile of the small spherical particles to the volume diameter of the ltlth percentile less than or equal to 5. in some embodiments, the volume diameter of the 90th percentile of the small spherical particles to the volume diameter of the thh percentile is less than or equal. to 4.5, less than or equal to 4, less than or equal to 35,, less than or equal to 3, less than or equal to 205, less than or equal to 2, less than or equal to 1.5, less than or equal to L45, less than or equal to Lilli, less than or equal to L35, less than or equal to l3, less than or equal to 1.25, less than or equal to l.2(l, less than or equal to l.l5, or less than or equal to l.l. {@9152} Geometric Standard Deviation (GED) can also he used to indicate the narrow size distribution. GSD calculations involved determining the" effective cutoff diameter {ECD} at the cumulative less than percentages of 5.9% and 84tl%t GSD is equal to the square root of the ratio of the ECU less than 343794: to ECD less than l5.9%. The GSD has a narrow size distribution when GSD<2,5. l5 in some embodiments, GSD is less than 2, less than L75, or less than l5. in one embodiment, GSD is less than L8. {@153} While, the drug carriers are discussed in terms of coated particles, there are at least eight types of drug carriers that can he formulated with the active agent and one or more additional components. Different types of drug carriers 2% can he as follows: ((l) drug carriers comprising a core formed by the active agent to which the additional component absorbs/adsorbs or the additional component forms one or more coating layers on the drug carrier core; (2) drug carriers comprising a generally homogeneous mixture of the active agent and the additional component; (3) drug carriers comprising a core comprising a generally homogeneous mixture of the active agent and the additional ent, and the additional component forms one or more g layers on the drug carrier core; (4) drug carriers comprising a core formed by the onal component and the active agent forms one or more g layers on the drug r core; {5) drug rs comprising a core comprising a lly neous mixture of the active agent and the additional component, and the active agent forms one or more coating over the drug carrier core; (6) drug carrier comprising a core of material other than the active agent and the additional component, and a e of the active agent and the additional ent forms one or more coating layers on the drug carrier core; (7) drug carriers comprising a core comprising a lly homogeneous mixture of the active agent and the additional component, and a material other than the active agent or the additional component forms one or more coating layers on the drug carrier core; (3) liposomes comprising the active agent; (9) emulsions, e.g., oil/water/oil or water/oil/water emulsions; (l0) micelles; (ll) globules; (l2) suspensions; (13) dispersions; (l4) vesicles; (l5) aggregates; and (lo) drug carrier comprising any of the drug carriers of (l)«(l5) ill and further comprising one or more layers of a material other than the active agent or the additional ent. in drug carriers of (l6), the lirrther layer can he the outermost layer, a first layer on the core, interspersed between the layers described in (l)-(l5), or any combinations thereof. Without limitations, the coating layer can comprise components other than indicated above. For example, l5 the above ted g component can be mixed with other molecules or compositions to form the g layer. This can be useful in instances wherein the specified component may not be able to form a coating layer by . in some embodiments, the particle comprises a core comprising the active agent and the additional component forms one or more (e.g., l, 2, 3, 4, 5, 6, 7, 8, 9, lb or more) coating layers on the core. ltltllSéil in some embodiments, the drug canier can be in the form of a liposome. As used herein, a liposome is a structure having lipid-containing membranes enclosing an aqueous interior. Liposomes can have one or more lipid membranes. Qligolamellar, large vesicles and multilarnellar vesicles have multiple, y concentric, membrane layers. Liposomes with several nonconcentric membranes, i.e., several r vesicles contained within a larger vesicle, are termed multivesicular vesicles. ltltllSSl mes can further comprise one or more additional lipids and/or other components such as sr e.g., cholesterol. Additional lipids can be included in the liposome compositions for a variety of purposes, such as to prevent lipid oxidation, to stabilize the bilayer, to reduce aggregation during formation or to attach ligands onto the me e. Any of a number of additional lipids and/or other components can be present, ing amphipathic, neutral, cationic, anionic lipids, and programmable fusion lipids. Such lipids and/or components can he used alone or in combination. in addition to the lipids, the liposome can comprise one or more of the additives described in the disclosure. liltilSdl Liposome compositions can be prepared by a variety of methods that are known in the art. See e.g., US, Pat. Nos. 4,235,871; 4,737,323; 4,397,355 and S,l7l,678; published international Applications W0 961’l4657 and WO 96/37194; it) r, P. L. at nth, Proc. Natl. Acaa’. Sci, USA H987) 8:7413—74l7, Bangham, at of. M. Moi. Biol. 8965) 23:238, Olson, at all. Biochim. Biophys. Acta (l979) 557:9, Szoka, at at. Proc. Natl. Astral. Sci. 0:978) 75; 4l94, Mayhew, at at’. m. Biophys. Acid { 1984) 775:l69, Kim, et a2. Biochlm. Biophys. Acta @983) , and Fukunaga, et a1, Endocrine]. (£784) ll5:757. lilillfi’l’l in some embodiments, the drug carrier can he micelle. As used herein, “micelles” are a particular type of molecular assembly in which amphipathic molecules are arranged in a spherical ure such that all hydrophobic portions on the les are directed inward, leaving the hydrophilie portions in contact with the surrounding aqueous phase. The se arrangement. 2t) itfilngl in some embodiments, the drug carrier can be an emulsion. As used herein, “emulsion” is a heterogeneous system of one liquid dispersed in another in the form of droplets. ons are often biphasic systems comprising two immiscible liquid phases tely mixed and dispersed with each other. Either of the phases of the emulsion can he a semisolid or a solid, as is the case of emulsion—style ointment bases and creams. The active agent can be present as a solution in either the aqueous phase, oily phase or itself as a separate phase. liltilfi9l in some embodiments, the drug carrier can be formulated as microemulsions. As used herein, “microemulsion” refers to a system of water, oil and amphiphile which is a single optically isotropic and dynamieally stable 3Q liquid solution. Microemulsions also include thermodynamically stable, pically clear dispersions of two immiscible liquids that are stabilized by interfacial films of euactive molecules. ltltlldtll The application of emulsion formulations via dermatological, oral and parenteral routes and methods for their manufacture have been reviewed in. the U1 literature, for example see ldson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds), 1988, Marcel Bekker, lnc., New York, N.Y., volume l, p. l99; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds), l983, Marcel Dekker, inc, New York, NY” volume l, p. 245; and Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds), 1988, ll) Marcel Dekker, inc, New York, N.Y., volume l, pF 335, contents of which are herein incorporated by reference in their entirety. {eaten The drug carrier can be fabricated using methods and instruments well known in the art. For example, the drug carriers can be made using microprecipitation, encapsulation, egation, hybrid of deaggregation and encapsulation, homogenization, hybrid of deaggregation and hot homogenization, or any combinations f. in some embodiments, the process of making the particles comprises the step of selecting particles of a desired size.

Formulation autures a Kimble to BART non—Dali?“ and combination AP] ltltlltiZl The disclosure provides a composition or formulation comprising a DART. The disclosure also provides a composition or formulation comprising an acterial agent as the AH, wherein the acterial agent is not a DART molecule. in some ments, the formulation comprises two or more different APls, e.g,, two different DARTS, two different antiubacterial agents which are not BART, or a DART molecule and an anti—bacterial agent which is not a DART. ln some embodiments, the BART or the antibacterial agent is formulated as drug carrier for the APl, Without tions the formulation or the composition can be formulated for stration by any appropriate route known. in the art ing, but not limited to, topical (including buccal and sublingual) 3t) and oral or parenteral routes, including intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, nasal, and rectal administration, Exemplary modes of administration include, but are not d to, topical, injection, infusion, instillation, inhalation, or ingestion. “injection” includes, without limitation, intravenous, intramuscular, intra—arterial, intrathecal, entricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intraly‘mphnodal, transtracheal, subcutaneous, subcutieular, intra- lar, sub capsular, subarachnoid, intraspinal, intracerebral, spinal, and intracissternal injection and infusion. in some embodiments, the formulation can be in the form of an oral dosage, injectable, aerosol or inhalant. {sales} in some embodiments, the formulation can comprise two or more (e.g., two, three, four, five or more) different antiwbacterial agents as the APl. For example, the formulation can comprise two different anti-acne agents as the API, in some embodiments, the ation comprises S-chloro besifloxacin and another anti—acne agent as the APl. in one embodiment, the formulation comprises besit‘loxacin and adapalene as the APl. l5 {tidied} The formulations disclosed herein can comprise several types of cosmetically-acceptable topical vehicles including, but not limited to solutions, colloidal suspensions, sions, emulsions (microemulsions, nanoernulsions, le and nonnaqueous emulsions), hydrogels, and vesicles (liposomes, niosomes, novasomes). Components and formulation methods of suitable cosmetically—acceptable topical vehicles are well known in the art and are described, for example, in US. Pat. No. 697 and US. Pat, App. Pub, No, 2805/0l42094 and No. 2005/00tl8604, int, Fat. App, Pub. No. 2006/029818 and No. EGO/062743, content of all of which is incorporated herein by reference.

Those skilled in the art will appreciate the various methods for ing these s product forms. {@6165} in some embodiments, the formulation can be in the form of a cream, oil, lotion, serum, gel, sunscreen, nail varnish, nt, foam, spray, aerosol, powder, stick, solution, suspension, dispersion, paste, peel, and impregnated fabric (eg. a ”wipe" or tissue). Generally, the ition comprises an effective amount of the active agent. As used here, the term “effective amount” is that amount of the formulation ning the active agent necessary to achieve the WO 14666 desired improvement. in some embodiments, the ation is a topical. formulation. } in some embodiments, the fonnulation can be in a form selected from the group consisting of lotions, creams, gels, emulgel, oils, serums, powders, U1 sprays, ointments, solutions, sions, dispersions, pastes, foams, peels, films, masks, patches, sticks, rollers, cleansing liquid washes, cleansing solid bars, pastes, foams, powders, shaving creams, impregnated fabric (eg. a "wipe" or tissue}, and the like. 1961671 in some embodiments, the formulation is an antimbacterial fomiulatlon. l0 in some embodiments, the composition is an acterial ition in the form of a skin care composition. As defined herein, the term “skin care composition” refers to materials d topically to the skin that benefit, improve, or enhance the condition of the skin, or treat skin suffering from an infectious or diseased condition. Such skin care itions include bases such l5 as soap bases, cosmetic bases, medicament bases, cream bases, emollient bases, and combinations thereof, as well as other bases known in the art. {139168} Without limitations, the ation can comprise any desired amount of the API, For example, the formulation can comprise from about 0.01% to about 99% (w/w or w/V) of the Al’l. in some embodiments, the formulation can comprise from about t). 1% to about 75% (w/w or w/v), from about l% to about 56 % (WI/w or w/v), from about 1.5 % to about 46% (w/w or w/v), Aid. in some ments, the formulation can comprise from about 2.5%, 3%, 3.5%, 4%, 4. %, 5%, 7.5%, 10%, 12.5%, l5%, 17.5%, 20%, 22.5%, or 25% (w/w or w/v) of the APl.

E19169} in some ments, the formulation can comprises, in addition to the API, one or more zinc compounds. Without Wishing to be bound by a theory, zinc compounds can help to suppress sebum secretion and reduce acne inflammation. Exemplary zinc compounds include, but are not limited to, zinc acetate, zinc methionine, zinc pyrrolidone carboxylic acid, zine. sulfide, zinc gluconate, zinc picolinate, zinc sulphate, zinc citrate, etc. Without limitations, the formulation can comprise any desired amount of the zinc compound. For example, the formulation can comprise from about 0.tll% to about 99% (win; or w/v) of the zinc compound. in some embodiments, the formulation can comprise from about 01% to about 75% (w/w or w/v), from about 1% to about 50 % (w/w or w/v), from about l.5 % to about 40% (w/w or w/y), from about 2% to about % (w/w or w/v), or from about 2.5% to about 25% (w/w or w/v) of the zinc compound. in some embodiments, the formulation can comprise from about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 7.5%, l.t3%, l2.5%, 15%, l7.5%, 20%, 22.5%, or % (w/w or w/v) of the zinc compound. } in some embodiments, the formulation can further se one or l0 more excipients. Without limitations, the excipient can be selected from the group consisting of emulsifiers, preservatives, surfactants, oils, lipids, waxes, stabilizers, gy modifiers or thickening agents (gelling agent), emollients, moisturizers, conditioning agents, fragrances/perfumes, potentiating agents, preservatives, opacitiers, antioxidants, cooling agents, film g agents, l5 abrasives, exfoliating agents, colorants, pH modifiers, solvents, vehicle, penetration ers, permeation enhancers, pearlizing agents, and any combinations thereof. Amount of the excipients in the formulation can range from about 5% to 99.99% (w/w or w/v). in some embodiments, the tbrrnulation comprises one or more (ERAS ingredients. {991%} Generally, the pH of intended use of the formulation will generally range from about pit 2 to about , from about pH 3 to about pli 9, from about pl—l 4 and about pit 8, or from about pH 5.0 to about pH 7.5 or from about pH 5 to about 6.5. Suitable pit adjusting agents which can be used include one or more of organic or inorganic acids and bases ing sodium hydroxide, potassium hydroxide, ammonium hydroxide, phosphate buffers, citric acid, acetic acid, fumaric acid, hydrochloric acid, malic acid, nitric acid, phosphoric acid, propionic acid, sulfuric acid, tartaric acid, triehtyl amine, and the like. {99172} Typically, the cosmetically acceptable medium for skin care compositions comprises water and other solvents which e, but are not limited to, mineral oils and fatty alcohols. The ically—acceptable medium is from about l0% to about 99.99% by weight of the composition, preferably from about 50% to about 99% by weight of the ition, and can, in the absence of other additives, form the balance of the composition.

{MEWS} As used herein the term “cosmetically acceptable medium” refers to formulations that are used to treat skin, hair and/or nails and contain one or more LII ingredients used by those slcilled in the art to ate products used to treat skin.

The ically acceptable medium can be in any suitable form, he, a , cream, emulsion, gel, thickening lotion or powder and will typically contain water, and can contain a cosmetically acceptable solvent and/or one or more surfactants. l0 ltltll7dl The ation can se one or more conventional. onal cosmetic or dermatological additives or nts, providing that they do not interfere with the mildness, performance or aesthetic characteristics desired in the final products. The CTFA (The Cosmetic, Toiletry, and Fragrance Association; now known as the Personal Care Products Council) International Cosmetic Ingredient Dictionary and Handbook, Eleventh Edition (2096), and McCutcheonis Functional Materials, North America and internationals Editions, Mt: Publishing Co. (2007) describe a wide variety of cosmetic and ceutical ingredients commonly used in skin care compositions, which are suitable for use in the compositions of the present invention. The compositions of the present invention 2t} can contain a wide range of these additional, optional components. The total concentration of added ingredients usually is less than about 20%, preferably less than about 5%, and most preferably less than about 3% by weight of the total composition. Such components include, but are not limited to surfactants, ents, moisturizers, stabilizers, film—forming substances, fragrances, colorants, chelating agents, preservatives, antioxidants, pli ing agents, antimicrobial agents, wateruproofing agents, dry feel modifiers, vitamins, plant extracts, hydroxy acids (such as alpha—hydroxy acids and beta-hydroxy acids), and sunless tanning agents. 7Sl The formulation can comprise one or more of the following basic 3t} cosmetic raw materials, including, but not limited to hydrocarbons, esters, fatty alcohols, fatty acids, emulsifying agents, humectants, viscosity modifiers, and silicone—based materials. The formulations can contain a wide range of these basic components. The total concentration of added ingredients usmlly is less than 56%, preferably less than 20%, and most preferably less than l(l% by weight of the total formulation. Those skilled in the art will appreciate the various concentrations and combinations for employing these basic components to achieve the desired product form. ltllll76l Suitable lipids which can be used include one or more of arbons, fatty ls, fatty acids, ldes or esters of fatty acids with (31-ng alkanols. Hydrocarbons can include paraffin or petroleum jelly. Fatty ll) alcohols can. e decanol, dodecanol, tetradccanol, hexadecanol or octadecanol. Fatty acids can include (jg-€24 alltanoic acids such as hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, unsaturated fatty acids such as oleic acid and linoleic acid, Glycerides can include olive oil, castor oil, sesame oil, caprylic/capric acid triglyceride or glycerol mono-, di— and tri—estcrs with palmitic andfor stearic acid.

Esters of fatty acids can include C§~C35 ls such as beeswax, carnauba wax, cetyl palmitate, lanolin, isopropyl rnyristate, pyl te, oleic acid decyl ester, ethyl oleate and {35—ng alkanoic acid esters and the like. llllll77l Suitable hydrocarbons include, but are not limited to mineral oil, isohexadecane, squalane, hydrogenated polyisobutene, petrolaturn, paraffin, inicrocrystalline wax, and polyethylene. Suitable oils can include one or more of almond oil, t seed oil, borage oil, canola oil, coconut oil, corn oil, cotton seed oil, fish oil, jojoba bean oil, lard oil, linseed oil, boiled macadamia nut oil, mineral oil, olive oil, peanut oil, safflower oil, sesame oil, soybean oil, ne, er seed oil, tricaprylin (1,2,3 trioctanoyl glycerol), wheat germ oil and the like. The preferred quantity of oil used is in the range of about 5 to about 25% w/w, and more preferably in the range of about 5% to about 20% w/w of the composition, ltltll733 Suitable esters which can be used include, but are not limited to isopropyl palrnitate, octyl stearate, caprylic/capric triglyceride, plant waxes (Canelilla, Caranauba), vegetable oils (natural glycerides} and plant oils (Jojoba). {0617?} Suitable fatty alcohols which can be used include, but are not limited to myristyl, cetyl, stearyl, isostearyl, and hehenyl. {933.8%} Suitable emulsifying agents which can be used include, but are not limited to c (TEA/K stearate (triethanolamine/potassium te), sodium lauryl stearate, sodium cetearyl sulfate, and heeswaxfBorax), nonionic (glycerol di-stearate, PEG thyleneglycoll—lGG Stearate, Polysorbate 20, th 2 and stearechtl), and cationic (distearyldimethylammonium chloride, hehenalkonium chloride and steapyriurn chloride), polymeric (acrylates/C ill—30 alkyl acrylate crosspolymer, polyacrylamide, polyquaterniumu37, propylene glycol, l0 dicaprylatc/dicaparate and P?G~l Trideceth~6), and siliconebased als (alkyl modified dimethicone copolyols), and polyglyceryl esters, and ethoxylated di~ fatty esters. Additional suitable emulsifiers/surfactant can include one or more of ionic polysorhate surfactant, Tween® 20, Tween® 4t), Tween® 6t}, Tween® 3t), Nonylphenol Polyethylene Glycol Ethers, (alltylphenol—hydroxypolyoxyethylene), l5 Poly(oxy-l,2wethanediyl), {4=nonylphenol)-omega-hydroxy-, branched (he.

Tergitol® NP—4O Surfactant), Nonylphenol Polyethylene Glycol Ether mixtures (ie. Tergitol® NPwi’G (70% AG) Surfactant), phenoxypolyethoxyethanols and polymers thereof such as Triton®, l’oloxamer-Qi), Spans®, Tyloxapol®, different grades of Brij, sodium l sulfate and the like. The preferred quantity of the emulsifiers/surfactant used is in the range of about t).l% to about lG% w/w of the composition £66181} Exemplary htunectants for use e, but are not limited to propylene glycol, sorhitol, ne glycol, hutylene glycol, ne glycol, acetamide lVlEA (acetylethanolamlne), honey, and sodium PCA (soditunQ-pyrrolidone carhoxylate), sorhitol, triacetin, and the like. {$9182} Viscosity modifiers which can he used in the compositions of the invention include, but are not limited to xanthum gum, ium aluminum silicate, cellulose gum, and hydrogenated castor oil. {36183} Suitable thickening agents which can he used include one or more of cellulose polymer, a carbomer polymer, a cr derivative, a cellulose derivative, polyvinyl alcohol, poloxamers, polysaccharides and the like. ltlfildétl Suitable emollients which can he used include one or more of caprylic/capric triglycerides, castor oil, ceteareth-le, eth—3tl, cetearyl alcohol, ceteth 20, cetostearyl alcohol, cetyl alcohol, cetyl stearyl alcohol, cocoa butter, diisopropyl adinate, glycerin, glyceryi monooleate, glyceryl monostearate, glyceryl stearate, isopropyl ate, isopropyl palniitate, lanolin, lanolin alcohol, hydrogenated lanolin, liquid paraffins, ie acid, mineral oil, oleic acid, white petrolatum, polyethylene , polyoxyethylene glycol fatty alcohol ethers, polyoxypropylene lS—stearyl ether, propylene glycol stearate, squalane, steareth-Q or 400, stearic acid, stearyl l, urea and the like. l0 {@185} Suitable preservatives which can be used include one or more of phenoxyethanol, parahens (such as methylparaben and propylparaben), ene glycols, es, urea derivatives (such as diazolindinyl urea), and the like. liltllgol Suitable chelating agents which can he used include one or more of disodium EDTA, edetate trisodium, edetate tetrasodiurn, diethyleneamine l5 pentaacetate and the like. {39187} in some embodiments, the formulation comprises one or more of alcohols like CgcCiz alcohols, diols and triols, glycerol, methanol, ethanol, propanol, l and the like. {@3188} ln some embodiments, the formulation comprises one or more 2% tion enhancers. ary tion enhancers include anionic surfactants, such as sodium lauryl sulfate and sodium laurate; cationic surfactants, such as eetylpyridium chloride; nic surfactants, such as poloxamer, Brij, Span, Myrj, and Tween; bile salts; sodium glycodeoxycholate; sodium glycocholate, sodium taurodeoxyeholate, sodium taurocholate, Azone®g fatty acids, such as oleic and caprylic acid; cyclodextrins, such as (1—, [3—, y— cyclodextrin, methylated B— cylcodextrins; chelators, such as ESTA, sodium citrate and poly acrylates; polymers, such as chitosan, trimethyl chitosan and cationic amino acids, such as poly'-L«arginine and L~lysine Brij is the tradename for a family of nonionic polyoxyethylene commercially available from a number of suppliers. Span is the tradename for a family of sorhitan surfactants, suchas sorhitan trioleate (Span 85) and sorbitan tristearate (Span 65) and the like, commercially available from a number of ers. Myrj is a tradename for a family of polyethoxylated fatty acid commercially available from a number of suppliers, such as polyoxyethylene monostearate (Myrj 49) and the like. Tween is the tradename for a family of polyoxyethylene sorbitan or polysorhate surfactants, such as polyoxyethylene sorbitan trioleate (Tween 85) and polysorbate 8t} (Tween 80) commercially available from a number of suppliers. Alone is a tradename for l~Dodecylhexahydro—2h~Azepin~2~One ititlldgl in some embodiments, the formulation comprises one or more penetration enhancers. Exemplary penetration enhancers include, but are not l0 limited to fatty acids, bile salts, chelating agents, surfactants, and non—surfactants.

Exemplary penetration enhancers include dimethyl sulfoxide; isopropyl myristate; decyl, l or dodecyl alcohol; propylene glycol; polyethylene glycol; (39, Cit), Cll, Cl’Z or Cl2=i5 fatty ls; azone; alkyl pyrrolidones; diethoxy glycol (Transcutol); lecithin; etc. tants can also be used as penetration id enhancers. [filllglll The formulation disclosed herein can further comprise one or more optional components known for use in personal care products, provided that the optional components are physically and chemically compatible with the essential components described , or do not ise unduly impair t stability, aesthetics or performance. dual concentrations of such optional components can range from about lllltll‘i/ia to about ltl% by weight of the itions. {outfit Non—limiting examples of optional components for use in the composition e a deposition aid, ic rs, nonionic polymers, dispersed particles, conditioning agents (silicones and c conditioning oils), humectant, suspending agent, additional antindandruff actives, viscosity modifiers, dyes, nonvolatile solvents or diluents (water soluble and insoluble), pearlescent aids, additional surfactants or nonionic cosurfactants, pediculocides, pH adjusting agents, perfumes, preservatives, chelants, proteins, skin active agents, sunscreens, UV absorbers, vitamins, antioxidants, preserving agents, fillers, surfactants, UVA and/or UVB sunscreens, fragrances, viscosifying agents, wetting agents, anionic polymers, ic polymers, amphoteric polymers, viscosity/foam stabilizers, opacitying/ pearlizing agents, sequestering agents, stabilizing , humectants, anti-static agents, antifreezing agents, buffering agents, dyes, and pigments, These adjuvants are well known in the field of cosmetics and are described in many publications, for example see Harry's Book quosmericoiogy, 3th edition, Martin k)‘: Rieger, ed., Chemical Publishing, New York (2000). tEZE The compositions disclosed herein can also include a deposition aid.

The deposition aid is included to eftectively e deposition of the composition components. The deposition aid can comprise any material that enhances the deposition of the composition components onto the hair, scalp, or skin. in some embodiments, the deposition aids are cationic polymers. The concentration of the deposition aid in the ition should be sufficient to effectively enhance the tion of the components and typically range from about 0.05% to about 5%, preferably from about 0.0755945 to about 2.5%,, more preferably from about (3. l% to about l.{l%, by weight of the composition.

.... {In {$9.193} The compositions disclosed herein can comprise a cationic polymer. trations of the cationic polymer in the composition typically range from about 0.05% to about 3%, preferably from about 0.075% to about 2.0%, more preferably from about 0.le to about l.(l%, by weight of the composition.

Preferred cationic polymers will have cationic charge densities of at least about 2% 0.9 med/gm, ably at least about l..2 med/gm, more preferably at least about l5 meg/gm, but also preferably less than about 7 meg/gm, more preferably less than about 5 meg/gin. The average molecular weight of such suitable cationic polymers will generally be between about l0,000 and it) n, preferably n about Slidill) and about 5 million, more preferably between about lth,thtl and about 3 million. {9&1ng le cationic polymers for use in the compositions contain cationic nitrogen containing moieties such as quaternary ammonium or ic protonated amino moieties. The cationic protonated amines can be primary, secondary, or ry amines (preferably secondary or tertiary); depending upon the particular species and the selected pH of the composition. Any anionic counterions can be used in association with the cationic polymers so long as the polymers remain soluble in water, in the composition, or in a coacervate phase of the composition and so long as the counterions are physically and chemically compatible with the essential components of the ition or do not otherwise unduly impair product performance, stability or aesthetics. Non limiting examples of such Us counterions include halides (e.g., chloride, fluoride, bromide, iodide), sulfate and methylsulfate. Non limiting examples of cationic polymers are described in. the C’l‘PA Cosmetic ingredient Dictionary, 3rd edition, edited by Estrin, Crosley, and Haynes, (The Cosmetic, ry, and Fragrance Association, inc, Washington, DC. (1982)). l0 {@195} Non limiting examples of le ic polymers include copolymers of vinyl monomers having cationic protonated amine or quaternary ammonium onalities with water soluble spacer monomers such as acrylamide, methacrylamitle, alkyl and dialhyl acrylamides, alkyl and dialkyl methacrylamides, alltyl acrylate, alhyl rylate, vinyl caprolactone or vinyl l5 pyrrolidone, lillllllol Suitable ic protonated amino and quaternary ammonium monomers, for inclusion in the cationic polymers of the composition herein, include vinyl compounds substituted with dialkylaminoalliyl acrylate, dialkylaminoalkyl methacrylate, monoalkylaminoalkyl acrylate, monoalltylaminoallcyl methacrylate, triall<yl methacryloxyalkyl ammonium salt, trialkyl acryloxyalkyl um salt, diallyl quaternary um salts, and vinyl quaternary ammonium monomers having cyclic cationic nitrogen-containing rings such as pyridinium, olium, and quaternized pyrrolidone, e.g., alkyl vinyl imidazolium, alkyl vinyl pyridinium, alkyl vinyl pyrrolidone salts. llllllil’l’l Other suitable cationic polymers for use in the itions include copolymers of l~vinyl~2~pyrrolidone and l-vinle-methylimidazolium salt (e.g., chloride salt) (referred to in the industry by the ic, Toiletry, and Fragrance Association, “CTFA”, as Polyquaternium=l6); copolymers of l—Z— pyrrolidone and hylaminoethyl methacrylate (referred to in the industry by 3t) CTFA as Polyquaternium—l l); cationic diallyl quaternary ammonium containing rs, including, for example, dimethyldiallylanunonium chloride homopolymer, copolymers of acrylamide and dimethyldiallylammonium chloride (referred to in the industry by C'l‘liA as Polyquaternium 6 and Polyquaternium 7, respectively); amphoteric copolymers of acrylic acid including copolymers of c acid and dirnethyldiallylarnmoniurn chloride (referred to in the industry by CTFA as Polyquaternium 22), terpolymers of acrylic acid with dimethyldiallylammoniurn chloride and acrylamide (referred to in the industry by CTFA as Rolyquaternium 3‘9), and terpolymers of acrylic acid with methacrylamidopropyl trimethylarnmonium chloride and methylacrylate (referred to in the industry by CTFA as Polyquaterniuni 47), ll) {fiblgdl Other suitable cationic polymers for use in the composition include polysaccharide polymers, such as cationic cellulose derivatives and cationic starch derivatives. Preferred cationic cellulose polymers are salts of yethyl cellulose reacted with trirnethyl ammonium substituted epoxide, referred to in the industry (CTFA) as Polyquaterniuni it) and available from ol Corp.

(Edison, NJ, USA) in their r LR, JR, and KG series of polymers. Other suitable types of ic cellulose e the polymeric quaternary ammonium salts of hydroxyethyl cellulose reacted with lauryl dimethyl ammonium— substituted epoxide referred to in the industry (CTFA) as Polyquaternium 24.

These materials are available from Amerchol Corp. under the ame Polymer Lid—200. {@199} Other suitable cationic polymers include cationic guar gum and derivatives thereof, such as guar hydroxypropyltrimoniuni chloride, specific examples of which include the Jaguar series commercially ble from Rhone» Poulenc orated and the N~l~lance series commercially available from Aqualon Division of es, inc. cher suitable ic polymers include quaternary nitrogen-containing cellulose ethers, some examples of which are described in US. Pat. No, 3,962,4l8. Other suitable cationic polymers include copolymers of etherified cellulose, guar and starch, some examples of which are described in US. Pat. No. 3,958,53l. When used, the cationic polymers herein are either e in the composition or are soluble in a x coacervate phase in the composition formed by the cationic polymer and the anionic, amphoteric and/or rionic detersive surfactant component described before.

Complex coacervates of the cationic polymer can also be formed with other charged materials in the composition. lilithltll Polyalhylene glycols having a molecular weight of more than about Li] lGOO are useful . Polyethylene glycol polymers usefiil herein are PEG—2M (also known as Polyox WSR® N—lflg which is available from Union Carbide and as PEGQQGO); PEG—5M (also known as ‘Polyox WSR® N—35 and Polyox WSR® N-SG, available from Union e and as PEG-5,tl09 and Polyethylene Glycol BGOaOOG); PEG—7M (also known as Polyox WSR® N—7SG available from Union Carbide); FEES-9M (also known as Polyox WSR® N-3333 available from Union Carbide); and PEG—14 M (also known as Polyox WSR® N—BOGG ble from Union Carbide). {99291} The composition can also include dispersed particles. The can include at least 4) by weight of the dispersed particles; more preferably at least 0.05%, still more preferably at least (l.l%, even more ably at least 025%, and yet more preferably at least 0.5% by weight of the sed particles. in some embodiments? it is preferable to incorporate no more than about 20% by weight of the dispersed particles, more ably no more than about l0%, still more preferably no more than 5%, even. more preferably no more than 3%, and yet more preferably no more than 2% by weight of the dispersed particles. liltiZtlZl Conditioning agents include any material. which. is used to give a particular conditioning benefit to skin. The conditioning agents . in the compositions of the present invention typically comprise a water insoluble, water sible, nonnvolatile, liquid that forms emulsified, liquid particles or are solubilized by the surfactant micelles, in the anionic detersive surfactant component (described above). Suitable conditioning agents for use in the composition are those conditioning agents characterized generally as silicones (cg, silicone oils, cationic silicones, silicone gums, high refractive silicones, and silicone resins), organic conditioning oils (cg, hydrocarbon oils, polyolefins, and fatty esters) or combinations thereof? or those conditioning agents which otherwise form liquidl sed particles in the aqueous surfactant matrix herein. {00203 The conditioning agent of the compositions can be an insoluble silicone conditioning agent. The ne conditioning agent particles can comprise volatile ne, non—volatile silicone, or combinations thereof.

Preferred are non-volatile silicone conditioning agents. lf volatile silicones are present, they will typically be incidental to their use as a solvent or carrier for commercially available forms of non-volatile silicone material ingredients, such as silicone gums and resins. The silicone conditioning agent particles can comprise a silicone fluid conditioning agent and can also comprise other ingredients, such as a silicone resin to improve silicone fluid deposition efficiency l0 or enhance glossiness of the hair. {00204} The concentration of the ne conditioning agent typically ranges from about 0.0l% to about l0%, by weight of the composition, preferably from about 0.l% to about 8%, more ably from about 0.l% to about 5%, more preferably from about 0.2% to about 3%, Non—limiting examples of le silicone conditioning agents, and optional suspending agents for the silicone, are described in US, Reissue Pat. No. , US. Pat. No. greases, and US. Pat.

No. 5,l0ti,609. The silicone conditioning agents for use in the compositions of the present invention preferably have a viscosity, as ed at 25° C, from about 20 to about 2,000,000 centistokes {“cslt”), more ably from about l,000 to about l,800,000 csk, even more ably from about 50,000 to about l,500,000 cslc, more preferably from about 100,000 to about l,500,000 cslc l00205l The dispersed silicone conditioning agent particles typically have a volume e particle diameter ranging from about 0.0l am to about 50 am. For small particle application to hair, the volume average particle diameters typically range from about 0.01 pm to about 4i um, preferably from about 0.0l pm to about 2 pm, more ably from about 0.0l pm to about 051 am. For larger particle application to hair, the volume average particle diameters typically range from about 5 ,am to about l25 pm, preferably from about l0 rim to about 90 pm, more preferably from about 15 pm to about 70 pm, more preferably from about 20 am to about 50 am. {00206} Background material on silicones ing sections discussing silicone fluids, gums, and resins, as well as manufacture of silicones, are found in Encyclopedia of Polymer Science and Engineering, vol. 15, 2d ed, pp 204—308, John Wiley a Sons, inc, {1989). {00207} Silicone tluids include silicone oils, which are flowable ne materials having a viscosity, as measured at 25“ C, less than l,000,000 csk, preferably from about 5 ask to about 000 csk, more preferably from about l00 csl: to about 0 cslt. le silicone oils for use in the compositions of the present invention e polyalltyl siloxanes, yl siloxanes, l0 polyalhylaryl siloxanes, polyether siloxane copolymers, and mixtures f.

Other insoluble, latile silicone tluids having hair conditioning properties can also be used, } Other silicone fluids suitable for use in the compositions are the insoluble silicone gums, These gums are polyorganosiloxane materials having a l5 viscosity, as measured at 25° C, of greater than or equal to 1,000,000 csk. Silicone gums are described in US. Pat. No. 4,l52,4l6; Noll and Walter, Chemistry and Technot’ogy ofSiZicones, New York: Academic Press (@653); and in General Electric Silicone Rubber Product Data Sheets SE '30, SE 33, SE 54 and SE 76.

Specific non-limiting examples of silicone gums for use in the compositions of the present invention include polydimethylsiloxane, (polydimethylsiloxane) (methylvinylsiloxane) copolymer, polydimethylsiloxane) (diphenyl siloxane)(meth.ylvinylsiloxanc) copolymer and mixtures thereof. {00209} Other non—volatile, insoluble silicone fluid conditioning agents that are suitable for use in the compositions of the t invention are those known as “high refractive index silicones,” having a refractive index of at least about L46, preferably at least about 1.48, more preferably at least about L52, more preferably at least about l.55, The refractive index of the polysiloxane tluid will generally be less than about 1.70, lly less than about L60. in this context, polysiloxane “fluid” includes oils as well as gums, {002%} Silicone fluids suitable for use in the compositions of the present invention are disclosed in US. Pat. No 55l, US. Pat. No. 3,964,500, US.

Pat. No. 4,364,837, British Pat. No, 849,433, and Silicon Compounds, Petrarch Systems, inc, {1984}, {9021i} Silicone resins can be included in the silicone conditioning agent of the compositions of the present invention, 'l‘hese resins are highly cross-linked polymeric siloxane systems. The linking is introduced. through the incorporation of trifunctional and tetrafunctional silanes with monofunctional or difunctional, or both, silanes during cture of the silicone resin. lZi Silicone materials and silicone resins in particular, can conveniently be identitied according to a shorthand nomenclature system known to those of l0 ry skill in the art as “MDTQ” lature. Under this system, the silicone is described according to presence of various siloxane monomer units which make up the silicone, Briefly, the symbol M. denotes the nctional unit (Cl-ig)3Si005; i) denotes the difunctional unit (Cl-i3)gSiO; 'l‘ denotes the trifunctional unit (Ci-ingiQ”; and {2 denotes the quadra» or tetra—functional unit l5 Sit); Primes of the unit symbols (eg, M', l)", T, and Q") denote substituents other than methyl, and must be specifically defined for each occurrence, {@213} Preferred silicone resins for use in the compositions of the t invention include, but are not limited to lViQ, MT, MTQ, MDT and MDTQ resins, Methyl is a preferred silicone substituent. Especially preferred silicone resins are 2t} MQ resins, wherein the M363 ratio is from about 95:1.0 to about 0 and the average molecular weight of the silicone resin is from about ltltltl to about 16,009. {33214} The conditioning component of the compositions of the t invention can also comprise from about 6,05% to about 3%, by weight of the composition, ably from about (108% to about 1.5%, more preferably from about i).l% to about l%, of at least one organic conditioning oil as the conditioning agent, either alone or in combination with other conditioning agents, such as the silicones (described above), {@9215} le organic conditioning oils for use as conditioning agents in the compositions of the t invention include, but are not limited to, arbon oils having at least about it) carbon atoms, such as cyclic hydrocarbons, straight chain aliphatic hydrocarbons (saturated or unsaturated), and branched chain aliphatic hydrocarbons (saturated or unsaturated}, including polymers and mixtures thereof. Straight chain hydrocarbon oils preferably are from about C to about Cl9. Branched chain arbon oils, including hydrocarbon rs, typically will contain more than l9 carbon atoms.

L11 {llllZlol ic miting examples of these hydrocarbon oils include in oil, mineral oil, saturated and unsaturated dodecane, saturated and unsaturated tridecane, saturated and unsaturated tetradecane, ted and unsaturated pentadecane, saturated and unsaturated hexadecane, polybutene, polydecene, and mixtures thereof, Branched chain isomers of these nds, as ll) well as of higher chain length hydrocarbons, can also be used, examples of which include highly branched, saturated or unsaturated, alkanes such as the permethyl— substituted s, e,g., the permethyl-substituted isomers of hexadecane and eicosane, such as 2, 2, 4, 4, 6, 6, 8, 8-dimethylwl(l~methylundecane and 2, 2, 4, 4, 6, 6—dimethyl—S—methylnonane, available from Permethyl Corporation.

Hydrocarbon polymers such as tene and polydecene are preferred. A preferred hydrocarbon polymer is polybutene, such as the copolymer of isobutylene and butene. A cially available material of this type is L—lLl polyhutene from Amoco Chemical Corporation, {@3217} Organic. conditioning oils for use in the compositions of the present invention can also include liquid polyolefins, more preferably liquid poly—dr- oletins, more preferably hydrogenated liquid poly~a~olefins Polyolefins for use herein are prepared by polymerization of {24 to about Cid olefenic monomers, preferably from about C6 to about Cl 2. {@9218} Non—limiting examples of olefenic monomers for use in preparing the ix) [.11 polyolefin liquids herein e ethylene, propylene, l-butene, l-pentene, l- , l—octene, l-decene, l~dodecene, lwtetradecene, branched chain isomers such as 4~mefhyl~l~pentene, and mixtures thereof. Also suitable for preparing the polyolefin liquids are olefin containing refinery oclcs or effluents. Preferred hydrogenated a—olefin rs include, but are not limited to: l-hexenc to is hexadecenes, inoctene to l—tetradecene, and mixtures thereof. ltltl2l9l cher suitable organic conditioning oils for use as the conditioning agent in the compositions of the present ion include, but are not limited to, fatty esters having at least it) carbon atoms. These fatty esters include esters with hydrocarhyl chains derived from fatty acids or ls (eg, mono—esters, polyhydric alcohol esters, and die- and tri~carhoxylic acid esters), The hydrocarhyl ls of the fatty esters hereof can include or have covalently bonded thereto other compatible functionalities, such as amides and alko‘xy moieties (cg, ethoxy or ether linkages, etc). liltl22tll Specific es of preferred fatty esters include, but are not limited ll) to: isopropyl isostearate, hexyl laurate, isohexyl laurate, yl palmitate, isopropyl palmitate, dccyl oleate, yl oleate, hexadecyl stearate, decyl te, dihexyldecyl adipate, lauryl lactate, myristyl lactate, cetyl lactate, oleyl stearate, oleyl , oleyl myristate, lauryl acetate, cetyl propionate, and oleyl adipate, ltltl22ll Other fatty esters suitable for use in the compositions of the present invention are mono-vcarhoxylic acid esters of the l formula R‘COQR, wherein R‘ and R are alkyl or alltenyl radicals, and the sum of carbon atoms in R’ and R is at least l0, preferably at least 2.2. @3222} Still other fatty esters suitable for use in the compositions of the present invention are di- and tri—alkyl and alkenyl esters of carboxylic acids, such as esters of C4 to C8 dicarhoxylic acids (cg. Cl to C22 esters, preferably Clto C6, of ic acid, glutaric acid, and adipic acid). Specific non—limiting examples of di- and tri-alltyl and alltenyl esters of carhoxylic acids include isocetyl stearyol stearate, diisopropyl adipate, and tristearyl citrate. in some embodiments, the composition comprises ester of at least one of lauric acid, and succinic acid have additional anti—acne and / nflammatory properties. 539223} 0ther fatty esters suitable for use in the itions of the present invention are those known as polyhydric alcohol esters. Such polyhydric alcohol esters e alkylene glycol esters, such as ethylene glycol mono and di-fatty acid esters, diethylene glycol mono- and di-fatty acid esters, polyethylene glycol mono- and til—fatty acid esters, propylene glycol mono— and disfatty acid esters, polypropylene glycol monooleate, opylene glycol 20th monostearate, ethoxylatecl propylene glycol rnonostearate, yl mono— and diwfarty acid esters, polyglyeerol poly—fatty acid esters, ethoxylated glyceryl monostearate, l,3-— butylene glycol monostearate, l,3—b.utylene glycol distearate, polyoxyethylene polyol fatty acid ester, sorhitan fatty acid esters, and polyoxyethylene an fatty acid esters. {@224} Still other fatty esters suitable for use in the compositions of the present invention are glycerides, including, but not limited to, mono-, di—, and tri— glycerides, preferably di- and triglycerides, more preferably triglycerides. For use it) in the compositions bed herein, the glycerides are preferably the mono—, cli~, and tri-esters of glycerol and long chain carboxylic acids, such as Cit) to C22 carboxylic acids, A variety of these types of materials can be obtained from vegetable and animal fats and oils, such as castor oil, safflower oil, cottonseed oil, corn oil, olive oil, cod liver oil, almond oil, avocado oil, palm oil, sesame oil, lanolin oil and soybean oil. Synthetic oils e, but are not limited to, triolein and tristearin yl ate. {$9225} Other fatty esters suitable for use in the compositions of the present invention are water insoluble synthetic fatty esters. {88226} Specific non~limiting examples of suitable synthetic fatty esters for use 29 in the itions of the present invention include: P-43 t} triester of trimethylolpropane), MCP—684 (tetraester of 3,3ndiethanol—l,5 pentacliol), MCP 2i t} diester of adipic acid), all of which are available from Mobil Chemical Company. {@8227} Also suitable for use in the compositions herein are the conditioning agents described by the Procter & Gamble Company in US. Pat, Nos. 5,674,478, and 5,750,l22. Also suitable for use herein are those conditioning agents described in US. Pat. No. 4,529,586 (Clairol), US, Pat. No. 4,507,280 (Clairol), US. Pat. No. 158 (Clairol), US. Pat. No. 4,197,865 (L'Oreal), US. Pat. No, 4,2l7,9l4 (L'Oreal), US. Pat, No, 4,38l,9l9 (l_.'0real), and US. Pat. No. 4,422,853 (L‘Oreal). {93223} The compositions can contain a humectant. The humectants can be selected from the group consisting of polyhydric ls, water soluble alkoxylated nonionic polymers, and es thereof. The humectants, when used herein, are preferably used at levels by weight of the composition of from about (.11 43.1% to about 29%, more preferably from about 0.5% to about 5%, {9622?} Polyhydric alcohols useful herein include glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, ethoxylated glucose, l,2«hexane diol, hexanetriol, dipropylene glycol, erythritol, trehalose, erin, xylitol, maltitol, maltose, glucose, fructose, sodium chondroitin e, sodium hyaluronate, id sodium adenosine phosphate, sodium lactate, pyrrolidone carbonate, glucosamine, extrin, and mixtures thereof. illiBZEiil Water soluble alkoxylated nonionic polymers useful herein include polyethylene glycols and polypropylene glycols having a molecular weight of up to about lililt’) such as those with CTFA names PEG-200, PEG-400, PEG-600, PEG—lililil, and mixtures thereof. lilll23ll The itions of the t invention can further se a suspending agent at concentrations effective for suspending water—insoluble material in dispersed form in the itions or for modifying the viscosity of the composition. Such trations range from about il.l% to about l0%, preferably from about {13% to about 5.0%, by weight of the compositions. @9232} Suitable suspending agents include crystalline ding agents that can be categorized as acyl derivatives, long chain amine oxides, or combinations f. These suspending agents are described in US. Pat. No. 4,74 l ,855. 539233} The compositions can contain also vitamins and amino acids such as: water soluble vitamins such as vitamin Bl, BB, as, Bl2, C, pantothenic acid, pantothenyl ethyl ether, nol, biotin, and their derivatives, water soluble amino acids such as asparagine, alanine, tryptophan, glutamic acid and their salts, water insoluble vitamins such as vitamin A, D, E,Aand their derivatives, water insoluble amino acids such as tyrosine, tryptamine, and their salts. {@234} The formulations disclosed herein can also contain pigment materials such as nitroso, monoazo, diazo, carotenoid, nyl methanes, triaryl methanes, xanthenes, quinolines, oxazines, azines, anthraquinones, indigoids, thionindigoids, quinacridones, ocyianines, cals, and natural colors including water soluble dye components. The compositions of the present invention can also n chelating agents. {@235} in one embodiment, the formulation is a moisturizer cream/gel base.

For example, the formulation ses at least one moisturizing agent.

Generally, the formulation can comprise from about il.{ll% (by weight) to about 50% (by weight) of the moisturizing agents to impart a moisturizing benefit upon use. it is noted that dryness is one of prime concerns of art known cne it) topical products. Exemplary moisturizing agentsinclude, but are not limited to, N— Acetyl ethanolamine, aloe vera gel, arginine PCA, chitosan PCA, copper PCA, Corn glycerides, dimethyl imidazolidinone, fructose, glucamine, glucose, glucose ate, glucuronic acid, glutamic acid, eth—7, glycereth—l2 , glycereth—Ztl , glycereth~26, glycerin, honey, hydrogenated honey, hydrogenated starch hydrolysates, hydrolyzed corn starch, lactamide MESA, lactic acid, lactose lysine PCA, mannitol, methyl gluceth—ltl, methyl glueetth‘l, PCA, PEG-=2 lactamide, PEGle propylene glycol, polyamino acids, polysaccharides, polyarnino sugar condensate, potassium PCA, propylene glycol, propylene glycol citrate, saccharide hydrolysate, saccharide isomerate, sodium aspartate, sodium lactate, sodium PCA, sorbitol, TEA-lactate, TEA—PCA, urea, l, panthenol, petrolatum, mineral oil, lanolin, lanolin alcohol, erol, esters of tocopherol, alltyl polydimethylsiloxanes, vegetable oils, hydrogenated vegetable oils, fatty acid esters, beeswax, hydrolyzed keratin, hydroxyethyl urea, carboxylie acid amides, mucopolysaecharides, and nized nitrogen rizing agents. es of quaternary nitrogen moisturizing agents include, but are not limited to, hydroxypropyl bis-bydroxyethyldimonium chloride (available as COLATMMoist 206 from Colonial Chemicals, into), moisturizing agents described in United States Patent No. 6,869,977 (content of which is incorporated herein by reference), choline salts described in United States IPatent Nos. 6,475,965 and 6,265,364 (contents of both of which are incorporated herein by reference), camitine, and ations thereof, The moisturizing agent can be present in the iation in any desired amount to give a desired ievei of moisturization. in some embodiments, the rizing agent can he preset in an amount of (i to about 5. in another embodiment, the quaternary nitrogen moisturizing agent is present in an amount of about 0.1 to about 1% by weight, in another embodiment, S the rizing agent is present at about 1% by weight. {@6236} in some embodiments, the formuiation comprises at ieast one of giyeoiie acid, iaetie acid, suifur, saiieyiie acid, and resoreinoi.

E36237} Some exempiary formulations are described in Tahies 2-5.

Tahie 2: Some exempiary erearn formuia‘tiens.

PhaseE mingredients _— EMethed of preparation 2 A;{D 5c/e*3 ECetostearyi aleohoi ”100/3 10.0%E1) Ali ingredients of E phase A were mixed and Cetyi aieohoi heated at 70-80 C __ 10.0% ._ Steary] aieohoi 2) AH ingredients of __ 5.9% __, E E E phase B were mixed and EM”__ m V VVVVV E‘Maerogo] E stirred to get uniform F .

...., E Cetostefiagi Ether 2 E 10% 53% E soiution, then phase B Span 20 was aiso heated to 70- __ __, ‘ LEE/Qa, 869C with uous stirring Apitii_ 5.0% m m Pemuien TR E 3) Phase A was added “’“ 05% 955% into phase B with continuous stirring at EMaeregoi 73'396‘3 ECetostearyi Ether 20I 50% 5.0% B ETween 2G._ W “E 4) ingredients of phase —— 5.6% --= C was added into pre— formed cream at 49°C GE3'eeroi .0% 5,01% with continuous ng Water as to EGG as. to Hit} as. to EGG 5) Finaiiy phase i) was added to get desired pi—i i) Citric acid / NaOi—i asto pit (Eisto pH as to pH Tahie 3: Seine nry emugei formuiatiuns. 0005-2; V V V H Ingredients A C Method of nreparntiou APE 0.1% 10% j;;An ingredients of 033% Oii 50% — .... phase A was mixed and 20500; _— 3338;338:7030? ; a A tngre ien s n Steam/i aILOhOE _—, ‘ "9% A j nhase B was mixed and LOieyi aiCOhOi — 2.3% i 20% E stirred to get unifgrm Liquid Paraffin — 6.0% 6.0% soiution, then phase B Span 2;; 10% _ was aiso heated to 70' Steareth 2 __. 2.0% 2.0% 89°C: With continuous : i ~ --———~ stirring : 3) Phase A was added Steareth 20 2.0% into phase 0 with _... uous stirring at E 70:800C Pemuien _._ 35% i .

W.m:m___; 4) ients of phase Propyiene giyeoi "2.23 5.0% 00% c was added into FR? 3.Water qs. to dis. 10 (3 t0 formed nn at 40°C I 190% 100% 399% i with continuous stirrino -—0.3% 0% am 00000006230: -Citric acid6; NnOH 03.5. to-niim "(.33. to pHui added ‘10 gst {5653235 PH qs. to pH Tahie 4: Some exemniary get inrmuiatinns. 00050 ingredients A 0 C Meth APE 0,01% 0.5% 2% Dingedients ofphase ;;03 10.0% 5.0% 5.0% a MfiihfixgdwSO “ Tween 20 2.0% .... 20% 3326 “3g 1 1 1 : 3 2) A33 ingredients (if 3 3§teareth ”@399ng___________“_”_______3 phase B was mixed and 3 Carhopoi 3.0% -- 1.0% stirred to get m B i‘ Pemuien - 0.5‘” —-= 505330031; Propyiene giycoi i00% 200% 15.0% i 3) Phase A was added into phase B with Water continuous stirring . c (3.5. to 100% a . 4) ingredients of phase ii Preservative {H 0’1“ 9‘1 A’9 0 C was added into pre- formed emuision with Citric acid / NaOH continuous stirring (is. to pH ./ TEA . . 03.5. to pH ) Finaiiy phase B was added to et desired :azH Tobie 5: Some exempiary iotion formniations. these ingredients A i 3 APE arim 1.5% 3% 1) AH ingredientsof Liquid paraffin 5.0% it)6% i5.o% a ghafedA ‘22” gigs and A i ° ’ . ea e at ' mm 0” ”3% 2) Ali ingredients of Giyceryi stearate _________7' phase B was mixed and Tween 29 :O‘ifow 10.8% d to get uniform ________ Pemulen .— 0:5; __ sointion, then phase B ”“TmmT“““““m‘ ““““ was aiso heated to 70— B :ihfrjoi} 2:; 36;"0/ 86°C with continuous . /o . . /e 3.9/0 stirring nt-J gift/CO! 10.0% i0.@% i0.0% 3) Phase A wag added Water as, to . as. to into phase B with i tome i isms tome iemmmmsnmmgato C Freservative ‘ O. i % O. i ”/6 80°C i Citric acid! \1aOii, 4) ingredients of phase C i TEA was added into pre— formed iotion at 46°C D qs. to pH on. to pit with continuous stirring ) Finaiiy phase D was added to desired 38} t wishing to be bound by a , the formuiation disciosed herein can provide at ieast 1.2x increase in area under the curve in a concentration on the skin vs time piot compared with formulations known in the art. Further, the formuiation can hiii at ieast 29% more P. acnes as compared to direct application of an antibiotic. i0 {93239} The formulations disclosed herein provide formuiation teehnoiogicai advances (size optimization, surface ation, and formuiation innovations) to e specificity & efficacy by enhancing penetration d: ry to the target site (sebaceous glands); improving retention to exhibit sustained effect; or easy entry into hiofiim enveioped bacteria. innate} The disciosore further provides the use of the DARTS and formulations discioses herein for treating or preventing at ieast one bacterial infection condition in a subject. The method generaily sing administering a DART or foimniation disciosed herein to a subject in need thereof. in some embodiments, the method is for treating an acne condition in a subject. {36241} The term “acne” includes inflammatory diseases of the pilosehaceous les and/or skin glands, and commonly is characterized by papules, pustules, cysts, nodules, comedones, other blemishes or skin lesions, The term “acne” as used herein includes all known types of acne. Some types of acne which can be U1 treated with the composition of the present invention are, for example, acne vulgaris, acne comedo, papular acne, premenstrual acne, preadolescent acne, acne venenata, acne ica, pomade acne, acne detergicans, acne excoriee, gram negative acne, folliculitis barbae, folliculitis, al dermatitis, hiddradenitis ativa, cystic acne, acne atrophica, bromide acne, chlorine ill acne, acne conglobata, acne detergicans, epidemic acne, acne estiyalis, acne fulminans, halogen acne, acne indurata, iodide acne, acne keloid, acne mechanica, acne papulosa, pomade acne, premenstral acne, acne pustulosa, acne scorhutica, acne scrotulosorum, acne urticata, acne varioliformis, acne venenata, propionic acne, acne excoriee, gram negative acne, steroid acne, nodulocystic acne and acne rosacea. [titl242l Without wishing to be bound by a theory, inicronization of oxacin can have an impact on its hioactivity. For example, microniztion can enhance besiiloxacin’s bioactiyity or its retention at a desired site, Further, micronization can also effect besitloxacin’s stability and amounts in a 26 formulation. Moreover, ization can also allow optimizing ties of formulations comprising micronized besitloxacin. dE} Embodiments of the s aspects disclosed herein can also be bed by one or more of the numbered paragraphs: 1. A formulation comprising an anti—acne agent and at least one carrier or excipient, wherein the anti~acne agent is in the form of a drug carrier comprising the anti—acne agent and at least one additional compound, said additional nd selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations thereof, 2. The formulation of claim I, wherein the drug carrier is coated or uncoated. 3, The formulation of aph 1 or 2, wherein the drug carrier has a size of about 5 nrn to about 2t) pm, 4. The formulation of paragraph 1 or 3, wherein the drug carrier has a size of about 5 run to about 5 pm. 5 The ation of any of paragraphs further comprising a surface modifier on the surface of the drug r. fi. The formulation of any of paragraphs l—S, wherein the surface modifier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, oteins, cationic if} molecules, and any combinations thereof. 7. The formulation of any of paragraphs rue, wherein the carrier or excipient is selected from the group consisting of emulsifiers, preservatives, surfactants, oils, lipids, waxes, stabilizers, rheology modifiers or thickening agents (gelling agent), emollients, moisturizers, conditioning l5 agents, fragrances/perfumes, potentiating agents, preservatives, opacifiers, antioxidants, cooling agents, film forming , abrasives, exfoliating agents, colorants, pH modifiers, solvents, vehicle, penetration enhancers, pearlizing agents, and any combinations thereof, 8. The formulation of any of paragraphs l—4, n the surface of the drug 2f}! carrier is substantially free of surface modifier. 9. The formulation of any of paragraphs l—S, comprising from about fl.l% to about 50% (w/w or My) of the carrier or ent. lit. The formulation of any of aphs l—Q, wherein the formulation is formulated for topical, oral or eral administration. ll. The formulation of any of paragraphs l~lfl, n the formulation is an oral dosage, injectable, aerosol or inhalant, lotion, cream, gel, emulgel, oil, serum, , spray, ointment, solution, suspension, dispersion, paste, foam, peel, films, mask, patch, stick, roller, impregnated fabric (eg. a “wipe" or tissue), or any combination thereof. l2 The formulation of any of paragraphs l—l l, further comprising a second anti-acne agent.

The formulation of any of paragraphs l—l2, wherein the second antiuacne agent is selected from the group consisting of 8—chloro fluroquinolones, acetretin, adapalene(s), alitretinoin, alpha— or heta—hydroxy acids, antibiotics, antimicrobial peptides, antimicrobials, azelaic acid, l Ln peroxide, hexarotene, bile salts, hiofilm inhibitors, clindamycin, erythromycin, etretinate, ic acid, isotretinoin, heratolytic agents, lactic acid, lipoic acid, N—acetylcystein, natural cne , octopirox, phenoxyethanol, phenoxypropanol, pyruvic acid, resorcinol, retinoic acid, retinoid(s), lic acid, sebostats, sodium sulfacetamide, it) spironolactone, , sulfur containing 1} or L—amino acids, tazarotene, tea tree oil, tretinoin, triclosan, urea, and any combinations f. 14. The formulation of any of paragraphs l—l3, wherein the formulation comprises an 8-chloro fluoroquinolone alone or in combination with another anti-acne agent. l5 15. The formulation of any of aphs l—l4, wherein the formulation comprises besitloxacin and adapalene. l6. The formulation of any of paragraphs l~l4, wherein the formulation comprises 8—chlorofluoroquinolone and an anti-inflammatory agent. 17. The formulation of any of paragraphs l-l4, wherein the formulation comprises 8—chlorot‘luoroquinolone and retinoic acid or retinoid, 18. The formulation of any of paragraphs l—l7, wherein the second anti—acne agent is in the form of a drug carrier comprising the second anti—acne agent and at least one additional nd, said additional compound selected from the group consisting of lipids, oils, rs, peptides, proteins, ydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations f, 19. The formulation of any of paragraphs l-lS, wherein the second anti-acne agent drug carrier has a size of about 5 nm to about 50 um.

, The formulation of any of paragraphs l—l9, wherein the second anti«acne agent drug carrier has a size of about lilt) nm to about 25 um 21, The formulation of any of paragraphs i~20, whereinthe second anti—acne agent drug carrier comprises a surface modifier on the surface thereof, 22, The formuiation of any of paragraphs L421, wherein the surface modifier of the second anti~acne agent drug carrier is seiected from the group consisting of iipids, oiis, poiymers, peptides, ns, carbohydrates, giycoiipids, phosphoiipids, iipoproteins, cationic molecules, and any combinations thereof, The formuiation of any of aphs L20, wherein the surface of the second antiwacne agent drug carrier is substantiaiiy free of surface it} modifier. 24, The formulation of any of aphs i—23, r comprising an auditionai active agent.

. The formulation of any of paragraphs 1—24, wherein the additionai active agent is an anti—inflammatory-agent, ation enhancer, anti-oxidant, anti—aging agent, anti—wrinkle agent, skin whitening or bleaching agent, ultravioiet (UV) light absorbing or scattering agent, skin depigmentation agent, skin regenerative agent, scar healing agent, or any combination thereof 26. The formuiation of any of paragraphs i=25, n the additionai active agent is in the form of a drug carrier comprising a compountt seiected from the group consisting of , oiis, poiymers, es, proteins, carbohydrates, giycoiipids, phosphoiipids, iipoproteins, cationic moiecuies, and any combinations thereof. 27. The formuiation of any of paragraphs 1,-26, n the additionai active agent drug carrier has a size of about 5 nm to about iGG pm, The ation of any of paragraphs 1—27, wherein the additional active agent drug carrier has a size of about 100 nm to about 25 um. 29, The formuiation of any of paragraphs 1—28, whereinthe additional active agent drug carrier comprises a surface modifier on the surface thereof. 3G. The forntuiation of any of paragraphs i429, wherein the surface modifier of the additionai active agent drug carrier is seiccted from the group consisting of lipids, oils, rs, peptides, proteins, carbohydrates, glycolipids, phospholipids, oteins, cationic molecules, and any combinations thereof. 31. The formulation of any of paragraphs L30, wherein the e of the Us additional active agent drug carrier is substantially free of surface modifier. 32. The formulation of any of paragraphs lu3l, wherein the formulation further comprises a Zinc compound. 33. A formulation comprising an antibacterial agent and at least one carrier or iii excipient, wherein the antibacterial agent is in the form of a drug carrier comprising the cterial agent and at least one additional compound, said additional compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phosphollpids, lipcproteins, cationic les, and any combinations thereof, 34, The formulation of paragraph 33, wherein the drug carrier has a size of about 5 nm to about lot} pm.

. The formulation of aph 33or 34, wherein the drug carrier has a size ot‘ahout lOO run to about 25 pm.

The formulation of any of paragraphs 3385 further comprising a surface modifier on the surface of the drug carrier.

The formulation of any of paragraphs 3386, wherein the surface modifier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, giycolipids, phospholipids, oteins, cationic molecules, and any combinations thereof, The formulation of any of paragraphs 33—37, n the surface of the drug carrier is substantially free of surface modifier.

The ation of any of paragraphs 33~38, wherein the carrier or excipient is selected from the group ting of emulsifiers, preservatives, surfactants, oils, lipids, waxes, stabilizers, rheology modifiers or thickening agents ng agent), emollients, moisturizers, conditioning agents, hagrances/pertumes, potentiating agents, preservatives, opacifiers, antioxidants, cooling agents, film forming agents, abrasives, exfoliating agents, colorants, pH modifiers, solvents, vehicle, penetration enhancers, pearlizing agents, and any ations thereof. 4G. The formulation of any of paragraphs 33—39, comprising from about 5% to about 99% (w/w or w/y) of the carrier or excipient. 4i. The formulation of any of paragraphs 33—40, wherein the formulation is ated for l, oral or parenteral administration. ll) 42. The formulation of any of paragraphs 33—4l, wherein the formulation is an oral dosage, injectable, aerosol or inhalant, lotion, cream, gel, emulgel, oil, serum, powder, spray, ointment, solution, suspension, dispersion, paste, foam, peel, films, mash, patch, sticlr, , impregnated fabric tag. a ”wipe” or tissue), or any combination thereof. 43. The formulation of any of paragraphs 33-42. r comprising a second antibacterial agent. 44. The formulation of any of paragraphs 33—43, wherein the second antibacterial agent is in the form of a drug carrier. 45. The formulation of any of paragraphs 3344, wherein the second antibacterial agent drug carrier further comprises a compound ed from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic les, and any ations f. 46. The formulation of any of paragraphs 3345, wherein the second antibacterial agent drug carrier has a size of about 5 nm to about ltltl pm. 47. The formulation of any of paragraphs 3346, wherein the second antibacterial agent drug carrier has a size of about 1th not to about 25 pm. 48.. The formulation of any of paragraphs 33-47, whereinthe second antibacterial agent drug r comprises a surface modifier on the surface hi.) C? thereof. 49, The formulation of any of aphs 33-48, n the surface modifier of the second antibacterial agent drug carrier is selected from the group consisting of , oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations thereof. 50. The formulation of any of paragraphs 33-47, wherein the surface of the second antibacterial agent drug carrier is substantially free of surface modifier.

El. The formulation of any of paragraphs 33—50 further comprising an it) additional active agent, The fonnulation of any of paragraphs 336 l, n the additional active agent is an anti-inflammatory—agent, penetration enhancer, permeation enhancer, antioxidant, anti-aging agent, anti‘wrinlrle agent, skin whitening or bleaching agent, ultraviolet {UV} light absorbing or ring agent, skin depigmentation agent, skin regenerative agent, scar healing agent, or any combination thereof 53. The formulation of any of paragraphs 3362, wherein the additional active agent is in the form ofa drug carrier, 54. The formulation of any of paragraphs 3363, n the additional active agent drug carrier further comprises a compound selected from the group consisting of lipids, oils, polymers, peptides, ns, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations thereof.

U1 U! The formulation of any of paragraphs 33—54, wherein the additional active agent drug r has a size of about 5 nm to about tilt) pm The formulation of any of paragraphs 33—55, n the additional active agent drug carrier has a size of about 100 run to about 25 um.

The formulation of any of paragraphs 33-56, whereinthe additional active agent drug carrier comprises a surface modifier on the e thereof. 58, The formulation of any of paragraphs 33—57, n the surface modifier of the additional active agent drug carrier is selected from the group ting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations thereof.

(J: ft? The fonnulation of any of paragraphs 33-56, wherein surface of the additional active agent drug carrier is substantially free of surface modifier. 60. The ation of any of aphs 33-59, wherein the formulation further comprises a zinc compound, 6l. The formulation of any of paragraphs 33~6tl, wherein the formulation l0 comprises a moisturizing agent. 62. A Dual Action al Therapeutic {DART} molecule that has two distinct anti-bacterial. mechanisms of action. 63. A DART molecule that has a fi—lactam ring and a quinolone nucleus, or a one nucleus and a nitro-heterocycle, or a ll-lactam ring and a l5 nitroheterocycle. 64. The molecule of paragraph 62, wherein the molecule ts DNA gyrase or topoisomerase W and transpeptidase—mediated cross-linking of peptidoglycans, 65. The molecule of paragraph 62 or 63, wherein the le inhibits isoprenyl pyrophosphate and transpeptldaseunediaterl cross—linking of peptidoglycans. 66. The molecule of any of paragraphs 6264, wherein the molecule inhibits isoprenyl pyrophosphate and DNA gyrase of topoisomerase W. 67. The molecule of any of paragraphs 62-65, wherein the molecule inhibits tolate synthesis and DNA gyrase of omerase W. 68. The molecule of any of paragraphs 62—66, wherein the molecule inhibits folate synthesis and transpeptidaseumediated cross-linking of peptit‘loglycans. 69, The molecule of any of paragraphs 62~67 that inhibits DNA gyrase or topoisornerase iv and the 303 suh—unit in bacteria. 70. The moiecnle of any of paragraphs 62—68, wherein the moieenie inhibits DNA 'gyrase er topoisomerase 1V and the SSS sub-unit in bacteria.

The ie of any of paragraphs 62-69? wherein the ie inhihits transpeptidase—mediated cross~iinking of peptidogiycans and the 308 or the SOS subunit in bacteria.

The moiecnie of any of paragraphs 62—70, wherein the moieenie inhibits foiate synthesis and the 303 or the SSS snh—tmit in hacteria.

The moiecnie of any of paragraphs 62~7i wherein the meiecnie inhibits isoprenyl pyrophosphate and the 308 or the 508 subunit in bacteria. it) 74. A Baal Action Rationai Therapeutic (DART) molecule that has two distinct anti-acne mechanisms of action. 75. The moiecnie of paragraph 73, wherein the ie modniates at ieast two different targets. 76. The moiecnie cf paragraph 73 or 74 wherein the first ism is an antibacterial action and the second mechanism of action is inhibition of nocyte proiiferation and differentiation. 77. The moiecnie of any of paragraphs 73-75, wherein the first mechanism is an antibacterial acticn and the second mechanism of action is anti— inflammatory. 78. A Dnai Action Raticnai Therapeutic (DART) moiecnie which inciudes two chemicai domains, each said chemieai domain g to a distinct active site in target ceiis, wherein said chemicai domains are bound tcgether through a third domain. 79. The ie of paragraph 77, wherein the third domain is a linker. 80. The moiecnie of paragraph 77 or ‘78, n the third domain is a cieavable .

Si. The moiecuie of paragraph 77 or 78, wherein the third domain is a non- cieavabie linker.

The moiecuie of aphs 77-80, wherein said third domain is ii- hydroxynndecenic acid; i,i®~decanedici; irEvpropanedioi; 1,5-pentanedii; lil-hydroxydecenic acid; succinic; lactic acid; 3-hydroxypropionic acid; or any combination thereof.. 83. The molecule of any of paragraphs 77-8l, wherein the third domain increases an activity of at least one of the two chemical domains.

U] 84, The molecule of any of aphs 77~82, wherein the third domain has antibacterial or anti-inflammatory activity.

The molecule of any of paragraphs 6283, wherein the molecule is in the form of a drug carrier. 86, The molecule of any of paragraphs 62—84, wherein the drug carrier has a ll) size of about 5 ant to about 100 um 8’7. The molecule of any of paragraphs 62u85, wherein the drug carrier has a size of about lOG nrn to about 25 um. 88. The molecule of any of paragraphs 62-86, wherein the drug carrier r comprises a compound selected from the group consisting of lipids, oils, rs, es, proteins, carbohydrates, ipids, phospholipids, lipoproteins, cationic molecules, and any ations thereof. 89, The molecule of any of paragraphs 62—87, wherein the drug carrier further comprises an additional active agent. 90. The molecule of paragraph 88, wherein the additional active agent is an anti—inflamrnatory~agent, keratolytic agent, penetration enhancer, anti- oxidant, anti—aging agent, anti-wrinkle agent, shin whitening or bleaching agent, ultraviolet (UV) light absorbing or scattering agent, skin depigmentation agent, skin regenerative agent, scar g agent, or any combination thereof. 9t, The molecule of any of aphs 62-89, wherein surface of the drug carrier is substantially free of surface modifier, The molecule of any of paragraphs 6290, wherein the drug carrier further comprises an additional cne agent The molecule of any of paragraphs 623i, wherein the second anti—acne agent is selected from the group ting of acctretin, adapalene(s), alitretinoin, alpha— or beta-hydroxy acids, antibiotics, antimicrobial peptides, antimicrobials, azelaic acid, benzoyl peroxide, tene, bile salts, biofilm inhibitors, clindamycin, erythromycin, etretinate, giycolic acid, isotretinoin, keratolytic agents, lactic acid, lipoic acid, N— cystein, natural anti—acne agents, octopirox, phenoxyethanol, phenoxypropanol, pyruvic acid, resorcinol, retinoic acid, retinoidts), salicylic acid, sebostats, sodium sulfacetamide, spironolactone, , sulfur ning 30— or L—amino acids, tazarotene, tea tree oil, tretinoin, triclosan, urea, and any combinations f. 94, The molecule of any of paragraphs 62—92, wherein the drug carrier further ll) comprises a surface modifier on the surface thereof.

The le of any of paragraphs 62—93, n the surface modifier is a compound selected from the group consisting of lipids, oils, polymers, peptides, ns, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any ation thereof. 96, A formulation comprising a dual action rational therapeutic molecule of any of paragraphs 62-94 and at least one carrier or excipient.

The formulation of paragraph 95, wherein the carrier or excipient is selected from the group consisting of emulsifiers, preservatives, surfactants, oils, lipids, waxes, stabilizers, rheology modifiers or thickening agents (gelling agent), emollients, moisturizers, conditioning agents, fragrances/perfumes, potentiating agents, vatives, opacifiers, antioxidants, cooling agents, film forming , ves, exfoliating agents, colorants, pH modifiers, solvents, vehicle, penetration enhancers, permeation enhancers, pearlizing agents, and any combinations thereof. 98, The formulation of paragraph 95 or 96 sing from about 5% to about 99% (w/w or w/v) of the carrier or excipient. 99, The formulation. of any of paragraphs 95-97, wherein the formulation is formulated for topical, oral or parenteral administration. ltltl, ”the formulation of any of paragraphs 9598, wherein the formulation is an oral dosage, injectable, l or nt, lotion, cream, gel, emulgel, oil, serum, powder, spray, ointment, solution, suspension, dispersion, paste, foam, peel, films, mask, patch, stick, roller, impregnated fabric (eg. a ”wipe” or tissue), or any combination thereof, ltll. The formulation of any of paragraphs 535—919 further sing a second anti~acne agent, , ltlZ. The formulation of any of paragraphs ilS—ltltl, wherein the second anti— acne agent is selected from the group consisting of acetretin, adapalenet’s}, alitretinoin, alpha— or beta—hydroxy acids, antibiotics, antimicrobial es, antimicrobials, azelaic acid, benzoyl peroxide, bexarotene, bile salts, bioiilrn inhibitors, clindamycin, erythromycin, etretinate, glycolic it) acid, tinoin, keratolytic agents, lactic acid, lipoic acid, N~ cystein, natural anti—acne agents, octopirox, phenoxyethanol, ypropanol, pyruvic acid, resorcinol, retinoic acid, retinoid(sl, salicylic acid, sebostats, sodium sulfacetamide, spironolactone, sulfur, sulfur containing D— or L-amino acids, tazarotene, tea tree oil, tretinoin, triclosan, urea, and any combinations thereof. l0}, The formulation of any of paragraphs 95~lt3l, wherein the second antl~ acne agent is in the form of a drug carrier. ltl4. The formulation of any of paragraphs 95-402, wherein the second anti- acne agent drug carrier further comprises a compound selected from the group ting of , oils, rs, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combinations thereof.

The formulation of any of paragraphs 95-103, wherein the second anti- acne agent drug carrier has a size of about 5 run to about 190 um. lilo, The formulation of any of paragraphs 94—183, wherein the second anti- acne agent drug carrier has a size of about lOG nm to about 25 um. lll7. The ation of any of paragraphs , whereinthe second anti.«acne agent drug carrier comprises a surface modifier onthe e thereof. 108. The formulation of any of claims 95~l0ti, wherein the surface modifier of the second antiuacne agent drug carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycoiipids, phospholipids, lipoproteins, cationic molecules, and any ations thereof, ltlg, The formulation of any of paragraphs 95405, wherein surface of the second anti~acne agent is substantialiy free of surface modifier. 110. The , formulation of any of paragraphs 95-108 further comprising an additional active agent. 111. The formulation of any of paragraphs 95-109, wherein the additional active agent is an anti—intiammatory-agent, penetration enhancer, anti» oxidant, anti-aging agent, anti—wrinkle agent, skin whitening or bleaching agent, ultraviolet (UV) light absorbing or scattering agent, skin depigrnentation agent, shin regenerative agent, sear healing agent, or any combination thereof. 3112. The formulation of any of paragraphs 9541.0, wherein the additional active agent is in the form ot’a drug carrier. ill The ation of any of paragraphs 95-l11, wherein the additional active agent drug carrier further comprises a compound selected from the group consisting of , oils, po1yrners, peptides, proteins, ydrates, giycolipids, olipids, lipoproteins, cationic molecn1es, and any ations thereof, 114. The ation of any of paragraphs 95412, wherein the onal active agent drug carrier has a size of about 5 nrn to about 1.00 pm. 115. The formulation of any of aphs 535413, wherein the additional active agent drug carrier has a size of ahout 100 nm to about 25 pm. 116, The formulation of any of paragraphs 95—114, whereinthe additional active agent drug r ses a surface modifier on the surface thereof. 11?. The formulation of any of paragraphs 95-115, wherein the surface modifier of the additiona1 active agent drug carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic La.) 0 nioiecnles, and any combinations thereof.

WO 14666 This page is blank.

The next page is page 114 HS. The forrnuiation of any of paragraphs 93-114, wherein surface of the additionai active agent drug carrier is ntially free of surface moditier. 119. The formulation of any of paragraphs 95417, wherein the formuiation U: further comprises a zinc compound. 120, A method of treating acne condition in a subject comprising administering a therapeuticaiiy effective amount of a formulation of any of paragraphs 1— 61 and 95418, The method of any of paragraph 1E9, wherein the acne condition is caused it) by antibiotic susceptible bacterial strain, 122, The method of paragraph 119 or 120, n the acne ion is caused by antibiotic resistant bacteria.

The method of any of paragraphs 1194.21, wherein the acne condition is caused by ciindamycin-, tetracycline-, doxyeyc1ine-, or erythromycin— resistant Propionbacteriem genes, 124. The. method of any of paragraphs 119-122, wherein the acne condition is caused by ciindamycinw, tetracycline», doxycyciine—, or erythromycin- tolerant Propionbacrerium acnes, 125. A method of treating a ial infection in a subject comprising stering a therapeuticaiiy effective amount of a ation of any of paragraphs i-oi and 95-118. 126, The method of paragraph 124, wherein the infection is caused by a pathogen seiected from the group consisting of Bartoneila henseiae, Borreiia burgdonféri, Campyiabacier jejuni, obacterjhrus, Chlamydia irachomaz‘is, Chlamydia pneumoniae, Chyiamydia psirtaci. ia negevensis, ichia coli (rig, Oi57zH7 and K88), Ehrlichia chafeensz’s, Cirisl‘ridium botuiinum, Closl'ridium gens‘, Ciostridinm tetam‘, Enterococcus faecalis, Haemophilias influenzae, Haemophiiius ducreyi, Coccidioides s, Bordetelia pertussis, Cexieiia barrzetii, Ureapia.sma arealyticum, Mycopiasma genitalium, Trichomatis vaginalis, Helicobacter pylori, bacter hepaticas, Legioneh’a pneumophiia, Mycnbacterium Iabercnlosis, cterium bavis, terium africanum, Mycobacrerium ieprae, Mycabacterium asiaticum, Mycobactarium avium, i%’cabacterium cei’atum, Mycabacrerium ceionaa, Mycahactarium zum, Mycabacrerium genavense, Mycabacreriam K}! kflum, Mycobaczerium intracefluiare, Mycabacterium kansrzsii, Mycobacterium maimoense, Mycnbacterium marinum, Mycnbaclerium scrqfitiaceum, Mycabacterium simiae, Mycobacterium szuigai, Mycabacterium nicerans, Mfycnbacrerium xenapi, Carynebacrerium diprheriae, Rhadocaccus equi, tsia aeschlimannii, Rickatrsia afiicae, Rickettsia cannrii, Arcanobacterium haemaiyticum, Baciilus anihracis, Bacilius cereus, ia mnnccylogenes, Yersinia , Yersinia enterowiitica, Shigeii’a tzfizscnteriae, Neisseria meningirides, Neisseria gnnarrhoeae, Streptncoccus bovis, ncoccus hemolyticus, Streptacaccus mutrms, Srreptacnccus pyagenes, acnccus pneumnniae, iacaccus aureus, Staphyiamccus epidermidis, Staphyiacnccus niae, Staphyfococcus saprophyticus, Vibrin chaierae, Vibria parakaemaiyticus, Saimanei’ia ryphz‘, Saé’maneila parag'phi, Salmonelfa enteritidis, Treponema paiiidum, Candida, Cryptceoccus, Crypmsparidium, Giardia iambiia, Microsparidia, Plasmodium vivax, Pneumncyslis carinii, Tnxnplasma gondii, Trichophyton mentagraphytes, Enterncymzaon bieneusi, Cyciaspora cayetanensis, Encephalitozaon heliem, alitoznan cunicnii, amcng ether bacteria, archaea, protozoa, and fungi. 127. The nrethed cf paragraph i234 Or 125, wherein the infection is by an otic resistant bacteriai strain. 128. The methed cf any of paragraphs i24 or 125, n the infection is by an antibiotic susceptible hacteriai strain. 129. The methpd of any of paragraphs 124-127, wherein the formulation is administered once or daiiy to said subject as a singie dose er a piuraiity cf rinses.

Some selected definitions {96244} For convenience, certain terms ed herein, in the specification, examples and appended claims are collected herein. Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. Unless explicitly stated ise, or apparent from context, the terms and phrases below do not exclude the meaning that the term or phrase has acquired in the art to which it pertains. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims° l0 r, unless otherwise required by context, ar terms shall include ities and plural terms shall include the singular, liltiZdSl Unless defined otherwise, all technical and scientific terms used herein have the same meaning as those ly understood to one of ordinary skill in the art to which this invention pertains, Although any known methods, devices, if: and materials can be used in the practice or g of the invention, the s, devices, and materials in this regard are described herein, {@246} As used herein, the term “herein” is means the whole of the sure and as such is not meant to be limited to a particular section or subsection of the disclosure. {99247} As used herein the term “comprising” or “comprises” is used in reference to compositions, s, and respective component(s) thereof, that are essential to the invention, yet open to the inclusion of unspecified elements, whether essential or not.

} The singular terms “a,” “an,” and “the” include plural referents unless context clearly indicates otherwise. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. {£99249} Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as ed in all instances by the term “about.” The term “about” when used in connection with percentages can mean $596, 324%, i394), i2.5%, i2%, il.5%, ill/o, or itl.5% of the value being referred to. {00250} Although s and materials similar or equivalent to those described herein can be used in the practice or testing of this sure, suitable methods and materials are described below, The term “comprises” means “includes.” The abbreviation, “eg.” is derived from the Latin exempli gratia, and is used herein to indicate a non-limiting example. Thus, the abbreviation “eg.” is synonymous with the term “for example.” {00251} The terms ase”, “reduced”, “reduction”, “decrease” or “inhibit” are all used herein generally to mean a se by a statistically significant amount. However, for avoidance of doubt, “reduced”, “reduction” or “decrease” or “inhibit” means a decrease by at least l0% as compared to a reference level, for e a decrease by at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a l00% decrease (lag, absent level as compared to a reference sample), or any decrease between l0= l00% as compared to a reference levelr E00252} The terms “increased”, “increase” or “enhance” or “activate” are all used herein to generally mean an se by a statically significant amount; for the avoidance of any doubt, the terms “increased”, “increase” or “enhance” or “activate” means an increase of at least l0% as compared to a nce level, for example an se of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 10 9/9 increase or any increase between l0~l00% as compared to a reference level, or at least about a 2— fold, or at least about a , or at least about a 4—fold, or at least about a 5-fold or at least about a l0-fold se, or any increase between 2-fold and l0=fold or greater as compared to a reference level. {00253} The term “statistically significant” or “significantly” refers to statistical significance and generally means at least two standard deviation (23D) away from a reference level, The term refers to statistical evidence that there is a difference. it is d as the probability of making a decision to reject the null hypothesis when the null hypothesis is actually true. ltltlZS-tl The term "glohule" as used herein refers to spherical or quasi-spherical globes, balls or other shaped particles of a substance such as form in ‘oiphasic suspensions or emulsions: Also included in the meaning of the term le" are finely divided particles of a solid material. liltiZSSl The disclosure is further illustrated by the ing examples which should not he construed as limiting. The examples are illustrative only, and are not intended to limit? in any manner, any of the aspects described herein. The following examples do not in any way limit the invention. it) EXAMPLES {@6255} Example. 1. ing of antibiotics t P. acnes strains shows that the se is unpredictable in both elindamycin=sensitive and non- responder strains EtltBZS’i’} Examples 245 and Tahles 645 describe some exemplary DART l5 molecules, their synthesis, formulations and uses.

Example 2: Synthesis of DART molecule 9 {from Table 1A} i awild‘ r8 é ”z 3 0 H 0’3 2 of}: 1' 0 3 M31) 1.5mm: as! Wif 3 - k” ’ i "V3 “g ‘fi‘m'lf. \ 3 C .I N” ——’ l F Y 3 0 5'V g / awesomeness. e x S. wanes; o i Rust RT, 15h ‘ H 0 V {£39258} Step l. Synthesis of 2: To a solution of l (l g7 2.33mmol} in mixture of dichloromethane {tonal} and dimethylthnnamide (l ml) was added N3N~ dicyelohexylcarhodiirnide (0.627g, 3 ‘94 l mrnol) followed by N~ l-lydroxyhenzatriazole g, 2.33mniol) slowly at ice cold condition and stirred at RT for 2h to obtain turbid suspension. To this turbid on pentanediol (9.85mi, 8.]8niniol) was added followed by 4—dimethylaminopyridine (0‘284g, 2,33rnrnol). The final reaction mixture was stirred at RT for 16h. The white precipitate was filtered and extracted with ethyl acetate. The filtrate was washed with brine solution, dried over sodium sulphate and evaporated to get crude mass.

The crude product was purified by flash eoiumn chromatography g with 1% methanoi/ dichiorornethane to obtain pure compound 2 (6.9g, 89% yieid). {@9259} Step7’ Synthesis of DART, 9: To a soiution of 3 (O72g, 1.55mmoi) in mixture of dichloromethane (10ml) and dimethyiformarnide (1 mi) was added L}! ohexyicarbodiimide (0.41 5g, 2.05mmoi) foilowed by N- ybenzatriazoie (0.209g, i.55rnmoi) at RT to provide turbid suspension.

The on mixture was stirred for 3 hr at RT and compound 2 (0.79g, 1.55nimoi) was added to this turbid soiution foiiowed by addition of 4- dimethyiaminopyridine (0.18%; 1.55mmoi). The finai soiutionwas stirred at RT for i6h. The precipitate was filtered, and the tiitrate was extracted with ethyi acetate. The organic iayer was washed with brine soiution and dried over sodium sulphate to obtain the crude product. The crude product was d by flash coiumn chromatography to get the iinai product (9) in 506093 isoiated yield.

Exampie 2: Synthesis of BART moiecuie 3’? (from Tabie 1A) ' 9 ' /- “(Vii/sIi IE : 0 i i Wgfis‘a V 5 3“ 9/5 p 0:; W3?” ai g ' i 6 ittmt“' amazement: 9 H N”? H“ an, 05 $0: a ,0 PR ti, ; i t 0 ’ ‘ K002, Kama. 99:0“ . ‘~ r . 3 am “W9 ,§ {99269} Step 1. Synthesis of 4: ii—Bromouhdecanoic acid (1.33g, 5.04mmoi) was pre-mixed with the methanol {0.1mi) was added into a stirred mixture of 1 (1g, 2.5Emmoi). potassium carbonate (0.243g, 1764mmoi), and dipotassium~ hydrogenphosphate (0.175g, immoi)in N,N-dimethyiacetamide (15nd) at G~5 0C.

The reaction mixture was stirred at G 0C for 5 hr and extracted with ethyi acetate (50ml) the final soiution was washed with?.. or) aqueous sodium bicarbonate soiution(10mi) foiiowed by brine on(10mi).The organic soivent was ated to give crude mass and iinaiiy purified by flash coiurnn tography. The compound was eluted with 13% methanoi/dichioromethahe to obtain pure compound, 4 (1.2g, 82%yieid). {99251} Step—2: Synthesis of DART? 87: Te a solutien cf Ccmpcund 4 (lg, 1.72mmel) in dichlcremethane (lOmi) and lmi dimethylfermarnide was added dicyclchexylearbediimide (0.46l g, 2,23rnrnel) followed by N~ Hydroxyhenzatriazole (0°232g3 1.72mmei) at RT tc e turbid suspensien.

LII The reactien mixture was stirred fer l hr at RT. To this turbid selutien 5 (0.67g, nel) was added followed by DMAP (0.2l0g, i.72mmel) and the reaction mixture was stirred at RT fer iéh. The sien was filtered and washed with brine sciatica. The erganic layer was dried ever sodium sulphate and evaparated te obtain the crude mass. Finally the crude was purified by flash column ltl chromatography using 26% methanol/dichlcremethane as eluent te ehtain the pure cempcund, 87 with 66-65% isolated yield.

Example 3: §ynthesis cf BART meiecttie 9i} (tram Table 13) {$9262} Brains—acetic acid i—chlorcmethyi—Z—(Z-methyi“5«nitre—imidazei—l—yl)— ethyl ester (it) was synthesized according t0 the fellewing scheme.

................................................................................................................................. CzN N 8! 0H H0its, {363‘ BM men Keno; star. 3:: 0 e: 45% \"3 33—35% c: 0 N H 0 Sr Nate: Cempeund 1 in the scheme cerrespends t0 compound 9% from Table 1B {£59263} Tc a stirred solution of l—Chlcrcw3~(2—methyi~5whitro-imidazelui=yi)~ 26) e-Ola (T) (0.79g, 3.6mmol) in dichlercmethane (10ml) was added dicyclehexylcarbadiimide (DCC) (0.9g, 4.3 lmmel) ed by bremcacetic acid (0.5g, 3.6 mmol) and DMAP(0.44g, 3.6 mrnel) at RT. The reaction mixture was stirred at RT for T6 h. The itate was removed by filtration and the organic layer was evaperated te get the crude that was purified by flash column chrcmatcgraphy. The final eempeund was eluted with i—2 % methahoi/diehioromethane mixture. The compound was used for the next step without further characterization. {@264} Synthesis of 7—{4—[1~Chioromethyi=2w(Z-met‘hyl—5—nitro-imia’azoflLy!)— eEhoxycarbonyimethyij—piperazih—I-yi}-6fiuoro~1~methyl—4~oxo~4H~2~thia—8b— aza-cyclobutajrajnaphthalenen3-carboxylic acid (I): To a stirred on of 6» Fluoro-t -methy1~4-oxo~7~piperazin~ i ~yi-4H—2—thia—8h-aza~ eyeiohutaiahaphthaiehe—Zi—earhoxyiic aeid, (HE) (fiflflg, G2 mmoi) in dimethylfonnamide (Mimi) was added potassium carbonate (dddg, 0.3 rmnol) t‘oiiowed by addition of compound (H) (6.1g, 0.3 mo!) and the reaction mixture it) was stirred at RT for 3h. The reaction e was diluted with ethyiaoetate, washed two times with water and fihaiiy dried over sodium sulphate to obtain the crude mass. The etude was purified by flash eoiumri chromatography whiie eiuting with 36% ol/diehioromethane mixture to obtain the pure compound (1)., i.e., Compound hit from Table i, with 36% isolated yieid. {@265} EH—NMR (4G0 MHZ, DMSQ) doom: 2.19 (3H, d, J = 6.4 Hz, CH3), 2.57 (3H, s, CH3), 2.? (4H, m, 2 x CH2), 3 (2H, s, COCHZ), 3.32 (4H, m, 2 x CH2), 3.77—3.90t’2H, ddd, J; = 3.6 Hz, J; = 12.4 Hz, J3 = 35.2 Hzs CHZCi), 4.40- 4.56 (EH, dd, J; = 9.6 Hz.) J2 = i4 Hz CHN), 4.76 (EH, d, J: 14 Hz, CHN), 5.44 (1H, d 6.9—6.1} (1H3 , J=5.6Hz CHOCQ), q, J; 3 6 Hz, J; a 12.4 Hz, CHSN}, _. 6.4 (1H, d, J; = 6.8 Hz ; ”2.8 (iH, d, J= 14 Hz, Alf-H}, 8.04 (1H, s, AruH).

ESi—Ms (tn/Z): 509 (Md-t)“. e 4: Synthesis of BART moieeuie 91 from Tahie EB {692%} EuMethyi-S-hitrdi~0xiranyimethy1-=iHuimidazoiefiV) was synthesized according to the foiiowing scheme.

G2N N ID‘ANaOH ----------------3§- DCM. 0 “’9 - RT. 3?! ”is X2605. Acetoneéwmer, e: or: WA {A we. zen -38% ..................................................................................................................................

Note: Compound 2 in the scheme corresponds to compound §i from Tabie iii iddZd7i To a stirred on of i*Chloro—3~(2~methyi—5—rtitro—imidazoi-i-yi)~ propan—Z-aoi, (1) (6.5g, 2.27mmol) in dichioromethane (3nd) was added 2t)% 1J1 Sodium idet‘dmi) at 00C. The reaction mixture was stirred for 3h at G ”C.

After 3b the reaction mixture was extracted twice with dichioromethane, the organic iayers were combined, washed with brine and finaiiy dried over sodium sulphate to obtain pure product with 99% isolated yield. iii-NEAR (466 MHZ, CDCE3)D 5ppm: 2.517 (3H, s, CH3), 2.52 (1H, to, CH3), 2.83 (1H, m, CH2), 3.38 (iii, in, CH), 4174.23 (1H, dd, J1: 6H2, J2 = 15.2Hz CH3), 4.85—4.89 (1H, J = i4.8 CH2). {@9268} Synthesis ofd-Fiuoro— 7-{4-[2~hydroxy~3~(Bumez‘hyid-niirodmidazoi—I~ ylj‘nprcpyt’]-ipzperazinmi «32.0w! ~mcthy1—4moxc-4H—2-thia-Sbwazo- eyelet:utog’ctjnaphihoiene-3~crzrboxylic acid {2): To a stirred soiution of oro- lumethy] —4-oxo—7—piperazin— i syie-t‘iH—Z—thia-=8b=aza-cyciobuta[aioaphthaienedm carboxyiic acid, (iii) {0.2g 6.57mmoi) in acetone (iSmi) was added potassium carbonate (iii ig, moi) dissoived in water (5nd), foiiowed by addition of epoxy ornidazoieiiifldifig, 0.82mmoi). The on mixture was heated at 56 ”C for 26 hr. After completion, the reaction mixture was evaporated and extracted twice with dichloromethane. The ed organic iayer was dried over sodium suiphate and evaporated to obtain the crude mass. The crude mass was purified by column chromatography by g with iii—12% methanoifdichioromethane mixture to obtain the pure compound (2), Le, Compound 9i from. Tobie i, with —30% isolated yieid. iH—NMR (400 MHZ, DMSG) dppm: 2.12 (3H, d, J: 6.4 Hz, CH3), 2.46 (3H, s, CH3), 2.58-2.60 (2H, t, J = 5.6 Hz CH2}? ), 2.65 (4H, m, 2 x CH2), 3.2 (4H, m, 2 x CH2), 3.9—4.i (2H, m, 2 x CHEN), 4.63 (iii, d, J= 14 Hz, (SHOE), 5.15 (1H. d, J = 5.2 Hz QR), 6.39 {1H, d, J= 6.4 Hz, EHSN") 6.93 (iii, d, J= 7.2 Hz 7.79 (1H, d, J: i4 Hz, APE), 8.94 (1H, s, Aruii). ESinS , Arr-H), (in/z): 532.95(M+H).

Exemgiie 5: Synthesis 91° DART me1eetr1e 94 frem Tab1e 1B .................................................................................................................... wwwww DCC. Heat, DMAP, RT, 1811 ................................................................................................................... ......

Note; Compound 5 1n the scheme eerresponds to md 941 frem Table 13 69} Synthesis Qt” dimmer—14226111321—7—[4=(5—methyI-2—axa-[Z,3jdiaxaiu¥u yimethyi,1-piperazin-J ~y!]- 442x0-4H-2—tiria-8b«aza-cyciobutaz’ajnapht‘haieneda carbaxyiic acid 1-eh[0r0meti2yl~2~(2methyl~5~nitm—imidazai~Zadj-ethyl ester (5): '10 a stirred soiutien of 6—Fluere-1=methy1—7n[4-(5wmethy1-2uexo—[1,31diox01-4~ 11) y1methy1)—piperazin~1 ~y11-=4—exe_4Ho2uthia~8b~aza—eye10buta§a§rtaphtha1erte~3= carboxylic acid, (V) (0.5g, 1.03mrn01) in DMF 1) was added DCC (8.3g, 1.41 11111101) and HOBt (0.1463 1.033mm01) f0110wed by eri 0f 1~C1110re~3—(2- methyLS—nitro—imidazol—1—y1)—prepen—2—ol, (1) (0.285g, 1.3 11111191) and EMA? ((1.13g. 1.118 mm01) at room ature. The reactiert mixture was stirred at RT fer 16 h. The precipitate was removed by fiitration and the ergariic 1ayer was evaporated to get the crude mass. Finafly it was purified by flash ediumrt chromatography elutirig with 2—4 % methane1/dieh10remethane mixture to obtain the pure cempound, (5), 1.e., Compound 24 from Tame 1, with 611% iseiated yieid.

*H—NMR (4110 MHZ, DMSG) 8ppm: 2.03 (31-1, d, J==5.6Hz, (31—13), 2.12 (31-1, 3, CH3), 2.5 (3H, s. CH3), 2.62 {41-1, m, 2 x CH2), 3.22 (4H, m, 2 x C112), 3.954.136 (2H, m, CH2C1), 4.49—4.52 (1H, t, J = 10 Hz. GEN) 4.?7 (1H, d, J = 13.2112, CHN),, 5.63(1H, d, J 3 4.4112. CHOCQ), 6.15 (111, m, CHSN), 6.7% (11-1, (1, J: 7.2, Ar-H), 7.68 (11-1. (1, J= 14. Ar—H), 7.9 (1H, s. Ar—H).

Example 6: Synthesis ofDART le lift from Table EB S-Cieroperbenzoic acid 0 E—MethyluS-n fire—tHrimidazole N3, mum—max» W3;w Iii—‘3‘ N N02 ; Dichioromathane.RT.16h Anhy.AiCli.EE?‘-yia$e§iilet {\l/VBE E 90% ocean : 50% OH nzeog . one. RT, ten ZJI 398279} Synthesis of d«hromo—i,2«eooxyhutane (i) : A Solution of 3- Chloroperbenzoic acid (55-75% pure, 1.60 g, 9.25 mmol) in 10 ml dichloromethane was added dropwise to a stirred solution of 4—bromo~l~hutene (0.5g, 3.7 mmol) in 20 ml of dichloromethane, After addition, the mixture was stirred at 25 0C. for ltih, to precipitate rohenzoic acid. E‘inaliy the reaction it) mixture was evaporated to dryness under vacuum, dissolved in ethyl acetate, washed initiaily with 4% sodium dithionate followed by saturated sodium bicarbonate and water. y the c layer was dried over sodium sulphate, evaporated and dried under vacuo to obtain the final compound (i) with 90% isoiated yield. lH—NMR (409 MHz, CDCl}) doom: 2&0 (mt, 2H? CH2) 2.58 (5, iii. l...‘ {12 08%} 2.32 (, iii m, 0Cl-ib), 3.09 (m, 1H, CH) 3.55 (t, J: 6.4, 2H, CngBr). 7ii Syntheeis of 4—Bronio—i—{2—methyi—S—nitro—imidazoi-inyiiuhutan-Z— oi (ii): To a stirred solution of 2-methyl~5—nitro~lH—imidazoie (0.8g, 6.3 mmol) in dry ethyl acetate was added anhydrous Aluminium chloride (l.67g, 12.5 mmol) at 00C and d to stir for 15 min to dissolve 2—rnethyl—5-nitro—iH—irnidazole.

After that o-l,B-enoxybutane (i), (1.9g, 12.5rnrnoi) was added dropwise into the reaction mixture and the reaction was continued for 5h at 0 0C. The reaction mixture was siowly added into ice water and pH was adjusted to l by adding concentrated i-iCl. The organic layer was separated, washed with saturated sodium bicarbonate t‘oliowed by water. The aqueous layer obtained from the first tion adjusted to pH 74 using liquor ammonia and extracted with ethyi e. The combined organic iayer was dried over sodium saiphate and evaporated in vacuo to obtain crude compoundxude was purified by flash column chromatography whiie eiuting with 2~3% methanoL/dichioromethane mixture to U: obtain the pure compound (H) with 50% isoiated yieid. ESLMS (tn/z): 277(MY. {@3272} Synthesis of 6—Fiuera{4-{Suhydsexyad—{Z-methyia—Sunitrom imidazoi-i~yi)~hutyii-piperazin-lwyiimi~methyi—éi—oxo~4H~Z-thiawdhuaza— utaiainaphthaiene-rS—carhoxyiic acid (116): To a stined soiution of 6- Fiuoro- i ~methyi«4~oxo—7—piperazin—i —yi-4i-i —2—thia~8h~aza— it) cyeiobutaia]naphthalene}~cai'boxyiic acid, (iii) (3g, 8.5 mmoi) in DMF (36nd) was added potassium carbonate (LZOgS 8.6 mmoi) foiiowed by addition of compound (it) (2g, 7.2 mmol) and the reaction e was stirred at RT for 16h.

The reaction mixture was diluted with ethyl acetate, washed twice with water and iinaiiy dried over sodium suiphate to obtain the crude mass. The crude was purified by flash column chromatography while eiuting with L‘s-5% methanol/dichioromethane e to obtain the pure compound (lid) with 2G% isoiated yieid. iH-NMR (400 MHZ, DMSO) 5ppm: 1.61468 (2H, mu CH2), 2.12 (Si-i, d, .1 = 6 Hz, CH3), 2.45 (3H, s, CH3), 2.57 (4H, in, 2 x CH2), 3.2 (4H, in, 2 x (Si-i2), 3.8-4.i (2H, m, 2 x’ CHZN), 4.44GH, m, CHOH), 5.2 (iii, hs, CHOH), 5.4 (iii, :3, J; = Soiiz, .12 = 11.6Hz, CHSN) 6.91 (iii, d, J= 7, Ar-H), 7.78 (1H, d, J: i3.6 Hz, Ar-H), 8.04 (1H, s, Ar—ii). ESLMS (mfz): 547.93(M+H)+.

Exampie 7: Synthesis oi’BAR’i‘ moieeuie 113 from Tahio iB ...............................................................................................................................

N m A“: N02 AN ”$2 NA}. kc DEC: Lauri: acid, EMA? 0” LVN d N EAL ‘V " \ Hoar. DMF,RTi15h I 0 .1 O {@273} Synthesis of Z—Bodeeanoyioxy-3~(2—methyiwfinnitro-imidazoim propyiivpipcrazin~Lyiird—i‘inoro—i-rnethyi=4—axondfimZ—thiacgh—aza- eyciohutaiafinaphthaieneuS—carhoxyiic acid (113): To a soiution of 91 {0.5g, 0.94mmoi) in dimethyiformamide {15 m1) was added N.N- dieyciohexyiearhodiintide (0.292;, 1.41 mrnoi) foiiowed by N- hydroxybenzatriazoie (0.13g, 0.94mmoi) siowly at ice eoid condition and stirred at RT for 10min to obtain turbid suspension. To this turbid soiution iauric acid U1 , 1.5 mine!) was added foiiowed by 4—dimethy1aminopyridine (0.115g, 0.94mmoi). The finai reaction mixture was stirred at RT for 16h. The white itate was fiitered and extracted with ethyi acetate. The fiitrate was washed with water and brine soiution. dried over sodium suiphate and ated to get crude mass. The crude product was purified by flash eoiumn chromatography it) g with 2—3‘3'45; methanoi/ dichioromethane to obtain pure compound 113 with % isoiated yieid. EH—NMR (400 MHZ, CDC] Lu \_/ oppm: 0.136(3H, t, J = 6 Hz, C113). 1.05-1.38(13H, m, 12112 :1, 1.48u1.70{41~1, rn £112), 1.90—1.93QH, d, J = 11.6112. CH3).2.16—2.22 (3H. rn. CH3), 2.53 (3H. 5, C113), 2.58-2.69 (2H, m, (Ii-igN ). 2.69-2.87 (41-1, m, 2 x C112). 3.2-3.4 (41-1, m, 2 x CH2), 4.1—4.25 (2H, m. 2 x CHQN). 5.0(11—1, 5.1211011), 6.129(1H. d, J: 5.2 Hz, CHSN), 6.41 {111, d, J: 6.8 Hz Ar—H), 7,92 {111. d. J 3 14.8 Hz, Ar—i-i), 8.04 (ii-i, s, Ar—I—i). ESE—MS (hi/z): 715.2(h/i+iti) Exampie 8: Synthesis of BART moieeuie 115 from Tahie 115 Jucmrwemenwic am Z-Memyl-Smitro-‘IH-Imidazoie 0 £1 W3, -—aa> MAB, N N02 : S-bromo-t-Hexene Dichmmmafihane, RT, 15?: AMY-:Eizéfthyiaceiaie, Wm: 90% gay; 6:44 (i) (it) S as e «vs1 3 N\ or: : s i («X340 0 EEK \t’ c" ~ WK”): 0221 5“ ”\ 0H F 1'”} HCi W O . ' o K2503.DMF.R1.16h m MN .

% F {1119274} Synthesis of 2—(4—hromohuty11—oxirane (1): A Soiution of 3- Chioroperbertzoic acid % pure, 4.54 g, 18.39 mmoh in 20 m1 romethane was added se to a stirred soiution of 6-hromohexne (2g, 12.26 mmol) in 20 ml of dichloromethane. After addition, the mixture was stirred at 25 0C for 16h, to precipitate 3—chlorobenzoic acid. Finally the reaction mixture was evaporated to dryness under vacuum, dissolved in ethyl e, washed initially with 4% sodium dithionate followed by saturated sodium bicarbonate and water. Finally the organic layer was dried over sodium sulphate, evaporated and dried under vacuo to obtain the final compound (l) with 90% isolated yield. ‘n—NMR {400 MHZ, coon) 5 ppm: li-l—NMR (400 NiHZ,CDC13)5 ppm: Lilli—l .6(6H. rn. CH2) 247 (iii, d, J: 2.4, OCH) 2.75 (t, J: 4. iii. 08%), 2.91 (hs, iii. QClia) 3.41 (t, J: 6.4, 2H, CquBr) . {titlZ‘I’Si Synthesis of engromdi«(Z~methyi—S-nitro~imidazoi—i—yi)—hexan-—2-— ol (ii): To a stirred solution of Z-methyl-Sunitro-lH-imidazole (0.7g, 5.5 mmol) in dry ethyl acetate was added anhydrous aluminium chloride , l l rnmol) at 0 OC and allowed to stir for l5 min to dissolve 2—methyl—5—nitro—lH—imidazole. After that 6—hromo—l32—epoxyhexane (i), (i96g. lliOmeol) was added dropwise into the reaction mixture and the reaction was continued for 5h at 0 0C. The reaction mixture was slowly added into ice water and pii was ed to l by adding concentrated llCl. The organic layer was separated. washed with saturated sodium onate followed by water. The aqueous layer obtained from the first separation adjusted to pH 7.4 using liquid. ammonia and extracted with ethyl acetate. The combined c layer was dried over sodium sulphate and evaporated in vacuo to obtain crude compound. Crude was purified» by flash column chromatography while eluting with 26% methanol/dichloromethane mixture to obtain the pure compound (ll) with 50% isolated yieldiEISl-MS (m/z): 305.95l’lVlH-i) {titty/’6} Synthesis of ro~7-{KL{S—hydroxyv-éwa-methyl—S—nitro- imidazol—i—yi)—hexyil~piperazin~i~yl}~i~rnethyl-4~oxo—éiil—2-thia~8h~aza- cyclohutala}naphthaienew3~carhoxylic acid (115}: To a stirred solution of 6— Fluoro— l —methyl-d—oxo-7—piperazim l ~yl—4H~2—thia—8l§~aza— cyclohutalulnaphthalene-B-carboxylic acid, (iii?) (HG g, 3.l6 mmol) in dimethylformarnide (30ml) was added ium carbonate (0.43g, 3.16 mmol) followed by addition of nd (ii) (0.85g, 2.63 mmol) and the reaction mixture was d at RT for th. The reaction e was diluted with etiiyi acetate, washed twice with water and tinaily dried over sodium suiphate to obtain the crude mass. The crude was purified by flash eoiumh chromatography while eEuting with. 36% ol/diehioromethane mixture to obtain the pure LJ‘I compound (115) with 20% isoiated yield. EH—Ni‘v’iR (400 MHz, DMSO) 5 ppm: i.6l-i .68t6E-E. m, CH2), 2i (3H. d, J = 6 Hz. CH3), "2.44 {3H, s, CH3) . 2.54 (4H, m, 2 XCHZ), 3.2 (4H. m. 2 XCHz). 3.9«4.E (2H. m, 2 x CHZN), 4.38(iH, d, J = i4, CHOH). 5.2 (EH, d, J = 4.4, OH). 6.38 (EH. d. J = 5.6Hz. CHSN) 6.9 (1H, d, J = 6.8. Ar—H). 7.78 (EH. d, J 2: 14 Hz. Ar—H). 8.02 (EH. 5. Ar—H). ESLMS (ht/z): 575(M+H) Exampie 9: Synthesis of DART meieeuie EE9 from Tahie EB 4inuienesulionylch'wide ' MMOH“ch: $§N02 2 w“ \! Q Ersw DMAP. my DCM LJOTS ow, 3” °C.16h RT. 3 hr F 0 ES } Synthesis of 2~(2—MethyE—5~nitro-fiH-imidezoi-l~yi) ethyi 4— methyiheezeneseifonate {i}: To a stirred soiution of 2-(2—Methyl—S—rtitro» imidazoi—E—yi)»ethanol (4g, E686 mmol) in dichloromethane (50ml; was added triethylamirie (7.3mi). 4—toulene5uifonylchloride (6.42g. 33.72 mmoi) foiiowed by 4—dimethylaminopyridinetOQg. E.68mmoi) at O 0C. The reaction mixture was stirred at RT for 3h. After completion. reaction mixture was washed with water, % HCl. sat. NaHCO3 and water. The organic layer was dried over N32804 and evaporated to get crude mass. The crude was purified by flash eoEumri chromatography by elutirig with 2—3% methatiol/diehioromethane to obtain pure nd with 90"/o isolated yield HNMR (400 MHZ CDCEg) oppm1245 (3H, N) U1 s, AruCHi3,)2 . 53HH. s. —CH3 .437(2H J = 4.8Hz, CH2), 4.54(2H, d, J = 4. 8H2 li3,) 7. 29(2H.d.J = 8.4Hz. . 7.60(2H, d. J = 8.4Hz, ArH), 7.8E(EH s, Ari-i).

{G3278l Synthesis of ro—i—rnethyl—7={4—l2—(2=ntethyl—5-=nitrouimidazol— i—yl)-ethyll—piperazin~l—ylladv-oxo—ti’ii—Zuthiawa-aza—cyclobutalalnaphthalene- 3—carbosylic acid (119): To a stirred solution of 6—Fluoro—l—rnethyl—4~oxo—7— piperazin~l~ylJillthia—8B-aza-cyclobutalolnaphthalenednearboxylic acid, (ii) .l.6g. 14.76 mmol) in dimethylforrnamide (lOGml) was added potassium carbonate (2.93g, l4.7o mmoi) followed by addition of compound (i) (2g,a 7.2 mmol) and the reaction mixture was heated at 90 0C for l6h. The reaction mixture was diluted with ethyl acetate. washed twice with water and finally dried over sodium sulphate to obtain the crude mass. The crude was purified by flash column chromatography while eluting with 46% methanol/dichloromethane mixture to obtain the pure compound (119) with 20% isolated yield. l\\/ll{ (400 MHZ, CDClg) bppm: 2.12 (3H, d, J = 6.4 Hz, CH3), 2.52 (3H, s, CH3). 2.64 (4H, m, 2 xiii-l2), 2.72(2li. t, J: 6 ill, CH2), 3.l l-3.l8 (4H, m, 2 x CH2), 4.43-4.49 (2H, m, CHZN), 5.8-5.9 (1H, q, Ji= 6.4ilz. J2: EZSH; CHEN), 6.3 (lH, d, J= 6.8Hz, Ar— (f1 H), 7.92 (ill, 5, Ar-H), 7.95 (l H. s, Ar-li). ESl-MS (”m/z): 503 (ix/HE) Example it}: Antimicrobial susceptibility of clindamycin sensitive and resistant P. times ltltil7§l The antimicrobial susceptibility of P. acne; strains t s antibiotics was determined by micro broth dilution method as follows. lfifiZgfil s: 1". acne: (MTCC l95l and CCARM9Gl0) were cultured in Brain Heart infusion Agar {Bl-lira) at 37%: for 48 hours under anaerobic condition. For MlC test, BHI broth (lGG ul) was added into all 96 wells and lOO ul of broth containing different concentrations of othin, cefoxitime? prulifloxacinr nadifloxaein, roxitrhromycin, clindamycin and oxacin were added to first well of each row (lA to ill) and serial (double) dilution was carried out for up to it) wells (column l to column it) of 96 well plate). For bacterial inoculum, P. acnes culture turbidity was adjusted to 0.5 McFarland standard (approximately l.5 x mg ) and further diluted (100 times with sterile Bill .

Diluted P. genes suspension (lOO ul) was added to each well except sterility l wells (column l2 of 96 well plate). Inoculated plates were incubated at 379C for 72 hours under anaerobic condition. After incubation, MlC was determined by adding Alarnar blue dye Results: The MlC results on P. genes strains susceptible to elindarnyein indicated that the strain is susceptible to all the antibiotics (Figure lA). interestingly, rnycin resistant strain was also resistant to a maerolide, Roxithromycin.

(Figure EB} Example ll: Beterrnination of Minimum inhibitory (Zoneentration (Wilt?) and dose response curve for compounds 95}, ill, 94, 113, HS and lid in hath elindanryeineuseeptihle (,MTCClilSl} and elindamyeln-nonresponsive it} (CCARM P. arenas strains and laboratory S. duress strains. liltlZSll Materials: Brain heart infusion broth, P. acnes strains (MTCC l95l & CCARM 90l0), S. omens MTCC 6908, 96 wells plates Autoclaveg lncubaton Anaerobic box with anaerobic gas pack, Plate reader (595 run), Alarnar blue. {@2825 Method: P. acnes were culturedin Brain Heart on. Agar {BHIA} at 37°C for 48 hours under anaerobic condition. For MIC test, Bl—ll broth {1th ul) was added into all 96 wells and lOO ul of broth containing drug was added to first well (lA to ill) and serial (double) dilution was carried out for up to it) wells (column 1 to column 10 of 96 well plate). For bacterial inoculurn, P. genes culture turbidity was adjusted to 0.5 and standard ximately 1.5 x 108 2t} cells/ml) and further diluted (lGG times with sterile Bill broth). d P. aenes suspension (ltlll all was added to each well except sterility control wells (column ii: of 96-well plate). {$9283} The plates were incubated at 37°C for 48 h under anaerobic condition for P. acnes and 37°C for 24 h for S. aureus The plates were read under Bio—Rad plate reader at 595 nrn for optical density to generate the dose—response curves.

The MIC of the test compound was recorded by addition of Alaniar blue dye. {99284} Results: Table 6 and Figures it? and 13 showed MK: and dose— response curves of different nds in both susceptible and resistant Pt acnes strains. The results showed that compound 91 showed lower MiC value and faster bacterial killing profile for both bacterial s (clindantyein—susceptible and resistant P. genes) in comparison to all other nds. in contrast compounds 1l5 and 116, which are only marginally different from 913 exhibited more than double MlC values for both P. cones strains in comparison to MIC value obtained with compound §1. Similarly, compounds 113 and Q4 hardly showed any activity YJ’I against R acnes, which could indicate the importance and involvement of free carbonyl and carbonylic groups towards better activity of the compound. interestingly the compound hi1 was found to be not active in clindamycin~ susceptibleP. cones but showed promising activity in clindamycin “nonresponder P. acues strain as observed inthe esponse curves of Figures 1C and 1D. l0 {96235} in ce of S. aureus strain all compounds lltlr 91, 115 and 116 have r activity but compound 113 showed least activities that might be due to steric constrains ed during binding to the target protein (Table 6). itltl286l sions: nd 91 is a potent anti—acne agents and an efficacious drug against treatment of both clindarnycinususceptible. ln mycin resistant P. cones strain also, it showed the highest activity which is better than in the susceptible strain. The same is also reflected in the dose response curve (Figure it: and 1D). in both s, at a very low concentration, all R genes bacterium were killed, indicating it overcomes all mechanisms of antibiotic non~responsivenessfi ie can overcome mechanisms underlying tolerance (described in introduction) as well. as resistance, This suggests the ure of compound 91 is unique that shows higher bio—efficacy in ison to all other compoundsagainst both susceptible and resistant P. cranes strains.The wide variability in outcome with structurally—related molecules also highlight the fact that it is not possible to predict efficacy just because of structural similarities Similarly, the higher MIC value for compound 331 against S aureus in comparison to P. acner proves the specificity of compound hi towards a particular bacterial strain, and that results seen in a bacterial species is not portable to r disease-causing bacteria. {lltl28‘7l Purity and bio-activity of compounds 9%, 91, 94.5 113, 115 and 116 (from Table 1A d; 13) The purity of all above mentioned DART molecules were evaluated by HilLC and their bio—activity were ted in both P. genes susceptible and resistant strains and S. oureus susceptible strain. The results are shown in Table d.

Tania 6. EFL-C purity and MHC vaines of eomoounds for both eiindainycin susceptible and ant P. dent’s strains and a laboratory 5'. agrees straint MK) fW—fi Molecules Susceptible Resistant Susceptible P aenes (195l)3 P. genes (9019) S. aureus (6998) _3___(gig/ml)strain strain (u 3 _____adamaflj 9% l7 ()9 re ___..._._l_ l.5 3 Examole 12: DNA gyrase activity assay with nds 9%? 9i, $4, 113, 115 and .‘llé with E. coir“ BNA gyrase. ll) {cease} Materials: DNA. gyrase assay kit (Topogen inc), proteinase K, chlorofonn, isoamyl alcohol, ent test compounds, agarose gel electrophoresis system } Method: DNA super—coiling activity was assayed using BNA gyrase assay kit (Topogen incl) with relaxed pHOT‘l E. coli plasmid DNA according to manufacturer’s protocol. The standard reaction mixture (20 til) contained 35 mM Tris—illC-l (pl-l 7.5), 24 ml‘vi KCl, 4 mid MgClz, 2 lel dithiothreitoldfi mM dine, l mM ATP? 55% glycerol, ill nig/ml bovine serum n (BSA), l0 pig/ml relaxed pl-iOTl plasmid DNA and l U of DNA gyrase. The reaction mixture was incubated at 37°C for l n in presence of selected compounds/fluoroquinolone at dittereiit concentrations. The reaction was terminated by addition of HS volume of loading dye (4 ul) followed by proteinaseK (50 ) and again incubated at 379C for half an hour. 26 ul chiorofonrr/isoamyi aicohoi (24:1) was added to each tube and vortexed briefly.

Thereafter, the biue s phase was ted out and ed by 1% agarose gel eiectrophoresis. The gel was stained by ethidium bromide for half an hour and destained with water for s. The gel was then visuaiized in a trans» iliuminator and photographed. {11112911} Resuits: Compounds inhibit snpencoiiing activity of fiNA gyrase: nd 91 with the best MiC vaiue was first chosen for ing its effect on DNA gyrase activity. The ity of super—coiled band was observed at five ditierent trations (in the range from 025 pM to 5 11M) {Figure 2A} There was a decrease in superucoiied band in presence of compound 91 (nearly around 543% at 0.25 nit/i to about 2% in presence of 5 11M of compound 91) as compared to compared to the untreated control (100%) (Figure 23). 11111291} i~“t.,t,rthera ali the compounds 911, 91, 94, 113., 115 and 116 were tested at two concentrations 1 pit/i and 2.5 11M to check their effect on DNA gyrase activity.

Figure 3A showed that all the compounds were able to inhibit the super-coiiing activity of DNA gyrase However, compounds 91 and 116 are highly potent in this respect as compared to the other compounds g that minor urai difference between compound 91 and 116does not affect much in gyrase binding affinity. But higher bacterial kiiling yof compound 91 than compound 116 as observed in dose response curve (Figure 16 d; 11)) proved that compound 91 might have uniquemode of activity as compared to . 111112921 When compound $1 was compared with the known t‘luoroqninoione nadifloxacin, the former showed better etticacyof inhibiting DNA super—coding by EENA gyrase than naditioxacin at both the concentrations tested (Figure 3B): Conciusions: The resuits obtained from DNA gyrase activity assay suggest that compound 91 is the most potent in binding DNA gyrase and inhibiting its action, Thus, the mechanism of inhibition of bacteriai growth is by impeding the DNA gyrase function, thus preventing important ceiluiar Emotions and eventnaily ceiinlar death. This compound shows a better antibacterial efficacy and binding affinity in comparison to a known tiuoroquinoione, naditioxacini These results are in accordance with the results obtained from the MK: data and dose response curves.

Example 13: Mutant Prevention Concentration (MFC) of compound $1 in comparison to the other known t‘luoroqulnolones {@293} als: Brain heart infusion broth, P. genes MTCC l95l, Petri plate, Autoclave, incubator, Anaerobic box with anaerobic gas pack, {@294} Method: P. genes were culturedin Brain. Heart on Agar (BlllA) at BT’C for 48 h under anaerobic condition. 3 to 4 Petri plates containing 48 h P. acnes culture was ded in sterile Phosphate buffer saline, PBS (pi-l 7.2) and it) turbidity was ed to l y of l at 630 nm (109 cells/ml} 50 ml of this culture suspension was centrifuged at 4000 rpm for 40 min. The supernatant was discarded and pellet resuspended in 250 pl of sterile PBS. 56 ul of these cell suspensions (lllw cells) was spread over the plates containing various concentrations of antibiotics (around MlC range). Plates were incubated at 37°C l5 for 48 h and the lowest concentration of the drug that allowed no growth was taken as its MPG if thin film was observed in higher antibiotic concentration plates then thin tilm was further sub—cultured on. drug free plates The growth obtained on drug free plates were then sub—cultured on plates containing drugs (at the concentration at which the thin film was isolated). if growth was not observed 2t} in these plates at the end of the incubation. period then. the same concentration was confirmed to be the MPC for the said compound. {96295} Results: The ’lahle 7 showed the value of MPC and the ratio of MPG and MlC of compound at along with other known fluoroquinolones and clindarnycin against P. genes. The MFG/MK: ratio indicates that compound 9,1 is almost 3 times more active in preventing development of resistance against P. acnes than known anti—acne antibiotic, nadifloxacin, new generation tluoroquinolone, uliiloxacin and 2 times with respect to clindamycin. MPG/Mil: ratios are found to be close sit‘loxacin and compound 91‘ This concludes both besiiloxain and compound 91 are effective molecules for treatment and El) preventing P. acnes and ideal for preventing resistance pment against pathogen.

WO 14666 {39296} Conclusions: Unlike MlC g, which typically uses an inoculuni size of approximately ,ltl‘l - lG5 cfu/rnl, the calculation of the MPG needs a lmge inoculnms (approximately ll}9 — ltll'3 chi/nil). This high inoculum is chosen to ensure the presence of firstwstep resistant mutants within the susceptible bacterial Uh population. Moreover, WC/MIC forconipound gills-only 13.3 giseiiwaeieas a is almost 8 for nadifloxacinlhe narrower the window between MK: and MPC of an antibiotic molecule the lesser than chance of selective growth of mutants in an in viva situationflhis implies that compound ‘9}; may he more effective than other known anti-acne agentsin preventing development of ance in targeted l0 es. ltltl2§7l 'l‘ahle 7: Mutation prevention concentration {MPC} and ratio of M?(fifh€l€ of the compound :9}: and comparison with known tlnorooulnolones and lineosainide Molecules ‘ MPG/MIC Example l4: Zone of inhlhitlon (ZQE) of topical gel fermulation with componnd 91 in comparison with other marketed ations } Materials: Brain heart infusion broth, S. aureus MTCC 69983 P. genes strains (M’l‘CC l951 and CCARM 90W), 96 wells plates, Autoclave, incubator, 2t) Anaerobic box with anaerobic gas pack, Plate reader (595 inn), Alamar hlue } Method: For ZOl test, ltlll gal of bacterial suspension (0.5 McFarland standard equal) was spread on Bl—lA plates. Test samples (formulations) were dissolved in water/solvent based on the solubility. Sterile disc (6 min) were loaded with ill gal of test s (with various concentrations of the compound) and were placed on the plates containing ial culture. The plates were incubated at 370C for 43 h under anaerobic condition for P. genes and 37°C for 24 h for S, aureus, followed by their ZQl measurements. itltl3tltil Results: The ZOl (measured in cm) of the different test samples are shown for P. acnes i95l (susceptible, Table it); R genes 9010 (resistant, Table §l and S. oureus (susceptible, Table to). The formulation with compound 91 showed bacterial killing profiles against both bacterial strains indicate that nd 91 retain its activity in presence of other excipients present in the formulation. stinglti ZOl data showed fast bacterial killing with resistant P. names as compared to tible R genes specially at low drug concentration (1— Zug) supported by dose se curve as seen in Figure it: and ii} as well as DNA-gyrase assays (Figure 2A a 3A), Compound Eli also works in case of 51 names strain and ZGl results support MIC values as seen in Table 5. ititlfitlil Conclusions: Formulation with compound 9i has activity against certain graniwpositive bacterial s. in case of P. acues, both. elindamycin- l5 susceptible and elindamycin non=responderstrains, the formulation remainsactivewith compound §land preferably more efficacious in elindamycin— nonresponderi’. aenes supporting the fact drug specific bio~activity.

Tables 84%. Zone of inhibition (ZQE) of topical gel formulation with compound §i in comparison for ciindarnycin tible P. genes i95iirt), elindamyeinunonresponder P. genes sore (B) and laboratory S. nurses strain Table 3 ‘ Samples Zfil tern} P. nurses i95l ptible) Formulation A (Compound iii.) 5 Formulation E (Placebo) (logo: _._l___ Table 9 Samples ZOE (em) P. ewes gillli (elindamyein resistant) Formulation E (Placebo) Talile ill gm...“ Samples ZOE (em) Si names @938 l...._w_mwm.mm.mmu.mm Formulation A (Compound 91) '* Formulation l3 (Placebo) Example 15: Determination of anti~inilammatory potential of compound 91 in YEP-=1 Cells stimulated by P: ewes lilil3tl2} Here we selected nd :91 to test nflammatory assay as it showed lower MK: and effective gyrase binding followed by faster bacterial killing profiles. Anti-inflammatory activity of compound 91 in Tlll’r-leells ll) stimulated with P. acnes (ATCC 69??) was studied. {@3393} Method: Preparation of stimulant for inflammation: P. acnes culture sion was prepared in PBS and the cell number in the suspension was adjusted to approximately 5X ll)f5 CPU/ml by measuring the cell density using a ‘Densimat, The bacterial suspension was then heat killed at 36%“ for 30 min and l5 stored at 39°C until r use. lllil3€l4l ELISA to study inflammatory response in THP—1 cells: Cells were seeded in a 96-well format (.TZXlQ5 ’l‘lll)-l eells per well) in media ning 10% PBS. The cells were stimulated to induce inflammatory cytoldnes using 3 McFarland equivalent heat~l<illed P. genes. Cells in control wells were treated with PBS, Cine hour after induction with P. genes, test agents were added to the induced cells at appropriate concentrations to he tested (compound gl at 25 ug/ml). The plates were incubated at 379C for 24 hours, After 24 h, the plates were centrifuged to pellet the cells and the supernatants were collected. The cell LI] culture atants thus obtained were analyzed for levels of cytoltines (lL—lu, llflli, lL—6 and lL—S) hy ELlSA using nan Systems kits for individual cytokines following the manufacturer’s instructions. {@395} Results: Compound 9i exerts anti-inflammatory action through the reduction of ,lL—ti in P. series—induced lllel. cells: THP—l cells, induced l0 using heat lcilled P. genes, were treated with 25 fag/ml of compound 9i following which the levels of lL-la, ill-lit, line and lL~8 were analyzed in the culture atant. At the tested concentration, compound 9i caused a significant reduction y titl%) in P. genes—induced ill—6 levels {Figure 4A and ”l‘ahle ll). Dexainethasone, the known nflammatory agent, used as a positive control, showed nearly l00% reduction in lL—e levelslhe Tl-lP-l cells showed 90% ity when treated with 25 ug/ml of compound 9i (data not shown} Compound 91 did not have an effect on lL~8 levels in the P. genes—induced TH?— l cells (Figure 43). The results presented in Figures 5A and SB show that compound 9i had a small effect on P. (genes—induced lela and lL—lh levels (approximately 29 - 25% reduction as ed in Table ll), These results suggest that nd 91 is an effective anti—inflammatory agent in a scenario specific to P. cones—induced inflammation. llltli’étldl (ionelusions: The results obtained from the BNA gyrase activity assays (cell-free system) and the anti-inflammatory assays in THP-l cells indicate that compound 9i has a dual mode of action, One of the ism of anti— bacterial effect is mediated by targeting the ial DNA gyrase in addition to ion of DNA damage from the nitroheterocycle moiety, while its anti— inflammatory properties are evident from its action on inflammatory mediators in mammalian cells. in the context of acne: this dual mode of action may aid in reducing bacterial population as well as diminishing inflammation at the site of lesions y providing faster cure and better patient compliance Tabie if: tage reductien by ecnipennd 9i in P. acaes induced different eytekine EL —ia, 1L - Eh, EL -= ti and EL — 8 reiease in THE-=1 eeiis eensidering 109% eyteirines fermatien frern P. dares d eeiis.

{Iytekines “fii‘£§n§iii§s3ii§i§5 rig/mi) Cernpeund 9f (25 gag/mi} this” mmmfiéémst/t ants % i w" this 53.83% 27.51% §9.S4% 55.76 % ["- 1L_g 58.78% ”=65 ”/0 Example 16: Tepieai Fermuiatiens with effective BART rneieeuie, 9i ititi3ti7j Beth cream and gei formulation was made with ive QART if) compound iii where cencentratien of active is typieaiiy in the range from abeut 0.5% te about 3% by weight er ativeiy 0.5% tn 2% or mest preferably 1.0% t0 1.5%. The formulatien was maintained at pi-i typieaiiy ranges frern pit 4.6 to 8.0 or preferably at pH 4.0 to 6.5 er rnest preferabiy at pH 5.0 tr) 6.0. The cencentratien of active is sufficient in reduce, treat, or prevent skin infectiens as weii as infiernrnatien at the targeted tissues caused by P. genes, S. aureus or S. epidermis or other related anaerebie grain pesitive bacteria, tigi Based en the seiubiiity protiies of nd 91, fuiiy er partiaiiy in different solvents such as dimethyl iscserbide, diethyiene giycoi mcneethyi ether, ‘PEG 400, prepyiene glycoi. benzyi alcohol, and pH 4.0 e buffer. three different iatien strategies were adapted in Obtain impreved pharnracekineties/pharntacedynarnics prefiles, high skin penetration preperties and better drug deposition characteristics, These sheuid result in faster reduction in bacteriai pepuiation aieng with quicker ion in best immune response, such as inflammatien. Such effective fernruiation with fast enset ef action weuid iinaily aliow reduction cf dose and duration of therapy hence ensuring better patient cernpiiance. These fermulatiens weuid net be restricted tn treat infections caused by P. acnes (susceptibie and resistant } but also other skin bacteriai infectiens or skin and skin structure infections or impetige er atepie dermatitis er WO 14666 rosaeea caused by different family of gram positive anaerobic bacteria such as staphylococcus Sp, streptococcus Sp. and others (susceptible and resistant strain).

More importantly this formulation should work well against resistant bacteria and prevent any further development of resistance {9336?} Composition Example l: Topical formulation with partially suspended APl (compound 9E) in gel formulation using hydroxyethyl cellulose (EEC) as a gelling agent, at pl-l 5.0-5.3 (Table l2) Table l2 (%wM) Compound 91 Active Diethylene glycol Solvent it”) ntonoethyl ether Phase A Polyethyfigne glycol 400 Propylene glycol Humeetantw—l 5.0 gy” n”? Hydroxyethyl 1 l PhaseB cellulose modifier Purified Water Vehicle Remy/Wm H Preservative 1.0 1 l emaan pnmwmm it) Etltl3ltll Method of l3reparation: l. xylethyl cellulose was added in portions into measured volume of water by maintaining stirring speed at 0 rpm The mixture was allowed to swell for 1 hour at 80400 rpm (Phase B) 2. in a te vessel, polyethylene glycol 4G0, propylene glycol, and diethylene glycol monoethyl ether were mixed together and nd $1 was added into the mixture in portions at 406 rpm for ~40-45 min to get a uniform dispersion, (Phase A) 3. The drug dispersion (Phase A.) was transferred slowly into phase B and allowed to stir at ~5G—ltltl rpm for ~30 min to form homogenous mixture. 4. Finally, benzyl alcohol was added into the final e and mixed for further 30 minutes at SG-lilll rpm to obtain a white—to—slightly yellow gel fonnulation. . y pll of the gel was maintained at 5.9 to 5.5 using citric acid solution: lilti3lll Composition e 2: Topical formulation with partially suspended APl und 91) in gel formulation using oarbopol 930 as a gelling ltl agent atpl-l 5.6-5.5. (Table 13) Table 13 ingredients Composition Function (“xii w/w) Compound in Active _[______ ll). F Diethylene glycol l monoethyl other Phase A Polyethlene llnmeotant 5 all glycol 40% Propylene glycol l-lumectant 5,6 .___w__i_____.______ Carbopol 98G l 9.6 l modifier Phase E Lm f ‘ Purified Water Vehicle as to lfill Triethanolamine loll modifier ‘ Benzyl alcohol Preservative {@9312} Method of Preparation: l5 l. Gelling agent, oarbopol 980 was added in portions to a measured volume of water ng at lGO~l 50 rpm. 2. pH of the gel mixture was adjusted to 5.5 by adding trietlianolamine solution to allow swelling of earbopol 980 in w'ater.(?hase B) 3, In a separate vessel, peiyethylene giyeoi, ene giyeei and diethyiene giyeoi menoethyi ether were mixed er and compound 91 was added into the mixture in ns whiie stirring at 400 rpm for ~ 40:45 min to get uniform dispersion. (Phase A) 4, This drug dispersion (Phase A) was added slowiy into phase B at 50-190 rpm stirring speed for about 30 min.

. Finaiiy, henzyi aieoho] was added into the finai mixture and allowed to stir for further 30 minutes at 59-10% rpm to Obtain a white-to-siightly yeiiow gei fortnuiation. i0 (3. After preparation of gel, pH was measured and fined pH was maintained at .0—5.5.

{MESS} Cnmpesitien Exanipie 3: Topieai ferniuiation with partiaiiy suspended APE (compound SEE) in gei formulation using prepyl gaiiate as idant and EDTA as a buffering agene’eheiating agent atpi-i 5.0-5.5. (Tahie Tahie i4 Cnntpositin 1 ingredients Function n {% wlw) Compound iii Active Diethyienegiyceimonflethyi * Solvent 10,0 ether : a Poiyethiene giycei 490 tant 5‘0 Propylene giycoi Humeetant 5‘0 . _ Rheeiogy Phase B WEE—iidroxyethéyi ceiluipse 1.7 : modifier “mi d Water Vehieie (is to 100 3 Ethyienediaminetetraaeetie Chieiatmg: E 3 acid dehydrate agentlbuffering (lit) Phase (3 a ent ________mm Benzyi aieohoi Preservative it) ___w _____ Prepyi gaiiate _Anti-oxidant .

Phase 3’ Citric soiutien pH medifier qs {@3714} Mettied ef Freearatien: 1. Hydrexyethy1 ee11u10se was added in portions into measured veiume 0f water by maintaining stirring speed at 190-150 rpm and aliewed to sweii fer 1 hour at 811-100 rpm. (Phase B) Ex) in a separate vessei, poiyethyiene g1yc01 400, prepyiene giyeei, and diethyiene g1yee1 meneethyi ether were mixed er and compound 91 was added into the mixture in portions whiie stirring at 400 rpm for ~40- 45 min to get uniform sien. (Phase A) . The drug dispersion (Phase A) was added into Phase B and aiiewed to stir it) 26160-160 rpm fer about 311 minutes.

Finaiiy ethyienediaminetetraacetie acid ate, benzyt aleohoi and propyi gaiiate were added te the final mixture and stirred for 3% min at 50- 1131) rpm to obtain white~to~siig1it1y yeiiew gel fonnidatient The prepared gel was maintained at p11 5.9 to 55 using citric acid soiutien. {1111315} Cempesitien Exampk 4: i fonnuiatien with fui1y suspended APi (eempeund 9i) in get formuiatien using hydrexyethyi celiuiose (BBC) as a geiiing agent at pH 5. (Terrie ES) Tabie 15: ingredients . itien I . anetiert s g {% wlw) 1 _1, Compeund 91 Active 1.0 Giyeerei Humeetent 1.0.9 E’nase A i-iydmxyetnyi Rhee1egy . 1,," ?haseB ceiiuiose modifier . _ __; ‘ Vehieie qs t0 109 Citric acid ;__pH medifier (its WO 14666 E00310} Method of Preparation: 1. Hydroxyethyi eeiiuiose was added in portions into measured voiurne of water by maintaining stirring speed at 0 rpm. The mixture was aiiowed to sweil for i it at 80—100 rpm. (Phase B) U: N in a te vessel, aqueous solution of giyoeroi was made and compound 01 was added into the mixture in portions at 400 rpm for ~40—45 min to get a uniform dispersion. (Phase A) 1.3.} . The drug dispersion (Phase A) was transferred siowiy into phase B and aiiewed to stir at ~50—i00 rpm for ~30 min to form homogenous e. 4. Finaiiy, propyi paraben was added into the finai mixture and mixed for further 30 min at 50400 rpm to obtain a white—to~sii.ghtiy yellow gel fonnuiation. 5J1 Finaiiy pH of the ,gei was maintained at 5.0 to 5.5 using citric acid solution. {00317} {Zomposition Example 5: Topieai fonnuiation with partiaiiy ded APE (compound 91) in cream formulation at pH 5.06.5 (Tania iii) Tania 16 . Compositt ients Function on (‘54: : anaum ________WW Compound 91 Active“ 1.00 E E .1 . Emoiiient and Cyciopentasaioxane 3 .00 E humeetants Cetostearyi aieohot Emoiiient 2.50 : E Phase A PEGQ Stearyi ether Emuisifier 2.00 a i E E i i E PEG-21 Stearyi ether‘E Emulsifier 2.00 Hydroxyethyi ceiiuiose _________ acetic acid dihydrate alcohol Preservative Hit} i . Bienzyl Phase Citric acid (20%w/w i H modifier s l D solution in water) : p q‘ {5353318} Method of i’reparation: l. liydroxyethy] ose was added in portions into measured volume of water by maintaining stirring speed at 500 rpm and heated at SG—SS 0C.

(Phase B} Ex) in a separate vessel, PEG—2 l ether, PEG—21 Stearyl ether and cetostearyl alcohol were heated at 50—55 9C. Cyclopentasiloxane and dimethylisosorhide were added into the mixture while stirring at 460 rpm.

Compound 91 was added into the final mixture in portions at 400 rpm for it) ~5-lt'l min to get uniform dispersion at 50 0(3.. (Phase A) (49-) . The drug dispersion (Phase A) was added slowly into Phase B and allowed to stir at ~EGQ—4QO rpm for about 20—30 min till temperature reach at 40 4. y ethylenediaminetetraacetic acid dihydrate and benzyl alcohol were 115 added to the final mixture and stirred for ~30 min at 490 rpm to obtain to-slightly yellow cream formulation. . y pH of the cream formulation is adjusted to 5.6 to 5.5 using citric acid solution. toasts} Composition Example a: "topical formulation with lly suspended APl (compound 91) in cream formulation, at pH 5,025.5 (Table l7) Table 17 Composition l ingredients Function (”/0 w/w) Phase Compound 91 ent and .

. , Cyclopentasrlexane 3 .00 l humeetant l Cetostearyl alcohel Emollient 2.50 Pee—20 Sorbitan , l m Emulsrfier i 2.60 monolauraie Sorhitan monelaurate Emulsifier 2.60 Dimethyiiseserhide Solubilizer 5m _l - WWW : Rheology i Carbopel 980 6.6 E a i’hase medifier _.,__ B Purified Water Vehicle q.s to 100 Triethanolamine i l [ Phase_______.________.___.____.__________....._...._...._.~ae.........................‘.......

‘ Benzyl alcohol Presewative {968%} Methad ef hreparatien: l. Gelling agent, earbepol 980 was added in pertiens to a measured velume of water stirring at l00»l 59 rpm. pH 0f the gel mixture was adjusted te 5‘5 with triethanelamine solution te allew swelling 0f carbopol 980 in water and heated 50-55 GC.(PhfiS€ B) u.) in a te vessel? PEG—20 an monelaurate, an monelaurate, cetestearyl alcohol, Cyclopentasilexane and dimethylisoserbide were added and heated at 50—55 0C by maintaining ng at 400—500 rpm. To l6 this final mixture compound 91 was added in pertions at 496 rpm fond—N) min to get uniform dispersien at 50 “(3. (Phase A) The drug dispersion (Phase A) was slewly transferred into Phase B at 50- 55 0C by maintaining stirring Speed 400 rpm and was cooled to 40 °C within 20—39 min. l5 Us y, henzyl aleehol was added to the final mixture and allowed to cool to room temperature to finally obtain a whitextowslightly yellow cream formulation.

Example l7: Determination of Minimum inhibitory Concentration (it/iii?) of different compounds and their formulations against P. cares strains by using micro—broth dilution method {6&3le Materials: Brain heart infusion broth? P. genes strains (MTCC 8a; Ln CCARM), 96 wells plate, Autoclave, incubator, Anaerobic box with. anaerobic gas pack, Plate reader (600 um), Alamar blue. @9322} Method: P. acnes (’MTCC 3297, MTCC l951& CCARM 9Glil) are cultured in Brain Heart infusion (Blil) Broth at 370C for 48—72 h under anaerobic condition. The test compounds / formulations are initially diluted with suitable it) solvent and further diluted with Bill broth to get the required trations.

Samples (tilt) ul) of ent concentrations (prepared by serial dilution) are added to 96-well plate. To the wells, lGO pl of P. acnes Bl-il broth culture is added [culture turbidity adjusted against 0.5 McFarland standard (approx 1.5xl08), and further diluted old with sterile Bl—ii broth], in addition. Growth l and Sterility Control are created using ltlb ul each of P. defies Bill broth culture and plain iii-ii broth, tively. {96323} Plates are incubated at 370C for 48-72 hrs under anaerobic condition.

The plate is read under d plate reader @ 595 nm for optical y to generate the dose—response curves. The MIC of the test compound is recorded by addition of Alamar blue dye. {$3324} Examples l3=22 and Tables i8-223 describe some ary novel formulations comprising stand—alone APT (e.g,, besitioxacln), either alone or in combination with adapalene.

Example i8: Micronized Besifloxacin Particle Eispersions (ill) {36325} Preparation: Besifloxacin is dispersed in surfactant solution (2% s solution of poloxamer 407). The resulting sion is passed through high re homogenizer at about Sfifibar. The output dispersion is collected in a beaker and recycled 10 times to yield a dispersion of riately sized particles (particle size range of 2pm to 8pm). The size distribution is determined by MasterSizer (Malvern instruments) and mean particle size found to be til pm [By till)—G.8urn, D‘v‘ (90)-8.9urn].

Exampte 19: Preparation et‘ Get andlCream Fermnintiens Leaded with Besifiexeein ntene, and its eemhinatien with adanatene {$3326} Ge}. and/Cream formulations containing hesifloxaein are fermutated as per the eempesitions shewn in Tnhte 18° These gei formuiations have offnwhite to stightty yellow appearance with the pH of 5-55 and ity of around 500% m‘Pas. The fermniatiens censist 0f hesifloxaein equivaient t0 1% wiw, in three different forms (1) mierenized suspended hesiflexaein HCE (Tahte 183 GL1, GL2), (2) fnlty sohibhsed hesiflexaein HCE (Tahfie 18, GL4) and (3) hesifloxaein particles suspended in cream ferrnulatien t sizing (Ch/11). in en to E6 stand atone hesiflexaein formulation, xaein is eemhined with the adapaiene (9.1%) (Tahte 18, GL1) to provide both the anti—acne and keratelytie activity in patients suffering from acne, Tahie 18: Get andKSream Fermninttnns fer Cnmnesitiens GL1, GLZ, GL3, GL4 and SME -Carbe 03940 -V Cayenne NF VV Hydrexy Pmpyl Cehuiese-H Besifioxaein HCI (equivaient to E (Di) ‘5 (D1) xaein) 5 Sod Lanryl Sulphate Light Liquid Paraffin 27} Method of preparation: (l) Allantoin is heated to 50°C to ve completely and cooled down to RT. (2) Carhopoi is added to above mixture and allowed to swell for i to 2h: (3) Dispersion of mieronized Besifloxacin and adapaiene powder is added to the swelled earhopol mixture and allowed to stir for 30 rnin at Lititlrpin. (4) Then, glyceroig propylene , PEG 4%, mer 497 is added followed by the addition of disodium EDTA solubilized in water and then add phenoxyethenol to the above stirring mixture After addition of all the l0 ingredients the mixture is allowed to stir for 3% min. (5) Above mixture is neutralized with triethanolarnine and allowed is stirring for 2—3h at 800 rpm.

Example 2%: Preparation of Cream Formniations (Cit/ii) Loaded with Besii‘loxeein HCE l5 {@323} Cream formulation ning suspended Besitloxaein particles are formulated as per the compositions shown in Tobie 18, This gel formulation has off-white to siightiy yellow appearance with the pH of 5-5.5 and viscosity of 306i) mPas. {@6329} nre: 1, Part A: Disperse Besifloxaein in glycerin and deionized water in the main vessei and heat to 700C. b.) . Part B: Heat in eetyl Alcohol, light liquid paraffin, cyclopentasiloxane, steareth 2, and steareth 21 and in a separate vessel to 790C. 3, Add PART B into PART A with continuous mixing at 760C and allow to mix for l5 min. Cool the batch with. mixing to 45°C.

U1 4. PART C: Swell the earbopol separately in water for 2h . Add PART C into PART A/B and mix well for El) min.

Example 2i: Minimum inhibitory Concentration of ln-ilouse Besiiloxaein Gels (1%) and its Combination with Adapaiene (9.1%) {$933M Method: Minimum inhibitory tration. of the test gels (’i‘able is, ll} GL1 and GL2) are determined by micro broth dilution method against P. genes MTCC 195] (strain susceptible to Clindamycin). Bill broth and Bill agar media were prepared as per the manufacturer’s instruction and autoclaved at l2l.OC for l5 s. P. aortas culture is grown in Brain Heart infusion agar (BillA) at 37°C for 48h under anaerobic condition. For MlC determination testing, gels are dissolved in the t and r diluted with Bill broth. Then, 96 wells plate are filled with lOtlul of Bill broth containing drug with different concentration to get the final concentrations of {106, (3&3, 0,255 9.55 l and Zug/ml in ent lanes (lane l to lane 6). Remaining lanes of the 96 well plate are used as growth control and ity control. Finally, P. genes culture suspension (approx l.5xll)6) is added in all the wells except sterility control wells and plate-is incubated at 370C for 48—72h under anaerobic condition. At the end of 72h, Alamar blue solution (Emil) is added into the wells and incubated at 37°C for 2h. Plate is visualized for ial inhibitions and MIC values of the tested samples are determined. Gel formulation containing Besifloxacin alone? Besiiloxacin combination with adapalene and their placebo are analyzed for lVliC determinations and results are shown in Figure 6.

Eliti33i} Results: Minimum inhibitory concentration (MlC) assay showed that MIC values of the besiiloxacin in both the ations (GLl and GL2) were found to be similar in the range of ii. Bug/ml to b.25ug/rnl {Figure 6). 3t) Example 22: lines Response Curve (using Zones of inhibition) of Gel containing liesil'loxaein alone, its ation containing adapalene against P. cones {@332} Agar well—diffusion method is employed to run Zone of inhibition (ZQl) assays. ZOl is employed to assess the potency of formulations (Tablet, GLl and G12) consist of Besifloxacin alone and its combination with adapalene to inhibit the growth of microorganisms under study. ZQl , determined at different APl concentrations, can be used to derive dosenresponsecurves (DRCs) for efficacy ison of different formulations. {@9333} Method: P. genes cultured in Brain l-leart infusion (Bill) Broth (37°C, 48h) under appropriate ion to get the desired CFU count, to be used to l0 inoculate the plates. TSA plates are spreaded with lililul of 0.5 McFarland equal bacterial suspension. Sterile disc {6mm} are loaded with various concentration of gel formulations (equivalent to different Besii‘loxacin concentration of 0&2, 0.25, 0.5 and lug/ml) and/or controls ( ltBOnl each) and then disc has been placed above the spreaded , Thereafter, the treated plates are incubated at 379C for 24h, l5 Readouts are taken after 24h and effect of combination of two different Af’ls on cne activity is measured using Zone of inhibition studies. {($334} Result: ZQl assay results showed that both the ations have similar anti~acne activity as evident from their zone of inhibitions. Adapalene presence in the gel formulation is not affecting the anti—acne activity of the Besifloxacin present in the formulationstFigure 7).

Example 23: ’l‘ime Kill Kinetics Evaluation of Gel Containing Besilloxacin alone, its Combination ning ene against P; cellar Etill335l An activity comparison, by in vitro tinte~hill kinetics of anti~acne agents using besifloxacin gel (Table 18, GL2) and versus its combination with adapalene gel {Table 18, GLl) has been demonstrated. The time—kill assays are used to te efficacy of antimicrobial agents, either single or in combination. 3t'sl Method: P. acnes are suspended in brain heart infusion broth (Bill broth) at inoculurn concentration of l.3>c:ltl8 cells/ml. Cells were taken from a freshly growing (3—7 days old) plate and cell suspension is vortexed to remove the cell clumps as much as possible. The media is then supplemented with appropriate trations of gel formulation {equivalent to l gag/ml and lOug/rnl of Al’l) in the reaction mixture. The cultures are incubated on a tube rotator at 37°C in anaerobic condition for 2h, Sit and 24h. At the end of each time points, aliquots (Sfiul) of P. cones cultures are ly diluted with medium and plated on brain heart infusion agar plate. The plates are incubated at 37°C in C02 incubator for 3 Uh days. The viable colonies are counted and converted to CPU/ml. The s of time kill study using oxacin gel. stand alone and its combination with adapalene concentration are plotted in Figure 8. {$9337} Resultszl‘ime hill, assay results showed that both the formulations have similar kill kinetics against P. genes. Presence of adapalene in the gel formulation l6 did not appear to affect the bactericidal activity of the Besifloxacin present in the formulations. Both the formulations also have concentration dependent hill kinetics at two different concentrations of lug/ml and ltlug/ml e 8). P. acnes’s lrill, has not been observed with the placebo gel? which indicates that o gel is not imparting any anti-bacterial activity. l5 Example 24: in vivo Time Kill Kinetics of Gel €ontaining Besitloxaein against elindamycin resistant R genes. {93338} An activity comparison in in viva time~l<ill assay of anti—acne agents using besifloxacin gel and versus placebo gel has been performed in mouse model. This assay is used to determine efficacy of formulation containing 2% antimicrobial agents to kill the pathogen infecting a live animal: 39l Method: Clindamyein resistant P. cones cells were grown in brain heart infusion broth (Bill broth) till the cells reached late log phase of growth.

Cells were washed twice and resuspended in BHl broth with final inoculum tration of 2xl07 cells/ml. Cell suspension was vortcxed and passed through 3th.} s to remove the cell clumps as much as possible. l0 ul of P. cones culture were injected into right ear (dorsal surface) of anesthetized gull) weeks old mice by the help of Hamilton syringe (intra dermal ion). After 3t) minutes approximately l5 mg of l% Besifloxacin or Placebo gel formulation was d on mice right ear (dorsal surface) & properly ed with the help of spatula, After 24 hrs. Mice were sacrificed and ears were ted (0 hrs. Control mice car had been taken on the same day when P. cones were injected) and placed in microeentrifuge tube 1 ml of Bill broth added in each tube and then ears were nized by mechanical nizer. 50 iii of ear homogenates from each tube were elated on BHA plate (containing 0.5 ng/ml Anthotericin—B) after serial dilution. Aliquots (SOui) of P. acnes cultures were serially diluted with medium and plated on brain heart infusion agar plate. The plates were incubated at 37°C in C02 incubator for 3 days. The viable colonies were counted and converted to CFU/ml. The results of in vivo time kill study using Besifloxacin gel vs placebo gel were plotted in Figure t}. ltltlfidtil Results: in vivo 'i‘ime kill assay results showed that Besifloxacin gel it} ation have the ability to clear almost L5 log CFU (~95%) of inoculum of ciindamycin resistant P. genes within first 24 hours (Figure 9). There were some nominal killing being observed even. with the placeho treatment it could largely be attributed to the immune competency of the host it fiirther vindicated our claim that not only in in vitro but also in an animal infection model our topical ations were very effective and could penetrate at the site of infection in sufficient quantity and clear clindamycin resistant ion.

Example 25: Freparation of Spray Formulations Loaded with Besifioxaein iriytirimhiorides tilinafloxaein or Sitai‘ioxaein Alone and the Combinations with Adanalene lflfi341§ Soray formulations containing hesifloxacin Hydrochloride, eiinafioxaein, sitafloxaein and combination with adapaiene and salicylic acid are formulated as per the itions shown in Tahle 1?. These formulations have the pH of The formulations consist of actives (hesifloxacin hydrochloride, elinafloxacin and sitafloxacin) equivalent to i%w/w in different formulations (Table lit, S5, 33 and SE). in addition to stand alone formulation, anti-microbial agents are combined with keratolytie agent such as adapalene (tilt/o) to provide both the anti-acne and keratolytic activity in ts ing from acne (Tahie 1?, Sit S4 and so). $39342} Method of preparation: (1) Water is added to a main mixing vessel followed by the additien 0f sodium hydrexide on, PEG MSG, methyl. h—fZO and glycerin in order with mixing. (2) in a separate vessel, hesiiloxacin hydreehleride, eiinafiexaein er sitefiexaein is added in isepropyl aleehel, ene glycol, diethylene glycol monethyl ether and ethanol mixture and mixed. Ali the content of this vessel is added to the main vessel and mixed. palene is disselved in N-methyl 2~pyrrelidene and added to main mixing vessel, and mixed. ii) (4) Salieyelie aeid, phenexyethanel, and sodium hydroxide is added te adjust the pH te 5.0. (5) Fragrance is added to the stirring mixture and stirred eentinueusly tili uniform mixing. -:_— 225 -25 VVVVV -—-—-1.5 diethylene glycol monethyi ether Example 2d: Preparation ei‘ Face Wash Fermulations Leaded with Besii‘ioxacin hydrochleride, Clinai'inxacin er Sitai‘lexacin Alone and the ations with Adanalene 396343} Faeewash formulations containing oxacin, clinafloxacin, or sitailoxacin and the combination with ene or salicylic acid are formulated as per the compositions shown in Table Ztl. These facewash formulations have pH of 4.7—6 and viscosity of around 1500-59th m?a.s. The formulations consist of. s (hesitloxacin hydrochloride, clinaflcxacin or sitai‘lcxacin) equivalent to l%w/w in different formulations (Table 2t}, FWS, Fifi/’3 and FWZ). in additicn to stand alone formulation, antiumicrchial agents are combined with olytic agent such as adapalene ((1.1%) to provide both the anti—acne and keratolytie activity in patients suffering from acne (Tahle 2t}, FWl, 13in and FWd). {@3344} Methcd ei‘ prenaratian: (l) in a main mixing vessel, water is added. Then carhopol aqua SF-vi is added slowly at a low speed (70-89mm) of mixing. (2) in the same , sodium C1446 olefin sultbnate (40%) and scdiuin lauryl ether sulphate (28.6%) are added while stirring. (3) The mixture is neutralized with sodium hydroxide, adjusting the pH to 6.5 to 77.6. The mixing speed is slightly increased to ensure m mixing. 2t) (5) Then cccamidopropylhetaine is added to shove rnixnire with continueus stirring followed by slow additinns of disodium EDTA and glycerin. (6) exacin hydrochloride, clinalloxaein, or sitai‘loxacin and adapalene are added to the above stirring main vessel. (7)Adapalene is dissolved in N~methyl Z-pyrrolidcne and added to main mixing vessel, and mixed. (8) Then, salicylic acid, propylene glycol and PEG—7 glycerylcceoateare added to continuous stirring main . {8) Finally, pill is adjusted to 5.5 by the addition of citric acid, Viable 2t}: Fnee wash for compositions FWL FWZ, FW3, FWé‘t, FWS anti FWé (Joniositlon ~ % 8. Na. ingredient ; l Winn 31.3. 2 Carbo 01an SF-l 6 Sodium (314le Olefin 3 t g 35 Sultonate g 4 Sodium lauryl ether suighate ““2; a 5 : Sodium ide(18% a.) qs, .. .. .. .. : g 6 eccenttdegrogyibetaingflint) 10 _- in l0 5 m; , _____Zml 7 DisodinmEDTA 0.1 0.1 0.1 0,} n1 cs: 8 Glycerin l 9 Besifloxacin Clinafioxacin i l. l Sitafloxacin , Adapalene 16 ; V Citric Acid(50%) Example 27: Preparatinn nt‘ Soap Bars Limited with Besit‘ioxnein liytirociiloritte, Clinafioxeein or Sitnfioxaein Aione and tire Qombinatione with Adepaiene 5} Method of preparetinn: Soap bars containing besifloxacin, clinafioxacin or sitafloxacin are formulated as per the compositions shown in Table 21. The soap bars t of oxacin hydrochloride lent to 1%w/w (Pattie 21, S85), ciinafioxacin equivaient to 1%w/w (Tabie 21, $83), sitatloxacin equivalent to 1%w/w (Table 21, S82). in addition to stand alone it) besitloxacin formulation, besitloxacin is combined with atiapalene (0.3%) (Table 21;, Sal) and salicylic acid (Table 21, $86) to provide both icrobial and keratolytie activity to patients suffering from acne. Similarly, another formulation contains clinafioxacin in combination with adapalene (Tania: 21, 384). {$3346} Method of prenarntion: (l) Sodium palrnitate is blended with ing ingredients in the mixer. (2} The mass is passed through the roll mill and plodder followed by billeting and stamping at a temperature between 35°C—40°C. iTahle 21: Sean bars fer cemnesitions Slit, $32, $33, $34., and 33% Comositinn 2%: Fibrin ingredient lanryl sulohate _______2“mlmfgodinm L3 Polyqna'temium-39 ___ 4 g Methyl GlucethQG Titanium dioxide 0.5 05 a BHTiBml/lmd act act ant 0.01 : Hydroxylolnene) Example 28: a’tion of Body Wash Centaining Besifloxacin Hydrochlnride, {Illnaflexacin or Sitafloxaein Alone and the Cnmbinatiens with Adapalene {@3347} Method of atien: Body wash formulations containing it} besifioxacin, clinalloxacin or sitafloxacin and adapalene or salicylic acid are ated as per the compositions shown in Table 22. The body wash formulations consist of besitloxacin equivalent to 1%w/w (Table 22, BWS), sitalloxacin equivalent to l%w/w (Table 223 BWZ) and clinalloxacin equivalent to l%w/w (Table 22, 3W3), in addition to stand alone besifloxacin fortnnlatien, l5 besifloxacin is combined with the adapalene (G.l%) and salicylic acid (2%) (Table 22, EW’ l in acne , BWé}, to provide both acne and keratelytic ty patients, Similarly; another fonnulation contains clinafloxacin in combination with adapalene (Table 223 8W4).

Tahie 22: Eddy Wash fer Cnmpnsiiiens 3W1. 3W2, 3W3, 3W4, 8W5 and iiinnW%} Ammonium ianryi sul hate (30%) Prepyi harahen Methyl iueeth-EG methmimmg q E63348} Methed nf prenaratinn: U1 ( i) in main mixing vessel, nni EDTA is disseived in water. (2) Carbpe]. aqua SR1 is added to the main vesgei. (3) Then it is stirred for 10 min feiiowed by the addition of ammonium iauryi sniphate. (4) Then propylene glycei, hesifidxaeiii hydreehldride, eiinaflnxaein, or siiafloxacin are added to above mixture with continudns stirring. (5) Adapaiene is disseived in N—methyl 2~pyrmiid0ne and added to main mixing vessel, ed by its mixing. (6) Whiie stirring remaining ingredients are added to the above mixture. (7) Finaiiy, neutraiization is done with iriethanoiamine and mi is ed to ES 5.5-6.0.

Example 2‘}: Preparation of Lotion ations Loaded with Bosifioxocin Hydrochiorido, Ciinafioxacin or Sitai‘ioxacin Alone and the Combinations with Adanaiono 9} Lotion t‘onnolations containing besitloxacin are formulated as per tho ‘Ji compositions shown in Table 23. These s have pH of 4.7—5.5 and viscosity of around 251111430130 mPa.s. The lotions consist of bcsifloxaoin oquivalent to 1%w/w (Tania 23, L5), sitafloxacin couivalcnt to 1% w/w 'i‘abio 3, 142) and ciinafloxaoin oooivaiont to 1%w/w (Tobie 23, L3), in addition to stand alone formuiation, bcsifloxacin is combined with tho adapalonc (0.1%) (Tobie 23, Li) it) or salicyiic acid (Tania 23, L6) to provido both thc anti—acne and kcratoiytic activity in pationts Suffering from acnc, Similarly, another formulation ns clinafloxacin in combination with adapalonc (Tobie 23, 144).

{Table 23: Lotion Formniations for Compositions L1, L2, L3, L4 and L5 11 , , ion.

Comosition (0/0 L3 L4 L5 to _ VVVVVV L 1 Rigor tits ggw l 2 Disodium EBTA o5 aos 3 1 1 ______ Qatbopol aqua SF-t 1. 1 ________g atum _ C'clomettiioonc 0.5 {LEW a 6 Sofbitan stoatato - . . 1.4 7 60 . , 0.6 (1.6 . __Polysorbatc m V a Methyl giucah—zo 1. l 1 1 1 9 Cetyi alcohoi 1.6 1.6 1.6 1.6 1t} Tocohcryl acctatc . , V0.25 0.25 11 1 1 _____ aesinoxggin __l_ ___g ___Q C1i_n_atloxacin__ o o o l 13 Sitatloxaoin t) 17 Pro ylono glycoi 2 VVVVVVV 18 __olvcean 8 L 19 Ethanol 2 2G Phonoxyothanoi a . . g 1 '21 Sodium hydroxide q.s q.s as as as .j {@9356} Methud uf nrepara‘tien: (1) in main mixing vessel, disedium EDTA is disseived in water. When it is fuiiy disseived, mixing speed is set at 160 rpm. (2) Then, earbopei aqua SF-i is siowiy dispersed irate water and stirring is continued until cempiete mixing. (3) Heat the mixture to 70°C. (4)Meit petrelatumg eyeiomethicene, peiyserbate 60., serhitan strearate, eetyi aicehei in a separate beaker and add it to abeve mixture. Keep the stirring speed at 290 rpm. i0 (5) Aiiow the mixture to cool at 35-40% with nt stirring at 2% rpm. (6) Add tocopheryi acetate tn the above mixture after it reaches at C. (7)8esifiexacin hydrochioride, eiinaflexacin or sitai‘ioxaein is added to the abeve mixture by sing in water. (8) Adapaiene is dissoived in N~methyi Z-pyrreiidene and added to main mixing vesseig t‘eiiewed by its mixing at 250 rpm. (9) Eisseive saiieyiic acid in ethanei, propylene giyeei and giyeerei and add it tn main mixing vessel. (10) Add phenexyethanei t0 the above mixture with nt stirring at 250 (i 1) lize the wheie mixture with sodium hydrexide and mix for 30 min.

{MESS}; Exampie 28 and Tahie 24 be some exemplary femuiatien comprising seuhiizied API, (eg, besifiexacin hydroehieride).

Examnie 3t}: Appreaehes used fer the seiuhiizatien {if exaein Hydruchieride {33352} Surfactants are known to seiuhiiize the hydrephohic nces by reducing the aeiai tensien. fin—addition to surfactants. ee—soivents er ee— tants aise helps in seiubiiizatien of the peerly water-seiuhie cempeunds by increasing the wetting preperty er reducing the interfaciai tensien of the hydrophobic meiecuie. in this patent, hesifioxacin have been soiuhiiized using at) surfactants such as sodium iauryl suifate, tween 80, tween 29 and span 8t), and coseivents/eesurfactants in the deiivery vehicies. The presence of the sodium lauryl sulfate greatly enhanced the aqueous solubility of the liesilloxacin hydrochloride and used for preparation of topical formulation (Table 24, SClg 8C2 and SCSLTl‘re CG-SOlVinS such as propylene glycol monoeaprylale and dielhylene glycol rnonoelhyl ether has been used for the preparation of cream formulations (Table 24, 3C3 and 8C4).

Table 24: Fnlly Solnlilllzerl beell’loxeeln ereern formularlnns S. No. Composition (1% wlw) ingredients mmmme...” . ser__j_ Sez “em _SC4 see l Besilloxaein hydrochloride 1.99 1.09 1.99 1909 E 1.99 %___________J.....www.mummww _______ : 2 Sodium Lauryl sulfate i..._...__.......‘._.1.........

Cetyl alcohol Propylene Glycol Dierliylene glycol mono~etliyl ether n Stearetlr-Zl :2 12 Cyclopenraslloxane 4 G 4 3 8 13 y ethanol 0.5 (2.5 0.5 0.5 9.5 14 Sodium relycimxide {10% in water) l r l r 2 l5 lfijarbomer homopolymer type C 0.29 0.80 (HS 0.8 G ; 16 ol Ultrez l0 l) l) 0 0 63.3 17 Water :1, so q. s. q. s. q, s. q. s. i {$6353} es 3i and 32, and Tables 25 and 26 describe some exemplary formulations comprising suspended Al’l.

Example 31: Preparation of Suspended Brag Loaded Gel Formulations with Minimal Soluhilization-repreeipitation of the firing {99354} Preparation of oaded {suspended form) gel via a conventional method usually leads to exposure of the drug to a wide range of pH conditions, which may lead to, in some instances, solubilization of the drug, and then reprecipitation. This soluhilizationwreprecipitation phenomenon in most cases leads to change in original particle size, ty profile or crystal pattern, or others. As an example, hesiiloxacinllCl (which shows pll dependent solubility) displays this solubilizationwreprecipitation phenomenon, where pit of about 4.5 or it) below solubilizes the drug to a significant and variable extent. The solubilized besifloxaein then? upon se in pH (during formulation preparation), reprecipitates, which may result in one or more unwanted changes. {@355} in order to circumvent this issue? a modified approach has been employed to prepare different suspended drugmloaded formulations. Table 25 and the Method of Preparation below details the gel composition and preparation with negligible or minimal drug solubilization—reprecipitation. These gel fonnulations have off—white ance with pH of 5.0—6.0 and approx. viscosity of about 3900 to about 5500 mPais measured by eter (RheolabQC, C—L’l‘l) Sill/QC, Anton Paar} The formulations are used against tible and resistant acne conditions.

Tahle 25: oxaeinHCi Suspended Gel Formulations for €ornpositions GL5, one, GL7 Chemical Name Composition {%w/w) l--§esimmfld;<acin.lit3l Equivalent to l l5 2 Besifloxaein Allantoin 0.2 0.2 0.2 l-Carhomer homopolymer type C“ 0,85 0,85 ether ' 5 o V VVVVVV Edetate disodium dehydrate (ll. 6.1 {3.1 Phenoxyethanol (3.? (if? (3.? Polyethylene glycol 490 ‘_ 5 5 Sodium hyaluronate -0.4 (2,4 9,4 Sodium hydroxide on* qs. to adjust q.s. to qs, to adjust pH adjust pH oil Purified water qs. q.s. qs.

Phase A: Purified water, Edetate disodium dehydrate, Allantoin, Carbomer homooolymer type C Phase B: Purified water, Sodium hyaluronate Phase C: Phenoxyethanol, Sodium hydroxide solution Phase D: Glycerin, BesifloxacinHCl (Equivalent to Besifloxacin), Purified water, Sodium hydroxide solution Phase E: Polyethylene glycol 4th), Diethylene glycol monoethyl ether, Phase F: Sodium ide solution ll} {$51356} Method of l’repara‘tion: l) in a main mixing , edetate disodium and oin were dissolved in water. Then oarhomer homopolymer type C and hyaluronate sodium were added and allowed to swell at 200 rpm for 60 min. Then phenoxyethanol was added to the carbomer mixture. Then, pH of the mixture was raised to l5 6.9 with sodium hydroxide solution. 2) in a separate vessel, glycerin and hesifloxaein.liCl were dispersed with continuous mixing at 390 rpm for it) min. 3) Dilute solution of sodium hydroxide was added drop~wise to the separate vessel to adjust oil to 5.5. 4) The contents of the above e were added to the main mixing vessel with stirring at 20% rpm for 2 hours.

) Finally, hylene glycol and diethylene glycol monoethyl ether were added to the main mixing vessel and mixed for further 20 min. 6) White~to~pale yellow gel was obtained.

Exampie 32: Preparation: nf Suspendeti Brag Leaded Cream Farinuiatiun by Avniding Re-preeipitatien {if Brug §9G3S7§ Similarly, cream fonnuiations containing suspended besit‘iexaein were ed, with minimai soiubiiization—repreeipitatien, as per the itiens shown in Tabie 25. These cream atiens have offuwnite appearance with pH of 5.0u6.(3 and . viscosity of about 3009 to about 4000 rnPas (measured by Viseometer (RheelabQC, C—LTD gO/QC). Anton Far). The fonnuiations were then tested against susceptible and resistant strains of acne. "faith: 26: Besifinxaeinfifii Suspentieti Cream Fermuiatiens fer {Zampnsitiens {SR/i2, CM3 anti Cit/i4 ingretfiienta €empesitien (”/8 wlw) “mm ‘ ext/i“? “m“mi Besifiexaeinflfl equivaient t0 1.5 i 2 besifluxaein 0.1 {IL} 0.1 a Butyiated Hydrexytoiuene Carbopel nan (2%) 3n Cet‘yi aleehei 1 i i W5 5 5 5 Light iiquid paraffin 3 3 3 Pitenexyethanoi 0.5 “m5?” WWW Sodium nydmxide seiution 2 2 2 i 1 nmmwmm i 0 t Steareth 2 i Steareth 21 2 i i Steai'yi alcohoi ‘ Purified water (5.5. t0 10G q.s. tn 100 qa. t0 109 Phase A: Cyclomethieone, Span 8t), Cet‘yi alcohoi, l alcohol, Light liquid paraffin, Steareth 2, th 2i Phase B: Glycerin, Besifloxacin.HCi equivalent to besifloxaein, d water, Sodium hydroxide solution Phase C: Butyiated i-iydroxytolnene, Phenoxyethanoi, Carbopol 98d (2%) ETTSTSSSE Method of T’reparation: l. in main mixing vessel: hesiiloxacin was dispersed in glycerol and water with uous mixing at 3th rpm for 10 minutes 2. Dilute on of sodium hydroxide was added dropowise to in l0 mixing vessei to adjust pit to about 5.5 and heated to 70°C 3. in a separate vessei, cyciomethieone, span 80, cetyl alcohol, stearyl alcohol, light liquid paraffin, steareth 2 and steareth 2i were heated together to 70°C. 4. Heated mixture of oil phase was added with. continuous mixing to the i5 main mixing vessel at 70%; 2G0 rpm and allowed to mix for 15 min.

. The content of the main mixing vessei was allowed to air—cool with mixing to 45°C. 6. Carhopol was allowed to swell in water for 2 h and its pH was adjusted to about 5.5 to 6 with sodium hydroxide solution, which was then added to the main mixing vessel and mixed, 7. Remaining components butylated hydroxytolnene and phenoxyethanol were added to the main mixing vessei and mixed for 29 min. 8. The cream’s pH was adjusted to about 5.5 to 6.9 with sodium hydroxide, if required: Exampie 33: Preparation of Formulations Containing Combination of Actives (soinhie antinmierohial and suspended keratolytie agent) {@359} Gei formulations containing a combination of solnbie hesifioxacinHCl and suspended adapalene were prepared as per the itions shown in Tahie 27‘ These gel ations have pale yellow ance with pH of around 4.5 and viscosity in the range of 3000 to 5600 mPas (measured by Viscometer {RheolahQCg C-LTD Sill/QC), Anton Paar). The formulations containing soluble besifioxacin equivaient to 1—2% w/w and iy 0r fuiiy suspended adapalene equivafient {Q (11% w/w have bgen prcpared t0 e anti-acne, keramiytic and anti—inflammatery effscts in patients afiected fmm susceptibie and ant acne.

(J: Tabie 27: Besifiaacimflfl (gamble) and Adagiaiene (susgended) Centafining Gei Farmuiatéflns {9r Campasétiflns SL1, SL2 amfi SL3 [ Engreéientg Camgwsiéien (% w/w) SL2 g SL3 BesifloxacinHCI iem to Besifloxacin Adapalene Alianmin Diéthylené glycol monoethy}. I E ether Edstafic disodium dihydrate H4. G. 1 gig/“Ta:“““““““““ 5 Hyaiumnate Sodium 6.3 Q 9.3 E-{ydmxy cthylceiluiosg (l9 1‘2 0‘9 iPoloxamsr 0.2, 6.2 0.2 MHWW‘ 7 ;Purified water (5.5.30 (1,3. to q.s.t0 E00 E 109 £90 Phase A: Purifiedwater, Aiiantoin, Edema: disodium dihydrate, Hydrexy ethylceilulose Phase B: Purified water, Hyaiuronata Sadium Phase C: Purified water, Glycsrin, BesifloxacinHCl Equivaient to xacin, Adapaiane, Poioxamsr Pelyethyiene glycai 400, Diethyiene giycoi mamathyi cther, Phsmxyethanoi. ltlllitdlll Method of lireparation: l. in a main mixing vessel, edetate disodium dihydrate and allantoin were ved in water at 400 rpm for about it) minutes. 2. llydroxyethyl cellulose (HEC) was added in portions to the main U} mixing vessel at very high speed (about 406—5th rpm). 3. EEG was allowed to swell for l h with stirring at about 100450 rpm. 4, in a separate vessel, hyaluronate sodium was added and d to swell for 15-30 minutes, followed by addition to the main mixing vessel. it.) Us in a separate , glycerin and besifloxacinHCl were dispersed in water and mixed with glass rod. Then, diet’hylene glycol monoethyl ether and polyethylene glycol dill) were added to oxacin dispersion and mixed. This dispersion was added to main mixing vessel, and allowed to stir at about 0 rpm for about 5 minutes. l5 6. Phenoxyethanol was then added to the main mixing vessel, and mixed at l00~l50 rpm until any polymer lumps disappeared completely and clear gel was ed. 7. Adapalene was dispersed in aqueous solution of poloxamer and added to the main mixing vessel, resulting in white~to—pale yellow opaque gel 2% containing soluble besitloxacin and suspended adapalene. liltifédll Example 34 and Table 28 describe some exemplary formulations which are essentially free of a ning polymer.

Example 34: Preparation of Suspended Brug Loaded Cream ation without use of polymer as a viscosity modifier {9&3le According to published literature there may be some kind of physical and / or chemical interaction between Carbomer and iluoroquinolones. For which, there may be a need to prepare formulations without Carbomer or er - olymers to avoid any incompatibility issues during the product shelf life.

Towards this, an alternative cream formulation, without use of the Carbomer or any other polymer, has been prepared. Cream formulation compositions and procedure is given in Table 28.

Table 28., BeslfloxaeioHCl Sesgrerirtetl Cream Formulations for Compositions (Ill/ES lulngredieets Composition ”(7% 1 Besifioxaein Butylatetihydroxy toluene M—flmmwi Cetyl alcohol Cyclopentasrloxane ' ‘ Glycerin Mineral on w Phenoxyetharrol 0.7 2 Stearyi l Polyoxyl Ether yl 2l Stearyl 2 Ether l alcohol 2 Sodium hydroxide 01.3. to pH > 5.5 red Water_______._____._._.___._____._.._.________,.+___.Mars. to 1000 ”mi (1} Phase A: Cetyl alcohol, Stearyl alcohol, Mineral Oil, Cyclopehtasiloxahe, yl 2 Stearyl Ether, Polyoxyl 21 Stearyl Ether Phase B: Glycerin, Besil‘loxaeio, Phase C: Sodium roxiriea Purified Water Phase D: Butylatedhydroxy toluene, Phenoxyethanol l0 Phase E: Sodium hydroxide {00363} Methotl of Preparation 1) Phase B: Glycerol and API were mixed together in main mixing vessel? 2) Phase C: Sodium Hydroxide and water were mixed together and added slowly into phaee B in the main mixing vessel and the contents heated to oil—65°C, 3) Phase A excipients were mixed together and heated at 60~65°C, followed by adding to the main mixing vessel with continuous overhead stirring at about 600 4) The contents of the main mixing vessel are then allowed to cool to about 400C, ) To this mixture contents of Phase l) were added and mixed for l5 min, 6) Finally pH of the formulation was ed to 5.5 - 6.0 using phase B {@364} Exampe 35 and Tables 29—32 describe some exemplary formulations sing different polymeric or non—polymeric viscosity modifiers or gelling l0 agents Example 35: Use of Different rs for Viscosity Regulation of Formulations Containing Solubilized Besit’loxacindfiCtl‘lZ} } The purpose of gel formulations to be prepared was to have acceptable viscosities, while the active drug remains in soluble form. This becomes l5 challenging, when the pH required for drug solubilizing or stabilizing a solubilized drug is outside the normal range of about 5.0 to about 7.0 for topical products. For example, besilloxacinHCl goes in solution, of course with proper choice of excipients, when the pH is adjusted to below 5.0, for example in the range 5. Not many rs (without affecting gel sensorial parameters) 2% were found to be able to furnish acceptable viscosities to the gel ations, in this pl-l range. {@9366} Eifferent polymers (and their different grades) like carbomer, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, sodium hyaluronater and other polymers were used to prepare gels with acceptable viscosities, wherein the active drug was desired to be in a solubilized state. liltiiiti’l'l Gel formulation ning soluble besifloxacinHCl was attempted to prepare using Carhomer as per the composition and ure given in Table 25.

Although the drug could be solubilized at pH 4.5, the Carbomer used in required amount was not able to impart acceptable viscosity to the formulation. The resulting formulation had viscosity about 1800 ml’as (measured by Viseometer (RheolabQC, Gill) SG/QC). Anton Paar).

Table 29. exueindfitfil Seluhilized Gel atlen Prepared Using Carhemer fer Cempesitlens SL4 ingredients Campeeltlen (0/?) wfw} Glycerin BesifluxaeinHCl equivalent to l Beslfloxaein B Purified Water cyst t0 liltlll Carbomer Homepelymer Type C Allantein Edetate Disodium Dihydrete ill (EDTA) timed“?““““““WWM Sodium l-lyaluronate C Polyethylene glycol 400 6 Dieth‘ylene glyeel rnenoethyl ether ll ltlll368} Methed et’ preparatien: l) Phase A: Glycerol and besifloxaein were mixed together in a separate vessel, 2) Phase B: in a main mixing vessel, allantein and EDTA were seluhilized in water with stirring at Zilll rpm, then earbomer was sprinkled over it slewly and allowed tn swell for 45 min, ll) 3) Sodium hyaluronate was sprinkled into the above e and allowed to swell for 15 mini followed by addition of Phenoxyethanol and mixing, ) Phase A was transferred into the main mixing vessel with continuous ng at 209 rpm, and mixed for 30 min, 6) Polyethylene glycol 409 and diethylene glycol meneethyl ether were added t0 p...“ “J: the main mixing vessel and mixed for l5 min at 156 rpm {@369} llydroxypropyl Cellulese and yprepyl Methyl ose are widely used in oral and tepieal pharmaceutical formulations and available in number of different grades that can furnish wise viscosity ranges. Formulations ning soiubiiized besifiexaein were prepared using hydrexypropyl eeliulese er hydrexypmpyi methyl celfiuiose as viscosity modifiers (Tame 3%). Aitheugh aceeptabie viscosities of about 300% mPa.s {measured by Viscometer (RheoiabQC, Cdfiffi) SEE/QC), Anten Paar) were observed using both the LA poiymers; the senseriais were not acceptable.

Tabie 3i}. Besifiexaeinfifl Seiuhiiized GeE Fermuiafiefls Pregared Using Hydrexypregy! {ieiiuiese and. Hydrexypmpyi Methyl Ceiiufiese fer (Iempesitiens SL5 and SL6 ; Phas ingredients Cempesifiens e (% wlw) SL5 SL6 A ein xaeinfiili U39 1.09 B Purified Water q.s. to EGG!) q.s. t0 IGGQ Eiydreg§§¥5§§i€i§fifi§éémmmmmm ' g 1.5 e ein {)2 0.2 Edetate Disodium Dihydraie {11 6.1 (EDTA) 1 1 LEWEfiEF"‘‘‘‘‘WT“ 9.7 “T“ Sodium Hyaiumnate 0.4 (3.4 C Peiyethyiene (Eyes! 4% 6 6 "egegegzgeéaieeeafiggee?7“““ """" 11 11 {$93M} Methed ef E’regaraiienz 1) Phase A: Glycerin and besifloxacin hydreehleride were mixed tegether in 3.

Separate vessei, 2) Phase B: in a main mixing vessel, allantoin and Edetate Disodium Dihydrate were solubilized in water with stirring at 260 rpm, then hydroxypropyl cellulose or hydroxypropyl methyl cellulose was sorinltled over it slowly and allowed to swell for 45 minutes, “J1 3) Sodium hyaluronate was sprinkled into the above mixture and allowed to swell for l5 minutes, followed by addition of phenoxyethanol , ) Phase A was slowly added into the main mixing vessel with uous stirring at 2009 rpm, and mixed for 30 min, 6) Polyethylene Glycol 400 and Diethylene glycol monoethyl ether were added l0 to the main mixing vessel and mixed for l5 min at lSG rpm {993%} Hydroxyethyl cellulose is another widely used excipients in oral and topical pharmaceutical formulations and ble in number of different viscosity grades. Formulation containing solubilized hesifloxacin.llCl was prepared using hydroxyethyl cellulose as viscosity modifiers (Table 3E). Using this r, the l5 prepared gel could display good sensorials, at pH of about 45, along with other parameters like acceptable viscosity and soluble drug.

Table 3]. Besil‘loxacin.ll€l Solohilized Gel ations Prepared Using WWW... ients Composition (% wlw) BesifioxacinllCl Allantoin Diethylene glycol monoethyl ether Glycerin 5 5 5 oxy ethyl cellulose Phenoxyethanol Polyethylene Glycoléitltl ‘ Sodium hyaluronate Purified Water ‘ a. s, to ltltl q. s. to ltltl or s, to lllt) Phase A: Purified water, Allantoin, Edetate um Dihydrate {EDTA}, l-lydroxy ethyl ose Phase B: Purified water, Sodium hyaluronate liliase C: Purified Water, Glycerin, BesifloxaeinllCl, hylene Glycol 400, Diethylene glycol monoethyl ether, Phenoxyethanol Method at” lhreparatinn 1) In a main. mixing vessel, EDTA and allantoin were ved in. water at 400 rpm for ll) min, 2) Then, hydroxy ethyl cellulose was sprinkled to the main mixing vessel at very it) high speed (around 409-500 rpm) and allewed to swell for 1 h at 109450 rpm, 3) ln a te vessel, sodium hyalumnate was taken and allowed to swell with water fer 15- 30 minutes. Then, added to the main mixing vessel, 4) in another vessel, glycerin and besifloxaein HCl were dispersed in water and mixed with glass rod, To this dispersinn, diethylene glycol monoethyl ether and polyethylene glycol 4% were added and mixed, } Above dispersion was added to the main mixing vessel, and allowed to stir at lthl- lSt) rpm for 5 min, 6) Finally, phenoxyethanol was added to the main mixing vessel, and mixed at lGG—lSG rpm until polymer lumps disappears, if any and clear gel is obtained. 26 Examnle 36: l’reparatien at" (Eel. Fermnlatlnns Leaded with Bifferent ntratiens {if Suspended Besifiexaeinfifil tn Ghserve the Effeet on Visedsities at Fermalatinns Cnntaining Hydrnxyethyl oseflil] lllll372l Gel fermulations containing different concentrations of hesifloxaein hydrochloride were formulated using hydroxyethyl cellulose as thiekening agent as per the itions shown in Tattle 32.

Table 32: Gel Formalstiens with fiifferent Ceneentratinns at“ Suspended Besillnxaein.ll€l Using Hydrexyethyl Cellnlese fer Compnsitiens GLilg, GLlll, GLll, GLEZ and GLB Chemieai Name Cempositien 51% wiw) {31.41%} GL1} GLEZ GLES Besifiexaein.H(31 (Equivaient t0 0 4‘ 8 Besifiexaein) E Ei-Aiiantoin 0.2 Diethyiene glyeoi menoethyi ether Edetate disediurn dehydrate (EDTA) .mwmu ' Hydrdxy ethyl eeiiuiose 05? 9.9 Phenexyethanei 0.7 {L7 Poiyethyiene giyeei 4th) Sodium hyalurenate Sodium hydroxide soiutien q. s. q. s. Eq.s. qE. s. topH Etepii EtepH Etepii .5 5.5 5.5 5.5 Purified water qs. q.s. q.s.

Phase A: Purified water, Edetate disodium dihydrate (BETA), Aliantein, Hydrexy ethyl. cellulose Phase B: d water, Sedium hyahrrenate Phase C: Phenexyethanei, Sodium hydroxide sohrtion Phase D: Giyeerin, Besifloxaein.HCi (Equivalent to Besifiexaein), d water, Sodium hydroxide sehatien Phase E: Poiyethyiene giyeei 400, Diethylene glyeoi meneethyi ether Phase F: Sodium hydrexide en re teem} Methed (Bf Preparetiert: 1) In a main mixing vessel, edetate disediurn and ein were disseived in water. Then hydrexyethyi eeihdese was added in n to the main mixing vessei while stirring at 1th“) rpm using over-heed stirrer. Geiiing agent was allowed. to swell at 100 rent for 30 min to get prooer hydration. 2) Hydrated onate sodium were added to main mixing vessel and mixed.

Then phenoxyethanol was added to the mixture. Then? pH of the mixture was raised to 6.0 with sodium ide solution.

U1 3) in a separate vessel, glycerin and hesitloxacin.HCl were dispersed with continuous mixing at 380 rpm for 10 min. 4) Bilute solution of sodium hydroxide was added drop-wise to the te vessel to adjust oil to 5.5.

) The contents of the above mixture were added to the main mixing vessel with ll} stirring at 260 rpm for 2 hours. 6) Finally, polyethylene glycol and diethyiene glycol monoethyl ether were added to the main mixing vessel and mixed for further 20 min. 7) White—to—pale yellow gel was obtained. {(93374} s: No viscosity drop was observed in hydroxyethyl cellulose to" based gels. Gels with hesifioxacin.HCl concentration upto 4% w/w (equivalent to hesifloxacin) were found to have acceptable consistency and sensorial characteristics. Results of ities of gel conioositions are given in Table 33.

Table 33: Viscosities of Gel Coninositions at Bifferent Concentrations of 29 Besii‘loxaeinfitfil Besifloxneinfifi‘l Sr. No. (Iornriosition equivalent to hesiflosaein Viscosity (m?a.s) (%w/w) 4 can 4 334:2. s Exemeie 37.}9reperation of Get Formniations Loetied with Bifferent Concentrations of Suspended Beeifloxecinfifi to e the Effect on Viscosities of Formniations Containing (Zerhomer Gel formuiations containing different concentrations of besifloxaein {It hydroehioride were formuiated as per the compositions shown in Tobie 34. These formuiations were ated with oerbomer to observe the effect of tration of hesifioxeeinHCE on Vieeosities.

Tabie 34: GeE Formeietions with Bifferent Concentrations of tfied Besifloxeeinflfl Using er for Comnositione Gig-14$ GLIS, GLlé anti Chemicai Name Comooeition (‘Vowz’oq BesifloxacinHCl (Equivalent to Besifloxacin) Diethyiene glycol monoethyl 5 555 ether Edetate disodium dehydrate (M m 0,1 (3,1 (EDTA) Giyeerin 5 5 Phenoxyethenoi (2.7 e7 e7 ‘ es: Poiyethylene glycol 400 5 2 5 we?0‘4 e4 (3,4 (3.4 godmdgmmqs to V (1,5. to es to q.s. to I : pH 5.5 Purified water q.s.

WO 14666 This page is blank.

The next page is page 178 Phase A: Purified water, Edetate disodium dehydrate (EDTA), Allantoin, Carhomer homopolymer type C Phase B; Purified water, Sodium hyaluronate Phase C: Phenoxyethanol, Sodium hydroxide solution Phase D: in, Besit‘loxaeinHCl Equivalent to Besifloxacin, Purified water, Sodium hydroxide solution Phase E: Polyethylene glycol 400, Diethylene glycol monoethyl ether Phase P : Sodium ide solution ll) {@3375} Method of Preparation: 1) in a main mixing , edetate disodium and allantoin were ved in water. Then carbomer homopolymer type C and hyaluronate sodium were added and allowed to swell at 2th rpm. for 60 minutes. Then phenoxyethanol was added to the carhomer mixture. Then, pH of the mixture was raised to 6.0 with sodium hydroxide on . 2) in a separate vessel, glycerin and hesitloxacin.HCl were sed with continuous mixing at 300 rpm for l0 min, 3) Dilute solution of sodium hydroxide was added drop-wise to the separate vessel to adjust pll to 5.5. 4) The contents of the above mixture were added to the main mixing vessel with stirring at 290 rpm for 2 it.

) Finally, polyethylene glycol and diethylene glycol monoethyl ether were added to the main mixing vessel and mixed for further 20 rnin, ) White-to—paie yellow gel was ed. {@376} Results: Addition of besiiloxacin.llCl to the gels has led to drop in viscosity in a concentration dependent fashion, although a minimum viscosity of about 3900 mPas can be considered acceptable. However, viscosities lower than 300% mPa.s would not be acceptable for desirable flow properties (from tube) and application on skin (by patients). Results of ities of gel compositions are 3t) given in Tattle 35‘ Tania 35: Visnnsities oi’ Gai Comnnsitinns With Biffereni Cnncentmtinns ni‘ Besifinxacinfiffii WWW—rm. i Besiflnxanin‘HCi (”Awlw) S.Nn. (inmpnsitinn equivaieni to hesifinxacin Viscnsity" {mPn.s) g (%wlw) 1 (31,14 0 i 5333 GLi 6 4 GM 7 in 1310 *measured by ietsr abQQ C~LTD SO/QC), Antnn Paar Exampie 38: Frennratinn ni‘ Gei Fnrmniatinns Containing Snsneniieii Nadifinxnein, i’rniifinxanin, Uiiflnxacin, BesifinxaninaHCi and Cnmhinatinns with Adnpniene; Frepaieai using Hyiirnxyetiiyi Ceiininse as Thickening Agent Ge] fonnuiations ning ded nadifioxacin, pmiii‘oxacini uiifloxacin and if) basiflaxanin were farmuiated alone and in their cembinatien with adapaiene as per the compositions Shawn in iabis 36 Thesa formuiations had pH of 5.5-6 and viscosity of around 4060 — 600i) mPa‘st i‘aiiin 36. Gei Farmniatinns Landed With Nadii’inxanin, P‘miii‘inxacin, Uiii‘inxacin and aiinn with Agingiaiene {Cnmnnsitinns GAL GAE, GAS, GAs’i and GAS) I Ciieniicai Name nmpnsitinn {% WM) Adapaiena besifiexacin néggéifimmwmm" Pruiifioxacin Ulifioxaein O (l Allantoin o a 0.2 9.2 Citric acid on (MS {3.15 l 0.l5 035 (315 Diethylene glycol monoethyl 5 5 5 5 5 1 1 J jether a Emmmwill ejiml“““ 0.1 TWA lGlyeerin 5 5 5 5 5 HroTsoE“““““W462 0.2 9.2 9.2 Illydroxy ethyl cellulose l .2 1.2 l2 1.2 Phenoxyethanol 0.7 l 0.7 9.7 0.7 0.7 Poloxamer 407 (l 0 O 0.2 {13.2 :Poiyuhysxggggsrzra““““““i“““““EW 5 W Pedal-hydroxide solution qs. {1.5. qs. .s.

Purified water gs. qs. l gs. qs. qs.

Phase A: Purified water, Edetate disodiurn dihydrate, Allantoin, Hydroxy ethyl Phase B: ed water, Hyaluronaie Sodium Phase C: Glycerin, Besilloxaein.HCl equivalent to besifioxacin, Nadifioxaein, Prulllloxaein, Ulifioxaoin, Purified water, Sodium hydroxide solution Phase l}: Purified water, Poloxarner 407, Adapelene Phase E: Polyethylene glycol 400, Diethylene glycol monoethyl ether Phase P: Phenoxyethanol lfl Phase G: Citric acid solution. Sodium hydroxide solution {@9377} Method of Preparation: Stepwise procedure to prepare gel is ned below- l) in a main mixing vessel. edetate disodiurn and allantoin were dissolved in l5 water. Then yethyl ose and hylauronate sodium were added and allowed to swell at 160 rpm for 60 min. 2) Then phenoxyethanol was added to the above mixture. Then, pH of the mixture was raised to 6.0 with sodium hydroxide solution. 3) In a te vessel, glycerin and hesifloxacinllCl were dispersed with continuous mixing at 300 rpm for it) minutes Dilute solution of sodium hydroxide was added drop—wise to the separate U: vessel to adjust pit to 55.

“J1 xx The contents of the above mixture were added to the main mixing vessel with. ng at 200 rpm for 2 h. 6) in a separate vessel, adapalene was dispersed in aqueous solution of poloxamer 467. This adapalene dispersion was erred to the main mixing ll) vessel. 7) Finally, polyethylene glycol and diethylene glycol monoethyl ether were added to the main mixing vessel and mixed for further 20 min. 8) White—to—pale yellow gel was obtained.

Example 39: ation of Gel ations Containing Combination of Suspended Besit‘loxaein.ilCl and Adapalene; ?repared using Carhomer as Rheology Modifier {@3373} Gel formulation containing suspended hesiiloxaein in combination with adapalene was prepared as per the compositions shown in Table 37. The formulation had pH of 5.5-6 and ity of around 4063 mPas at shear rate of . st e 37. Gel Formulations Loaded With Suspended Besitloxaein.HCl in Combination with Adapalene for Compositions GL1? liCheflfifital Name lEComposition (% wiw) GLESE BesifloxacinHCl equivalent to i j hesitloxaem ‘ Adapalene i). l WO 14666 er homopolymer type C Diethylerie glycol mouoethyl ether Edetate disodium dehydrate i..................______......................_........................... l Poloxamer 407 Polyethylene glycol 400 Sodium ide solution Purified water Phase A: Purified water, Edetate disodium dihydrate, Allahtoin, Carhomer homopolymer type C Phase l3: ed water, l-lyalurohate sodium Phase C: Sodium hydroxide solution Phase D: Glycerin, Besit’loxaeihHCl equivalent to hesiiloxaeio, d water, Sodium hydroxide solution Phase E: Purified water, Poloxamer 407, Adapalehe Phase P: Polyethylene glycol 408, Diethylerie glycol monoethyl ether Phase G: Phehoxyethariol Phase l-l: Sodium hydroxide solution toasts} Method of Preparation: i) In a main mixing vessel, edetate disodium arid allantoin were dissolved in water. Then earhomer and hylaurohate sodium were added and allowed to l5 swell at l00 rpm for 120 min. 2) Then phehoxyethahol was added to the above mixture. Then, pl-i of the mixture was raised to 6.0 with sodium hydroxide solution. 3) ln a separate vessel; glycerin and hesifloxacinllCl were dispersed with continuous mixing at 306 rpm for id mill. 4) Dilute solution of sodium hydroxide was added drop—wise to the separate vessei to adjust pH to 5.5.

The contents of the aheve mixture were added tn the main mixing vessei with stirring at 200 rpm fer 2 h. 6) in a separate , adapaiene was dispersed in aqueous soiution ef peiexamer 40?. This adapeiene dispersion was transferred to the main mixing vessel. 7) Finaiiy, polyethyiene giyedi and diethyiene giyedi meneethyi ether were added to the main mixing vessei and mixed for r 20 minutes. 8) White~te-paie yeiiew gel was obtained. i0 e iii}; Ge}. ations eentaining besifiexaein hydreehieride Ge] fermuiatiens eentaining hesifloxaein hydreehleride were prepared using earhemer as geiiing agent as per the eempesitiens shown in Tahie 38. The get formniations with acceptable Viscosities (3590-15909 mPa.s)and pH range (5.5 to 6.0) were obtained.

Tehie 38 Chemieai Name Cemnesitien (% wlw) {Eightiexaciniiei (Equivalent to * i 2 Besifiexaein) Edetate disediuin dehydrate genre) 0.1 ties Aiiantein e2 <15 Giyeerin 5 5 Carhepei 980 ene Giyeei Pelexamer 40’? i-Phenox-yethanei w - Peivethyiene giyeei 400.I WO 14666 Methyl parahen Propyl paraben Triethanolaniine ' Purified water Method of Preparation: 1) in main mixing vessel,edetate disodium, allantoin and poloxanier were dissolved in water, followed by addition of carhomer while stirring at 200 rpm using over—head stirrer. Gelling agent was allowed to swell at 290 rpm for 2h. 2) Phenoxyethanolor ns was/were added to the above mixture. Then, pH of the e was raised to 5.5 , 6.0 using triethanolamine solution. 3} lo a separate vessel, glycerin and hesifioxacin.llCl were dispersed with uous mixing at Sill) rpm for El) minutes. Dilute solution of ll) triethanolamine was added dropwwise to adjust pl—l to 5.5 to an. 4) The ts of the above mixture were added to the main mixing vessel with ng at 200 rpm for 2 hours, ) Finally, polyethylene glycol, propylene glycol and diethylene glycol nionoetliyl ether were added to the main mixing vessel at a stirring rate of l5 200mmfor further 26min. 6) if needed, pH for this mixture was further adjusted to 5,5 to 6.0 using anolarnine and the mixture is stirred for 2h to obtain white~to~pale yellow Example 4i: Gel formulations containing hesii’lexaein hydrochloride having hydroxyethyl cellulose and sodium hyaluronate as gelling agents Gel formulations containing besifloxaein hydrochloride were prepared using hydroxyethyl ose and sodium hyaluronate as gelling agents as per the compositions shown in Tahle 39. The gel formulations with aeoeptahle viscosities (350045090 miles) and pl-l range (55 to 7.0) were obtained.

Tahie 39 rCherhieai Name Composition (% wire) ' GLZZ GL23 €31,241 GLZS GL2?“ ] Besifloxaeirt.HC1 V ”MW—“fl aient to 1 4 2 Besifioxaein) I Eeths'iene giyeol monoethyi ether 5 5 Wm Edetate disodium dehydrate (EDTA) (1.1 (1.1 0‘1 1 0.1 (1.1 mm"mm ""5 5 5 5 5 i-fimi ose E (1.9 1.2 1.5 1.75 1.5 ”iiiiEECEiyahami or? 0.7 9.7 0.7 E a Polyethyiene gig/901491} 5. 5 5 5 Sodium hyaiuroaate 0.4 {1,2 (1 t) Sodium hydroxide so1ution qs. qs. ease (1.5. as 1Erit‘red water qs. qs. q.s. q.s.

Method of 13’reparation: 51) in main mixing vessei,edetate um was dissoived in water, foiiowed by addition of hydroxyethyi eeiiuiose(HEC) while stirring at 200 rpm using over— head stirrer° HEC was aiiowed to sweii at 200 rpm for 2h 2‘1 in a separate vessel, sodium hyaiuronate was aiiowed to sweil in water under stirring for 1. hr. After the compietion of sweiiing of both the thieitening , sweiied sodium hyaiuronate was added to the main mixing vessei, 3) Pheuoxyethanoi arid diethyiene giyeoi monoethyi ether were added to the above mixture. Then, p11 of the mixture was raised to 5.5 -= 6.0 using sodium hydroxide solution. 4) in a te vessei, giyeerin and hesifioxacin.HC1 were dispersed with 1.5 continuous mixing at 561} rpm for 20 minutes. Dilute soiution of sodium hydroxide was added drop~wise to adjust p11 to 5.5 to 6.0.

) The ts of the above mixture were added to the main mixing vessei with stirring at 200 rpm for 2 hours; 6) Finally, polyethylene glycol was added to the main mixing vessel at a stirring rate of 20971)me further 20min. 7) if needed, pH for this mixture was further adjusted to 5.5 to 7.9 using sodium hydroxidesoltition and the e is stirred for 2h to obtain white-to—pale yeliow gel. lixamoie 42: Qel formulations containing oesifioxaein hydrochloride with different combinations of eerhomer, hydroxyetliyl cellulose and sodiom liyaltironete as gelling agents Gel formulations eontaining besilloxaein hydrochloride were prepared using it) different combinations of er, liydroxyetliyl cellulose and sodium hyaluronate as gelling agents as per the compositions shown in Table 4i}.The gel formulations with acceptable Viscosities (350045000 m.Pa.s)and pH range (5.5 to 7.0) were ed. 1 5 Table 4% Mywe; e727: ems ‘ ems; ems Gen lBesifloxaeinHCl {Equivaent to flw—“fiww Besifloxaein) Ethylene glycol monoethyi ether 5 5 mgmmgw Edetate um dehydrate oi 0.1 0.1 W 2 a i a (EDTA) in 5 5 l-iydroxyethyi ose i.5 0.5 in 0.8 lCatbomei' 0.7 1.2 ()4 0,8 lPlienoxyethanol % 0.7 e7 97 0.7 3130lyethylene glycol 40o """ " ' i 5 5 5 5 "Soméjliiini-mctluronate is _o.2e {7.2 jiSodiumhydroxide solution 2 gas qs. qs. (7.5. gs.

Purified water L_______________.___________.____.__________._____~_______________.

Method at" Prenaration: 1) in main mixing vessel, edetate disodium was dissolved in water, ed by addition of carbomer and ,/ or hydroxyethyl ose (EEC) while stirring at 200 rpm using ead stirrer. Carbomer and HEC were allowed to swell at 200 rpm for 2h. in a separate . sodium hyaluronate was allowed to swell in water under stirring for i hr. After the completion of swelling of both the thickening agents. swelled sodium hyaluronate was added to the main mixing vessel. 3} Phenoxyethanol and diethylene glycol monoetliyl ether were added to the ll) above mixture. Then, pH of the mixture was raised to 5.5 .. 6.9 using sodium hydroxide solution. 4) in a separate vessel. glycerin and besitloxaoin.ll€l were dispersed with continuous mixing at Stilt) rpm for 20 s. Dilute solution of sodium hydroxide was added drop~wise to adjust pH to 5.5 to 6.6.

) The contents of the above mixture were added to the main mixing vessel with stirring at 290 rpm for 2 hours. 6‘) Finally, polyethylene glycol was added to the main mixing vessel at a stirring rate of 290mmfor further 20min. 7i if needed, pit for this mixture was further adjusted to 5.5 to 7.0 using sodium hydroxide solution and the mixture is stirred for 2h to obtain white-to-pale yellow gel.

Examnle 43: Stability Studies of Besil‘loxaeinj‘lCl Susnended Gel (GIL-15) llrepared Using Carhomer llllli‘ldll} oxacinHCl suspended gels were packed in laminated tubes and charged for stability studies under accelerated condition (49°Ci2°C, 75% RHfl‘l/B). These gels were ted for physical appearance? pH, viscosity, assay and content mity at initial time (T0), 2 weeks (Tm) and one month (Tim). ltlllildll Results: Results suggest that gel is stable under tested time durations.

Results are mentined below in Table 41.

Table 41. Evaiuatien et‘ stability samples el‘ besiflexaein.T-TCT gel (GLTS) put an accelerated stability eenditlens WWWWWWWESpeeilTeaimtigfl-"WWWMWW Tests E n lnitlaig’ili‘ (T2w) lmonth(Tgm) White to pale yellow; white gel of white gel 0f white gel of Description. Q hemegenenus uniform unifunn n : gel consistency censisteney consistency .. an . pH 5n 5.5 — 5e 5.5 — 5.9 5.5 ~ 6.0 ; Viseesity@ 25th— SGGG E 4322 mPas 42l2 mPas 4069 mPas °C 2G rpn1* mPas E E 0.95% — a Assay l.Gl% (2,9227% (2,5933% . E i.95% Centent l00.95% - 98.90% — 98.96% ~ 95% ~ 105% Unifmmity l0fi.44% 2 99.Q§% Phenexyethanc 0.63% - 9.71%. 368%, 6.69% E l Content 077% . E *measured by Viscometer (RheelabQC C-LTD SO/QC), Anton Paar Example 44: Stability Studies at” Besiflexaein.HCl Susnended Gel {GLTG} Prepared Using Hydrexyetliyl Cellulese } BesiflcxacinHCl suspended gels were packed in laminated tubes and charged fer stability studies under accelerated eenditien (400sz2‘3C3 75% l0 REE-{$5945}. These gels were evaluated for physical appearance, pH, viseesity, assay and centent uniformity at initial. time (To), 2 weeks (TN) and (me month (Tim). {@383} s: Results suggest that gel is stable under tested time durations, Results are mentined below in Table 42.

Tobie 42. Evaination of stehiiity sampies of hesifioxeeinfifji get {GLitB not on neeeierstett stahiiity conditions E’et‘iod of Stnhiiity Stony Tests E Specification E initial (T0) 2 weeks (TZW) 1 month (Tim) —i white gel of white get of white get of Description White to paie yeiiow, uniform uniform uniform homogeneous get consistency E consistency consistency E ' pit 5.0—6.0 555,0 77inscufonsity _ 531) a ' 2500—5090 rnPes 4028 mpafi 3674 mPes 3638 mf‘as °C , 26 item" Asst; o,9§éX-miiiis37ti 1.05% 134% inzinr’t Phenoxyethenoi , 1 0.63% — 0.77% 0.73% Content *measured by Viseometer (RheoiahQC, C—LTD Sim/QC; Anton Paar Exempie 45: Stahiiity Studies of Besit’ioxeein.f£€i Suspendett Cream (Ch/EM} t’repered Using Cathotner Etitt384} Besifloxaein‘HCi suspended creams were packed in laminated tubes and charged for stability studies under aeceietated condition (40°Ci20C, 75% ‘i/u). These creams were ted for ai appearance, pH, ity, i0 assay and content uniformity at initiai time (T9) and one month (Tim).

E69385} Resnits: Results suggest that get is stehie under tested time durations.

Resnits are. nientined heiow in Tahie 43.

Tahie 43. Eveiustion of ity samples of hesifioxaein.HCi cream {Ch/162) out on eeeeietated stehiiity eontiitions Petioti of Stehiiity Stndy E initiai (it) It month (Tmfl E mmm-i—mmm—fi E White to pate yeiiow, white white ption. E E homogeneous cream neous E homogeneous gViseosity @ 25°C a 201 2500 — .5000 ml’as 3084 ml’a.3 249:9 mPas Assay 0.05% - 1.05% 1.01% 0.987% yetlianol g 0.03% — 0.77% met/t 070% Content i 1 *measured by Viseometer (RheolabQC, Gill) SG/QC), Anton ?0ar Example 46: Stability Studies of Besit‘loxeein.ll€l Suspendetl Cream (CMllS) lireginretl Without Use of Polymer {$9386} Besifloxacin.ll€l suspended creams were peeked in laminated tubes and charged for stability studies under accelerated condition (400(32ng 75% ltllztSO/tp). These creams were evaluated for physical appearance, til-l, viscosity, assay and content uniformity at initial time (T0) and one month (Tim). $3387} Results: s suggest that gel is stable under tested time durations. it) Results are mentined below in Table 44, Tattle 44. Evelnetion ot‘ stability samples of besitloxaein.li€l cream (CMtlS) put on rateti ity eontiitions Period of Stability Study Tests Specification initial (To) 1 month (Tim) White to pale yeliowg White homogeneous white homogeneous; Description a h 3 3 ; eons cream 3 cream eream 1 pit 5.0 ~ 6.0 5.5 - 0.0 5-5.5 Viscosity @ 1506 ~ 35th mPas 1936 ml’as 1908 mPa.s %: 2t) rm* : Assay 0.95% — 1.95% (3.97% 0.967% Preservative Content 0.63% to 977% 0.75% 0.70% (Phenoxyethanol) *measured by Viscometer abQCg C—LTD Ell/QC).a Anton Paar Example 47: Stability Studies of Besii‘loxacin.ll€l Soluble Gel (SL7) ?repared Using yetbyl Cellulose {69333} xacinllCl suspended creams were packed in ted tubes and charged for ity studies under accelerated condition (40°Ci20C: 75% S RHeS‘Vu). These creams were evaluated for physical appearance, pl-l, viscosity, assay and content uniformity at initial time (Ta) and one month (Tim). {@389} Results: Results suggest that gel is stable under tested time durations. s are mentined below in Table 45. ll) Table 45. Evaluation of stability samples of besilloxacin.ll€l soluble gel (SL7) put an accelerated stability conditions ?eriod of Stability Study ’l‘ests Specification initial (T0) 2 weeks ('l‘gw) l month (Tim) Pale yellow j Pale yellow Pale yellow Pale yellow ption. ‘ transparent gel transparent gel transparent gel transparent gel pli 4.0 ~ 5.0 4.5 - 5.0 4.5 — St) 4.5 ity {it} 25°C, l 2506 — 506i) ml’as 3045 ml’as 266i mPa.s 2700 mPa.s 2t) rpnr* Assay 0.95% .. 1.05% "0.99% m 7.00% 0.99% Pbenoxyetlianol 0.63% — 0.77% (),7l% 0.73% (3.71% Content graces-uteri lay-Visconietei (RbeolabQC, C—E’l‘lll SG/QC), Anton Paar Example 48: lleterrnination of tire antibacterial efficacy against antibiotic— nonrespotler P. series of besit‘loxacin (APl) by time kill experiment. lililSlltil Procedure: P. genes (CCARM 9610) aqueous suspension (G5 McFarland standard equivalent) was centrifuged at 2900 rpm for 20 min, pellet was resuspended in Brain Heart infusion (Bill) broth. Resultant P. genes suspension was kept for overnight (l6 h) incubation in anaerobic box at 37°C.

Stock solution of oxacin.l-lCl (l trig/ml) was prepared in dimethyl sulphoside (DMSQ) which was further diluted with Bill broth to achieve g stock of besilloxacinillCl (25 rig/ml). 'l‘hen reaction mixture was prepared by adding 906 ul of P. atoms 05 McFarland rd culture (after l6 h incubation) to lth gal of besitloxacinl-lCl working stock solution (25 rig/ml), final besiiloxaein.HCl concentration in reaction mixture was 2.5 gig/ml. Reaction mixture ( l ml) was incubated at 379C. for 24 h in an anaerobic box in two sets» (l) tube with P. genes with 2.5 ug/ml BesifloxacinllCl and 2) broth control without besilloxaein.l-l€l. At predetermined time points (i h, 6 h and 24 h), cells were ted and plated in duplicate after serial dilution in Brain Heart 'lniilsion agar l0 plate Plates were allowed to grow for 3 days at 37°C in anaerobic box. After incubation.) plates were counted to determine colony g units (CPU) and log reduction was calculated. in a similar experimental set—up, water was used as solvent d of DMZ-St} and effects of solvents on time lzill cs were also studied. l5 {96391} Results: BesiiloxaeinllCl sample (prepared as l il stock in water and DMSO) showed similar time kill kinetics against P. genes (CCARM 96m) at all the tested time point (Table 45 and Figure 12) Table as. Time kill cs of Besit‘losacin.ll€l against P. series, studied at concentration of 2.5 2; ml at l h, a h, and 24 ll. '"l‘irne (h) Besi‘loxaeinllCl in Besilloxaein.ll€l in Blah Cflfllml .5ftnl a___nrtisomi 0 7n a are 7.0 a ens 7.0 a 0.02 1 l 7.2 a one __________ 6 a 7.4 a no"; i 2.4 5.2 a are t “5.2 a one as _+_ on Example 49: Antimicrohial susceptibility of P. arenas isolated from acne patients (clinical isolates) {($392} The antimicrobial susceptibility of P. acnes isolates against various antibiotics was determined by micro broth dilution method as follows- WO 14666 {@9393} Procedure: P. genes were culturedin Brain l-leart infusion Agar (BHlA) at 370C for 48 hours under bic condition. For MlC test, Bl—ll broth (lOO til) was added into all 96 wells and liltl ill of broth containing drug was added to first well (lA to ill) and serial (double) dilution was carried out for up to U1 it) wells (column l to column ll) of 96 well plate). For ial inoculurn, P. acnes culture turbidity was adjusted to t).5ilvlcl1‘arland stande (approximately l5 it it'll) and r diluted (109 times with sterile Elli broth). d P. acnes suspension (ltll) ill) was added to each well except sterility control wells (column l2 or" 96 well plate). inoculated nlates were incubated at 37°C for 72 hours under it) anaerobic condition.Afier incubation, MlC was determined by adding Alamar blue dye. lllllllllall Results: The MlC results on P. acues clinical isolates susceptible to clindamycin and erythromycin) indicate that all the strains arnycin and erythromycin susceptible) are susceptible to rnycin, erythrornycin, minocycline and besilloxacin (Table 47), interestingly, wide variation in ivity to clindamycin and erythromycin were observed in Clindamycin— nonrespondee clinicalP’. acnes isolatesAll of the antibiotic—nonresponder clinical isolates were found susceptible to besifloxacintt'l‘able 48), Table: 47. P. series clinical isolates {susceptible to cllndamycln and erythromycin) antimicrobial suscentiblllty against clindamycin, e tbrorn 'cln, mlnocveline and besifloxacln P: defies a _ Actives Mitt: strain No. v3.9 <a.a"3 «on; one 0.25 ‘ vs—lo «are 43.03 0.25 we are <03 — 0.25 _ a aaaaaa V4~8 43.03 arcs (>25 V4-13 «are <ae3 0.06 9.25 v5-2 «are «2.03 see as ‘ i 4103 0‘5 <0.tl3 ace . as V66 <O.(l3 <0.03 one 0,5 V05 <0.03 <00}. 0.05 0.5 V6-7 <0.03 <0.03 0.05 0.5 i V723 <0.03 <0.03 0.05 0.5 .. .. * 1 V74 «0.05 <0.03 0 0.5 51714va ______ <0.03 <0.05 _ 0.051%“ 0.5 * V9512 <0.15 <0,09 - 0,5 712-25: 0.25 5113—37: 0.5 0 <015 <0.09 _ 7 05 1.775777. <00 - 5.25 75737 <70 7.7 V184E _______ __ ““3013 M050 wm 0.25 VVVVVVVVVV MM V1020 $.15” 30:00“ .. 0.25 __w _____ _ J VZG—SE {0.15 <000 — 0.25 1251110: 48. P. acnes 011111001 150Eates {01513113111315.2111 resistance) antimicmbiaE susaeptibifity agaimt 01indamycin, erythmmycén. tetracyefine 311d 11051110313010 A011V0§ MEG 7195 0:07:05 . 101113010_ Erythmmycm, Tetracyeime, 15031110150011}. . strain N0. 91%”m1) W_____ (5/““1 mww V21A-1 1—8 0 (1.25 0.25 V21A—2 8—64 50.200 0.25 a 0.25 W m ------Tm...

V21A—4 100200 . 025 V21A—5 1111)2(10 (125 1 VZIA6 1-2 E.........Y.21A“7 WWQWSWZWWW 100—200 (1. 25 (1 13 V21A~_8 ”1:2100200 {125 (1.25 3 V21A-9m16—32101)290 . 0.25 0.25 “““___:W 100200 - : ......... __9.2§.__W >209 0.5 VZIBWJW________..__,W_3%99___W_.__W?;§W__________.

V2105 15—754 >200 05 V21 15-7 {@395} Conclusion: The lvfllC results presented in Tables 45-46 shows that P. aenes isolated from volunteers V3 and VZl were resistance to clindamyein and omycin, but were tible to hesitloxacin.

Example St}: Comparative in vivo aeoltinetie profile of Bifferent Gel and cream formulations of Besifloxaeint {eases} Fharmacokinetic (PK) profile of a topical anti-microbial formulation is important from two perspectives, Firstly, it ines whether the ation l0 can deliver above MlC level concentration of antimicrobial agents to kill the pathogen at the relevant layers of skin for a prolong period of time. Secondly; PK study determines whether penetration of the s into systemic circulation. has crossed the allowable limits: Formulation with better retention in the skin layers and low penetration in blood would be ideal. l5 {693W} Method: 8-10 weeks old Sprague Dawley rats were randomized into three groups according to their body weights. Fully suspended l% Besiiloxacin gel (VLN~Fltil/BSF/GLIOtSS), fully soluble l% Besiiloxacin gel (VLN~ F2l/BSF/GL/OlllA) and fully ded l% lr'lesifloxacin gel (VLN- FZG/‘BSF/Clb’lltldr) were used for comparison purpose. Animals (4 per time 2% point/formulations} were treated with topical applications of test formulations as a single dermal dose of 50 mg / 25 cm2 (hair was clipped from 5x5 cm area in dorsal flank of the animal. a day before application). After application 2 minutes were allowed for drying. The application area were then covered with a non— absorbent surgical tape (TegadermTM) Blood was collected from retro-orbital plexus at preudose (G h), 0.25, 9.5, l, 2, 4, 8, 12 and 24 h postwdose. At the end of each time point, animals had been euthanized and "legarlerrnm were removed and ted. Then the d area was gently wiped with water—dipped cotton balls to extract drug efficiently from the top layer or Stratum Corrzeum. Finally? applied area of the skin was excised; With established extraction procedure. Besifloxacin 3C was extracted from the shin samples. The skin and plasma s were analyzed by LC~MS/MS to know the concentrations in each matrix and obtained data were used to caieuiate Cmaxt Tmaxr ti/25 AUC. The results of PK study using different formulations Besirioxaein gel were plotted in Figure 13. {933%} Resuits: ative PK data showed that both fuiiy suspended and U1 fuiiy soiuhie Besifioxaein get i‘onnuiations have the abiiity to deliver and retain higher than MiC concentration of Besifioxacin in the shin even after 24 hours of appiieation, Their Crnax vaiue is same as the Mutant Prevention Concentration (MPC) of Besifioxaein against P. acnes. Though fiiiiy soiuhle one had exhibited higher retention profiie in the skin than fiiiiy suspended ones, yet aii of them i0 showed iow penetration in niasrna (below detection iimit). These data suggest that aii three formulations are of sustained release in nature . Therefore unique formuiaries deveioped here oouid penetrate at the site of infection in sufficient . quantity t endangering the host safety.

Exanipie Si: ieai evaiuation {minimum inhibitory eoneentration9 zone of inhibition assay and time itiii assay) of Besii‘ioxaeindiCiforrnuiations against P. aenes MTCC i951 (snseeptihie strain) {iiiifidi Antibacteriai activity of hesifioxaein formuiations were tested against P. genes MTCC 119951 by various antimicrobiai susceptihiiity methods. The foiiowing sampies were anaiyzed: Tahie 49. Besii‘ioxaein.§i€i ations tested t P. eerres MTCC i951 Formuiation Betaiis Besii‘i.oxaein.ii€i Content % w/w i ,BesitioxaeinHCi i [ Suspended Cream {1% a 9C 136 BesiiioxaeiniiCi e 2 Git/[0&3, “7 i Cream (1% w/w) a ___.....A........_A__ BesifloxaoinHCi (EL/Q20 1.i2 Suspended Crei (1% w/w) ititidtitii Procedures n P; acnes were caitureo’ in Brain Heart infiision Agar (SHEA) at 37°C for 43 hours under anaerobic condition. i.) Minimum inhibitory Concentration {MET}: For Mic test, Blil broth (100 all was added into all 96 wells and lOll ul of broth containing drug was added to first well (lA to ill).

Serial (double) dilution was carried out for up to l0 wells (column l to column ll) of 96 well plate).

L1] {titlaitlll For bacterial inoculunis, P. aortas culture ity was adjusted with 0.5 McFarland standard (approximately id x int) and further diluted tolutl times with sterile Bill broth. Diluted P. aortas suspension {ltlll iii) was added to each well except sterility control wells (column l2 of 96 well plate). inoculated plates were incubated at 37°C for 72 hours under anaerobic conditionsAfter incubation, MlC was determined by adding Alamar blue dye. 2) Zone of inhibition (Zfii): For ZOl test, Bl-lA plates were spread with ltltl ul of P. genes suspension (0.5 McFarland standard equal). Test samples (drugs/formulations) were dissolved in water/solvent based on the solubility. Sterile disc (6 mm) were loaded with it) ul of test samples (of various tration of drug), and were placed above the l5 plates containing P. acnes e. Then, plates were incubated at 379C for 48 h, followed by their ZOl measurements. 3}Tinie Kill Kinetics (TK): For TK Testf. genes (CCARM 90l0) aqueous sion (9.5 McFarland standard equivalent) was centrifuged at 2003 rpm for 20 min, pellet was resuspended in brain heart infusion (Bill) broth. Resultant P. bones suspension was kept for ght (l6 h) incubation in anaerobic box at 37°C. Stock solution of besitloxacinl—lCl (l rug/nil) was prepared in dimethyl sulphoxide (DMSO) which was further diluted with Bill broth to obtain working stock of besifloxacin.l-lCl (25 rig/ml). Then reaction e was prepared by adding 900 ul of P. acnes (3.5 McFarland standard equal culture (after l6 hof incubation) and lllfi ul of besifloxacinllCl working stock (25 ,ug/rnl), to obtain final besifloxaeinllCl concentration in reaction e, 2.5 gig/nil. Reaction mixture (1 ml} was incubated at 379C in an bic box- (l) tube with P. genes and 2.5 ug/rnl Besifloxacin.liCl and 2) broth l without besitloxacin.llCl for 24 h. At predetermined time points (0 h, 2 h, 8 h and 24 h), cells were ted and plated in duplicate after serial 3t} dilution in brain heart infusion agar plate. Plates were allowed to grow for 3 days at 376C in anaerobic box. After incubation, piates were counted to ine ceicny farming units (CPU) and icg reduction was caicuiated, EtifldiiZi Resuits: Concentraticn-efiicacywtimecurves (iabie fig-Si}, Fig. iii) indicate besiflexacintHCi gel and cream formulations (Tahiti 49) can have L1: differential antibacteriai activity against P. acnes, suggesting that ferrnulatiens can meduiate the iinai anti~acne efficacy of an active agent. For the various besificxacin formuiatiens tested, the MIC values were in the range of (LB-0.25 uglrni against P. genes MTCC i951 {Tabie Sit} Zone of tion assay results dispiay that aid besifiexacin formuietions (cream and gei) have antibacteriai 16 activity against P. acnes MTCC 195i. Ameng the three different tested ionnuietions, ZOE was found to be better with (EL/020 (Tabie 51).”?irne kiii kinetics s te thataii three formulations (GL/GZG, CR/Ofili, (IR/029C) showed r activity against P. acnes MTCC i951 at aii the time point. in addition, dose dependent differences in iciii kinetics were observed in aii three femtuiations between xacin.HCi concentrations of i rig/mi and 10 rig/mi.

{Tahie 52 and Figure iii) Tabie 5%. Minimum initibitnry concentratien (MEG) MEC et’ Besii’ioxnein formuiations against P. ecrres MTCC 335}. we; Ne Cit/2st: Cit/ens err/2e ? 1 4 4 2 2 3 . r "fame 51. Zane of 1n111191t19n (Z111) 91° besiflnxacin.11€1 gei and cream fnrniaEati9ns against P. genes Besifinxacin Creams 261 aainst R, genes MTCE 1951 ________.—mmmu.,___________ Average: 1191191 "1‘a1119 52. Tints: K111 Kinetics (1K) 91 besifiaxafiniifl g91 and cream fnrrna1at19ns against P. games 191TC€ 1%51 Effect 91‘ F9rnrnEat19n Treatment an E 997295 MTCC 1.?51 €911 Cnnnt (L9g111 9 (3111111111 {11} GLIQZfi CR111113 {3117112913 Bmtn 11 m1 111 ;~'rn11 1 111 1 14;; m1 111 2:;1Vl1 C9ntr91 L9 7.94 17.94 7.94 7.94 7.94 7.94 1 _______ _______7.94 2 7.79_________ 7.49 7.75 7.48 7.76 7.49 7.99 9 7.49 6.9:: 7.44 7.91 7.43 6.98 9.34 * 24 6.94 5.44 6.86 5.49 6.83 24 9.92 9.98 9.93 9.94 9.92 9.96 9.95 ExampEe 52: ica1 Eva1nat19n {minimn- innihittary 99ncentratien, 29119 91" 11111113111911 assay anti time 16111 assay) 91 119si119aa91n.11€1 (A111 9941 fnrma1ati9ns) against S. aureus MTCC 911118 anti F. 991293 N11”CC 1951 113} AntibacteriaE ty Of besiflnxaein APi anti basifioxacin containing fonnuiation against S. aureus MTCC 69118 wcre datcrrnined. The following s9mp1es were used far tha study.

Tania 53. Exeniniary ations for determining the antibacteriai activity against S. acreage M’i‘fiC d9ii$ S. New s Contents 1 BZMnl Unmodified besifloxacin 2 Stearic acid coated Unmodified BSF ________ WfigMWIZW W WW3 BZM—3 Stead id coated modified BSF 4 BZM—4 Stearic acid coated modified ESP a BZM—S Stearic acid coated modiied BSF 6 BZM—d o ei 7 _ 8 ‘ 9 BSD-8 SA coated BSF NPS Unmodified BSF Steaxic acid coated Unmodified BSF Unmodified xacin Lauric Acid coated modified BSF LA coated BSF NPS ’i‘anie 54. Exempiary forniniaticns for determining the antihaeteriai activity againei S. aureas MTCC 6968 Contenta W Saninieutjode _ BTK—i Unmodified Besifloxacin BTK-Z Stearic acid coated unmodified beeifloxacin Stearic acid coatedodifiedbesifioxacin Stearic acid coated modified besifioxacin Stearic acid coated modified besifioxacin ETK-é Placeboeei {£39494} nre: (i) m inhibitory concentration (MiC): S. aureus MTCC 6908 were grown in Tryptone Soya Agar (TSA) at 37°C for 24 hours. in 96 well platea 100 iii of Tryptonc Soya broth (TSB) was added into ail weiis and then EGG id of broth containing drug was added to first weii (1A to iii).

Serial (double) dilution was carried out for up to if) wells (column l to column ill of 96 well plate). S. aureus MTCC 6908 culture turbidity was adjusted to (3.5 McFarland standard (approximately l5 s ills) and firrther lllll times diluted with sterile TSA broth. S. aureus MTCC 6908 suspension (illil ul) was added to each well except sterility control. Plates were incubated at 37°C for 24 fter incubation, MlC was determined by adding Alarnar blue dye° {2) Zone of inhibition : S. nureus MTCC 6963 were grown in ne Soya Agar (TSA) at 373C for 24 hours BHA plates are spreadcd with lllll ul of ()5 McFarland standard equal bacterial suspension. Test samples (drugs/formulations) l0 were dissolved in water/solvent based on the solubility. Sterile disc (6 mm) were loaded with it) ul of test samples (of various concentration of drug), and were placed above the spreaded plates, Plates were incubated at 37 C for 24 h, followed by ZOl measurement. {3) Time Kill Assay: 05 McFarland standard equal S, oureus culture was prepared in sterile water. Besifloxacin gels were diluted in Tryptone Soya broth (TSB) to get final concentration 01525 gag/nil° Then 900 gal of S: auraus 0.5 and standard equal culture and ltlll pl of diluted Besifloxacin gel was mixed to obtain final. Besifloxacin concentration, 2.5 tag/ml.

Total reaction mixture i l ml) was incubated at 37°C in a tube rotator. After 2 and 6 h exposure, bacterial suspension was placed on Tryptone soya agar plate and incubated at 378C for 24 h. {@0495} Results are shown in Tables 55-56 and Figure 11. As show in Table 55, MlC results show that all the formulations (BZMl—BZMS dc; BGBl—BGBo) were similar in MlC except the o gel (BZM 6). Further, Laurie acid coated BSF (BSDl-BSDS) and fied BSF (ESE-9) Al’ls showed similar efficacy in l‘vllC assay, but stearic acid coated Al?’ls (BSBdBSDlfl) had less antibacterial efficacy. {994%} As shown in Table 56, zone of inhibition (ZOl) results indicate that all the ations BZMS) had similar MlC except the placebo gel (BZM 6), There was no difference between unmodified besitloxacin or Stearic acid (SA) coated besifloxacin APl based gel. l-lowever, stearic acid coated APl (BSD-6, WO 14666 BSD—7, BSD-8 and BSD-10) dispersion showed less or no antibacterial efficacy with respect to the unmodified BEEF A‘Pi (BSD 9) dispersion, Etifidti’i} As shown in Tabie 57 and Figure 1L the time kiii resuits te formuiations containing unmodified BSF or modified BSF have similar {J1 antihacteriai kiii efficacy Piacebo and growth eontroi showed no undesirable inhibitory growth pattenns.

"Faith: 55: MiC resnits Besifioxaein APE/Formniations MEC t P genes MTCC i951 E i—mwm—Tm-mmm—MWWMW .

Formniations E APE Bispersion fiuamafimem VVEE Gaga WEv EEE Eugfifimam .3 wEEHVa maVN "gm EQEQQSMM 93$. mgflfiafiafi 34% ma aa w g EEEEEMM gmuggfiwgm SE V E MQN EEE M MN:MN g m E? VSEEam 55.3... TENm Tabie 57: Time kiii Besit’iexaein AFi/ Fermniatiens Time Kiii aainst S aureus MTCC 6988 S. i initiai ZHrs Leg ZHrs Leg Ne. Samie {Terie Leifi Leiti Change (Leiii Change Grewth Centrei 6.66 800 1.94 -2.33 Examine 53. Minimai inhibitery Ceneentratien tieterrninatien fer Besit‘iexaeinfiCi Leatieri Fermuiatiens (Seini’iie Besifiexaeinifitfii flei, S Snsneniieei iexaein.ii€i Geis anti Snspentieti BesifiexaeinJ—ECE Cream) Against iiifferent Strains {if P. acnes {MTCC if‘iSi ntibie strain), {SCARM 88H} (resistant strainfl 388488} m inhibitory concentrations of gel and cream atiens containing besiflexacin.HCi against twe strains of P. genes {MTCC i951 (susceptible strain)? CCARM 9610 (resistant strain)} were determined. The feilowing s were used for the study.

Tabie 58. Exemniary fermniatiens fer rieterniining the Minimum inhibitery Ceneentratian (MEC) against P. ewes EMTCC i951 (stiseeptibie strain), CCARM 981i} {resistant strainfl Cede fietaiis Fermuiation is +..__._.___ ‘V’LN- * Besifiexacin.HCi seiubiizeci gei F_2 i/BSF/GL/GG i A VLN- Besifioxacingi-ECE suspended gei PEI/BSF/GL/OtifiA ' H 'drex etii E ceiiuiese ‘ BesifloxacinHCi suspended gei with carbemer hemepeiymer ‘ F3 9/BSF’GL/O‘ii A VLN- BesifiexaeinHCi cream with 4 CMGZ i F20iBSF/CR/003A HOmQ Oi 'mer T e C : CMOS VLN— BesifloxaeinHCi cream without V" F/CR/QGIEA ' Home 0! met TV e C ____________ _ illii4tl9l Procedure for Minimum inhibitory concentration tMiC): P. genes were edin Brain Heart infusion Agar (Bl-lift) at 37°C for 48 hours under anaerobic condition. For MIC tests Bill brotb if lGO ul) was added into all 96 wells and lGO ul of broth containing drug was added to first well {lA to ll—l) and-serial U1 (double) dilution was carried out for up to it) wells {column l to colunui ll) of ‘96 well . For bacterial inoculum, P. genes culture turbidity was ed to 0.5 McFarland standard {approximately l5 x 108) and further diluted (100 times with sterile Bill broth). Diluted P. antes suspension (130 ill) was added to each well except sterility control. wells (column. l2 of 96 well plate). inoculated plates were l0 incubated at 37°C for ’72 hours under bic conditionAiter incubation, MlC was determined by adding Alamar blue dye. llitiéiili Results: All besitloxacin formulations had shown similar MiC values 0.25 pig/ml). Results are shown in Table 5?. l5 ’l‘able 59: Results of Wilt? of gel and cream formulations against E acnes EMTCC 1951 (susceptible strain), CCARM sine (resistant strainfl MIC Value Against iiii‘t'eient P. series Formulation Cede Details strains? : 1 Codes i ' P. £65383 MTCC P. noises CSARM l9§l 9iilll SL7 tinuwtiiiunmeust/GL/smn iiiiiiiii GLE 5 i-ELN-F21/BSFIGL/002A : -ii t wes l A 2 E VLu—tsausnca'ecsn CMOS Vin—moress/cn/esztn Example 54: Minimum inhibitory {Zoncentratiun (MEC) Beterminatiou of Gel Formulations Loaded with Besifloxaciniiitfl i Adanalene / Combination against E acnes Eli/ETCC 1951 (susceptible strain), C€ARM §tlltl {elindamycinnonresponderstrainll {illisllli Minimum inhibitory concentrations of gel formulations containing besitloxacinilCl or adapalene or their combination against P. genes [MTCC l95l (susceptihie h CCARM gfiid (resistant strain)} were determined The foilewing sampies were used for the study, Tahie 6i}. Exemnieiy fermniatiens for determining the Minimum inhibitery Ceneentratien (MiC) against P. genes {MTCC i951 (enseeetihie )g CCARM 99H} {resistant strainfl Farmuiam“ Cede Detaiis (remains Cedes t g t {3ng BesiflexaeinHCi Q/BSF- nded) and Adapaiene ADP/GL/OGZ nded) Gei with t__ E hvdrex eth ii eeiiniese (31,19 Besiflexaein.HCi VLN—Flgz’BSF- 2 (suspended) and Adapaiene ABP/GL/OO3 (suspended) Gei with i heme eiymer iv e C BesitiexaeinHCi {soiubie} VLN~F 1 7/881:— and ene (suspended) ADP/GL/001 t with h drox 'ethvi eeiiuiese t SL7 VLN~ BesifioxaeinHCi soiuhiized 1:2 i/BSFIGL/GGiA ei i ADGi Get eentaining suspended VLN~Fi 9/ADP/GL/001 e Adaaiene {@412} i’reeednre fer Minimum inhibitei‘y eeneentratien (MEG): P. (zones were eulturedin Brain Heart infusion Agar (BHIA) at 37°C for 48 hours under it) anaerobic ennditien. Fer MiC test, Biii breth (100 ui) was added inte aii d6 weiis and 1% ui of broth containing drug was added to first weii (EA to Hi) and seriai (deubie) diiution was carried out for up in 10 weiis (eoiumn i te column 10 of 96 weii plate). Fer bacteriai ineeulum, P. genes euiture turbidity was adjusted to 0.5 McFarland standard {approximateiy 1.5 x 108) and r diiuted (106 times with steriie EH1 hreth). Diiuted P. acnes suspension (100 ui) was added te eaeh weii except steriiity eontrei weiis (coiumn 12 of 96 weii piate). Inoculated piates were incubated at 379C for 72 home under anaerebie eondition.After incubation, MiC was determined by adding Aiamar blue dye. {flit-”113} Results: All the ations except gei containing adapalene alone had shown similar MIC (0°13 pig/nil) against P. acnes [MTCC 1951 (susceptible strain)? CCARM 901i) {resistant strain) Results are shown in Table 61.

(I! Table 61: Resnits at” MEC of get forrnniations against 1? nenes ii‘vi'i‘t’jC 1951 (snseetible strain , CCARM 1:010 (resistant strain M11? Value Against Different R its. Formulation {genes strains (gig/nil} ‘ €ode Betaiis a No. 531195 R names ' R aenes ‘ refine 1051 SCARM 901.0 i {37ng Vina?rainsrnor/oir/002 0.13 0.13 2 (11,19 Vi...N-F1g/BSF—ADP/GL/t103 0.13 0.13 3 s30: VLN-rinner-Aon’oL/(iei1 0.13 0.13 4 SL7 Vin—r2i/asr/GL/001A 013 0.13 311 ____ja:a_?aeagiiiii >4 >4 Examnie 55: Minimum inhibitory Concentration (it/11C) Determination of Get and Cream Forrnniations Loaded With Besifioxaeinflt’ji against S. entrees MTCC 6968 {$6414} Minimum inhibitory concentrations of gel and cream formulations containing besi.tioxaein,HCl against S. aureus MTCC 6908were determined. The samples used for the study were same as those mentioned in Table 58.

} Froeednre for Minimum inhibitory concentration (MKS): S. aureuswere culturedin Brain Heart on Agar (BHIA) at 3?°C for 24 hours under anaerobic condition. For MIC test, Bill. broth (100 nl) was added into all 96 wells and 100 gal of broth containing drug was added to first well (1A to 11-1) and serial e) dilution was carried out for up to 10 wells (column 1 to column it) of 96 well plate). For bacterial um, S. aureuseuiture turbidity was adjusted to 0.5 and standard {approximately 1.5 x 108 ) and iiirtlier d {100 tirnes with sterile 31-11 broth). Eiluted S. aureussuspension (108 iii) was added to each well except sterility eontrol wells {column 12 of 96 well piate) WO 14666 inoenia‘ted plates were incubated at 37°C for 24 hoursAftet tion, MEC was determined by adding Aiamat bide dye.

} Resnits: Aii the fonnniations had shown simiiax MIC (0.25 gig/mi) against S. aurens MTCC 6908. Results are shown in Table 62.

Tanie 62: s ni‘ Witt: (if gei and cream fermnintinns against S. nnrene MTCC nine MIC VaineAgainst S.

Sr' Fernin a wi g“ n Cede Detaiis nnrens MTCE‘ $938 Ne. (Aides V ,2 mi) 1 SL7 VLN-F2 i/BSF/GL/QGIA (3.25 2 VLN—FZt/BSF/GLIOGZA 0.25 3 new VLN—Fi9/‘BSF/GL/051A 4 ennz VLN—FZO/BSF/CRJOOEA CM05 VLN~FZWBSFICRJGG4A 0.25 Exampie 56: Minimum inhibitnry Cnneenti’ntinn (MEC) Determinatinn of i0 Gei Fermniatiens Leaded Witt: Besifiexaein.H€i I Adapaiene /' Cembinatien of both Against St nurens MTCC 59%}8 {£53417} Minimum inhibitory eeneentratiens 0f gei feminintinng containing xneini—iCi er ndnpniene or their eombinatinn against S. aureus MTCC 6908wete determined. The sainpies used for the study were same as these given in i5 Tania: 6e. {$9413} Preeedni‘e fer i‘viinimntn inninitnry enneentt‘ntien (MEC S. aureuswete euiniredin Brain Heart Infusion Agar (BHIA) at 37°C for 24 h under anaerobic condition. For MIC test, BHI broth (190 iii) was added into aii 96 weiis and 100 iii of ninth containing drug was added tn first weil (1A tn 1H) and seriai 28 (double) dilution was carried out for up to 10 weiis (column 1 to coitimn it) nf 96 weii plate). Fer bacteriai inoeniutn, S. aureus enitnte turbidity was adjusted tn 0.5 McFarland standard (approximateiy LS x. 103) and further diiuted (166 times with e BHI broth). Diluted S aureussuspension (10% ni) was added to each welt except ity eontroi wells (column 12 of 96 weii plate), innenia’ted piates were ineubaied a: 370C for 24 fier incubation, Mic was determined by adding Aismar biue dyed {($419} Resuits: ARE the femulalidns except gel eeniaining edepaiene aieue had shown similar Mic. (0.25 gig/mi) t S. aureus MTCC 6908. Results are shown in Tahie 63.

Enable 63: Resuits sf MEC 0f ei fermuiaiiens seinst S. merges MTCC 69338 i MK: Veiue Against Fermuiatieu S. N0. ' Cede Betaiis S. guesses MTCC i a Codes ‘ mi 3 GL1 8 VLN-Fi9/BSF~ADF/GL/Ci)2 0.25 Uh-vthN GLig VLN-F19/BSF—AE‘P/GL/{3G3 (3.25 8530i VLN-Fl 7/BSF-ADF/GL/OOE 0.25 VLN—FZi/BSF/GL/OG i A vm—Fis/Aup/Gmm e 57: In vitro Time—kid c Study ei‘ BesifiexaciuflCi Leaded GeE and Cream Fermuietiens Against Stafiiiyiawceus diuresis MTCC 5938 EfifldZfi} In vitreTime kiii kinetic study of gei and cream fermuialidns eentainiug besifiexacinHCi against S: aureus MTCC 6968 was performed. The foiiowing samples were used fer the study.

Table 64. Exemgflary fermuiatieus (Gei and Cream} far In vim; Time—KER Kinetic Study against S. aureus 1%?ij 69% Fermuiaiiee Cede Betaiis Centents {jade VLN- BesifiexaeinHCi soiublized F2E/BSF/GL/GOIA ei i VLN— BesiflexacinHCE suspended FZi/BSF/GL/GOZA ei v eih'i ceiiuidse xacinHCi suspended gel with Carbomer F1 9/’BSF/GL/GS 1A ’ Hemepoiymer Type C, ““““““““““WW l VLN” e 4 cairn Ca? 0b inat” H Em smear/camera 01310903 Ct 'l.‘ e C oxacinl—lCl cream Uh without Carbomer F20/BSF/CR/GO4A Homcelymer Tye C Placebo gel Fl9/BSF/GL/(l70-P FZfiI’BSF/CR’GQ-l—P {93423 Preeedure for {rt-vine Time-Kill Assay: Besiflcxacin formulation was diluted in water to prepare l trig/ml steak and further diluted in Bill broth to make final eoneentratien of 100 rig/ml. S. aureus MTCC 6908 (900 ill) (3.5 {)1 McFarland rd equal e and MG ill of d Besifloxacin (llll) gig/ml) were mixed to cbtain finale Besillexacin tratien ll) ille in reaction mixture. Total on mixture (l. ml) was incubated at 37°C in an incubator. Pest incubation at l, 3 and 6 h, cells were plated after serial dilution in brain heart infusion agar plate. One tube with two read outs were taken for each sample. Cells l6 were allowed to grew for l6«24 h at 37°C in ineubatcri {lltlétZZl Results: All Besifloxacin formulations except Placebo had shown similar antibacterial effieacy against S. aureus MTCC 696% (Approx 2 leg reduction at 6 h). Results are slrcwn in Table 65.

Table 65: Results nf Inuvirm Time~i€ill Assay ei‘ gel and cream farmulatinns against S. aareas MTCC 69%.

Log 1% values {Besil‘lexaeln fermulations) SL7 __GLlS GLlil earn: earn PPtll. . are: Bret V “‘ Time : Wi a l ll name: a (sweat era/ties ca/aaa eL/era— eta/ea (h) EGL/tltlla a a g a a A e 4.? am“ a a til. ________,mmlwm mmwmw___ 0 7.42 7.42 142 142 7.42 7,42 7.42 8.98 9.6a WO 14666 Eaampie 53: In vim? Time-hiii Kinetic Sturiy ai‘ Gei atiaas Laaiieii With Besii’iaxaeinfiCi /' Adapaieae / Cemhinatien af hath against Staphyiacaccus aureus MTCC 6938 i3fi423} Time kili kinetic study (if gel far'muiariens canraining hesifiexacihfiCl or ene or combination 0f both against S. aureus MTCC 6908was performed. The following samples were used far the study, Tahie 66. Exempiar’y farmaiatieas far determining ihe aatihaeteriai activity against S. (garages MTCC 6968 Ier. Farmuiaiian Cerie Betaiis. (lamentsa Na. Cede VLN—F 1 9/8813 Besrfiexacmjfifli arid 1 GL1 3 Adapaiene gei With Carhomer a ADP/GLIGGE4 , Homegaiymer Tyge C ; Besifiexacinl—iCi and VLN—ara/BSF— 2 a GW 3 Adapaieae gel with hydroxyi ADP/GL/QQ3 ethyi eeiiuiase 3 Besifioxacin.HCi (saiuhiiized) VLN-FW/BSF— 1 3 ssar and Adapaiene gel with ADP/GLXOOE h drcx i eth‘i eeiiuiese 4 SL7 VLN-FEl/BSF/GL/GGEA Besrfioxacmgfisaiuhhzed ADGI VLN—F 1 9/ADP/GL/‘001 Gei centaining Adapaiehe a PPQE g/asa/GL/ma-ri Piaeehe gei {@424} Praeedare far In Him Time-Km Assay: Besiflexaein femuiaiiorr was diluted in water to prepare 1 rag/ml stock and further d in BHI broth :0 make final cancenrrarian of 100 gig/min S. aureus MTCC 6908 (900 311) 0.5 McFariand standard equal culture and 100 iii ted Besifloxaein (100 gig/mi) WO 14666 were mixed to obtain final Besifloxacin concentration it) ng/rnl in reaction mixture. Total on mixture (l ml) was incubated at 37°C in an incubator, Post incubation at l, 3 and 6 h, cells were plated after serial dilution in brain heart infusion agar plate. One tube with two read outs were taken for each sample. Cells were allowed to grow for i6—‘24 h at 370C in incubator. {36425} ,iiesnits: All Besifloxacin—Adapalene formulations except placebo had shown similar antibacterial efficacy against S. oureus MTCC 6908 (approximately 2 log reduction at 6 h. Results are shown in Table 67. 'i‘abic 67: Results of Inuvirro Time~i§iii Assa}i of gel iations against S. anreus MTCC 6%8 iii vaiues Besifioxacin ations and their combination with Ada;alone) _ Log 2 nine i Gide sear §E¥““i"”7§bfiim""i" east Time V (Liner Broth (its) gonna: {areas ’ orient nearer, eat ' oblatea a control o 7.53 a 7.53 7.53 1 6°25 l 634 a 3 5-81 IIIIIIIIIIIIIIII * l 824 6 5A8 IIIIIIIIIIIIIIIIIIIIEIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII 0 i 904 094 904 l ooz not cc? l 3 IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII 6 002 005 a 0G5 Example 59: In vitro Time-kill Kinetic Study of iuxacinfitfi‘i Loaded Gel and Cream Formulations against P. (zones CCARli/i 9M9 teens} In vitro 'l‘irne kill kinetic study of gel formulations containing besitloxacin.ilCl against P. genes CCARM §0l0was performed The samples used for the study were same as mentioned in ’i‘abie 64, ititi427} Procedure for inwr’rro ’iiiniewidiii Assay: 0.5 McFarland standard equal P. genes (CCARb/i 9010) culture was ed in sterile water. Prepared culture was centrifuged at 20% rpm for 2G rnin then supernatant was removed and similar amount of brain heart infusion (Bi-ll) broth was added. P. acnes culture in Biii broth was kept for overnight (16 h) incubation in anaerobic box at 37 0C.

Besiiloxacin powder was diluted in water and DMSG to prepare l mgjmi stock and then both were further diluted in Bill broth to make final concentration of 100 gag/ml. 990 ul of P. genes 0.5 McFarland culture (after 16 h. incubation) and 166 at of diluted Besifloxacin (ltll) rig/ml) were mixed to obtain final Besifloxacin concentration 10 ug/ml in reaction mixture. Total reaction mixture (1 ml) was incubated at 37 OC in an anaerobic box. Post incubation at 2, 3 and 24 h, cells were plated after serial dilution in brain heart infusion agar plate, Cells were allowed to grow for 3 clays at 37°C inside anaerobic box. genres} Results: All loxacin formulations except ?lacebo had shown similar antibacterial y t P. acne: CCARM Quill (Approx 2 log it} reduction at 24 hrs): Results are shown in Tables as.

Table 68: Results of In vitro TimeuKill Assay of gel and cream formulations against P. ewes (ICARM illilii 1 Log it} values(Besiiloxaein formulations} e till: In vitre Time—kill Kinetic Study of Besii‘ioxacin.li€l Loaded Gel and (Iream Formulations against P. owes MTtCt: i951 } In vitro Time kill kinetic study of gel and cream formulations containing besifloxacin.llCl or Adapalene or combination of both t P. acnes MTCC lQSlwas performed. The samples used for the study were same as written in Table 54. {66439} Results: All Besiiloxacin formulations except placebo had shown similar antibacterial efficacy against P. genes MTCC l95l (approximately 2 log reduction at 24 it), Results are shown in Table 69.

Tehie 39: Resnits 9f Time-Km Assay 9? get anti cream fermuiatinns against F. 93333 MTCC 1951.

Leg it} veiues (Besit‘iexaein femuintiens) 3““““73-1533:33:33 377.19 329392 .3397.a 3392 THREE "‘5‘ “““3.” ___3—33195.m..___3,_mm”mm 3311311 3.99) 33/9973 33/992333:73 33. 333/9933;Wen/1979-3 3- 339337 9 i 3 3. 79 3.29 3.39 3.19 3.19 3.19 3.79 3.19 2 7.47 745 7.42 7.47 7.43 8.29 3— 3.22 3.23 33.3ng!“ . m594 ““3323”.___-.-m3:99__m3m_§;?lw_3_m_§;§‘§_w_im_3_;§iw 3.35 24 399 .97 3.94 3.94 L 3.99 9.29 9.17 9.39 9.94 9.93 9.99 Examnie fit: Deterntinntien 9f Antiuint’iammatery Petentiai 9f Besit’iexaein.i—i€t, At‘ianaiene in THE—i Ceiis Stimulated by P. ewes (ATCC 6919) 333431} Anti—inflammatery activity of exeein.HCi and adapaiene in THP~ leeiis stimuiated with P. acnes (ATCC 69i 9) was studied.

Preeettnre: {Git/$32} Preparatien ef stimniant fer inflemmatien: P. genes cuiture suspension was prepared in ?BS and the eeii number in the suspensien was adjusted t9 itnateiy SXIQS CiiU/mi by measuring the eeii density using 9 i5 Densirnat. The bacterial suspensien was then heat kiiied at 80°C for 30 min and stered at 980°C nntii further use. 3634333 ELESA t9 study infientntatery resnense in THE-i eeiis: Cells were seeded in a 96—weii fermat (2X 105 THP—i cells per weii) in media eentaining “3% PBS. The cells were ated to induce inflammatory cytokines using 3 2t} MeFariand equivalent heat—killed P. genes. Ceils in controi welis were treated with PBS. One heur after inductien with P. genes. test agents were added tn the induced eeils at appropriate eeneentratiens (Besit‘iexaein at It) and 30 gig/mi; Adapalene at 9.04 and 0.4 gag/ml). The plates were ted at 370C for 24 hours After 24 h, the plates were fuged to pellet the cells and the supernatants were collected. The cell culture supernatants thus ed were analyzed for cytokine levels {lL—d and lL~8) by ELlSA using R&D s kits (A for individual cytoltines following the manufacturer’s instructions.

Results {60434} Besii‘ioxaein inhibits ill-=6 secretion from P. cones—induced ThiP-i cells: in order to study anti-inflammatory effects of hesifloxacin in response to P. l0 acnest THP-l cells were exposed to heat—killed P. acnes followed by treatment with hesitioxaeiniiCl at it) or 3% rig/ml, The culture supernatants were then tested for levels of iL~6 or iL~8 using a colorimetric ELISA. The data ted in Fig 14A clearly shows that oxacintl-lCl causes a significant doseedependent decrease in iL—o levels induced by P. genes. However, it does not show similar l5 effect on iL~8 levels (Fig. 143). The cell viability for each of the tested condition was over 80% compared to untreated cells {data not shown). Dexarnethasonet the known anti—inflammatory agent, used as a positive control, showed nearly 100% reduction in iL~6 levels. ithi435} Combination of adanaiene and hesifiosacin shows additive 2G inhihitory effect on iL—a secretion by P. aches—induced 'i‘iiitmi ceils: The effect of a combination of adapalene and hesifloxacin.llCl on cytokine secretion by THP-i cells was studied, For this purpose ~l. cells were induced with dead P. acnes and treated with hesitioxacin alone (it) rig/mi), adapalene alone (0.94 gig/nil or {3.4 ug/rnl) or hesifloxacinliCl and adapalene combinations at the aforementioned concentrations. The results ed in Fig. 15A Show that at the tested concentrations hoth hesifloxacin.i-iCl and adapalene impart dual anti- inflammatory effects by decreasing the levels of Pr acnesvinduced iLs—é. Further, we observe an ve effect on iL-o reduction when both hesifloxacini-iCl and adapalene are present compared to their individual effects. BesifioxaeinHCl alone 39 at it) gig/ml causes approximately 40% reduction in lL—o levels compared to untreated control. When combined with 0.04 and (14 gig/ml adapalene, the lL—o reductierr goes up to 79% and 80% respectively. These effects were hewever not observed fer P. genes-induced IL—8 levels (Fig. ESE). {88436} Resuits are ted in Tabies ‘78 emf 7i and Figs. 14 and 15.

Tehie 7f}: s ef Effect ef Besifiexaeimfifl en P. genes imiueed eyieiririe (EL — 6. if. - 8) reiease in THE} eeiis as sirewrr in Fig. 14 Expression 0f Cyfekiries Cendifien rife (pg/mi) Lie (g3glmi 'VV V Ceiis + P. genes + BesifloxecirrCi .

J 282.45 13.76 2484.8 169.74 , (if) rig/mi) VVVVV .

Ceiis Jr P. acnes + Besifiexaeini-ECI 4 , ‘ 124.58 19.2 2301.6 133.30 <36 rig/mi) V VV V Ceiis+R acnes+ Bexfimseethane (3.4 ) ' ' 1080 8’ 1929' Ceiis + P. acnes + Vehieie (0.1%) @8437} inferenee: There was significant reduction in P. acnes~indueed IEL~6 i0 ievels pest drug ent. i-iewever there was me reduction in iL-8 ieveis. ”faiiie 7i: Resuiis ef Effect ef Besifiexaeinfifi amiier Adegaierre err P. ewes indeeerfi eytekirre {EL - 6, EL - 8) reiease in ’i‘ffi’hi eeiis es Shawn in Fig. 15 Expressieri ef Cyrekiriesm ,V Sr: Ne. Cemiitierr iL~ 6(grrrri) EL- 8 (gigimi) Cells alone -3.95 V V-ree 81.54 j W21 3412 230-3? ..... 262VVH6 Ceiis + P? acne-s + 2484.76 Besifloxeein.i-i€i (1i) ) Cinde" vvvvvvvvvvvvvvvvvvv ~~~~~~~~~~~~~~ ~ - 7 (0.94 Mm) . . “41.24 .. V - C5135 + P. acne + Aapdiene (NV 162.76 8.45 2579.87 Her/m1) 197.32; WO 14666 Cells + P. genes Besifloxaciniifil l i36.7o i306 2739.20 74.49 (ll) rig/nil) + Adapalene (9‘04 rig/nil) Cells + P. acnes+ Besifioxacin (l0 gig/ml) + Adapalene (9.4 2478.99 65.87 VVVVVVVVVVVVVVVVVVVVVV VV Cells + R ocnes+ Dexamethasone 8 7 1929 o4 rig/ml) .

VnV V V- 193086 Cells 4- P. acnes 4- Vehicle (amt 8 {@6438} inference: l. Besiiloxacini—iCl exerts its anti-inflammatory action by decreasing P. acnes induced lL—6 levels in THP—l cells. 2. Addition of adapalene along with besifloxacinHCl increases the degree of reduction in lL—o levels and offers enhanced anti—inflammatory effect compared to hesifloxacin alone. {@439} All patents and other ations identified in the specification and examples are expressly incorporated herein by reference for all purposes. These if} publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be ued as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the ation available to the l5 ants and does not constitute any ion as to the correctness of the dates or contents of these documents. d} Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, tutions, and the like can be made without departing 26 from the spirit of the invention and these are ore considered to be within the scope of the invention as defined in the claims which follow. Further, to the extent not already indicated, it will be understood by those of ordinary skill in the art that any one of the various embodiments herein described and illustrated can be further modified to incorporate features shown in any of the other embodiments sed herein. it is noted that variations and atiens are pessibie based en the disetesnre above witheut departing from the sprit and scene ef the inventien, REFERENCES 9 Jeremy et a1 (2003). Journal ef Investigative Dennateiegy;121: 20—27; a Thibeut et at (2014). Journai of investigative Dennateiegy;i34: 307L310; e Tagiietti et at (2008) Skin Therapy Lettera 2008,2136): 6—8; a Regnes et ai. (2004) Antimicrob Agents Chemether:48(10): 3670-6; 6 Miller et alt (2004) Scienceg305 : 1629—31; e Keren et at, (2004) J. Bacterioiflgéz 3112—8130; 9 Sehnlzen et a]. (200i), Nature 413: 814~82i§ s Beitrn et a1. (2003) Antimicrob Agents her; 47(3): it 12-4114; 6 Camban et ai. (2009). Antimierebi Chemother.63 (3): 443-450; 8 Kim et at“ (2002) Journai efimmnneiegy; 169(3): 1535-41; 3 Lin et 05) Jennie} ot‘immunniogy; 174(5): 2467-2470; a Nagy et ei.(2006) Mierebes Infect; 8(8): 395-205; 9 Lee et ai. (2010) Arch l Res; 302(10): 74566; 9 Mouser et at. (2003) I invest Dennatei; 121(5): 1226-8; 9 Zastefi’ et at. (2002) Nature; 415: 389—395; e Epand et a}. (1999) Bieehim Biophys Aeta; 1462: 11-28; 6‘ Kabata et at. (1972) erebn Agents Chemether; 2(1): 23~23; and a De Lueca et ai. (2000) Revi Ibeream. Mieni; i7: 116-120

Claims (16)

1. A formulation comprising: (i) 0.5 to 4 (% w/w) oxacin.HCl (Equivalent to Besifloxacin); (ii) 2 to 7 (% w/w) diethylene glycol monoethyl ether; (iii) 0.1 (% w/w) Edetate disodium dehydrate (EDTA); (iv) 2 to 10 (% w/w) glycerin; (v) 0.5 to 5 (% w/w) hydroxyethyl ose; (vi) 0.3 to 1.2 (% w/w) carbomer; (vii) 0.3 to 0.7 (% w/w) yethanol; (viii) 2 to 7 (% w/w) polyethylene glycol 400; (ix) 0 to 1.0 (% w/w) sodium hyaluronate; (x) sodium hydroxide; and (xi) purified water.

2. The formulation of claim 1, wherein the formulation further comprises adapalene.

3. The ation of claim 1, wherein the formulation further comprises a second anti-bacterial agent.

4. The formulation of claim 1 comprising: (i) 1 to 4 (% w/w) Besifloxacin.HCl (Equivalent to Besifloxacin); (ii) 5 (% w/w) diethylene glycol monoethyl ether; (iii) 0.1 (% w/w) Edetate disodium dehydrate (EDTA); (iv) 5 (% w/w) glycerin; (v) 0.5 to 1.5 (% w/w) hydroxyethyl cellulose; (vi) 0.3 to 1.2 (% w/w) carbomer; (vii) 0.7 (% w/w) phenoxyethanol; (viii) 5 (% w/w) polyethylene glycol 400; (ix) 0 to 1 (% w/w) sodium hyaluronate; (x) sodium hydroxide; and (xi) ed water.

5. The formulation of claim 1 comprising: (i) 1 to 2 (% w/w) Besifloxacin.HCl (Equivalent to Besifloxacin); (ii) 5 (% w/w) diethylene glycol monoethyl ether; (iii) 0.1 (% w/w) Edetate disodium dehydrate (EDTA); (iv) 5 (% w/w) glycerin; (v) 0.5 to 1.5 (% w/w) hydroxyethyl cellulose; (vi) 0.3 to 1.2 (% w/w) carbomer; (vii) 0.7 (% w/w) phenoxyethanol; (viii) 5 (% w/w) hylene glycol 400; (ix) 0 to 0.2 (% w/w) sodium hyaluronate; (x) sodium hydroxide; and (xi) purified water.

6. The formulation of claim 1, which is a formulation selected from formulations GL27, GL28, GL29, GL30 and GL31 as shown in Table 1: Table 1: Chemical Name Composition (% w/w) GL27 GL28 GL29 GL30 GL31 Besifloxacin.HCL (Equivalent to oxacin 1 1 2 4 1 Diethylene glycol monoethyl ester 5 5 5 5 5 Edetate disodium dehydrate (EDTA) 0.1 0.1 0.1 0.1 0.1 Glycerin 5 5 5 5 5 Hydroxyethyl cellulose 1.5 1.5 0.5 1.0 0.8 Carbomer 0.7 0.3 1.2 0.4 0.8 yethanol 0.7 0.7 0.7 0.7 0.7 hylene glycol 400 5 5 5 5 5 Sodium hyaluronate 1.0 0.2 0 0.2 0.4 Sodium hydroxide solution q.s. q.s. q.s. q.s. q.s. ed water q.s. q.s. q.s. q.s. q.s.

7. The formulation of claim 6, wherein the formulation is selected from GL27, GL28 and GL31.

8. A method of preparing a medicament comprising an effective amount of a formulation of any one of claims 1-7 adapted for administering against bacterial infection.

9. The method of claim 8, wherein the bacterial infection is by an antibiotic-resistant bacterial strain.

10. The method of claim 8, wherein bacterial infection is by P. acnes.

11. The method of claim 10, wherein P. acnes is a drug resistant strain.

12. The method of claim 10, wherein the drug resistant acne is not responding to a therapeutic amount of clindamycin, minocycline, doxycycline, ycline or erythromycin.

13. Use of a ation of any one of claims 1-7 for the preparation of a medicament for the treatment of acne.

14. Use of claim 13, wherein the acne is drug-resistant acne.

15. Use of any one of claims 13-14, where P. acnes is present and is not ding to a therapeutic dose of clindamycin, minocycline, doxycycline, tetracycline or erythromycin.

16. A formulation selected from formulations GL1, GL2, GL4, CM1, S1, S5, S6, FW1, FW5, FW6, SB1, SB5, SB6, BW1, BW5, BW6, L1, L5, L6, SC1-SC5, GL5-GL7, CM2-CM4, SL1- SL3, CM5, SL4-SL9, GL10-GL13, L17, GA5 and GL19-GL26 as shown in Tables 2- Table 2: Composition (%) GL1 GL2 GL4 CM1 Sr. No. Ingredient 1 Water q.s q.s q.s q.s 2 Carbopol 940 1 1 3 ol 980 NF 0 0 0.8 0.6 Hydroxy Propyl 4 Cellulose-H 0 0 1 0 5 Allantoin 0.2 0.2 0 0 Besifloxacin HCl 6 (equivalent to 1 (D1) 1 (D1) 1 1 besifloxacin) 7 Adapalene 0.1 0 0 0 8 Triethanolamine 1 1 0 0 9 Sodium hydroxide 0 0 0.15 0.3 10 Glycerol 5 5 0 5 11 Propylene Glycol 5 5 0 0 12 PEG 400 5 5 0 0 13 Poloxamer 407 0.2 0.2 0 0 Sod. Lauryl Sulphate 14 Sulphate 0 0 1.6 0 15 Tween 80 0 0 8 0 16 Tween 20 0 0 4 0 diethylene glycol 17 monoethyl ether 0 0 15 0 18 Cetyl Alcohol 0 0 0 2 19 Light Liquid Parafin 0 0 0 5 20 Cyclopentasiloxane 0 0 0 5 21 Steareth 2 0 0 0 2 22 Steareth 21 0 0 0 2 23 BHT 0 0 0 0.1 24 Disodium EDTA 0.1 0.1 0.05 0 25 Phenoxyethanol 0.5 0.5 0.6 0.5 Table 3: Composition (%) S. No. Ingredient S1 S5 S6 1 Water q.s q.s q.s 2 Sodium hydroxide (18% aq.) q.s q.s q.s 3 PEG 1450 2 2 2 4 Methyl Gluceth-20 2.5 2.5 2.5 5 Glycerin 1 1 1.5 6 Besifloxacin 1 1 1 7 Adapalene 0.1 0 0 8 Isopropyl alcohol 20 20 0 9 lene glycol monethyl ether 1 1 1 10 Propylene glycol 1.5 1.5 1.5 11 Ethyl alcohol 0 0 20 12 N-methyl 2-pyrrolidone 3 0 0 13 Salicylic acid 0 0 2 14 Sodium hydroxide q.s q.s q.s 15 Phenoxyethanol 1 1 1 16 Fragrance 0.4 0.4 q.s. Table 4: Composition (%) S. No. Ingredient FW1 FW5 FW6 1 Water q.s. q.s. q.s. 2 Carbopol aqua SF-1 6 6 6 Sodium C14-16 Olefin Sulfonate 3 35 35 35 Sulfonate 4 Sodium lauryl ether te 2 2 2 5 Sodium hydroxide (18% aq.) q.s. q.s. q.s. 6 Cocamidopropylbetaine (30%) 10 5 7 7 um EDTA 0.1 0.1 0.1 8 Glycerin 5 5 3 9 Besifloxacin 1 1 1 10 Adapalene 0.1 0 0 11 N-methyl 2-pyrrolidone 3 0 0 12 Salicylic Acid 0 0 1 13 ene glycol 0 0 4 14 PEG-7 glycerylcocoate 1 1 1 15 Citric Acid (50%) q.s. q.s. q.s. Table 5: Composition S. No. Ingredient SB1 SB5 SB6 1 Sodium palmitate 94.2 94.2 94.2 2 Sodium lauryl ether sulphate 2 2 2 3 Polyquaternium-39 1 1 1 4 Methyl Gluceth-20 1 1 1 5 Titanium dioxide 0.5 0.5 0 6 Besifloxacin 1 1 1 7 Adapalene 0.1 0 0 8 Salicylic acid 0 0 0.1 9 Oleyl oleate 0.5 0.5 0.5 10 BHT (Butylated Hydorxy Toluene) 0.01 0.01 0.01 Table 6: Composition (%) S. No. ient BW1 BW5 BW6 1 Water q.s. q.s. q.s. 2 Disodium EDTA 0.1 0.1 0.1 3 Carbopolaqua SF-1 1 1 1 Ammonium lauryl sulphate 4 (30%