Treatment of resistant acne

Abstract

The present disclosure relates generally to novel molecules, compositions and formulations for the treatment of bacterial infections in general, in particular of antibiotic resistant pathogens.

Description

[0001] This application is a divisional application of patent application No. 201580016977.5, filed on January 29, 2015, entitled "Treatment of Resistant Acne".

[0002] Related applications

[0003] This application claims priority to Indian Patent Application No. 269 / DEL / 2014, filed January 29, 2014, and Indian Patent Application No. 3247 / DEL / 2014, filed November 10, 2014, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0004] This disclosure generally relates to novel molecules, compositions, and formulations for the treatment of bacterial infections in the general sense, particularly bacterial infections caused by antibiotic-resistant pathogens. Background Technology

[0005] Acne vulgaris affects more than 85% of all human skin conditions. Acne is the medical term for clogged pores that typically occur on the face, neck, and upper torso. The following are four main factors known to contribute to the formation of acne vulgaris: (1) increased sebum production resulting in oily, greasy skin; (2) increased bacterial activity, usually attributed to an overgrowth of Propionibacterium acnes; (3) blockage of the hair follicle or pilosebaceous duct (hyperkeratosis); and (4) inflammation. Clogged pores lead to blackheads, whiteheads, papules, or deeper bumps (such as cysts or nodules). Severe cases of acne can result in permanent scarring or disfigurement.

[0006] While acne vulgaris is multifactorial, symbiotic skin bacteria (Propionibacterium acnes) play a major role in the formation of acne lesions. It is an infection of the pilosebaceous sebaceous glands, the oil glands in the skin. In most cases, a sudden acne breakout may be associated with a sudden increase in sebum production in the affected individual. During puberty, androgens play a crucial role. They cause the pilosebaceous sebaceous glands to produce excess sebum. This condition is further exacerbated by the irregular shedding of dead skin cells from the lining of the hair follicle. As dead skin cells clump together in the oily environment, they can form plugs that block the pores of the hair follicle. Pores blocked by shed skin are called comedones (whiteheads).

[0007] This creates a highly favorable anaerobic environment for the growth of *Propionibacterium acnes*. Excessive proliferation of *Propionibacterium acnes* leads to the destruction of the hair follicle wall and sends dangerous signals to the host immune system. *Propionibacterium acnes* can trigger innate immune responses in both very early (microcomedogenic) and late (inflammatory) acne lesions by activating Toll-like receptor 2 (TLR2) on innate immune cells. TLR activation ultimately triggers the expression of various cytokines (such as IL-6, IL-8, IL-12, and IL-17) and chemokines that stimulate the recruitment of other host immune cells [Jeremy et al., 2003; Thibout et al., 2014]. Acne lesions, depending on severity, include blackheads, whiteheads, and papules; as well as more severe lesions such as deeper lumps, cysts, and nodules.

[0008] Although a variety of over-the-counter products are commercially available to alleviate acne symptoms (such as topical anti-acne agents including salicylic acid; sulfur; lactic acid; glycolic acid; pyruvate; urea; resorcinol; N-acetylcysteine; retinoic acid; isotretinoin; tretinoin; adapalene; tazoretene; antibacterial agents such as clindamycin, tetracycline, and erythromycin; vitamins such as folic acid and nicotinamide; minerals such as zinc; benzoyl peroxide; octopirox; triclosan; azelaic acid; phenoxyethanol; phenoxypropanol; and flavonoids), these agents often lack the potential to alleviate acne symptoms and may have negative side effects when designed as routine topical formulations. A major challenge limiting the use of topical formulations is the lack of formulations with ideal physicochemical properties and high drug loading that maintain concentrations significantly above the MIC at the application site by promoting appropriate levels of penetration over time, while having minimal systemic exposure. Formulations targeting these unmet needs could represent a significant advance in the treatment of acne.

[0009] Furthermore, as described in [Taglietti et al., 2008], effective topical formulations can be among the most challenging products to develop when it comes to delivering drugs to specific sites. Once a product is applied to the skin, complex interactions occur between the formulation, the active compound, and the skin itself. The active compound penetrates the skin according to Fick's first diffusion law, which describes the rate of solute transport as a function of the concentration of various components, the size of the treatment surface area, and skin permeability. However, skin permeability can be influenced by a variety of factors, such as dryness, moisturizing, or the blocking effect of excipients in the formulation, where a combination of these factors can modulate the release of the product at the treatment site. In the case of acne, the site of action is within the pilosebaceous unit; therefore, an effective anti-acne formulation should facilitate the penetration of the active compound into this extremely lipophilic environment. Thus, an effective topical formulation needs to provide a stable chemical environment in a suitable dispensing container to accommodate a variety of compounds that may have different (in the absence of incompatibility) physicochemical properties [Taglietti et al., 2008]. Once applied, topical formulations must interact with the skin environment, which can affect the release rate of the compound to achieve sufficient skin absorption and produce additional physical effects on the skin, such as dryness, blockage, or moisturization [Tagleitti et al., 2008]. For example, even if an active agent is highly effective and acts via a systemic route, it may behave quite differently when administered topically; that is, it will not exert an effective anti-acne therapeutic effect if the desired concentration is not reached in the pilosebaceous unit (or skin). Similarly, molecules or drugs may behave quite differently if formulated with different compositions, as will be demonstrated in later examples. Similarly, two molecules or active agents may behave quite differently in the same formulation or composition. Therefore, each new molecule that needs to be formulated for topical skin application presents a new and independent challenge because it is impossible to predict which composition and which ratio of active agent to excipient will provide the desired efficacy benefit.

[0010] Furthermore, a new condition is the evolution of *Propionibacterium acnes* strains that are unresponsive to currently approved antibiotics used to treat acne, such as clindamycin, tetracycline, and erythromycin. Earlier teachings attributed antibiotic failure to the selection of “resistant” strains—that is, mutations that alter the antibiotic’s target, rendering it ineffective. However, new evidence suggests that antibiotic failure is more complex than this simple understanding. The hypothesis was that if resistance develops, i.e., the antibiotic’s target is altered, it might be possible to treat the condition by switching to another antibiotic whose target remains intact in the bacteria. However, recent knowledge has suggested that this assumption is incorrect. For example, Regoes et al. (2004) showed that even in the absence of any resistance, subsets of bacteria can exhibit resistance to antibiotics simply by not undergoing lysis. This can occur due to the physiological (metabolic) and morphological changes observed in bacteria exposed to antibiotics. For example, a study published in Science [Miller et al., 2004] showed that transient induction of SOS in response to ampicillin can protect *E. coli* against the bactericidal effect of ampicillin. Regoes et al., 2004, proposed that tolerance mechanisms may cross between some antibiotics; that is, antibiotic A may become ineffective due to the development of resistance, but it is possible that antibiotic B, which has completely different targets / mechanisms of action and has proven active in different or susceptible bacterial strains, may also become ineffective in resistant strains due to shared tolerance mechanisms. Indeed, significant changes in gene expression have been observed leading to alterations in protein synthesis, stress response pathways, and cell division in *E. coli* during exposure to ampicillin and ofloxacin, and many of these changes at this gene expression level are shared between bacteria exposed to ampicillin and ofloxacin, suggesting that bacteria that do not respond to ampicillin may also not respond to ofloxacin, despite the two drugs having different targets. We saw similar observations when screening antibiotic libraries against different strains of Propionibacterium acnes (which may or may not respond to clindamycin (lincosamide)). Figure 1AAs shown, *Propionibacterium acnes* strains that do not respond to clindamycin also exhibit increased survival in the presence of roxithromycin (a macrolide), which targets a different site than clindamycin [Keren et al., 2004]. Specialized surviving cells and multidrug resistance mechanisms in *Escherichia coli* (J. Bacteriol, 186: 8172-8180) suggest that random fluctuations in gene expression lead to the formation of specialized surviving cells. Phenotypic resistance to antibiotics, as reported by Regoes et al., 2004, can effectively prevent clearance. As a result, while there remains a need in the art for compositions, formulations, and methods for treating acne that does not respond to currently used agents (particularly clindamycin, minocycline, erythromycin, or doxycycline), the probability of resistance makes it difficult to predict which drugs might be effective against *Propionibacterium acnes*.

[0011] Furthermore, it is becoming increasingly apparent that subtle changes in the chemical structure of a molecule can significantly alter its molecular activity against a target protein. For example, erythromycin (a macrolide) and clindamycin bind to similar 50S ribosomal units, but crystal structures [Schulzen et al., 2001] show different binding patterns between the agent and the amino acid residues present in the 50S ribosomal subunit. There are known clindamycin-resistant strains of *Propionibacterium acnes*, but they may be unresponsive to or susceptible to erythromycin, and vice versa. Interestingly, telithromycin (a semi-synthetic derivative of erythromycin) is highly effective in bacterial strains that are simultaneously resistant to erythromycin and clindamycin [Beitru et al., 2003]. Similarly, in another instance, the introduction of an 8-chloro group significantly enhances the potency of moxifloxacin, but similar changes in gatifloxacin are ineffective against *Staphylococcus aureus*, *Streptococcus pneumoniae*, and *Escherichia coli*. Moreover, the same class of molecules can have different affinities for the same protein targets in different bacteria. For example, it has been found that in Staphylococcus aureus, both besifloxacin and moxifloxacin bind to DNA gyrase more effectively than ciprofloxacin. Conversely, in Escherichia coli, ciprofloxacin binds to DNA gyrase more effectively than moxifloxacin or besifloxacin. Similarly, besifloxacin has been found to be the most effective molecule against Streptococcus pneumoniae, followed by moxifloxacin and ciprofloxacin [Cambau et al., 2009]. Therefore, it is impossible to predict the activity of a molecule against bacteria or microorganisms based on its structural similarity to another drug that shows activity against the same or different microorganisms, even if they may have similar mechanisms of action. In fact, as shown in Figure 1, we observed that structurally very similar molecules have completely different activities against Propionibacterium acnes, i.e., one is inactive against clindamycin-sensitive and non-resistant Propionibacterium acnes, while the other is very effective. Figure 1A and Figure 1BIn another example, which we will discuss later, we observed that nonlincosamide molecules were highly effective in clindamycin-resistant Propionibacterium acnes strains but inactive in clindamycin-sensitive Propionibacterium acnes. Figure 1A and Figure 1B Therefore, during the systematic screening of Propionibacterium acnes, an unexpected discovery led to the identification of effective drugs that work against sensitive Propionibacterium acnes as well as those that are unresponsive to clindamycin, minocycline, erythromycin, or doxycycline.

[0012] Furthermore, while the problem lies in the development of microbial resistant strains that do not respond to compositions and compounds known in the art, there remains a need for more effective antibiotics that not only act on resistant microorganisms but also reduce the risk of microorganisms developing resistance to the new antibiotic. Therefore, molecules that are effective antibiotics and also “prevent” or reduce the development of resistance could be a major advance in the treatment of microbial diseases.

[0013] The inflammatory features of acne have been associated with host immune responses targeting *Propionibacterium acnes*. In vitro studies have shown that whole cells or fragments of *Propionibacterium acnes* stimulate immune cells, keratinocytes, and sebaceous cells to release cytokines and matrix metalloproteinases [Kim et al., 2002; Liu et al., 2005; Nagy et al., 2006; Lee et al., 2010]. Although *Propionibacterium acnes* is present in the follicular region long before follicle rupture, it only comes into direct contact with immune cells in the dermis after follicle rupture. The innate immune system recognizes *Propionibacterium acnes* via TLR2 [Kim et al., 2002], leading to the secretion of inflammatory cytokines, including IL-6, IL-8, and IL-12. Follicle rupture occurs at a very late stage of the disease process. However, there is also evidence that adaptive immune responses play a significant role even in the inflammation observed in the early stages of acne due to the recruitment of activated T helper 1 (Th1) lymphocytes to early acne lesions [Mouser et al., 2003]. Therefore, potential acne treatments need to address inflammation and should be able to target these inflammatory pathways.

[0014] Therefore, ideal acne treatment requires molecules that target two or more substances. The following molecules can be powerful strategies for treating acne: molecules that are effective against antibiotic-sensitive Propionibacterium acnes strains and strains resistant to or unresponsive to clindamycin, minocycline, erythromycin, and doxycycline, and that also inhibit inflammatory mediators activated by Propionibacterium acnes; or molecules that target two or more of these microorganisms while simultaneously inhibiting inflammatory mediators activated by Propionibacterium acnes, and are formulated to achieve the desired concentration of active agent in the skin or pilosebaceous region after topical application. Summary of the Invention

[0015] DART

[0016] A new series of DART (Dual-Action Rational Therapy) molecules have been designed and synthesized for the treatment of bacterial infections caused by susceptible Gram-positive and Gram-negative bacteria, particularly for the treatment of acne and infections of various skin and skin structures, while also preventing the development of resistance. DART molecules can enhance their activity in microorganisms (such as bacteria) through two distinct mechanisms of action, and offer less opportunity for the development of mutations at two target sites in bacteria. Furthermore, they can also exert their effects at the host level by modulating the immune response, such as altering the levels of inflammatory cytokines.

[0017] DART is designed with two active domains. These domains can be selected from different families, such as β-lactams, β-lactam derivatives, 2-quinolones and 4-quinolones, quinolones with a halogenated atom (especially a fluorine atom) attached to the C-6 or C-7 position of the central ring system, fluoroquinolones with a halogenated atom (especially a chlorine atom) attached to the C-8 position of the central ring system, tetracyclines, oxazolidinones, hydroxypyridinones, hydroxypyridinone derivatives, truncated pleurotin, pyrroles, nitroimidazoles, monooxycarbolic acids, fusidic acid, sulfonamides, sulfonamide derivatives, retinoids, different fatty acids (saturated and unsaturated), propylene glycol and glycerol derivatives of different fatty acids, and strategic combinations of various families. Strategically, the two active domains are arranged in a spatially correct manner for both domains to maintain their antibacterial or antifungal function. In summary, these molecules exhibit faster bactericidal activity as inflammation subsides and are active against resistant pathogens. These molecules also show a lower risk of resistance development.

[0018] In some embodiments, the DART molecule has at least two chemical domains. Each of the chemical domains binds to a distinct or different active site in the target cell. In a preferred embodiment, a third chemical domain may be present. In a further preferred embodiment, the two chemical domains can be linked together through the third domain. In some embodiments, the DART molecule has at least two distinct or different antibacterial mechanisms of action. In some embodiments, the DART molecule has at least two distinct or different anti-acne mechanisms of action. Without limitation, DARP may act on the same target or on different targets, such as bacteria and the host. In some embodiments, DART acts on at least two different targets. In some embodiments, at least one of the targets is different from a target affected by conventional antibiotics.

[0019] In some embodiments, the DART molecule has a β-lactam ring and a quinolone core, or a quinolone core and a nitro heterocycle, or a β-lactam ring and a nitro heterocycle.

[0020] In some embodiments, DART has at least two distinct antibacterial mechanisms of action, such as inhibition of DNA gyrase or topoisomerase IV and transpeptidase-mediated peptidoglycan crosslinking, inhibition of isoprenyl pyrophosphate and transpeptidase-mediated peptidoglycan crosslinking, inhibition of isoprenyl pyrophosphate and topoisomerase IV DNA gyrase, inhibition of folic acid synthesis and topoisomerase IV DNA gyrase, inhibition of folic acid synthesis and transpeptidase-mediated peptidoglycan crosslinking, inhibition of DNA gyrase or topoisomerase IV and the 30S ribosomal subunit in bacteria, and inhibition of DNA gyrase. The enzyme or topoisomerase IV inhibits the 50S subunit in bacteria, or inhibits transpeptidase-mediated peptidoglycan cross-linking and the 30S or 50S ribosomal subunit in bacteria; or inhibits folic acid synthesis and the 30S or 50S subunit in bacteria; or inhibits isoprenyl pyrophosphate and the 30S or 50S subunit in bacteria; or induces DNA modification, such as induced DNA nicks, while inhibiting the induction of DNA negative supercoiling; or alters cell membrane fluidity while exerting activity on DNA; or alters the level of metal ions in the cell while inducing DNA changes. In some embodiments, the first mechanism of action is antibacterial, and the second mechanism of action is anti-inflammatory or immunomodulatory.

[0021] In some embodiments, the DART molecule has at least two distinct mechanisms of action for treating acne and regulates at least two different targets. In some embodiments, the first mechanism is antibacterial action, and the second mechanism is inhibition of keratinocyte proliferation and differentiation. In some embodiments, the DART molecule has two distinct mechanisms of action for treating acne, wherein the first mechanism is antibacterial action and the second mechanism is anti-inflammatory. In some embodiments, the DART molecule is effective against forms of Propionibacterium acnes that are poorly responsive to anti-acne products containing clindamycin, doxycycline, erythromycin, or minocycline. In some embodiments, they are effective against one or more clindamycin-, minocycline-, erythromycin-, and / or doxycycline-resistant strains of Propionibacterium acnes. In some embodiments, it prevents the development of resistance in Propionibacterium acnes.

[0022] In some embodiments, the DART molecule possesses at least two distinct antibacterial mechanisms of action and regulates at least two different targets against the pathogen. A non-limiting example of such a pathogen is Bartonella henselae (…). Bartonella henselae Borrelia burgdorferi () Borrelia burgdorferi Campylobacter jejuni ( Campylobacter jejuni Campylobacter fetus ( Campylobacter fetus ), Chlamydia trachomatis ( Chlamydia trachoma Chlamydia pneumoniae ( Chlamydia pneumoniae ), Chlamydia psittaci ( Chylamydia parrots ), Simkania negevensis Escherichia coli ( Escherichia coli (e.g., O157:H7 and K88), Ericksonites ( Ehrlichia chafeensis Clostridium botulinum ( Clostridium botulinum Clostridium perfringens ( ) Clostridium perfringens Clostridium tetani ( Clostridium tetani ), Enterococcus faecalis ( Enterococcus faecalis Haemophilus influenzae ( ) Haemophilus influenzae ), Haemophilus ducreyi ( Haemophilius ducreyi ), Coccidioides immitis ( Coccidioides immitis Bordetella pertussis ( ) Bordetella whooping cough ), Rickettsia burnetii ( Coxiella burnetii ), Ureaplasma urealyticum ( Ureaplasma urealyticum ), Mycoplasma genitalium ( Mycoplasma genitalium ), Trichomonas vaginalis Helicobacter pylori ( Helicobacter pylori ), Helicobacter hepatis ( Helicobacter hepaticus Legionella pneumophila ( Legionella pneumophila ), Mycobacterium tuberculosis ( Mycobacterium tuberculosis ), Mycobacterium bovis ( Mycobacterium bovis ), Mycobacterium africanum ( Mycobacterium africanum Mycobacterium leprae ( Mycobacterium leprae ), Mycobacterium Asiatum ( Mycobacterium asiaticum ), Mycobacterium avium ( Mycobacterium avium ), Mycobacterium cryptans ( Mycobacterium cryptatum ), Mycobacterium tectorum ( Mycobacterium chelonae Occasionally occurring mycobacteria ( Mycobacterium fortuitum ), Mycobacterium Genevai ( Mycobacterium genavense Haemophilus ( ) Mycobacterium haemophilus Intracellular mycobacteria ( Mycobacterium intracellulare ), Mycobacterium Kansas ( Mycobacterium kansasii ), Mycobacterium marmosetum ( Mycobacterium malmoensis ), Mycobacterium marinum ( Marine Mycobacterium ), Mycobacterium scrofula ( Mycobacterium scrofulaceum Mycobacterium simianum ( Mycobacterium simianum ), Mycobacterium stearothermia ( Mycobacterium szulgai ), Mycobacterium ulcerans ( Mycobacterium ulcerans ), Mycobacterium bufossa ( Mycobacterium xenopis Corynebacterium diphtheriae ( Corynebacterium diphtheriae ), Rhodococcus equi ( Rhodococcus equi ), Rickettsia ehrlichii ( Rickettsia aeschlimannii ), African rickettsia ( Rickettsia africana ), Connorricklet ( Rickettsia conorii Cryptolytica hemolyticus ( Arcanobacterium haemolyticum Bacillus anthracis ( Bacillus anthracis ), Bacillus cereus ( Bacillus cereus Listeria monocytogenes ( ) Listeria monocytogenes Yersinia pestis (Yersinia pestis) Yersinia pestis Yersinia enterocolitica (Yersinia enterocolitica) Yersinia enterocolitica ), Shigella dysenteriae ( Shigella dysenteriae ), Neisseria meningitidis ( Neisseria meningitidis ), Neisseria gonorrhoeae ( Neisseria gonorrhoeae Streptococcus bovis () Streptococcus bovis ), hemolytic streptococci ( Streptococcus hemolyticus Streptococcus mutans () Streptococcus mutans Streptococcus pyogenes ( ) Streptococcus pyogenes Streptococcus pneumoniae () Streptococcus pneumoniae Staphylococcus aureus ( Staphylococcus aureus Staphylococcus epidermidis ( Staphylococcus epidermidis Staphylococcus pneumoniae () Staphylococcus pneumoniae ), saprophytic Staphylococcus ( Staphylococcus saprophyticus ), Vibrio cholerae ( Cholera vibrio ), Vibrio parahaemolyticus ( Vibrio parahaemolyticus ), Salmonella typhi Salmonella typhi Salmonella paratyphi () Salmonella paratyphoid Salmonella enteritidis ( ) Salmonella enteritidis Treponema pallidum (Syphilis) Treponema pallidum Candida ( White Cryptococcus ( Cryptococcus Cryptosporidium ( Cryptosporidium Giardia lamblia ( Giardia lamblia Microsporidia ( Microsporidia ), Plasmodium vivax ( Plasmodium vivax Pneumocystis carinii ( Pneumocystis Carinian ), Toxoplasma gondii ( Toxoplasma gondii ), Trichophyton mentagrophytes ( Trichophyton mentagrophytes ), Intestinal microsporidia ( Enterocytozoon bieneusi Cyclospora ( Cyclospora cayetanensis ), Helen's encephalitis microsporidia ( Encephalitozoon hellem Rabbit intracellular protozoa ( Encephalitozoon of rabbits ), as well as other bacteria, archaea, protozoa and fungi.

[0023] In some embodiments, the first and second domains independently possess antibacterial activity against the genus Staphylococcus. Examples of the genus Staphylococcus include, but are not limited to, Staphylococcus aureus (e.g., Staphylococcus aureus).S. golden Staphylococcus aureus ( S. simiae Staphylococcus aureus (e.g., Staphylococcus aureus) S. auricularis Staphylococcus aureus (e.g., Staphylococcus aureus) S. carnosus Staphylococcus conti Monti () S. condiments Staphylococcus masei ( S. massiliensis ), fish fermentation Staphylococcus ( S. piscifermentans ), mimicking Staphylococcus ( S. simulans Staphylococcus epidermidis (e.g., Staphylococcus capitella) St. of the head ), Staphylococcus aureus ( S. goats Staphylococcus epidermidis ( S. epidermidis ), Staphylococcus aureus ( S. saccharolyticus ), hemolytic staphylococci (e.g., S. devriesei hemolytic staphylococci ( S. haemolyticus Staphylococcus aureus ( S. of man ), porcine intermediate staphylococci (e.g., Staphylococcus chromogenicus ( S. chromogenes Staphylococcus feli ( ), Staphylococcus feli S. cat Staphylococcus aureus (), Staphylococcus aureus S. delphini ), Staphylococcus aureus ( S. hyicus Staphylococcus intermedia ( S. intermediate ), Staphylococcus aureus ( S. otter Staphylococcus vulgaris ( ), Staphylococcus aureus S. microti ), Staphylococcus aureus ( S. flies ), Pseudomonas intermedia ( S. pseudintermedius ), S. beak Staphylococcus schreiberensis (Schleifera) S. grinders Staphylococcus ludensii (e.g., Staphylococcus ludensii) from Lugdunum ), saprophytic staphylococci (e.g., Staphylococcus alrighti) S. arlettae ), Staphylococcus kohlii ( S. cohnii ), Staphylococcus equi ( S. of horses Staphylococcus aureus () S. hens Staphylococcus krill ( S. kloosii ), S. leei Staphylococcus nebitis ( S. nepalensis ), saprophytic Staphylococcus ( S. saprophyticus Staphylococcus aureus ( S. amber Staphylococcus xylose ( S. xylosus ), and squirrel staphylococci (e.g., S. fleurettii Staphylococcus aureus (Slow-acting Staphylococcus) S. lentus ), Squirrel Staphylococcus ( S. squirrels ), S. stepanovicii Staphylococcus aureus ( S. calf ), mimicking staphylococcal flora (e.g., mimicking Staphylococcus aureus ( S. simulans )) and Staphylococcus wartii (e.g., Staphylococcus pastoris ( St. pastor Staphylococcus warwick ( S. warneri )).

[0024] Without limitation, DART can be in the form of particles, powders, suspensions, dispersions, emulsions, liposomes, microparticles, microspheres, solutions, vesicles, aggregates, creams, gels, etc.

[0025] This disclosure also provides formulations containing DART as an active pharmaceutical ingredient (API).

[0026] antibiotic

[0027] This disclosure also provides formulations comprising non-DART antibiotics. In some embodiments, the antibiotic formulation is an 8-chlorofluoroquinolone. Exemplary 8-chlorofluoroquinolones include, but are not limited to, besifloxacin, clinfloxacin, and sitafloxacin. In some embodiments, the formulation comprises besifloxacin as an API.

[0028] In various embodiments, the API may be micronized, suspended, or solubilized. In some embodiments, the API may be in the form of particles, powders, suspensions, dispersions, emulsions, liposomes, microparticles, microspheres, solutions, vesicles, and aggregates. In some embodiments, the API may be in the form of a drug carrier.

[0029] In some implementations, the API may be coated. In other implementations, the API may be uncoated.

[0030] Non-limitingly, the formulation may be in the form of a product selected from the group consisting of: lotions, creams, gels, latexes, oils, serums, powders, sprays, ointments, solutions, suspensions, dispersants, pastes, foams, release agents, films, masks, patches, sticks, roll-on products, cleansing liquids, cleansing sticks, pastes, foams, powders, shaving creams, and fabric-impregnated products. In some embodiments, the formulation is in the form of a product selected from the group consisting of: gels, creams, sprays, facial cleansers, soap bars, shower gels, lotions, drug-loaded suspensions, drug-loaded suspensions, and any combination thereof.

[0031] In some implementations, the API or formulation can be used to treat acne that is not responsive to antibiotics. Specifically, it is more effective against forms of Propionibacterium acnes that are less responsive to anti-acne products containing clindamycin, doxycycline, erythromycin, or minocycline.

[0032] In some implementations, the API or formulation can be used to treat acne by exerting an anti-inflammatory effect.

[0033] In some implementations, the API or formulation can be used to treat acne by killing one or more strains of Propionibacterium acnes that are sensitive to clindamycin, minocycline, erythromycin and / or doxycycline, and by exerting better efficacy by inhibiting the Propionibacterium acnes-mediated inflammatory pathway (i.e., a dual mechanism of action).

[0034] combination

[0035] This disclosure also provides formulations comprising combinations of two or more antibiotic agents. For example, 8-chlorofluoroquinolones combined with another anti-acne agent. In some embodiments, the formulation comprises two or more different 8-chlorofluoroquinolones. In some embodiments, the formulation comprises besifloxacin and a retinoid, such as adapalene.

[0036] In some embodiments, the formulation comprises an anti-acne agent and an anti-inflammatory agent. For example, the formulation may comprise 8-chlorofluoroquinolone and an anti-inflammatory agent.

[0037] In some embodiments, the two or more antibiotic agents may be DART molecules, or two or more different DART molecules. In some embodiments, one of the two or more antibiotic agents is a DART molecule, and the other is not a DART molecule.

[0038] As described herein, this disclosure provides formulations containing DART and / or non-DART antibiotic agents as APIs. Thus, exemplary APIs for use in formulations include DART, antibacterial agents, antifungal agents, and antiacne agents. In some embodiments, the API may be in the form of a drug carrier; that is, the API may be nanotized, coated, or formulated into vesicles, liposomes, emulsions, etc., for use in formulations. Without limitation, the formulation or composition may be formulated for administration via any suitable route of administration known in the art, including but not limited to local (including oral and sublingual) and oral or parenteral routes (including intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, and nasal administration).

[0039] The DART and formulations disclosed herein can be used to treat bacterial infections attributed to Gram-positive or Gram-negative bacteria. Furthermore, the DART is effective against resistant forms of the pathogen. In addition, the DART effectively prevents the development of resistant forms of the pathogen. Therefore, the DART and formulations disclosed herein can be used to treat antibiotic-resistant or antibiotic-resistant bacterial infections. Exemplary bacterial infections include, but are not limited to, Bartonella henselae (…). Bartonella henselae Borrelia burgdorferi () Borrelia burgdorferi Campylobacter jejuni ( Campylobacter jejuni Campylobacter fetus ( Campylobacter fetus ), Chlamydia trachomatis ( Chlamydia trachomatis Chlamydia pneumoniae ( Chlamydia pneumonia ), Chlamydia psittaci ( Chylamydia psittaci ), Simkania negevensis Escherichia coli ( Escherichia coli (e.g., O157:H7 and K88), Ericksonites ( Ehrlichia chafeensis Clostridium botulinum ( Clostridium botulinum Clostridium perfringens ( ) Clostridium perfringens Clostridium tetani ( Clostridium tetani ), Enterococcus faecalis ( Enterococcus faecalis Haemophilus influenzae ( ) Haemophilus influenzae ), Haemophilus ducreyi ( Haemophilius ducreyi ), Coccidioides immitis ( Coccidioides immitis Bordetella pertussis ( ) Bordetella pertussis ), Rickettsia burnetii ( Coxiella burnetii ), Ureaplasma urealyticum ( Ureaplasma urealyticum ), Mycoplasma genitalium ( Mycoplasma genitals ), Trichomonas vaginalis Helicobacter pylori ( Helicobacter pylori ), Helicobacter hepatis ( Helicobacter hepaticus Legionella pneumophila ( Legionella pneumophila ), Mycobacterium tuberculosis ( Mycobacterium tuberculosis ), Mycobacterium bovis ( Mycobacterium bovis ), Mycobacterium africanum ( Mycobacterium africanum Mycobacterium leprae ( Mycobacterium leprae ), Mycobacterium Asiatum ( Mycobacterium asiaticum ), Mycobacterium avium ( Mycobacterium avium ), Mycobacterium cryptans ( Mycobacterium cryptatum ), Mycobacterium tectorum ( Mycobacterium chelonae Occasionally occurring mycobacteria ( Mycobacterium fortuitum ), Mycobacterium Genevai ( Mycobacterium genavense Haemophilus ( ) Mycobacterium haemophilus Intracellular mycobacteria ( Mycobacterium intracellulare ), Mycobacterium Kansas ( Mycobacterium kansasii ), Mycobacterium marmosetum ( Mycobacterium malmoensis ), Mycobacterium marinum ( Marine Mycobacterium ), Mycobacterium scrofula ( Mycobacterium scrofulaceum Mycobacterium simianum ( Mycobacterium simianum ), Mycobacterium stearothermia ( Mycobacterium szulgai ), Mycobacterium ulcerans ( Mycobacterium ulcerans ), Mycobacterium bufossa ( Mycobacterium xenopis Corynebacterium diphtheriae ( Corynebacterium diphtheriae ), Rhodococcus equi ( Rhodococcus equi ), Rickettsia ehrlichii ( Rickettsia aeschlimannii ), African rickettsia ( Rickettsia africana ), Connorricklet ( Rickettsia conorii Cryptolytica hemolyticus ( Arcanobacterium haemolyticum Bacillus anthracis ( Bacillus anthracis ), Bacillus cereus ( Bacillus cereus Listeria monocytogenes ( ) Listeria monocytogenes Yersinia pestis (Yersinia pestis) Yersinia pestis Yersinia enterocolitica (Yersinia enterocolitica) Yersinia enterocolitica ), Shigella dysenteriae ( Shigella dysenteriae ), Neisseria meningitidis ( Neisseria meningitidis ), Neisseria gonorrhoeae ( Neisseria gonorrhoeae Streptococcus bovis () Streptococcus bovis ), hemolytic streptococci ( Streptococcus hemolyticus Streptococcus mutans () Streptococcus mutans Streptococcus pyogenes ( ) Streptococcus pyogenes Streptococcus pneumoniae () Streptococcus pneumoniae Staphylococcus aureus ( Staphylococcus aureus Staphylococcus epidermidis ( Staphylococcus epidermidis Staphylococcus pneumoniae () Staphylococcus pneumoniae ), saprophytic Staphylococcus ( Staphylococcus saprophyticus ), Vibrio cholerae ( Cholera vibrio ), Vibrio parahaemolyticus ( Vibrio parahaemolyticus ), Salmonella typhi Salmonella typhi Salmonella paratyphi () Salmonella paratyphoid Salmonella enteritidis ( ) Salmonella enteritidis Treponema pallidum (Syphilis) Treponema pallidum Candida ( White Cryptococcus ( Cryptococcus Cryptosporidium ( Cryptosporidium Giardia lamblia ( Giardia lamblia Microsporidia ( Microsporidia ), Plasmodium vivax ( Plasmodium vivax Pneumocystis carinii ( Pneumocystis carinii ), Toxoplasma gondii ( Toxoplasma gondii ), Trichophyton mentagrophytes ( Trichophyton mentagrophytes ), Intestinal microsporidia ( Enterocytozoon bieneusi Cyclospora ( Cyclospora cayetanensis ), Helen's encephalitis microsporidia ( Encephalitozoon hellem Rabbit intracellular protozoa ( Encephalitozoon cuniculi Other bacteria, archaea, protozoa, and fungi are also present. In some embodiments, infection occurs with Staphylococcus species.

[0040] In some embodiments, the DART and formulations disclosed herein can be used to treat acne. In some embodiments, the DART and formulations disclosed herein are effective against forms of Propionibacterium acnes that are poorly responsive to anti-acne products containing clindamycin, or doxycycline, or erythromycin, or minocycline. In some embodiments, the DART and formulations disclosed herein are effective against strains of Propionibacterium acnes that are resistant or tolerant to one or more of clindamycin, minocycline, erythromycin, and / or doxycycline. For treatment of infection, the DART or formulations disclosed herein may be administered to the subject as a single dose or multiple doses once or daily. Attached Figure Description

[0041] Figure 1A and Figure 1B The dose-response curves of different antibiotics against Propionibacterium acnes strains MTCC 1951 and CCARM 9010 are shown. Although MTCC 1951 was killed by clindamycin, the drug had no effect on CCARM 9010. The effects of different antibiotics on different strains belonging to different families of clindamycin are different and unpredictable.

[0042] Figure 1C and Figure 1D This demonstrates the effects of DART compounds 90, 91, 94, 113, 115, and 116 on clindamycin-sensitive (MTCC 1951) ( Figure 1C () and unresponsive to clindamycin (CCARM 9010) () Figure 1D Linear graphs of concentration-efficacy curves in *Propionibacterium acnes* strains. The molecules exhibit different and unpredictable activities against *Propionibacterium acnes* strains MTCC 1951 and CCARM9010. Compound 91 showed a highly effective bactericidal spectrum against both bacterial strains. Compound 90 showed activity against *Propionibacterium acnes* strains that are not responsive to clindamycin, but was inactive against *Propionibacterium acnes* strains responsive to clindamycin.

[0043] Figure 2A and Figure 2B The concentration-dependent inhibition of DNA helicase activity (supercoiling) by compound 91 is shown. Figure 2A: Agarose gel electrophoresis shows the effect of compound 91 on the supercoiling of E. coli plasmid DNA caused by DNA helicase. Figure 2B: Percentage of DNA supercoiling caused by DNA gyrase in the presence of compound 91 at increased concentrations.

[0044] Figure 3A The bar chart shows the percentage of DNA supercoiling caused by DNA gyrase in the presence of relaxed *E. coli* plasmid DNA and compounds 90, 91, 94, 113, 115, and 116. Of all the comparisons, compounds 91 and 116 appear to have the best gyrase inhibitory activity. While this observation is primarily correlated with MIC data against *Propionibacterium acnes*, some species-specific advantages were observed for compound 91.

[0045] Figure 3B The bar chart shows the percentage of DNA supercoiling caused by DNA gyrase in the presence of relaxed E. coli plasmid DNA, as well as naflufloxacin and compound 91. Compound 91 showed better potency than naflufloxacin.

[0046] Figure 4A and Figure 4B The bar chart shows the effects of compound 91 on Propionibacterium acnes-induced cytokines IL-6 (Figure 4A) and IL-8 (Figure 4A). Figure 4B The effect of compound 91 on release in THP-1 cells. Compound 91 exerts anti-inflammatory activity against the production of cytokines induced by Propionibacterium acnes. Statistical analysis was performed using the Student t-test. p=0.05; p=0.005).

[0047] Figure 5A and Figure 5B The bar chart shows the effects of compound 91 on Propionibacterium acnes-induced cytokines IL-1α (Figure 5A) and IL-2β (Figure 5B). Figure 5B The effect of release in THP-1 cells.

[0048] Figure 6 The bar chart illustrates the minimum inhibition values ​​of some exemplary topical gel formulations against Propionibacterium acnes.

[0049] Figure 7 Line graphs showing dose-response curves of the zone of inhibition (ZOI) of Propionibacterium acnes for some exemplary topical gel formulations are presented.

[0050] Figure 8 Linear graphs are shown illustrating the time-dependent bactericidal kinetics of some exemplary gel formulations against Propionibacterium acnes.

[0051] Figure 9The graphs illustrate the efficacy of besifloxacin topical formulations against Propionibacterium acnes in an in vivo skin infection model. Besifloxacin gel formulations demonstrated the ability to clear nearly 1.5 log CFU (~95%) of clindamycin-resistant Propionibacterium acnes inoculum within the first 24 hours.

[0052] Figure 10 Linear graphs showing the time-kickdown kinetics of some exemplary besifloxacin formulations against Propionibacterium acnes MTCC 1951 are presented, illustrating how the composition of the formulation can alter the efficacy of the antibiotic.

[0053] Figure 11 To illustrate some exemplary besifloxacin formulations and besifloxacin API against Staphylococcus aureus (Staphylococcus aureus) S. aureus Linear graph of time-dependent sterilization kinetics.

[0054] Figure 12 Linear graphs showing the time-dependent bactericidal kinetics of besifloxacin against Propionibacterium acnes (CCARM 9010) are presented.

[0055] Figure 13 The graphs illustrate how different excipient compositions in topical formulations of the same antibiotic can result in different profiles in the skin and systemic circulation of SD rats. For comparative purposes, completely suspended 1% besifloxacin gel (VLN-F19 / BSF / GL / 068), completely soluble 1% besifloxacin gel (VLN-F21 / BSF / GL / 001A), and completely suspended 1% besifloxacin gel (VLN-F20 / BSF / CR / 004) were used. For efficacy, the formulation should not only be physicochemically compatible with the antibiotic but also maintain antibiotic concentrations above the MIC level.

[0056] Figure 14A and Figure 14B The bar chart shows the effect of becifloxacin on Propionibacterium acnes induced by the cytokine IL-6 (Figure 14A) instead of IL-8 (Figure 14A). Figure 14B The concentration-dependent inhibitory effect of [the substance] released in THP-1 cells was analyzed using the Student t-test. p=0.005; p=0.0005).

[0057] Figure 15A and Figure 15B The bar chart shows that the combination of besifloxacin and adapalene increases the potency against the release of the cytokine IL-6 (Fig. 15A) induced by Propionibacterium acnes in THP-1 cells, but not against IL-8 ( Figure 15BRelease in THP-1 cells was ineffective. Statistical analysis was performed using the Student t-test. p=0.005; p=0.001). Detailed Implementation

[0058] Acne vulgaris affects more than 85% of all human skin conditions. Acne is the medical term for clogged pores that typically occur on the face, neck, and upper torso. The following are four main factors known to contribute to the formation of acne vulgaris: (1) increased sebum production resulting in oily, greasy skin; (2) increased bacterial activity, usually attributed to an overgrowth of Propionibacterium acnes; (3) blockage of the hair follicle or pilosebaceous duct (hyperkeratosis); and (4) inflammation. Clogged pores lead to blackheads, whiteheads, papules, or deeper bumps (such as cysts or nodules). Severe cases of acne can result in permanent scarring or disfigurement.

[0059] As shown at http: / / thescienceofacne.com / antibiotic-susceptibility-of-propionibacterium-acnes / , research results over the past four decades clearly demonstrate that *Propionibacterium acnes* bacteria have increasingly developed resistance to certain classes of antibiotics over time. Of particular importance, a significant percentage of bacteria isolated from acne patients have been observed to be resistant to the most commonly used antibiotics in acne treatment (clindamycin, erythromycin, tetracycline, doxycycline, and minocycline). Furthermore, these resistant or antibiotic-resistant strains can cause acne relapses and other disease states. There is a need for antibiotics that can kill *Propionibacterium acnes* while minimizing the possibility of the development of resistant strains or mutants capable of surviving antibiotic exposure, as well as antibiotics that can target strains that do not respond to current drugs. Moreover, if these novel molecules can target additional steps in acne formation, such as inflammation, their clinical efficacy in acne could exceed that of existing therapies.

[0060] The skin is a major organ of the human body and, in addition to acting as a barrier, performs many essential functions, such as maintaining homeostasis. Besides acne, there are many other skin diseases caused by bacterial colonization of the skin. For mild to moderate skin infections (e.g., acute bacterial skin and skin structure infection (ABSSSI)), the most common bacteria is Staphylococcus aureus (…). Staphylococcus ) and Streptococcus ( StreptococcusThese bacteria can infect the skin of both children and adults; they primarily develop during hospitalization or stay in a nursing home, but can also develop while gardening or swimming. Some people are at particular risk of developing a skin infection, such as those with diabetes, HIV or AIDS or other immune disorders, or hepatitis, as well as those undergoing chemotherapy or other medications that suppress the immune system.

[0061] Common bacterial skin infections include cellulitis, erysipelas, impetigo, folliculitis, and boils and carbuncles. Cellulitis is a painful, erythematous infection of the dermis and subcutaneous tissue, characterized by fever, edema, and bordered progression, and is usually caused by Streptococcus or Staphylococcus. Erysipelas is a superficial form of well-defined cellulitis and is almost exclusively caused by Streptococcus. Impetigo is also caused by Streptococcus or Staphylococcus and can lead to lifting of the stratum corneum, producing the common bullous effect. Folliculitis is an inflammation of the hair follicle, and it is most commonly caused by Staphylococcus. If the infection of the hair follicle goes deeper and involves more follicles, it progresses to the boil / carbuncle stage, usually requiring incision and drainage. Due to toxins produced by bacteria, two different types of skin diseases occur, including staphylococcal squamous skin syndrome (SSSS), which typically affects children under 5 years old and adults with kidney failure, and toxic shock syndrome. Staphylococcus aureus has a greater chance of colonizing in patients with eczema and atopic dermatitis (an inflammatory, recurrent, non-contagious pruritus of the skin). Therefore, Staphylococcus aureus infection plays a significant role in atopic dermatitis (AD) or atopic eczema (AE). Unfortunately, some strains of Staphylococcus aureus have developed resistance to methicillin and other similar antibiotics known as MRSA. It has recently been found that more than half of all cases of bacterial skin infections are caused by MRSA species. Infections associated with MRSA cannot be cured with traditional penicillin-related drugs. Instead, MRSA must be treated with alternative antibiotics.

[0062] However, as stated at http: / / thescienceofacne.com / 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 certain types of bacteria, while remaining largely ineffective against others. Furthermore, antibiotic sensitivity and resistance are dynamic processes that constantly change. Over time, some types of bacteria may acquire or lose resistance to specific antibiotics. A major problem with standard, lab-based antibiotic resistance tests is that bacteria's sensitivity to antibiotics often differs when they are grown in a petri dish versus when they are grown in your body. This is because bacteria are not static organisms; they adapt to their environment. Propionibacterium acnes bacteria that grow in hair follicles and feed on sebum have a different metabolic profile than those grown in petri dishes and feeding on bacterial supplements. Moreover, cells regulate surface proteins, cell wall structure, and genes according to their environment, and these changes can have profound implications for their sensitivity to specific antibiotics. As a result, in the case of topical antibiotics used to treat bacterial skin conditions, there is no prior knowledge—that is, no mechanism to predict whether an antibiotic will be effective against *Propionibacterium acnes* or any other skin bacterial condition—until it is tested in a bacterial strain. For example, as shown at http: / / thescienceofacne.com / antibiotic-susceptibility-of-propionibacterium-acnes / , *Propionibacterium acnes* has been reported to be highly resistant to nitroimidazole (metronidazole) or tetracycline (lysine), but partially responsive to doxycycline (another tetracycline), and shows no resistance to ciprofloxacin but resistance to another fluoroquinolone, levofloxacin. Therefore, it is impossible to predict whether an antibiotic will work based on prior activity in other bacterial strains. Systematic development of novel antibiotics showing anti-acne activity is needed.

[0063] In this regard, DART molecules can serve as ideal drug candidates for the treatment of acne caused by Propionibacterium acnes, and additionally for the treatment of skin and skin structure infections caused by other bacteria, such as MRSA. DART is designed to contain two distinct chemical domains selected from the aforementioned different families, such as a β-lactam ring and a quinolone nucleus, or a quinolone nucleus and a nitro heterocycle, or a β-lactam ring and a nitro heterocycle, which confers two distinct mechanisms of action. This creates fewer opportunities for mutations at the two target sites in the bacteria, leading to less development of resistance to these antibiotics. Some molecules can exert additional anti-inflammatory mechanisms to reduce the host inflammatory response, further enhancing their anti-acne efficacy.

[0064] The embodiments of the various aspects disclosed herein are based on molecules designed by the inventors that can function on at least two different or distinct targets. Generally, the molecule comprises at least two different or distinct chemical domains. Each of the chemical domains binds to a distinct or distinct active site in the target cell. The chemical domains may be linked together by a third domain. As used herein, the term "chemical domain" refers to a portion of the molecule involved in the desired property. For example, a chemical domain may be a portion of the molecule involved in the binding of the molecule to a target, or a portion involved in the regulation of target activity.

[0065] In some embodiments, the first and second chemical domains independently possess antibacterial or bactericidal activity. In some embodiments, the first and second chemical domains may independently comprise an antibacterial agent. As used herein, the terms "antibacterial agent" or "antibiotic agent" are defined as compounds that have a bactericidal or bacteriostatic effect on bacteria in contact with the compound. As used herein, the term "bactericidal" is defined as having a destructive killing effect on bacteria. As used herein, the term "bacteriostatic" is defined as having an inhibitory effect on bacterial growth. Examples of antibacterial agents include, but are not limited to, macrolides or ketolides, such as erythromycin, azithromycin, clarithromycin, erythromycin, spiramycin, telithromycin, carbomycin A, josamycin, tylosin, midecamycin acetate, pyromycin, solithromycin, spiramycin, quizalofop-p-ethyl, solithromycin, spiramycin, anisomycin, pyromycin, carbomycin, and tylosin; β-lactams, including penicillins, cephalosporins, and carbapenems, such as carbapenems, imipenem, and meropenem; and monolactams, such as penicillin G, penicillin V, and methoxyfenozide. Cyclocillin, oxacillin, cloxacillin, dicloxacillin, nevcillin, ampicillin, azlocillin, amoxicillin, carbenicillin, ticarcillin, mezlocillin, piperacillin, azlocillin, temoxicillin, flucloxacillin, cephalosporins, cefepime, cefadroxil, cefamandole, cefuroxime, cefalexin, cefprozil, cefaclor, chloramphenicol, cefoxitin, cefmetazole, cefotaxime, cefazolin, ceftriaxone, cefoperazone, ceftazidime, cefixime, cefpodoxime, cefbufen, cefdinir, cefpirome, cefepime, cefadroxil, cephalosporin Thiophene, cephalexin, cefuroxime, ceftolun, ceftazidime, cefazolin oxime, cefuroxime proxetil, cefuroxime, cefepime, aztreonam, ertapenem, doripenam, and cilastatin; penicillin combinations, such as amoxicillin / clavulanic acid, ampicillin / sulbactam, piperacillin / tazobactam, and ticarcillin / clavulanate; quinolones, such as nalidixic acid, oxaquinic acid, norfloxacin, pefloxacin, enoxacin, ofloxacin, levofloxacin, ciprofloxacin, timafloxacin, lomefloxacin, fleroxacin, gapafloxacin, sparfloxacin, trovafloxacin, clindamycin, gatifloxacin, moxifloxacin, and others. Floxacin, sitafloxacin, galafloxacin, gemifloxacin, pazufloxacin, besifloxacin, eulifloxacin, prulifloxacin, sinofloxacin, pyrrolimidic acid, pipemidic acid, rosofloxacin, rufloxacin, balofloxacin, tosufloxacin, derafloxacin, nalnofloxacin; antibacterial sulfonamides and antibacterial p-aminobenzenesulfonamides, including p-aminobenzoic acid, sulfadiazine, silver sulfadiazine, sulfisoxazole, sulfamethoxazole, sulfadiazine, sulfadoxine, sulfadiazine and phthalimidethiazole, sulfamidosulfonamide, sulfacetyl, sulfadimethoxine, sulfonamide, sulfasalazine and 2,4-diaminoazobenzene-4-sulfonamide;Aminoglycosides, such as streptomycin, neomycin, kanamycin, paromomycin, gentamicin, tobramycin, amikacin, netilmicin, spectinomycin, sisomicin, dibekalin, isipamicin, dihydrostreptomycin, neomycin B, ribosomycin, abekacin, kanamycin B, dibekacin, hygromycin B, methylzolylmycin, and asmicin; tetracyclines, such as tetracycline, chlortetracycline, demethylchlortetracycline, minocycline, oxytetracycline, methacycline, doxycycline, clomocycline, and lysine. Meclocycline, penicillin, and pyrrocycline; rifamycins, such as rifampicin, rifapentine, bezoxazin, orifamycin, and rifaximin; lincosamides, such as lincomycin and clindamycin; lipopeptides, such as daptomycin; glycopeptides, such as vancomycin, tervacin, and teicoplanin; and streptotropic antibiotics, such as quinupristin and daflopristin. Ansarmycin derivatives, such as geldamicin, chloramphenicol, and rifaximin; oxazolidinones, such as linezolid, epirazolid, preszolid, radizolid, ranbezolid, sutezolid, and terdizolid; pleurotinoids, such as retamoline, tiamulin, and vorimidine; antibacterial sterols, such as fusidic acid; amide alcohols, such as chloramphenicol, chloramphenicol azidomethacin, thiamphenicol, and florfenicol; nitrofurans, such as furazolidone and nitrofurantoin; chain... Positive-positive antibiotics, such as pannamycin, quinupristin / dalfopristin, and virginiamycin; other antibacterial agents, such as arsphenamine, fosfomycin, mupirocin, acanthoxycycline, tigecycline, trimethoprim, polymyxin, bacitracin, polymyxin, colistin, metronidazole, trimethoprim-sulfamethoxazole, and fosfomycin; and antimycobacterial agents, such as chlorpheniramine, dapsone, capreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampin, rifabutin, rifapentine, and streptomycin. In some embodiments, the antibacterial agent may be selected from the group consisting of azithromycin, roxithromycin, cefuroxime, cefotaxime, cefoxitin, ceftriaxone, cephalosporins, minocycline, naflufloxacin, moxifloxacin, bexifloxacin, eulifloxacin, prulifloxacin, retamorin, metronidazole, ornidazole, and any combination thereof. In some embodiments, the antibacterial agent may be hyaluronic acid or a derivative thereof.

[0066] In some embodiments of the DART molecule, the first and second domains independently possess anti-acne activity. In some embodiments, the first and second chemical domains are independently anti-acne agents. As used herein, the term "anti-acne agent" refers to any chemical substance effective in the treatment of acne and / or related symptoms. Anti-acne agents are well known in the art, for example, by reference to U.S. Patent Publication No. 2006 / 0008538 and U.S. Patent No. 5607980, the contents of which are incorporated herein by reference. Examples of useful anti-acne agents include, but are not limited to: keratolytic agents such as salicylic acid, salicylic acid derivatives, and resorcinol; retinoids such as retinoic acid, tretinoin, adapalene, and tazarotene; sulfur-containing D- and L-amino acids, and their derivatives and salts; lipoic acid; antibiotics and antimicrobial agents such as benzoyl peroxide, triclosan, chlorhexidine gluconate, oxymethopyrone, tetracycline, 2,4,4'-trichloro-2'-hydroxydiphenyl ether, 3,4,4'-trichlorosymmetrical diphenylurea, nicotinamide, tea tree oil, rofecoxib, azelaic acid and its derivatives, phenoxyethanol, phenoxypropanol, phenoxyisopropanol, ethyl acetate, clindamycin, erythromycin, and meclocycline; sebostats such as flavonoids; and bile salts such as shark cholesterol sulfate and its derivatives, deoxycholates, and bile salts; and combinations thereof. These agents are well known and commonly used in the field of personal care.

[0067] In some embodiments, the anti-acne agent may be an antimicrobial peptide having activity against Propionibacterium acnes. Antimicrobial peptides are ubiquitous in nature and play an important role in the innate immune system of many species [Zasloff et al., 2002; and Epand et al., 1999]. Antimicrobial peptides may be naturally occurring peptides or analogs thereof, or they may be synthetic peptides. As used herein, "analyte" refers to a naturally occurring antimicrobial peptide that has been chemically modified to enhance its potency and / or reduce its toxicity. Antimicrobial peptides may be peptides known to be effective against Gram-positive bacteria. Non-limiting examples include lanthanide antibiotics, such as nisin, subtilisin, epidermin, and gallidermin; defensins; adjuvants, such as flesh fly antimicrobial peptides; cephalosporins, such as cephalosporin A, bactericides, and lepidopterans; Xenopus antimicrobial peptides; bee venom peptides; histone-rich peptides; brevinins; and combinations thereof. In addition, antimicrobial peptides active against Propionibacterium acnes have been reported, for example in U.S. Patent Publications 2005 / 0282755, 2005 / 02455452, 2005 / 0209157, and U.S. Patent No. 6255279, all of which are incorporated herein by reference. Suitable examples of antimicrobial peptides active against Propionibacterium acnes include, but are not limited to, novispirins (Hogenhaug, see above), and those antimicrobial peptides described in U.S. Patent Publication 2007 / 0265431, the contents of which are incorporated herein by reference. In some embodiments, the antimicrobial peptide may be cathilicidine and its derivatives.

[0068] In some embodiments, the antibacterial agent can be a free fatty acid (FFA) or a fatty acid derivative, or a fatty acid ester or glycerol (G) derivative of propylene glycol (PG), and any combination thereof. Examples include lauric acid, stearic acid, myristic acid, oleic acid, linoleic acid, myristoleic acid, palmitoleic acid, linolenic acid, linolenic acid, sapienic acid, various polyunsaturated fatty acids (PUFAs), propylene glycol monolaurate, glyceryl monolaurate and / or glyceryl dilaurate, propylene glycol monooleate, glyceryl monooleate and / or glyceryl dioleate, and other derivatives known in the art. Fatty acids are well-known antimicrobial agents [Kabara et al., 1972], and their activity varies with the chain length, degree of unsaturation, and number of fatty acid esters present in the propylene glycol or glycerol backbone. The primary target of FAs and their derivatives is the bacterial cell membrane, which is essentially nonspecific. Disruption of the bacterial membrane can lead to disruption of intracellular electron transport activity, oxidative phosphorylation, inhibition of specific enzyme activity, reduced cellular energy production, impaired nutrient absorption, autoxidation of degradation products, toxic peroxidation, or direct lysis of bacterial cells. Their broad-spectrum, non-specific activity makes them promising antimicrobial candidates for the treatment and prevention of antimicrobial infections caused by Gram-positive and Gram-negative bacteria that produce a wide range of skin and skin structure infections. In some embodiments, FA and its derivatives, alone or in combination with any antibiotic or a covalent conjugate of any antibiotic (e.g., DART), are effective against antibiotic-susceptible *Propionibacterium acnes* and *Propionibacterium acnes* with low responsiveness to anti-acne products containing clindamycin, doxycycline, erythromycin, or minocycline. In some embodiments, they are effective against one or more clindamycin, minocycline, erythromycin, and / or doxycycline-resistant or resistant strains of *Propionibacterium acnes*. In some embodiments, it can prevent the development of resistant forms of the pathogen.

[0069] In some embodiments, the first and second anti-acne agents in the DART or formulations disclosed herein are independently selected from the group consisting of: acetretin, adapalene, ivermectin, azelaic acid, azithromycin, benzoyl peroxide, besifloxacin, bexarotin, cefotaxime, cefoxitin, cefuroxime, cefoperazone, cefpyrrolidone, cefotaxime, clindamycin, erythromycin, etretinate, garafloxacin, ethanol. Acids, isotretinoin, lactic acid, minocycline, moxifloxacin, N-acetylcysteine, naflufloxacin, oxymethopyrone, phenoxyethanol, phenoxypropanol, prulifloxacin, pyruvic acid, radizolide (RX-1741), resorcinol, retamoline, retinoic acid, roxithromycin, salicylic acid, sitafloxacin, sodium sulfacetamide, spironolactone, sulfacetyl, sulfur, tazarotene, retinoic acid, triclosan, eulifloxacin, metronidazole, ornidazole, urea, and any combination thereof.

[0070] In some embodiments, the first and second chemical domains are independently antifungal agents. As used herein, the term "antifungal agent" refers to a substance capable of inhibiting or preventing the growth, survival, and / or reproduction of fungal cells. Preferred antifungal agents are those capable of preventing or treating fungal infections in animals or plants. Preferred antifungal agents are broad-spectrum antifungal agents. However, antifungal agents may also be specific to one or more particular species of fungi.

[0071] Examples of antifungal agents include, but are not limited to: azoles (e.g., fluconazole, isaconazole, itraconazole, ketoconazole, miconazole, clotrimazole, voriconazole, posaconazole, rivanazole, and ciclopirox ketone), polyenes (e.g., natamycin, russmycin, nystatin, and amphotericin B), echinocandins (e.g., caspofungin), praldidines (e.g., beanomicins, nicotinamides, sordarins, allylamines), triclosan, pyroxen and its olamine salt, buphenomorpholine, terbinafine, and their derivatives and analogues. Other antifungal agents include those described in the following patents: for example, International Patent Publications Nos. WO2001 / 066551, WO2002 / 090354, WO2000 / 043390, WO2010 / 032652, WO2003 / 008391, WO2004 / 018485, WO2005 / 006860, WO2003 / 086271, WO2002 / 067880, and U.S. Patent Application Publication No. 2008 / 0194661, 2008 / 0287440, 2005 / 0130940, 2010 / 0063285, 2008 / 0032994, 2006 / 0047135, 2008 / 0182885, and U.S. Patent Nos. 6,812,238, 4,588,525, 6,235,728, 6,265,584, 4,942,162, and 6,362,172, all of which are incorporated herein by reference.

[0072] 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 combinations thereof. Non-metallic salts of pyrithione, such as ethanolamine salts, deacetylated chitosan salts, and disulfide salts (commercially known as OMADINE MDS or OMDS), can also be used. The pyrithione salt can be used in any particulate form, including but not limited to crystalline forms (e.g., platelets, rods, needles, bulk crystals), round and amorphous, regular or irregular shaped particles.

[0073] In some embodiments, the hydroxypyridinethione salt is zinc hydroxypyridinethione. Zinc hydroxypyridinethione is best known for its use in treating dandruff and seborrheic dermatitis. It also has antibacterial properties and is effective against many pathogens from the genera *Streptococcus* and *Staphylococcus*. Other medical applications include the treatment of psoriasis, eczema, tinea, fungal infections, athlete's foot, dry skin, atopic dermatitis, tinea corporis, and vitiligo.

[0074] In some embodiments, the antifungal agent is an antifungal peptide. Antifungal peptides are well known in the art (see, for example, De Lucca et al., 2000). Antifungal peptides can be naturally occurring peptides or analogs thereof, or they can be synthetic peptides. As used herein, the term "analyte" refers to a naturally occurring antifungal peptide that has been chemically modified to enhance its potency and / or reduce its toxicity / side effects. Exemplary antifungal peptides may include, but are not limited to, syringostatins, syringotoxins, nicotinamides, echinocandins, pneumocadins, and aculin. Antifungal peptides include eacins, mulundocadins, cecropins, α-defensins, β-defensins, novispirins, and combinations thereof. Other antifungal peptides include the agents described in the following patents, for example, U.S. Patent No. 6,255,279 and U.S. Patent Application Publications Nos. 2005 / 0239709, 2005 / 0187151, 2005 / 0282755, and 2005 / 0245452, all of which are incorporated herein by reference.

[0075] In some embodiments, the first chemical domain is an antibacterial agent, and the second chemical domain is an anti-acne agent or an antifungal agent.

[0076] Non-limiting, the first and second chemical domains in DART can be covalently bonded to each other. Those skilled in the art will readily recognize the various functional groups within the chemical domains that can be used to covalently bond the first and second chemical domains. For example, the first chemical domain may contain functional groups selected from the group consisting of: amino groups, N-substituted amino groups, carboxyl groups, carbonyl groups, acid anhydride groups, aldehyde groups, hydroxyl groups, epoxy groups, thiols, disulfides, alkenyl groups, hydrazine groups, acylhydrazine groups, aminourea groups, aminothiourea groups, amide groups, aryl groups, ester groups, ether groups, glycidyl groups, halogen groups, hydride groups, isocyanate groups, urea groups, carbamate groups, and any combination thereof. In some embodiments, the second chemical domain includes a functional group selected from the group consisting of: amino groups, N-substituted amino groups, carboxyl groups, carbonyl groups, acid anhydride groups, aldehyde groups, hydroxyl groups, epoxy groups, thiols, disulfide groups, alkenyl groups, hydrazine groups, acylhydrazine groups, aminourea groups, aminothiourea groups, amide groups, aryl groups, ester groups, ether groups, glycidyl groups, halogen groups, hydride groups, isocyanate groups, urea groups, carbamate groups, and any combination thereof. In some embodiments, the first and second chemical domains are bonded to each other via the same functional group. In some embodiments, the first and second chemical domains are bonded to each other via different functional groups.

[0077] In some embodiments, the third domain may enhance or increase the activity of at least one chemical domain. For example, the activity of at least one chemical domain may be increased or enhanced relative to the absence of the third domain. In some embodiments, the third domain may increase or enhance the antibacterial activity of at least one chemical domain in DART. In some embodiments, the third domain may increase or enhance the anti-acne activity of at least one chemical domain in DART. In some embodiments, the third domain may increase or enhance the anti-inflammatory activity of at least one chemical domain in DART.

[0078] In some embodiments, the third domain itself is biologically active. For example, the third domain can be an active agent. In some embodiments, the third domain may have antibacterial or antifungal activity. In some embodiments, the third domain may have anti-inflammatory activity.

[0079] The third domain of DART can be a direct bond or atom (e.g., oxygen or sulfur), or a unit (e.g., NR). 1C(O), C(O)O, C(O)NH, SS, SO, SO2, SO2NH) or atomic chains (e.g., substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalenyl, arylalkynyl, heteroarylalkyl, heteroarylalenyl, heteroarylalkynyl, heterocyclic alkyl, heterocyclic alkenyl, heterocyclic alkynyl, aryl, heteroaryl, heterocyclic, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalenyl, alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalenyl, alkynylarylalkynyl, alkane Alkyl-heteroaryl, alkyl-heteroaryl-alkenyl, alkyl-heteroaryl-alkynyl, alkenyl-heteroaryl-alkenyl, alkenyl-heteroaryl-alkynyl, alkynyl-heteroaryl-alkenyl, alkynyl-heteroaryl-alkynyl, alkyl-heterocyclic alkyl, alkyl-heterocyclic alkenyl, alkyl-heterocyclic alkynyl, alkenyl-heterocyclic alkenyl, alkenyl-heterocyclic alkyl, alkynyl-heterocyclic alkenyl, alkynyl-heterocyclic alkynyl, alkylaryl, alkenylaryl, alkynyl-heterocyclic alkynyl, alkylaryl, alkenylaryl, alkyl-heteroaryl, alkenyl-heterocyclic alkynyl, wherein one or more methylene groups can be O, S, S(O), SO2, N(R) 1 )2. C(O), C(O)O, cleavable linking group, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic interruption or termination), wherein, R 1 It is a hydrogen group, acyl group, aliphatic group, or a substituted aliphatic group.

[0080] In some embodiments, the first and second chemical domains are covalently linked to each other via a third domain comprising at least one cleavable group. The cleavable group is a group that is sufficiently stable under the first set of conditions and capable of breaking to release the two parts held together by the cleavable group. In a preferred embodiment, the cleavable group breaks at least 10 times faster, preferably at least 100 times faster, under the first reference conditions (e.g., selected to simulate or represent intracellular conditions) than under the second reference conditions (e.g., selected to mimic or represent conditions in blood or serum).

[0081] Cleavable groups are sensitive to cleavage agents, such as pH, redox potential, or the presence of degrading molecules. Typically, cleavage agents are more prevalent at the sites where the molecule containing the cleavable group is required for action, or are found to be present at higher levels or with higher activity. Examples of such degrading agents include: redox agents present in cells that are selected for specific substrates or are non-substrate specific, including, for example, oxidases or reductases, or reducing agents (e.g., thiols) that can degrade redox cleavable linkages by reduction; esterases; amidases; reagents or endosomes that can create an acidic environment, such as reagents or endosomes that cause the pH to be below 5; enzymes that can hydrolyze or degrade acidic cleavable linkages by acting as generalized acids, peptidases (which can be substrate-specific), and proteases, as well as phosphatases.

[0082] Exemplary cleavable groups include, but are not limited to, redox cleavable groups (e.g., -SS- and -C(R)2-SS-, wherein R is H or a C1-C6 alkyl group, and at least one R is a C1-C6 alkyl group such as CH3 or CH2CH3); phosphate ester-based cleavable linking groups (e.g., -OP(O)(OR)-O-, -OP(S)(OR)-O-, -OP(S)(SR)-O-, -SP(O)(OR)-O-, -OP(O)(OR)-S-, -SP(O)(OR)-S-, -OP(S)(ORk)-S-, -SP(S)(OR)-O-, -OP(O)(R)-O-, -OP(S)(R)-O-, -SP(O ... -O-, -SP(S)(R)-O-, -SP(O)(R)-S-, -OP(S)(R)-S-, -OP(O)(OH)-O-, -OP(S)(OH)-O-, -OP(S)(SH)-O-, -SP(O)(OH)-O-, -OP(O)(OH)-S-, -SP(O)(OH)-S-, -OP(S)(OH)-S-, -SP(S)(OH)-O-, -OP(O)(H)-O-, -OP(S)(H)-O-, -SP(O)(H)-O-, -SP(S)(H)-O-, -SP(O)(H)-S- and -OP(S)(H)-S-, where R is an optionally substituted straight-chain or branched C1-C 10 Alkyl groups); acid cleavable groups (e.g., hydrazones, esters, and esters of amino acids such as -OC(O)-, -C=NN-); ester-based cleavable groups (e.g., -C(O)O-); peptide-based cleavable groups (e.g., groups cleaved by enzymes such as peptidases and proteases, such as –NHCHR). A C(O)NHCHR B C(O)-, where R A and RB (It is an R group consisting of two adjacent amino acids). The peptide-based cleavable group contains two or more amino acids. In some embodiments, the peptide-based cleavable group contains an amino acid sequence that is a substrate for peptidases or proteases found in or secreted by Propionibacterium acnes.

[0083] In some embodiments, the cleavable group is an acid-labile group. Typically, acid-cleavable groups are cleavable in acidic environments with a pH below about 6.5 (e.g., below about 6.5, 6.0, 5.5, 5.0, 4.5, 4.0, 3.5, 3.0) or by reagents such as enzymes that can act as generalized acids.

[0084] In some embodiments, the first and second chemical domains are covalently linked by a third domain selected from the group consisting of 11-hydroxyundecenoic acid, 1,10-decanediol, 1,3-propanediol, 1,5-pentanediol, 10-hydroxydecenoic acid, succinic acid, lactic acid, 3-hydroxypropionic acid, and any combination thereof.

[0085] In some implementations, the third structural domain may be a joint, for example, a breakable joint or a non-breakable joint.

[0086] The first chemical domain can be bonded to the second chemical domain via direct bonds or atoms (such as oxygen or sulfur), units (such as NH, C(O), C(O)O, C(O)NH, SS, SO, SO2 or SO2NH), or to a domain that connects the first and second chemical domains.

[0087] Similarly, the second chemical domain can be bonded to the first chemical domain via direct bonds or atoms (such as oxygen or sulfur), units (such as NH, C(O), C(O)O, C(O)NH, SS, SO, SO2 or SO2NH), or to a domain that connects the first and second chemical domains.

[0088] DART can be synthesized using methods known in the art. Exemplary methods for synthesizing DART are described in the Embodiments section of this document. See Embodiments 2 through 10.

[0089] In some implementations, DART may be selected from the molecules shown in Tables 1A and 1B.

[0090] Table 1A: Exemplary DART List 1

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097] Table 2A: Exemplary DART List - 2

[0098]

[0099]

[0100]

[0101] The present invention also provides formulations containing DART as an API. Various characteristics of the formulations will be described in more detail below.

[0102] Antibiotics (non-DART)

[0103] The present invention also contemplates compounds that are not DART. Therefore, the present invention also provides formulations comprising non-DART antibiotic agents, i.e., formulations comprising a non-DART antibiotic agent as an API. For example, the present invention describes the use of 8-chlorofluoroquinolone for the treatment of acne, particularly acne caused by resistant forms of Propionibacterium acnes. It is a subclass of fluoroquinolones with a chlorine-substituted C8 position. Therefore, in some embodiments, this disclosure provides formulations comprising 8-chlorofluoroquinolones as APIs. Exemplary 8-chlorofluoroquinolones include, but are not limited to, besifloxacin, sitafloxacin, and clinfloxacin. In some embodiments, the formulation comprises besifloxacin.

[0104]

[0105] Without being bound by theory, micronization of antibiotic agents (e.g., besifloxacin) can affect their biological activity. For example, micronization can enhance the biological activity of antibiotic agents or increase their retention at desired sites. Furthermore, micronization can affect the amount and stability of antibiotic agents in formulations. Additionally, micronization can allow for the optimization of the properties of formulations containing micronized besifloxacin. Therefore, without limitation, APIs (e.g., antibiotic agents) in formulations can be in the form of particles, powders, suspensions, dispersions, emulsions, liposomes, micelles, spheres, solutions, vesicles, aggregates, etc.

[0106] In some implementations, the API (e.g., but not limited to besifloxacin or DART) may be micronized, i.e., formed into granules.

[0107] Typically, the size range of micronized APIs is from about 0.2 μm to about 15 μm. In some embodiments, the size range of micronized APIs is from about 1 μm to about 10 μm. In some embodiments, the size range of micronized APIs is from about 1.5 μm to about 9 μm. In some embodiments, the size range of micronized APIs is from about 2 μm to about 8 μm.

[0108] In some implementations, the API is in particulate form and contains a surface modifier on its surface. Typically, a surface modifier is a molecule that can alter the surface of the particles under consideration (e.g., through coating) to assist in attaching the entire particle to a specific surface. Generally, surface modification does not involve changes in chemical bonding or the formation of any chemical bonds. The surface modifier is only physically associated with the particles.

[0109] Surface modifiers can be selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof. Surface modifiers can form a coating layer on the particle surface. Non-limitingly, particles can be partially or completely coated with a surface modifier.

[0110] Some non-limiting exemplary formulations of micronized antibiotic agents (e.g., besifloxacin) are described in Examples 18-20 and are shown in Table 18.

[0111] In some embodiments, the formulation may be a spray formulation. Exemplary, non-limiting spray formulations are described in Example 23 and Table 19. In some embodiments, the formulation may be in the form of a facial cleanser. Exemplary, non-limiting facial cleansers are described in Example 24 and Table 20. In some embodiments, the formulation may be in the form of a soap bar. Exemplary, non-limiting soap bar formulations are described in Example 25 and Table 21. In some embodiments, the formulation may be in the form of a shower gel. Exemplary, non-limiting shower gel formulations are described in Example 26 and Table 22. In some embodiments, the formulation may be in the form of a lotion. Exemplary, non-limiting lotion formulations are described in Example 27 and Table 23.

[0112] Surfactants are known to solubilize hydrophobic substances by reducing interfacial tension. Therefore, in some embodiments, antibiotic formulations may be solubilized with surfactants prior to the formation of the formulation. In addition to surfactants, cosolvents or cosurfactants may also help solubilize poorly soluble water compounds by increasing the wettability of hydrophobic molecules or reducing their interfacial tension. Some exemplary surfactants and cosurfactants may include, but are not limited to, sodium lauryl sulfate, Tween 80, Tween 20, Span 20, and any combination thereof. Exemplary cosolvents for solubilizing antibiotic formulations (e.g., besifloxacin) may include propylene glycol monocaprylate and diethylene glycol monoethyl ether. Other tested surfactants, cosurfactants, and cosolvents for solubilizing antibiotic formulations are described elsewhere in this disclosure. It is not intended to be theoretically binding, but solubilizing antibiotic formulations (e.g., besifloxacin) may provide formulations that comply with FDA formulation guidelines and restrictions on inactive excipients or ingredients. Non-limiting exemplary formulations containing a solubilizing API (e.g., an antibiotic formulation such as besifloxacin) are described in Examples 28, 31, and 33 and are shown in Tables 24, 27, and 30-32.

[0113] Preparing drug-loaded (suspension) gels using conventional methods often exposes the drug to a wide range of pH conditions, which in some cases can lead to drug dissolution followed by redeposition. This dissolution-reprecipitation phenomenon often results in changes to the original particle size, impurity distribution, or crystal structure. To circumvent this problem, the inventors have used the method of the present invention to prepare various suspension-loaded drug formulations. Thus, in some embodiments, the formulation is in the form of a suspension gel with negligible or minimal drug dissolution-reprecipitation. In suspension drug formulations, API particles are dispersed in a carrier medium (e.g., but not limited to glycerol) and processed to obtain the desired formulation. Exemplary suspension gel formulations are described in Examples 24, 26, 28, 29, 30, 31, and 32 and are shown in Tables 25, 27, 29, 33, 34, 37, and 38. Exemplary suspension-loaded drug cream formulations are described in Examples 30 and 32 and Tables 26 and 28.

[0114] In addition to the various components, the formulation may also contain one or more viscosity modifiers. In some embodiments, the viscosity modifier is a polymer. Exemplary polymeric viscosity modifiers include, but are not limited to, carbopol, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxyethyl cellulose, and sodium hyaluronate. In some other embodiments, the viscosity modifier is a non-polymeric viscosity modifier or gelling agent. Other exemplary viscosity modifiers are described elsewhere in this disclosure. Exemplary formulations containing various viscosity modifiers are described in Example 33 and Tables 29-32.

[0115] According to published literature, some physical and / or chemical interactions may exist between carbopol and fluoroquinolones. Therefore, it may be necessary to prepare formulations that do not contain carbopol or similar polymers to avoid any incompatibility issues during product storage. Thus, in some embodiments, the formulations are substantially free of viscosity modifiers. Exemplary formulations that are substantially free of viscosity modifiers are described in Example 32 and Table 28.

[0116] In some embodiments, the API may be coated with molecules selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof. Without limitation, the API may be partially or completely coated with coating molecules. Exemplary formulations comprising coated or uncoated APIs are described in Example 46 and Tables 52 and 53.

[0117] It is worth noting that the various formulation characteristics discussed in more detail below apply to formulations containing antibiotic agents (such as besifloxacin) as described herein.

[0118] combination

[0119] In some embodiments, the formulation comprises two or more antibiotic agents. For example, the formulation may comprise two or more different anti-acne agents. In some embodiments, the formulation comprises a single 8-chlorofluoroquinolone or a combination thereof with another anti-acne agent. Exemplary 8-chlorofluoroquinolones include, but are not limited to, besifloxacin, sitafloxacin, and clinfloxacin. In some embodiments, the formulation comprises besifloxacin. In some embodiments, the formulation comprises besifloxacin and adapalene.

[0120] Non-limitingly, two or more antibiotic agents may be in the same or different forms. For example, the first and second antibiotic agents may be independently micronized, suspended, or solubilized for use in an API. Therefore, in some embodiments, the first and second antibiotic agents are micronized. In some embodiments, the first antibiotic agent is micronized and the second antibiotic agent is solubilized. In some embodiments, the first antibiotic agent is micronized and the second antibiotic agent is suspended in the formulation. In some embodiments, the first antibiotic agent is solubilized and the second antibiotic agent is micronized. In some embodiments, both the first and second antibiotic agents are solubilized. In some embodiments, the first antibiotic agent is solubilized and the second antibiotic agent is suspended. In some embodiments, the first antibiotic agent is suspended and the second antibiotic agent is micronized. In some embodiments, the first antibiotic agent is suspended and the second antibiotic agent is solubilized. In some embodiments, both the first and second antibiotic agents are suspended.

[0121] In some embodiments, the formulation comprises besifloxacin and adapalene, wherein besifloxacin is solubilized and adapalene is suspended. In other embodiments, the formulation comprises besifloxacin and adapalene, wherein both besifloxacin and adapalene are solubilized. Exemplary formulations comprising both besifloxacin and adapalene are described in Examples 18, 23-27, and 31 and in Tables 18-23 and 27.

[0122] It is worth noting that the various formulation characteristics discussed in more detail below apply to formulations containing two or more antibiotic agents as described herein.

[0123] Features applicable to DART, non-DART, and composite APIs

[0124] Furthermore, as described at http: / / thescienceofacne.com / antibiotic-susceptibility-of-propionibacterium-acnes / , 'A second major limitation in acne treatment is the non-uniform distribution of antibiotics across different tissues of the body. Many antibiotics do not effectively accumulate in hair follicles and / or sebaceous glands, and therefore cannot effectively reach the bacteria that cause acne. Even if bacteria are highly sensitive to a particular antibiotic in laboratory-based tests, if that antibiotic cannot reach the site of infection in sufficient concentration, then it is not an effective treatment. As a result, there is a major difference in the effectiveness of oral and topical antibiotics used in acne treatment.' This situation extends to all bacterial diseases of the skin. There is a need to develop unique, optimal topical formulations, which will be described later.

[0125] Skin, such as microcracks, sweat or secretory pores, and hair follicles, can serve as reservoirs for drug carriers of specific sizes. Funnel-shaped delivery can be used to enhance the potency of active agents, such as antifungal and antibacterial agents. Drug carriers can enhance the delivery of active agents to sebaceous follicles and also exhibit fusogeneity towards the cell walls / membranes of lipophilic microorganisms. This allows the drug carrier to remain on the skin, subsequently releasing DART or antibacterial agents slowly and continuously. Exemplary drug carriers include, but are not limited to, microparticles, nanoparticles, vesicles, liposomes, emulsions, microspheres, and solutions.

[0126] In addition to the API (e.g., DART and / or other antimicrobial agents), the drug carrier may further comprise one or more other components. For example, the drug carrier may further comprise compounds selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof. The API (e.g., DART or other antimicrobial agents) may be present in the core of the drug carrier, and the other components may form a coating layer around the core. Without limitation, the coating layer may be a functional or non-functional coating layer. A functional coating layer is a coating layer that imparts one or more desired properties to the drug carrier, such as enhancing targeting or retention at the site of action, increasing API activity, or possessing desired activity itself.

[0127] In some embodiments, DART and / or other antibacterial agents may be formed into granules. In addition to APIs (such as DART and / or other antibacterial agents), the granules may further comprise compounds selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, and any combination thereof. APIs (such as DART or other antibacterial agents) may be present in the core of the granules, and other components may form a coating layer on the core.

[0128] In some embodiments, the particles contain a surface modifier located on their surface. Typically, a surface modifier is a molecule that can alter the surface of the particle in question (e.g., through coating) to assist in attaching the entire particle to a specific surface. Generally, surface modification does not involve changes in chemical bonding or the formation of any chemical bonds. The surface modifier is only physically associated with the particle.

[0129] Surface modifiers can be selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof. Surface modifiers can form a coating layer on the particle surface. Non-limitingly, particles can be partially or completely coated with a surface modifier.

[0130] In some embodiments, the drug delivery systems and formulations disclosed herein may further comprise an active agent, i.e., an active agent other than DART and / or antibacterial agents. As used herein, the term "active agent" refers to a compound or composition having a specific desired activity. For example, an active agent may be a therapeutic compound. Without limitation, the active agent may be selected from the group consisting of: small organic or inorganic molecules, saccharins, oligosaccharides, polysaccharides, peptides; proteins, peptide analogs and derivatives, peptide mimics, nucleic acids, nucleic acid analogs and derivatives, antibodies, antigen-binding fragments of antibodies, lipids, extracts made from biological materials, naturally occurring or synthetic compositions, and any combination thereof.

[0131] In some embodiments, the active agent may be selected from the group consisting of: antifungal agents, antibacterial agents, antimicrobial agents, antiacne agents, antioxidants, cooling agents, soothing agents, wound healing agents, anti-inflammatory agents, penetration enhancers, permeation enhancers, antioxidants, anti-aging agents, anti-wrinkle agents, skin whitening or bleaching agents, ultraviolet (UV) absorbers or scatterers, skin depigmenting agents, regenerating agents, scar healing agents, dyes or coloring agents, deodorants, fragrances, keratolytic agents, and any combination thereof. In some embodiments, the active agent may be a keratolytic agent.

[0132] In some embodiments, the active agent is an anti-inflammatory agent. As used herein, the term "anti-inflammatory agent" refers to a compound (including its analogues, derivatives, prodrugs, and pharmaceutically acceptable salts) that can be used to treat inflammation or inflammation-related diseases or disorders. Exemplary anti-inflammatory agents include, but are not limited to, known steroidal anti-inflammatory drugs and nonsteroidal anti-inflammatory drugs (NSAIDs). Exemplary steroidal anti-inflammatory agents include, but are not limited to: 21-acetoxygestrinone, aclomethasone, alpha-pregnane, ancinonide, beclomethasone, betamethasone, budesonide, cloprednisolone, clobetasol, clobetansone, clocotropin, cloprednisolone, corticosteroids, cortisone, cortisol, divazoline, dexamethasone, dexamethasone, difluralasone, diflucortisone, difluprednisolone, glycyrrhetinic acid, fluzacrocin, fluclochloride, flumethasone, flunisolone, flunicolone, and fluocinolone acetonide. Fluocinolone acetonide, fluocinonide, flucobutyl ester, fluocinolone, fluocinolone, fluperazine, fluprednisolone acetate, fluprednisolone acetate, fluprednisolone, fluticasone propionate, formococcal, halcinonide, halobetasol propionate, halometasone, haloprednisolone acetate, hydrocortisone, hydrocortisone, chlorteprednisolone etopolate, horseprednidone, methylhydroxysone, methylprednisolone, methylprednisolone, mometasone furoate Prednisolone, prednisone, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednisolone valerate, prednisolone, limesole, tecortisone, triamcinolone, triamcinolone hexacetonide, derivatives of the above substances, and mixtures thereof. Exemplary nonsteroidal anti-inflammatory agents include, but are not limited to, COX inhibitors (COX-1 or COX nonspecific inhibitors) and selective COX-2 inhibitors. Exemplary COX inhibitors include, but are not limited to: salicylic acid derivatives, such as aspirin, sodium salicylate, magnesium trisalicylate choline, salicylates / esters, diflunisal, sulfasalazine, and oxalazine; p-aminophenol derivatives, such as acetaminophen; indole and indoleacetic acid, such as indole-methyl and sulindac; heteroarylaceous acids, such as tometidine, diclofenac, and ketorolac; arylpropionic acids, such as ibuprofen, naproxen, flurbiprofen, ketoprofen, fenofoprofen, and oxaprazin; anthranilic acid (methanoic acid) derivatives, such as mefenamic acid and meloxicam; enolic acids, such as cocoa derivatives (pyroxicam, meloxicam); ketones, such as nabumetone; their derivatives and mixtures thereof. Exemplary COX-2 inhibitors include, but are not limited to: diaryl-substituted furanones, such as rofecoxib; diaryl-substituted pyrazoles, such as celecoxib; indoleacetic acids, such as etodoxacin; and sulfonylanilines, such as nimesulide; their derivatives and mixtures thereof.

[0133] In some embodiments, the active agent is an anti-aging agent. As used herein, the term "anti-aging agent" refers to a compound or composition that inhibits or reduces signs of aging, such as wrinkles, fine lines, and other manifestations of photodamage. Examples of anti-aging agents include, but are not limited to: flavonoids, such as quercetin, hesperidin, quercetin, rutin, citrus flavonoids, and epicatechin; CoQ10; inorganic sunscreens, such as titanium dioxide and zinc oxide; organic sunscreens, such as octyl methyl cinnamate and its derivatives; retinoids; vitamins, such as vitamin E, vitamin A, vitamin C (ascorbic acid), vitamin B, and their derivatives (e.g., vitamin E acetate, vitamin C palmitate, etc.); antioxidants including α-hydroxy acids, such as glycolic acid, citric acid, lactic acid, malic acid, mandelic acid, ascorbic acid, α-hydroxybutyric acid, α-hydroxyisobutyric acid, α-hydroxyisohexanoic acid, atrrolactic acid, α-hydroxyisovaleric acid, ethyl pyruvate, galacturonic acid, glucopehtonic acid, and glucopheptonoic acid. 1,4-lactone, gluconic acid, gluconic acid lactone, glucuronic acid, glucurronolactone, glycolic acid, isopropyl pyruvate, methyl pyruvate, mucoic acid, pyruvate, glycolic acid, glycolic acid 1,4-lactone, tartaric acid, and malonic acid; β-hydroxy acids, such as β-hydroxybutyric acid, β-phenyllactic acid, and β-phenylpyruvate; plant extracts, such as green tea, soybean, milk thistle, seaweed, aloe vera, angelica, bitter orange, coffee, coptis, grapefruit, hoellen, honeysuckle, coix seed, lithospermum, mulberry, white peony root, kudzu root, rice, safflower, and mixtures thereof.

[0134] In some embodiments, the active agent is an ultraviolet (UV) absorber or scattering agent. UV absorbers include, for example, benzoic acid-based UV absorbers, such as para-aminobenzoic acid (hereinafter referred to as PABA), PABA monoglyceride, N,N-dipropoxy PABA ethyl ester, N,N-diethoxy PABA ethyl ester, N,N-dimethyl PABA ethyl ester, N,N-dimethyl PABA butyl ester, and N,N-dimethyl PABA methyl ester; anthranilic acid-based UV absorbers, such as N-acetyl-anthranilic acid menthol ester; and salicylic acid-based UV absorbers, such as salicylic acid methyl ester. Amyl salicylate, menthyl salicylate, high-menthol salicylate, octyl salicylate, phenyl salicylate, benzyl salicylate, p-isopropanol phenyl salicylate, etc.; UV absorbers in the cinnamic acid system, such as octyl cinnamate, ethyl 4-isopropylcinnamate, methyl 2,5-diisopropylcinnamate, ethyl 2,4-diisopropylcinnamate, methyl 2,4-diisopropylcinnamate, propyl p-methoxycinnamate, isopropyl p-methoxycinnamate, isoamyl p-methoxycinnamate, octyl p-methoxycinnamate (p-methoxycinnamate 2-... Ethylhexyl ester), 2-ethoxyethyl p-methoxycinnamate, cyclohexyl p-methoxycinnamate, ethyl α-cyano-β-phenylcinnamate, 2-ethylhexyl α-cyano-β-phenylcinnamate, glyceryl mono-2-ethylhexanoyl-di-p-methoxycinnamate, bis(trimethylsiloxane)silylisopentyltrimethoxycinnamate methyl ester, etc.; 3-(4'-methylbenzylene)-d,l-camphor; 3-benzylene-d,l-camphor; uric acid, ethyl uric acid; 2-phenyl-5-methylbenzoxazole; 2, 2'-Hydroxy-5-methylphenylbenzotriazole; 2-(2'-hydroxy-5'-tert-octylphenyl)benzotriazole; 2-(2'-hydroxy-5'-methylphenyl)benzotriazole; dibenzaladine; dianisoylmethane; 4-methoxy-4'-tert-butyldibenzoylmethane; 5-(3,3-dimethyl-2-norbornyl)-3-pentane-2-one; dimorpholinopyridazinone; and combinations thereof. Ultraviolet scattering agents include, for example, powders, such as titanium dioxide, particulate titanium dioxide, zinc oxide, particulate zinc oxide, iron oxide, particulate iron oxide, and cerium oxide.

[0135] In some embodiments, the active agent is an anti-wrinkle agent, such as a dermatological anti-wrinkle agent. Anti-wrinkle agents include, but are not limited to, flavonoids, such as quercetin, hesperidin, quercetin, rutin, citrus flavonoids, and epicatechin; CoQ10; vitamin C; including C2-C. 30Hydroxy acids, including α-hydroxy acids such as glycolic acid, lactic acid, 2-hydroxybutyric acid, malic acid, citric acid, tartaric acid, α-hydroxyacetic acid, hydroxyoctanoic acid, etc.; β-hydroxy acids, including salicylic acid and polyhydroxy acids such as gluconolactone (G4); and mixtures of these acids. Further, anti-wrinkle agents include retinoic acid and gamma-linolenic acid.

[0136] In some embodiments, the active agent is a skin brightener or bleaching agent. Skin brighteners and bleaching agents include: hydrogen peroxide, zinc peroxide, sodium peroxide, hydroquinone, 4-isopropylcatechol, hydroquinone monobenzyl ether, kojic acid; lactic acid; ascorbic acid and its derivatives, such as magnesium ascorbate phosphate; arbutin; and licorice. Active ingredients for sun-free tanning include: dihydroxyacetone (DHA); glyceraldehyde; tyrosine and tyrosine derivatives, such as malyltyrosine, tyrosine glucosinate, and ethyltyrosine; DOPA phosphate, indole and its derivatives; and mixtures thereof. Other skin whitening agents include glycosamines, such as glucosamine, N-acetylglucosamine, glucosamine sulfate, mannosamine, N-acetylmmannosamine, galactosamine, N-acetylgalactosamine, their isomers (e.g., stereoisomers), and their salts (e.g., hydrochlorides); and N-acyl amino acid compounds, such as N-acylphenylalanine, N-acyltyrosine, their isomers (including their D and L isomers), salts, derivatives, and mixtures thereof. A suitable example of an N-acyl amino acid is N-undecenoyl-L-phenylalanine.

[0137] In some embodiments, the active agent is a skin depigmenting agent. Examples of suitable depigmenting agents include, but are not limited to: soybean extract; soy isoflavones; retinoids, such as retinol; kojic acid; kojic acid dipalmitate; hydroquinone; arbutin; transexamic acid; vitamins, such as niacin and vitamin C; azelaic acid; linolenic acid and linolenic acid; placertia; licorice; and extracts such as chamomile and green tea; as well as their salts and prodrugs.

[0138] In some embodiments, the active agent is an antioxidant. As used herein, the term "antioxidant" refers to any molecule capable of slowing down, reducing, inhibiting, or preventing the oxidation of other molecules. Examples of antioxidants include, but are not limited to, hydrophilic antioxidants, lipophilic antioxidants, and mixtures thereof. Non-limiting examples of hydrophilic antioxidants include chelating agents (e.g., metal chelating agents), such as ethylenediaminetetraacetic acid (EDTA), citrate, ethylene glycol tetraacetic acid (EGTA), 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), diethylenetriaminepentaacetic acid (DTPA), 2,3-dimercapto-1-propanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), α-lipoic acid, salicylaldehyde isonicotinamide hydrazone (SIH), hexylthioethylamine hydrochloride (HTA), deferoxamine, their salts, and mixtures thereof. Other hydrophilic antioxidants include ascorbic acid (vitamin C), cysteine, glutathione, dihydrolipoic acid, 2-mercaptoethanesulfonic acid, 2-mercaptobenzimidazole sulfonic acid, 6-hydroxy-2,5,7,8-tetramethylchromo-2-carboxylic acid, sodium metabisulfite, their salts, and mixtures thereof. Non-limiting examples of lipophilic antioxidants include: vitamin E isomers, such as α-, β-, γ-, and δ-tocopherols and α-, β-, γ-, and δ-tocotrienols; polyphenols, such as 2-tert-butyl-4-methylphenol, 2-tert-butyl-5-methylphenol, and 2-tert-butyl-6-methylphenol; butylated hydroxyanisoles (BHAs), such as 2-tert-butyl-4-hydroxyanisole and 3-tert-butyl-4-hydroxyanisole; butylated hydroxytoluene (BHT); tert-butylhydroquinone (TBHQ); ascorbate palmitate; n-propyl gallate; and their salts; and mixtures of the above substances. Those skilled in the art will understand that antioxidants can be classified into primary antioxidants, secondary antioxidants, or metal chelators based on their mechanisms of action. Primary antioxidants quench free radicals typically originating from oxidative pathways, while secondary antioxidants exert their effects by decomposing peroxides, which are active intermediates in oxidative pathways. Metal chelating agents function by isolating trace amounts of metals that promote free radical formation. In some embodiments, the antioxidant is resveratrol.

[0139] In some embodiments, the active agent is a wound healing agent. As used herein, the term "wound healing agent" refers to an active agent that is effective in promoting the healing process of natural wounds over days, weeks, or months. Exemplary wound healing agents include, but are not limited to, protein growth factors, vascular endothelial growth factors, antiproliferative agents, antimicrobial agents, and anti-inflammatory agents.

[0140] In some embodiments, the active agent is a soothing agent. As used herein, the term "soothing agent" refers to a molecule that, for example, helps reduce skin and / or scalp discomfort by relieving the sensation of itching. Exemplary soothing agents include, but are not limited to, aloe vera, avocado oil, green tea extract, hops extract, chamomile extract, gelatinous oatmeal, calamine, cucumber extract, sodium palmate, sodium palm kernel oil, shea butter, peppermint leaf oil, sericin, pyridoxine (a form of vitamin B6), retinyl palmitate and / or other forms of vitamin A, tocopherol acetate and / or other forms of vitamin E, lauryl laurate, hyaluronic acid, aloe vera leaf juice powder, eutectic kale (a type of berry) fruit extract, riboflavin (vitamin B2), thiamine hydrochloride and / or other forms of vitamin B1 and / or any combination thereof.

[0141] In some embodiments, the active agent is a cooling agent. As used herein, the term "cooling agent" refers to a molecule that provides a cooling sensation in application. Some exemplary cooling agents include, but are not limited to: WS-3, WS-23, menthol, 3-substituted-P-menthane, N-substituted-P-menthane-3-carboxamide, isoprene, 3-(1-menthoxy)propane-1,2-diol, 3-(1-menthoxy)-2-methylpropane-1,2-diol, p-menthane-2,3-diol, p-menthane-3,8-diol, 6-isopropyl-9-methyl-1,4-dioxane[4,5]decane-2-methanol, menthyl succinate and its alkaline earth metal salts, trimethylcyclohexanol, N-ethyl-2-isopropyl 5-Methylcyclohexaneformamide, Japanese peppermint oil, peppermint oil, menthone, menthone glyceryl ketal, menthyl lactate, 3-(1-menthoxy)ethyl-1-ol, 3-(1-menthoxy)prop-1-ol, 3-(1-menthoxy)but-1-ol, 1-menthylacetic acid N-ethylamide, 1-menthyl-4-hydroxyvalerate, 1-menthyl-3-hydroxybutyrate, N,2,3-trimethyl-2-(1-methylethyl)-butanamide, n-ethyl-t-2-c-6-nonadienamide, N,N-dimethylmenthylsuccinamide, menthylpyrrolidone carboxylic acid ester, etc.

[0142] In some embodiments, the active agent is a colorant. As used herein, the term "colorant" refers to any substance that can be used to produce a desired color. Typically, such colorants are approved for human consumption by appropriate government agencies and / or laws (such as the Food and Drug Administration (FDA) / Federal Food, Drug and Cosmetic Act (FD&C) or similar bodies in the European Union). For example, a colorant can be a food-grade dye or a lake. A "dye" is a water-soluble compound that is available as a powder, granules, liquid, or other special-purpose form. A "lake" is a non-water-soluble form of a dye. Exemplary colorants include, but are not limited to, FD&C Blue 1 (Brilliant Blue), FD&C Blue 2 (Indigo), FD&C Green 3 (Fixed Green), FD&C Red 3 (Erythrosine), FD&C Red 40 (Allura Red), FD&C Yellow 5 (Tartrazine), FD&C Yellow 6 (Sunset Yellow), annatto extract, anthocyanins, chokeberry / red berries, beet juice, beet powder, β-carotene, β-apo-8-carotenal, blackcurrant, caramel, canthaxanthin, maltose, pharmaceutical charcoal, carmine, carmine / β-carotene, and carmine blue. The preferred colorants according to the present invention are FD&C Blue 1 (Brilliant Blue), FD&C Blue 2 (Indigo), FD&C Green 3 (Fixed Green), FD&C Red 3 (Erythrosine), FD&C Red 40 (Allura Red), FD&C Yellow 5 (Tartrazine), FD&C Yellow 6 (Sunset Yellow), and any combination thereof.

[0143] In some embodiments, the active agent is a fragrance. Exemplary fragrances include, but are not limited to: 2,4-dimethyl-3-cyclohexene-1-carboxaldehyde; isocyclocitral; menthone; isomenthone; ROMASCONE ® (2,2-Dimethyl-6-methylene-1-cyclohexanecarboxylate methyl ester); Nerolidin; Terpineol; Dihydroterpineol; Terpene acetate; Dihydroterpene acetate; Dipentene; Eucalyptol; Hexanoate / ester; Rose ether; Perycollle ® ((S)-1,8-p-menthadien-7-ol); 1-p-menthene-4-ol; (1RS,3RS,4SR)-3-p-menthyl acetate; (1R,2S,4R)-4,6,6-trimethyl-bicyclo[3,1,1]hept-2-ol; DOREMOX ®(Tetrahydro-4-methyl-2-phenyl-2H-pyran); Cyclohexyl acetate; Cycloalkyl acetate; Fructalate (1,4-cyclohexanediethyl dicarboxylate); KOUMALACTONE ® ((3ARS,6SR,7ASR)-perhydro-3,6-dimethyl-benzo[B]furan-2-one); Natactone ((6R)-perhydro-3,6-dimethyl-benzo[B]furan-2-one); 2,4,6-trimethyl-4-phenyl-1,3-dioxane; 2,4,6-trimethyl-3-cyclohexen-1-carboxaldehyde; (E)-3-methyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)-4-penten-2-ol; (1'R,E)-2-ethyl-4-(2',2',3'-trimethyl-3'-cyclopenten-1'-yl)-2-buten-1-ol; POLYSANTOL ® ((1'R,E)-3,3-dimethyl-5-(2',2',3'-trimethyl-3'-cyclopenten-1'-yl)-4-penten-2-ol); 2-heptaylcyclopentanone (fleuramone); Paradisone ® ((1R)-cis-3-oxo-2-pentyl-1-cyclopentanemethyl acetate); Veloutone (2,2,5-trimethyl-5-pentyl-1-cyclopentanone); NIRVANOL ® (3,3-Dimethyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)-4-penten-2-ol); 3-Methyl-5-(2,2,3-trimethyl-3-cyclopenten-1-yl)-2-pentanol; damascones; neobutterone ® (1-(5,5-dimethyl-1-cyclohexen-1-yl)-4-penten-1-one); nectalactone ((1'R)-2-[2-(4'-methyl-3'-cyclohexen-1'-yl)propyl]cyclopentanone); α-ionone; β-ionone; damascenone; DYNASCONE ® (A mixture of 1-(5,5-dimethyl-1-cyclohexen-1-yl)-4-penten-1-one and 1-(3,3-dimethyl-1-cyclohexen-1-yl)-4-penten-1-one); DORINONE ® β (1-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2-buten-1-one); ROMANDOLIDE ® ((1S,1'R)-[1-(3',3'-dimethyl-1'-cyclohexyl)ethoxycarbonyl]methylpropionate); 2-tert-butyl-1-cyclohexyl acetate; LIMBANOL ®(1-(2,2,3,6-tetramethylcyclohexyl)-3-hexanol); trans-1-(2,2,6-trimethyl-1-cyclohexyl)-3-hexanol; (E)-3-methyl-4-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3-buten-2-one; terpenoid isobutyrate; LORYSIA ® (4-(1,1-dimethylethyl)-1-cyclohexylacetate); 8-methoxy-1-p-menthene; HELVETOLIDE ® ((1S,1'R)-2-[1-(3',3'-dimethyl-1'-cyclohexyl)ethoxy]-2-methylpropylpropionate); p-tert-butylcyclohexanone; menthenethiol; 1-methyl-4-(4-methyl-3-pentenyl)-3-cyclohexen-1-carboxaldehyde; allyl cyclohexylpropionate; cyclohexyl salicylate; methyl cypressone; verdylate; vetyverol; vetyverone; 1-(octahydro-2,3,8,8) -Tetramethyl-2-naphthyl)-1-ethyl ketone; (5RS,9RS,10SR)-2,6,9,10-tetramethyl-1-oxaspiro[4.5]dec-3,6-diene and (5RS,9SR,10RS)-isomer; 6-ethyl-2,10,10-trimethyl-1-oxaspiro[4.5]dec-3,6-diene; 1,2,3,5,6,7-hexahydro-1,1,2,3,3-pentamethyl-4-indanone; HIVERAL ® (A mixture of 3-(3,3-dimethyl-5-indanyl)propionaldehyde and 3-(1,1-dimethyl-5-indanyl)propionaldehyde); Rhubofix ® (3',4-Dimethyl-tricyclo[6.2.1.0(2,7)]undec-4-ene-9-spiro-2'-ethylene oxide); 9 / 10-Ethylene-3-oxatricyclo[6.2.1.0(2,7)]undecane; POLYWOOD ® (Fully hydrogenated 5,5,8A-trimethyl-2-naphthylacetate); octalynol; CETALOX ®(dodecano-3a,6,6,9a-tetramethyl-naphtho[2,1-b]furan); tricyclo[5.2.1.0(2,6)]dec-3-en-8-yl acetate and tricyclo[5.2.1.0(2,6)]dec-4-en-8-yl acetate and tricyclo[5.2.1.0(2,6)]dec-3-en-8-yl propionate and tricyclo[5.2.1.0(2,6)]dec-4-en-8-yl propionate; camphor; borneol; isoborneol acetate; 8-isopropyl-6-methyl-bicyclo[2.2.2]oct-5-en-2-carboxaldehyde; camphopinene; cedarwood methyl ether (8-methoxy-2,6,6,8-tetramethyl-tricyclo[5.3.1.0(1,5)]undecane); cedarwoodene; cedarwood alcohol; cedarwood alcohol; FLOREX ® A mixture of (9-ethylene-3-oxatricyclo[6.2.1.0(2,7)]undec-4-one and 10-ethylene-3-oxatricyclo[6.2.1.0(2,7)]undec-4-one; 3-methoxy-7,7-dimethyl-10-methylene-bicyclo[4.3.1]decane; CEDROXYDE ® (trimethyl-13-oxabicyclo[10.1.0]tetadeca-4,8-diene); Ambrettolide LG ((E)-9-hexadecane-16-lactone); HABANOLIDE ® (Pentadecanolide); Muscone (3-methyl-(4 / 5)-cyclopentadecanolide); Muscone; Exaltolide ® (Decadecyl lactone); EXALTONE ® (Cyclopentadecanone); (1-ethoxyethoxy)cyclododecane; Astrotone; Lilial ® ; pine oil, etc.

[0144] In some embodiments, the active agent is an antifungal agent. Exemplary antifungal agents are described elsewhere herein. As used herein, the term "fungus" includes a variety of spore-bearing nucleated organisms that do not contain chlorophyll. Examples include yeasts, molds, mildews, rust fungi, and mushrooms. Examples of fungi include, but are not limited to, Aspergillus fumigatus (…). Aspergillus fumigates Aspergillus flavus Aspergillus flavus Aspergillus nidus ( ) Aspergillus nidulans Candida albicans ( Candida albicans ), Candida glabrata ( Candida glabrata ), Gilead Candida ( Candida guilliermondii ), Candida krusei ( Candida krusei ), Candida Portugueseum ( Candida lusitaniae ), Candida parapsilosis ( Candida parapsilosis), Tropical Candida ( Candida tropicalis Cryptococcus neoformans ( Cryptococcus neoformans ), Oriental Isaac yeast ( Issatchenkia orientalis ), Coccidioides Coccidioides Paracoccidioides ( Paracoccidioides Histoplasma capsulatum ( ) Histoplasma ), Blastomyces ( Blastomyces ), Trichophyton rubrum ( Trichophyton rubrum ) and Neurospora crassa ( Neurospora crassa In some embodiments, the fungus is of the genus Malassezia (e.g., Malassezia furfur). M. furfur Malassezia (thick-skinned Malassezia) M. pachydermatis ), Spherical Malassezia ( M. globosa ), Restrictive Maselachus ( M. restricta ), Slofi Malassezia ( M. slooffiae ), synaxyl Malassezia ( M. sympodialis ), Nanamaraspermum ( M. nana ), Malassezia dawa ( M. yamatoensis ), Malassezia cutanea ( M. dermatis ) and Malassezia abductus ( M. obtuse In one embodiment, the fungus is Trichophyton rubrum (…). Trichophyton rubrum ).

[0145] In some embodiments, the active agent is an antibacterial agent. Exemplary antibacterial agents are described elsewhere in this disclosure.

[0146] In some embodiments, the active agent is an anti-scarring agent. As used herein, the term "anti-scarring agent" refers to any agent that inhibits fibrosis or scar formation. Useful anti-scarring agents can inhibit one or more aspects of the fibrotic process. For example, in some embodiments, anti-scarring agents inhibit inflammation; inhibit the production or release of collagen in cells; and / or act as anti-infective or antifungal agents. In some embodiments, the anti-scarring agent is selected from the group consisting of (-)-arbuscular induration 6. The device, wherein the anti-scarring agent is selected from: angiogenesis inhibitors, 5-HT inhibitors, β1 integrin antagonists, β-tubulin inhibitors, bisphosphonate compounds selected from risedronate and its analogues or derivatives, enzyme generation blockers in hepatitis C, bone mineralization promoters, Bruton's tyrosine kinase inhibitors, calcineurin inhibitors, calcium channel blockers, calmodulin kinase II inhibitors, caspase 3 inhibitors, cathepsin B inhibitors, cathepsin K inhibitors, cathepsin L inhibitors, CB1 / CB2 receptor agonists, CC chemokine receptor antagonists, CD40 antagonists, cell cycle inhibitors, chemokine receptor antagonists, chymotrypsin inhibitors, coagulation factors, collagenase antagonists, cual integrin inhibitors, CXCR antagonists, cyclic GMP agonists, cyclin-dependent kinase inhibitors, cyclooxygenase 1 inhibitors, and dopamine D2 receptor antagonists. DHFR inhibitors, diuretics, DNA alkylating agents, DNA methylation inhibitors, DNA methylation promoters, DNA synthesis inhibitors, DNA topoisomerase inhibitors, dopamine antagonists, farnesyltransferase inhibitors, farnexyltransferase inhibitors, fibrinogen antagonists, G protein agonists, glycosylation inhibitors, heat shock protein 90 antagonists, histamine receptor antagonists, histone deacetylase inhibitors, histone deacetylase inhibitors, JAK2 inhibitors, JAK3 enzyme inhibitors, JNK inhibitors, kinase inhibitors, kinase inhibitors, kinase antagonists, leukotriene inhibitors and antagonists, lysyl hydrolase inhibitors, MAP kinase inhibitors, matrix metalloproteinase inhibitors, microtubule inhibitors, microtubule inhibitors, muscarinic receptor inhibitors, neurokinin antagonists, nitric oxide agonists, nitric oxide synthase inhibitors, NO synthase inhibitors, norepinephrine reuptake inhibitors, NSAIDs, p38 MAP kinase inhibitors, palmitoyl protein thioesterase inhibitors, PDGF receptor kinase inhibitors, peptidyl glycine α-hydroxylation monooxygenase inhibitors, peptidyl prolyl cis / trans isomerase inhibitors, peptidyl prolyl cis / trans isomerase inhibitors, peroxisome proliferator-activated receptor (PPAR) agonists, pesticides, phosphatase inhibitors, phosphodiesterase inhibitors, PKC inhibitors, platelet-activating factor antagonists, platelet aggregation inhibitors, polymorphonuclear neutrophil inhibitors,Prolyl hydroxylase inhibitors, prostaglandin inhibitors, protein synthesis inhibitors, protein tyrosine kinase inhibitors, purine receptor P2X antagonists, pyruvate dehydrogenase activators, Raf kinase inhibitors, RAR / RXT antagonists, reducing agents, retinoic acid receptor antagonists, selective serotonin reuptake inhibitors, serine protease inhibitors, serotonin receptor inhibitors, abscisic acid dehydrogenase inhibitors, sodium channel inhibitors, steroids, steroids, matrix lysozyme inhibitors, superoxide anion generators, TACE inhibitors, telomerase inhibitors, TGFβ inhibitors, thromboxane A2 receptor inhibitors, TNF-α antagonists, Toll-like receptor inhibitors Trypsin inhibitors, tubulin antagonists, tumor necrosis factor antagonists, tyrosine kinase inhibitors, VEGF inhibitors, vitamin D receptor agonists, ampicillin sodium, acetylcholinesterase inhibitors, actin polymerization and stabilization promoters, adenylate cyclase agonists, ALK-5 receptor antagonists, alpha-adrenergic receptor antagonists, androgen inhibitors, anesthetic compounds, angiotensin II receptor agonists, antibiotics (selected from the group consisting of apigenin, anticoagulants, antiemetics, anti-inflammatory compounds, antimetabolites, and antitumor agents), antimicrobial agents, antitumor agents, antioxidants, antiproliferative agents, antipsychotic compounds, and anticonvulsants. Antispasmodics, antithrombotic agents, antiviral agents, apoptosis activators, apoptosis antagonists, aromatase inhibitors, AXOR12 agonists, elastase inhibitors, e1F-2a inhibitors, elongation factor-1α inhibitors, endothelial growth factor antagonists, endothelial growth factor receptor kinase inhibitors, endotoxin antagonists, epoch-binding agents and tubulin, estrogen agonists, estrogen receptor antagonists, FGF inhibitors, FGF receptor kinase inhibitors, FLT-3 kinase inhibitors, FXR antagonists, HMGCoA reductase inhibitors, ICAM inhibitors, IL, IL-2 inhibitors Immunosuppressants, type III receptor tyrosine kinase inhibitors, inosine monophosphate inhibitors, interleukin antagonists, intracellular calcium flux inhibitors, intracellular calcium influx inhibitors, irreversible enzyme inhibitors of methionine aminopeptidase type 2, isoenzyme-selective delta protein kinase C inhibitors, MCP-CCR2 inhibitors, MEK1 / MEK2 inhibitors, MIF inhibitors, mTOR inhibitors, mTOR kinase inhibitors, NFκB inhibitors, ornithine decarboxylase inhibitors, S-adenosyl-L-homocysteine ​​hydrolase inhibitors, SDF-1 antagonists, SRC inhibitors, Syk kinase inhibitors, α-glucosidase inhibitors Antagonists, and immunomodulators selected from Bay 11-7085, and IRAK antagonists, ICE, imidazolidinedione hydrochloride, protein kinase B inhibitors, protein kinase C stimulators, purine nucleoside analogs, puromycin, reversible inhibitors of ErbB1 and ErbB2, ribonucleoside triphosphate reductase inhibitors, and any combination thereof. In some embodiments, the anti-scarring agent may be selected from: ZD-6474, AP-23573, synthadotin, S-0885, april, ixaprone, IDN-5390, SB-2723005, ABT-518, cobustatin, anecoxetine acetate, SB-715992, sirolimus esterified, adalimumab, erucic acid phosphocholine, alphastatin, etanercept, humicade, gefitinib, isosorbide dinitrate, etc. Retinoic acid, rhizobacterin, clobetasol propionate, homoharringtonine, trichomycin A, brefidobacterin A, carotenoids, doralastatin, cerivastatin, jasplakinolide, atrazine A, pirfenidone, vinorelbine, 17-DMAG, tacrolimus, clotipreno, juglone, prednisolone, puromycin, 3-BAABE, cladribine, mannose-6-phosphate, 5-azacytidine, Ly333531 (rubusta) and simvastatin.

[0147] In some implementations, the active agent is a skin regeneration agent. Some skin regeneration agents can also act as anti-scarring agents.

[0148] In some embodiments, the drug carrier comprises an additional anti-acne agent. In some embodiments, the additional anti-acne agent may be selected from the group consisting of: acitretin, adapalene, retinoic acid, α-hydroxy acids or β-hydroxy acids, antibiotics, antimicrobial peptides, antibiotic agents, azelaic acid, benzoyl peroxide, bexarotine, bile salts, biofilm inhibitors, clindamycin, erythromycin, etretinate, glycolic acid, isotretinoin, keratolytic agents, lactic acid, lipoic acid, N-acetylcysteine, natural anti-acne agents, oxymetazoline, phenoxyethanol, phenoxypropanol, pyruvate, resorcinol, retinoic acid, retinoids, salicylic acid, sebostats, sodium sulfacetamide, spironolactone, sulfur, sulfur-containing D-amino acids or L-amino acids, tazarotene, tea tree oil, retinoic acid, triclosan, urea, and any combination thereof.

[0149] The drug delivery systems disclosed herein may contain any amount of API (e.g., DART or other reagents). For example, the drug delivery system may contain from about 0.01% to about 99% (w / w) of API. For example, the particles may contain between about 0.01% and about 20% (w / w) of API. In some embodiments, the API constitutes more than 1% (w / w), more than 5% (w / w), more than 10% (w / w), more than 15% (w / w), more than 20% (w / w), more than 25% (w / w), more than 30% (w / w), more than 35% (w / w), more than 40% (w / w), more than 45% (w / w), more than 50% (w / w), more than 55% (w / w), more than 60% (w / w), more than 65% (w / w), more than 70% (w / w), more than 75% (w / w), more than 80% (w / w), more than 85% (w / w), more than 90% (w / w), or more than 95% (w / w) of the total weight of the drug carrier. In some embodiments, the API content in the drug carrier may be from about 75% to about 97% (w / w). In other embodiments, the API content in the drug carrier may be from about 3% to about 25% (w / w).

[0150] The lipids used in the drug delivery systems or formulations disclosed herein may be selected from the group consisting of: fatty acids; fatty alcohols; glycerides (e.g., monoglycerides; diglycerides and triglycerides); phospholipids; glycerophospholipids; sphingolipids; sterol esters; isopentenol esters; glycolipids; polyketides; and any combination thereof. In some embodiments, the lipids may be selected from the group consisting of: 1,3-propanediol dioctanoate / didecanoate; 10-undecenoic acid; 1-triacosanol; 1-heptacosanol; 1-nonacosanol; 2-ethylhexanol; androstenedane; arachidonic acid; arachidonic acid; arachidonic acid; behenic acid; behenol; Capmul MCM C10; decanoic acid; decanol; octanol; caprylic acid; saturated fatty alcohols C12-C18 caprylate / capric ester; caprylic / capric triglyceride; caprylic / capric triglyceride; ceramide phosphorylcholine (sphingomyelin, SPH); ceramide phosphorylethanolamine (sphingomyelin, Cer-PE); ceramide phosphorylglycerol; waxy acid; waxy acid; wax alcohol; cetearyl alcohol; cetyl ether-10; cetyl alcohol; cholene; cholesterol; cis-11-eicosenoic acid; cis-11-octadecenoic acid; cis-13-docosahexaenoic acid; cluytyl alcohol; coenzyme Q10 (CoQ10); dihydroxy-gamma-linolenic acid; docosahexaenoic acid; lecithin; eicosapentaenoic acid; eicosaenoic acid; trans-oleic acid; trans-octadecenoyllinolenic acid alcohol); trans-octadecenelinolenic acid; trans-oleic acid; erucic acid; mustard; estradiol; ethylene glycol distearate (EGDS); gestigmic acid; gestigmol; glyceryl distearate (type I) EP (Precirol ATO 5); tricaprylate / capric acid ester; tricaprylate / capric acid ester (CAPTEX) ®355 EP / NF); Capmul MCM C8 EP; Triglyceride; Tricaprylyl; Tricaprylyl / Capric / Lauryl; Tricaprylyl / Tricapric; Tripalmitate; Heptanoic acid; Heptanoic acid; Heptanoic acid; Heptanoic acid; Heptanoic acid; Isostearic acid; Isostearyl alcohol; Lacceroic acid; Lauric acid; Laureth; Tetracosanoic acid; Tetracosanoic acid; Linolenic acid; Linolenic acid; Linolenic acid; Heptanoic acid; Mead; Bee acid; Bee alcohol; Linaloic acid; Linalool; Myricyl alcohol alcohol); myristic acid; myristoleic acid; myristol; neodecanoic acid; neoheptanoic acid; neononanoic acid; nervonic acid; nonacosanic acid; nonacosanol; nonacosanic acid; nonacosanic acid; oleic acid; oleyl alcohol; palmitic acid; palmitoleic acid; palmitole alcohol; nonanoic acid; nonanol; pentadecanoic acid; pentadecanoic acid; phosphatidic acid (phosphatidyl ester, PA); phosphatidylcholine (lecithin, PC); phosphatidylethanolamine (cephalin, PE); phosphatidylinositol (PI); phosphatidylinositol diphosphate (PIP2); phosphatidylinositol phosphate (PIP); phosphatidylinositol triphosphate (PIP3); phosphatidylserine (PS); polyglycerol-6-distearate; pregnane; propylene glycol didecanoate; propylene glycol dioctyldecanoate; propylene glycol dioctyldecanoate; triterpenoid acid; recinoleaic acid; ricinoleol; sapienic acid (acid); soybean lecithin; stearic acid; octadecanoic acid; stearyl alcohol; tris(oleic acid); tridecanoic acid; tridecanoic acid; trioleic acid glyceride; undecanoic acid; undecenoic acid; undecanoic acid; isoleic acid; α-linolenic acid; γ-linolenic acid; fatty acid salts of the following substances, including 10-undecenoic acid, adapalene, arachidic acid, arachidonic acid, behenic acid, butyric acid, capric acid, caprylic acid, ceric acid, cis-11-eicosenoic acid, cis-11-octadecenoic acid, cis-13-docosahexaenoic acid, docosahexaenoic acid, eicosapentaenoic acid. Ethylenediol, erucic acid, icosanoic acid, heptadecanoic acid, heptadecanoic acid, isostearic acid, lauric acid, tetracosanoic acid, trans-linolenic acid, linoleic acid, linoleic acid, myristic acid, myristoleic acid, neodecanoic acid, neoheptanoic acid, neononanoic acid, nonadecanoic acid, oleic acid, palmitic acid, palmitoleic acid, nonanoic acid, pentadecanoic acid, ricinoleic acid (such as zinc ricinoleate), hexadecenoic acid, stearic acid, tridecanoic acid, tridecanoic acid, undecenoic acid, undecanoic acid, isoleic acid, valeric acid, α-linolenic acid, or γ-linolenic acid; paraffin; and any combination thereof. In some embodiments, the lipid may be a fatty acid containing 11 or fewer carbons. For example, the fatty acid may contain 6, 7, 8, 9, 10, or 11 carbons.

[0151] Not wanting to be bound by theory, it is believed that fatty acid salts can be used in particles to enhance antibacterial activity (e.g., anti-acne activity) and provide stability in compositions containing the drug carrier. Therefore, in some embodiments, the lipid is a fatty acid salt. Without limitation, the fatty acid salt may be selected from the group consisting of zinc salts, sodium salts, potassium salts, lithium salts, ammonium salts, copper salts, calcium salts, magnesium salts, strontium salts, manganese salts, and combinations thereof. The drug carrier may contain any amount of lipid component. For example, the drug carrier may contain between about 0.01% and about 99% (w / w) of lipid component. In some embodiments, the lipid component accounts for more than 0.1% (w / w), more than 0.5% (w / w), more than 1% (w / w), more than 2% (w / w), more than 3% (w / w), more than 4% (w / w), more than 5% (w / w), more than 6% (w / w), more than 7% (w / w), more than 8% (w / w), more than 9% (w / w), more than 10% (w / w), more than 11% (w / w), and more than 12% of the total weight of the drug load. The percentages of lipid components in drug delivery systems are typically 2%–25% (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), greater 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). Generally, the lipid component content in drug delivery systems ranges from approximately 2% to 25% (w / w).

[0152] The ratio of the surfactant (e.g., DART or other antibacterial agents) to the total lipid component of the coating layer can be any desired ratio. For example, the ratio of surfactant to total lipid component can range from about 100:1 to about 1:100. In some embodiments, the ratio of surfactant to total lipid component can range from about 75:1 to about 1:75, about 50:1 to about 1:50, about 25:1 to about 1:25, about 20:1 to about 1:20, about 15:1 to about 1:15, about 5:1 to about 1:5, or about 25:1 to about 1:5. In some embodiments, the ratio of surfactant to total lipid component is about 30:1, about 25:1, about 20:1, about 15:1, about 10:1, about 5:1, or about 1:1. This ratio can be based on weight, mass, or molar amount.

[0153] The thickness of the coating layer can range from nanometers to millimeters. For example, the thickness of the coating layer can range from about 1 nm to about 5000 nm, about 5 nm to about 2500 nm, about 10 nm to about 2000 nm, about 50 nm to about 1500 nm, about 20 nm to about 1000 nm, about 1 nm to about 1000 nm, about 1 nm to about 500 nm, about 1 nm to about 250 nm, about 1 nm to about 200 nm, about 1 nm to about 150 nm, about 1 nm to about 100 nm, about 2 nm to about 50 nm, or about 5 nm to about 25 nm.

[0154] In some embodiments, the drug carrier may contain two or more lipids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or more than 10), that is, the carrier may contain a first lipid and a second lipid. For example, the coating layer may contain a second lipid that is different from the first ester.

[0155] The exemplary proteins used in the drug delivery systems or formulations disclosed herein may include, but are not limited to: actin, albumin, amaranth protein, ammonium hydrolyzed animal protein, animal protein, barley protein, Brazil nut protein, casein, collagen, hydrolyzed collagen, conchiolin, corn protein, cottonseed protein, elastin, extensin, silk fibroin, fibronectin, fish protein, cod protein, gelatin, gluten, glycoprotein, hazelnut protein, hemoglobin, hemp seed protein, honey protein, hydrolyzed actin, hydrolyzed amaranth protein, hydrolyzed animal protein, hydrolyzed barley protein, hydrolyzed Brazil nut protein, hydrolyzed conchiolin, hydrolyzed corn protein, hydrolyzed cottonseed protein, hydrolyzed elastin, hydrolyzed extensin, hydrolyzed silk fibroin, hydrolyzed fibronectin, hydrolyzed fish protein, hydrolyzed cod protein, hydrolyzed cod protein, hydrolyzed gelatin, hydrolyzed hair keratin, hydrolyzed hazelnut, hydrolyzed hazelnut egg Hydrolyzed hemoglobin, hydrolyzed hemp seed protein, hydrolyzed honey protein, hydrolyzed keratin, hydrolyzed lupin protein, hydrolyzed Eurasian maple protein, hydrolyzed milk protein, hydrolyzed oat protein, hydrolyzed pea protein, hydrolyzed potato protein, hydrolyzed reticulin, hydrolyzed royal jelly protein, hydrolyzed sericin, hydrolyzed serum protein, hydrolyzed sesame protein, hydrolyzed soy protein, hydrolyzed soy milk protein, hydrolyzed spinal cord protein, hydrolyzed sponge protein, hydrolyzed almond protein, hydrolyzed plant protein, hydrolyzed wheat gluten, hydrolyzed wheat protein, hydrolyzed whey protein, hydrolyzed yeast protein, hydrolyzed yogurt protein, hydrolyzed zein, integrin, hydrolyzed jojoba protein (HP), keratin, lupin protein, Eurasian maple protein, MEA hydrolyzed collagen, MEA hydrolyzed silk, milk protein, myosin, oat protein, pea protein, polylysine, potato protein, reticulin, quaternized rice protein (Rice Quat), royal jelly protein, sericin, serum protein, sesame protein, silk powder, sodium hydrolyzed casein, soy protein, soybean peptides, soy milk protein, spinal cord protein, sponge protein, almond protein, plant protein, wheat gluten, whey protein, yeast protein, yogurt protein, corn protein, and zinc hydrolyzed collagen.

[0156] In some implementations, the protein is albumin. Albumin can be naturally occurring albumin, albumin-associated proteins, or variants thereof, such as natural or engineered variants. Variants include polymorphisms, fragments (e.g., domains and subdomains, fragments, and / or fusion proteins). Albumin can contain albumin protein sequences obtained from any source. Many proteins are known to exist in the albumin family. Therefore, albumin can contain an albumin sequence derived from one of the following serum albumin sources: African clawed frog (e.g., see Swissprot registry number P08759-1), cattle (e.g., see Swissprot registry number P02769-1), cat (e.g., see Swissprot registry number P49064-1), chicken (e.g., see Swissprot registry number P19121-1), chicken ovalbumin (e.g., see Swissprot registry number P01012-1), cobra ALB (e.g., see Swissprot registry number Q91134-1), dog (e.g., see Swissprot registry number P49822-1), donkey (e.g., see S... Swissprot Registry No. QSXLE4-1), European water frog (e.g., see Swissprot Registry No. Q9YGH6-1), schistosome (e.g., see Swissprot Registry Nos. AAL08579 and Q95VB7-1), long-clawed gerbil (e.g., see Swissprot Registry Nos. O35090-1 and JC5838), goat (e.g., see Swissprot Registry No. B3VHM9-1, available as trade number A2514 or A4164 from Sigma), guinea pig (e.g., see Swissprot Registry No. Q6WDN9-1), hamster (see DeMarco et al., 2007, International...). Journal for Parasitology, 37(11): 1201-1208), horse (e.g. see Swissprot registry P35747-1), human (e.g. see Swissprot registry P02768-1), Australian lungfish (e.g. see Swissprot registry P83517), rhesus macaque (e.g. see Swissprot registry Q28522-), mouse (e.g. see Swissprot registry P07724-1), North American bullfrog (e.g. see Swissprot registry P21847-1), pig (e.g. see Swissprot registry P08835-1), pigeon (e.g., Khan et al., 2002, 1112, J. Biol).As defined in Macromol, 30(3-4), 171-8, rabbit (see, for example, Swissprot registry number P49065-1), rat (see, for example, Swissprot registry number P02770-1), salamander (see, for example, Swissprot registry number Q8UW05-1), salmon ALB1 (see, for example, Swissprot registry number P21848-1), salmon ALB2 (see, for example, Swissprot registry number Q03156-1), and sea lamprey (see, for example, Swissprot registry number P49065-1). Albumins include those from sheep (e.g., see Swissprot registry number P14639-1), Sumatran orangutans (e.g., see Swissprot registry number Q5NVH5-1), lizards (e.g., see Swissprot registry number Q8JIA9-1), turkey ovalbumin (e.g., see Swissprot registry number O73860-1), and Xenopus laevis (e.g., see Swissprot registry number Q6D.I95-1), and the albumins include variants and fragments thereof as defined herein. Many naturally occurring albumins are known to have mutant forms. Several are described in Peters (1996, All About Albumin: Biochemistry, Genetics and Medical Applications, Academic Press, Inc., San Diego, California, pp. 170-181), the contents of which are incorporated herein by reference. The term albumin also includes albumin variants, such as genetically engineered forms, mutant forms, and fragments, having one or more binding sites that are similar to unique binding sites for one or more albumins as defined above. Similar binding sites contemplated in the context of this invention are structures capable of competing with each other to bind to identical ligand structures. In one embodiment, albumin is bovine serum albumin, ovalbumin, hydrolyzed whey protein, or lactalbumin, including variants and fragments thereof. In one embodiment, the protein is ovalbumin.

[0157] The drug carrier may contain between about 0.01% and about 99% (w / w) of protein. In some embodiments, the protein component accounts for more than 0.1% (w / w), more than 0.5% (w / w), more than 1% (w / w), more than 2% (w / w), more than 3% (w / w), more than 4% (w / w), more than 5% (w / w), more than 6% (w / w), more than 7% (w / w), more than 8% (w / w), more than 9% (w / w), more than 10% (w / w), more than 11% (w / w), and more than 1% (w / w) of the total weight of the drug carrier. 2% (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), greater 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). Typically, the protein component content in the drug carrier ranges from approximately 1%-25% (w / w), approximately 0.1%-10% (w / w), approximately 0.5%-5% (w / w), or approximately 1%-1.5% (w / w).

[0158] The ratio of the active agent (e.g., DART or other antibacterial agents) to the protein component can be any desired ratio. For example, the ratio of the active agent to the protein component can range from about 100:1 to about 1:100. In some embodiments, the ratio of the active agent to the protein component can range from about 100:1 to about 1:1, about 90:1 to about 10:1, about 85:1 to about 15:1, about 80:1 to about 25:1, or 75:1 to about 50:1. In some embodiments, the ratio of the active agent to the protein component is about 75:1. This ratio can be based on weight, mass, or molar amount.

[0159] Generally, any cationic molecule can be used in the drug delivery systems or formulations disclosed herein. As used herein, the term "cationic molecule" refers to a molecule carrying a net positive charge. In some embodiments, the cationic molecule is a polyamine. Exemplary cationic molecules include, but are not limited to, putrescine (butane-1,4-diamine), cadaverine (pentane-1,5-diamine), spermidine, spermine, cyclen (1,4,7,10-tetrachlorohexacyclododecane), cyclopamine (1,4,8,11-tetraazacyclotetradecane), linear polyethyleneimine (poly(iminoethylene)), norsinine, p-phenylenediamine (1,4-diaminobenzene), diethylenetriamine (N-(2-aminoethyl)-1,2-ethylenediamine), thermal spermine, tris(2-aminoethyl)amine, hexamethylenediamine, β-lysine (3,6-diaminohexanoic acid), m-phenylenediamine (1,3-diaminobenzene), diaminopropane (1,2-diaminopropane), ethylenediamine dihydroiodide, and polyamine D 400 (polyoxyalkyleneamine D 400).

[0160] The drug carrier may contain about 0.01% to about 99% (w / w) of cationic molecules. In some embodiments, the cationic molecules account for more than 0.1% (w / w), more than 0.5% (w / w), more than 1% (w / w), more than 2% (w / w), more than 3% (w / w), more than 4% (w / w), more than 5% (w / w), more than 6% (w / w), more than 7% (w / w), more than 8% (w / w), more than 9% (w / w), more than 10% (w / w), more than 11% (w / w), and more than 1% (w / w) of the total weight of the drug carrier. 2% (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), greater 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). Typically, the content of cationic molecules in the drug carrier ranges from approximately 1%-25% (w / w), approximately 0.1%-10% (w / w), approximately 0.5%-5% (w / w), or approximately 1%-1.5% (w / w).

[0161] The ratio of the active agent (e.g., DART or other antibacterial agents) to the protein component can be any desired ratio. For example, the ratio of the active agent to the protein component can range from about 100:1 to about 1:100. In some embodiments, the ratio of the active agent to the protein component can range from about 100:1 to about 1:1, about 90:1 to about 10:1, about 85:1 to about 15:1, about 80:1 to about 25:1, or 75:1 to about 50:1. In some embodiments, the ratio of the active agent to the protein component is about 75:1. This ratio can be based on weight, mass, or molar amount.

[0162] Generally, any carbohydrate molecule may be used in the drug delivery systems or formulations disclosed herein. In some embodiments, the carbohydrate is a polysaccharide. Exemplary polysaccharides include: cellulose derivatives, such as hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and carboxymethyl cellulose; glycosaminoglycans, such as hyaluronic acid, chondroitin sulfate, chitosan, and deacetylated chitosan; starch derivatives, such as starch / hydroxyethyl starch; agarose; and alginates / esters, and combinations thereof. In some embodiments, the carbohydrate may be selected from the group consisting of deacetylated chitosan and its derivatives, alginates / esters and their derivatives, and amylopectin and its derivatives.

[0163] The drug carrier may contain about 0.01% to about 99% (w / w) of carbohydrates. In some embodiments, the carbohydrates constitute more than 0.1% (w / w), more than 0.5% (w / w), more than 1% (w / w), more than 2% (w / w), more than 3% (w / w), more than 4% (w / w), more than 5% (w / w), more than 6% (w / w), more than 7% (w / w), more than 8% (w / w), more than 9% (w / w), more than 10% (w / w), more than 11% (w / w), and more than 1% (w / w) of the total weight of the drug carrier. 2% (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), greater 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). Typically, the carbohydrate content in the drug delivery system ranges from approximately 1%-25% (w / w), approximately 0.1%-10% (w / w), approximately 0.5%-5% (w / w), or approximately 1%-1.5% (w / w).

[0164] The ratio of active agent (e.g., DART or other antibacterial agents) to carbohydrates can be any desired ratio. For example, the range of the active agent to carbohydrate ratio can be from about 100:1 to about 1:100. In some embodiments, the range of the active agent to carbohydrate ratio can be 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 active agent to carbohydrate ratio is about 75:1. This ratio can be based on weight, mass, or molar amount.

[0165] In some embodiments, the drug carrier further comprises an excipient. In some embodiments, the excipient is a wetting agent. Without limitation, the wetting agent may be selected from alkyl sulfates, such as sodium lauryl sulfate, sodium stearyl sulfate, sodium oleyl sulfate, and sodium hexadecyl sulfate; alkyl aryl sulfonates, such as sodium dodecylbenzene sulfonate; and dialkyl sulfosuccinates, such as sodium bis(2-ethylhexyl)sulfosuccinate; sodium lauryl sulfate is most preferred. Other examples of pharmaceutically acceptable wetting agents include benzyl ethoxymonochloride, hexadecylpyridine chloride, sodium docusate, poloxamer, polysorbate, and dehydrated sorbitol esters.

[0166] In some embodiments, the excipient is a stabilizer, such as a surface stabilizer. Suitable surface stabilizers are preferably selected from known organic and inorganic pharmaceutical excipients. Such excipients include various polymers, low molecular weight oligomers, natural products, and surfactants with high and low hydrophilic-lipophilic balance (HLB) values. Preferred surface stabilizers include nonionic and ionic surfactants. Two or more surface stabilizers can be used in combination. Representative examples of surface stabilizers include sodium docusate, hexadecylpyridine chloride, gelatin, casein, lecithin (phospholipids), dextran, glycerol, gum arabic, cholesterol, astragalus gum, stearic acid, benzalkonium chloride, calcium stearate, glyceryl monostearate, cetearyl alcohol, polysitropyl emulsion wax, sorbitan esters, polyoxyethylene alkyl ethers (e.g., polyethylene glycol ethers, such as polysitropyl 1000), polyoxyethylene castor oil derivatives, and polyoxyethylene sorbitan fatty acid esters (e.g., commercially available Tween). ® For example, Twain 20 ® And Twain 80 ® (ICI Specialty Chemicals), polyethylene glycol (e.g., Carbowaxs 3350) ® and 1450 ® And Capop 934 ®(UnionCarbide), dodecyltrimethylammonium bromide, polyoxyethylene stearate, colloidal silica, phosphates / esters, sodium dodecyl sulfate, calcium carboxymethyl cellulose, hydroxypropyl cellulose (e.g., HPC, HPC-SL and HPC-L), hydroxypropyl methyl cellulose (HPMC), sodium carboxymethyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose phthalate, amorphous cellulose, magnesium aluminum silicate, triethanolamine, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), 4-(1,1,3,3-tetramethylbutyl)-phenol polymers formed with ethylene oxide and formaldehyde (also known as tyloxacillin, superione and Triton), poloxamer (e.g., poloxamer F68) ® and F108 ® Block copolymers of ethylene oxide and propylene oxide), poloxamine (e.g., Tetronic 908) ® Also known as poloxamine 908 ® Tetrafunctional block copolymers obtained by the sequential addition of propylene oxide and ethylene oxide to ethylenediamine (BASF Wyandotte, Parsippany, NJ), charged phospholipids (e.g., dimyristoyl phosphatidylglycerol, sulfosuccinate (DOSS)), Tetronic 1508 ® (T-1508) (BASF Wyandotte), sodium dialkyl sulfosuccinate (e.g., Aerosol OT) ® American Cyanamid, a sodium sulfosuccinate dioctyl ester, and Duponol P ® (Sodium lauryl sulfate (DuPont)), Tritons X200 ® (alkyl aryl polyether sulfonates (Rohm and Haas)), Crodestas F-L10 ® (A mixture of sucrose stearate and sucrose distearate (Croda Inc.)), p-Isononylphenoxy poly(glycidyl) (also known as Olin-IOG) ® Or Surfactant 10-G ® (Olin Chemicals, Stamford, CT)), Crodestas SL-40 ®(Croda, Inc.), decyl-N-methylglucosamide, n-decyl β-D-glucopyranoside, n-decyl β-D-maltopyranoside, n-dodecyl β-D-glucopyranoside, n-dodecyl β-D-maltopyranoside, heptayl-N-methylglucosamide, n-heptyl β-D-glucopyranoside, n-heptyl β-D-thioglucopyranoside, n-hexyl β-D-glucopyranoside, nonanoyl-N-methylglucosamide, n-nonyl β-D-glucopyranoside, octyl-N-methylglucosamide, n-octyl β-D-glucopyranoside, octyl β-D-thioglucopyranoside, etc. Most of these surface stabilizers are known pharmaceutical excipients and are described in detail in the Handbook of Pharmaceutical Excipients, jointly published by the American Pharmaceutical Association and the British Pharmaceutical Association (Pharmaceutical Press, 1986), the contents of which are incorporated herein by reference. In one embodiment, the excipient is sodium docusate.

[0167] Generally, the average diameter of the drug carrier is from about 5 nm to about 20,000 nm. In some embodiments, the average diameter of the drug carrier is from about 5 nm to about 5,000 nm. In some embodiments, the average diameter of the drug carrier is from about 50 nm to about 2,500 nm. In some embodiments, the average diameter of the drug carrier is from about 100 nm to about 2,000 nm. In some embodiments, the average diameter of the drug carrier is from about 150 nm to about 1,700 nm. In some embodiments, the average diameter of the drug carrier is from about 200 nm to about 1,500 nm. In some embodiments, the average diameter of the drug carrier is about 260 nm. In one embodiment, the average diameter of the drug carrier is from about 30 nm to about 150 nm. In some embodiments, the average diameter of the drug carrier is from about 100 nm to about 1,000 nm, from about 200 nm to about 800 nm, from about 200 nm to about 700 nm, or from about 300 nm to about 700 nm.

[0168] Generally speaking, the drug delivery systems disclosed herein can be in any shape or form, such as spherical, rod-shaped, elliptical, cylindrical, capsule-shaped, or disc-shaped.

[0169] In some embodiments, the drug carrier may be micrometer-sized, with a size of about 1 μm to about 1000 μm. In some embodiments, the drug carrier may be nanometer-sized, with a size of about 0.1 nm to about 1000 nm. In some embodiments, the drug carrier is a micrometer particle or a nanoparticle. As used herein, the term "micrometer particle" refers to a particle with a size of about 1 μm to about 1000 μm. As used herein, the term "nanoparticle" refers to a particle with a size of about 0.1 nm to about 1000 nm.

[0170] Those skilled in the art will understand that particles typically exhibit a size distribution around a specified “size.” Unless otherwise stated, the terms “drug carrier size” and “particle size” as used herein refer to the size distribution pattern of the drug carrier or particle, i.e., the most frequently occurring value in the size distribution. Methods for measuring drug carrier or particle size are well known to those skilled in the art, such as dynamic light scattering (e.g., optical correlation spectroscopy, laser diffraction, low-angle laser light scattering (LALLS), and medium-angle laser light scattering (MALLS)), light masking methods (e.g., Coulter analysis), or other techniques (e.g., rheology, and optical microscopy or electron microscopy).

[0171] In some embodiments, the drug carrier may be substantially spherical. "Substantially spherical" means that the ratio of the longest to the shortest vertical axis of the drug carrier's cross-section is less than or equal to about 1.5. For a substantially spherical shape, a line of symmetry is not necessary. Furthermore, the drug carrier may have surface textures, such as lines, depressions, or protrusions very small relative to the overall size of the drug carrier, and still remain substantially spherical. In some embodiments, the length ratio between the longest and shortest axes of the drug carrier is less than or equal to about 1.5, less than or equal to about 1.45, 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 1.20, less than or equal to about 1.15, or less than or equal to about 1.1. To avoid being bound by theory, the surface contact of the substantially spherical drug carrier is minimized, which minimizes undesirable aggregation of the drug carrier during storage. Many crystals or sheets have flat surfaces that allow for large surface contact areas, which can aggregate through ionic or nonionic interactions. The spherical nature of the sphere allows for contact with a much smaller area.

[0172] In some embodiments, the drug carriers have substantially the same particle size. Drug carriers with a broad size distribution (comprising both relatively large and relatively small drug carriers) allow smaller drug carriers to fill the gaps between them, thereby creating new contact surfaces. A broad size distribution can lead to larger spherical shapes by creating numerous contact opportunities for aggregation. The drug carriers described herein are within a narrow size distribution, thereby minimizing the chance of contact aggregation. A “narrow size distribution” refers to a particle size distribution where the ratio of the 90th percentile volume diameter to the 10th percentile volume diameter of small spherical particles is less than or equal to 5. In some embodiments, the ratio of the volume diameter of the 90th percentile to the volume diameter of the 10th percentile of the microsphere is less than or equal to 4.5, less than or equal to 4, less than or equal to 3.5, less than or equal to 3, less than or equal to 2.5, less than or equal to 2, less than or equal to 1.5, less than or equal to 1.45, less than or equal to 1.40, less than or equal to 1.35, less than or equal to 1.3, less than or greater than or equal to 1.25, less than or equal to 1.20, less than or equal to 1.15, or less than or equal to 1.1.

[0173] Geometric standard deviation (GSD) can also be used to indicate a narrow size distribution. GSD calculation involves determining the effective cutoff diameter (ECD) that is cumulatively less than 15.9% and 84.1%. GSD is equal to the square root of the ratio of ECD less than 84.17% to ECD less than 15.9%. A narrow size distribution is indicated by GSD < 2.5. In some embodiments, GSD is less than 2, less than 1.75, or less than 1.5. In one embodiment, GSD is less than 1.8.

[0174] Although drug carriers have been discussed from the perspective of coated particles, there are at least eight types of drug carriers that can be formulated with an active agent and one or more additional components. Different types of drug carriers can be of the following types: (1) a drug carrier comprising a core formed of an active agent, wherein the core absorbs / adsorbs additional components, or the additional components form one or more coating layers on the core of the drug carrier; (2) a drug carrier comprising an overall homogeneous mixture of an active agent and additional components; (3) a drug carrier comprising a core containing an overall homogeneous mixture of an active agent and additional components, wherein the additional components form one or more coating layers on the core of the drug carrier; (4) a drug carrier comprising a core formed of additional components, wherein the active ingredient forms one or more coating layers on the core of the drug carrier; (5) a drug carrier comprising a core containing an overall homogeneous mixture of an active agent and additional components, wherein the active agent forms one or more coating layers on the core of the drug carrier. (6) A drug carrier comprising a core, the core being a material other than an active agent and additional components, and a mixture of the active agent and additional components forming one or more coating layers on the drug carrier core; (7) A drug carrier comprising a core, the core containing a generally homogeneous mixture of the active agent and additional components, and a mixture of the active agent and additional components forming one or more coating layers on the drug carrier core; (8) A liposome comprising an active agent; (9) An emulsion, such as an oil / water / oil emulsion or a water / oil / water emulsion; (10) A micelle; (11) A sphere; (12) A suspension; (13) A dispersion; (14) A vesicle; (15) An aggregate; and (16) A drug carrier comprising any one of (1)-(15) and further comprising one or more layers of material other than the active agent and additional components. In the drug carrier (16), the additional layers may be the outermost layer, the first layer on the core, an intermediate layer interspersed between the layers described in (1)-(15), or any combination thereof. Without limitation, the coating layer may contain components other than those indicated above. For example, the coating components indicated above may be mixed with other molecules or compositions to form the coating layer. This can be used where the specified component cannot form a coating layer on its own. In some embodiments, the particles comprise a core containing an active agent, and additional components form one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) coating layers on the core.

[0175] In some embodiments, the drug carrier may be in the form of a liposome. The liposomes used herein are structures having a lipid membrane that encapsulates a water-containing interior. Liposomes may have one or more lipid membranes. Oligolayer vesicles, macrovesicles, and multilayer vesicles have multiple, typically concentric, membrane layers. Liposomes with several non-concentric membranes (i.e., several smaller vesicles contained within larger vesicles) are referred to as multivesicular vesicles.

[0176] Liposomes may further comprise one or more additional lipids and / or other components (such as sterols, e.g., cholesterol). The additional lipids may be included in the liposome composition for a variety of purposes, such as preventing lipid oxidation, stabilizing the bilayer, reducing aggregation during formation, or attaching ligands to the liposome surface. Any number of additional lipids and / or other components may be present, including amphiphilic lipids, neutral lipids, cationic lipids, anionic lipids, and programmable fusion lipids. Such lipids and / or components may be used alone or in combination. In addition to lipids, liposomes may comprise one or more additives described in this disclosure.

[0177] Liposome compositions can be prepared by a variety of methods known in the art. See, for example: U.S. Patent Nos. 4,235,871; 4,737,323; 4,897,355 and 5,171,678; published international applications WO 96 / 14057 and WO 96 / 37194; Felgner, PL, et al. Proc. Natl. Acad. Sci. American (1987) 8: 7413-7417; Bangham et al. M. Mol. Biol. (1965) 23:238; Olson et al. Biochim. Biophys. Acta (1979) 557:9; Szoka et al., Proc. Natl. Acad. Sci. (1978) 75:4194; Mayhew et al. Biochim. Biophys. Acta (1984) 775:169; Kim et al. Biochim. Biophys. Acta (1983) 728:339; and Fukunaga et al., Endocrinol (1984) 115:757.

[0178] In some embodiments, the drug carrier may be micelles. As used herein, "micelles" refers to a specific type of molecular assembly in which amphiphilic molecules are arranged in a spherical structure such that all hydrophobic portions of the molecules face inwards, leaving the hydrophilic portions in contact with the surrounding water. (The reverse arrangement is not explicitly stated in the original text.)

[0179] In some embodiments, the drug carrier may be an emulsion. As used herein, an "emulsion" is a heterogeneous system in which a liquid is dispersed in droplets within another liquid. Emulsions are often two-phase systems comprising two immiscible liquid phases deeply mixed and dispersed within each other. In the case of emulsion-type ointment bases and creams, either phase in the emulsion may be semi-solid or solid. The active agent may exist as a solution in the aqueous or oil phase, or as the dispersed phase itself.

[0180] In some embodiments, the drug carrier can be formulated as a microemulsion. As used herein, "microemulsion" refers to a system of water, oil, and an amphiphilic compound, which is a single optically isotropic and thermodynamically stable liquid solution. Microemulsions also include thermodynamically stable, isotropic dispersions of two immiscible liquids, stabilized by an interfacial film of surfactant molecules.

[0181] The application and manufacturing methods of emulsion formulations via the skin, mouth, and parenteral routes have been reviewed in the literature, for example, see Idson, in Pharmaceutical Dosage Forms, edited by Lieberman, Rieger, and Banker, 1988, Marcel Dekker, Inc., New York, NY, Vol. 1, p. 199; Rosoff, in Pharmaceutical Dosage Forms, edited by Lieberman, Rieger, and Banker, 1988, Marcel Dekker, Inc., New York, NY, Vol. 1, p. 245; and Block, in Pharmaceutical Dosage Forms, edited by Lieberman, Rieger, and Banker, 1988, Marcel Dekker, Inc., New York, NY, Vol. 1, p. 335, the contents of which are incorporated herein by reference in their entirety.

[0182] Drug carriers can be manufactured using methods and instruments known in the art. For example, drug carriers can be prepared using microprecipitation, encapsulation, depolymerization, a mixture of depolymerization and encapsulation, homogenization, a mixture of depolymerization and thermal homogenization, or any combination thereof. In some embodiments, the process of preparing particles includes the step of selecting particles of the desired size.

[0183] Formulation characteristics applicable to DART, non-DART, and combination APIs

[0184] This disclosure provides compositions or formulations comprising DART. This disclosure also provides compositions or formulations comprising an antibacterial agent as an API, wherein the antibacterial agent is not a DART molecule. In some embodiments, the formulation comprises two or more different APIs, such as two different DARTs, two different non-DART antibacterial agents, or DART molecules and non-DART antibacterial agents. In some embodiments, the DART or antibacterial agent is formulated as a drug delivery vehicle for the API. Without limitation, formulations or compositions can be prepared for administration via any suitable route known in the art, including but not limited to local (including oral and sublingual) 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 limited to, local, injection, infusion, drip, inhalation, or ingestion. "Injection" includes, but is not limited to, intravenous, intramuscular, intraarterial, intrasheath, intravenous, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intralymph node, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, intracerebral, spinal cord, and intracranial injections and infusions. In some embodiments, the formulation may be in the form of an oral dosage form, an injection, an aerosol, or an inhaler.

[0185] In some embodiments, the formulation may contain two or more (e.g., two, three, four, or five or more) different antibacterial agents as an API. For example, the formulation may contain two different anti-acne agents as an API. In some embodiments, the formulation contains 8-chlorobecifloxacin and another anti-acne agent as an API. In one embodiment, the formulation contains becifloxacin and adapalene as APIs.

[0186] The formulations disclosed herein may comprise several types of cosmetically acceptable topical solvents, including but not limited to solutions, colloidal suspensions, dispersions, emulsions (microemulsions, nanoemulsions, multiplexes and non-aqueous emulsions), hydrogels, and vesicles (liposomes, niosomes, novasomes). The composition and formulation methods of suitable cosmetically acceptable topical solvents are well known in the art and described in, for example, U.S. Patent Nos. 6,797,697, 2005 / 0142094 and 2005 / 0008604, and International Patent Application Publications Nos. 2006 / 029818 and 2000 / 062743, all of which are incorporated herein by reference. Various methods for preparing these diverse product forms will be apparent to those skilled in the art.

[0187] In some embodiments, the formulation may be in the form of creams, oils, lotions, serums, gels, sunscreens, nail polishes, ointments, foams, sprays, aerosols, powders, sticks, solutions, suspensions, dispersants, pastes, release agents, and impregnated fabrics (such as “rags” or paper towels). Generally, the composition contains an effective amount of the active agent. As used herein, the term “effective amount” refers to the amount of the formulation containing the active agent necessary to achieve the desired improvement. In some embodiments, the formulation is a topical formulation.

[0188] In some embodiments, the formulation may be in the form of a product selected from the group consisting of: lotions, creams, gels, latexes, oils, serums, powders, sprays, ointments, solutions, suspensions, dispersants, pastes, foams, release agents, films, masks, patches, sticks, rolls, cleansing liquids, facial cleansing sticks, pastes, foams, powders, shaving creams, and fabric-soaked products (e.g., "rags" or paper towels).

[0189] In some embodiments, the formulation is an antibacterial agent. In some embodiments, the composition is an antibacterial composition in the form of a skin care composition. The term "skin care composition" as defined herein refers to a material applied topically to the skin to benefit, improve, or enhance skin conditions, or to treat skin with infections or diseases. Such skin care compositions comprise a matrix, such as a soap base, cosmetic matrix, pharmaceutical matrix, cream matrix, emollient matrix, and combinations thereof, as well as other matrices known in the art.

[0190] Non-limiting, the formulation may contain any desired amount of API. For example, the formulation may contain about 0.01% to about 99% (w / w or w / v) of API. In some embodiments, the formulation may contain about 0.1% to about 75% (w / w or w / v), about 1% to about 50% (w / w or w / v), or about 1.5% to about 40% (w / w or w / v) of API. In some embodiments, the formulation may contain about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, about 7.5%, about 10%, about 12.5%, about 15%, about 17.5%, about 20%, about 22.5%, or about 25% (w / w or w / v) of API.

[0191] In some embodiments, in addition to the API, the formulation may contain one or more zinc compounds. Not wishing to be bound by theory, zinc compounds may help suppress sebum secretion and reduce acne inflammation. Exemplary zinc compounds include, but are not limited to, zinc acetate, zinc methionine, zinc pyrrolidone carboxylate, zinc sulfide, zinc gluconate, zinc pyridinecarboxylate, zinc sulfate, zinc citrate, etc. Without limitation, the formulation may contain any desired amount of zinc compound. For example, the formulation may contain about 0.01% to about 99% (w / w or w / v) of zinc compound. In some embodiments, the formulation may contain about 0.1% to about 75% (w / w or w / v), about 1% to about 50% (w / w or w / v), about 1.5% to about 40% (w / w or w / v), about 2% to about 25% (w / w or w / v), or about 2.5% to about 25% (w / w or w / v) of zinc compound. In some embodiments, the formulation may contain about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5% or 25% (w / w or w / v) of a zinc compound.

[0192] In some embodiments, the formulation may further comprise one or more excipients. Without limitation, excipients may be selected from the group consisting of: emulsifiers, preservatives, surfactants, oils, fats, waxes, stabilizers, rheology modifiers or thickeners (gelling agents), emollients, humectants, conditioning agents, fragrances / perfumes, synergists, preservatives, opacifiers, antioxidants, cooling agents, film-forming agents, scrubs, exfoliants, colorants, pH adjusters, solvents, solvents, penetration enhancers, permeation enhancers, pearlescent agents, and any combination thereof. The amount of excipients in the formulation may range from about 5% to 99.99% (w / w or w / v). In some embodiments, the formulation comprises one or more GRAS (Granitical, Reliable, Assured, Resistant) ingredients.

[0193] Generally, the pH range intended for use in formulations is typically from about pH 2 to about pH 10, from about pH 3 to about pH 9, from about pH 4 to about pH 8, or from about pH 5.0 to about pH 7.5, or from about pH 5 to about pH 6.5. Suitable pH adjusters that can be used include one or more organic or inorganic acids and bases, including sodium hydroxide, potassium hydroxide, ammonium hydroxide, phosphate buffer, citric acid, acetic acid, fumaric acid, hydrochloric acid, malic acid, nitric acid, phosphoric acid, propionic acid, sulfuric acid, tartaric acid, and triethylamine, etc.

[0194] Typically, cosmetically acceptable media for use in skincare compositions include water and other solvents (including, but not limited to, mineral oil and fatty alcohols). The cosmetically acceptable media constitute about 10% to about 99.99% of the composition by weight, preferably about 50% to about 99% by weight, and can achieve a compositional balance in the absence of other additives.

[0195] As used herein, the term "cosmetically acceptable medium" refers to a formulation intended for the treatment of skin, hair, and / or nails, and containing one or more ingredients used by those skilled in the art to formulate skin treatment products. Cosmetically acceptable media may be in any suitable form (i.e., liquid, cream, emulsion, gel, thickening lotion, or powder), typically contains water, and may contain cosmetically acceptable solvents and / or one or more surfactants.

[0196] Formulations may contain one or more conventional functional cosmetic or dermatological additives or adjuvants, provided they do not conflict with the mildness, performance, or aesthetic characteristics desired in the final product. CTFA (The Cosmetic, Toiletry, and Fragrance Association, now known as the Personal Care Products Council) International Cosmetic Ingredient Dictionary and Handbook 11th edition (2006) and McCutcheon's Functional Materials The North American and International Edition, MC Publishing Co. (2007), describes a wide variety of cosmetic and pharmaceutical ingredients commonly used in skincare compositions, suitable for use in the compositions of the present invention. The compositions of the present invention may contain a wide range of additional optional components. The total concentration of the added components is generally less than about 20% of the total weight of the composition, preferably less than about 5% of the total weight of the composition, and most preferably less than about 3% of the total weight of the composition. Such components include, but are not limited to, surfactants, emollients, moisturizers, stabilizers, film-forming substances, fragrances, colorants, chelating agents, preservatives, antioxidants, pH adjusters, antibiotic agents, waterproofing agents, dryness-regulating agents, vitamins, plant extracts, hydroxy acids (e.g., α-hydroxy acids and β-hydroxy acids), and sunscreens.

[0197] The formulation may contain one or more of the following basic cosmetic ingredients, including but not limited to hydrocarbons, esters, fatty alcohols, fatty acids, emulsifiers, humectants, viscosity modifiers, and silicone materials. The formulation may contain a wide range of such basic components. The total concentration of the added ingredients is typically less than 50% of the total weight of the formulation, preferably less than 20%, and most preferably less than 10%. Those skilled in the art will appreciate the various concentrations and combinations of these basic components used to obtain the desired product form.

[0198] Suitable lipids that can be used include one or more hydrocarbons, fatty alcohols, fatty acids, glycerides, or fatty acids with C1-C2. 36 Esters of alkanols. Hydrocarbons may include paraffin or petrolatum. Fatty alcohols may include decanol, dodecyl alcohol, tetradecyl alcohol, hexadecyl alcohol, or octadecyl alcohol. Fatty acids may include C6-C6... 24 Alkyl acids, such as hexanoic acid, caprylic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, and unsaturated fatty acids (such as oleic acid and linoleic acid). Glycerides may include: olive oil; castor oil; sesame oil; caprylic / capric triglycerides; or mono-, di-, and triglycerides of palmitic acid and / or stearic acid. Fatty acid esters may contain C1-C2. 36 Alkyl alcohols, such as beeswax, carnauba wax, hexadecyl palmitate, lanolin, isopropyl myristate, isopropyl stearate, decyl oleate, ethyl oleate, and C6-C... 12 Alkyl esters, etc.

[0199] Suitable hydrocarbons include, but are not limited to, mineral oil, isohexadecane, squalane, hydrogenated polyisobutylene, petrolatum, paraffin, microcrystalline wax, and polyethylene. Suitable oils may include one or more of the following: almond oil, apricot kernel oil, borage oil, canola oil, coconut oil, corn oil, cottonseed oil, fish oil, jojoba soybean oil, lard, linseed oil, boiled macadamia nut oil, mineral oil, olive oil, peanut oil, safflower oil, sesame oil, soybean oil, squalane, sunflower seed oil, tricaprylyl glycerol (1,2,3-tricaprylylglycerol), and wheat germ oil, etc. The preferred amount of oil used ranges from about 5% to about 25% (w / w) of the composition, and more preferably from about 5% to about 20% (w / w) of the composition.

[0200] Suitable esters that can be used include, but are not limited to, isopropyl palmitate, octyl stearate, caprylic / capric triglyceride, vegetable waxes (Canelilla, Caranauba), vegetable oils (natural glycerides), and vegetable oils (jojoba oil).

[0201] Suitable fatty alcohols that can be used include, but are not limited to, myristicin, cetyl alcohol, stearyl alcohol, isostearyl alcohol, and docosanool.

[0202] Suitable emulsifiers that can be used include, but are not limited to, anionic emulsifiers (TEA / K stearate (triethanolamine / potassium stearate), sodium lauryl stearate, sodium cetearyl sulfate, and beeswax / borax), nonionic emulsifiers (glyceryl distearate, PEG (polyethylene glycol)-100 stearate, polysorbate 20, stearyl alcohol polyether 2, and stearyl alcohol polyether 20), and cationic emulsifiers (distearate dimethyl ammonium chloride, behenyl benzyl dimethyl ammonium chloride, and sepiacetium chloride), polymer emulsifiers (acrylate / C10-30 alkyl acrylate crosspolymers, polyacrylamide, polyquaternium-37, propylene glycol, dioctanoate / didecanoate, and PPG-1 tridecyl alcohol polyether-6), and silicone-based materials (alkyl-modified polydimethylsiloxane copolyol), polyglycerol esters, and ethoxylated dicarboxylic acid esters. Other suitable emulsifiers / surfactants may include one or more ionic polysorbate surfactants, Tween... ® 20. Tween ® 40. Tween ® 60. Tween ® 80. Nonylphenol polyethylene glycol ether, alkylphenol-hydroxy polyoxyethylene, poly(oxy-1,2-ethylenedimethyl), α-(4-nonylphenol)-ω-hydroxy, branched (i.e., Tergitol) ® NP-40 surfactant), nonylphenol polyethylene glycol ether mixture (i.e., Tergitol) ® NP-70 (70% AQ) surfactant), phenoxy polyethoxyethanol, and their polymers (e.g., Triton) ® Poloxamer ® Spans ® Tyloxapol ® Different grades of Brij (benzyl sulfide), sodium dodecyl sulfate, etc. The preferred amount of emulsifier / surfactant used is about 0.1% to about 10% (w / w) of the composition.

[0203] Exemplary wetting agents used include, but are not limited to, propylene glycol, sorbitol, butylene glycol, hexanediol, acetamide MEA (acetylethanolamine), honey, sodium PCA (sodium 2-pyrrolidone carboxylate), sorbitol, and triacetin.

[0204] Viscosity modifiers that can be used in the compositions of the present invention include, but are not limited to, xanthan gum, magnesium aluminum silicate, cellulose gum and hydrogenated castor oil.

[0205] Suitable thickeners that can be used include one or more of the following substances: cellulose polymers, carbomer polymers, carbomer derivatives, cellulose derivatives, polyvinyl alcohol, poloxamer, and polysaccharides.

[0206] Suitable emollients may include one or more of the following substances: caprylic / capric triglycerides, castor oil, cetearyl alcohol-20, cetearyl alcohol-30, cetearyl alcohol, cetearyl alcohol polyether 20, cetearyl alcohol, cetearyl alcohol, cetearyl alcohol, cocoa butter, diisopropyl adipate, glycerin, glyceryl monooleate, glyceryl monostearate, glyceryl stearate, isopropyl myristate, isopropyl palmitate, lanolin, lanolin alcohol, hydrogenated lanolin, liquid paraffin, linoleic acid, mineral oil, oleic acid, white petrolatum, polyethylene glycol, polyoxyethylene glycol fatty alcohol ether, polyoxypropylene 15 stearyl ether, propylene glycol stearate, squalane, stearyl alcohol polyether-2 or-100, stearic acid, stearyl alcohol, and urea, etc.

[0207] Suitable preservatives that can be used include one or more of the following substances: phenoxyethanol, parabens (e.g., methylparaben and propylparaben), propylene glycol, sorbates / esters, urea derivatives (e.g., diazolidinyl urea), etc.

[0208] Suitable chelating agents that can be used include one or more of the following substances: disodium EDTA, trisodium edetate, tetrasodium edetate, diethylamine pentaacetate, etc.

[0209] In some embodiments, the formulation comprises one or more alcohols, such as C1-C12 alcohols. 12 Alcohols, diols and triols, glycerol, methanol, ethanol, propanol, octanol, etc.

[0210] In some embodiments, the formulation comprises one or more penetration enhancers. Exemplary penetration enhancers include: anionic surfactants, such as sodium lauryl sulfate and sodium lauryl sulfate; cationic surfactants, such as cetylpyridinium chloride; nonionic surfactants, such as poloxamer, benzyl benzoate, Span, Myrj, and Tween; bile salts; sodium glycodeoxycholate; sodium glycocholate, sodium taurodeoxycholate, sodium taurocholate, and azone. ®); fatty acids, such as oleic acid and caprylic acid; cyclodextrins, such as α-, β-, γ-cyclodextrin, methylated β-cyclodextrin; chelating agents, such as EDTA, sodium citrate, and polyacrylates / esters; polymers, such as deacetylated chitosan, trimethyl chitosan, and cationic amino acids, such as poly-L-arginine and L-lysine. Benzel is a trade name for nonionic polyethylene oxide surfactants available from many suppliers. Span is a trade name for sorbitan surfactants available from many suppliers (e.g., sorbitan trioleate (Span 85) and sorbitan tristearate (Span 65)). Mezzer is a trade name for polyethylene oxide fatty acid surfactants available from many suppliers, such as polyethylene oxide monostearate (Mezzer 49). Tween is the trade name for polyoxyethylene sorbitan or polysorbate surfactant families (such as polyoxyethylene sorbitan trioleate (Tween 85) and polysorbate 80 (Tween 80)) available from many suppliers. Azone is the trade name for L-dodecyl hexahydro-2H-aza-2-one.

[0211] In some embodiments, the formulation comprises one or more penetration enhancers. Exemplary penetration enhancers include, but are not limited to, fatty acids, bile salts, chelating agents, surfactants, and non-surfactants. Exemplary penetration enhancers include dimethyl sulfoxide; isopropyl myristate; decanol, undecylol, or dodecylol; propylene glycol; polyethylene glycol; C9, C10, C11, C12, or C12-15 fatty alcohols; azone; alkylpyrrolidone; diethoxyethylene glycol (Transcutol); lecithin, etc. Surfactants may also be used as penetration enhancers.

[0212] The formulations disclosed herein may further comprise one or more optional components known for use in personal care products, provided that the optional component is physically and chemically compatible with the basic components described herein, or does not unduly impair the stability, appearance, or performance of the product. Individual concentrations of such optional components may range from about 0.001% to about 10% by weight of the composition.

[0213] Non-limiting examples of optional components used in the composition include deposition aids, cationic polymers, nonionic polymers, dispersing granules, conditioning agents (silicones and organic conditioning oils), humectants, suspending agents, additional anti-dandruff actives, viscosity modifiers, dyes, non-volatile solvents or diluents (water-soluble and water-insoluble), pearlescent aids, additional surfactants or nonionic co-surfactants, pediculocides, pH adjusters, fragrances, preservatives, chelating agents, proteins, skin-active agents, sunscreens, UV absorbers, vitamins, antioxidants, preservatives, fillers, surfactants, UVA and / or UVB sunscreens, fragrances, thickeners, humectants, anionic polymers, nonionic polymers, amphoteric polymers, viscosity stabilizers / foam stabilizers, opacifiers / pearlescent agents, blocking agents, stabilizers, humectants, antistatic agents, antifreeze agents, buffers, dyes, and pigments. These adjuvants are well known in the cosmetics industry and have been described in numerous publications, see, for example, [link to relevant documentation]. Harry's Book of Cosmeticology 8th edition, edited by Martin Rieger, Chemical Publishing, New York (2000).

[0214] The compositions disclosed herein may also contain deposition aids. Containing deposition aids effectively enhances the deposition of the composition components. Deposition aids may comprise any material that enhances the deposition of the composition components on hair, scalp, or skin. In some embodiments, the deposition aid is a cationic polymer. The concentration of the deposition aid in the composition should be sufficient to effectively enhance the deposition of the components, and is typically from about 0.05% to about 5% by weight of the composition, preferably from about 0.075% to about 2.5%, more preferably from about 0.1% to about 1.0%.

[0215] The compositions disclosed herein may comprise cationic polymers. The concentration of the cationic polymer in the composition is typically from about 0.05% to about 3% by weight of the composition, preferably from about 0.075% to about 2.0%, more preferably from about 0.1% to about 1.0%. Preferred cationic polymers have a cationic charge density of about 0.9 meq / gm, preferably at least about 1.2 meq / gm, more preferably at least about 1.5 meq / gm. Such suitable cationic polymers typically have an average molecular weight between about 10,000 and 10 million, preferably between about 50,000 and about 5 million, more preferably between about 100,000 and about 3 million.

[0216] Suitable cationic polymers for use in compositions comprise a cationic nitrogen-containing moiety containing, for example, a quaternary ammonium or a cationic protonated amino group. The cationic protonated amine can be a primary, secondary, or tertiary amine (preferably secondary or tertiary), depending on the specific species and the pH selected for the composition. Any anionic counterion can be used with the corresponding cationic polymer, provided that the polymer remains water-soluble in the composition or in the condensed phase of the composition, and provided that the counterion is physically and chemically compatible with the basic components of the composition, or does not unduly impair the performance, stability, and aesthetics of the product. Non-limiting examples of such counterions include halide ions (e.g., chloride, fluoride, bromide, iodide), sulfate, and methyl sulfate. Non-limiting examples of cationic polymers are described in the CTFA Dictionary of Cosmetic Ingredients, 3rd Edition, edited by Estrin, Crosley, and Haynes (The Cosmetic, Toiletry, and Fragrance Association, Inc., Washington, DC, (1982)).

[0217] Non-limiting examples of suitable cationic polymers include copolymers of vinyl monomers having cationic protonated amine or quaternary ammonium functional groups with water-soluble spacer monomers (e.g., acrylamide, methacrylamide, alkyl and dialkyl acrylamide, alkyl and dialkyl methacrylamide, alkyl acrylate, alkyl methacrylate, vinyl caprolactone, or vinylpyrrolidone).

[0218] Suitable cationic protonated amino and quaternary ammonium monomers for inclusion in the cationic polymers of the compositions of the present invention include: vinyl compounds substituted with dialkylaminoalkyl acrylates, dialkylaminoalkyl methacrylates, monoalkylaminoalkyl acrylates, monoalkylaminoalkyl methacrylates, or trialkylmethacryloyloxyalkylammonium salts; and vinyl quaternary ammonium monomers having a cyclic cationic nitrogen-containing ring (e.g., pyridinium, imidazolium, and quaternized pyrrolidone), such as alkylvinylimidazolium, alkylvinylpyridinium, or alkylvinylpyrrolidone salts.

[0219] Other suitable cationic polymers for use in the composition include copolymers of 1-vinyl-2-pyrrolidone and 1-vinyl-3-methylimidazolium salts (e.g., chloride salts) (from Cosmetic, Toiletry, and Fragrance Association (“CTFA”) refers to the following in the industry: polyquaternium-16; copolymers of 1-vinyl-2-pyrrolidone and dimethylaminoethyl methacrylate (referred to in the industry by CTFA as polyquaternium-11); cationic diallyl quaternary salts containing polymers, including, for example, homopolymers of dimethyl diallyl chloride, copolymers of acrylamide and dimethyl diallyl chloride (referred to in the industry by CTFA as polyquaternium-6 and polyquaternium-7, respectively); amphoteric copolymers of acrylic acid, including copolymers of acrylic acid and dimethyl diallyl chloride (referred to in the industry by CTFA as polyquaternium-22), terpolymers of acrylic acid with dimethyl diallyl chloride and acrylamide (referred to in the industry by CTFA as polyquaternium-39), and terpolymers of acrylic acid with methacrylamide propyltrimethylammonium chloride and methyl acrylate (referred to in the industry by CTFA as polyquaternium-47).

[0220] 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 hydroxyethyl cellulose reacted with trimethylammonium-substituted epoxides, known in the industry (CTFA) as polyquaternium-10, and are available from Amerchol Corp. (Edison, NJ, USA) in their polymer series LR, JR, and KG. Other suitable types of cationic cellulose include polymeric quaternium salts of hydroxyethyl cellulose reacted with lauryl dimethylammonium-substituted epoxides, known in the industry (CTFA) as polyquaternium-24. These materials are available from Amerchol Corp. under the trade name Polymer LM-200.

[0221] Other suitable cationic polymers include cationic guar gum and its derivatives, such as guar hydroxypropyltrimethylammonium chloride, specific examples of which include the Jaguar series commercially available from Rhone-Poulenc Incorporated and the N-Hance series commercially available from Aqualon Division of Hercules, Inc. Other suitable cationic polymers include quaternary nitrogen-containing cellulose ethers, some examples of which are described in U.S. Patent No. 3,962,418. Other suitable cationic polymers include copolymers of etherified cellulose, guar, and starch, some examples of which are described in U.S. Patent No. 3,958,581. When used, the cationic polymers herein are soluble in the composition or in a complex condensed phase soluble in the composition, which is formed by the cationic polymers described above with anionic, amphoteric, and / or zwitterionic detergency surfactant components. The complex condensate of the cationic polymers may also be formed by other charged substances in the composition.

[0222] Polyalkylene glycols with a molecular weight greater than approximately 1000 are useful in this paper. One useful polyethylene glycol polymer in this paper is PEG-2M (also known as Polyox WSR). ® N-10, purchased from Union Carbide as PEG-2,000; PEG-5M (also known as Polyox WSR) ® N-35 and Polyox WSR ® N-80, purchased from UnionCarbide as PEG-5000 and polyethylene glycol 300,000; PEG-7M (also known as Polyox WSR) ® N-750, purchased from Union Carbide; PEG-9M (also known as Polyox WSR) ® N-3333, purchased from Union Carbide); and PEG-14 M (also known as Polyox WSR). ® N-3000, purchased from UnionCarbide.

[0223] The composition may also contain dispersed particles. The composition may contain at least 0.025% (by weight) of dispersed particles, more preferably at least 0.05%, even more preferably at least 0.1%, even more preferably at least 0.25%, and even more preferably at least 0.5% (by weight) of dispersed particles. In some embodiments, it is preferred to incorporate no more than about 20% (by weight) of dispersed particles, more preferably no more than about 10%, even more preferably no more than 5%, even more preferably no more than 3%, and even more preferably no more than 2% (by weight) of dispersed particles.

[0224] Conditioning agents include any material used to provide specific conditioning benefits to the skin. Conditioning agents useful in the compositions of the present invention typically comprise water-insoluble, water-dispersible, non-volatile liquids that form emulsified liquid particles or are solubilized in anionic detergency surfactant components (described above) via surfactant micelles. Suitable conditioning agents used in the compositions are: conditioning agents typically characterized as silicones (e.g., silicone oils, cationic silicones, silicone gels, high-refractive-index silicones, and silicone resins), organic conditioning oils (e.g., hydrocarbon oils, polyolefins, and fatty esters), or combinations thereof; or conditioning agents that form liquid, dispersed particles in the aqueous surfactant matrix described herein.

[0225] The conditioning agent in the composition can be an insoluble silicone conditioning agent. The silicone conditioning agent particles may contain volatile silicone, non-volatile silicone, or a combination thereof. Non-volatile silicone conditioning agents are preferred. If volatile silicone is present, it is typically used as a solvent or carrier in the commercially available form of non-volatile silicone material components (e.g., silicone gels and resins). The silicone conditioning agent particles may contain silicone fluid conditioning agents, or other components (e.g., silicone resins), to improve the deposition efficiency of silicone fluids or enhance hair shine.

[0226] The concentration of the silicone conditioner is typically from about 0.01% to about 10% of the composition weight, preferably from about 0.1% to about 8%, more preferably from about 0.1% to about 5%, and even more preferably from about 0.2% to about 3%. Non-limiting examples of suitable silicone conditioners and optional suspending agents for silicone are described in U.S. Reissue Patent No. 34,584, U.S. Patent No. 5,104,646, and U.S. Patent No. 5,106,609. The viscosity of the silicone conditioner used in the compositions of the present invention is preferably (measured at 25°C) from about 20 centistokes (“csk”) to about 2,000,000 csk, more preferably from about 1,000 csk to about 1,800,000 csk, even more preferably from about 50,000 csk to about 1,500,000 csk, and even more preferably from about 100,000 csk to about 1,500,000 csk.

[0227] The volume average particle size of the dispersed silicone conditioner particles typically ranges from about 0.01 μm to about 50 μm. For small particles applied to hair, the volume average particle size is typically from about 0.01 μm to about 41 μm, preferably from about 0.01 μm to about 2 μm, and more preferably from about 0.01 μm to about 0.51 μm. For larger particles applied to hair, the volume average particle size is typically from about 5 μm to about 125 μm, preferably from about 10 μm to about 90 μm, more preferably from about 15 μm to about 70 μm, and more preferably from about 20 μm to about 50 μm.

[0228] exist Encyclopedia of Polymer Science and EngineeringBackground material on silicones can be found in John Wiley & Sons, Inc. (1989), Volume 15, 2d ed., pp. 204-308, including a discussion of silicone fluids, adhesives and resins and the preparation of silicones.

[0229] Silicone fluids include silicone oils, which are flowable silicone materials with a viscosity (measured at 25°C) of less than 1,000,000 csk, preferably about 5 csk to about 1,000,000 csk, more preferably about 100 csk to about 600,000 csk. Suitable silicone oils used in the compositions of the present invention include polyalkylsiloxanes, polyarylsiloxanes, polyalkylarylsiloxanes, polyether siloxane copolymers, and mixtures thereof. Other insoluble, non-volatile silicone fluids with hair conditioning properties may also be used.

[0230] Other silicone fluids suitable for the composition are insoluble silicone sealants. These sealants are polyorganosiloxane materials with a viscosity greater than or equal to 1,000,000 csk (measured at 25°C). Silicone sealants are described in the following literature: US Patent No. 4,152,416; Noll and Walter, Chemistry and Technology of Silicones New York: Academic Press (1968); and General Electric Silicone Rubber Product Data Sheets SE 30, SE 33, SE 54 and SE 76. Specific, non-limiting examples of silicone sealants used in this invention include polydimethylsiloxane, (polydimethylsiloxane)(methylvinylsiloxane) copolymers, (polydimethylsiloxane)(diphenylsiloxane)(methylvinylsiloxane) copolymers, and mixtures thereof.

[0231] Other non-volatile, insoluble silicone fluid conditioners suitable for use in the compositions of the present invention are reagents referred to as "high refractive index silicones" having a refractive index of at least about 1.46, preferably at least about 1.48, more preferably at least about 1.52, and even more preferably at least about 1.55. Polysiloxane fluids generally have a refractive index of less than about 1.70, typically less than about 1.60. In this context, polysiloxane "fluids" include oils and adhesives.

[0232] Silicone fluids suitable for use in the compositions of the present invention are disclosed in U.S. Patent Nos. 2,826,551, 3,964,500, 4,364,837, and 849,433, UK Patent No. Silicon Compounds , Petrarch Systems, Inc. (1984).

[0233] Silicone resins may be included in the silicone conditioning agents of the compositions of the present invention. These resins are highly cross-linked polymeric siloxane systems. Cross-linking is introduced during the manufacture of silicone resins by incorporating monofunctional or difunctional silanes, or both, into trifunctional and tetrafunctional silanes.

[0234] Silicone materials, particularly silicone resins, can be conveniently identified using a shorthand nomenclature system (referred to as the "MDTQ" nomenclature) known to those skilled in the art. In this system, silicones are described based on the presence of various siloxane monomer units that constitute the silicone. Simply put, the symbol M represents the monofunctional unit (CH3)3SiO. 05 D represents the difunctional unit (CH3)2SiO; T represents the trifunctional unit (CH3)SiO. 15 ; and Q represents the quaternary or tetrafunctional unit SiO2. Subscript symbols of the unit symbols (e.g., M', D', T and Q') represent substituents other than methyl groups and must be specifically defined each time they appear.

[0235] Preferred silicone resins used in the compositions of the present invention include, but are not limited to, MQ, MT, MTQ, MDT, and MDTQ resins. Methyl groups are preferred silicone substituents. Particularly preferred silicone resins are MQ resins, wherein the M:Q ratio is from about 0.5:1.0 to about 1.5:1.0, and the average molecular weight of the silicone resin is from about 1000 to about 10,000.

[0236] The composition of the present invention may further comprise about 0.05% to about 3% by weight of a conditioning component, preferably about 0.08% to about 1.5%, more preferably about 0.1% to about 1%, of at least one organic conditioning oil, alone or in combination with other conditioning agents (e.g., silicone (as described above)) as a conditioning agent.

[0237] Suitable organic conditioning oils for use as conditioning agents in the compositions of the present invention include, but are not limited to, hydrocarbon oils having at least about 10 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 are preferably about C to about C19. Branched-chain hydrocarbon oils (including hydrocarbon polymers) generally contain more than 19 carbon atoms.

[0238] Specific, non-limiting examples of these hydrocarbon oils include paraffin oils, mineral oils, saturated and unsaturated dodecane, saturated and unsaturated tridecane, saturated and unsaturated tetradecane, saturated and unsaturated pentadecane, saturated and unsaturated hexadecane, polybutene, polydecene, and mixtures thereof. Branched isomers of these compounds, as well as hydrocarbons with higher chain lengths, can also be used, examples of which include highly branched, saturated or unsaturated alkanes, such as permethyl-substituted isomers, such as permethyl-substituted isomers of hexadecane and eicosane, such as 2,2,4,4,6,6,8,8-dimethyl-10-methylundecane and 2,2,4,4,6,6-dimethyl-8-methylnonane, available from Permethyl. Hydrocarbon polymers (e.g., polybutene and polydecene) are preferred. A preferred hydrocarbon polymer is polybutene, such as a copolymer of isobutene and butene. Commercially available materials of this type are L-14 polybutene from Amoco Chemical Corporation.

[0239] The organic conditioning oil used in the compositions of the present invention may also include liquid polyolefins, more preferably liquid polyalphaolefins, and more preferably hydrogenated liquid polyalphaolefins. The polyolefins used herein are prepared by polymerization of C4 to C14 olefin monomers, preferably about C6 to about C12 olefin monomers.

[0240] Non-limiting examples of olefin monomers used in the preparation of the polyolefin liquids described herein include ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, 1-decene, 1-dodecene, 1-tetradecene, branched isomers (e.g., 4-methyl-1-pentene), and mixtures thereof. Also suitable for preparing polyolefin liquids are olefins comprising refinery feedstocks or effluents. Preferred hydrogenated α-olefin monomers include, but are not limited to, 1-hexene to 1-hexadecene, 1-octene to 1-tetradecene, and mixtures thereof.

[0241] Other suitable organic conditioning oils used as conditioning agents in the compositions of the present invention include, but are not limited to, aliphatic esters having at least 10 carbon atoms. These aliphatic esters include esters having hydrocarbon chains derived from fatty acids or alcohols (e.g., monoesters, polyol esters, and dicarboxylic acid esters and tricarboxylic acid esters). Here, the hydrocarbon radical of the aliphatic ester may include or have covalent bonds (e.g., ethoxy or ether bonds, etc.) bonded to other compatible functional groups, such as amide and alkoxy moieties.

[0242] Preferred examples of aliphatic esters include, but are not limited to: isopropyl isostearate, hexyl laurate, isohexyl laurate, isohexyl palmitate, isopropyl palmitate, decyl oleate, isodecyl oleate, hexadecyl stearate, decyl stearate, dihexyl decyl adipate, lauryl lactate, myristyl lactate, cetyl lactate, oleyl stearate, oleyl oleate, myristate oleate, lauryl acetate, cetyl propionate, and oleyl adipate.

[0243] Other aliphatic esters used in the compositions of the present invention are monocarboxylic acid esters of the general formula R'COOR, wherein R' and R are alkyl or alkenyl groups, and the sum of the number of carbon atoms in R' and R is at least 10, preferably at least 22.

[0244] Other aliphatic esters suitable for use in the compositions of the present invention are dialkyl, trialkyl, and alkenyl esters of carboxylic acids, such as C4 to C8 esters of dicarboxylic acids (e.g., C1 to C22 esters of succinic acid, glutaric acid, and adipic acid, preferably C1 to C6 esters). Specific, non-limiting examples of dialkyl, trialkyl, and alkenyl esters of carboxylic acids include isocetyl stearoyl stearate, diisopropyl adipate, and tristearyl citrate. In some embodiments, the composition comprises an ester of at least one of lauric acid and succinic acid having additional anti-acne and / or anti-inflammatory properties.

[0245] Other aliphatic esters suitable for use in the compositions of the present invention are known polyol esters. Such polyol esters include alkylene glycol esters, such as ethylene glycol esters of mono- and di-fatty acids, diethylene glycol esters of mono- and di-fatty acids, polyethylene glycol esters of mono- and di-fatty acids, propylene glycol esters of mono- and di-fatty acids, polypropylene glycol monooleate, polypropylene glycol monostearate 2000, ethoxylated propylene glycol monostearate, glyceryl esters of mono- and di-fatty acids, polyglyceryl polyesters of fatty acids, ethoxylated glyceryl monostearate, 1,3-butanediol monostearate, 1,3-butene glycol distearate, polyoxyethylene polyoxyethylene esters of hydroxyl fatty acids, sorbitan esters of fatty acids, and sorbitan esters of polyoxyethylene polyoxyethylene esters of fatty acids.

[0246] Other aliphatic esters suitable for use in the compositions of the present invention are glycerides, including but not limited to monoglycerides, diglycerides, and triglycerides, preferably diglycerides and triglycerides, more preferably triglycerides. When used in the compositions described herein, the glycerides are preferably monoesters, diesters, and triesters of glycerol and long-chain carboxylic acids (e.g., C10-C22 carboxylic acids). Various types of these materials are available 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 include, but are not limited to, triolein and tristearin dilaurate.

[0247] Other aliphatic esters suitable for use in the compositions of the present invention are water-insoluble synthetic aliphatic esters.

[0248] Specific, non-limiting examples of suitable synthetic aliphatic esters used in the compositions of the present invention include P-43 (a C8-C10 triester of trimethylolpropane), MCP-684 (a tetraester of 3,3-dihydroxyethyl-1,5-pentanediol), and MCP 121 (a C8-C10 diester of adipic acid), all of which are available from Mobil Chemical Company.

[0249] Also suitable for use in the compositions of the present invention are the conditioning agents described by Procter & Gamble in U.S. Patent Nos. 5,674,478 and 5,750,122. Also suitable for use herein are the conditioning agents described in the following documents: U.S. Patent Nos. 4,529,586 (Clairol), 4,507,280 (Clairol), 4,663,158 (Clairol), 4,197,865 (L'Oreal), 4,217,914 (L'Oreal), 4,381,919 (L'Oreal), and 4,422,853 (L'Oreal).

[0250] The composition may contain a wetting agent. The wetting agent may be selected from the group consisting of polyols, water-soluble alkoxylated nonionic polymers, and mixtures thereof. When used herein, the wetting agent is preferably used at a level of about 0.1% to about 20%, more preferably about 0.5% to about 5% by weight of the composition.

[0251] The polyols useful in this article include glycerol, sorbitol, propylene glycol, butylene glycol, hexanediol, ethoxylated glucose, 1,2-hexanediol, glycerol, dipropylene glycol, erythritol, trehalose, diglycerol, xylitol, maltitol, maltose, glucose, fructose, sodium chondroitin sulfate, sodium hyaluronate, sodium adenosine phosphate, sodium lactate, pyrrolidone carbonate, glucosamine, cyclodextrin, and mixtures thereof.

[0252] Useful water-soluble alkoxylated nonionic polymers in this article include polyethylene glycol and polypropylene glycol with molecular weights up to about 1000, such as polymers with CTFA names PEG-200, PEG-400, PEG-600, PEG-1000, and mixtures thereof.

[0253] The compositions of the present invention may further comprise a suspending agent at an effective concentration for suspending the water-insoluble material in dispersed form in the composition or for improving the viscosity of the composition. This concentration ranges from about 0.1% to about 10%, preferably from about 0.3% to about 5.0% by weight of the composition.

[0254] Suitable suspending agents include crystalline suspending agents, which can be classified as acyl derivatives, long-chain amine oxides, or combinations thereof. These suspending agents are described in U.S. Patent No. 4,741,855.

[0255] The composition may also contain vitamins and amino acids, such as: water-soluble vitamins, such as vitamins B1, B2, B6, B12, C, pantothenic acid, ubiquitin ethyl ether, panthenol, biotin, and their derivatives; water-soluble amino acids, such as asparagine, alanine, tryptophan, glutamic acid, and their salts; water-insoluble vitamins, such as vitamins A, D, E, and their derivatives; and water-insoluble amino acids, such as tyrosine, tryptophan, and their salts.

[0256] The formulations disclosed herein may also contain pigment materials, such as nitroso compounds, monoazo compounds, diazo compounds, carotenoids, triphenylmethane, triarylmethane, xanthones, quinoline, oxazine, azazine, anthraquinones, indigo derivatives, thioindigo derivatives, quinacridones, phthalocyanines, plant pigments, and natural pigments comprising water-soluble dye components. The compositions of the present invention may also contain chelating agents.

[0257] In one embodiment, the formulation is a moisturizing cream / gel base. For example, the formulation contains at least one moisturizer. Typically, the formulation may contain from about 0.01% (by weight) to about 50% (by weight) of a moisturizer to provide moisturizing benefits upon application. It should be noted that dryness is one of the primary concerns known for topical anti-acne products. Exemplary moisturizers include, but are not limited to: N-acetylethanolamine, aloe vera gel, arginine PCA, chitosan PCA, copper PCA, corn glyceride, dimethylimidazolidine, fructose, glucosamine, glucose, glucosamine glutamate, glucuronic acid, glutamic acid, glyceryl polyether-7, glyceryl polyether-12, glyceryl polyether-20, glyceryl polyether-26, glycerin, honey, hydrogenated honey, hydrogenated starch hydrolysate, hydrolyzed corn starch, lactamide MEA, lactic acid, lactose lysine PCA, mannitol, methylglutamic acid polyether-10, methylglutamic acid polyether-20, P CA, PEG-2 lactamide, PEG-10 propylene glycol, polyamino acids, polysaccharides, polyamino sugar condensates, potassium PCA, propylene glycol, propylene glycol citrate, sugar hydrolysate, isomers, sodium aspartate, sodium lactate, sodium PCA, sorbitol, TEA-lactic acid ester, TEA-PCA, urea, xylitol, panthenol, petrolatum, mineral oil, lanolin, lanolin alcohol, tocopherol, tocopheryl ester, alkyl polydimethylsiloxane, vegetable oil, hydrogenated vegetable oil, fatty acid esters, beeswax, hydrolyzed keratin, hydroxyethyl urea, carboxylic acid amide, mucopolysaccharides, and quaternary nitrogen humectants. Examples of quaternary nitrogen humectants include, but are not limited to, hydroxypropyl dihydroxyethyl dimethyl ammonium chloride (available as COLA™ Moist 200 from Colonial Chemicals, Inc.), humectants described in U.S. Patent No. 6,869,977 (the contents of which are incorporated herein by reference), choline salts described in U.S. Patent Nos. 6,475,965 and 6,265,364 (the contents of which are incorporated herein by reference), carnitine, and combinations thereof. Humectants may be present in the formulation in any desired amount to impart a desired level of moisturizing effect. In some embodiments, the humectant may be present in an amount of 0 to about 5%. In another embodiment, the quaternary nitrogen humectant is present in an amount of about 0.1% to about 1% by weight. In yet another embodiment, the humectant is present in an amount of about 1% by weight.

[0258] In some embodiments, the formulation comprises at least one of glycolic acid, lactic acid, sulfur, salicylic acid, and resorcinol.

[0259] Some exemplary formulations are described in Tables 2-5.

[0260] Table 2: Some exemplary cream formulations

[0261]

[0262] Table 3: Some exemplary latex formulations

[0263]

[0264] Table 4: Some exemplary gel formulations

[0265]

[0266] Table 5: Some exemplary lotion formulations

[0267]

[0268] Without being bound by theory, the formulation disclosed herein exhibits at least a 1.2-fold increase in the area under the curve (AUC) on the skin concentration-time plot compared to formulations known in the art. Furthermore, this formulation kills at least 20% more Propionibacterium acnes compared to direct antibiotic application.

[0269] The formulations disclosed herein represent advancements in formulation technology (size optimization, surface modification, and formulation innovation) that improve specificity and potency by enhancing penetration and delivery to target sites (sebaceous glands); improve retention for sustained action; or facilitate entry into biofilm-encapsulated bacteria.

[0270] This invention also provides the use of the DART and formulations disclosed herein in treating or preventing at least one bacterial infection condition in a subject. The method generally involves administering the disclosed DART or formulation to a subject in need. In some embodiments, the method is used to treat acne in a subject.

[0271] The term "acne" includes an inflammatory disease of the pilosebaceous follicles and / or sebaceous glands, and is typically characterized by papules, pustules, cysts, nodules, comedones, other blemishes, or skin lesions. As used herein, the term "acne" includes all known types of acne. Some types of acne that can be treated with the compositions of the present invention include: acne vulgaris, comedonal acne, papular acne, premenstrual acne, prepubertal acne, and toxic acne. (venenata), cosmetic acne, hair cream acne, detergent acne, exfoliative acne, Gram-negative acne, pseudofolliculitis of the beard, folliculitis, perioral dermatitis, hidradenitis suppurativa, cystic acne, atrophic acne, bromoacne, chloracne, conglobate acne, detergent acne, epidemic acne, summer acne, fulminant acne, halogenated acne, indurated acne, iodine acne, scarring acne, mechanical acne, papular acne, hair cream acne, premenstrual acne, pustular acne, scurvy acne, adenoid acne, urticarial acne, acne vulgaris, toxic acne, propionic acid acne, exfoliative acne, Gram-negative acne, steroid acne, nodular cystic acne, and rosacea.

[0272] To avoid being bound by theory, micronization of besifloxacin can affect its biological activity. For example, micronization can enhance the biological activity of besifloxacin or promote its retention at the desired site. Furthermore, micronization can affect the stability and amount of besifloxacin in formulations. Additionally, micronization can allow for the optimization of the properties of formulations containing micronized besifloxacin.

[0273] Implementations of the various aspects disclosed herein may also be described in one or more numbered paragraphs:

[0274] 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 delivery system comprising the anti-acne agent and at least one additional compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0275] 2. The formulation as described in paragraph 1, wherein the drug carrier is coated or uncoated.

[0276] 3. The formulation as described in paragraph 1 or 2, wherein the size of the drug delivery body is from about 5 nm to about 20 μm.

[0277] 4. The formulation as described in paragraph 1 or 3, wherein the size of the drug carrier is from about 5 nm to about 5 μm.

[0278] 5. The formulation as described in any of the preceding paragraphs, the formulation further comprising a surface modifier located on the surface of the drug carrier.

[0279] 6. The formulation as described in any one of paragraphs 1-5, wherein the surface modifier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0280] 7. The formulation as described in any one of paragraphs 1-6, wherein the carrier or excipient is selected from the group consisting of: emulsifiers, preservatives, surfactants, oils, fats, waxes, stabilizers, rheology modifiers or thickeners (gelling agents), emollients, humectants, conditioning agents, fragrances / perfumes, synergists, preservatives, opacifiers, antioxidants, cooling agents, film-forming agents, exfoliants, scabies, colorants, pH adjusters, solvents, solvents, penetration enhancers, pearlescent agents, and any combination thereof.

[0281] 8. The formulation as described in any of paragraphs 1-4, wherein the surface of the drug carrier is substantially free of surface modifiers.

[0282] 9. The formulation as described in any of paragraphs 1-8, wherein the formulation comprises about 0.1% to about 50% (w / w or w / v) of the carrier or excipient.

[0283] 10. The formulation as described in any of paragraphs 1-9, wherein the formulation is formulated for topical, oral or parenteral administration.

[0284] 11. The formulation as described in any of paragraphs 1-10, wherein the formulation is an oral dosage form, an injection, an aerosol or inhaler, a lotion, a cream, a gel, a latex, an oil, a serum, a powder, a spray, an ointment, a solution, a suspension, a dispersant, a paste, a foam, a release agent, a film, a mask, a patch, a stick, a roll-on, a fabric soaked in fabric (e.g., a "rag" or a paper towel), or any combination thereof.

[0285] 12. The formulation as described in any of paragraphs 1-11, wherein the formulation further comprises a second anti-acne agent.

[0286] 13. The formulation as described in any one of paragraphs 1-12, wherein the second anti-acne agent is selected from the group consisting of: 8-chlorofluoroquinolones, acitretin, adapalene, retinoic acid, α-hydroxy acids or β-hydroxy acids, antibiotics, antimicrobial peptides, antimicrobial agents, azelaic acid, benzoyl peroxide, bexarotine, bile salts, biofilm inhibitors, clindamycin, erythromycin, etretinate, glycolic acid, isotretinoin, keratolytic agents, lactic acid, lipoic acid, N-acetylcysteine, natural anti-acne agents, oxymethopyrone, phenoxyethanol, phenoxypropanol, pyruvate, resorcinol, retinoic acid, retinoids, salicylic acid, sebostats, sodium sulfacetamide, spironolactone, sulfur, sulfur-containing D- or L-amino acids, tazarotene, tea tree oil, retinoic acid, triclosan, urea, and any combination thereof.

[0287] 14. The formulation as described in any of paragraphs 1-13, wherein the formulation comprises 8-chlorofluoroquinolone alone, or a combination of 8-chlorofluoroquinolone and another anti-acne agent.

[0288] 15. The formulation as described in any of paragraphs 1-14, wherein the formulation comprises besifloxacin and adapalene.

[0289] 16. The formulation as described in any of paragraphs 1-14, wherein the formulation comprises 8-chlorofluoroquinolone and an anti-inflammatory agent.

[0290] 17. The formulation as described in any of paragraphs 1-14, wherein the formulation comprises 8-chlorofluoroquinolone and retinoic acid or retinoids.

[0291] 18. The formulation as described in any of paragraphs 1-17, 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 compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0292] 19. The formulation as described in any of paragraphs 1-18, wherein the size of the second anti-acne agent carrier is from about 5 nm to about 50 μm.

[0293] 20. The formulation as described in any of paragraphs 1-19, wherein the size of the second anti-acne agent carrier is from about 100 nm to about 25 μm.

[0294] 21. The formulation as described in any of paragraphs 1-20, wherein the second anti-acne agent carrier comprises a surface modifier located on its surface.

[0295] 22. The formulation as described in any of paragraphs 1-21, wherein the surface modifier of the second anti-acne agent carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0296] 23. The formulation as described in any of paragraphs 1-20, wherein the surface of the second anti-acne agent carrier is substantially free of surface modifiers.

[0297] 24. The formulation as described in any of paragraphs 1-23, wherein the formulation further comprises an additional active agent.

[0298] 25. The formulation as described in any of paragraphs 1-24, wherein the additional active agent is an anti-inflammatory agent, a penetration enhancer, an antioxidant, an anti-aging agent, an anti-wrinkle agent, a skin brightener or bleaching agent, an ultraviolet (UV) absorber or scattering agent, a skin depigmenting agent, a skin regenerating agent, a scar healing agent, or any combination thereof.

[0299] 26. The formulation as described in any of paragraphs 1-25, wherein the additional active agent is in the form of a drug carrier comprising a compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0300] 27. The formulation as described in any of paragraphs 1-26, wherein the size of the additional active agent carrier is from about 5 nm to about 100 μm.

[0301] 28. The formulation as described in any of paragraphs 1-27, wherein the size of the additional active agent carrier is from about 100 nm to about 25 μm.

[0302] 29. The formulation as described in any of paragraphs 1-28, wherein the additional active agent carrier comprises a surface modifier located on its surface.

[0303] 30. The formulation as described in any of paragraphs 1-29, wherein the surface modifier of the additional active agent carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0304] 31. The formulation as described in any of paragraphs 1-30, wherein the surface of the additional active agent carrier is substantially free of surface modifiers.

[0305] 32. The formulation as described in any of paragraphs 1-31, wherein the formulation further comprises a zinc compound.

[0306] 33. A formulation comprising an antibacterial agent and at least one carrier or excipient, wherein the antibacterial agent is in the form of a drug delivery system comprising the antibacterial agent and at least one additional compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0307] 34. The formulation as described in paragraph 33, wherein the size of the drug delivery body is from about 5 nm to about 100 μm.

[0308] 35. The formulation as described in paragraph 33 or 34, wherein the size of the drug carrier is from about 100 nm to about 25 μm.

[0309] 36. The formulation as described in any of paragraphs 33-35, the formulation further comprising a surface modifier located on the surface of the drug carrier.

[0310] 37. The formulation as described in any of paragraphs 33-36, wherein the surface modifier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0311] 38. The formulation as described in any of paragraphs 33-37, wherein the surface of the drug carrier is substantially free of surface modifiers.

[0312] 39. The formulation as described in any of paragraphs 33-38, wherein the carrier or excipient is selected from the group consisting of: emulsifiers, preservatives, surfactants, oils, fats, waxes, stabilizers, rheology modifiers or thickeners (gelling agents), emollients, humectants, conditioning agents, fragrances / flavors, synergists, preservatives, opacifiers, antioxidants, cooling agents, film-forming agents, exfoliants, scabies, colorants, pH adjusters, solvents, solvents, penetration enhancers, pearlescent agents, and any combination thereof.

[0313] 40. The formulation as described in any of paragraphs 33-39, wherein the formulation comprises about 5% to about 99% (w / w or w / v) of the carrier or excipient.

[0314] 41. The formulation as described in any of paragraphs 33-40, wherein the formulation is formulated for topical administration, oral administration or parenteral administration.

[0315] 42. The formulation as described in any of paragraphs 33-41, wherein the formulation is an oral dosage form, an injection, an aerosol or inhaler, a lotion, a cream, a gel, a latex, an oil, a serum, a powder, a spray, an ointment, a solution, a suspension, a dispersant, a paste, a foam, a release agent, a film, a mask, a patch, a stick, a roll-on, a fabric soaked in fabric (e.g., a "rag" or a paper towel), or any combination thereof.

[0316] 43. The formulation as described in any of paragraphs 33-42, wherein the formulation further comprises a second antibacterial agent.

[0317] 44. The formulation as described in any of paragraphs 33-43, wherein the second antibacterial agent is in the form of a drug carrier.

[0318] 45. The formulation as described in any of paragraphs 33-44, wherein the second antibacterial agent carrier further comprises a compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0319] 46. ​​The formulation as described in any of paragraphs 33-45, wherein the size of the second antibacterial agent carrier is from about 5 nm to about 100 μm.

[0320] 47. The formulation as described in any of paragraphs 33-46, wherein the size of the second antibacterial agent carrier is from about 100 nm to about 25 μm.

[0321] 48. The formulation as described in any of paragraphs 33-47, wherein the second antibacterial agent carrier comprises a surface modifier located on its surface.

[0322] 49. The formulation as described in any of paragraphs 33-48, wherein the surface modifier of the second antibacterial agent carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0323] 50. The formulation as described in any of paragraphs 33-47, wherein the surface of the second antibacterial agent carrier is substantially free of surface modifiers.

[0324] 51. The formulation as described in any of paragraphs 33-50, wherein the formulation further comprises an additional active agent.

[0325] 52. The formulation as described in any of paragraphs 33-51, wherein the additional active agent is an anti-inflammatory agent, a penetration enhancer, a permeation enhancer, an antioxidant, an anti-aging agent, an anti-wrinkle agent, a skin whitening agent or bleaching agent, an ultraviolet (UV) absorber or scattering agent, a skin depigmenting agent, a skin regenerating agent, a scar healing agent, or any combination thereof.

[0326] 53. The formulation as described in any of paragraphs 33-52, wherein the additional active agent is in the form of a drug carrier.

[0327] 54. The formulation as described in any of paragraphs 33-53, wherein the additional active agent carrier further comprises a compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0328] 55. The formulation as described in any of paragraphs 33-54, wherein the size of the additional active agent carrier is from about 5 nm to about 100 μm.

[0329] 56. The formulation as described in any of paragraphs 33-55, wherein the size of the additional active agent carrier is from about 100 nm to about 25 μm.

[0330] 57. The formulation as described in any of paragraphs 33-56, wherein the additional active agent carrier comprises a surface modifier located on its surface.

[0331] 58. The formulation as described in any of paragraphs 33-57, wherein the surface modifier of the additional active agent carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0332] 59. The formulation as described in any of paragraphs 33-56, wherein the surface of the additional active agent carrier is substantially free of surface modifiers.

[0333] 60. The formulation as described in any of paragraphs 33-59, wherein the formulation further comprises a zinc compound.

[0334] 61. The formulation as described in any of paragraphs 33-60, wherein the formulation further comprises a humectant.

[0335] 62. A dual rational therapy (DART) molecule with two distinct antibacterial mechanisms of action.

[0336] 63. A DART molecule having: a β-lactam ring and a quinolone core; or a quinolone core and a nitro heterocycle; or a β-lactam ring and a nitro heterocycle.

[0337] 64. The molecule as described in paragraph 62, wherein the molecule inhibits DNA gyrase or topoisomerase IV and transpeptidase-mediated peptidoglycan crosslinking.

[0338] 65. The molecule as described in paragraph 62 or 63, wherein the molecule inhibits isoprenyl pyrophosphate and transpeptidase-mediated peptidoglycan crosslinking.

[0339] 66. The molecule as described in any of paragraphs 62-64, wherein the molecule inhibits isoprenyl pyrophosphate and topoisomerase IV DNA gyrase.

[0340] 67. The molecule described in any of paragraphs 62-65, wherein the molecule inhibits folic acid synthesis and DNA gyrase topoisomerase IV.

[0341] 68. The molecule as described in any of paragraphs 62-66, wherein the molecule inhibits folic acid synthesis and transpeptidase-mediated peptidoglycan crosslinking.

[0342] 69. The molecule described in any of paragraphs 62-67, wherein the molecule inhibits DNA gyrase or topoisomerase IV and the 30S subunit in bacteria.

[0343] 70. The molecule described in any of paragraphs 62-68, wherein the molecule inhibits DNA gyrase or topoisomerase IV and the 50S subunit in bacteria.

[0344] 71. The molecule as described in any of paragraphs 62-69, wherein the molecule inhibits transpeptidase-mediated peptidoglycan crosslinking and the 30S or 50S subunit in bacteria.

[0345] 72. The molecule as described in any of paragraphs 62-70, wherein the molecule inhibits folic acid synthesis and the 30S or 50S subunit in bacteria.

[0346] 73. The molecule as described in any of paragraphs 62-71, wherein the molecule inhibits isoprenyl pyrophosphate and the 30S or 50S subunit in bacteria.

[0347] 74. A dual rational therapy (DART) molecule with two distinct mechanisms of action against acne.

[0348] 75. The molecule as described in paragraph 73, wherein the molecule modulates at least two different targets.

[0349] 76. The molecule as described in paragraph 73 or 74, wherein the first mechanism of action is antibacterial action and the second mechanism of action is inhibition of keratinocyte proliferation and differentiation.

[0350] 77. The molecule as described in any one of paragraphs 73-75, wherein the first mechanism of action is antibacterial action and the second mechanism of action is anti-inflammatory action.

[0351] 78. A dual rational therapy (DART) molecule comprising two chemical domains, each of which binds to a distinct active site in a target cell, wherein the two chemical domains are linked together by a third domain.

[0352] 79. The molecule as described in paragraph 77, wherein the third structural domain is a connector.

[0353] 80. The molecule as described in paragraph 77 or 78, wherein the third structural domain is a breakable connector.

[0354] 81. The molecule as described in paragraph 77 or 78, wherein the third structural domain is an unbreakable connector.

[0355] 82. The molecule as described in any of paragraphs 77-80, wherein the third structural domain is 11-hydroxyundecenoic acid, 1,10-decanediol, 1,3-propanediol, 1,5-pentanediol, 10-hydroxydecenoic acid, succinic acid, lactic acid, 3-hydroxypropionic acid, or any combination thereof.

[0356] 83. The molecule as described in any of paragraphs 77-81, wherein the third domain increases the activity of at least one of the two chemical domains.

[0357] 84. The molecule as described in any of paragraphs 77-82, wherein the third domain has antibacterial or anti-inflammatory activity.

[0358] 85. The molecule as described in any of paragraphs 62-83, wherein the molecule is in the form of a drug carrier.

[0359] 86. The molecule as described in any of paragraphs 62-84, wherein the size of the drug carrier is from about 5 μm to about 100 μm.

[0360] 87. The molecule as described in any of paragraphs 62-85, wherein the size of the drug carrier is from about 100 nm to about 25 μm.

[0361] 88. The molecule as described in any of paragraphs 62-86, wherein the drug carrier further comprises a compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0362] 89. The molecule as described in any of paragraphs 62-87, wherein the drug carrier further comprises an additional active agent.

[0363] 90. The molecule as described in paragraph 88, wherein the additional active agent is an anti-inflammatory agent, a stratum corneum separator, a penetration enhancer, an antioxidant, an anti-aging agent, an anti-wrinkle agent, a skin brightener or bleaching agent, an ultraviolet (UV) absorber or scattering agent, a skin depigmenting agent, a skin regenerating agent, a scar healing agent, or any combination thereof.

[0364] 91. The molecule as described in any of paragraphs 62-89, wherein the surface of the drug carrier is substantially free of surface modifiers.

[0365] 92. The molecule as described in any of paragraphs 62-90, wherein the drug carrier further comprises an additional anti-acne agent.

[0366] 93. The molecule as described in any of paragraphs 62-91, wherein the second anti-acne agent is selected from the group consisting of: acitretin, adapalene, retinoic acid, α-hydroxy acid or β-hydroxy acid, antibiotics, antimicrobial peptides, antimicrobial agents, azelaic acid, benzoyl peroxide, bexarotine, bile salts, biofilm inhibitors, clindamycin, erythromycin, etretinate, glycolic acid, isotretinoin, keratolytic agents, lactic acid, lipoic acid, N-acetylcysteine, natural anti-acne agents, oxymethopyrone, phenoxyethanol, phenoxypropanol, pyruvate, resorcinol, retinoic acid, retinoids, salicylic acid, sebostats, sodium sulfacetamide, spironolactone, sulfur, sulfur-containing D-amino acids or L-amino acids, tazarotene, tea tree oil, retinoic acid, triclosan, urea, and any combination thereof.

[0367] 94. The molecule as described in any of paragraphs 62-92, wherein the drug carrier further comprises a surface modifier located on its surface.

[0368] 95. The molecule as described in any of paragraphs 62-93, wherein the surface modifier is a compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0369] 96. A formulation comprising a dual-effect rational therapy molecule as described in any of paragraphs 62-94 and at least one carrier or excipient.

[0370] 97. The formulation as described in paragraph 95, wherein the carrier or excipient is selected from the group consisting of: emulsifiers, preservatives, surfactants, oils, fats, waxes, stabilizers, rheology modifiers or thickeners (gelling agents), emollients, humectants, conditioning agents, fragrances / perfumes, synergists, preservatives, opacifiers, antioxidants, cooling agents, film-forming agents, exfoliants, scabies, colorants, pH adjusters, solvents, solvents, penetration enhancers, permeation enhancers, pearlescent agents, and any combination thereof.

[0371] 98. The formulation as described in paragraph 95 or 96, wherein the formulation comprises about 5% to about 99% (w / w or w / v) of the carrier or excipient.

[0372] 99. The formulation as described in any of paragraphs 95-97, wherein the formulation is formulated for topical administration, oral administration or parenteral administration.

[0373] 100. The formulation as described in any of paragraphs 95-98, wherein the formulation is an oral dosage form, an injection, an aerosol or inhaler, a lotion, a cream, a gel, a latex, an oil, a serum, a powder, a spray, an ointment, a solution, a suspension, a dispersant, a paste, a foam, a release agent, a film, a mask, a patch, a stick, a roll-on, a fabric soaked in fabric (e.g., a “rag” or a paper towel), or any combination thereof.

[0374] 101. The formulation as described in any of paragraphs 95-99, wherein the formulation further comprises a second anti-acne agent.

[0375] 102. The formulation as described in any of paragraphs 95-100, wherein the second antibacterial agent is selected from the group consisting of: acitretin, adapalene, ivermectin, α-hydroxy acids or β-hydroxy acids, antibiotics, antimicrobial peptides, antimicrobial agents, azelaic acid, benzoyl peroxide, bexarotine, bile salts, biofilm inhibitors, clindamycin, erythromycin, etretinate, glycolic acid, isotretinoin, keratolytic agents, lactic acid, lipoic acid, N-acetylcysteine, natural anti-acne agents, oxymethopyrone, phenoxyethanol, phenoxypropanol, pyruvate, resorcinol, retinoic acid, retinoids, salicylic acid, sebostats, sodium sulfacetamide, spironolactone, sulfur, sulfur-containing D-amino acids or L-amino acids, tazarotene, tea tree oil, retinoic acid, triclosan, urea, and any combination thereof.

[0376] 103. The formulation as described in any of paragraphs 95-101, wherein the second anti-acne agent is in the form of a drug delivery vehicle.

[0377] 104. The formulation as described in any of paragraphs 95-102, wherein the second anti-acne agent carrier further comprises a compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0378] 105. The formulation as described in any of paragraphs 95-103, wherein the size of the second anti-acne agent carrier is from about 5 nm to about 100 μm.

[0379] 106. The formulation as described in any of paragraphs 94-103, wherein the size of the second anti-acne agent carrier is from about 100 nm to about 25 μm.

[0380] 107. The formulation as described in any of paragraphs 95-105, wherein the second anti-acne agent carrier comprises a surface modifier located on its surface.

[0381] 108. The formulation as described in any of paragraphs 95-106, wherein the surface modifier of the second anti-acne agent carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0382] 109. The formulation as described in any of paragraphs 95-105, wherein the surface of the second anti-acne agent is substantially free of surface modifiers.

[0383] 110. The formulation as described in any of paragraphs 95-108, wherein the formulation further comprises an additional active agent.

[0384] 111. The formulation as described in any of paragraphs 95-109, wherein the additional active agent is an anti-inflammatory agent, a penetration enhancer, an antioxidant, an anti-aging agent, an anti-wrinkle agent, a skin brightener or bleaching agent, an ultraviolet (UV) absorber or scattering agent, a skin depigmenting agent, a skin regenerating agent, a scar healing agent, or any combination thereof.

[0385] 112. The formulation as described in any of paragraphs 95-110, wherein the additional active agent is in the form of a drug carrier.

[0386] 113. The formulation as described in any of paragraphs 95-111, wherein the additional active agent carrier further comprises a compound selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0387] 114. The formulation as described in any of paragraphs 95-112, wherein the size of the additional active agent carrier is from about 5 nm to about 100 μm.

[0388] 115. The formulation as described in any of paragraphs 95-113, wherein the size of the additional active agent carrier is from about 100 nm to about 25 μm.

[0389] 116. The formulation as described in any of paragraphs 95-114, wherein the additional active agent carrier comprises a surface modifier located on its surface.

[0390] 117. The formulation as described in any of paragraphs 95-115, wherein the surface modifier of the additional active agent carrier is selected from the group consisting of lipids, oils, polymers, peptides, proteins, carbohydrates, glycolipids, phospholipids, lipoproteins, cationic molecules, and any combination thereof.

[0391] 118. The formulation as described in any of paragraphs 95-114, wherein the surface of the additional active agent carrier is substantially free of surface modifiers.

[0392] 119. The formulation as described in any of paragraphs 95-117, wherein the formulation further comprises a zinc compound.

[0393] 120. A method for treating acne in a subject, the method comprising administering a therapeutically effective amount of the preparation described in any one of paragraphs 1-61 and 95-118.

[0394] 121. The method as described in paragraph 119, wherein the acne condition is caused by an antibiotic-sensitive bacterial strain.

[0395] 122. The method as described in paragraphs 119 or 120, wherein the acne condition is caused by antibiotic-resistant bacteria.

[0396] 123. The method described in any of paragraphs 119-121, wherein the acne condition is caused by clindamycin, tetracycline, doxycycline or erythromycin-resistant Propionibacterium acnes.

[0397] 124. The method described in any paragraph 119-122, wherein the acne condition is caused by clindamycin, tetracycline, doxycycline or erythromycin-resistant Propionibacterium acnes.

[0398] 125. A method for treating a bacterial infection in a subject, the method comprising administering a therapeutically effective amount of a preparation as described in any one of paragraphs 1-61 and 95-118.

[0399] 126. The method as described in paragraph 124, wherein the infection is caused by a pathogen selected from the group consisting of Bartonella henselae (…). Bartonella henselae Borrelia burgdorferi () Borrelia burgdorferi Campylobacter jejuni ( Campylobacter jejuni Campylobacter fetus ( Campylobacter fetus ), Chlamydia trachomatis ( Chlamydia trachomatis Chlamydia pneumoniae ( Chlamydia pneumonia ), Chlamydia psittaci ( Chylamydia psittaci ), Simkania negevensis Escherichia coli ( Escherichia coli (e.g., O157:H7 and K88), Ericksonites ( Ehrlichia chafeensis Clostridium botulinum ( Clostridium botulinum Clostridium perfringens ( ) Clostridium perfringens Clostridium tetani ( Clostridium tetani ), Enterococcus faecalis ( Enterococcus faecalis Haemophilus influenzae ( ) Haemophilus influenzae ), Haemophilus ducreyi ( Haemophilius ducreyi ), Coccidioides immitis ( Coccidioides immitis Bordetella pertussis ( ) Bordetella pertussis ), Rickettsia burnetii ( Coxiella burnetii ), Ureaplasma urealyticum ( Ureaplasma urealyticum ), Mycoplasma genitalium ( Mycoplasma genitals ), Trichomonas vaginalis Helicobacter pylori ( Helicobacter pylori ), Helicobacter hepatis ( Helicobacter hepaticus Legionella pneumophila ( Legionella pneumophila ), Mycobacterium tuberculosis ( Mycobacterium tuberculosis), Mycobacterium bovis ( Mycobacterium bovis ), Mycobacterium africanum ( Mycobacterium africanum Mycobacterium leprae ( Mycobacterium leprae ), Mycobacterium Asiatum ( Mycobacterium asiaticum ), Mycobacterium avium ( Mycobacterium avium ), Mycobacterium cryptans ( Mycobacterium cryptatum ), Mycobacterium tectorum ( Mycobacterium chelonae Occasionally occurring mycobacteria ( Mycobacterium fortuitum ), Mycobacterium Genevai ( Mycobacterium genavense Haemophilus ( ) Mycobacterium haemophilus Intracellular mycobacteria ( Mycobacterium intracellulare ), Mycobacterium Kansas ( Mycobacterium kansasii ), Mycobacterium marmosetum ( Mycobacterium malmoensis ), Mycobacterium marinum ( Marine Mycobacterium ), Mycobacterium scrofula ( Mycobacterium scrofulaceum Mycobacterium simianum ( Mycobacterium simianum ), Mycobacterium stearothermia ( Mycobacterium szulgai ), Mycobacterium ulcerans ( Mycobacterium ulcerans ), Mycobacterium bufossa ( Mycobacterium xenopi Corynebacterium diphtheriae ( Corynebacterium diptheriae ), Rhodococcus equi ( Rhodococcus equi ), Rickettsia ehrlichii ( Rickettsia aeschlimannii ), African rickettsia ( Rickettsia africae ), Connorricklet ( Rickettsia conorii Cryptolytica hemolyticus ( Arcanobacterium haemolyticum Bacillus anthracis ( Bacillus anthracis ), Bacillus cereus ( Bacillus cereus Listeria monocytogenes ( ) Lysteria monocytogenes Yersinia pestis (Yersinia pestis) Yersinia pestis Yersinia enterocolitica (Yersinia enterocolitica) Yersinia enterocolitica ), Shigella dysenteriae ( Shigella dysenteriae ), Neisseria meningitidis ( Neisseria meningitides ), Neisseria gonorrhoeae ( Neisseria gonorrhoeae Streptococcus bovis () Streptococcus bovis ), hemolytic streptococci ( Streptococcus hemolyticus Streptococcus mutans () Streptococcus mutans Streptococcus pyogenes ( ) Streptococcus pyogenes Streptococcus pneumoniae () Streptococcus pneumoniae Staphylococcus aureus ( Staphylococcus aureus Staphylococcus epidermidis ( Staphylococcus epidermidis Staphylococcus pneumoniae () Staphylococcus pneumoniae ), saprophytic Staphylococcus ( Staphylococcus saprophyticus ), Vibrio cholerae ( Vibrio cholerae ), Vibrio parahaemolyticus ( Vibrio parahaemolyticus ), Salmonella typhi Salmonella typhi Salmonella paratyphi ()Salmonella paratyphi Salmonella enteritidis ( ) Salmonella enteritidis Treponema pallidum (Syphilis) Treponema pallidum Candida ( Candida Cryptococcus ( Cryptcooccus Cryptosporidium ( Cryptosporidium Giardia lamblia ( Giardia lamblia Microsporidia ( Microsporidia ), Plasmodium vivax ( Plasmodium vivax Pneumocystis carinii ( Pneumocystis carinii ), Toxoplasma gondii ( Toxoplasma gondii ), Trichophyton mentagrophytes ( Trichophyton mentagrophytes ), Intestinal microsporidia ( Enterocytozoon bieneusi Cyclospora ( Cyclospora cayetanensis ), Helen's encephalitis microsporidia ( Encephalitozoon hellem Rabbit intracellular protozoa ( Encephalitozoon cuniculi ), as well as other bacteria, archaea, protozoa and fungi.

[0400] 127. The method as described in paragraphs 124 or 125, wherein the infection is caused by an antibiotic-resistant bacterial strain.

[0401] 128. The method as described in paragraphs 124 or 125, wherein the infection is caused by an antibiotic-sensitive bacterial strain.

[0402] 129. The method as described in any of paragraphs 124-127, wherein the formulation is administered to the subject once or daily in a single or multiple doses.

[0403] Some selected definitions

[0404] For convenience, certain terms used in this specification, embodiments, and appended claims are collected herein. Unless otherwise stated or implied by the context, the following terms and phrases have the meanings provided below. Unless otherwise expressly stated or clearly apparent from the context, the following terms and phrases do not exclude the meanings already obtained in the art to which they pertain. Definitions are provided to aid in the description of specific embodiments and are not intended to limit the claimed invention, as the scope of the invention is defined only by the claims. Furthermore, unless the context requires otherwise, singular terms shall include plurals and plural terms shall include singulars.

[0405] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any known methods, apparatus, and materials may be used in the practice or testing of this invention, for this purpose they are described herein.

[0406] As used herein, the term “this article” refers to the entire public text and therefore does not imply limitation to any particular area or sub-area of ​​this public text.

[0407] As used herein, the term "comprising / including" refers to compositions, methods, and their respective components that are essential to the method or composition, while remaining open to unspecified elements, whether or not such unspecified elements are essential.

[0408] Unless the context clearly indicates otherwise, the singular terms “a / an / the” include a plural number of objects. Similarly, unless the context clearly indicates otherwise, the word “or” is intended to include “and”.

[0409] Except where indicated in the operational embodiments or elsewhere, all numerical values ​​representing quantities of ingredients or reaction conditions as used herein should in all cases be understood to be modified by the term “about”. The term “about” used in conjunction with percentages may mean ±5%, ±4%, ±3%, ±2.5%, ±2%, ±1.5%, ±1%, or ±0.5% of the value mentioned.

[0410] While methods and materials similar to or equivalent to those described and used herein may be used in practice or testing of this disclosure, suitable methods and materials are described below. The term “comprising” means “containing”. The abbreviation “eg” comes from the Latin word *exempli gratia* and is used herein to indicate a non-limiting instance. Therefore, the abbreviation “eg” is a synonym for the term “for example”.

[0411] As used herein, the terms “decrease / reduced / reduction / decrease / inhibit” generally mean a statistically significant reduction. However, to avoid any ambiguity, “decrease / reduced / inhibit” means a reduction of at least 10% compared to a reference level, such as 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 100% reduction (e.g., a level that does not exist compared to a reference instance), or any reduction between 10% and 100% compared to a reference level.

[0412] As used in this document, the terms "increased / increase / enhance / active" generally mean a statistically significant increase. For the avoidance of any ambiguity, the terms "increased / increase / enhance / active" mean an increase of at least 10% compared to a reference level, such as 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 100% increase, or any increase between 10% and 100% compared to a reference level, or an increase of at least about 2 times, or at least about 3 times, or at least about 4 times, or at least about 5 times, or at least about 10 times compared to a reference level, or any increase between 2 times and 10 times or higher compared to a reference level.

[0413] The terms "statistically significant" or "significant" refer to statistical significance and generally mean a deviation from a reference level of at least two standard deviations (2SD). This term refers to statistical evidence indicating a difference. It is defined as the probability of making a decision to reject the null hypothesis when it is actually true.

[0414] As used herein, the term "sphere" refers to a spherical or quasi-spherical object, a ball, or other shape of particle of a substance (e.g., in the form of a two-phase suspension or emulsion). Subdivisions of solid materials are also included in the meaning of the term "sphere."

[0415] The present disclosure is further illustrated by the following examples, which should not be construed as limiting. The examples are merely illustrative and are not intended to limit any aspect described herein in any way. The following examples do not limit the invention in any way.

[0416] Example

[0417] Example 1: Screening for antibiotics against Propionibacterium acnes strains showed that the response was unpredictable in both clindamycin-sensitive and non-responsive strains.

[0418] Examples 2-15 and Tables 6-15 describe some exemplary DART molecules, their synthesis, formulation, and uses.

[0419] Example 2: Synthesis of DART molecule 9 from Table 1A

[0420]

[0421] Synthesis of Steps 1 and 2: N,N-dicyclohexylcarbodiimide (0.627 g, 3.041 mmol) was added to a solution of 1 (1 g, 2.33 mmol) in a mixture of dichloromethane (10 mL) and dimethylformamide (1 mL), followed by the slow addition of N-hydroxybenzotriazole (0.316 g, 2.33 mmol) under ice-cold conditions, and the mixture was stirred at room temperature for 2 hours to obtain a turbid suspension. Pentylene glycol (0.85 mL, 8.18 mmol) was added to this turbid solution, followed by 4-dimethylaminopyridine (0.284 g, 2.33 mmol). The final reaction mixture was stirred at room temperature for 16 hours. The white precipitate was filtered and extracted with ethyl acetate. The filtrate was washed with brine, dried over sodium sulfate, and evaporated to give the crude product. The crude product was purified by rapid column chromatography, eluting with 1% methanol / dichloromethane, to give pure compound 2 (0.9 g, 80% yield).

[0422] Step 2, Synthesis of DART9: Dicyclohexylcarbodiimide (0.415 g, 2.05 mmol) was added to a solution of compound 3 (0.72 g, 1.55 mmol) in a mixture of dichloromethane (10 ml) and dimethylformamide (1 ml), followed by the addition of N-hydroxybenzotriazole (0.209 g, 1.55 mmol) at room temperature, resulting in a turbid suspension. The reaction mixture was stirred at room temperature for 3 hours, and compound 2 (0.79 g, 1.55 mmol) was added to the turbid solution, followed by the addition of 4-dimethylaminopyridine (0.189 g, 1.55 mmol). The final solution was stirred at room temperature for 16 hours. The precipitate was filtered, and the filtrate was extracted with ethyl acetate. The organic layer was washed with aqueous brine and dried over sodium sulfate to give the crude product. The crude product was purified by rapid column chromatography to obtain the final product (9) in 50%–60% fraction.

[0423] Example 2: Synthesis of DART molecule 87 from Table 1A

[0424]

[0425] Synthesis of steps 1 and 4: 11-Bromoundecanoic acid (1.33 g, 5.04 mmol) was premixed with methanol (0.1 mL) and added to a stirred mixture of compound 1 (1 g, 2.52 mmol), potassium carbonate (0.243 g, 1.764 mmol), and dipotassium hydrogen phosphate (0.175 g, 1 mmol) in N,N-dimethylacetamide (15 mL) at 0–5 °C. The reaction mixture was stirred at 0 °C for 5 hours and extracted with ethyl acetate (50 mL). The final solution was washed with 3% sodium bicarbonate aqueous solution (10 mL), followed by a brine solution (10 mL). The organic solvent was evaporated to give crude material, which was then purified by rapid column chromatography. The compound was eluted with 1%–3% methanol / dichloromethane to give pure compound 4 (1.2 g, 82% yield).

[0426] Step 2, Synthesis of DART87: Dicyclohexylcarbodiimide (0.461 g, 2.23 mmol) was added to a solution of compound 4 (1 g, 1.72 mmol) in dichloromethane (10 ml) and dimethylformamide (1 ml), followed by the addition of N-hydroxybenzotriazole (0.232 g, 1.72 mmol) at room temperature, resulting in a turbid suspension. The reaction mixture was stirred at room temperature for 1 hour. Compound 5 (0.67 g, 1.72 mmol) was added to the turbid solution, followed by the addition of DMAP (0.210 g, 1.72 mmol), and the reaction mixture was stirred at room temperature for 16 hours. The suspension was filtered and washed with a saline solution. The organic layer was dried over sodium sulfate and evaporated to obtain the crude product. The final crude product was purified by rapid column chromatography using 2%–5% methanol / dichloromethane as eluent, yielding pure compound 87 in 60%–65% separation yield.

[0427] Example 3: Synthesis of DART molecule 90 from Table 1B

[0428] The following scheme was used to synthesize 1-chloromethyl-2-(2-methyl-5-nitroimidazol-1-yl)-ethyl bromoacetic acid (II).

[0429]

[0430] Note: Compound 1 in the scheme corresponds to compound 90 in Table 1B.

[0431] To a stirred solution of 1-chloro-3-(2-methyl-5-nitroimidazol-1-yl)-propane-2-ol (I) (0.79 g, 3.6 mmol) in dichloromethane (10 mL), dicyclohexylcarbodiimide (DCC) (0.9 g, 4.31 mmol) was added, followed by the addition of bromoacetic acid (0.5 g, 3.6 mmol) and DMAP (0.44 g, 3.6 mmol) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The precipitate was removed by filtration and the organic layer was evaporated to give the crude product, which was purified by rapid column chromatography. The final compound was eluted with a 1%–2% methanol / dichloromethane mixture. The compound was used in the next step without further characterization.

[0432] Synthesis of 7-{4-[1-chloromethyl-2-(2-methyl-5-nitroimidazol-1-yl)-ethoxycarbonylmethyl]-piperazin-1-yl}-6-fluoro-1-methyl-4-oxo-4H-2-thia-8b-aza-cyclobutane[a]naphthalene-3-carboxylic acid (1): Potassium carbonate (0.04 g, 0.3 mmol) was added to a stirred solution of 6-fluoro-1-methyl-4-oxo-7-piperazin-1-yl-4H-2-thia-8b-aza-cyclobutane[a]naphthalene-3-carboxylic acid (III) (0.071 g, 0.2 mmol) in dimethylformamide (10 ml), followed by compound (II) (0.1 g, 0.3 mmol). The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with ethyl acetate, washed twice with water, and finally dried with sodium sulfate to give the crude substance. The crude product was purified by rapid column chromatography, eluted with a 3%-5% methanol / dichloromethane mixture, and the pure compound (1), i.e., compound 90 in Table 1, was obtained in a 30% separation yield.

[0433] 1 H-NMR (400 MHz, DMSO) ppm: 2.19 (3H, d, J = 6.4 Hz, CH3), 2.57 (3H, s,CH3), 2.7 (4H, m, 2×CH2), 3.1-3.3 (2H, s, COCH2), 3.32 (4H, m, 2×CH2), 3.77-3.90(2H, ddd, J 1 = 3.6 Hz J 2 = 12.4 Hz J 3= 35.2 Hz, CH2Cl), 4.40-4.56 (1H, dd, J 1 = 9.6 Hz J 2 = 14 Hz CHN), 4.76 (1H, d, J= 14 Hz, CHN), 5.44 (1H, d, J =5.6HzCHOCO), 6.0-6.11 (1H, q, J 1 = 6 Hz J 2= ​​12.4 Hz, CHSN), , 6.4 (1H, d, J 1 = 6.8 Hz Ar-H), 7.8 (1H, d, J = 14 Hz, Ar-H), 8.04 (1H, s, Ar-H). ESI-MS (m / z): 609 (M+H) + .

[0434] Example 4: Synthesis of DART molecule 91 from Table 1B

[0435] 2-Methyl-5-nitro-1-epoxyethylenemethyl-1H-imidazolium (IV) was synthesized according to the following scheme.

[0436]

[0437] Note: Compound 2 in the scheme corresponds to compound 91 in Table 1B.

[0438] At 0 °C, 20% sodium hydroxide (4 ml) was added to a stirred solution of 0.5 g (2.27 mmol) of 1-chloro-3-(2-methyl-5-nitroimidazol-1-yl)-propane-2-ol (I) in 8 ml of dichloromethane. The reaction mixture was stirred at 0 °C for 3 hours. After 3 hours, the reaction mixture was extracted twice with dichloromethane, the organic layers were combined, washed with brine, and finally dried over sodium sulfate to give the pure product in 90% separation yield. 1 H-NMR (400 MHz, CDCl3) ppm: 2.517 (3H, s,CH3), 2.52 (1H, m, CH2), 2.88 (1H, m, CH2), 3.38 (1H, m, CH), 4.17-4.23 (1H,dd, J 1 = 6Hz J 2= ​​15.2Hz CH2), 4.85-4.89 (1H, , d , J = 14.8 CH2).

[0439] Synthesis of 6-fluoro-7-{4-[2-hydroxy-3-(2-methyl-5-nitroimidazol-1-yl)-propyl]-piperazin-1-yl}-1-methyl-4-oxo-4H-2-thia-8b-aza-cyclobut[a]naphthalene-3-carboxylic acid (2): Potassium carbonate (0.11 g, 0.19 mmol) dissolved in water (5 ml) was added to a stirred solution of 6-fluoro-1-methyl-4-oxo-7-piperazin-1-yl-4H-2-thia-8b-aza-cyclobut[a]naphthalene-3-carboxylic acid (III) (0.2 g, 0.57 mmol) in acetone (15 ml), followed by the addition of ornidazole epoxide (II) (0.15 g, 0.82 mmol). The reaction mixture was heated at 50 °C for 20 hours. After completion, the reaction mixture was evaporated and extracted twice with dichloromethane. The combined organic layers were dried over sodium sulfate and evaporated to give the crude substance. The crude substance was purified by column chromatography, eluted with a 10%-12% methanol / dichloromethane mixture, and the pure compound (2), namely, compound 91 in Table 1, was obtained with a separation yield of 25%-30%. 1 H-NMR (400 MHz, DMSO) δppm: 2.12 (3H, d, J = 6.4 Hz, CH3), 2.46(3H, s, CH3), 2.58-2.60 (2H, t, J = 5.6 Hz CH2N ), 2.66 (4H, m, 2×CH2), 3.2(4H, m, 2 x CH2), 3.9-4.1 (2H, m, 2×CH2N), 4.63 (1H, d, J = 14 Hz, CHOH), 5.15(1H, d, J = 5.2 Hz -OH), 6.39 (1H, d, J = 6.4 Hz, CHSN) 6.93 (1H, d, J = 7.2 Hz (Ar-H), 7.79 (1H, d, J = 14 Hz, Ar-H), 8.04 (1H, s, Ar-H). ESI-MS (m / z): 532.95(M+H).

[0440] Example 5: Synthesis of DART molecule 94 from Table 1B

[0441]

[0442] Note: Scheme 5 corresponds to compound 94 in Table 1B.

[0443] Synthesis of 6-fluoro-1-methyl-7-[4-(5-methyl-2-oxo-[1,3]dioxo-4-ylmethyl)-piperazin-1-yl]-4-oxo-4H-2-thia-8b-aza-cyclobutane-3-carboxylic acid 1-chloromethyl-2-(2-methyl-5-nitroimidazol-1-yl)-ethyl ester (5): To 6-fluoro-1-methyl-7-[4-(5-methyl-2-oxo-[1,3]dioxocyclopenten-4-ylmethyl)-piperazin-1-yl]-4-oxo-4H-2- DCC (0.3 g, 1.41 mmol) and HOBt (0.146 g, 1.083 mmol) were added to a stirred solution of thia-8b-aza-cyclobutane[a]naphthalene-3-carboxylic acid (V) (0.5 g, 1.08 mmol) in DMF (20 mL), followed by the addition of 1-chloro-3-(2-methyl-5-nitroimidazole-1-yl)-propane-2-ol (I) (0.285 g, 1.3 mmol) and DMAP (0.13 g, 1.08 mmol) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The precipitate was removed by filtration and the organic layer was evaporated to give the crude substance. Finally, the crude substance was purified by rapid column chromatography, eluting with a 2%–4% methanol / dichloromethane mixture, to give the pure compound (5), i.e., compound 94 in Table 1, in 60% separation yield. 1 H-NMR (400 MHz, DMSO) δppm:2.03 (3H,d, J =5.6Hz, CH3), 2.12 (3H, s, CH3), 2.5 (3H, s, CH3), 2.62 (4H, m, 2×CH2), 3.22 (4H, m, 2 x CH2), 3.95-4.06 (2H, m, CH2Cl), 4.49-4.52 (1H, t, J = 10 Hz, CHN) 4.77 (1H, d, J = 13.2Hz, CHN), 5.63(1H, d, J = 4.4Hz, CHOCO), 6.15 (1H, m,CHSN), 6.78 (1H, d, J = 7.2, Ar-H), 7.68 (1H, d, J = 14, Ar-H), 7.9 (1H, s, Ar-H).

[0444] Example 6: Synthesis of DART molecule 116 from Table 1B

[0445]

[0446] Synthesis of 4-bromo-1,2-epoxybutane (I): A solution of 3-chloroperbenzoic acid (55%-75% purity, 1.60 g, 9.25 mmol) in 10 mL of dichloromethane was added dropwise to a stirred solution of 4-bromo-1-butene (0.5 g, 3.7 mmol) in 20 mL of dichloromethane. After addition, the mixture was stirred at 25 °C for 16 hours to precipitate 3-chlorobenzoic acid. Finally, the reaction mixture was evaporated to dryness under vacuum, dissolved in ethyl acetate, washed first with 4% sodium dithionate, then with saturated sodium bicarbonate and water. Finally, the organic layer was dried with sodium sulfate and evaporated to dryness under vacuum to obtain the final compound (I) in 90% isolated yield. 1 H-NMR (400 MHz, CDCl3) δppm: 2.10 (m, 2H, CH2) 2.58 (s, 1H, OCH a ) 2.82 (, 1H, m,OCH b ), 3.09 (m, 1H, CH) 3.55 (t, J = 6.4, 2H, CH2-Br).

[0447] Synthesis of 4-bromo-1-(2-methyl-5-nitroimidazol-1-yl)-butane-2-ol (II): Anhydrous aluminum chloride (1.67 g, 12.5 mmol) was added to a stirred solution of 2-methyl-5-nitro-1H-imidazol (0.8 g, 6.3 mmol) in anhydrous ethyl acetate at 0 °C and stirred for 15 min to dissolve the 2-methyl-5-nitro-1H-imidazol. 4-bromo-1,2-epoxybutane (I) (1.9 g, 12.5 mmol) was added dropwise to the reaction mixture, and the reaction mixture was continued to react at 0 °C for 5 h. The reaction mixture was slowly added to ice water, and the pH was adjusted to 1 by adding concentrated HCl. The organic layers were separated and washed with saturated sodium bicarbonate followed by water. The pH of the aqueous layer obtained from the first separation was adjusted to 7.4 using ammonia and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate and evaporated under vacuum to give the crude compound. The crude product was purified by rapid column chromatography, eluting with a 2%–3% methanol / dichloromethane mixture, to give pure compound (II) in 50% fraction. ESI-MS (m / z): 277(M) +

[0448] Synthesis of 6-fluoro-7-{4-[3-hydroxy-4-(2-methyl-5-nitroimidazol-1-yl)-butyl]-piperazin-1-yl}-1-methyl-4-oxo-4H-2-thia-8b-aza-cyclobut[a]naphthalene-3-carboxylic acid (116): Potassium carbonate (1.20 g, 8.6 mmol) was added to a stirred solution of 6-fluoro-1-methyl-4-oxo-7-piperazin-1-yl-4H-2-thia-8b-aza-cyclobut[a]naphthalene-3-carboxylic acid (III) (3 g, 8.6 mmol) in DMF (30 mL), followed by compound (II) (2 g, 7.2 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with ethyl acetate, washed twice with water, and finally dried with sodium sulfate to give the crude substance. The crude product was purified by rapid column chromatography, eluting with a 3%-5% methanol / dichloromethane mixture, to give pure compound (116) in 20% separation yield. 1 H-NMR (400 MHz, DMSO) δppm: 1.61-1.68 (2H, m, CH2),2.12 (3H, d, J = 6 Hz, CH3),2.46 (3H, s, CH3), 2.57 (4H, m, 2×CH2), 3.2 (4H, m,2×CH2), 3.8-4.1 (2H, m, 2×CH2N), 4.44(1H, m, CHOH), 5.2 (1H, bs, CHOH), 6.4(1H, q, J 1 = 5.6Hz J 2 = 11.6 Hz, CHSN) 6.91 (1H, d, J = 7, Ar-H), 7.78 (1H, d, J =13.6Hz, Ar-H), 8.04 (1H, s, Ar-H). ESI-MS (m / z): 547.08(M+H) + .

[0449] Example 7: Synthesis of DART molecule 113 from Table 1B

[0450]

[0451] Synthesis of 7-{4-[2-dodecanoyloxy-3-(2-methyl-5-nitroimidazol-1-yl)-propyl]-piperazin-1-yl}-6-fluoro-1-methyl-4-oxo-4H-2-thia-8b-aza-cyclobutane[a]naphthalene-3-carboxylic acid (113):

[0452] To a solution of 91 (0.5 g, 0.94 mmol) of dimethylformamide (15 mL), N,N-dicyclohexylcarbodiimide (0.29 g, 1.41 mmol) was added, followed by the slow addition of N-hydroxybenzotriazole (0.13 g, 0.94 mmol) under ice-cold conditions. The mixture was stirred at room temperature for 10 min to obtain a turbid suspension. Lauric acid (0.28 g, 1.5 mmol) was added to this turbid solution, followed by 4-dimethylaminopyridine (0.115 g, 0.94 mmol). The final reaction mixture was stirred at room temperature for 16 h. The white precipitate was filtered off and extracted with ethyl acetate. The filtrate was washed with water and brine, dried over sodium sulfate, and evaporated to give the crude product. The crude product was purified by rapid column chromatography, eluting with 2%–3% methanol / dichloromethane, to give pure compound 113 in 35% fraction. 1 H-NMR (400 MHz, CDCl3) δppm: 0.86(3H, t, J = 6 Hz, CH3), 1.05-1.38(18H, m, -CH2), 1.48-1.70(4H, m, -CH2), 1.90-1.93(2H, d, J = 11.6Hz, CH3),2.16-2.22 (3H, m, CH3), 2.53 (3H, s, CH3), 2.58-2.69 (2H, m, CH2N ), 2.69-2.87(4H, m, 2×CH2), 3.2-3.4 (4H, m, 2×CH2), 4.1-4.25 (2H, m, 2×CH2N), 5.0 (1H,s, CHOH), 6.09(1H, d, J = 5.2 Hz, CHSN), 6.41 (1H, d, J = 6.8 Hz Ar-H), 7.92 (1H,d, J = 14.8 Hz, Ar-H), 8.04 (1H, s, Ar-H). ESI-MS (m / z): 715.2(M+H).

[0453] Example 8: Synthesis of DART molecule 115 from Table 1B

[0454]

[0455] Synthesis of 2-(4-bromobutyl)-ethylene oxide (I): A solution of 3-chloroperbenzoic acid (55-75% purity, 4.54 g, 18.39 mmol) in 20 mL of dichloromethane was added dropwise to a stirred solution of 6-bromo-1-hexene (2 g, 12.26 mmol) in 20 mL of dichloromethane. After addition, the mixture was stirred at 25 °C for 16 hours to precipitate 3-chlorobenzoic acid. Finally, the reaction mixture was evaporated to dryness under vacuum, dissolved in ethyl acetate, washed with 4% sodium dithionite, followed by washing with saturated sodium bicarbonate and water. Finally, the organic layer was dried with sodium sulfate, evaporated, and dried under vacuum to obtain the final compound (I) in 90% isolated yield. 1 H-NMR (400 MHz, CDCl3) δ ppm: 1 H-NMR (400 MHz, CDCl3) δ ppm: 1.48-1.6(6H, m,CH2) 2.47 (1H, d, J = 2.4, OCH) 2.75 (t, J = 4, 1H, OCH b ), 2.91 (bs, 1H, OCHa)3.41 (t, J = 6.4, 2H, CH2-Br).

[0456] Synthesis of 6-bromo-1-(2-methyl-5-nitroimidazol-1-yl)-hexane-2-ol (II): Anhydrous aluminum chloride (1.46 g, 11 mmol) was added to a stirred solution of 2-methyl-5-nitro-1H-imidazol (0.7 g, 5.5 mmol) in anhydrous ethyl acetate at 0 °C, and the mixture was stirred for 15 min to dissolve the 2-methyl-5-nitro-1H-imidazol. After adding 6-bromo-1,2-epoxyhexane (I) (1.96 g, 11.02 mmol) dropwise to the reaction mixture, the reaction was continued at 0 °C for 5 h. The reaction mixture was slowly added to ice water, and the pH was adjusted to 1 with concentrated HCl. The organic layers were separated, washed with saturated sodium bicarbonate, and then washed with water. The aqueous layer obtained from the first separation was adjusted to pH 7.4 with ammonia and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate and evaporated under vacuum to give the crude compound. The crude product was purified by rapid column chromatography, eluting with a 2%–3% methanol / dichloromethane mixture, to give pure compound (II) in 50% fraction. ESI-MS (m / z): 305.95 (M+H)

[0457] Synthesis of 6-fluoro-7-{4-[5-hydroxy-6-(2-methyl-5-nitroimidazol-1-yl)-hexyl]-piperazin-1-yl}-1-methyl-4-oxo-4H-2-thia-8β-aza-cyclobutane[a]naphthalene-3-carboxylic acid (115): Potassium carbonate (0.43 g, 3.16 mmol) was added to a stirred solution of 6-fluoro-1-methyl-4-oxo-7-piperazin-1-yl-4H-2-thia-8β-aza-cyclobutane[a]naphthalene-3-carboxylic acid (III) (1.10 g, 3.16 mmol) in dimethylformamide (30 ml), followed by compound (II) (0.85 g, 2.63 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with ethyl acetate, washed twice with water, and finally dried over sodium sulfate to give the crude substance. The crude product was purified by rapid column chromatography, eluting with a 3%-5% methanol / dichloromethane mixture, to give the pure compound (115) in a 20% separation yield. 1 H-NMR (400 MHz, DMSO) δppm: 1.61-1.68(6H, m, CH2), 2.1 (3H, d, J = 6 Hz, CH3), 2.44 (3H, s, CH3) , 2.54(4H, m, 2×CH2), 3.2 (4H, m, 2×CH2), 3.9-4.1 (2H, m, 2×CH2N), 4.38(1H, d, J =14, CHOH), 5.2 (1H, d, J = 4.4, OH), 6.38 (1H, d, J = 5.6Hz, CHSN) 6.9 (1H, d, J =6.8,Ar-H), 7.78 (1H, d, J = 14 Hz, Ar-H), 8.02 (1H, s, Ar-H). ESI-MS (m / z): 575(M+H)

[0458] Example 9: Synthesis of DART molecule 119 from Table 1B

[0459]

[0460] Synthesis of 2-(2-methyl-5-nitro-1H-imidazol-1-yl)ethyl 4-methylbenzenesulfonate (I): At 0 °C, triethylamine (7.3 mL), 4-toluenesulfonyl chloride (6.42 g, 33.72 mmol), and then 4-dimethylaminopyridine (0.2 g, 1.68 mmol) were added to a stirred solution of 2-(2-methyl-5-nitroimidazol-1-yl)-ethanol (4 g, 16.86 mmol) in dichloromethane (50 mL). The reaction mixture was stirred at room temperature for 3 hours. After stirring, the reaction mixture was washed with water, 5% hydrochloric acid, saturated NaHCO3, and water. The organic layer was dried over sodium sulfate and evaporated to give the crude product. The crude product was purified by rapid column chromatography, eluting with 2%–3% methanol / dichloromethane, to obtain the pure compound in 90% fraction. 1 H-NMR (400 MHz, CDCl3)δppm: 2.45 (3H, s, Ar-CH3), 2.51(3H, s, -CH3), 4.37(2H, d, J = 4.8Hz, CH2), 4.54(2H, d, J = 4.8Hz, CH2), 7.29(2H, d, J = 8.4Hz, ArH), 7.60(2H, d, J = 8.4Hz, ArH),7.81(1H, s, ArH).

[0461] Synthesis of 6-fluoro-1-methyl-7-{4-[2-(2-methyl-5-nitroimidazol-1-yl)-ethyl]-piperazin-1-yl}-4-oxo-4H-2-thia-8b-aza-cyclobut[a]naphthalene-3-carboxylic acid (119): Potassium carbonate (2.03 g, 14.76 mmol) was added to a stirred solution of 6-fluoro-1-methyl-4-oxo-7-piperazin-1-yl-4H-2-thia-8b-aza-cyclobut[a]naphthalene-3-carboxylic acid (II) (5.16 g, 14.76 mmol) in dimethylformamide (100 ml), followed by compound (I) (2 g, 7.2 mmol). The reaction mixture was heated at 90 °C for 16 hours. The reaction mixture was diluted with ethyl acetate, washed twice with water, and finally dried with sodium sulfate to give the crude substance. The crude product was purified by rapid column chromatography, eluting with a 4%-5% methanol / dichloromethane mixture, to give the pure compound (119) in a 20% separation yield. 1 H-NMR (400 MHz, CDCl3) δppm: 2.12 (3H,d, J= 6.4 Hz, CH3), 2.52 (3H, s, CH3), 2.64 (4H, m, 2×CH2), 2.72(2H, t, J = 6Hz, CH2), 3.11-3.18 (4H, m, 2×CH2), 4.43-4.49 (2H, m, CH2N), 5.8-5.9 (1H, q, J 1 = 6.4Hz J 2 = 12.8 Hz, CHSN), 6.3 (1H, d, J = 6.8Hz, Ar-H), 7.92 (1H, s, Ar-H),7.95 (1H, s, Ar-H). ESI-MS (m / z): 503 (M+H)

[0462] Example 10: Antimicrobial susceptibility of clindamycin-sensitive and clindamycin-resistant Propionibacterium acnes

[0463] The antimicrobial susceptibility of Propionibacterium acnes was determined by the following broth dilution method.

[0464] Methods: Propionibacterium acnes (MTCC 1951 and CCARM 9010) were cultured in Brain Heart Infusion Agar (BHIA) for 48 hours under anaerobic conditions at 37°C. For MIC testing, 100 μl of BHI fermentation broth was added to all 96 wells, and 100 μl of fermentation broth containing different concentrations of cephalosporins, cefoxitin, prulifloxacin, naflufloxacin, roxithromycin, clindamycin, and besifloxacin was added to the first well of each row (1A to 1H), and serial (double) dilutions were performed up to 10 wells (columns 1 to 10 of the 96-well plate). For bacterial inoculation, the turbidity of the Propionibacterium acnes culture was adjusted to 0.5 McFarland standard (approximately 1.5 × 10⁻⁶). 8 The solution was further diluted (100-fold with sterile BHI fermentation broth). 100 μl of the diluted Propionibacterium acnes suspension was added to each well except for the sterile control well (column 12 of a 96-well plate). The inoculated plate was incubated anaerobically at 37°C for 72 hours. After incubation, the MIC was determined by adding Alamar blue dye.

[0465] Results: MIC results for clindamycin-sensitive Propionibacterium acnes strains indicated that this strain was susceptible to infection with all antibiotics. Figure 1A Interestingly, clindamycin-resistant strains also exhibit resistance to macrolides and roxithromycin. Figure 1B ).

[0466] Example 11: Determination of minimum inhibitory concentration (MIC) and dose-response curves of compounds 90, 91, 94, 113, 115 and 116 in clindamycin-sensitive (MTCC 1951) and clindamycin-unresponsive (CCARM 9010) Propionibacterium acnes strains and laboratory Staphylococcus aureus strains.

[0467] Materials: Brain heart extract fermentation broth, Propionibacterium acnes strains (MTCC 1951 and CCARM 9010), Staphylococcus aureus MTCC 6908, 96-well plates, autoclave, incubator, anaerobic chamber with anaerobic gas pack, plate reader (595nm), Alamar blue.

[0468] Methods: Propionibacterium acnes was cultured in Brain Heart Infusion Agar (BHIA) for 48 hours under anaerobic conditions at 37°C. For MIC testing, 100 μl of BHI fermentation broth was added to all 96 wells, and 100 μl of drug-containing fermentation broth was added to the first well of each row (1A to 1H), and serial (double) dilutions were performed up to 10 wells (columns 1 to 10 of the 96-well plate). For bacterial inoculation, the turbidity of the Propionibacterium acnes culture was adjusted to 0.5 McFarland standard (approximately 1.5 × 10⁻⁶). 8 Cells / ml), and further diluted (100-fold diluted with sterile BHI fermentation broth). Except for the sterile control well (column 12 of the 96-well plate), the diluted Propionibacterium acnes suspension (100 μl) was added to each well.

[0469] For *Propionibacterium acnes*, the plates were incubated anaerobically at 37°C for 48 hours; for *Staphylococcus aureus*, the plates were incubated anaerobically at 37°C for 24 hours. The optical density at 595 nm was read using a Bio-Rad plate reader to generate dose-response curves. The minimum inhibitory concentrations (MICs) of the test compounds were recorded by adding Alamar blue dye.

[0470] Results: Table 6 and Figure 1C and Figure 1D The MIC and dose-response curves of different compounds in susceptible and resistant Propionibacterium acnes strains are shown. The results indicate that compound 91 exhibited a lower MIC value and a faster bactericidal curve in both bacterial strains (clindamycin-sensitive and resistant Propionibacterium acnes) compared to all other compounds. In contrast, compounds 115 and 116 (slightly different from compound 91) showed more than twice the MIC value obtained by compound 91 for both Propionibacterium acnes strains. Similarly, compounds 113 and 94 showed almost no activity against Propionibacterium acnes, suggesting the importance and involvement of free carbonyl and carboxyl groups in the superior activity of the compounds. Interestingly, inFigure 1C and Figure 1D In the dose-response curves, compound 90 was found to be inactive in clindamycin-sensitive Propionibacterium acnes, but showed promising activity in clindamycin-unresponsive Propionibacterium acnes strains.

[0471] In the presence of Staphylococcus aureus strains, all compounds 90, 91, 115 and 116 exhibited similar activities, but compound 113 showed minimal activity, which may be due to the spatial constraints involved in binding to the target protein (Table 6).

[0472] Conclusion: Compound 91 is a potent anti-acne agent and is effective against clindamycin-sensitive strains. It also showed better peak activity in clindamycin-resistant Propionibacterium acnes strains than in sensitive strains. This is also reflected in the dose-response curve. Figure 1C and Figure 1D In both strains, at very low concentrations, all Propionibacterium acnes bacteria were killed, indicating that it overcomes all mechanisms of antibiotic inresponsiveness, namely, resistance (described in the introduction) and resistance-related mechanisms. This suggests that the structure of compound 91 is unique, exhibiting higher biopotency against both susceptible and resistant Propionibacterium acnes strains than any other compound. The wide variation in structure-related molecules also underscores the fact that efficacy cannot be predicted solely based on structural similarity. Similarly, the higher MIC value of compound 91 against Staphylococcus aureus than against Propionibacterium acnes demonstrates the specificity of compound 91 against specific bacterial strains, and the results observed in one bacterial species cannot be arbitrarily transferred to another pathogenic bacterium.

[0473] Purity and bioactivity of compounds 90, 91, 94, 113, 115 and 116 (from Table 1A and Table 1B)

[0474] The purity of all the above-mentioned DART molecules was assessed by HPLC, and their bioactivity was evaluated in susceptible and resistant strains of Propionibacterium acnes and susceptible strains of Staphylococcus aureus. The results are shown in Table 6.

[0475] Table 6: HPLC purity of compounds and MIC values ​​against clindamycin-sensitive and clindamycin-resistant Propionibacterium acnes strains and laboratory Staphylococcus aureus strains.

[0476]

[0477] Example 12: Determination of DNA helicase activity using compounds 90, 91, 94, 113, 115 and 116 of Escherichia coli DNA helicase.

[0478] Materials: DNA gyrase assay kit (Topogen Inc.), proteinase K, chloroform, isoamyl alcohol, various test compounds, and agarose gel electrophoresis system.

[0479] Methods: DNA supercoiling activity was determined using a DNA gyrase assay kit (Topogen Inc.) containing relaxed pHOT1 E. coli plasmid DNA, according to the manufacturer's protocol. The standard reaction mixture (20 μl) contained 35 mM Tris-HCl (pH 7.5), 24 mM KCl, 4 mM MgCl2, 2 mM dithiothreitol, 1.8 mM spermine, 1 mM ATP, 6.5% glycerol, 0.1 mg / ml bovine serum albumin (BSA), 10 μg / ml relaxed pHOT1 plasmid DNA, and 1 U DNA gyrase. The reaction mixture was incubated at 37°C for 1 hour in the presence of different concentrations of the selected compound / fluoroquinolone. The reaction was terminated by adding 1 / 5 volume of the loading dye (4 μl), followed by proteinase K (50 μg / ml), and incubated again at 37°C for half an hour. 20 μl of chloroform / isoamyl alcohol (24:1) was added to each tube and briefly vortexed. The blue aqueous phase was then separated and analyzed by 1% agarose gel electrophoresis. The gel was stained with ethidium bromide for half an hour and then destained with water for 15 minutes. The gel was then visualized and photographed in a transilluminator.

[0480] Results: Compounds inhibited the supercoiling activity of DNA gyrase: Compound 91, with the best MIC value, was first selected to assess its effect on DNA gyrase activity. The intensity of the supercoiled band was observed at five different concentrations (range 0.25 μm to 5 μm) (Figure 2A). Compared to the untreated control (100%), the supercoiled band was reduced in the presence of compound 91 (approximately 54.3% at 0.25 μM, down to approximately 2% at 5 μM of compound 91) (…). Figure 2B ).

[0481] In addition, all compounds 90, 91, 94, 113, 115 and 116 were tested at two concentrations of 1 μM and 2.5 μM to examine their effects on DNA helicase activity. Figure 3A The results showed that all compounds inhibited the supercoiling activity of DNA gyrase. However, compounds 91 and 116 were highly potent in this regard compared to the other compounds, indicating that the minor structural differences between compounds 91 and 116 did not affect the binding affinity of the gyrase. However, compound 91 was observed to have a higher bactericidal efficacy than compound 116 in the dose-response curves. Figure 1C and Figure 1D This demonstrates that compound 91 may have a pattern that is quite different from other compounds.

[0482] When compound 91 was compared with the known fluoroquinolone naflufloxacin, it showed better inhibitory efficacy against DNA supercoiling by DNA gyrase at both tested concentrations. Figure 3B ).

[0483] Conclusion: The results obtained from the DNA gyrase activity assay indicate that compound 91 is the most potent in binding to and inhibiting DNA gyrase activity. Therefore, the mechanism of bacterial growth inhibition is through blocking DNA gyrase function, thereby preventing important cellular functions and ultimately leading to cell death. This compound demonstrates superior antibacterial potency and binding affinity compared to known fluoroquinolones and naflurane. These results are consistent with those obtained from MIC data and dose-response curves.

[0484] Example 13: Mutagenic concentration (MPC) of compound 91 compared to other known fluoroquinolones

[0485] Materials: Brain heart extract fermentation broth, Propionibacterium acnes MTCC 1951, petri dishes, autoclave, incubator, anaerobic chamber with anaerobic gas pack.

[0486] Methods: Propionibacterium acnes was cultured in brain heart infusion agar (BHIA) for 48 hours under anaerobic conditions at 37°C. Three to four culture dishes containing the 48-hour Propionibacterium acnes culture medium were then resuspended in sterile phosphate-buffered saline (PBS) (pH 7.2), and the turbidity was adjusted to a density of 1 (10⁻¹⁰) at 600 nm. 9 Cells / ml). Centrifuge 50 ml of this culture suspension at 4000 rpm for 40 minutes. Discard the supernatant and resuspend the precipitate in 250 μl of sterile PBS. Add 50 μl of this cell suspension (10 10 Cells were dispersed on plates containing various concentrations of antibiotics (around the MIC range). The plates were incubated at 37°C for 48 hours, and the lowest drug concentration at which no growth occurred was taken as its MPC. If a film was observed in plates with higher antibiotic concentrations, the film was further cultured on drug-free plates. The growth obtained on drug-free plates was then passaged on drug-containing plates (at the concentration at which film separation occurred). If no growth was observed on these plates at the end of the incubation period, the same concentration was confirmed as the MPC of the compound.

[0487] Results: Table 7 shows the MPC values ​​and MPC / MIC ratios of compound 91 against *Propionibacterium acnes*, compared to other known fluoroquinolones and clindamycin. The MPC / MIC ratios indicate that compound 91 is almost 3 times more active in preventing the development of resistance against *Propionibacterium acnes* than known anti-acne antibiotics such as naflufloxacin and the next-generation fluoroquinolone eulifloxacin, and twice as active as clindamycin. The MPC / MIC ratios of besifloxacin and compound 91 were found to be similar. This leads to the conclusion that both besifloxacin and compound 91 are effective molecules for the treatment and prevention of *Propionibacterium acnes*, and are ideal for preventing the development of resistance against the pathogen.

[0488] Conclusion: Unlike MIC testing (which typically uses approximately 10...) 4 -10 5 The inoculum volume is CFU / ml, and MPC calculation requires a large inoculum volume (approximately 10). 9 -10 10 (cfu / ml). Such a high inoculum was chosen to ensure the presence of a first-step resistance mutant within the susceptible bacterial community. Furthermore, compound 91 has an MPC / MIC of only 1.5 [SC1], while naloxone's MPC / MIC is almost 8. The narrower the window between the MIC and MPC of an antibiotic molecule, the less chance there is for selective mutant growth in the in vivo environment. This means that compound 91 will be more effective than other known anti-acne agents in preventing the development of resistance in the target microorganism.

[0489] Table 7: Mutagenic concentration (MPC) and MPC / MIC ratio of compound 91, and comparison with known fluoroquinolones and lincosamides.

[0490]

[0491] Example 14: Zone of Inhibition (ZOI) of the Topical Gel Formulation of Compound 91 compared to other commercially available formulations.

[0492] Materials: Brain heart extract fermentation broth, Staphylococcus aureus MTCC 6908, Propionibacterium acnes strains (MTCC1951 and CCARM 9010), 96-well plates, autoclave, incubator, anaerobic chamber with anaerobic gas pack, plate reader (595nm), Alamar blue.

[0493] Methods: For the ZOI test, 100 μl of bacterial suspension (0.5 McFarland standard equivalent) was spread onto a BHA plate. The test sample (formulation) was dissolved in water / solvent based on solubility. 10 μl of the test sample (with various concentrations of the compound) was loaded into a sterile tray (6 mm) and placed on a plate containing bacterial culture. For Propionibacterium acnes, the plate was incubated anaerobically at 37°C for 48 hours; for Staphylococcus aureus, the plate was incubated anaerobically at 37°C for 24 hours; their ZOI was then measured.

[0494] Results: The ZOI (measured in cm) for different test samples are shown below: Propionibacterium acnes 1951 (sensitive, Table 8); Propionibacterium acnes 9010 (resistant, Table 9); and Staphylococcus aureus (sensitive, Table 10). The formulation of compound 91 showed bactericidal curves against both bacterial strains, indicating that compound 91 retains its activity in the presence of other excipients present in the formulation. Interestingly, as in the DNA gyrase assay (… Figure 2A and Figure 3A )as well as Figure 1C and Figure 1D Supported by the dose-response curves visible in the images, ZOI data show that at low drug concentrations (1 μg–2 μg), compound 91 exhibits faster bactericidal activity against resistant Propionibacterium acnes compared to sensitive Propionibacterium acnes. Compound 91 is also effective against Staphylococcus aureus strains, and ZOI results support the MIC values ​​shown in Table 6.

[0495] Conclusion: The formulation of compound 91 exhibits activity against certain Gram-positive bacterial strains. In both clindamycin-sensitive and clindamycin-unresponsive Propionibacterium acnes, the formulation maintained the activity of compound 91 and was more effective in clindamycin-unresponsive Propionibacterium acnes, supporting the fact that the drug has specific biological activity.

[0496] Table 8-10: Zone of Inhibition (ZOI) of Compound 91 in a Topical Gel Formulation Compared to Clindamycin-Sensitive Propionibacterium acnes 1951 (A), Clindamycin-Unresponsive Propionibacterium acnes 9010 (B), and Laboratory Staphylococcus aureus Strain (B).

[0497] Table 8

[0498]

[0499] Table 9

[0500]

[0501] Table 10

[0502]

[0503] Example 15: Determination of the anti-inflammatory potential of compound 91 in THP-1 cells stimulated by Propionibacterium acnes

[0504] In this study, we selected compound 91 for anti-inflammatory assays because it exhibited a lower MIC, efficient gyrase binding, and a faster bacterial killing curve. The anti-inflammatory activity of compound 91 was investigated in THP-1 cells stimulated by Propionibacterium acnes (ATCC 6919).

[0505] Methods: Preparation of inflammatory stimulants: A suspension of Propionibacterium acnes culture was prepared in PBS, and the cell density was measured using a turbidimeter. The cell number in the suspension was adjusted to approximately 5 × 10⁻⁶ cells / mL. 8 CFU / ml. Then, the bacterial suspension was heat-inactivated at 80°C for 30 min and stored at -80°C until further use.

[0506] ELISA for studying inflammatory responses in THP-1 cells: Cells were seeded in a 96-well configuration (2 × 10⁶ cells per well). 5 THP-1 cells were seeded in medium containing 10% FBS. Cells were stimulated with 3 McFarland equivalents of heat-inactivated Propionibacterium acnes to induce inflammatory cytokines. Cells in control wells were treated with PBS. After one hour of induction with Propionibacterium acnes, the test reagent was added to the induced cells at the appropriate concentration (25 μg / ml for compound 91). The plate was incubated at 37°C for 24 hours. After 24 hours, the plate was centrifuged to precipitate the cells and the supernatant was collected. The cytokine levels of individual cytokines (IL-1α, IL-1β, IL-6, and IL-8) in the resulting cell culture supernatant were analyzed by ELISA using an R&D system kit, following the manufacturer's instructions.

[0507] Results: In Propionibacterium acnes-induced THP-1 cells, compound 91 exerted its anti-inflammatory effect by reducing IL-6: Heat-inactivated Propionibacterium acnes-induced THP-1 cells were treated with 25 μg / ml of compound 91, and the levels of IL-1α, IL-1β, IL-6, and IL-8 were subsequently analyzed in the culture supernatant. At the tested concentration, compound 91 significantly reduced the level of IL-6 induced by Propionibacterium acnes (nearly 60%). Figure 4A andTable 11). Using the known anti-inflammatory agent dexamethasone as a positive control, it showed a near 100% reduction in IL-6 levels. When treated with 25 μg / ml compound 91 (data not shown), THP-1 cells showed 90% viability. Compound 91 had no effect on IL-8 levels in Propionibacterium acnes-induced THP-1 cells. Figure 4B ). Figure 5A and Figure 5B The results shown indicate that compound 91 had little effect on IL-1α and IL-1β levels induced by Propionibacterium acnes (a reduction of approximately 20%–25%, as shown in Table 11). These results suggest that compound 91 is an effective anti-inflammatory agent in specific cases of inflammation induced by Propionibacterium acnes.

[0508] Conclusion: Results obtained from DNA gyrase activity assays (cell-free system) and anti-inflammatory assays in THP-1 cells indicate that compound 91 possesses a dual mode of action. In addition to inducing DNA damage from the nitrogen heterocyclic moiety, it also mediates one of the antibacterial mechanisms of action by targeting bacterial DNA gyrases, while its anti-inflammatory properties are manifested through its effects on inflammatory mediators in mammalian cells. In the case of acne, this dual mode of action may help reduce bacterial populations and inflammation at the lesion site, thereby providing faster healing and better patient compliance.

[0509] Table 11: Under the condition that the formation of cytokines induced by Propionibacterium acnes is 100%, compound 91 reduces the percentage of release of different cytokines IL-1α, IL-1β, IL-6 and IL-8 induced by Propionibacterium acnes in THP-1 cells.

[0510]

[0511] Example 16: Topical formulation of effective DART molecule 91

[0512] Cream and gel formulations of effective DART compound 91 are prepared, wherein the concentration of the active ingredient typically ranges from about 0.5 wt% to about 3 wt%, or from 0.5 wt% to 2 wt%, or most preferably from 1.0 wt% to 1.5 wt%. The formulations are maintained at a typical pH range of pH 4.0–8.0, or preferably pH 4.0–6.5, or most preferably pH 5.0–6.0. ​​The concentration of the active ingredient is sufficient to reduce, treat, or prevent skin infections and inflammation at the target tissue caused by Propionibacterium acnes, Staphylococcus aureus, or Staphylococcus epidermidis or other related anaerobic Gram-positive bacteria.

[0513] Based on solubility profiles of compound 91 in various solvents (e.g., dimethyl isosorbide, diethylene glycol monoethyl ether, PEG 400, propylene glycol, benzyl alcohol, and acetate buffer at pH 4.0), three different formulation strategies were employed to achieve improved pharmacokinetic / pharmacodynamic profiles, high skin penetration, and better drug deposition characteristics. These should result in faster reduction of bacterial counts and a more rapid decrease in host immune responses (e.g., inflammation). Such fast-acting, effective formulations will ultimately allow for reduced dosage and duration of treatment, thus ensuring better patient adherence. These formulations will not be limited to treating infections caused by Propionibacterium acnes (sensitive and resistant strains), but will also treat erythematous acne or atopic dermatitis or pustular dermatitis, or skin and skin structure infections or other bacterial skin infections caused by different families of Gram-positive anaerobic bacteria (e.g., Staphylococcus, Streptococcus, and other genera (sensitive and resistant strains)). More importantly, the formulation should be well-resistant to resistant bacteria and prevent any further development of resistance.

[0514] Composition Example 1: A topical formulation of a partially suspended API (compound 91) in gel form at a pH of 5.0-5.5 using hydroxyethyl cellulose (HEC) as a gelling agent (Table 12).

[0515] Table 12

[0516]

[0517] Preparation method:

[0518] 1. While maintaining a stirring speed of 100-150 rpm, add hydroxyethyl cellulose in batches to a measured volume of water, and allow the mixture to swell at 80-100 rpm for 1 hour (phase B).

[0519] 2. In another container, polyethylene glycol 400, propylene glycol and diethylene glycol monoethyl ether are mixed together, and compound 91 is added to the mixture in portions at 400 rpm for about 40-45 minutes to obtain a homogeneous dispersion (phase A).

[0520] 3. Slowly transfer the drug dispersion (phase A) to phase B and stir it at about 50-100 rpm for about 30 minutes to form a homogeneous mixture;

[0521] 4. Finally, add benzyl alcohol to the final mixture and mix for another 30 minutes at 50-100 rpm to obtain a white to light yellow gel formulation;

[0522] 5. Use a citric acid solution to maintain the final pH of the gel at 5.0 to 5.5.

[0523] Composition Example 2: Topical formulation of a partially suspended API (compound 91) in gel form at pH 5.0-5.5 using Carbopol 980 as a gelling agent (Table 13).

[0524] Table 13

[0525]

[0526] Preparation method:

[0527] 1. While stirring at 100-150 rpm, add the gelling agent Carbopol 980 in batches to the measured volume of water;

[0528] 2. The pH of the gel mixture was adjusted to 5.5 by adding triethanolamine solution, allowing Carbopol 980 to swell in water (phase B).

[0529] 3. In another container, polyethylene glycol, propylene glycol and diethylene glycol monoethyl ether are mixed together, and compound 91 is added in portions to the mixture while stirring at 400 rpm for about 40-45 minutes to obtain a homogeneous dispersion (phase A).

[0530] 4. Slowly transfer the drug dispersion (phase A) to phase B and stir it at about 50-100 rpm for about 30 minutes;

[0531] 5. Finally, add benzyl alcohol to the final mixture and stir at 50-100 rpm for another 30 minutes to obtain a white to pale yellow gel formulation;

[0532] 6. After preparing the gel, measure the pH and maintain the final pH between 5.0 and 5.5.

[0533] Composition Example 3: A topical formulation of a partially suspended API (compound 91) in gel form at pH 5.0-5.5, using propyl gallate as an antioxidant and EDTA as a buffer / chelating agent (Table 14).

[0534] Table 14

[0535]

[0536] Preparation method:

[0537] 1. While maintaining a stirring speed of 100-150 rpm, add hydroxyethyl cellulose in batches to a measured volume of water and allow it to swell at 80-100 rpm for 1 hour (phase B).

[0538] 2. In another container, polyethylene glycol 400, propylene glycol and diethylene glycol monoethyl ether are mixed together, and compound 91 is added to the mixture in portions at 400 rpm for about 40-45 minutes to obtain a homogeneous dispersion (phase A).

[0539] 3. Slowly add the drug dispersion (phase A) to phase B and stir at about 50-100 rpm for about 30 minutes;

[0540] 4. Finally, anhydrous ethylenediaminetetraacetic acid, benzyl alcohol and propyl gallate are added to the final mixture and stirred at 50-100 rpm for 30 minutes to obtain a white to light yellow gel formulation;

[0541] 5. Use citric acid solution to maintain the final pH of the prepared gel at 5.0 to 5.5.

[0542] Composition Example 4: Topical formulation of a fully suspended API (compound 91) in gel form at pH 5.0-5.5 using hydroxyethyl cellulose (HEC) as a gelling agent (Table 15).

[0543] Table 15

[0544]

[0545] Preparation method:

[0546] 1. While maintaining a stirring speed of 100-150 rpm, add hydroxyethyl cellulose in batches to a measured volume of water; allow the mixture to swell at 80-100 rpm for 1 hour (phase B).

[0547] 2. In another container, prepare an aqueous glycerol solution and add compound 91 to the mixture at 400 rpm to obtain a homogeneous dispersion (phase A).

[0548] 3. Slowly add the drug dispersion (phase A) to phase B and stir at about 50-100 rpm for about 30 minutes to obtain a homogeneous mixture;

[0549] 4. Finally, add propylparaben to the final mixture and mix for another 30 minutes at 50-100 rpm to obtain a white to pale yellow gel formulation;

[0550] 5. Use a citric acid solution to maintain the final pH of the gel at 5.0 to 5.5.

[0551] Composition Example 5: Topical formulation of a partially suspended API (compound 91) in cream form at pH 5.0-5.5 (Table 16)

[0552] Table 16

[0553]

[0554] Preparation method:

[0555] 1. While maintaining a stirring speed of 500 rpm, add hydroxyethyl cellulose in batches to a measured volume of water and heat at 50℃-55℃ (phase B).

[0556] 2. In another container, PEG-2 stearyl ether, PEG-20 stearyl ether, and cetearyl alcohol are heated at 50°C-55°C. Cyclopentasiloxane and isosorbide dimethyl ether are added to the mixture while stirring at 400 rpm. Compound 91 is added in portions to the final mixture while stirring at 400 rpm for about 5-10 minutes to obtain a homogeneous dispersion (phase A) at 50°C.

[0557] 3. Slowly add the drug dispersion (phase A) to phase B and stir at about 300-400 rpm for about 20-30 minutes until the temperature reaches 40°C;

[0558] 4. Finally, ethylenediaminetetraacetic acid dihydrate and benzyl alcohol are added to the final mixture and mixed at 400 rpm for about 30 minutes to obtain a white to light yellow cream formulation;

[0559] 5. Use a citric acid solution to maintain the final pH of the cream formulation at 5.0 to 5.5.

[0560] Composition Example 6: Topical formulation of a partially suspended API (compound 91) in cream form with a pH of 5.0-5.5 (Table 17)

[0561] Table 17

[0562]

[0563] Preparation method:

[0564] 1. While stirring at 100-150 rpm, add the gelling agent Carbopol 980 in batches to the measured volume of water;

[0565] 2. Adjust the pH of the gel mixture to 5.5 using a triethanolamine solution, allowing Carbopol 980 to swell in water, and then heat at 50°C-55°C (phase B).

[0566] 3. In another container, while maintaining a stirring speed of 400-500 rpm, add PEG-20 sorbitan monolaurate, sorbitan monolaurate, cetearyl alcohol, cyclopentasiloxane, and isosorbide dimethyl ether, and heat at 50-55°C. Add compound 91 in portions to the final mixture while stirring at 400 rpm for about 5-10 minutes to obtain a homogeneous dispersion (phase A) at 50°C.

[0567] 4. At 50℃-55℃ and with the stirring speed maintained at 400 rpm, slowly transfer the drug dispersion (phase A) to phase B, and cool it to 40℃ within 20-30 minutes;

[0568] 5. Finally, benzyl alcohol is added to the final mixture and cooled to room temperature to obtain a white to pale yellow cream formulation.

[0569] Example 17: Determination of the minimum inhibitory concentration (MIC) of different compounds and their formulations against Propionibacterium acnes strains using a microfermentation broth dilution method.

[0570] Materials: Brain heart extract fermentation broth, Propionibacterium acnes strains (MTCC and CCARM), 96-well plates, autoclave, incubator, anaerobic chamber, anaerobic gas pack, plate reader (600nm), Alamar blue.

[0571] Methods: Propionibacterium acnes (MTCC 3297, MTCC 1951, and CCARM 9010) were cultured in Brain Heart Infusion (BHI) fermentation broth for 48–72 hours under anaerobic conditions at 37°C. The test compounds / formulations were initially diluted with a suitable solvent and further diluted with BHI fermentation broth to obtain the desired concentrations. Samples of different concentrations (100 μl) (prepared by serial dilution) were added to 96-well plates. 100 μl of Propionibacterium acnes BHI fermentation broth culture was added to each well (the turbidity of the culture was adjusted to 0.5 McFarland standard (approximately 1.5 × 10⁻⁶)). 8 (and further diluted 100-fold with sterile BHI fermentation broth). In addition, growth control and sterile control were established using 100 μl of Propionibacterium acnes BHI fermentation broth culture and pure BHI fermentation broth, respectively.

[0572] The plates were incubated under anaerobic conditions at 37°C for 48–72 hours. The optical density at 595 nm was read using a Bio-Rad plate reader to generate dose-response curves. The minimum inhibitory concentrations (MICs) of the test compounds were recorded by adding Alamar blue dye.

[0573] Examples 18-22 and Tables 18-23 describe some exemplary novel formulations that contain a single API (e.g., besifloxacin) or an API in combination with adapalene.

[0574] Example 18: Micronized besifloxacin granular dispersion (D1)

[0575] Preparation: Besifloxacin was dispersed in a surfactant solution (2% aqueous solution of poloxamer 407). The resulting suspension was passed through a high-pressure homogenizer at approximately 800 bar. The output dispersion was collected in a beaker and circulated 10 times to obtain a dispersion of particles of appropriate size (particle size range of 2 μm to 8 μm). The size distribution was determined by MasterSizer (Malvern Instruments), and the average particle size was 4.1 μm [Dv(10)-0.8 μm, Dv(90)-8.9 μm].

[0576] Example 19: Preparation of gel / cream formulations containing besifloxacin alone and in combination with adapalene

[0577] Gel and / or cream formulations containing besifloxacin were formulated according to the compositions shown in Table 18. These gel formulations were off-white to pale yellow in appearance, with a pH of 5-5.5 and a viscosity of approximately 5000 mPa·s. The formulations contained besifloxacin equivalent to 1% (w / w) in three different forms: (1) micronized suspension of besifloxacin hydrochloride (Table 18, GL1, GL2); (2) fully soluble besifloxacin hydrochloride (Table 18, GL4); and (3) unsized besifloxacin particles suspended in a cream (CM1). In addition to besifloxacin formulations alone, besifloxacin was combined with adapalene (0.1%) (Table 18, GL1) to provide anti-acne and keratolytic activity for patients with acne.

[0578] Table 18: Gel and / or cream formulations of compositions GL1, GL2, GL3, GL4 and CM1

[0579]

[0580] Preparation method:

[0581] (1) Heat allantoin to 50°C until completely dissolved, then cool to room temperature;

[0582] (2) Add carboplatin to the above mixture and allow it to swell for 1-2 hours;

[0583] (3) Add the dispersion of micronized besifloxacin and adapalene powder to the swollen carboplatin mixture and stir it at 400 rpm for 30 minutes;

[0584] (4) Then, add glycerin, propylene glycol, PEG 400, poloxamer 407, followed by disodium EDTA dissolved in water, and then add phenoxyethanol to the above-stirred mixture. After adding all the ingredients, stir the mixture for 30 minutes.

[0585] (5) Neutralize the above mixture with triethanolamine and stir it at 800 rpm for 2-3 hours.

[0586] Example 20: Preparation of a cream formulation (CM1) containing besifloxaci...

Claims

1. Use of the formulation in the preparation of a medicament for the treatment or prevention of acne. In the formulation, besifloxacin is used in combination with another active agent, namely adapalene. in, The formulation is selected from one of GL1, S1, FW1, BW1, L1, SL1-SL3, GA5 and GL19 as shown in Tables 1-8: Table 1: Table 2: Table 3: Table 4: Table 5: Table 6: Table 7: Table 8: 。 2. The use as described in claim 1, wherein, The presence of Propionibacterium acnes, which causes the acne, but does not respond to therapeutic doses of clindamycin, minocycline, tetracycline, or erythromycin.

3. The use as described in claim 1, wherein, The intended use is to prepare a medicine for treating or preventing inflammation associated with acne.

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