6-CYCLOHEXYL-l-HYDROXY-4-METHYLPYRIDIN-2(lH)-ONE

AND DERIVATIVES THEREOF

INVENTORS

1. Mehmet Tanol

2. Scott J. Weir

CROSS-REFERENCE TO RELATED APPLICATIONS

[001] This patent application claims the benefit of U.S. Provisional Patent Application 61/419,218, filed on December 2, 2010, and which provisional application is incorporated herein by specific reference in their entirety.

BACKGROUND

[002] The molecule 6-cyclohexyl-l-hydroxy-4-methylpyridin-2(lH)-one, also known as Ciclopirox, is a commercially available topical antifungal agent. Ciclopirox has been used to treat superficial mycoses and Tinea versicolor. However, ciclopirox has a very poor solubility in aqueous fluids, which limits use in aqueous media and administration to a subject. As the human system is highly aqueous, ciclopirox is insufficient bioavailability due to the lack of water solubility. Often, ciclopirox is used in a salt form as ciclopirox olamine, but still has poor water solubility. As such, it would be beneficial to configure ciclopirox for improved water solubility in order to improve bioavailability.

BRIEF DESCRIPTION OF THE FIGURES

[003] The foregoing and following information as well as other features of this disclosure will become more fully apparent from the following description and appended claims, taken in conjunction with the accompanying drawings. Understanding that these drawings depict only several embodiments in accordance with the disclosure and are, therefore, not to be considered limiting of its scope, the disclosure will be described with additional specificity and detail through use of the accompanying drawings, in which:

[004] Figure 1 illustrates a schematic representation of a method of preparing 6- cyclohexyl-l-hydroxy-4-methylpyridin-2(lH)-one, which is referred to herein as Cyclopirox (Scheme 1).

[005] Figure 2 illustrates a schematic representation of a method of preparing di-tert- butyl(choloromethyl)phosphate (Scheme 2). [006] Figure 3 illustrates a schematic representation of a method of preparing a prodrug of 6-cyclohexyl-l-hydroxy-4-methylpyridin-2(lH)-one (Scheme 3).

[007] Figure 3A illustrates a schematic representation of a sub-reaction process identified as Scheme 3 A, which can be a sub-process of Scheme 3 of Figure 3.

[008] Figure 3B illustrates a mass spectroscopy chromatograph that shows the reaction products of Scheme 3A of Figure 3A.

[009] Figure 3C illustrates a mass spectrometry chromatograph that shows the presence of the free acid form of ((6-cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl dihydrogen phosphate.

[010] Figure 3D illustrates a dimer-type form of ciclopirox-POM prodrug.

[011] Figure 3E illustrates a mass spectrometry chromatograph of the dimer-type form of Ciclopirox-POM prodrug of Figure 3D.

[012] Figure 4 illustrates a schematic representation of salting ((6-cyclohexyl-4- methylpyridin-l(2H)-yl)oxy)methyl dihydrogen phosphate to obtain ((6-cyclohexyl-4- methylpyridin-l(2H)-yl)oxy)methyl phosphate disodium salt (Scheme 4).

[013] Figure 4A illustrates a mass spectrometry chromatograph that shows the presence of the ((6-cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl phosphate disodium salt of Figure 4.

[014] Figure 5 illustrates a schematic representation of a method of preparing dibenzyl(choloromethyl)phosphate (Scheme 5).

[015] Figure 6 illustrates a schematic representation of a method of preparing dibenzyl (((6-cyclohexyl-4-methyl-2-oxopyridin-l(2H)-yl)oxy)methyl) phosphate (Scheme 6).

[016] Figure 7 illustrates a schematic representation of a method of preparing ((6- cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl phosphate disodium salt (Scheme 7).

[017] Figure 8 illustrates a schematic representation of a method of preparing the POM reagent dibenzyl (choloromethyl)phosphate (Scheme 8).

DETAILED DESCRIPTION

[018] In the following detailed description, reference is made to the accompanying drawings, which form a part hereof. In the drawings, similar symbols typically identify similar components, unless context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described herein, and illustrated in the figures, can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are explicitly contemplated herein.

[019] Generally, the present invention relates to prodrugs of 6-cyclohexyl-l-hydroxy-4- methylpyridin-2(lH)-one, also known as ciclopirox. The ciclopirox prodrugs can include a phosphoryloxymethyl (POM) moiety in order to provide improve solubility and bioavailability to ciclopirox. The ciclopirox-POM of the present invention can also include derivatives of ciclopirox that include the POM prodrug moiety, which derivative prodrugs can be referred to herein generally under the term ciclopirox-POM or may be specifically referenced as ciclopirox-POM derivatives. The ciclopirox-POM prodrugs of the present invention can be prepared with traditional chemistry techniques, such as those described herein.

[020] The ciclopirox-POM prodrug has been shown to improve the aqueous solubility of ciclopirox, and thereby can improve its bioavailability. Accordingly, the ciclopirox-POM prodrug overcomes limitations with the formulation and/or delivery route of ciclopirox or other forms of ciclopirox, such as a ciclopirox salt like ciclopirox olamine. The improved solubility of the ciclopirox-POM prodrug can now allow for improved pharmaceutical compositions for administering effective amounts of ciclopirox. The pharmaceutical compositions can be formulated to be suitable for the route of administration, such as intravenous, oral, or transdermal.

[021] The ciclopirox-POM prodrug is configured so that the POM moiety is cleaved off by phosphatase enzymes in order to produce the original parent drug ciclopirox that is biologically active. As such, the ciclopirox-POM can be administered to a living subject, and then be enzymatically processed into bioactive ciclopirox within the body of the subject. While the subject usually will be human, the ciclopirox-POM prodrugs may be found to be suitable for a wide variety of animals, such as mammals, birds, reptiles, or the like.

[022] While ciclopirox has traditionally been used as a topical antifungal, the different formulations available for the ciclopirox-POM prodrugs of the present invention may provide therapeutically effective amounts of ciclopirox for other maladies that have been shown or that may be developed to be suitable for ciclopirox treatment. Accordingly, the ciclopirox-POM prodrugs may be used in therapies for: inhibition, treatment, and/or prevention of a fungus; inhibition, treatment, and/or prevention of cancer; inhibition, treatment, and/or prevention of dermatitis; inhibition, treatment, and/or prevention of superficial mycoses; for inhibition, treatment, and/or prevention of inflammation (e.g., as an anti-inflammatory, or NSAID); and inhibition, treatment, and/or prevention of one or more of tinea pedis, tinea cruris, and tinea corporis, Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton floccosum, and Microsporum canis, candidiasis (moniliasis), Candida albicans, tinea (pityriasis) vesicolor, or Malassezia furfur.

[023] The ciclopirox-POM prodrug improves bioavailability of ciclopirox such that increased amounts and distributions of the prodrug can be effective on cellular and subcellular levels. That is, more ciclopirox can be available throughout a subject as well as within individual cells of a subject. The increased amount of available ciclopirox improves the bioactivity and therapeutic potential, as well as improves the ability to modulate cellular processes. Accordingly, the ciclopirox-POM can be used for disrupting DNA repair, cell division, or intranuclear transport in a cell. The cells may be in vivo, ex vivo, or in vitro.

[024] While prodrugs of ciclopirox have been described, such prodrugs have been limited to conventional esters formed with available hydroxyl and/or amino groups, such as by acylation of activated acids in the presence of a base. The esters described can include produgs that have phenyl esters, aliphatic (C8-C24) esters, acyloxymethyl esters, carbamates, and amino acid esters (see WO 2010/048712, which is incorporated herein by specific reference in its entirety).

[025] Additionally, prodrugs with increased solubility have been created for compounds that include secondary and tertiary amine containing drugs. Examples of such prodrugs include N-phosphoryloxymethyl (POM) prodrugs where the methyl group of the POM moiety is coupled to a secondary or tertiary amine (see US 5,985,856, which is incorporated herein by specific reference in its entirety).

[026] It has now been found that alternative chemistry techniques can couple a POM prodrug entity to ciclopirox in order to form the ciclopirox-POM prodrugs of the present invention. The chemical synthesis protocols can conjugate ciclopirox, shown in below, through its hydroxyl group rather than through a secondary or tertiary amine. While ciclopirox does include a cyclic nitrogen, it has been found that such a nitrogen is undesirable to be linked to a POM due to the hydroxyl group or oxygen linked to the nitrogen possibly being important for the biological activity of ciclopirox. As such, the present invention provides chemistry techniques to conjugate a POM moiety to the nitrogen of ciclopirox.

[027] In one embodiment, the present invention includes a prodrug of ciclopirox or its derivatives. The prodrug can include a structure of Formula 1 or derivative thereof or stereoisomer thereof as well as a pharmaceutically acceptable salt thereof.

[028] In the formulae, R ^] -R ¹⁴ each independently can include a hydrogen, halogens, hydroxyls, alkoxys, straight aliphatics, branched aliphatics, cyclic aliphatics, substituted aliphatics, unsubstituted aliphatics, saturated aliphatics, unsaturated aliphatics, aromatics, polyaromatics, substituted aromatics, hetero-aromatics, amines, primary amines, secondary amines, tertiary amines, aliphatic amines, carbonyls, carboxyls, amides, esters, amino acids, peptides, polypeptides, derivatives thereof, substituted or unsubstituted, or combinations thereof as well as other well-known chemical substituents. R ¹³ and/or R ¹⁴ can be a positive ion such as a sodium ion, such that the compound forms a salt. Also, R ¹³ and/or R ¹⁴ independently can include a positive ion to form a salt with the prodrug or one or more of hydrogen, halogens, hydroxyls, alkoxys, straight aliphatics, branched aliphatics, cyclic aliphatics, substituted aliphatics, unsubstituted aliphatics, saturated aliphatics, unsaturated aliphatics, aromatics, polyaromatics, substituted aromatics, hetero- aromatics, amines, primary amines, secondary amines, tertiary amines, aliphatic amines, carbonyls, carboxyls, amides, esters, amino acids, peptides, polypeptides, or combinations thereof. The methyl linker can be expanded to a larger aliphatic group that is substituted or unsubstituted if desired, such that n can be about 0-20, more preferably 1-10, and most preferably 1-4. The linker may also have one or two oxygen atoms, such that m can be 0, 1 or 2.

[029] In one embodiment, R and/or R may independently be any one of various positive ions. In one example, R ¹³ and/or R ¹⁴ can independently include ions with positive charge of +1. The R ¹³ and/or R ¹⁴ can be an alkali metal. Also, the R ¹³ and/or R ¹⁴ can independently be a group 1 ion, such as a sodium ion. Accordingly, the bond between the oxygen and R ¹³ and/or R ¹⁴ can be covalent or ionic.

[030] In one embodiment, the R ¹³ and/or R ¹⁴ can independently be a protecting group. Examples of protecting groups can include tert-butyl and benzyl; however, other organic- based protecting groups can be used. These protecting groups may be left on the prodrug for administration, or removed prior to administration.

[031] In one embodiment, R ¹ is a short aliphatic, such as a methyl or other alkyl.

[032] In one embodiment, the R ² -R ¹² are all hydrogen.

[033] In one embodiment, the R ¹³ and/or R ¹⁴ are each independently a hydrogen.

[034] In one embodiment, the R ¹³ and/or R ¹⁴ are each independently a benzyl or tert- butyl.

[035] In one embodiment, the R ¹³ or R ¹⁴ includes a Ciclopirox-POM and the other is hydrogen, which is shown in Formula 10 below.

[036] In one embodiment, the present invention can include compounds with a structure of Formula 2 or derivative thereof or stereoisomer thereof. The n, m, R ¹ , and R ¹³ and/or 1 ⁴ can be the same as recited above for Formula 1.

[037] In one embodiment, the compounds can include the structure of Formula 3 derivative thereof or stereoisomer thereof. The n, m, R ¹ , and R ¹³ and/or R ¹⁴ can be same as recited above for Formula 1. Formula 3

[038] In one embodiment, the compounds can include the structure of Formula 4 or derivative thereof or stereoisomer thereof. The n and m can be the same as recited above

Formula 4

[039] In one embodiment, the compounds can include the structure of Formula 5 or derivative thereof or stereoisomer thereof. The n and m can be the same as recited above

5

[040] In one embodiment, the compounds can include the structure of Formula 6 or derivative thereof or stereoisomer thereof. The n and m can be the same as recited above for Formula 1. Formula 6

[041] In one embodiment, the compounds can include the structure of Formula 7 or derivative thereof or stereoisomer thereof. The n and m can be the same as recited above

Formula 7

[042] In one embodiment, the compounds can include the structure of Formula 8 or derivative thereof or stereoisomer thereof. The n and m can be the same as recited above

Formula 8

[043] In one embodiment, the compounds can include the structure of Formula 9 or derivative thereof or stereoisomer thereof. The n and m can be the same as recited above for Formula 1. Formula 9

[044] In one embodiment, the compounds can include the structure of Formula 10 or

Formula 10

[045] Any of the compounds may be prepared as pharmaceutically acceptable salts, if possible. Any common pharmaceutically acceptable salt ion can be used.

[046] Additionally, any of the compounds described herein and represented by the chemical formulae can have any of the R groups independently selected from substituents selected from the group of hydrogen, Ci -C ₂₄ alkyl, C ₂ -C ₂₄ alkenyl, C ₂ -C ₂₄ alkynyl, C5 - C ₂ o aryl, C ₆ -C ₂₄ alkaryl, C ₆ -C ₂₄ aralkyl, halo, hydroxyl, sulfhydryl, Ci -C ₂₄ alkoxy, C ₂ -C ₂₄ alkenyloxy, C ₂ -C ₂₄ alkynyloxy, C5 -C20 aryloxy, acyl (including C ₂ -C ₂₄ alkylcarbonyl (— CO-alkyl) and C ₆ -C ₂ o arylcarbonyl (— CO-aryl)), acyloxy (— O-acyl), C ₂ -C ₂₄ alkoxycarbonyl (— (CO)— O-alkyl), C ₆ -C ₂ o aryloxycarbonyl (— (CO)— O-aryl), halocarbonyl (— CO)— X where X is halo), C ₂ -C ₂₄ alkylcarbonato (— O— (CO)— O- alkyl), C ₆ -C ₂₀ arylcarbonato (— O— (CO)— O-aryl), carboxy (— COOH), carboxylato (— COO ^~ ), carbamoyl (— (CO)— NH ₂ ), mono-(Ci -C ₂₄ alkyl)-substituted carbamoyl (— (CO)— NH(Ci -C ₂₄ alkyl)), di-(Ci -C ₂₄ alkyl)-substituted carbamoyl (— (CO)— N(Ci -C ₂₄ alkyl) ₂ ), mono- substituted arylcarbamoyl (— (CO)— NH-aryl), thiocarbamoyl (— (CS)— NH ₂ ), carbamido (— NH— (CO)— NH ₂ ), cyano(— C≡N), isocyano (— N ⁺ ≡C ^" ), cyanato (— O— C≡N), isocyanato (— O— N ⁺ ≡C ^" ), isothiocyanato (— S— C≡N), azido (— N=N ⁺ =N ^" ), formyl (— (CO)— H), thioformyl (— (CS)— H), amino (— NH ₂ ), mono- and di-(Ci -C ₂₄ alkyl)-substituted amino, mono- and di-(Cs -C ₂ o aryl)-substituted amino, C ₂ -C ₂₄ alkylamido (— NH— (CO)-alkyl), C ₆ -C ₂₀ arylamido (— NH— (CO)-aryl), imino (— CR=NH where R is hydrogen, Ci -C ₂₄ alkyl, C5 -C20 aryl, C ₆ -C ₂₄ alkaryl, C ₆ -C ₂₄ aralkyl, etc.), alkylimino (— CR=N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (— CR=N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (— NO ₂ ), nitroso (—NO), sulfo (—SO ₂ —OH), sulfonate (— S ₂ — O ^" )' d -C ₂₄ alkylsulfanyl (— S- alkyl; also termed "alkylthio"), arylsulfanyl (— S-aryl; also termed "arylthio"), Ci -C ₂₄ alkylsulfinyl (— (SO)-alkyl), C ₅ -C ₂₀ arylsulfinyl (— (SO)-aryl), d -C ₂₄ alkylsulfonyl (— S0 ₂ -alkyl), C5 -C ₂ o arylsulfonyl (— S0 ₂ -aryl), phosphono (— P(0)(OH) ₂ ), phosphonato (— P(0)(0 ^" ) ₂ ), phosphinato (-P(0)(0-)), phospho (— P0 ₂ ), phosphino (— PH ₂ ), derivatives thereof, and combinations thereof.

PHARMACEUTICAL COMPOSITIONS

[047] In one embodiment, the ciclopirox-POM prodrug can be included in a pharmaceutical composition. The pharmaceutical compositions can be formulated to be suitable for the route of administration, such as intravenous, oral, or transdermal.

[048] In one embodiment, a pharmaceutical composition can include the ciclopirox- POM prodrug as described herein. For example, the pharmaceutical composition can include a pharmaceutically acceptable carrier. Since the ciclopirox-POM prodrug is now highly water soluble, the pharmaceutically acceptable can include water. However, the carrier can be sufficient in order to administer the ciclopirox-POM prodrug so that it reaches the aqueous environment of a subject to which it is administered, such as a human of the cells thereof.

[049] In one embodiment, the pharmaceutical composition can include the ciclopirox- POM prodrug being present in an amount greater than about 0.77%, more preferably greater than about 1%; or most preferably greater than about 2%.

[050] In one embodiment, the pharmaceutical composition can include water as a carrier.

[051] The compositions described herein can be prepared by per se known methods for the preparation of pharmaceutically acceptable compositions that can be administered to subjects, such that an effective quantity of the active substance is combined in a mixture with a pharmaceutically acceptable vehicle. Suitable vehicles are described, for example, in Remington's Pharmaceutical Sciences. On this basis, the compositions include, albeit not exclusively, solutions of the substances in association with one or more pharmaceutically acceptable vehicles or diluents, and contained in buffered solutions with a suitable pH and iso-osmotic with the physiological fluids. [052] In one embodiment, the effective amount of ciclopirox-POM prodrug is within the range of about 1 to about 200 mg/kg body weight of a subject. In one aspect, the effective amount of ciclopirox-POM prodrug is within the range of about 5 to about 50 mg/kg body weight. The ciclopirox-POM prodrug can be prepared into a solid dosage form that contains from about 20 mg to about 1000 mg of ciclopirox-POM prodrug. In one aspect, the composition can include about 20 mg to about 200 mg of ciclopirox-POM prodrug/kg body weight of subject, and can be formulated into a solid oral dosage form, a liquid dosage form, or an injectable dosage.

[053] In one embodiment, the present invention can include a pharmaceutical composition configured for treatment of a leukemic disorder. Such a composition can include an effective amount of ciclopirox-POM prodrug that can be administered orally or intravenous or other injection.

[054] Pharmaceutical compositions include, without limitation, lyophilized powders or aqueous or non-aqueous sterile injectable solutions or suspensions, which may further contain antioxidants, buffers, bacteriostats and solutes that render the compositions substantially compatible with the tissues or the blood of an intended recipient. Other components that may be present in such compositions include water, surfactants (e.g., Tween ^® ), alcohols, polyols, glycerin and vegetable oils, for example. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, tablets, or concentrated solutions or suspensions. The composition may be supplied, for example but not by way of limitation, as a lyophilized powder which is reconstituted with sterile water or saline prior to administration to the patient.

[055] Suitable pharmaceutically acceptable carriers include essentially chemically inert and nontoxic compositions that do not interfere with the effectiveness of the biological activity of the pharmaceutical composition. Examples of suitable pharmaceutical carriers include, but are not limited to, water, saline solutions, glycerol solutions, ethanol, N-(l (2,3- dioleyloxy)propyl)N,N,N-trimethylammonium chloride (DOTMA), diolesyl- phosphotidyl-ethanolamine (DOPE), and liposomes. Such compositions should contain a therapeutically effective amount of the compound, together with a suitable amount of carrier so as to provide the form for direct administration to the patient.

[056] The compositions described herein can be administered for example, by parenteral, intravenous, subcutaneous, intramuscular, intracranial, intraorbital, ophthalmic, intraventricular, intracapsular, intraspinal, intracisternal, intraperitoneal, intranasal, aerosol or oral administration. Common carriers or excipients can be used for preparing pharmaceutical compositions designed for such routes of administration.

[057] Another aspect provides a commercial package comprising a composition described herein, and associated therewith instructions for the use thereof for treatment of a disorder for which ciclopirox is effective, such as those described herein. For example, the package can include a suitable amount for treating a leukemic disorder, such as acute myeloid leukemia or acute lymphoid leukemia, in a subject in need of such treatment. In another embodiment. The commercial package can include instructions for administration and/or a therapeutic regimen for the treatment of chronic myelogenous leukemia, lymphoma or multiple myeloma. Another embodiment provides a commercial package comprising a composition described herein, and associated therewith instructions for the inducing cell death and/or inhibiting surviving activity or level in a leukemic disorder cell such as a leukemia cell.

[058] In one embodiment, the composition is devoid of an olamine. In fact, the prodrug can be devoid of an olamine. Accordingly, the composition can be devoid of a ciclopirox olamine or derivative thereof.

[059] In one embodiment, the prodrug can be present in a therapeutically effective amount for use as a therapy. For example, the therapeutically effective amount can be sufficient for one or more of the following: for use in treatment of a fungus; for use in treatment of cancer; for use in treatment of dermatitis; for disrupting DNA repair, cell division, or intranuclear transport in a cell; for use in treatment of superficial mycoses; for use as an anti-inflammatory; for use in treatment of one or more of tinea pedis, tinea cruris, and tinea corporis, Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton floccosum, and Microsporum canis, candidiasis (moniliasis), Candida albicans, tinea (pityriasis) vesicolor, or Malassezia furfur; for use in treating acute myeloid leukemia or acute lymphoid leukemia; chronic myelogenous leukemia, lymphoma or multiple myeloma, or others.

SYNTHESIS

[060] Generally, ciclopirox can be obtained and then reacted through the reaction protocols described herein in order to produce the ciclopirox-POM prodrug. Also, a ciclopirox derivative can be obtained and reacted following the synthetic protocols described herein, where a ciclopirox derivative reagent will result in a corresponding ciclopirox-POM derivative prodrug. In some instances, the ciclopirox or derivative thereof can be obtained in a salt, such as an olamine salt. Accordingly, the reaction scheme can include desalting the ciclopirox prior to conjugation with the POM prodrug moiety.

[061] In the figures and associated following descriptions of chemical reactions and the corresponding reactants, abbreviations are used to describe chemicals, which abbreviations are defined as follows: ED AC is N-(3-Dimethylaminopropyl)-N'- ethylcarbodiimide hydrochloride; DIAD is diisopropyl azodicarboxylate; BBDI is 1-tert- Butoxy-2-tert-butoxycarbonyl-l,2-dihydroisoquinoline; DICD is diisopropyl carbodiimide; and DBP is dibenzyl phosphate.

[062] In one embodiment, the present invention provides a method of preparing 6- cyclohexyl-l-hydroxy-4-methylpyridin-2(lH)-one, which method can include reaction Scheme 1 as shown in Figure 1. The reaction of Scheme 1 prepares a ciclopirox from a olamine salt thereof. Briefly, ciclopirox olamine (5 g, 18.6 mmol) can be dissolved in 2 N HC1, extracted with EtOAc, and precipitated with hexane in order to obtain ciclopirox (about an 84% yield).

[063] In one embodiment, the present invention provides a method of preparing a reagent for use in preparing a prodrug of 6-cyclohexyl-l-hydroxy-4-methylpyridin-2(lH)- one, which method can include reaction Scheme 2 as shown in Figure 2. The reaction Scheme 2 provides a reagent for preparation of a reagent of the POM prodrug moiety, which is di-tert-butyl(choloromethyl)phosphate. Briefly, potassium di-tert phosphate (5 g, 20 mmol) is dissolved in a minimum amount of cold water and 6 N HC1 is added drop- wise in order to form a precipitate, and then the precipitate is washed with cold water, which is then filtered and dried under vacuum in order to form di-tert phosphate. The di- tert-phosphate (4 g, 19 mmol) is then dissolved in about 100 mL acetone with tetramethylammonium hydroxide added drop-wise until reaching about pH 7, and then the solvent is removed and dried under vacuum to produce tetra-methyl ammonium di- tert-butyl phosphate. Tetra-methyl ammonium di-tert-butyl phosphate (5.11 g, 18 mmol) is then reacted with iodocholoro methane (CH ₂ CII) (25 g, 142 mmol, 7.88 equivalent) in about 150 ML DME and refluxed for about 2 hours before being filtered to remove the precipitate, removal of the solvent, and then dissolved in EA/H, and then filtered through a silica bed, and the solvent is removed and the product is dried to obtain di-tert- butyl(choloromethyl)phosphate. A TLC is shown to confirm product.

[064] In one embodiment, the present invention provides a method of preparing a prodrug of 6-cyclohexyl-l-hydroxy-4-methylpyridin-2(lH)-one (i.e., ((6-cyclohexyl-4- methylpyridin-l(2H)-yl)oxy)methyl phosphate disodium salt), which method can include reaction Scheme 3 as shown in Figure 3. The reaction Scheme 3 reacts ciclopirox with di-tert-butyl(choloromethyl)phosphate to form a ciclopirox-POM that includes a disodium salt. Briefly, ciclopirox (0.257 g, 1.25 mmol) and di-tert-butyl(choloromethyl)phosphate (0.450 g, 1.74 mmol) are dissolved in 5 mL and reacted with 60% NaH (0.62g, 154. mmol) at 0°C for 30 minutes and then room temperature for 2 hours before being quenched with water, and then the solvent is removed and process through a separation column with EA/H (1: 1) to yield di-tert-butyl (((6-cyclohexyl-4-methylpyridin-l(2H)- yl)oxy)methyl) phosphate, which was confirmed via TLC. The di-tert-butyl (((6- cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl) phosphate was then dissolved in 10 mL THF/CH2CI2 (3:1), and incubated at room temperature for 2 hours before the solvent was removed and purified via RPHPLC, and then equamolar Na2CC>3 in water/ACN was added and lyophilized to obtain ((6-cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl dihydrogen phosphate (14 mg, 0.044 mmol) and ((6-cyclohexyl-4-methylpyridin-l(2H)- yl)oxy)methyl phosphate disodium salt (15.89 mg), which is referred to herein as the primary ciclopirox-POM prodrug.

[065] Figure 3 A shows a sub-reaction process identified as Scheme 3 A, which can be a sub-process of Scheme 3. Scheme 3 A shows the di-tert-butyl (((6-cyclohexyl-4- methylpyridin-l(2H)-yl)oxy)methyl) phosphate converting to tert-butyl (((6-cyclohexyl- 4-methylpyridin-l(2H)-yl)oxy)methyl) hydrogen phosphate, which converts to ((6- cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl dihydrogen phosphate. Figure 3B shows a mass spectrometry chromatograph that shows the presence of these chemical entities.

[066] The free acid form of the ciclopirox-POM prodrug (i.e., ((6-cyclohexyl-4- methylpyridin-l(2H)-yl)oxy)methyl dihydrogen phosphate) was also purified by RPHPLC as shown in Figure 3C. The purified product was shown to have a mass spectrometry chromatograph as shown in Figure 3C.

[067] Additionally, another dimer-type form of ciclopirox-POM prodrug has been created as shown in Figure 3D, which is referred to as the ciclopirox-POM dimer prodrug. The ciclopirox-POM dimer prodrug is confirmed with mass spectrometry chromatograph as shown in Figure 3E.

[068] Figure 4 illustrates Scheme 4 which is the process of salting ((6-cyclohexyl-4- methylpyridin-l(2H)-yl)oxy)methyl dihydrogen phosphate with sodium ions in order to obtain ((6-cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl phosphate disodium salt, which is confirmed with mass spectrometry chromatograph as shown in Figure 4A.

[069] In one embodiment, the present invention can include a method of preparing the POM reagent dibenzyl(choloromethyl)phosphate, as shown in reaction Scheme 5 of Figure 5. Briefly, CH ₂ CII (5 mL; 68.6 mmol) is reacted with silver dibenzyl phosphate (2 g, 52 mmol) in about 25 mL toluene and refluxed for about 1 hour before the solvent is removed and processed through a separation column with 1/lEA/H in order to produce dibenzyl(choloromethyl)phosphate as confirmed via TLC.

[070] In one embodiment, the present invention provides a method of preparing a prodrug dibenzyl (((6-cyclohexyl-4-methyl-2-oxopyridin- l(2H)-yl)oxy)methyl) phosphate, which is shown by Scheme 6 of Figure 6. Briefly, ciclopirox (0.431 g, 2.08 mmol) is dissolved in 10 mL DMF with dibenzyl(choloromethyl)phosphate (0.920 g, 2.81 mmol) along with 60% NaH (0.096 g, 2.38 mmol) and incubated at 0°C for 30 minutes and then at room temperature for 1 hour before being quenched with water. The solvent is then removed and processed through a separation column with EA/H 1 : 1 in order to yield dibenzyl (((6-cyclohexyl-4-methyl-2-oxopyridin-l(2H)-yl)oxy)methyl) phosphate, which is confirmed via the TLC as shown.

[071] In one embodiment, dibenzyl (((6-cyclohexyl-4-methyl-2-oxopyridin-l(2H)- yl)oxy)methyl) phosphate is converted into ((6-cyclohexyl-4-methylpyridin-l(2H)- yl)oxy)methyl phosphate disodium salt as shown in Scheme 7 of Figure 7. Briefly, dibenzyl (((6-cyclohexyl-4-methyl-2-oxopyridin-l(2H)-yl)oxy)methyl) phosphate is dissolved in 25 mL THF in the presence of 100 mg Pd/C, and under hydrogen (H ₂ ) at room temperature for 3 hours before the solvent was removed and the product dissolved in ACN. Na ₂ C0 ₃ in water is then added and lyophilized. The product is determined to be a minor amount of ((6-cyclohexyl-4-methylpyridin-l(2H)-yl)oxy)methyl dihydrogen phosphate (about 0.273 g,0.86 mmol) and the major product being ((6-cyclohexyl-4- methylpyridin-l(2H)-yl)oxy)methyl phosphate disodium salt (0.310 g), which was confirmed via TLC as shown.

[072] In one embodiment, another method of preparing the POM reagent dibenzyl(choloromethyl)phosphate, as shown in reaction Scheme 8 of Figure 8. Briefly, dibenzylphosphate (10.6 g; 38.08 mmol) and chloromethyl sulfochloridate (7.52g, 456.6 mmol/120 mL CH ₂ C1 ₂ ) are dissolved in 320 mL water and 200 mL CH ₂ C1 ₂ with NaHC0 ₃ (12.80 g, 152.32 mmol) and n-Bu ₄ NHS0 ₄ (12.94 g, 38.08 mmol) at 0°C to room temperature overnight, and then processed through a separation column with ½ EA/H, EA. This reaction yields about 48% of dibenzyl(choloromethyl)phosphate (6 g), which is confirmed with TLC as shown.

[073] In one embodiment, the present invention provides another method of preparing a prodrug dibenzyl (((6-cyclohexyl-4-methyl-2-oxopyridin- l(2H)-yl)oxy)methyl) phosphate, which is shown by Scheme 6 of Figure 6. Briefly, Ciclopirox (1 g, 4.82 mmol) is dissolved in 20 mL DMF with dibenzyl(choloromethyl)phosphate (2 g, 6.12 mmol) along with 60% NaH (0.375 g) and incubated at 0°C for 30 minutes and then at room temperature for 1 hour before being quenched with water. The solvent is then removed, and the product is dissolved in EtOAc, washed with water, solvent is removed, and processed through a separation column with EA/H 1:2, and RPHPLC in order to yield 67% dibenzyl (((6-cyclohexyl-4-methyl-2-oxopyridin-l(2H)-yl)oxy)methyl) phosphate (1.616 g).

EXPERIMENTAL

[074] The ciclopirox-POM prodrug described herein (((6-cyclohexyl-4-methylpyridin- l(2H)-yl)oxy)methyl phosphate disodium salt, MW 361) was studied and compared with the ciclopirox free base (MW 207) and ciclopirox olamine (MW 268). Equivalent doses of ciclopirox for these three entities were administered such as follows: ciclopirox olamine at 2.0 mg/mL and ciclopirox-POM prodrug at 2.69 mg/mL. The plasma ciclopirox concentrations in (ng/mL) were determined following bolus administration of ciclopirox-POM prodrug at a 10 mg/kg IV bolus in a mouse model. Time points where data collected are shown in Table 1 to be at 5, 15, 30, 60, 90, 120, 180, and 240 minutes along with the data. Additional Samples were collected were collected at 360, 480, 720 and 1440 minutes, however, concentrations fell below BQL of 10 ng/mL, and are not included in PK data analysis. The IV bolus formulation for prodrug was in 25 mM phosphate buffer.

Table 1 - Plasma Ciclopirox Concentrations (ng/mL): Ciclopirox-POM IV Bolus

Time

A B C D E F Mean SD %CV (min)

5 7280 7320 6370 9700 8500 8860 8005 1226.1 15.3

15 3310 3180 1520 1370 4010 3190 2763.3 1067.3 38.6

30 994 1200 986 1030 1080 452 957 259.5 27.1

60 410 259 284 259 409 436 342.8 83.8 24.4

90 97.6 157 74.2 79.1 162 168 123 43.9 35.7

120 58 73.7 57.9 19.3 71.7 84.8 60.9 22.8 37.4

180 18.1 23 19.5 34.8 19.7 37.7 25.5 8.6 33.6

240 13 10 15 12 10 8 11.1 2.6 23.6

[075] The plasma ciclopirox concentrations in (ng/mL) were determined following bolus administration of ciclopirox olamine at a 10 mg/kg IV bolus in a mouse model. Time points where data collected are shown in Table 1 to be at 5, 15, 30, 60, 90, 120, 180, and 240 minutes along with the data. The IV bolus formulation for ciclopirox olamine was in 0.05 M Captisol® and 25 mM phosphate buffer, and the corresponding data is shown below in Table 2. Additional Samples were collected were collected at 360, 480, 720 and 1440 minutes, however, concentrations fell below BQL of 10 ng/mL, and are not included in PK data analysis

Table 2 - Plasma Ciclopirox Concentrations (ng/mL): Ciclopirox Olamine IV Bolus

Time

A B C D E F Mean SD %CV (min)

5 6090 3490 5830 4480 3930 5430 4875 1064.3 21.8

15 543 591 1410 1100 739 968 891.8 332.6 37.3

30 545 371 166 339 336 364 353.5 120.6 34.1

60 171 127 124 144 110 124 133.3 21.4 16.1

90 45.2 63.4 58.8 42.9 58 84 58.7 14.8 25.2

120 46.1 41.2 46.9 39.2 42.4 OL 43.2 17.6 48.9

180 14 18.9 18.1 28.7 47.2 24.5 25.2 11.9 47.3

240 12 12 21 16 OL 19 15.9 4.2 26.6

[076] The plasma ciclopirox (i.e., CPX) concentrations in (ng/mL) were determined following bolus administration of ciclopirox-POM prodrug at a 30 mg/kg orally in a mouse model. Time points where data collected are shown in Table 3 to be at 5, 15, 30, 60, 90, 120, 180, and 240 minutes along with the data. Additional Samples were collected were collected at 360, 480, 720 and 1440 minutes, however, concentrations fell below BQL of 10 ng/mL, and are not included in PK data analysis. The oral formulation is a composition having Orasweet SF along with the ciclopirox- PIM prodrug.

Table 3 - Plasma Ciclopirox Concentrations (ng/mL): Ciclopirox-POM Oral Dose

Time

A B C D E F Mean SD %CV (min)

0 0 0 0 0 0 0 0 ND ND

15 388 646 857 245 479 507 520.3 211.8 40.7

30 302 252 264 317 207 342 280.7 49.1 17.5

60 129 404 154 229 171 207 215.7 99 45.9

90 91 66.8 146 137 120 80.3 106.9 32.2 30.1

120 72.5 60.6 64.6 81.4 83.6 OL 72.5 39.6 54.6

180 36.8 45.1 59.1 66.4 44.9 44 49.4 11 22.4

240 9.51 OL 19 14.7 38.1 19 20.1 10.8 53.9

[077] The plasma ciclopirox concentrations in (ng/mL) were determined following bolus administration of ciclopirox olamine at a 30 mg/kg oral dose in a mouse model. Time points where data collected are shown in Table 1 to be at 5, 15, 30, 60, 90, 120, 180, and 240 minutes along with the data. The oral formulation for ciclopirox olamine was a suspension of Orasweet SF, and the corresponding data is shown below in Table 4. Additional Samples were collected were collected at 360, 480, 720 and 1440 minutes, however, concentrations fell below BQL of 10 ng/mL, and are not included in PK data analysis

Table 4 - Plasma Ciclopirox Concentrations (ng/mL): Ciclopirox Olamine Oral Dose

Time

A B C D E F Mean SD %CV (min)

0 0 0 0 0 0 0 0 ND ND

15 478 OL 625 445 565 626 547.8 83.4 15.2

30 427 376 430 432 388 641 449 97 21.6

60 379 319 501 211 246 196 308.7 116.8 37.8

90 124 297 259 348 337 198 260.5 86.5 33.2

120 OL 101 146 85.4 204 218 150.9 59.4 39.4

180 78.2 83.4 76.4 107 66.5 69.2 80.1 14.5 18.1

240 72.3 93.1 92.5 48.9 67.8 73.6 74.7 16.6 22.2

[078] Additionally, mouse intravenous dose pharmacokinetics were determined. Briefly, the pharmacokinetics were determined for mouse hepatic blood flow at 90 mL/min/kg, an ciclopirox would be considered a medium clearance drug with extraction ration of 0.6 The IV bolus pharmacokinetics are shown in Table 5 below.

Table 5 - Ciclopirox Pharmacokinetics (10 mg/kg IV Bolus)

PK Parameter CPX Prodrug CPX Olamine

10 mg/kg 10 mg/kg

Alpha rate constant (min-1) 0.07043 0.08802

Alpha Half-life (min) 9.84 7.87

Beta rate constant (min-1) 0.01929 0.01208

Beta Half-life (min) 35.92 57.35

AUC (ng/mL/kg) 160330 140313

CL (mL/min/kg) 48.275* 55.162*

Vdbeta (mL/kg) 2502 4565.2

[079] Additionally, mouse oral dose pharmacokinetics were determined. Briefly, the pharmacokinetics were determined for mouse hepatic blood flow at 90 mL/min/kg, an ciclopirox would be considered a medium clearance drug with extraction ration of 0.6 The oral pharmacokinetics are shown in Table 6 below.

Table 6 - Ciclopirox Pharmacokinetics (30 mg/kg Oral)

CPX Prodrug CPX OIamine

PK Parameter

30 mg/kg 30 mg/kg

Cmax (ng/mL) 520.3 547.8

Tmax (min) 15 15

Beta rate constant (min-1) 0.01316 0.00916

Beta Half-life (min) 52.76 75.67

AUC (ng/mL/kg) 32157.91 57389.8

CL/F (mL/min/kg) 722.06 404.602

[080] The IV bolus and oral administration of ciclopirox-POM prodrug and ciclopirox olamine were also studied with regard to bioavailability of ciclopirox. The data is shown in Table 7. The IV bolus and oral administrations are described above.

Table 7 - Bioavailability of Ciclopirox (IV Bolus v. Oral) m PK, n Parame ,ter ^{IV 0ra l}

Administration Administration

CPX AUC (ng/mL/kg)

Prodrug 160330.3 32157.91

Olamine Salt 140312.6 57389.8

Foral

Prodrug 0.066858

Olamine Salt 0.136338

CPX Bioavailability

Following Prodrug

Administration 1.142665 0.560342 [081] The data in the foregoing tables indicates that ciclopirox-POM prodrug as described herein can be an improvement for administration to subjects. Based on intravenous pharmacokinetics following administration of ciclopirox olamine, ciclopirox is considered a medium clearance drug based on intravenous clearance compared to hepatic blood flow in the mouse. It was found that ciclopirox was completely bioavailable when given IV as the prodrug, compared to IV administration of ciclopirox olamine, and thereby the ciclopirox-POM is an improvement in chemical formulation. The data shows that ciclopirox was 50% bioavailable, when given as orally as the prodrug, compared to CPX Olamine administered as an oral suspension, which shows an improvement in ciclopirox-POM prodrug. Accordingly, ciclopirox is readily bioavailable when given IV as the prodrug. Based on the inability to detect the prodrug in plasma following IV and oral administration, it appears the prodrug is rapidly metabolized to ciclopirox when it reaches the systemic circulation. Thus, ciclopirox-POM prodrug has advantages over ciclopirox olamine, from a physicochemical properties standpoint, and can be formulated into a suitable IV product. Based on the data, it is reasonable to believe that the ciclopirox-POM prodrug is rapidly converted back to ciclopirox.

[082] The term "alkyl" or "aliphatic" as used herein refers to a branched or unbranched saturated hydrocarbon group typically although not necessarily containing 1 to about 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, and the like, as well as cycloalkyl groups such as cyclopentyl, cyclohexyl, and the like. Generally, although again not necessarily, alkyl groups herein contain 1 to about 18 carbon atoms, or 1 to about 12 carbon atoms. The term "lower alkyl" intends an alkyl group of 1 to 6 carbon atoms. Substituents identified as "Ci -C ₆ alkyl" or "lower alkyl" contains 1 to 3 carbon atoms, and such substituents contain 1 or 2 carbon atoms (i.e., methyl and ethyl). "Substituted alkyl" refers to alkyl substituted with one or more substituent groups, and the terms "heteroatom-containing alkyl" and "heteroalkyl" refer to alkyl in which at least one carbon atom is replaced with a heteroatom, as described in further detail infra. If not otherwise indicated, the terms "alkyl" and "lower alkyl" include linear, branched, cyclic, unsubstituted, substituted, and/or heteroatom-containing alkyl or lower alkyl, respectively.

[083] The terms "alkenyl" as used herein refers to a linear, branched or cyclic hydrocarbon group of 2 to about 24 carbon atoms containing at least one double bond, such as ethenyl, n-propenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, tetracosenyl, and the like. Generally, although again not necessarily, alkenyl groups herein contain 2 to about 18 carbon atoms, or 2 to 12 carbon atoms. The term "lower alkenyl" intends an alkenyl group of 2 to 6 carbon atoms, and the specific term "cycloalkenyl" intends a cyclic alkenyl group, or having 5 to 8 carbon atoms. The term "substituted alkenyl" refers to alkenyl substituted with one or more substituent groups, and the terms "heteroatom-containing alkenyl" and "heteroalkenyl" refer to alkenyl in which at least one carbon atom is replaced with a heteroatom. If not otherwise indicated, the terms "alkenyl" and "lower alkenyl" include linear, branched, cyclic, unsubstituted, substituted, and/or heteroatom-containing alkenyl and lower alkenyl, respectively.

[084] The term "alkynyl" as used herein refers to a linear or branched hydrocarbon group of 2 to 24 carbon atoms containing at least one triple bond, such as ethynyl, n- propynyl, and the like. Generally, although again not necessarily, alkynyl groups herein contain 2 to about 18 carbon atoms, or 2 to 12 carbon atoms. The term "lower alkynyl" intends an alkynyl group of 2 to 6 carbon atoms. The term "substituted alkynyl" refers to alkynyl substituted with one or more substituent groups, and the terms "heteroatom- containing alkynyl" and "heteroalkynyl" refer to alkynyl in which at least one carbon atom is replaced with a heteroatom. If not otherwise indicated, the terms "alkynyl" and "lower alkynyl" include linear, branched, unsubstituted, substituted, and/or heteroatom- containing alkynyl and lower alkynyl, respectively.

[085] The term "alkoxy" as used herein intends an alkyl group bound through a single, terminal ether linkage; that is, an "alkoxy" group may be represented as— O-alkyl where alkyl is as defined above. A "lower alkoxy" group intends an alkoxy group containing 1 to 6 carbon atoms, and includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, t- butyloxy, etc. Substituents identified as "Ci -C ₆ alkoxy" or "lower alkoxy" herein contain 1 to 3 carbon atoms, and such substituents contain 1 or 2 carbon atoms (i.e., methoxy and ethoxy).

[086] The term "aryl" as used herein, and unless otherwise specified, refers to an aromatic substituent containing a single aromatic ring or multiple aromatic rings that are fused together, directly linked, or indirectly linked (such that the different aromatic rings are bound to a common group such as a methylene or ethylene moiety). Examples of aryl groups contain 5 to 20 carbon atoms, and aryl groups contain 5 to 14 carbon atoms. Exemplary aryl groups contain one aromatic ring or two fused or linked aromatic rings, e.g., phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, benzophenone, and the like. "Substituted aryl" refers to an aryl moiety substituted with one or more substituent groups, and the terms "heteroatom-containing aryl" and "heteroaryl" refer to aryl substituent, in which at least one carbon atom is replaced with a heteroatom, as will be described in further detail infra. If not otherwise indicated, the term "aryl" includes unsubstituted, substituted, and/or heteroatom-containing aromatic substituents.

[087] The term "aryloxy" as used herein refers to an aryl group bound through a single, terminal ether linkage, wherein "aryl" is as defined above. An "aryloxy" group may be represented as— O-aryl where aryl is as defined above. Examples of aryloxy groups contain 5 to 20 carbon atoms, and aryloxy groups contain 5 to 14 carbon atoms. Examples of aryloxy groups include, without limitation, phenoxy, o-halo-phenoxy, m-halo- phenoxy, p-halo-phenoxy, o-methoxy-phenoxy, m-methoxy-phenoxy, p-methoxy- phenoxy, 2,4-dimethoxy-phenoxy, 3,4,5-trimethoxy-phenoxy, and the like.

[088] The term "alkaryl" refers to an aryl group with an alkyl substituent, and the term "aralkyl" refers to an alkyl group with an aryl substituent, wherein "aryl" and "alkyl" are as defined above. Examples of aralkyl groups contain 6 to 24 carbon atoms, and aralkyl groups contain 6 to 16 carbon atoms. Examples of aralkyl groups include, without limitation, benzyl, 2-phenyl-ethyl, 3 -phenyl-propyl, 4-phenyl-butyl, 5-phenyl-pentyl, 4- phenylcyclohexyl, 4-benzylcyclohexyl, 4-phenylcyclohexylmethyl, 4- benzylcyclohexylmethyl, and the like. Alkaryl groups include, for example, p- methylphenyl, 2,4-dimethylphenyl, p-cyclohexylphenyl, 2,7-dimethyinaphthyl, 7- cyclooctylnaphthyl, 3-ethyl-cyclopenta-l,4-diene, and the like.

[089] The term "cyclic" refers to alicyclic or aromatic substituents that may or may not be substituted and/or heteroatom containing, and that may be monocyclic, bicyclic, or polycyclic.

[090] The terms "halo" and "halogen" are used in the conventional sense to refer to a chloro, bromo, and fluoro or iodo substituent.

[091] The term "heteroatom-containing" as in a "heteroatom-containing alkyl group" (also termed a "heteroalkyl" group) or a "heteroatom-containing aryl group" (also termed a "heteroaryl" group) refers to a molecule, linkage or substituent in which one or more carbon atoms are replaced with an atom other than carbon, e.g., nitrogen, oxygen, sulfur, phosphorus or silicon, typically nitrogen, oxygen or sulfur. Similarly, the term "heteroalkyl" refers to an alkyl substituent that is heteroatom-containing, the term "heterocyclic" refers to a cyclic substituent that is heteroatom-containing, the terms "heteroaryl" and heteroaromatic" respectively refer to "aryl" and "aromatic" substituents that are heteroatom-containing, and the like. Examples of heteroalkyl groups include alkoxyaryl, alkylsulfanyl-substituted alkyl, N-alkylated amino alkyl, and the like. Examples of heteroaryl substituents include pyrrolyl, pyrrolidinyl, pyridinyl, quinolinyl, indolyl, pyrimidinyl, imidazolyl, 1 ,2,4-triazolyl, tetrazolyl, etc., and examples of heteroatom-containing alicyclic groups are pyrrolidino, morpholino, piperazino, piperidino, etc.

[092] The term "hydrocarbyl" refers to univalent hydrocarbyl radicals containing 1 to about 30 carbon atoms, or 1 to about 24 carbon atoms, or 1 to about 18 carbon atoms, or about 1 to 12 carbon atoms, including linear, branched, cyclic, saturated, and unsaturated species, such as alkyl groups, alkenyl groups, aryl groups, and the like. "Substituted hydrocarbyl" refers to hydrocarbyl substituted with one or more substituent groups, and the term "heteroatom-containing hydrocarbyl" refers to hydrocarbyl in which at least one carbon atom is replaced with a heteroatom. Unless otherwise indicated, the term "hydrocarbyl" is to be interpreted as including substituted and/or heteroatom-containing hydrocarbyl moieties.

[093] By "substituted" as in "substituted alkyl," "substituted aryl," and the like, as alluded to in some of the aforementioned definitions, is meant that in the alkyl, aryl, or other moiety, at least one hydrogen atom bound to a carbon (or other) atom is replaced with one or more non-hydrogen substituents.

[094] In addition, the aforementioned functional groups may, if a particular group permits, be further substituted with one or more additional functional groups or with one or more hydrocarbyl moieties such as those specifically enumerated above. Analogously, the above-mentioned hydrocarbyl moieties may be further substituted with one or more functional groups or additional hydrocarbyl moieties such as those specifically enumerated.

[095] When the term "substituted" appears prior to a list of possible substituted groups, it is intended that the term apply to every member of that group. For example, the phrase "substituted alkyl, alkenyl, and aryl" is to be interpreted as "substituted alkyl, substituted alkenyl, and substituted aryl." Analogously, when the term "heteroatom-containing" appears prior to a list of possible heteroatom-containing groups, it is intended that the term apply to every member of that group. For example, the phrase "heteroatom- containing alkyl, alkenyl, and aryl" is to be interpreted as "heteroatom-containing alkyl, heteroatom-containing alkenyl, and heteroatom-containing aryl."

[096] One skilled in the art will appreciate that, for this and other processes and methods disclosed herein, the functions performed in the processes and methods may be implemented in differing order. Furthermore, the outlined steps and operations are only provided as examples, and some of the steps and operations may be optional, combined into fewer steps and operations, or expanded into additional steps and operations without detracting from the essence of the disclosed embodiments.

[097] The present disclosure is not to be limited in terms of the particular embodiments described in this application, which are intended as illustrations of various aspects. Many modifications and variations can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the disclosure, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present disclosure is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that this disclosure is not limited to particular methods, reagents, compounds compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[098] With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity.

[099] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "includes" should be interpreted as "includes but is not limited to," etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles "a" or "an" limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases "one or more" or "at least one" and indefinite articles such as "a" or "an" (e.g., "a" and/or "an" should be interpreted to mean "at least one" or "one or more"); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of "two recitations," without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to "at least one of A, B, and C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., " a system having at least one of A, B, and C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). In those instances where a convention analogous to "at least one of A, B, or C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., " a system having at least one of A, B, or C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase "A or B" will be understood to include the possibilities of "A" or "B" or "A and B."

[0100] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0101] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as "up to," "at least," and the like include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.

[0102] From the foregoing, it will be appreciated that various embodiments of the present disclosure have been described herein for purposes of illustration, and that various modifications may be made without departing from the scope and spirit of the present disclosure. Accordingly, the various embodiments disclosed herein are not intended to be limiting, with the true scope and spirit being indicated by the following claims. All references recited or in the incorporated references herein are incorporated herein by specific reference in their entirety.