COMPOUNDS AND METHODS FOR THE TREATMENT OF DERMAL AND OCULAR DISORDERS

Abstract

Described herein are compositions and methods for the treatment or prevention of dermal or ocular surface disorders. Ocular surface disorders include ocular allergy, dry eye disease, ocular manifestation of graft versus host disease and other inflammatory and/or infectious diseases of the (e.g., anterior) surface of the eye(s). Said compositions and methods comprise keratolytic conjugates which demonstrate immunological, keratolytic, anti-inflammatory, and/or other desirable activities. Topical administration of said compositions to the eyelid margin or surrounding areas provides therapeutic benefit to patients suffering from ocular surface disorders.

Claims

1-111. (canceled)

112. A compound having a structure represented by Formula (A-II): ##STR00226## wherein: X is an immunomodulator radical; each Y is independently a linker; each Z is independently a radical of a keratolytic agent; and n is 1-3; or a pharmaceutically acceptable salt or solvate thereof.

113. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein: n is 1 or 2; and X is selected from the group consisting of a radical of a phosphodiesterase (PDE) inhibitor, a radical of a Janus kinase (JAK) inhibitor, a radical of a folate reductase inhibitor, a radical of a steroid; and a radical of a calcineurin inhibitor.

114. The compound of claim 113, or a pharmaceutically acceptable salt or solvate thereof, wherein X is a radical of a JAK inhibitor.

115. The compound of claim 114, or a pharmaceutically acceptable salt or solvate thereof, wherein X is a radical of a JAK1, JAK2, and/or JAK3 inhibitor.

116. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein X has a structure represented by Formula (I-B): ##STR00227## wherein: R.sup.4 is hydrogen, halogen, or alkyl; R.sup.5 is hydrogen, halogen, or alkyl; R.sup.6 is hydrogen, halogen, or alkyl; R.sup.7 is NR.sup.bR.sup.c or optionally substituted heterocyclyl; and R.sup.b and R.sup.c are each independently hydrogen, alkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl.

117. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein each linker is the same, and is a bond, substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl.

118. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein: each linker is different, and is independently a bond, substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl.

119. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein each radical of a keratolytic agent is a radical of the same keratolytic agent, and comprises one or more groups, each group being independently selected from the group consisting of O, oxo, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocyclyl.

120. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein each radical of a keratolytic agent is a radical of a different keratolytic agent, and comprises one or more groups, each group being independently selected from the group consisting of O, oxo, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocyclyl.

121. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein each radical of a keratolytic agent is alkyl substituted with one or more substituent, each substituent being independently selected from the group consisting of oxo, hydroxy, alkyl, alkoxy, and substituted or unsubstituted heterocyclyl.

122. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein: each radical of a keratolytic agent is heteroalkyl substituted with one or more substituent, each substituent being independently selected from the group consisting of oxo, alkyl, thioalkyl, and substituted or unsubstituted heterocyclyl.

123. The compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, wherein each radical of a keratolytic agent has a structure represented by: ##STR00228## wherein: Q is O or (CR.sup.18R.sup.19).sub.m; m is 1-6; each R.sup.18 and R.sup.19 is independently H, halo, alkyl, alkoxy, haloalkyl, or thioalkyl; or an adjacent R.sup.18 and R.sup.19 combine to the atoms to which they are attached to form an oxo; and R.sup.20 is alkyl, heteroalkyl, heterocyclyl, alkoxy, or hydroxy, the alkyl, heteroalkyl, heterocycloalkyl, or alkoxy each independently being optionally substituted.

124. A compound having the structure: ##STR00229## ##STR00230## ##STR00231## ##STR00232## ##STR00233## ##STR00234## ##STR00235## ##STR00236## ##STR00237## ##STR00238## ##STR00239## ##STR00240## ##STR00241## ##STR00242## ##STR00243## ##STR00244## or a pharmaceutically acceptable salt or solvate thereof.

125. A compound having the structure: ##STR00245## ##STR00246## ##STR00247## ##STR00248## ##STR00249## ##STR00250## ##STR00251## or a pharmaceutically acceptable salt or solvate thereof.

126. A pharmaceutical composition comprising a compound of claim 112, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient.

127. A method of treating a dermal or an ocular disease or disorder in an individual, the method comprising administering a compound of claim 112 to the individual.

128. The method of claim 127, wherein the dermal or the ocular disease or disorder is associated with keratosis, microbial infiltration, microbial infection, inflammation, or any combination thereof.

Description

DETAILED DESCRIPTION OF THE DISCLOSURE

Certain Definitions

[0218] As used herein and in the appended claims, the singular forms a, and, and the include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to an agent includes a plurality of such agents, and reference to the cell includes reference to one or more cells (or to a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formulae, all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. The term about when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary between 1% and 15% of the stated number or numerical range. The term comprising (and related terms such as comprise or comprises or having or including) is not intended to exclude that in other certain embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, described herein, may consist of or consist essentially of the described features.

[0219] The terms treat, treating, or treatment as used herein, include reducing, alleviating, abating, ameliorating, managing, relieving, or lessening the symptoms associated with a disease, disease state, condition, or indication (e.g., provided herein) in either a chronic or acute therapeutic scenario. Also, treatment of a disease or disease state described herein includes the disclosure of use of such compound or composition for the treatment of such disease, disease state, disorder, or indication.

[0220] Amino refers to the NH.sub.2 radical.

[0221] Cyano refers to the CN radical.

[0222] Nitro refers to the NO.sub.2 radical.

[0223] Oxo refers to the O radical.

[0224] Hydroxyl refers to the OH radical.

[0225] Alkyl generally refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, such as having from one to fifteen carbon atoms (e.g., C.sub.1-C.sub.15 alkyl). Unless otherwise stated, alkyl is saturated or unsaturated (e.g., an alkenyl, which comprises at least one carbon-carbon double bond). Disclosures provided herein of an alkyl are intended to include independent recitations of a saturated alkyl, unless otherwise stated. Alkyl groups described herein are generally monovalent, but may also be divalent (which may also be described herein as alkylene or alkylenyl groups). In certain embodiments, an alkyl comprises one to thirteen carbon atoms (e.g., C.sub.1-C.sub.13 alkyl). In certain embodiments, an alkyl comprises one to eight carbon atoms (e.g., C.sub.1-C.sub.8 alkyl). In other embodiments, an alkyl comprises one to five carbon atoms (e.g., C.sub.1-C.sub.5 alkyl). In other embodiments, an alkyl comprises one to four carbon atoms (e.g., C.sub.1-C.sub.4 alkyl). In other embodiments, an alkyl comprises one to three carbon atoms (e.g., C.sub.1-C.sub.3 alkyl). In other embodiments, an alkyl comprises one to two carbon atoms (e.g., C.sub.1-C.sub.2 alkyl). In other embodiments, an alkyl comprises one carbon atom (e.g., C.sub.1 alkyl). In other embodiments, an alkyl comprises five to fifteen carbon atoms (e.g., C.sub.5-C.sub.15 alkyl). In other embodiments, an alkyl comprises five to eight carbon atoms (e.g., C.sub.5-C.sub.8 alkyl). In other embodiments, an alkyl comprises two to five carbon atoms (e.g., C.sub.2-C.sub.5 alkyl). In other embodiments, an alkyl comprises three to five carbon atoms (e.g., C.sub.3-C.sub.5 alkyl). In other embodiments, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl is attached to the rest of the molecule by a single bond. In general, alkyl groups are each independently substituted or unsubstituted.

[0226] Each recitation of alkyl provided herein, unless otherwise stated, includes a specific and explicit recitation of an unsaturated alkyl group. Similarly, unless stated otherwise specifically in the specification, an alkyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, OR.sup.a, SR.sup.a, OC(O)R.sup.a, N(R.sup.a).sub.2, C(O)R.sup.a, C(O)OR.sup.a, C(O)N(R.sup.a).sub.2, N(R.sup.a)C(O)OR.sup.a, OC(O)N(R.sup.a).sub.2, N(R.sup.a)C(O)R.sup.a, N(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2), S(O)tOR.sup.a (where t is 1 or 2), S(O).sub.tR.sup.a (where t is 1 or 2) and S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2) where each R.sup.a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0227] Alkoxy refers to a radical bonded through an oxygen atom of the formula O-alkyl, where alkyl is an alkyl chain as defined above.

[0228] Alkenyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and having from two to twelve carbon atoms. In certain embodiments, an alkenyl comprises two to eight carbon atoms. In other embodiments, an alkenyl comprises two to four carbon atoms. The alkenyl is optionally substituted as described for alkyl groups.

[0229] Alkylene or alkylene chain generally refers to a straight or branched divalent alkyl group linking the rest of the molecule to a radical group, such as having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, i-propylene, n-butylene, and the like. Unless stated otherwise specifically in the specification, an alkylene chain is optionally substituted as described for alkyl groups herein.

[0230] Aryl refers to a radical derived from an aromatic monocyclic or multicyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. The aromatic monocyclic or multicyclic hydrocarbon ring system contains only hydrogen and carbon from five to eighteen carbon atoms, where at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) -electron system in accordance with the Hckel theory. The ring system from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin and naphthalene. Unless stated otherwise specifically in the specification, the term aryl or the prefix ar- (such as in aralkyl) is meant to include aryl radicals optionally substituted by one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, R.sup.bOR.sup.a, R.sup.bOC(O)R.sup.a, R.sup.bOC(O)OR.sup.a, R.sup.bOC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a).sub.2, R.sup.bC(O)R.sup.a, R.sup.bC(O)OR.sup.a, R.sup.bC(O)N(R.sup.a).sub.2, R.sup.bOR.sup.cC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a)C(O)OR.sup.a, R.sup.bN(R.sup.a)C(O)R.sup.a, R.sup.bN(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tOR.sup.a (where t is 1 or 2) and R.sup.bS(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where each R.sup.a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), each R.sup.b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R.sup.c is a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0231] Aralkyl or aryl-alkyl refers to a radical of the formula R.sup.c-aryl where R.sup.c is an alkylene chain as defined above, for example, methylene, ethylene, and the like. The alkylene chain part of the aralkyl radical is optionally substituted as described above for an alkylene chain. The aryl part of the aralkyl radical is optionally substituted as described above for an aryl group.

[0232] Carbocyclyl or cycloalkyl refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused or bridged ring systems, having from three to fifteen carbon atoms. In certain embodiments, a carbocyclyl comprises three to ten carbon atoms. In other embodiments, a carbocyclyl comprises five to seven carbon atoms. The carbocyclyl is attached to the rest of the molecule by a single bond. Carbocyclyl or cycloalkyl is saturated (i.e., containing single CC bonds only) or unsaturated (i.e., containing one or more double bonds or triple bonds). Examples of saturated cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl is also referred to as cycloalkenyl. Examples of monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl radicals include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise stated specifically in the specification, the term carbocyclyl is meant to include carbocyclyl radicals that are optionally substituted by one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, R.sup.bOR.sup.a, R.sup.bOC(O)R.sup.a, R.sup.bOC(O)OR.sup.a, R.sup.bOC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a).sub.2, R.sup.bC(O)R.sup.a, R.sup.bC(O)OR.sup.a, R.sup.bC(O)N(R.sup.a).sub.2, R.sup.bOR.sup.cC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a)C(O)OR.sup.a, R.sup.bN(R.sup.a)C(O)R.sup.a, R.sup.bN(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tOR.sup.a (where t is 1 or 2) and R.sup.bS(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where each R.sup.a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), each R.sup.b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R.sup.c is a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0233] Carbocyclylalkyl refers to a radical of the formula R.sup.c-carbocyclyl where R.sup.c is an alkylene chain as defined above. The alkylene chain and the carbocyclyl radical is optionally substituted as defined above.

[0234] Carbocyclylalkenyl refers to a radical of the formula R.sup.c-carbocyclyl where R.sup.c is an alkenylene chain as defined above. The alkenylene chain and the carbocyclyl radical is optionally substituted as defined above.

[0235] Carbocyclylalkoxy refers to a radical bonded through an oxygen atom of the formula OR.sup.c-carbocyclyl where R.sup.c is an alkylene chain as defined above. The alkylene chain and the carbocyclyl radical is optionally substituted as defined above.

[0236] Halo or halogen refers to fluoro, bromo, chloro, or iodo substituents.

[0237] Haloalkyl refers to an alkyl radical, as defined above, that is substituted by one or more halogen radicals, as defined above, for example, trihalomethyl, dihalomethyl, halomethyl, and the like. In some embodiments, the haloalkyl is a fluoroalkyl, such as, for example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. In some embodiments, the alkyl part of the fluoroalkyl radical is optionally substituted as defined above for an alkyl group.

[0238] The term heteroalkyl refers to an alkyl group as defined above in which one or more skeletal carbon atoms of the alkyl are substituted with a heteroatom (with the appropriate number of substituents or valenciesfor example, CH.sub.2 may be replaced with NH or O). For example, each substituted carbon atom is independently substituted with a heteroatom, such as wherein the carbon is substituted with a nitrogen, oxygen, sulfur, or other suitable heteroatom. In some instances, each substituted carbon atom is independently substituted for an oxygen, nitrogen (e.g. NH, N(alkyl)-, or N(aryl)- or having another substituent contemplated herein), or sulfur (e.g. S, S(O), or S(O).sub.2). In some embodiments, a heteroalkyl is attached to the rest of the molecule at a carbon atom of the heteroalkyl. In some embodiments, a heteroalkyl is attached to the rest of the molecule at a heteroatom of the heteroalkyl. In some embodiments, a heteroalkyl is a C.sub.1-C18 heteroalkyl. In some embodiments, a heteroalkyl is a C.sub.1-C.sub.12 heteroalkyl. In some embodiments, a heteroalkyl is a C.sub.1-C.sub.6 heteroalkyl. In some embodiments, a heteroalkyl is a C.sub.1C.sub.4 heteroalkyl. Representative heteroalkyl groups include, but are not limited to OCH.sub.2OMe, or CH.sub.2CH.sub.2OMe. In some embodiments, heteroalkyl includes alkoxy, alkoxyalkyl, alkylamino, alkylaminoalkyl, aminoalkyl, heterocycloalkyl, heterocycloalkyl, and heterocycloalkylalkyl, as defined herein. Unless stated otherwise specifically in the specification, a heteroalkyl group is optionally substituted as defined above for an alkyl group.

[0239] Heteroalkylene refers to a divalent heteroalkyl group defined above which links one part of the molecule to another part of the molecule. Unless stated specifically otherwise, a heteroalkylene is optionally substituted, as defined above for an alkyl group.

[0240] Heterocyclyl refers to a stable 3- to 18-membered non-aromatic ring radical that comprises two to twelve carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. Unless stated otherwise specifically in the specification, the heterocyclyl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which optionally includes fused or bridged ring systems. The heteroatoms in the heterocyclyl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heterocyclyl radical is partially or fully saturated. The heterocyclyl radical is saturated (i.e., containing single CC bonds only) or unsaturated (e.g., containing one or more double bonds or triple bonds in the ring system). In some instances, the heterocyclyl radical is saturated (e.g., dithiolanyl or dithiolanyl oxide). In some instances, the heterocyclyl radical is saturated and substituted (e.g., dithiolanyl oxide). In some instances, the heterocyclyl radical is unsaturated. The heterocyclyl is attached to the rest of the molecule through any atom of the ring(s). Examples of such heterocyclyl radicals include, but are not limited to, dithiolanyl, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, the term heterocyclyl is meant to include heterocyclyl radicals as defined above that are optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, R.sup.bOR.sup.a, R.sup.bOC(O)R.sup.a, R.sup.bOC(O)OR.sup.a, R.sup.bOC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a).sub.2, R.sup.bC(O)R.sup.a, R.sup.bC(O)OR.sup.a, R.sup.bC(O)N(R.sup.a).sub.2, R.sup.bOR.sup.cC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a)C(O)OR.sup.a, R.sup.bN(R.sup.a)C(O)R.sup.a, R.sup.bN(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tOR.sup.a (where t is 1 or 2) and R.sup.bS(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where each R.sup.a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), each R.sup.b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R.sup.c is a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0241] N-heterocyclyl or N-attached heterocyclyl refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical. An N-heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals.

[0242] Examples of such N-heterocyclyl radicals include, but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

[0243] C-heterocyclyl or C-attached heterocyclyl refers to a heterocyclyl radical as defined above containing at least one heteroatom and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a carbon atom in the heterocyclyl radical. A C-heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals.

[0244] Examples of such C-heterocyclyl radicals include, but are not limited to, 2-morpholinyl, 2- or 3-or 4-piperidinyl, 2-piperazinyl, 2- or 3-pyrrolidinyl, and the like.

[0245] Heterocyclylalkyl refers to a radical of the formula R.sup.c-heterocyclyl where R.sup.c is an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain of the heterocyclylalkyl radical is optionally substituted as defined above for an alkylene chain. The heterocyclyl part of the heterocyclylalkyl radical is optionally substituted as defined above for a heterocyclyl group.

[0246] Heterocyclylalkoxy refers to a radical bonded through an oxygen atom of the formula OR.sup.c-heterocyclyl where R.sup.c is an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain of the heterocyclylalkoxy radical is optionally substituted as defined above for an alkylene chain. The heterocyclyl part of the heterocyclylalkoxy radical is optionally substituted as defined above for a heterocyclyl group.

[0247] Heteroaryl refers to a radical derived from a 3- to 18-membered aromatic ring radical that comprises two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. As used herein, the heteroaryl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) -electron system in accordance with the Hckel theory. Heteroaryl includes fused or bridged ring systems. The heteroatom(s) in the heteroaryl radical is optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heteroaryl is attached to the rest of the molecule through any atom of the ring(s).

[0248] Examples of heteroaryls include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e. thienyl). Unless stated otherwise specifically in the specification, the term heteroaryl is meant to include heteroaryl radicals as defined above which are optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, R.sup.bOR.sup.a, R.sup.bOC(O)R.sup.a, R.sup.bOC(O)OR.sup.a, R.sup.bOC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a).sub.2, R.sup.bC(O)R.sup.a, R.sup.bC(O)OR.sup.a, R.sup.bC(O)N(R.sup.a).sub.2, R.sup.bOR.sup.cC(O)N(R.sup.a).sub.2, R.sup.bN(R.sup.a)C(O)OR.sup.a, R.sup.bN(R.sup.a)C(O)R.sup.a, R.sup.bN(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tR.sup.a (where t is 1 or 2), R.sup.bS(O).sub.tOR.sup.a (where t is 1 or 2) and R.sup.bS(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where each R.sup.a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), each R.sup.b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R.sup.c is a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0249] N-heteroaryl refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical. An N-heteroaryl radical is optionally substituted as described above for heteroaryl radicals.

[0250] C-heteroaryl refers to a heteroaryl radical as defined above and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a carbon atom in the heteroaryl radical. A C-heteroaryl radical is optionally substituted as described above for heteroaryl radicals.

[0251] Heteroarylalkyl refers to a radical of the formula R.sup.c-heteroaryl, where R.sup.c is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain of the heteroarylalkyl radical is optionally substituted as defined above for an alkylene chain. The heteroaryl part of the heteroarylalkyl radical is optionally substituted as defined above for a heteroaryl group.

[0252] Heteroarylalkoxy refers to a radical bonded through an oxygen atom of the formula OR.sup.c-heteroaryl, where R.sup.c is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain of the heteroarylalkoxy radical is optionally substituted as defined above for an alkylene chain. The heteroaryl part of the heteroarylalkoxy radical is optionally substituted as defined above for a heteroaryl group.

[0253] The compounds disclosed herein, in some embodiments, contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that are defined, in terms of absolute stereochemistry, as (R)- or (S)-. Unless stated otherwise, it is intended that all stereoisomeric forms of the compounds disclosed herein are contemplated by this disclosure. When the compounds described herein contain alkene double bonds, and unless specified otherwise, it is intended that this disclosure includes both E and Z geometric isomers (e.g., cis or trans.) Likewise, all possible isomers, as well as their racemic and optically pure forms, and all tautomeric forms are also intended to be included. The term geometric isomer refers to E or Z geometric isomers (e.g., cis or trans) of an alkene double bond. The term positional isomer refers to structural isomers around a central ring, such as ortho-, meta-, and para-isomers around a benzene ring.

[0254] In general, optionally substituted groups are each independently substituted or unsubstituted. Each recitation of an optionally substituted group provided herein, unless otherwise stated, includes an independent and explicit recitation of both an unsubstituted group and a substituted group (e.g., substituted in certain embodiments, and unsubstituted in certain other embodiments). Unless otherwise stated, a substituted group provided herein (e.g., substituted alkyl) is substituted by one or more substituent, each substituent being independently selected from the group consisting of halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, OR.sup.a, SR.sup.a, OC(O)R.sup.a, N(R.sup.a).sub.2, C(O)R.sup.a, C(O)OR.sup.a, C(O)N(R.sup.a).sub.2, N(R.sup.a)C(O)OR.sup.a, OC(O)N(R.sup.a).sub.2, N(R.sup.a)C(O)R.sup.a, N(R.sup.a)S(O).sub.tR.sup.a (where t is 1 or 2), S(O).sub.tOR.sup.a (where t is 1 or 2), S(O).sub.tR.sup.a (where t is 1 or 2) and S(O).sub.tN(R.sup.a).sub.2 (where t is 1 or 2), where each R.sup.a is independently hydrogen, alkyl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (e.g., optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0255] Pharmaceutically acceptable salt includes both acid and base addition salts. A pharmaceutically acceptable salt of any one of the pharmacological agents described herein is intended to encompass any and all pharmaceutically suitable salt forms. Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0256] Pharmaceutically acceptable acid addition salt refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, and the like. Also included are salts that are formed with organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and. aromatic sulfonic acids, etc. and include, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Exemplary salts thus include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, trifluoroacetates, propionates, caprylates, isobutyrates, oxalates, malonates, succinate suberates, sebacates, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like.

[0257] Also contemplated are salts of amino acids, such as arginates, gluconates, and galacturonates (see, for example, Berge S. M. et al., Pharmaceutical Salts, Journal of Pharmaceutical Science, 66:1-19 (1997)). Acid addition salts of basic compounds are, in some embodiments, prepared by contacting the free base forms with a sufficient amount of the desired acid to produce the salt according to methods and techniques with which a skilled artisan is familiar.

[0258] Pharmaceutically acceptable base addition salt refers to those salts that retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Pharmaceutically acceptable base addition salts are, in some embodiments, formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. See Berge et al., supra.

[0259] The term keratinized obstruction, as used herein, generally refers to a blockage of the meibomian gland, regardless of the location of the blockage. In some embodiments, the blockage is complete, whereas in other embodiments, the blockage is partial. Regardless of the degree of blockage, such keratinized obstruction leads to meibomian gland dysfunction. In some embodiments, the keratinized obstruction is composed of keratinized material and lipids. In some embodiments, the keratinized obstruction is a blockage at the meibomian gland orifice and excretory duct. In some embodiments, the keratinized obstruction is caused by keratinization of the epithelium at the lid margin and meibomian gland. In certain instances, the keratin obstruction is influenced by the migration or aberrant differentiation of stem cells. In some embodiments, the keratinized obstruction results in reduced delivery of oil to the lid margin and tear film, and stasis inside the meibomian gland that causes increased pressure, resultant dilation, acinar atrophy, and low secretion. In certain instances, keratinization of the meibomian gland causes degenerative gland dilation and atrophy.

[0260] The term, meibomian gland dysfunction, as used herein, refers to chronic, diffuse abnormality of the meibomian glands, that is characterized by terminal duct obstruction or qualitative or quantitative changes in the glandular secretion, or both. MGD may result in alteration of the tear film, eye irritation symptoms, inflammation, or ocular surface disease. The most prominent aspects of MGD are obstruction of the meibomian gland orifices and terminal ducts and changes in the meibomian gland secretions.

[0261] The meibomian glands are large sebaceous glands located in the eyelids, and unlike skin, are unassociated with hair. The meibomian glands produce the lipid layer of the tear film that protects it against evaporation of the aqueous phase. The meibomian gland orifice is located on the epithelial side of the lid margin, and can be a few hundred microns from the mucosal side. The glands are located on both upper and lower eyelids, with higher amounts of the glands on the upper eyelid. A single meibomian gland is composed of clusters of secretory acini that are arranged circularly around a long central duct and connected to it by short ductules. The terminal part of the central duct is lined by an ingrowth of the epidermis that covers the free lid margin and forms a short excretory duct that opens as an orifice at the posterior part of the lid margin just anterior to the mucocutaneous junction near the inner lid border. The oily secretion composed of lipids is synthesized within the secretory acini. The lipid secretion is a liquid at near body temperature and is delivered to the skin of the lid margin as a clear fluid, called meibum. It forms shallow reservoirs on the upper and lower lid margins, and consists of a complex mixture of cholesterol, wax, cholesteryl esters, phospholipids, with small amounts of triglycerides, triacylglycerols, and hydrocarbons. The separate meibomian glands are arranged in parallel, and in a single row throughout the length of the tarsal plates in the upper and lower lids. The extent of the glands corresponds roughly to the dimensions of the tarsal plates.

[0262] Ocular surface diseases are a group of diseases including, but not limited to, dry eye syndrome (including evaporative DES and/or aqueous deficiency DES), blepharitis, keratitis, meibomian gland dysfunction, conjunctivitis, lacrimal gland disorder, contact lens related conditions and inflammatory, infectious, or autoimmune diseases or disorders of the anterior surface of the eye.

[0263] In some instances, meibomian gland dysfunction (MGD) is a chronic, diffuse abnormality of the meibomian glands, which can be characterized by terminal duct obstruction and/or qualitative/quantitative changes in the glandular secretion. Terminal duct obstruction is caused by hyperkeratinization of the ductal epithelium (Nichols et al, Inv. Oph. & Vis. Sci. (2011); 52(4):1922-1929). These alterations in both meibum quality and expression may result in alteration of the tear film, symptoms of eye irritation, and ocular surface disease such as evaporative dry eye. The principal clinical consequence of MGD is evaporative dry eye syndrome and large population based studies (i.e., Bankok Study and the Shihpai Eye Study) estimate that over 60% of patients with dry eye symptoms also have MGD (Schaumberg et al, Investigative Ophthalmology and Visual Science. (2011); 52(4):1994-2005).

[0264] MGD is a leading contributor of dry eye syndrome. The occurrence of dry eye syndrome is widespread and affects about 20 million patients in the United States alone. Dry eye syndrome is a disorder of the ocular surface resulting from either inadequate tear production or excessive evaporation of moisture from the surface of the eye. Tears are important to corneal health because the cornea does not contain blood vessels, and relies on tears to supply oxygen and nutrients. Tears and the tear film are composed of lipids, water, and mucus, and disruption of any of these can cause dry eye. An inadequate amount of lipids flowing from the meibomian glands as caused by a keratinized obstruction, may cause excessive evaporation, thereby causing dry eye syndrome.

[0265] Currently there are no approved pharmacological agents useful for the treatment of MGD. The recognition that terminal duct obstruction from hyperkeratinization of the ductal epithelium on meibomian glands is a core mechanism behind meibomian gland dysfunction (MGD) is consistent with clinical experience demonstrating that effective treatments for MGD require resolution of ductal obstruction and evacuation of glandular contents (Nichols et al, 2011; Lane et al, 2012; Blackie et al, 2015). Warm compresses and thermal/mechanical devises (e.g., LipiFlow) are used in an attempt to raise the internal temperature of the meibomian glands over the normal melting point for meibum (i.e., 32 C. to 40 C.) in an attempt to resolve terminal duct obstruction (Lane et al, 2012). Unfortunately, warm compresses are unable to achieve this benefit for severely obstructed glands which can having a melting point >40 C. Current technology for removing keratinized obstruction of the meibomian gland also includes physical removal methods (e.g., debridement and gland probing), which are quite painful to patients.

[0266] Subsequent to a period of MGD, various stages of immune, inflammatory, and/or bacterial disease at the ocular surface are frequently observed because meibomian gland obstruction can cause a cascade of events that include further deterioration of the glands (Knop, IOVS, 2011) from stasis of the meibum in the secretory glands, mechanical pressure and stress from glandular obstruction, and increased bacterial growth that is associated with the downstream release of bacterial lipases, toxic mediators, and/or inflammatory mediators. All these factors reduce the quality and/or quantity of meibum the glands can release which in turn can cause chronic mechanical traumatization of the conjunctival, corneal and eyelid tissues which will lead to further tissue damage and the release of inflammatory mediators. Thus, many patients suffering from MGD also have inflammatory disease affecting their conjunctiva, cornea, larcrimal gland, lids or goblet cells causing comorbid conditions such as dry eye syndrome or blepharitis for which there is an unmet medical need.

[0267] For example, literature has used the terms posterior blepharitis and MGD as if they were synonymous, but these terms are not interchangeable. Posterior blepharitis describes inflammatory conditions of the posterior lid margin, of which MGD can be one possible cause. In its earliest stages, MGD may not be associated with clinical signs characteristic of posterior blepharitis. At this stage, affected individuals may be symptomatic, but alternatively, they may be asymptomatic and the condition regarded as subclinical. As MGD progresses, symptoms develop and lid margin signs, such as changes in meibum expressibility and quality and lid margin redness, may become more visible. At this point, an MGD-related posterior blepharitis is said to be present.

[0268] In some instances, altered meibomian gland secretion is detected by physically expressing the meibomian glands by applying digital pressure to the tarsal plates. In subjects without MGD, the meibum is a pool of clear oil. In MGD, both the quality and expressibility of the expressed material is altered. The altered meibum is also known as meibomian excreta and is made up of a mixture of altered secretions and keratinized epithelial material. In MGD, the quality of expressed lipid varies in appearance from a clear fluid, to a viscous fluid containing particulate matter and densely opaque, toothpaste-like material. The meibomian orifices may exhibit elevations above surface level of the lid, which is referred to as plugging or pouting, and is due to obstruction of the terminal ducts and extrusion of a mixture of meibomian lipid and keratinized material.

[0269] In some instances, obstructive MGD is characterized by all or some of the following: 1) chronic ocular discomfort, 2) anatomic abnormalities around the meibomian gland orifice (which is one or more of the following: vascular engorgement, anterior or posterior displacement of the mucocutaneous junction, irregularity of the lid margin) and 3) obstruction of the meibomian glands (obstructive findings of the gland orifices by slit lamp biomicroscopy (pouting, plugging or ridge), decreased meibum expression by moderate digital pressure).

[0270] Methods for assessing and monitoring MGD symptoms may include, but are not limited to patient questionnaires, meibomian gland expression, tear stability break up time, and determining the number of patent glands as seen by digital expression.

[0271] In some embodiments, the symptoms of a patient are assessed by asking the patient a series of questions. Questionnaires allow the assessment of a range of symptoms associated with ocular discomfort. In some embodiments, the questionnaire is the SPEED questionnaire. The SPEED questionnaire assesses frequency and severity of a patient's dry eye symptoms. It examines the occurrence of symptoms on the current day, past 72 hours and past three months. A SPEED score is tallied based on the patient's answers to the questions, to give a range of severity of the patient's symptoms. The SPEED questionnaire includes questions such as the following: 1) what dry eye symptoms are you experiencing, and when do they occur?2) how frequently do you experience dryness, grittiness, or scratchiness in your eyes?3) how often do you experience soreness or irritation of the eyes?4) how often do you experience burning or watering of the eyes?5) how often do you experience eye fatigue? and 6) how severe are the symptoms?

[0272] Meibomian gland expressibility is optionally determined to assess the meibomian gland function. In normal patients, meibum is a clear to light yellow oil. Meibum is excreted from the glands when digital pressure is placed on the glands. Changes in meibomian gland expressibility are one potential indicator of MGD. In some embodiments, during expression, quantifying the amount of physical force applied during expression is monitored in addition to assessing lipid volume and lipid quantity.

[0273] Tear stability break up time (TBUT) is a surrogate marker for tear stability. Tear film instability is a core mechanism in dry eye and MGD. Low TBUT implies a possibility of lipid layer compromise and MGD. TBUT is optionally measured by examining fluorescein breakup time, as defined as the time to initial breakup of the tear film after a blink. Fluorescein is optionally applied by wetting a commercially available fluorescein-impregnated strip with saline, and applied to the inferior fornix or bulbar conjuctiva. The patient is then asked to blink several times and move the eyes. The break up is then analyzed with a slit lamp, a cobalt blue filter, and a beam width of 4 mm. The patient is instructed to blink, and the time from upstroke of the last blink to the first tear film break or dry spot formation is recorded as a measurement.

[0274] Other methods for assessing MGD symptoms, include but are not limited to, Schirmer test, ocular surface staining, lid morphology analysis, meibography, meibometry, interferometry, evaporimetry, tear lipid composition analysis, fluorophotometry, meiscometry, osmolarity analysis, indices of tear film dynamics, evaporation and tear turnover.

[0275] Treatments for MGD can include lid warming, lid massage, lid hygiene, lid expression and meibomian gland probing. Pharmacological methods, prior to those described herein, have not been used.

[0276] Lid hygiene is considered the primary treatment for MGD and consists of three components: 1) application of heat, 2) mechanical massage of eyelids and 3) cleansing the eyelid. Eyelid warming procedures improve meibomian gland secretion by melting the pathologically altered meibomian lipids. Warming is achieved by warm compresses or devices. Mechanical lid hygiene includes the use of scrubs, mechanical expression and cleansing with various solutions of the eyelashes and lid margins. Lid margins are optionally also cleansed with hypoallergenic bar soap, dilute infant shampoo or commercial lid scrubs. Physical expression of meibomian glands is performed in a physician's office or is performed by the patient at home. The technique varies from gentle massage of the lids against the eyeball to forceful squeezing of the lids either against each other or between a rigid object on the inner lid surface and a finger, thumb, or rigid object (such as a glass rod, cotton swab, or metal paddle) on the outer lid surface. The rigid object on the inner lid surface protects the eyeball from forces transferred through the eyelid during expression and to offer a stable resistance, to increase the amount of force that is applied to the glands.

[0277] Eyelid warming is limited because the warming melts the lipids, but does not address movement of the keratinized material. Further, eyelid warming induces transient visual degradation due to corneal distortion. Mechanical lid hygiene is also limited because the force needed to remove an obstruction can be significant, resulting in significant pain to the patient. The effectiveness of mechanical lid hygiene is limited by the patient's ability to tolerate the associated pain during the procedure. Other treatments for MGD are limited.

[0278] Physical opening of meibomian glands obstruction by meibomian gland expression is an acceptable method to improve meibomian gland secretion and dry eye symptoms. In addition, probing of the meibomian gland canal has been used to open the obstructed canal. Both methods, expression and probing, are limited, however, by the pain induced by the procedure, the possible physical insult to the gland and canal structures and their short lived effect estimated at days and weeks. Therefore, methods are needed to improve patient comfort, which will not cause harm to the meibomian glands and canals, that will reduce the dependency on frequent office visits and improve secretion of meibum.

[0279] Patent U.S. Pat. No. 9,463,201 entitled, Compositions and methods for the treatment of meibomian gland dysfunction describes a method for treating meibomian gland dysfunction involving the topical administration of a therapeutically-effective amount of at least one keratolytic agent in an ophthalmically-acceptable carrier. The patent includes keratolytic agents that are inorganic selenium (Se) compounds such as selenium disulfide (SeS.sub.2) or organoselenium compounds such as Ebselen (2-Phenyl-1,2-benzoselenazol-3-one). This agent would treat the underlying cause of MGD, but not a plus inflammatory disease as described by the DEWS report on MGD.

[0280] The role of inflammation in the etiology of MGD is controversial. The terms posterior blepharitis and MGD are not synonymous. Posterior blepharitis describes inflammatory conditions of the posterior lid margin and has various causes, of which MGD can be one possible cause (Nichols et al 2011). In its earliest stages, MGD is not associated with clinical signs characteristic of posterior blepharitis. As MGD progresses, an MGD-related posterior blepharitis is said to be present. MGD-related posterior blepharitis affects the meibomian glands and meibomian gland orifices. MGD-related posterior blepharitis is characterized by flora changes, esterase and lipase release, lipid changes, and eyelid inflammation. Hyperkeratinization of the meibomian gland epithelium (thickening of the lining of the glands) may lead to obstruction and a decrease in the quantity of meibomian gland secretions and may be responsible for MGD-related posterior blepharitis. Diagnosis of MGD-related posterior blepharitis includes meibomian gland expression with demonstration of an altered quality of expressed secretions, and/or by a loss of gland functionality (decreased or absent expressibility). The TFOS report on Meibomian Gland Disease specifically notes that anterior blepharitis and exacerbated inflammatory ocular surface disease are plus diseases to MGD which are managed by topical, ocular steroids (Nichols et al 2011).

[0281] Since these plus conditions can be present in various levels of severity from early to late MGD there is a need for treatments and/or combinations of treatments that can target both the underlying non-inflammatory pathophysiology of MGD and inflammation associated with these comorbid conditions.

[0282] MGD-related inflammatory eye disease may comprise a different mechanism than blepharitis-related MGD. MGD-related inflammatory eye disease is characterized by an inflammatory cascade involving activation and migration of T lymphocytes to the inflamed tissue. T lymphocyte infiltration may result in lacrimal gland stimulation and upregulation of cytokines. Exemplary cytokines that may be involved in MGD-related inflammatory eye disease include, but are not limited to, interleukin-1, interleukin-4, interleukin-6, inteleukin-8, interferon gamma, macrophage inflammatory protein 1 alpha, and tumor necrosis factor alpha. Kinase pathways including the mitogen activated protein kinase (MAPK) pathway are also activated in the inflammatory cascade. The inflammatory process results in loss of mucin-producing goblet cells and destruction of the ocular surface that can lead to further damage.

[0283] Dry eye syndrome, also known as keratoconjunctivitis sicca (KCS), is considered a self-sustaining disease that is progressively disconnected from its initial cause. Dry eye syndrome is associated with inflammation at the ocular surface and periocular tissue. Inflammation is characterized by the activation and migration of T lymphocytes to the inflamed tissue including in the conjunctiva and lacrimal glands. Inflammatory cytokines, chemokines, and matrix metalloproteinase have also been identified as being increased.

[0284] Animal models of dry eye disease have been established and reviewed (Barabino, et al, (Invest. Ophthalmol. Vis. Sci. 2004, 45:1641-1646)). Barabino, et al, (Invest. Ophthalmol. Vis. Sci. 2005, 46:2766-2771) described a model wherein exposure of normal mice to a low-humidity environment in a controlled-environment chamber leads to significant alterations in tear secretion, goblet cell density, and acquisition of dry eye-related ocular surface signs. However, no single animal model adequately accounts for the immune, endocrine, neuronal and environmental factors which contribute to dry eye pathogenesis.

[0285] Anti-inflammatory agents may be used to treat ocular surface diseases or disorders including dry eye syndrome. Corticosteroids are an effective anti-inflammatory therapy in dry eye disease. For example, in a 4-week, double-masked, randomized study in 64 patients with dry eye and delayed tear clearance, loteprednol etabonate 0.5% ophthalmic suspension (Lotemax [Bausch and Lomb, Rochester, NY]), QID, was found to be more effective than its vehicle in improving some signs and symptoms (Pflugfelder et al, Am J Ophthalmol (2004); 138:444-57). The TFOS 2007 report on dry eye disease went so far as to conclude that, In the US Federal Regulations, ocular corticosteroids receiving class labeling are indicated for the treatment . . . of steroid responsive inflammatory conditions of the palpebral and bulbar conjunctiva, cornea and anterior segment of the globe such as allergic conjunctivitis, acne rosacea, superficial punctate keratitis, herpes zoster keratitis, iritis, cyclitis, selected infective conjunctivitis, when the inherent hazard of steroid use is accepted to obtain an advisable diminution in edema and inflammation. KCS, in some instances, is included in this list of steroid-responsive inflammatory conditions (Therapy Subcommittee of the International Dry Eye WorkShop, 2007. Management and Therapy of Dry Eye Disease: Report of the Management and Therapy Subcommittee of the International Dry Eye WorkShop (2007). 2007; 5: 163-178). While the US FDA does not agree with this conclusion, short courses of steroids, especially Lotemax, can be commonly used to treat inflammation associated with dry eye disease.

[0286] Other anti-inflammatory agents include nonsteroidal anti-inflammatory drugs (NSAIDs).

[0287] NSAIDs inhibit the activity of cyclooxygenases including cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2), which are enzymes involved in the synthesis of prostaglandins and thromboxanes from arachidonic acid. Prostaglandin and thromboxane signaling are involved in inflammation and immune modulation. In some cases, NSAIDs are used for treating dry eye disease by treating the inflammation at the ocular surface.

[0288] In some instances, provided herein is a compound that provides a therapeutically effective amount of (e.g., a free form of) an immunomodulator, such as an immunomodulator described herein, and/or (e.g., a free form of) a keratolytic agent, such as a keratolytic agent described herein.

[0289] In some instances, the free form of the immunomodulator is selected from the group consisting of cilomilast, ruxolitinib, ritlecitinib, tofacitinib, oclacitinib, methotrexate, loteprednol, and tacrolimus.

[0290] The chemical name for cilomilast is cis-4-cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]cyclohexanecarboxylic acid having a molecular formula of C.sub.20H.sub.25NO.sub.4 and a molecular weight of 343.4 g/mol. The structural formula of cilomilast is:

##STR00033##

[0291] Cilomilast is a potent and selective phosphodiesterase-4 (PDE4) inhibitor that has demonstrated improvements in objective signs of dry eye in a murine model (Sadrai et al, Invest Ophthalmol Vis Sci. (2012) 53 (7), 3584-3591). The potent and selective phosphodiesterase-4 (PDE4) inhibitor was found to act locally at the level of the ocular surface, by suppressing the generation of IL-17-associated immunity.

[0292] The chemical name for ruxolitinib is (3R)-3-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propanenitrile having a molecular formula of C.sub.17H.sub.18N.sub.6 and a molecular weight of 306.4 g/mol. The structural formula of ruxolitinib is:

##STR00034##

[0293] Ruxolitinib is a potent and selective janus kinase (JAK) 1 and JAK2 inhibitor that has demonstrated improvements in objective signs (in murine models) of psoriasis (Mesa, IDrugs. (2010) 13 (6), 394-403), alopecia (Falto-Aizpurua et al, Expert Opinion on Emerging Drugs. (2014) 19 (4), 545-556), and uveitis (caused by Salmonella typhiurium endotoxin) (Lin et al, Microorganisms (2021) 9 (7), 1-12). The potent and selective JAK1 and JAK2 inhibitor was found to act locally at the level of the ocular surface, possibly by suppressing the expression of mediators of proinflammatory cytokine pathways (e.g., of the JAK2-signal transducers and activators of transcription 3 (STAT3) pathway) in the ciliary body and iris.

[0294] The chemical name for ritlecitinib is 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one having a molecular formula of C.sub.15H.sub.19N.sub.5O and a molecular weight of 285.3 g/mol. The structural formula of ritlecitinib is:

##STR00035##

[0295] Ritlecitinib is an irreversible covalent JAK3 selective inhibitor that has demonstrated improvements in objective signs (in humans and murine models) of alopecia (NCT02974868). The irreversible covalent JAK3 selective inhibitor was found to inhibit the expression of mediators of proinflammatory cytokine pathways (such as IL-7, IL-9, IL-15 and IL-21), some of which being implicated in the pathophysiology of ocular indications, such as, dry eye disease and uveitis.

[0296] The chemical name for tofacitinib is 3-[(3R,4R)-4-Methyl-3-[methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino]piperidin-1-yl]-3-oxopropanenitrile having a molecular formula of C.sub.16H.sub.20N.sub.6O and a molecular weight of 312.4 g/mol. The structural formula of tofacitinib is:

##STR00036##

[0297] Tofacitinib is a potent and selective JAK1 and JAK3 inhibitor that has demonstrated improvements in objective signs (in humans and/or murine models) of psoriasis (Di Lernia et al, Drug Design, Development and Therapy. (2016) 10, 533-539), alopecia (Kennedy Crispin et al, JCI Insight. (2016) 1 (15), e89776), dermatitis (Levy et al, Journal of the American Academy of Dermatology (2015) 73 (3), 395-399), and dry eye disease (Jing-Feng Huang, US Ophthalmic Review. (2014) 7(1), 12-15). The potent and selective JAK1 and JAK3 inhibitor was found to act locally at the level of the ocular surface, possibly by suppressing the expression of mediators of proinflammatory cytokine pathways (e.g., IFN-7, IL-12, IL-23, and IL-6).

[0298] The chemical name for oclacitinib is N-methyl{trans-4-[methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino]cyclohexyl}methanesulfonamide having a molecular formula of C.sub.15H.sub.23N.sub.5O.sub.2S and a molecular weight of 337.4. The structural formula of oclacitinib is:

##STR00037##

[0299] Oclacitinib is a potent and selective JAK1, JAK2, and JAK3 inhibitor that has demonstrated improvements in objective signs (in humans and/or dogs) of dermatitis (Gonzalez et al, Journal of Veterinary Pharmacology and Therapeutics (2014) 37 (4), 317-324), keratoconjunctivitis, and dry eye disease (Kravetz de Oliveira, Vet Ophthalmol. (2019).sub.22(5), 633-643). The potent and selective JAK1, JAK2, and JAK3 inhibitor was found to act locally at the level of the ocular surface, possibly by suppressing the expression of mediators of proinflammatory cytokine pathways (e.g., IL-2, IL-4, IL-6, IL-13, and IL-31).

[0300] The chemical name for methotrexate is (2S)-2-[(4-{[(2,4-Diaminopteridin-6-yl)methyl](methyl)amino}benzoyl)amino]pentanedioic acid having a molecular formula of C.sub.20H.sub.22N.sub.8O.sub.5 and a molecular weight of 454.4 g/mol. The structural formula of methotrexate is:

##STR00038##

[0301] While prophylactic methotrexate has been associated with ocular side effect, such as, eye irritation and dry eye, methotrexate has demonstrated improvements in objective signs (in humans and/or murine models) of uveitis (Gangaputra, Ophthalmology (2009) 116(11), 2188-2198). Methotrexate was found to act locally at the level of the ocular surface, possibly by suppressing and/or activating the expression of mediators of certain immunological pathways.

[0302] The chemical name for loteprednol is chloromethyl 17-ethoxycarbonyloxy-11-hydroxy-10,13-dimethyl-3-oxo-7,8,9,11,12,14,15,16-octahydro-6H-cyclopenta[a]phenanthrene-17-carboxylate having a molecular formula of C.sub.24H.sub.31ClO.sub.7 and a molecular weight of 467.0 g/mol. The structural formula of loteprednol is:

##STR00039##

[0303] Loteprednol has demonstrated improvements in objective signs (in humans) of anterior segment inflammatory conditions of the eye, such as, for example, ocular inflammation, conjunctivitis, uveitis, keratitis, keratoconjunctivitis, and the like (Bartlett et al, US Ophthalmic Review. (2011) 4(1), 57-62). Loteprednol was found to act locally at the level of the ocular surface, possibly by suppressing and/or activating the expression of mediators of certain immunological pathways.

[0304] The chemical name for tacrolimus is ()-(3S,4R,5S,8R,9E,12S,14S,15R,16S,18R,26aS)-8-allyl-5,6,8,11,12,13,14,15,16,17,18,19,24,25,26,26a-hexadecahydro-5,19-dihydroxy-3-{(E)-2-[(1R,3R,4R)-4-hydroxy-3-methylcyclohexyl]-1-methylvinyl}-14,16-dimethoxy-4,10,12,18-tetramethyl-15,19-epoxy-3H-pyrido[2,1-c][1,4]oxaazacyclotricosane-1,7,20,21(4H,23H)-tetrone having a molecular formula of C.sub.44H.sub.69NO.sub.12 and a molecular weight of 804.0 g/mol. The structural formula of tacrolimus is:

##STR00040##

[0305] Tacrolimus is a calcineurin inhibitor that has demonstrated improvements in objective signs (in humans and/or dogs) of keratoconjunctivitis and dry eye disease (Kravetz de Oliveira, Vet Ophthalmol. (2019) 22(5), 633-643). The potent and selective JAK1, JAK2, and JAK3 inhibitor was found to act locally at the level of the ocular surface, possibly by interfering with interleukin-2 transcription, reducing lacrimal gland inflammation, and/or restoring corneal health.

Compositions

[0306] Described herein are compounds (e.g., keratolytic conjugates and/or dual acting-agents) which address simultaneously the non-inflammatory keratosis (e.g., keratolytic blockage) component of dermal and/or ocular diseases or disorders described herein (e.g., MGD) and the improper immunomodulation and/or inflammation associated with the disease or disorder described herein (e.g., an ocular allergy, dry eye disease (including inflammatory DED and aqueous deficiency, ocular GVHD, or the like). In some embodiments, a compound provided herein is useful as either an acute therapy (e.g., by a trained specialist or physician) or as a chronic therapy (e.g., in the hands of a patient, or alternatively, by a trained specialist or physician). A compound provided herein is tested, in some embodiments, using the assays and methods described herein (e.g., as described in the examples). In some embodiments, a compound provided herein represents a significant advance in the art as the first-order metabolites obtained from metabolism of the agents are operative against both the keratolytic and the immunomodulation and/or inflammatory component of diseases, such as, for example, ocular allergies, dry eye disease, ocular GVHD, and the like.

[0307] Provided in some embodiments herein is a compound having a structure represented by Formula (A):

##STR00041##

wherein, [0308] X is an immunomodulator radical (e.g., an immunosuppressant radical or an immunostimulant radical); [0309] each G independently comprises at least one linker and at least one radical of a keratolytic agent; and [0310] n is 1-3,
or a pharmaceutically acceptable salt or solvate thereof.

[0311] Provided in some embodiments herein is a compound having a structure represented by Formula (A-I):

##STR00042##

wherein, [0312] X is an immunomodulator radical (e.g., an immunosuppressant radical or an immunostimulant radical); [0313] each Y.sup.1 and Y.sup.2 is independently a linker; [0314] each Z.sup.1 and Z.sup.2 is independently a radical of a keratolytic agent; and [0315] n is 1-3,
or a pharmaceutically acceptable salt or solvate thereof.

[0316] Provided in some embodiments herein is a compound having a structure represented by Formula (A-II):

##STR00043##

wherein, [0317] X is an immunomodulator radical (e.g., an immunosuppressant radical or an immunostimulant radical); [0318] each Y is independently a linker; [0319] each Z is independently a radical of a keratolytic agent; and [0320] n is 1-3, [0321] or a pharmaceutically acceptable salt or solvate thereof.

[0322] In some embodiments, X is an immunomodulator radical. In some embodiments, X is an immunosuppressant radical. In some embodiments, X is an immunostimulant radical.

[0323] In some embodiments, X (e.g., in its free form) is an anti-inflammatory agent.

[0324] In some embodiments, X (e.g., in its free form) is an immunomodulator (e.g., an immunosuppressant or an immunostimulant) and an anti-inflammatory agent.

[0325] In some embodiments, X is selected from the group consisting of a radical of a (e.g., selective) phosphodiesterase (PDE) inhibitor (e.g., a radical of a (e.g., selective) phosphodiesterase-4 (PDE-4) inhibitor (e.g., a cilomilast radical)), a radical of a (e.g., selective) Janus kinase (JAK) inhibitor (e.g., a radical of a JAK1, JAK2, and/or JAK3 inhibitor (e.g., a ruxolitinib radical, a tofacitinib radical, a oclacitinib radical, or a ritlecitinib radical), a radical of a folate reductase inhibitor (e.g., a methotrexate (or a derivative thereof) radical), a radical of a steroid (e.g., a corticosteroid radical (e.g., a glucocorticoid radical (e.g., a loteprednol (or a derivative thereof) radical))), and a radical of a calcineurin inhibitor (e.g., a tacrolimus radical).

[0326] In some embodiments, X is selected from the group consisting of a cilomilast radical, a ruxolitinib radical, a tofacitinib radical, an oclacitinib radical, a ritlecitinib radical, a methotrexate (or a derivative thereof) radical), a loteprednol (or a derivative thereof) radical, and a tacrolimus radical.

[0327] In some embodiments, X is selected from the group consisting of a cilomilast radical.

[0328] In some embodiments, X is selected from the group consisting of a ruxolitinib radical.

[0329] In some embodiments, X is selected from the group consisting of a tofacitinib radical.

[0330] In some embodiments, X is selected from the group consisting of an oclacitinib radical.

[0331] In some embodiments, X is selected from the group consisting of a ritlecitinib radical.

[0332] In some embodiments, X is selected from the group consisting of a methotrexate radical.

[0333] In some embodiments, X is selected from the group consisting of a loteprednol radical.

[0334] In some embodiments, X is selected from the group consisting of a tacrolimus radical.

[0335] In some embodiments, X is not lifitegrast.

[0336] In some embodiments, X is not azithromycin.

[0337] In some embodiments, X is not (S)-3-(4-((4-carbamoylpiperidine-1-carbonyl)oxy)phenyl)-2-((S)-4-methyl-2-(2-(o-tolyloxy)acetamido)pentanamido)propanoic acid.

[0338] In some embodiments, the anti-inflammatory agent is not lifitegrast. In some embodiments, the anti-inflammatory agent is not azithromycin. In some embodiments, the anti-inflammatory agent is not (S)-3-(4-((4-carbamoylpiperidine-1-carbonyl)oxy)phenyl)-2-((S)-4-methyl-2-(2-(o-tolyloxy)acetamido)pentanamido)propanoic acid.

[0339] In some embodiments, X has a structure represented by Formula (I-A):

##STR00044##

wherein, [0340] each R.sup.1 is independently halogen, alkyl, or CN; [0341] R.sup.2 is hydrogen, CN, halogen, or alkyl; [0342] each R.sup.3 is independently OR.sup.a, alkyl, heteroalkyl, cycloalkyl, or heterocyclyl; [0343] each R.sup.a is independently hydrogen, alkyl, cycloalkyl, or heterocyclyl; [0344] a is 0-9; and [0345] b is 0-5.

[0346] In some embodiments, a is 0. In some embodiments, R.sup.2 is CN. In some embodiments, b is 2. In some embodiments, each R.sup.3 is independently OR.sup.a. In some embodiments, each R.sup.a is independently alkyl or cycloalkyl. In some embodiments, each R.sup.3 is independently OMe or OC.sub.3-C.sub.5 cycloalkyl.

[0347] In some embodiments, X has a structure represented by Formula (I-AA):

##STR00045##

[0348] In some embodiments, X has a structure represented by Formula (I-B):

##STR00046##

wherein, [0349] R.sup.4 is hydrogen, halogen, or alkyl; [0350] R.sup.5 is hydrogen, halogen, or alkyl; [0351] R.sup.6 is hydrogen, halogen, or alkyl; [0352] R.sup.7 is NR.sup.bR.sup.c or optionally substituted heterocyclyl; and [0353] R.sup.b and R.sup.c are each independently hydrogen, alkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl.

[0354] In some embodiments, R.sup.4 and R.sup.5 are hydrogen. In some embodiments, R.sup.6 is hydrogen. In some embodiments, R.sup.7 is substituted heterocyclyl. In some embodiments, R.sup.7 is heterocyclyl substituted with alkyl substituted with CN and cycloalkyl.

[0355] In some embodiments, X has a structure represented by Formula (I-BA):

##STR00047##

[0356] In some embodiments, R.sup.4 and R.sup.5 are hydrogen. In some embodiments, R.sup.6 is hydrogen. In some embodiments, R.sup.7 is NR.sup.bR.sup.c. In some embodiments, R.sup.b is hydrogen or C.sub.1-C.sub.3 alkyl. In some embodiments, R.sup.c is substituted cycloalkyl or substituted heterocyclyl. In some embodiments, R.sup.b is hydrogen. In some embodiments, R.sup.c is heterocyclyl substituted with one or more substituent, each substituent being independently optionally substituted alkyl. In some embodiments, R.sup.c is heterocyclyl substituted with unsubstituted C.sub.1-C.sub.3 alkyl and (unsaturated) C.sub.1-C.sub.3 alkyl substituted with oxo.

[0357] In some embodiments, X has a structure represented by Formula (I-BB):

##STR00048##

[0358] In some embodiments, R.sup.4 and R.sup.5 are hydrogen. In some embodiments, R.sup.6 is hydrogen. In some embodiments, R.sup.7 is NR.sup.bR.sup.c. In some embodiments, R.sup.b is CH.sub.3. In some embodiments, R.sup.c is cycloalkyl or heterocyclyl substituted with one or more substituent, each substituent being independently optionally substituted alkyl. In some embodiments, R.sup.c is heterocyclyl substituted with unsubstituted C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkyl substituted with oxo and CN.

[0359] In some embodiments, X has a structure represented by Formula (I-BC):

##STR00049##

[0360] In some embodiments, R.sup.4 and R.sup.5 are hydrogen. In some embodiments, R.sup.6 is hydrogen. In some embodiments, R.sup.7 is NR.sup.bR.sup.c. In some embodiments, R.sup.b is CH.sub.3. In some embodiments, R.sup.c is cycloalkyl or heterocyclyl substituted with one or more substituent, each substituent being independently optionally substituted alkyl. In some embodiments, R.sup.c is cycloalkyl substituted with C.sub.1-C.sub.3 alkyl substituted with SO.sub.2NHCH.sub.3.

[0361] In some embodiments, X has a structure represented by Formula (I-BD):

##STR00050##

[0362] In some embodiments, X has a structure represented by Formula (I-C1), Formula (I-C2), or Formula (I-C3):

##STR00051##

wherein, [0363] R.sup.8 is hydrogen or alkyl; [0364] each R.sup.9 is independently halogen or alkyl; [0365] R.sup.10 is hydrogen or alkyl; [0366] R.sup.11 is a radical, hydrogen, or alkyl; [0367] R.sup.12 is a radical, hydrogen, or alkyl; [0368] R.sup.13 is a radical or hydrogen; [0369] d is 1-3; [0370] e is 0-4; and [0371] f is 1-4.

[0372] In some embodiments, R.sup.8 is CH.sub.3. In some embodiments, e is 0. In some embodiments, R.sup.10 is hydrogen. In some embodiments, d is 1. In some embodiments, f is 2. In some embodiments, R.sup.11 is a radical. In some embodiments, R.sup.12 is a radical. In some embodiments, R.sup.11 is a radical and R.sup.12 is a radical. In some embodiments, R.sup.13 is a radical. In some embodiments, R.sup.11 is a radical, R.sup.12 is a radical, and R.sup.13 is hydrogen.

[0373] In some embodiments, X has a structure represented by Formula (I-CA):

##STR00052##

[0374] In some embodiments, R.sup.8 is CH.sub.3. In some embodiments, e is 0. In some embodiments, R.sup.10 is hydrogen. In some embodiments, d is 1. In some embodiments, f is 2. In some embodiments, R.sup.13 is a radical and R.sup.11 and R.sup.12 are hydrogen.

[0375] In some embodiments, X has a structure represented by Formula (I-CB):

##STR00053##

[0376] In some embodiments, X has a structure represented by Formula (I-D):

##STR00054##

wherein, [0377] is a single bond or a double bond; [0378] R.sup.14 is hydrogen, or optionally substituted alkyl; [0379] R.sup.21 is hydrogen, halogen, or alkyl; [0380] R.sup.22 is hydrogen, halogen, or alkyl; [0381] R.sup.23 is hydrogen or alkyl; [0382] R.sup.24 is hydrogen or alkyl; and [0383] R.sup.25 is hydrogen or alkyl.

[0384] In some embodiments, is a single bond. In some embodiments, is a double bond. In some embodiments, R.sup.14 is hydrogen or optionally substituted alkyl. In some embodiments, R.sup.14 is hydrogen. In some embodiments, R.sup.14 is optionally substituted alkyl. In some embodiments, R.sup.14 is alkyl substituted with halo or cyano. In some embodiments, R.sup.14 is alkyl substituted with halo. In some embodiments, R.sup.14 is alkyl substituted with chloro. In some embodiments, R.sup.14 is alkyl substituted with cyano. In some embodiments, R.sup.21 is hydrogen, halogen, or alkyl. In some embodiments, R.sup.21 is hydrogen. In some embodiments, R.sup.21 is halogen. In some embodiments, R.sup.21 is alkyl. In some embodiments, R.sup.21 is hydrogen or halogen. In some embodiments, R.sup.21 is fluorine. In some embodiments, R.sup.22 is hydrogen, halogen, or alkyl. In some embodiments, R.sup.22 is hydrogen. In some embodiments, R.sup.22 is halogen. In some embodiments, R.sup.22 is alkyl. In some embodiments, R.sup.22 is hydrogen or halogen. In some embodiments, R.sup.22 is fluorine. In some embodiments, R.sup.23 is hydrogen or alkyl. In some embodiments, R.sup.23 is hydrogen. In some embodiments, R.sup.23 is alkyl. In some embodiments, R.sup.23 is methyl. In some embodiments, R.sup.24 is hydrogen or alkyl. In some embodiments, R.sup.24 is hydrogen. In some embodiments, R.sup.24 is alkyl. In some embodiments, R.sup.24 is methyl. In some embodiments, R.sup.25 is hydrogen or alkyl. In some embodiments, R.sup.25 is hydrogen. In some embodiments, R.sup.25 is alkyl. In some embodiments, R.sup.25 is methyl. In some embodiments, R.sup.24 and R.sup.25 are methyl. In some embodiments, R.sup.23, R.sup.24, and R.sup.25 are methyl. In some embodiments, R.sup.21 and R.sup.22 are fluorine.

[0385] In some embodiments, X has a structure represented by Formula (I-D):

##STR00055##

wherein, [0386] is a single bond or a double bond; and [0387] R.sup.14 is hydrogen or optionally substituted alkyl.

[0388] In some embodiments, is a double bond. In some embodiments, R.sup.14 is alkyl substituted with halogen. In some embodiments, R.sup.14 is hydrogen or alkyl substituted with halogen or cyano. In some embodiments, R.sup.14 is hydrogen. In some embodiments R.sup.14 is alkyl substituted with halo. In some embodiments R.sup.14 is alkyl substituted with chloro.

[0389] In some embodiments, X has a structure represented by Formula (I-DA):

##STR00056##

[0390] In some embodiments, is a double bond. In some embodiments, R.sup.14 is hydrogen.

[0391] In some embodiments, X has a structure represented by Formula (I-DB):

##STR00057##

[0392] In some embodiments, R.sup.14 is alkyl substituted with cyano.

[0393] In some embodiments, X has a structure represented by Formula (I-DC):

##STR00058##

[0394] In some embodiments, X has a structure represented by Formula (I-E) or Formula (I-E):

##STR00059## [0395] wherein, [0396] R.sup.15, R.sup.16, and R.sup.17 are each independently selected from a radical, hydrogen, or alkyl.

[0397] In some embodiments, R.sup.15 is a radical. In some embodiments, R.sup.16 is a radical. In some embodiments, R.sup.15 is a radical and R.sup.16 is a radical. In some embodiments, R.sup.17 is hydrogen.

[0398] In some embodiments, R.sup.15 is a radical, R.sup.16 is hydrogen, and R.sup.17 is hydrogen.

[0399] In some embodiments, X has a structure represented by Formula (I-EA):

##STR00060##

[0400] In some embodiments, R.sup.15 is hydrogen, R.sup.16 is a radical, and R.sup.17 is hydrogen.

[0401] In some embodiments, X has a structure represented by Formula (I-EB):

##STR00061##

[0402] In some embodiments, R.sup.15 is a radical, R.sup.16 is a radical, and R.sup.17 is hydrogen.

[0403] In some embodiments, X has a structure represented by Formula (I-EC):

##STR00062##

[0404] In some embodiments, n is 1. In some embodiments, n is 2.

[0405] In some embodiments, each linker (e.g., Y, Y.sup.1, or Y.sup.2) is the same.

[0406] In some embodiments, each linker (e.g., Y, Y.sup.1, or Y.sup.2) is different.

[0407] In some embodiments, each linker (e.g., Y, Y.sup.1, or Y.sup.2) is independently a bond, substituted or unsubstituted (e.g., branched or straight) alkyl (alkylenyl), or substituted or unsubstituted (e.g., branched or straight) heteroalkyl (heteroalkylenyl). In some embodiments, each linker (e.g., Y, Y.sup.1, or Y.sup.2) is independently substituted or unsubstituted (e.g., branched or straight) alkyl (alkylenyl) or substituted or unsubstituted (e.g., branched or straight) heteroalkyl (heteroalkylenyl). In some embodiments, each linker (e.g., Y, Y.sup.1, or Y.sup.2) is independently a bond, CH(CH.sub.3), or CH.sub.2. In some embodiments, each linker (e.g., Y, Y.sup.1, or Y.sup.2) is a bond. In some embodiments, Y.sup.1 is a bond and each Y.sup.2 is independently CH(CH.sub.3) or CH.sub.2.

[0408] In some embodiments, each radical of a keratolytic agent (e.g., Z, Z.sup.1, or Z.sup.2) is a radical of the same keratolytic agent.

[0409] In some embodiments, each radical of a keratolytic agent (e.g., Z, Z.sup.1, or Z.sup.2) is a radical of a different keratolytic agent.

[0410] In some embodiments, each radical of a keratolytic agent (e.g., Z, Z.sup.1, or Z.sup.2) comprises one or more keratolytic group. In some embodiments, each radical of a keratolytic agent (e.g., Z, Z.sup.1, or Z.sup.2) comprises one or more group, each group being independently selected from the group consisting of O, oxo, substituted or unsubstituted (e.g., branched or straight) alkyl (alkylenyl), substituted or unsubstituted (e.g., branched or straight) heteroalkyl (heteroalkylenyl), substituted or unsubstituted alkoxyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocyclyl. In some embodiments, each radical of a keratolytic agent (e.g., Z, Z.sup.1, or Z.sup.2) is (e.g., branched or straight) alkyl (alkylenyl) substituted with one or more substituent, each substituent being independently selected from the group consisting of oxo, hydroxy, alkyl, alkoxy, and substituted or unsubstituted heterocyclyl. In some embodiments, each radical of a keratolytic agent (e.g., Z, Z.sup.1, or Z.sup.2) is (e.g., branched or straight) heteroalkyl (heteroalkylenyl) substituted with one or more substituent, each substituent being independently selected from the group consisting of oxo, alkyl, thioalkyl, and substituted or unsubstituted heterocyclyl. In some embodiments, Z.sup.1 is straight alkyl (alkylenyl) substituted with one or more substituent, each substituent being independently selected from the group consisting of oxo and hydroxy and Z.sup.2 is straight alkyl (alkylenyl) substituted with one or more substituent, each substituent being independently selected from the group consisting of oxo and substituted or unsubstituted heterocyclyl.

[0411] In some embodiments, Y.sup.1 is a bond, Y.sup.2 is CH(CH.sub.3) or CH.sub.2, Z.sup.1 is

##STR00063##

n is 1.

[0412] In some embodiments, each radical of a keratolytic agent (e.g., Z, Z.sup.1, or Z.sup.2) has a structure represented by:

##STR00064##

wherein: [0413] Q is O or (CR.sup.18R.sup.19).sub.m; [0414] m is 1-6; [0415] each R.sup.18 and R.sup.19 is independently H, halo, alkyl, alkoxy, haloalkyl, or thioalkyl; [0416] or an adjacent R.sup.18 and R.sup.19 combine to the atoms to which they are attached to form an oxo; and [0417] R.sup.20 is alkyl, heteroalkyl, heterocyclyl, alkoxy, or hydroxy, the alkyl, heteroalkyl, heterocyclyl, or alkoxy each independently being optionally substituted.

[0418] In some embodiments, each R.sup.18 and R.sup.19 is independently H, C.sub.1-C.sub.6 alkyl, or C.sub.1-C.sub.3 thioalkyl. In some embodiments, each R.sup.18 and R.sup.19 is independently H, CH.sub.3, or CH.sub.2SH. In some embodiments, m is 1-4.

[0419] In some embodiments, Q is (CR.sup.18R.sup.19).sub.m, m is 1-4, R.sup.18 and R.sup.19 are each independently H or C.sub.1-C.sub.6 alkyl, and R.sup.20 is optionally substituted heterocyclyl.

[0420] In some embodiments, R.sup.20 is dithiolanyl or dithiolanyl oxide. In some embodiments, R.sup.20 is:

##STR00065##

[0421] In some embodiments, Q is CH.sub.2, CH(CH.sub.3), (CH.sub.2).sub.2C(O), CH.sub.2C(CH.sub.3).sub.2CH.sub.2, or CH(CH.sub.2SH) and R.sup.20 is hydroxy, optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.1-C.sub.6 alkoxy, or optionally substituted C.sub.1-C.sub.6 heteroalkyl.

[0422] In some embodiments, R.sup.20 is CH.sub.3, hydroxy, O(C.sub.1-C.sub.3 alkoxy), or substituted C.sub.1-C.sub.6 heteroalkyl (e.g., heteroalkyl substituted with CH.sub.3, oxo, and dithiolanyl or dithiolanyl oxide). In some embodiments, R.sup.20 is OH, CH.sub.3, OCH.sub.3, OCH.sub.2CH.sub.3, NH(CO)CH.sub.3,

##STR00066##

[0423] In some embodiments, Q is O and R.sup.20 is optionally substituted C.sub.1-C.sub.6 alkyl.

[0424] In some embodiments, R.sup.20 is methyl, ethyl, propyl, isopropyl, butyl, or tert-butyl.

[0425] Provided in some embodiments herein is a compound having a structure provided in Table 1, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00001 TABLE 1 [00067] Com- pound L.sup.1 Q.sup.1 1 CH(CH.sub.3) [00068] 2 CH(CH.sub.3) [00069] 3 CH(CH.sub.3) [00070] 4 CH.sub.2 [00071] 5 CH.sub.2 [00072] 6 CH.sub.2 [00073]

[0426] Provided in some embodiments herein is a compound having a structure provided in Table 2, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00002 TABLE 2 [00074] Compound L.sup.2 Q.sup.2 7 CH(CH.sub.3) [00075] 8 bond [00076] 9 CH.sub.2 [00077] 10 CH(CH.sub.3) [00078] 11 CH.sub.2 [00079] 12 bond [00080] 13 bond [00081] 14 bond [00082] 15 bond [00083] 16 bond [00084]

[0427] Provided in some embodiments herein is a compound having a structure provided in Table 3, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00003 TABLE 3 [00085] Compound L.sup.3 Q.sup.3 17 bond [00086] 18 bond [00087] 19 CH(CH.sub.3) [00088] 20 bond [00089] 21 CH.sub.2 [00090] 22 bond [00091] 23 bond [00092] 24 bond [00093]

[0428] Provided in some embodiments herein is a compound having a structure provided in Table 4, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00004 TABLE 4 [00094] Com- pound L.sup.4 Q.sup.4 25 CH(CH.sub.3) [00095] 26 CH(CH.sub.3) [00096] 27 CH.sub.2 [00097] 28 CH.sub.2 [00098] 29 CH(CH.sub.3) [00099] 30 bond [00100] 31 bond [00101] 32 bond [00102] 33 bond [00103]

[0429] Provided in some embodiments herein is a compound having a structure provided in Table 5, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00005 TABLE 5 [00104] Compound L.sup.5 Q.sup.5 34 CH(CH.sub.3) [00105] 35 CH(CH.sub.3) [00106] 36 bond [00107] 37 CH.sub.2 [00108] 38 CH.sub.2 [00109] 39 bond [00110] 40 bond [00111] 41 CH(CH.sub.3) [00112] 42 CH(CH.sub.3) [00113] 43 bond [00114] 44 bond [00115] 45 bond [00116] 46 bond [00117]

[0430] Provided in some embodiments herein is a compound having a structure provided in Table 6, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00006 TABLE 6 [00118] Compound L.sup.6a L.sup.6b Q.sup.6a Q.sup.6b 47 CH(CH.sub.3) CH(CH.sub.3) [00119] [00120] 48 CH.sub.2 CH.sub.2 [00121] [00122] 49 CH(CH.sub.3) CH.sub.2 [00123] [00124] 50 CH.sub.2 CH(CH.sub.3) [00125] [00126] 51 CH(CH.sub.3) bond [00127] CH.sub.2CH.sub.3 52 CH.sub.2 bond [00128] CH.sub.2CH.sub.3 53 bond CH(CH.sub.3) CH.sub.2CH.sub.3 [00129] 54 bond CH.sub.2 CH.sub.2CH.sub.3 [00130] 55 CH(CH.sub.3) CH(CH.sub.3) [00131] [00132] 56 bond bond CH(CH.sub.3).sub.2 CH(CH.sub.3).sub.2 57 CH(CH.sub.3) CH(CH.sub.3) [00133] [00134] 58 bond bond CH.sub.2CH.sub.3 CH.sub.2CH.sub.3 59 bond bond CH.sub.2CH.sub.3 H 60 bond bond H CH.sub.2CH.sub.3

[0431] Provided in some embodiments herein is a compound having a structure provided in Table 7, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00007 TABLE 7 [00135] Compound L.sup.7 Q.sup.7 61 CH(CH.sub.3) [00136] 62 CH.sub.2 [00137] 63 bond [00138] 64 bond [00139]

[0432] Provided in some embodiments herein is a compound having a structure provided in Table 8, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00008 TABLE 8 [00140] Compound Q.sup.x L.sup.8 Q.sup.8 65 ClCH.sub.2 bond [00141] 66 ClCH.sub.2 bond [00142] 67 ClCH.sub.2 bond [00143] 68 ClCH.sub.2 bond [00144] 69 H bond [00145] 70 H bond [00146] 71 ClCH.sub.2 bond [00147] 72 H bond [00148] 73 ClCH.sub.2 bond H

[0433] Provided in some embodiments herein is a compound having a structure provided in Table 8A, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00009 TABLE 8A [00149] Compound Q.sup.y L.sup.8A Q.sup.8A 73A NC(CH).sub.2 bond [00150] 73B NC(CH).sub.2 bond [00151]

[0434] In some instances, a free-acid is inactive in the glucocorticoid binding assay described in the Examples herein. In some instances, an ester is active in the glucocorticoid binding assay described in the Examples herein. In some instances, an ester retains activity, such as when attached to a keratolytic agent (or a radical thereof), in the glucocorticoid binding assay described in the Examples herein.

[0435] Provided in some embodiments herein is a compound having a structure provided in Table 9, a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate of the compound or the stereoisomer.

TABLE-US-00010 TABLE 9 [00152] Compound L.sup.9a L.sup.9b Q.sup.9a Q.sup.9b 74 bond bond [00153] H 75 bond bond [00154] H 76 bond bond [00155] H 77 bond bond H [00156] 78 bond bond H [00157] 79 bond bond H [00158] 80 bond bond [00159] [00160] 81 bond bond [00161] [00162] 82 bond bond [00163] [00164]

[0436] Each recitation of

##STR00165##

provided herein, unless otherwise stated, includes a specific and explicit recitation of:

##STR00166##

[0437] Each recitation of

##STR00167##

provided herein, unless otherwise stated, includes a specific

##STR00168##

[0438] The compounds used in the reactions described herein are made according to organic synthesis techniques starting from commercially available chemicals and/or from compounds described in the chemical literature or provided herein. Commercially available chemicals are obtained from standard commercial sources including Acros Organics (Pittsburgh, PA), Aldrich Chemical (Milwaukee, WI, including Sigma Chemical and Fluka), Apin Chemicals Ltd. (Milton Park, UK), Avocado Research (Lancashire, U.K.), BDH Inc. (Toronto, Canada), Bionet (Cornwall, U.K.), Chemservice Inc. (West Chester, PA), Crescent Chemical Co. (Hauppauge, NY), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scientific Co. (Pittsburgh, PA), Fisons Chemicals (Leicestershire, UK), Frontier Scientific (Logan, UT), ICN Biomedicals, Inc. (Costa Mesa, CA), Key Organics (Cornwall, U.K.), Lancaster Synthesis (Windham, NH), Maybridge Chemical Co. Ltd. (Cornwall, U.K.), Parish Chemical Co. (Orem, UT), Pfaltz & Bauer, Inc. (Waterbury, CN), Polyorganix (Houston, TX), Pierce Chemical Co. (Rockford, IL), Riedel de Haen A G (Hanover, Germany), Spectrum Quality Product, Inc. (New Brunswick, NJ), TCI America (Portland, OR), Trans World Chemicals, Inc. (Rockville, MD), and Wako Chemicals USA, Inc. (Richmond, VA).

[0439] Suitable reference books and treatise that detail the synthesis of reactants useful in the preparation of compounds described herein, or provide references to articles that describe the preparation, include for example, Synthetic Organic Chemistry, John Wiley & Sons, Inc., New York; S. R. Sandler et al., Organic Functional Group Preparations, 2.sup.nd Ed., Academic Press, New York, 1983; H. 0. House, Modern Synthetic Reactions, 2.sup.nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif 1972; T. L. Gilchrist, Heterocyclic Chemistry, 2.sup.nd Ed., John Wiley & Sons, New York, 1992; J. March, Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 4.sup.th Ed., Wiley-Interscience, New York, 1992. Additional suitable reference books and treatise that detail the synthesis of reactants useful in the preparation of compounds described herein, or provide references to articles that describe the preparation, include for example, Fuhrhop, J. and Penzlin G. Organic Synthesis: Concepts, Methods, Starting Materials, Second, Revised and Enlarged Edition (1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R.V. Organic Chemistry, An Intermediate Text (1996) Oxford University Press, ISBN 0-19-509618-5; Larock, R. C. Comprehensive Organic Transformations: A Guide to Functional Group Preparations 2.sup.nd Edition (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. Advanced Organic Chemistry: Reactions, Mechanisms, and Structure 4.sup.th Edition (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Otera, J. (editor) Modern Carbonyl Chemistry (2000) Wiley-VCH, ISBN: 3-527-2987 1-1; Patai, S. Patai's 1992 Guide to the Chemistry of Functional Groups (1992) Interscience ISBN: 0-471-93022-9; Solomons, T. W. G. Organic Chemistry 7.sup.th Edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0; Stowell, J. C., Intermediate Organic Chemistry 2.sup.nd Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; Industrial Organic Chemicals: Starting Materials and Intermediates: An Ullmann's Encyclopedia (1999) John Wiley & Sons, ISBN: 3-527-29645-X, in 8 volumes; Organic Reactions (1942-2000) John Wiley & Sons, in over 55 volumes; and Chemistry of Functional Groups John Wiley & Sons, in 73 volumes.

[0440] Specific and analogous reactants are optionally identified through the indices of chemicals prepared by the Chemical Abstract Service of the American Chemical Society, which are available in most public and university libraries, as well as through on-line databases (contact the American Chemical Society, Washington, D.C. for more details). Chemicals not commercially available in catalogs are optionally prepared by custom chemical synthesis houses, where many of the standard chemical supply houses (e.g., those listed above) provide custom synthesis services. A reference for the preparation and selection of pharmaceutical salts of the keratolytic conjugate described herein is P. H. Stahl & C. G. Wermuth Handbook of Pharmaceutical Salts, Verlag Helvetica Chimica Acta, Zurich, 2002.

[0441] In some embodiments, a compound provided herein is represented by any one of Formula (A), Formula (A-I), Formula (A-II), Formula (I-A), Formula (I-AA), Formula (I-B), Formula (I-BA), Formula (I-BB), Formula (I-BC), Formula (I-BD), Formula (I-C1), Formula (I-C2), Formula (I-C3), Formula (I-CA), Formula (I-CB), Formula (I-D), Formula (I-DA), Formula (I-DB), Formula (I-E), Formula (I-E), Formula (I-EA), Formula (I-EB), Formula (I-EC), Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9. In some embodiments, a compound provided herein is administered as a pure chemical. In other embodiments, a compound provided herein is combined with a pharmaceutically suitable or acceptable carrier (also referred to herein as a pharmaceutically suitable (or acceptable) excipient, physiologically suitable (or acceptable) excipient, or physiologically suitable (or acceptable) carrier) selected on the basis of a chosen route of administration and standard pharmaceutical practice as described, for example, in Remington: The Science and Practice of Pharmacy (Gennaro, 21.sup.st Ed. Mack Pub. Co., Easton, PA (2005)).

[0442] Provided in some embodiments herein is a pharmaceutical composition comprising at least one keratolytic conjugate together with one or more pharmaceutically acceptable carriers. The carrier(s) (or excipient(s)) is acceptable or suitable if the carrier is compatible with the other ingredients of the composition and not deleterious to the recipient (i.e., the subject) of the composition.

[0443] In some embodiments, a compound provided herein (e.g., a compound having a structure represented by any one of Formula (A), Formula (A-I), Formula (A-II), Formula (I-A), Formula (I-AA), Formula (I-B), Formula (I-BA), Formula (I-BB), Formula (I-BC), Formula (I-BD), Formula (I-C1), Formula (I-C2), Formula (I-C3), Formula (I-CA), Formula (I-CB), Formula (I-D), Formula (I-DA), Formula (I-DB), Formula (I-E), Formula (I-E), Formula (I-EA), Formula (I-EB), Formula (I-EC), Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9) is substantially pure, in that it contains less than, for example, about 5%, or less than about 1%, or less than about 0.1%, of other organic small molecules, such as unreacted intermediates or synthesis by-products that are created, for example, in one or more of the steps of a synthesis method.

[0444] Suitable oral dosage forms include, for example, tablets, pills, sachets, or capsules of hard or soft gelatin, methylcellulose or of another suitable material easily dissolved in the digestive tract. In some embodiments, suitable nontoxic solid carriers are used which include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and the like. (See, e.g., Remington: The Science and Practice of Pharmacy (Gennaro, 21.sup.st Ed. Mack Pub. Co., Easton, PA (2005)).

[0445] In some embodiments provided herein is a pharmaceutical composition comprising a compound provided herein (e.g., a compound having a structure represented by any one of Formula (A), Formula (A-I), Formula (A-II), Formula (I-A), Formula (I-AA), Formula (I-B), Formula (I-BA), Formula (I-BB), Formula (I-BC), Formula (I-BD), Formula (I-C1), Formula (I-C2), Formula (I-C3), Formula (I-CA), Formula (I-CB), Formula (I-D), Formula (I-DA), Formula (I-DB), Formula (I-E), Formula (I-E), Formula (I-EA), Formula (I-EB), Formula (I-EC), Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9) and at least one pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is suitable for dermal administration. In some embodiments, the pharmaceutical composition is suitable for ophthalmic administration. In some embodiments, the pharmaceutical composition is suitable for topical ophthalmic administration. In some embodiments, topical ophthalmic administration is administration in and/or around the eye, such as to the eyelid margin. In some embodiments, the pharmaceutical composition is suitable for administration to the eyelid margin. In some embodiments, topical ophthalmic administration is administration to the ocular surface and the inner surface to the eyelid.

[0446] In some embodiments, a keratolytic conjugate provided herein (e.g., a compound having a structure represented any one of Formula (A), Formula (A-I), Formula (A-II), Formula (I-A), Formula (I-AA), Formula (I-B), Formula (I-BA), Formula (I-BB), Formula (I-BC), Formula (I-BD), Formula (I-C1), Formula (I-C2), Formula (I-C3), Formula (I-CA), Formula (I-CB), Formula (I-D), Formula (I-DA), Formula (I-DB), Formula (I-E), Formula (I-E), Formula (I-EA), Formula (I-EB), Formula (I-EC), Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9) is formulated as a solution or suspension for topical administration to the eye.

[0447] In some embodiments, a keratolytic conjugate provided herein (e.g., a compound having a structure represented any one of Formula (A), Formula (A-I), Formula (A-II), Formula (I-A), Formula (I-AA), Formula (I-B), Formula (I-BA), Formula (I-BB), Formula (I-BC), Formula (I-BD), Formula (I-C1), Formula (I-C2), Formula (I-C3), Formula (I-CA), Formula (I-CB), Formula (I-D), Formula (I-DA), Formula (I-DB), Formula (I-E), Formula (I-E), Formula (I-EA), Formula (I-EB), Formula (I-EC), Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9) is formulated as a solution or suspension for topical administration to the skin.

[0448] In some embodiments, a keratolytic conjugate provided herein (e.g., a compound having a structure represented by any one of Formula (A), Formula (A-I), Formula (A-II), Formula (I-A), Formula (I-AA), Formula (I-B), Formula (I-BA), Formula (I-BB), Formula (I-BC), Formula (I-BD), Formula (I-C1), Formula (I-C2), Formula (I-C3), Formula (I-CA), Formula (I-CB), Formula (I-D), Formula (I-DA), Formula (I-DB), Formula (I-E), Formula (I-E), Formula (I-EA), Formula (I-EB), Formula (I-EC), Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9) is formulated for administration by injection. In some instances, the injection formulation is an aqueous formulation. In some instances, the injection formulation is a non-aqueous formulation. In some instances, the injection formulation is an oil-based formulation, such as sesame oil, or the like.

[0449] In some embodiments, the dose of the composition comprising at least one keratolytic conjugate as provided herein differ, depending upon the patient's (e.g., human) condition, that is, general health status, age, and other factors.

[0450] Pharmaceutical compositions provided in some embodiments herein are administered in a manner appropriate to the disease to be treated (or prevented). An appropriate dose and a suitable duration and frequency of administration will be determined by such factors as the condition of the patient, the type and severity of the patient's disease, the particular form of the active ingredient, and the method of administration. In general, an appropriate dose and treatment regimen provides the composition(s) in an amount sufficient to provide therapeutic and/or prophylactic benefit (e.g., an improved clinical outcome, such as more frequent complete or partial remissions, or longer disease-free and/or overall survival, or a lessening of symptom severity). Optimal doses are generally determined using experimental models and/or clinical trials. The optimal dose depends upon the body mass, weight, or blood volume of the patient.

[0451] In other embodiments, the topical compositions described herein are combined with a pharmaceutically suitable or acceptable carrier (e.g., a pharmaceutically suitable (or acceptable) excipient, physiologically suitable (or acceptable) excipient, or physiologically suitable (or acceptable) carrier). Exemplary excipients are described, for example, in Remington: The Science and Practice of Pharmacy (Gennaro, 21st Ed. Mack Pub. Co., Easton, PA (2005)).

Methods of Treatment Utilizing Keratolytic Conjugates

[0452] In some embodiments provided herein is a method of treating a dermatological or ophthalmic disease or disorder in a patient in need of thereof, comprising administering to the patient any compound provided herein, or a pharmaceutically acceptable salt thereof, or a (e.g., pharmaceutical) composition comprising any compound provided herein, or a pharmaceutically acceptable salt thereof, such as a compound represented by any structure herein, such as, for example, Formula (A), Formula (A-I), Formula (A-II), Formula (I-A), Formula (I-AA), Formula (I-B), Formula (I-BA), Formula (I-BB), Formula (I-BC), Formula (I-BD), Formula (I-C1), Formula (I-C2), Formula (I-C3), Formula (I-CA), Formula (I-CB), Formula (I-D), Formula (I-DA), Formula (I-DB), Formula (I-E), Formula (I-E), Formula (I-EA), Formula (I-EB), Formula (I-EC), Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9. In some embodiments provided herein the pharmaceutical composition is in the form of a solution or suspension suitable for (e.g., topical) ophthalmic administration. In some embodiments, (e.g., topical) ophthalmic administration is administration in and/or around the eye, such as to the eyelid margin. In some embodiments, (e.g., topical) ophthalmic administration is administration to the ocular surface and the inner surface to the eyelid. In some embodiments, the dermatological or ophthalmic disease or disorder is improper immunomodulation, inflammation, and/or hyperkeratosis (e.g., of the eyes or skin). In some embodiments, the dermatological or ophthalmic disease or disorder is improper immunomodulation, inflammation or hyperkeratosis of the eyes or skin (e.g., the ocular surface). In some embodiments, the dermatological or ophthalmic dermatological disease or disorder is selected from the group consisting of meibomian gland dysfunction (MGD), dry eye disease (DED), ocular manifestations of graft versus host disease, vernal keratoconjunctivitis, atopic keratoconjunctivitis, Cornelia de Lange Syndrome, evaporative eye disease, aqueous deficiency dry eye, blepharitis, and seborrheic blepharitis. In some embodiments, the dermatological or ophthalmic disease or disorder is inflammation or hyperkeratosis (e.g., of the eyes or skin), such as, for example, meibomian gland dysfunction (MGD), dry eye disease (DED), ocular manifestations of graft versus host disease, vernal keratoconjunctivitis, atopic keratoconjunctivitis, Cornelia de Lange Syndrome, evaporative eye disease, aqueous deficiency dry eye, blepharitis, seborrheic blepharitis, or any combination thereof.

[0453] In some embodiments, the ophthalmic disease or disorder is selected from the group consisting of dry eye, lid wiper epitheliopathy (LWE), contact lens discomfort (CLD), dry eye syndrome, evaporative dry eye syndrome, aqueous deficiency dry eye syndrome, blepharitis, keratitis, meibomian gland dysfunction, conjunctivitis, lacrimal gland disorder, contact lens related conditions and inflammation of the anterior surface of the eye, infection of the anterior surface of the eye, and autoimmune disorder of the anterior surface of the eye.

[0454] In certain embodiments, methods provided herein involve the method of treating meibomian gland dysfunction (MGD).

[0455] In some embodiments, the ocular disorder is a surface disorder, such as MGD, dry eye and associated inflammatory and bacterial disease, an (e.g., severe) ocular allergy (e.g., keratoconjunctivitis (e.g., atopic keratoconjunctivitis (AKC) or vernal keratoconjunctivitis (VKC))), a (e.g., inflammatory and/or aqueous) dry eye disease, or an ocular manifestation of graft versus host disease (ocular GVHD).

[0456] In some embodiments, the ocular disorder is a periocular disorder. In some embodiments, the periocular disorder is a sty, blepharitis, a chalazion, or dacryoadenitis.

[0457] In some embodiments, the dermal disorder is comedonal acne, hyperkeratosis, scleroderma, seborrheic dermatitis, atopic dermatitis, psoriasis, lichen planus, an insect bite, intertrigo, pemphigus, or pityriasis rubra pilaris.

[0458] Provided herein is a method for treating an ocular surface disorder in an individual in need thereof comprising topical administration of a keratolytic conjugate to the individual in need thereof. In some embodiments, administration occurs with the assistance of a health-care provider (e.g., this category includes both acute and maintenance uses of the keratolytic conjugate). An acute use, in some embodiments, requires a stronger keratolytic conjugate (either in terms of concentration of the agent or the inherent activity of the agent). A maintenance use, in some embodiments, allows for the use of lower concentrations of the agent, or agents with lower inherent activity. A maintenance use, in some embodiments, involves a patient at a routine visit to the health care provider. Both acute uses and maintenance uses optionally involve use of an eye-protecting device or apparatus. In some embodiments, the acute use is performed by the health care provider, and the maintenance use is performed by the patient or non-health care provider. In some embodiments, administration does not occur with the active assistance of a health care provider (e.g., but rather involves the patient applying the keratolytic conjugate to his/her own eyelid margin). In some embodiments, such administration occurs over an extended period of time (e.g., one way of describing this patient-administered multi-administration mode is as a chronic use). In some embodiments, different or second formulations of the keratolytic conjugate are used for chronic or patient-administered uses. In some embodiments the different or second formulation utilizes a lower concentration of the keratolytic conjugate. In some embodiments, the second or different formulation utilizes a keratolytic conjugate that has a lower activity than the first formulation.

[0459] It should be understood that the present methods also include the physical removal of an obstruction in an meibomian gland (e.g., followed by chronic and/or maintenance administration of a keratolytic conjugate provided herein).

[0460] In some embodiments provided herein is a method for treating meibomian gland dysfunction in a patient in need thereof, comprising topically administering to the patient a composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier. In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier results in enhanced meibum production.

[0461] In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier occurs until the keratinized obstruction is relieved. In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier occurs periodically after relieving of the keratinized obstruction. In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is a single administration. In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is a periodic administration. In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier occurs once a day. In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier occurs twice a day. In some embodiments, the topical administration of the composition comprising a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier occurs more than twice a day.

[0462] In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is a solution. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is a solution suitable for topical administration as eye drops. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is a gel, ocular insert, spray, or other topical ocular delivery method. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is a semi-solid. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is homogenous. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is a dispersion. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier is hydrophilic. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier and an oleaginous base. In some embodiments, the composition for topical administration comprises a therapeutically-effective amount of at least one keratolytic conjugate in an ophthalmically-acceptable carrier and at least one ophthalmically-acceptable excipient.

[0463] In some embodiments provided herein is a method for treating MGD in a patient in need thereof comprising topical administration of a composition comprising a keratolytic conjugate. In some embodiments, the topical administration of the composition comprising a keratolytic conjugate occurs once a week. In some embodiments, the topical administration of the composition comprising a keratolytic conjugate occurs twice a week. In some embodiments, the topical administration of the composition comprising a keratolytic conjugate occurs every other day. In some embodiments, the topical administration of the composition comprises a keratolytic conjugate occurs every day. In some embodiments, the topical administration of the composition comprises a keratolytic conjugate occurs several times a day.

[0464] In some embodiments, the method comprises administering a compound or formulation provided herein in an acute treatment scenario. In some embodiments, the method comprises treatment of a patient naive to treatment. In some embodiments, the method comprises administering a compound or formulation provided herein in a chronic treatment scenario. In some embodiments, the method comprises administering a compound or formulation provided herein in a maintenance therapy scenario. In an acute treatment scenario, the administered dosage of keratolytic conjugate may be higher than the administered dosage of keratolytic conjugate employed in a chronic treatment scenario or a maintenance therapy scenario. In an acute treatment scenario, the keratolytic conjugate may be different from the keratolytic conjugate employed in a chronic treatment scenario. In some embodiments, the course of therapy begins in the initial phase of therapy as an acute treatment scenario and later transitions into a chronic treatment scenario or a maintenance therapy scenario. In some embodiments, the meibomian gland opening pharmacological agent administered in the acute treatment scenario is a keratolytic agent and/or keratoplastic agent, and the pharmacological agent administered in the chronic treatment scenario or a maintenance therapy scenario is a keratolytic conjugate.

[0465] In some embodiments, an initial treatment is administered (e.g., by a physician or healthcare professional) to an individual to initially open a blockage of the meibomian gland, such as by placing a more highly concentrated formulation of one of the keratolytic conjugate provided herein. In the event the higher concentration formulations are required, the application thereof may require ocular shielding or other activity to minimize the impact of irritation or disruption of the ocular surface or surrounding tissues. Following such a procedure, a patient may be given a different formulation of keratolytic conjugate to take home to apply periodically to the lid margin to maintain the patency of the meibomian gland. Such application may occur twice daily, once a day, weekly or monthly, depending on the formulation activity and the therapeutic product profile of the formulation.

[0466] Provided in some embodiments of the methods of treatment described herein is the location of the topical administration of the composition. In some embodiments, the composition comprising a keratolytic conjugate is administered such that no irritation to eye occurs. In some embodiments, the composition comprising a keratolytic conjugate is administered to the eye lid margin.

[0467] In some embodiments of the methods of treatment provided herein is the use of a protective element provided to the eye to avoid irritation to the eye. Although the formulations described herein are generally non-irritating, in some embodiments (e.g., high concentration of agent or when used on a sensitive eye) a protective element provides an additional layer of safety and comfort for the patient. In some embodiments, the composition comprising a keratolytic conjugate is administered while an eye shield is placed on the eye to reduce contact of the pharmacological agent with the cornea and/or conjunctiva such that reduced irritation to eye occurs. In some embodiments, the eye shield is a contact lens or an eye covering. In some embodiments, the eye covering comprises a self-adhesive. In some embodiments, the composition comprising a keratolytic conjugate is administered while the lid is pulled away from the globe to reduce contact of the pharmacological agent with the cornea and/or conjunctiva such that reduced irritation to eye occurs.

[0468] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

EXAMPLES

I. Chemical Synthesis

[0469] Solvents, reagents, and starting materials were purchased from commercial vendors and used as received unless otherwise described. All reactions were performed at room temperature unless otherwise stated. Starting materials were purchased from commercial sources or synthesized according to the methods described herein or using literature procedures.

Abbreviations

[0470] The following abbreviations are used in the Examples and other parts of the examples: [0471] CDCl.sub.3: Deuterated chloroform [0472] DBU: 1,8-Diazabicyclo[5,4,0]undec-7-ene [0473] DCM: Dichloromethane [0474] DIPEA: N,N-Diisopropylethylamine [0475] DMAP: 4-Dimethylaminopyridine [0476] DMF: N,N-Dimethylformamide [0477] EDCI: 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride [0478] Equiv: equivalent(s) [0479] EtOAc: Ethyl acetate [0480] h: Hour(s) [0481] HATU: (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate [0482] HPLC: High Performance Liquid Chromatography [0483] LCMS: Liquid chromatography-mass spectrometry [0484] M: Molar [0485] MeCN: Acetonitrile [0486] MeOH: Methanol [0487] Min(s): Minute(s) [0488] o/n: overnight [0489] r.t.: Room temperature [0490] Rt: Retention time [0491] sat.: Saturated [0492] TEA: Triethylamine [0493] TFA: Trifluoroacetic acid [0494] THF: Tetrahydrofuran [0495] vac: Vacuum

Analytical Methods:

[0496] Method A: Waters Acquity UPLC BEH C18 1.7 m, 2.130 mm; A=water+0.1% formic acid; B=MeCN; 45 C.; % B: 0.0 min 5% 0.60 mL/min, 0.05 min 5% 0.60 mL/min, 1.6 min 95% 0.60 mL/min, 2.25 min 95% 0.75 mL/min, 2.26 min 5% 0.60 mL/min, 2.60 min 5% 0.60 mL/min.

[0497] Method B: Waters Acquity UPLC BEH C18 1.7 m, 2.130 mm; A=water+10 mM ammonium bicarbonate; B=MeCN; 45 C.; % B: 0.0 min 5% 0.60 mL/min, 0.05 min 5% 0.60 mL/min, 1.6 min 95% 0.60 mL/min, 2.25 min 95% 0.75 mL/min, 2.26 min 5% 0.60 mL/min, 2.60 min 5% 0.60 mL/min.

[0498] Method C: Waters Acquity UPLC BEH C18 1.7 m, 2.130 mm; A=water+0.1% formic acid; B=MeCN; 45 C.; % B: 0.0 min 5% 0.35 mL/min, 0.05 min 5% 0.35 mL/min, 5.0 min 95% 0.35 mL/min, 6.5 min 95% 0.35 mL/min, 6.6 min 5% 0.35 mL/min, 9.0 min 5% 0.35 mL/min

[0499] Method D: Waters Acquity UPLC BEH C18 1.7 m, 2.130 mm; A=water+0.1% ammonia; B=MeCN; 45 C.; % B: 0.0 min 5% 0.35 mL/min, 0.05 min 5% 0.35 mL/min, 5.0 min 95% 0.35 mL/min, 6.5 min 95% 0.35 mL/min, 6.6 min 5% 0.35 mL/min, 9.0 min 5% 0.35 mL/min.

[0500] Method E: Waters Acquity UPLC BEH C18 1.7 m, 2.130 mm; A=water+10 mM ammonium bicarbonate; B=MeCN; 45 C.; % B: 0.0 min 5% 0.35 mL/min, 0.05 min 5% 0.35 mL/min, 5.0 min 95% 0.35 mL/min, 6.5 min 95% 0.35 mL/min, 6.6 min 5% 0.35 mL/min, 9.0 min 5% 0.35 mL/min.

[0501] Method F: Phenomenex Gemini NX C18 5 m, 1504.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 40 C.; % B: 0.0 min 5% 1.5 mL/min, 0.5 min 5% 1.5 mL/min, 7.5 min 95% 1.5 mL/min, 10.0 min 95% 1.5 mL/min, 10.1 min 5% 1.5 mL/min, 13.0 min 5% 1.5 mL/min.

[0502] Method G: Phenomenex Gemini NX C18 5 m, 1504.6 mm; A=water+0.1% formic acid; B=MeCN+0.1% formic acid; 40 C.; % B: 0.0 min 5% 1.5 mL/min, 0.5 min 5% 1.5 mL/min, 7.5 min 95% 1.5 mL/min, 10.0 min 95% 1.5 mL/min, 10.1 min 5% 1.5 mL/min, 13.0 min 5% 1.5 mL/min.

[0503] Method H: Waters Acquity UPLC BEH C18 1.7 m, 2.150 mm; A=water+0.1% formic acid; B=MeCN; 45 C.; % B: 0.0 min 5% 0.60 mL/min, 0.05 min 5% 0.60 mL/min, 1.6 min 95% 0.60 mL/min, 2.25 min 95% 0.75 mL/min, 2.26 min 5% 0.60 mL/min, 2.60 min 5% 0.60 mL/min.

[0504] Method I: Waters Acquity UPLC BEH C18 1.7 m, 2.150 mm; A=water+10 mM ammonium bicarbonate; B=MeCN; 45 C.; % B: 0.0 min 5% 0.60 mL/min, 0.05 min 5% 0.60 mL/min, 1.6 min 95% 0.60 mL/min, 2.25 min 95% 0.75 mL/min, 2.26 min 5% 0.60 mL/min, 2.60 min 5% 0.60 mL/min.

[0505] Method J: Waters Acquity CSH C18 1.7 m, 2.1100 mm; A=water+0.1% formic acid; B=MeCN+0.1% formic acid; 40 C.; % B: 0.0 min 5% 0.5 min 5%, 5.0 min 95%, 6.5 min, 6.6 min 5%, 9.0 min 5% 0.35 mL/min.

[0506] Method K: Waters Acquity CSH C18 1.7 m, 2.1100 mm; A=pH 9 ammonium bicarbonate 10 mM aqueous solution; B=MeCN; 40 C.; % B: 0.0 min 5% 0.5 min 5%, 5.0 min 95%, 6.5 min, 6.6 min 5%, 9.0 min 5% 0.35 mL/min.

[0507] Method L: Waters Sunfire C18 3.5 m, 4.6 mm50 mm; A=water+0.1% formic acid; B=MeCN; 45 C.; % B: 0.0 min 5% 2.25 mL/min 1.00 min 37.5% 2.20 mL/min 3.00 min 95% 2.2 mL/min 3.5 min 95% 2.3 mL/min 3.51 min 5% 2.3 mL/min 4.00 min 5% 2.25 mL/min.

Chemical Synthesis Example 1

General Procedure A: Chloroester Formation

##STR00169##

[0508] A mixture of carboxylic acid (1.0 equiv), sulfochoridate (1.4 equiv), NaHC.sub.3 (4.0 equiv), tetrabutyl ammonium hydrogen sulphate (10 mol %), DCM and water (1:1) were stirred vigorously at r.t. for 16 hours. It was then passed through a phase separator, washed with sat. NaHCO.sub.3 solution and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent concentrated in vacuo to afford the desired product without any further purification.

TABLE-US-00011 TABLE 10 Chemical Synthesis Example Name and structure Analytical data 1 tert-Butyl (chloromethyl) succinate [00170] Synthesised according to general procedure A, using 4- (tert-butoxy)-4-oxobutanoic acid (17.2 mmol) and chloromethyl chlorosulfate (24.1 mmol). .sup.1H-NMR (400 MHz, CDCl.sub.3) 4.97 (s, 2H), 1.72-2.09 (m, 4H), 0.69 (s, 9H). Colourless oil, 3.70 g, 98% 2 tert-Butyl (1-chloroethyl) succinate [00171] Synthesised according to general procedure A, using 4- (tert-butoxy)-4-oxobutanoic acid (2.9 mmol) and 1- chloroethyl sulfochloridate (4.0 mmol). .sup.1H-NMR (400 MHz, CDCl.sub.3) 6.55 (q, J = 6.0 Hz, 1H), 2.48- 2.70 (m, 4H), 1.78-1.86 (m, Colourless oil, 386 mg, 57% 3H), 1.37-1.45 (m, 9H). 3 Chloromethyl (R)-5-(1,2- dithiolan-3-yl)pentanoate [00172] Synthesised according to general procedure A, using lipoic acid (2.4 mmol) and chloromethyl chlorosulfate (3.4 mmol). The compound was used immediately in the next step without any further purification. Yellow oil, 240 mg, 37% 4 1-Chloroethyl 5-((R)-1,2- dithiolan-3-yl)pentanoate [00173] Synthesised according to general procedure A, using lipoic acid (3.8 mmol) and 1- chloroethyl sulfurochloridate (5.3 mmol). .sup.1H-NMR (400 MHz, CDCl.sub.3): 6.53 (q, J = 5.8 Hz, 1H), 3.48- 3.61 (m, 1H), 3.04-3.22 (m, 2H), 2.30-2.53 (m, 3H), 1.84- 1.96 (m, 1H), 1.77 (d, J = 6.0 Yellow oil, 617 mg, 34% Hz, 3H), 1.59-1.74 (m, 4H), 1.39-1.58 (m, 2H)

Chemical Synthesis Example 5

(R)-((5-(1,2-Dithiolan-3 yl)pentanoyl)oxy)methyl tert-butyl succinate

##STR00174##

[0509] To a solution of lipoic acid (4.17 g, 20.2 mmol) and DIPEA (8.78 mL, 50.4 mmol) in DMF (30 mL), tert-butyl (chloromethyl) succinate (3.74 g, 16.8 mmol) was added and the reaction mixture stirred at 50 C. for 16 h. The reaction was diluted with EtOAc and washed with 1:1 H.sub.2O-sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by flash chromatography (Biotage Isolera Four; 100 g Sfar cartridge) eluting with isohexane.fwdarw.60% EtOAc-isohexane to afford (R)-((5-(1,2-dithiolan-3-yl)pentanoyl)oxy)methyl tert-butyl succinate as a yellow oil (3.15 g, 48%). LCMS (Method A): Rt=2.05 min; [M+Na].sup.+=415.2. .sup.1H-NMR (400 MHz, CDCl.sub.3) 5.73 (s, 2H), 3.48-3.59 (m, 1H), 3.03-3.22 (m, 2H), 2.56-2.67 (m, 2H), 2.49-2.56 (m, 2H), 2.38-2.49 (m, 1H), 2.34 (t, J=7.3 Hz, 2H), 1.82-1.94 (m, 1H), 1.53-1.74 (m, 4H), 1.36-1.52 (m, 11H).

Chemical Synthesis Example 6

1-((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)ethyl tert-butyl succinate

##STR00175##

[0510] To a solution of lipoic acid (270 mg, 1.31 mmol) and DIPEA (0.68 mL, 3.90 mmol) in DMF (2.0 mL), tert-butyl (1-chloroethyl) succinate (386 mg, 1.60 mmol) was added and the reaction mixture stirred at 50 C. for 16 h. The reaction mixture was diluted with EtOAc and washed with 1:1 H.sub.2O-sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by flash chromatography (Biotage Isolera Four; 100 g Sfar cartridge) eluting with 75% EtOAc-isohexane to afford 1-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)ethyl tert-butyl succinate as a yellow oil (114 mg, 21%). LCMS (Method A): Rt=2.17 min; [M+H].sup.+=407.2. .sup.1H NMR (400 MHz, CDCl.sub.3) 6.84-6.89 (m, 1H), 3.55 (dt, J=14.7, 6.4 Hz, 1H), 3.09-3.20 (m, 1H), 2.44-2.63 (m, 6H), 2.29-2.33 (m, 1H), 1.86-1.94 (m, 1H), 1.60-1.71 (m, 2H), 1.51-1.59 (m, 3H), 1.46 (dd, J=5.5, 1.4 Hz, 3H), 1.43-1.44 (m, 10H).

Chemical Synthesis Example 7

(R)-4-(((5-(1,2-Dithiolan-3-yl)pentanoyl)oxy)methoxy)-4-oxobutanoic acid

##STR00176##

[0511] To a solution of (R)-((5-(1,2-Dithiolan-3-yl)pentanoyl)oxy)methyl tert-butyl succinate (110 mg, 0.28 mmol) in DCM (3.0 mL), was added TFA (400 L, 5.23 mmol) and the reaction mixture stirred at r.t. for 3 h. The pH of the reaction mixture was then adjusted to pH 4 with sat. NaHCO.sub.3(aq) sol. and the reaction mixture passed through a phase separator. The filtrate was evaporated in vacuo to afford (R)-4-(((5-(1,2-dithiolan-3-yl)pentanoyl)oxy)methoxy)-4-oxobutanoic acid as a colorless oil (80.0 mg, 85%). This was used without further purification. LCMS (Method A): Rt=1.66 min; [M+NH.sub.4].sup.+=354.2.

Chemical Synthesis Example 8

4-(1-((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)ethoxy)-4-oxobutanoic acid

##STR00177##

[0512] To a solution of 1-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)ethyl tert-butyl succinate (114 mg, 0.280 mmol) in DCM (4.0 mL), was added TFA (400 L, 5.96 mmol) and the reaction mixture stirred at r.t. for 3 h. The pH of the reaction mixture was then adjusted to pH 4 with sat. NaHCO.sub.3(aq) and the reaction mixture passed through a phase separator. The filtrate was evaporated in vacuo to afford 4-(1-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)ethoxy)-4-oxobutanoic acid as a colorless oil (27.0 mg, 27%). This was used without further purification.

[0513] LCMS (Method A): Rt=1.78 min; [M+NH.sub.4].sup.+=368.1; [M+Na].sup.+=373.1.

Chemical Synthesis Example 9

Benzyl (2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carboxylate

##STR00178##

[0514] A mixture of benzyl (2S,5R)-5-amino-2-methylpiperidine-1-carboxylate (2.00 g, 8.05 mmol), potassium carbonate (3.56 g, 25.8 mmol) and 4-chloro-7H-pyrrolo(2,3,d)pyrimidine (1.30 g, 8.48 mmol) in water (20 mL) was stirred at 90 C. for 4 days. The reaction mixture was diluted with EtOAc (20 mL) and the organic phase separated, washed with water (10 mL) and sat. brine solution (10 mL), dried (MgSO.sub.4), filtered and evaporated in vacuo. The crude product was purified by flash chromatography (Biotage SP1; 50 g Sfar cartridge) eluting with isohexane.fwdarw.acetone to afford benzyl (2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carboxylate (2.87 g, 98%) as a light brown solid. LCMS (Method B): Rt=1.68 min; [M+H].sup.+=366.2. .sup.1H-NMR (400 MHz, DMSO-D6) 11.50 (s, 1H), 8.09 (s, 1H), 7.26-7.46 (m, 5H), 7.20 (d, J=8.2 Hz, 1H), 7.08 (t, J=2.7 Hz, 1H), 6.47-6.61 (m, 1H), 4.91-5.24 (m, 2H), 3.94-4.49 (m, 3H), 2.71 (br s, 1H), 1.44-1.93 (m, 4H), 1.07-1.26 (m, 3H).

Chemical Synthesis Example 10

N-((3R,6S)-6-Methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

##STR00179##

[0515] Benzyl (2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidine-1-carboxylate (7.08 g, 19.4 mmol) was dissolved in ethanol (40 mL) and palladium 10% wt on carbon (0.5 g) added. The solution was placed under N.sub.2 in a large parr flask (500 mL). The resulting mixture was shaken on a parr apparatus (40 psi of H.sub.2 at r.t. for 5 h). The reaction mixture was filtered through a pad of Celite, and the cake was washed with EtOAc. The filtrate was concentrated in vacuo to afford N-((3R,6S)-6-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (4.34 g, 97%) as an off-white solid. LCMS (Method B): Rt=1.14 min; [M+H].sup.+=232.2. .sup.1H-NMR (400 MHz, DMSO-D6) 11.44 (s, 1H), 8.03 (s, 1H), 7.03 (d, J=3.6 Hz, 1H), 6.82 (d, J=7.3 Hz, 1H), 6.60 (d, J=2.3 Hz, 1H), 4.09-4.11 (m, 1H), 2.87-2.97 (m, 1H), 2.71-2.84 (m, 1H), 2.49-2.63 (m, 1H), 2.07 (br s, 1H), 1.80-1.93 (m, 1H), 1.58 (tt, J=12.7, 4.0 Hz, 1H), 1.34-1.45 (m, 1H), 1.27 (m, 1H), 0.96 (d, J=6.4 Hz, 3H).

Chemical Synthesis Example 11

1-((2S,5R)-5-((7H-Pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one

##STR00180##

[0516] To a stirred solution of N-((3R,6S)-6-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (4.34 g, 18.8 mmol) in THF (80 mL) and sat. NaHCO.sub.3 solution (80 mL) was added acryloyl chloride (1.82 mL, 22.5 mmol) dropwise at 0 C. After addition, the resulting mixture was stirred at 0 C. for 2 h. The reaction mixture was diluted with water and extracted with EtOAc (350 mL). The combined organics were washed with sat. brine solution (30 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by flash chromatography (Biotage SP1; 100 g Sfar cartridge) eluting with DCM.fwdarw.10% MeOH-DCM to afford 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (2.24 g, 42%) as a white solid. LCMS (Method C): Rt=2.27 min; [M+H].sup.+=286.2. LCMS (Method E): Rt=2.38 min; [M+H].sup.+=286.2. .sup.1H-NMR (400 MHz, DMSO-D6) 11.53 (s, 1H), 8.07 (d, J=11.9 Hz, 1H), 7.24 (dd, J=18.3, 7.3 Hz, 1H), 7.04-7.05 (m, 1H), 6.69-6.80 (m, 1H), 6.46-6.57 (m, 1H), 6.05 (dd, J=19.2, 2.3 Hz, 1H), 5.63 (dd, J=12.8, 2.3 Hz, 1H), 4.76 (br s, 0.5H), 4.51 (br s, 0.5H), 4.32 (br s, 0.5H), 3.89-4.22 (m, 1.5H), 2.81-3.01 (m, 0.5H), 2.48-2.67 (m, 0.5H), 1.51-1.90 (m, 4H), 1.05-1.26 (m, 3H).

Chemical Synthesis Examples 12, 13 & 14

(4-(((3R,6S)-1-Acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate (12), (4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoate (13), and (4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate (14)

##STR00181##

[0517] To a solution of 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (200 mg, 0.70 mmol) in DMF (2 mL) at 0 C., sodium hydride (60% dispersion in oil, 33.6 mg, 0.84 mmol) was added and the mixture stirred at 0 C. for 15 min. Chloromethyl 5-[(3R)-dithiolan-3-yl]pentanoate (267 mg, 1.05 mmol) was added and the reaction stirred at 0 C. for 1 h. The reaction was diluted with EtOAc (10 mL) and washed with 1:1 water-sat. brine solution (515 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo. The crude product was purified by flash chromatography (Biotage SP1; 25 g Sfar cartridge) eluting with isohexane.fwdarw.acetone followed by preparative reversed-phase HPLC to afford (4-(((3R,6S)-1-Acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate, (4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoate, and (4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate.

[0518] (4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate (21.0 mg, 6%) as a light yellow oil. LCMS (Method C): Rt=4.27 min; [M+H].sup.+=504.1. LCMS (Method E): Rt=4.93 min; [M+H].sup.+=504.1. .sup.1H-NMR (400 MHz, DMSO-D6) 8.16 (d, J=12.4 Hz, 1H), 7.47 (dd, J=15.8, 7.1 Hz, 1H), 7.22 (d, J=3.7 Hz, 1H), 6.65-6.88 (m, 1H), 6.61 (d, J=3.7 Hz, 1H), 5.97-6.14 (m, 3H), 5.63 (dd, J=10.3, 2.1 Hz, 1H), 4.76 (br s, 0.5H), 4.51 (d, J=9.2 Hz, 0.5H), 4.33 (br s, 0.5H), 3.81-4.22 (m, 1.5H), 3.39-3.59 (m, 1H), 2.74-3.19 (m, 2.5H), 2.49-2.68 (m, 0.5H), 2.15-2.37 (m, 3H), 1.35-1.91 (m, 9H), 1.00-1.35 (m, 5H).

[0519] (4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoate and (4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate (2.8 mg, 1%) as a white solid. LCMS (Method C): Rt=3.35 min; [M+H].sup.+=520.1. LCMS (Method E): Rt=3.99 min; [M+H].sup.+=520.2.

Chemical Synthesis Example 15

1-(4-(((3R,6S)-1-Acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((R)-1,2-dithiolan-3-yl)pentan-1-one

##STR00182##

[0520] Lipoic acid (75.9 mg, 0.37 mmol) and 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (94.0 mg, 0.49 mmol) were dissolved in DMF (1.0 mL) and stirred at r.t. for 1 h. 4-(Dimethylamino)pyridine (29.9 mg, 0.25 mmol) and 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (70.0 mg, 0.25 mmol) were added and the reaction mixture stirred at r.t. for 16 h. The reaction mixture was diluted with EtOAc (10 mL) and washed with 1:1 water-sat. brine solution (515 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by flash chromatography (Biotage SP1; 10 g Sfar cartridge) eluting with isohexane.fwdarw.80% acetone-isohexane to afford 1-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((R)-1,2-dithiolan-3-yl)pentan-1-one (52 mg, 45%) as a light yellow solid. LCMS (Method C): Rt=5.19 min; [M+H].sup.+=474.2. LCMS (Method E): Rt=5.45 min; [M+H].sup.+=474.2. .sup.1H-NMR (400 MHz, DMSO-D6) 8.29 (d, J=8.2 Hz, 1H), 7.57-7.66 (m, 2H), 6.61-6.90 (m, 2H), 6.05 (dd, J=16.7, 2.5 Hz, 1H), 6.63 (dd, J=10.3, 2.5 Hz, 1H), 4.76 (br s, 0.5H), 4.41-4.61 (m, 0.5H), 4.34 (br s, 0.5H), 3.86-4.20 (m, 1.5H), 3.51-3.74 (m, 1H), 3.34-3.51 (m, 2H), 2.99-3.21 (m, 2H), 2.92 (t, J=11.4 Hz, 0.5H), 2.50-2.70 (m, 0.5H), 2.31-2.43 (m, 1H), 1.36-1.94 (m, 11H), 1.06-1.30 (m, 3H).

Chemical Synthesis Examples 16 & 17

1-(4-(((3R,6S)-1-Acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentan-1-one (16) and 1-(4-(((3R,6S)-1-Acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentan-1-one (17)

##STR00183##

[0521] 5-[(3R)-2-Oxodithiolan-3-yl]pentanoic acid (105 mg, 0.47 mmol) and 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (121 mg, 0.63 mmol) were dissolved in DMF (1.0 mL) and stirred at r.t. for 1 h. 4-(Dimethylamino)pyridine (38.5 mg, 0.32 mmol) and 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (90.0 mg, 0.32 mmol) were added and the mixture stirred at r.t for 16 hours. The reaction mixture was diluted with EtOAc (10 mL) and washed with 1:1 water-sat. brine solution (515 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by flash chromatography (Biotage SP1; 10 g Sfar cartridge) eluting with isohexane.fwdarw.acetone to afford 1-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentan-1-one and 1-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentan-1-one (30 mg, 19%) as a white solid. LCMS (Method C): Rt=4.01 min; [M+H].sup.+=490.2. LCMS (Method E): Rt=4.37 mins; [M+H].sup.+=490.2.

Chemical Synthesis Example 18

Methyl 4-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate

##STR00184##

[0522] Mono-methyl hydrogen succinate (69.4 mg, 0.53 mmol) and 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (134 mg, 0.70 mmol) were dissolved in DMF (1.0 mL) and stirred at r.t. for 1 h. 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (100 mg, 0.35 mmol) and 4-(dimethylamino)pyridine (43.0 mg, 0.35 mmol) were added and the mixture was stirred at r.t for 16 hours. The reaction mixture was diluted with EtOAc (5 mL) and washed with 1:1 water-sat. brine solution (510 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by flash chromatography (Biotage SP1; 10 g Sfar cartridge) eluting with isohexane.fwdarw.acetone to afford methyl 4-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate (87.0 mg, 62%) as a white solid. LCMS (Method C): Rt=3.85 min; [M+H].sup.+=400.3. LCMS (Method E): Rt=4.21 min; [M+H].sup.+=400.2. .sup.1H-NMR (400 MHz, DMSO-D6) 8.30 (d, J=8.2 Hz, 1H), 7.54-7.84 (m, 2H), 6.58-6.99 (m, 2H), 6.05 (dd, J=16.4, 2.3 Hz, 1H), 5.63 (dd, J=10.4, 2.3 Hz, 1H), 4.76 (br s, 0.5H), 4.44-4.66 (m, 0.5H), 4.34 (br s, 0.5H), 3.90-4.24 (m, 1.5H), 3.65 (t, J=12.8 Hz, 2H), 3.57 (s, 3H), 2.80-3.06 (m, 0.5H), 2.72 (t, J=6.4 Hz, 2H), 2.50-2.65 (m, 0.5H), 1.47-1.94 (m, 4H), 1.01-1.34 (m, 3H).

Chemical Synthesis Example 19

((4-(4-(((3R,6S)-1-Acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoyl)oxy)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate

##STR00185##

[0523] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (68.3 mg, 0.36 mmol) was suspended in anhydrous DCM (2.0 mL) and DMF (0.5 mL), then 4-[5-[(3R)-dithiolan-3-yl]pentanoyloxymethoxy]-4-oxo-butanoic acid (48.0 mg, 0.14 mmol) was added and the mixture stirred at r.t. for 1 h. 4-(Dimethylamino)pyridine (17.4 mg, 0.14 mmol) and 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (40.6 mg, 0.14 mmol) were added and the mixture stirred at r.t. for 16 hours. The reaction mixture was evaporated in vacuo and the residue re-dissolved in DMSO and purified by preparative reversed-phase HPLC to afford ((4-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoyl)oxy)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate (6.3 mg, 7%) as a white solid. LCMS (Method C): Rt=5.21 min; [M+H].sup.+=604.2. LCMS (Method E): Rt=5.35 min; [M+H].sup.+=604.2.

Chemical Synthesis Example 20

Chloromethyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate

##STR00186##

[0524] To a solution of 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (300 mg, 1.05 mmol) in THF (3.0 mL) at 0 C., DIPEA (0.22 mL, 1.26 mmol) was added, and the reaction mixture stirred for 15 min at 0 C. Chloromethyl chloroformate (0.12 mL, 1.26 mmol) was added and the reaction mixture stirred at 0 C. for 1 h and then at r.t. for 1 h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage SP1; 25 g Sfar cartridge) eluting with isohexane.fwdarw.acetone to afford chloromethyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (180 mg, 39%) as a white solid. LCMS (Method B): Rt=1.48 min; [M+H].sup.+=378.2. .sup.1H-NMR (400 MHz, DMSO-D6) 8.30 (br s, 1H), 7.70 (br s, 1H), 7.52 (d, J=3.7 Hz, 1H), 6.87 (d, J=4.1 Hz, 1H), 6.64-6.84 (m, 1H), 6.14 (s, 2H), 6.05 (dd, J=16.9, 2.3 Hz, 1H), 5.64 (dd, J=10.5, 2.3 Hz, 1H), 4.76 (br s, 0.5H), 4.53 (br s, 0.5H), 4.33 (br s, 0.5H), 3.83-4.21 (m, 1.5H), 2.93 (t, J=11.7 Hz, 0.5H), 2.51-2.75 (m, 0.5H), 1.55-1.87 (m, 4H), 1.15 (dd, J=21.8, 6.6 Hz, 3H).

Chemical Synthesis Example 21

((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)methyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate

##STR00187##

[0525] To a solution of chloromethyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (90.0 mg, 0.16 mmol) in DMF (1.0 mL), (R)-5-(1,2-dithiolan-3-yl)pentanoic acid (66 mg, 0.31 mmol) and DIPEA (97 L, 0.55 mmol) were added and the reaction mixture was stirred at r.t. for 16 hours. The reaction mixture was diluted with EtOAc (5 mL) and washed with 1:1 water-sat. brine solution (510 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The residue was re-dissolved in DMSO and purified by preparative reversed-phase HPLC to afford ((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)methyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (7.2 mg, 8%) as a white solid. LCMS (Method C): Rt=4.73 min; [M+H].sup.+=548.1. LCMS (Method E): Rt=4.91 min; [M+H].sup.+=548.1. .sup.1H-NMR (400 MHz, DMSO-D6) 8.18-8.38 (m, 1H), 7.58-7.74 (m, 1H), 7.46 (d, J=3.7 Hz, 1H), 6.57-6.89 (m, 2H), 6.05 (dd, J=16.7, 2.5 Hz, 1H), 5.95 (s, 2H), 5.64 (dd, J=10.5, 2.3 Hz, 1H), 4.76 (br s, 0.5H), 4.43-4.62 (m, 0.5H), 4.26-4.41 (m, 0.5H), 3.90-4.20 (m, 1.5H), 3.40-3.64 (m, 0.5H), 2.83-3.16 (m, 2H), 2.50-2.67 (m, 0.5H), 2.39 (t, J=7.3 Hz, 2H), 2.29 (m, 1H), 1.00-1.93 (m, 15H).

Chemical Synthesis Examples 22 & 23

1-(4-(((3R,6S)-1-Acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isopropyl carbonate (22) and isopropyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (23)

##STR00188##

[0526] To a solution of 1-((2S,5R)-5-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (60.0 mg, 0.21 mmol) in DMF (2.0 mL) at 0 C., sodium hydride (10.0 mg, 0.25 mmol, 60% dispersion in oil) was added and the mixture was left stirring at r.t. for 15 min. 1-chloroethyl isopropyl carbonate (48 L, 0.32 mmol) was added and the reaction mixture stirred at r.t. for 30 min. The reaction mixture was diluted with EtOAc (5 mL) and washed with 1:1 water-sat. brine solution (510 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo. The crude product was purified by flash chromatography (Biotage SP1; 10 g Sfar cartridge) eluting with isohexane.fwdarw.acetone to afford 1-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isopropyl carbonate and isopropyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate.

[0527] 1-(4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isopropyl carbonate (15.0 mg, 17%), as an off-white solid. LCMS (Method C): Rt=3.61 min; [M+H].sup.+=416.3. LCMS (Method E): Rt=4.46 min; [M+H].sup.+=416.2.

[0528] Isopropyl 4-(((3R,6S)-1-acryloyl-6-methylpiperidin-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (26.0 mg, 33%) was obtained as white gum. LCMS (Method C): Rt=3.60 min; [M+H].sup.+=372.3. LCMS (Method E): Rt=4.14 min; [M+H].sup.+=372.2. .sup.1H-NMR (400 MHz, DMSO-D6) 8.26 (d, J=11.0 Hz, 1H), 7.58-7.71 (m, 1H), 7.47 (d, J=3.9 Hz, 1H), 6.67-6.87 (m, 2H), 6.05 (dd, J=16.9, 2.3 Hz, 1H), 5.64 (dd, J=10.5, 1.8 Hz, 1H), 5.10 (sep, J=6.4 Hz, 1H), 4.75 (br s, 0.5H), 4.56 (br s, 0.5H), 4.33 (br s, 0.5H), 3.89-4.16 (m, 1.5H), 2.81-3.02 (m, 0.5H), 2.49-2.70 (m, 0.5H), 1.55-1.88 (m, 4H), 1.29-1.40 (m, 6H), 1.11-1.23 (m, 3H).

Chemical Synthesis Example 24

(4-(((3R,4R)-1-(2-Cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate

##STR00189##

[0529] To a solution of Tofacitinib (50 mg, 0.16 mmol) in DMF (2.0 mL), sodium hydride (60% dispersion in oil, 7.0 mg, 0.18 mmol) was added and the reaction mixture stirred at r.t. for 15 min. Chloromethyl-(R)-5-(1,2-dithiolan-3-yl)pentanoate (75 mg, 2.9 mmol) was then added and the reaction mixture stirred at 0 C. for 15 min. The reaction mixture was partitioned between water (5 mL) and EtOAc (5 mL) and the layers were separated. The organic phase was washed successively with sat. NaHCO.sub.3(aq) and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford (4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate as a white solid (3.8 mg, 5%). LCMS (Method C): Rt=5.12 min; [M+H].sup.+=531.2, LCMS (Method E): Rt=5.39 min; [M+H].sup.+=531.2.

Chemical Synthesis Example 25

3-((3R,4R)-3-((7-(5-((R)-1,2-Dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(methyl)amino)-4-methylpiperidin-1-yl)-3-oxopropanenitrile

##STR00190##

[0530] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (61 mg, 0.32 mmol) was dissolved in anhydrous DCM (2.0 mL), lipoic acid (26 mg, 0.13 mmol) was added and the mixture stirred at r.t. for 1 h. Separately, DMAP (16 mg, 0.13 mmol) was dissolved in anhydrous DCM (2.0 mL), Tofacitinib (40 mg, 0.13 mmol) was added and the mixture stirred at r.t. for 10 min. The two solutions were combined and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 70% methyl acetate in pentane. The solvent was evaporated by flow of N.sub.2 to afford 3-((3R,4R)-3-((7-(5-((R)-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(methyl)amino)-4-methylpiperidin-1-yl)-3-oxopropanenitrile as a yellow solid (20.3 mg, 32%). LCMS (Method C): Rt=5.30 min; [M+H].sup.+=501.2, LCMS (Method E): Rt=5.55 min; [M+H].sup.+=501.2.

Chemical Synthesis Example 26

((4-(4-(((3R,4R)-1-(2-Cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoyl)oxy)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate

##STR00191##

[0531] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (132 mg, 0.69 mmol) was dissolved in anhydrous DCM (2.0 mL), (R)-4-(((5-(1,2-dithiolan-3-yl)pentanoyl)oxy)methoxy)-4-oxobutanoic acid (93 mg, 0.28 mmol) was then added and the mixture was stirred at r.t. for 1 h. Separately, DMAP (33 mg, 0.28 mmol) was dissolved in anhydrous DCM (2.0 mL), Tofacitinib (86 mg, 0.28 mmol) was added, the mixture was stirred at r.t. for 10 min. The two solutions were combined and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by preparative reversed-phase HPLC to afford ((4-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoyl)oxy)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate as a white solid (2.0 mg, 2%). LCMS (Method C): Rt=5.30 min; [M+H].sup.+=631.2, LCMS (Method E): Rt=5.40 min; [M+H].sup.+=631.2.

Chemical Synthesis Example 27

1-((4-(4-(((3R,4R)-1-(2-Cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo [2,3-d]pyrimidin-7-yl)-4-oxobutanoyl)oxy)ethyl 5-((R)-1,2-dithiolan-3-yl)pentanoate

##STR00192##

[0532] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (37 mg 0.19 mmol) was dissolved in anhydrous DCM (2.0 mL), 4-(1-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)ethoxy)-4-oxobutanoic acid (27 mg, 77 mol) was added and the reaction mixture stirred at r.t. for 1 h. Separately, DMAP (9.4 mg, 77 mol) was dissolved in anhydrous DCM (2.0 mL), Tofacitinib (24 mg, 77 mol) was added, the mixture was stirred at r.t. for 10 min. The two solutions were combined and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by preparative reversed-phase HPLC to afford 1-((4-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo [2,3-d]pyrimidin-7-yl)-4-oxobutanoyl)oxy)ethyl 5-((R)-1,2-dithiolan-3-yl)pentanoate as a white solid (1.0 mg, 2%). LCMS (Method C): Rt=5.52 min; [M+H].sup.+=645.3, LCMS (Method E): Rt=5.64 min; [M+H].sup.+=645.3.

Chemical Synthesis Example 28

Methyl 4-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate

##STR00193##

[0533] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (46 mg, 0.24 mmol) was dissolved in anhydrous DCM (2.0 mL), mono-methyl hydrogen succinate (13 mg, 96 mol) was added and the mixture stirred at r.t. for 1 h. Separately, DMAP (12 mg, 96 mol) was dissolved in anhydrous DCM (2.0 mL), Tofacitinib (30 mg, 96 mol) was added, the mixture was stirred at r.t. for 10 min. The two solutions were combined and the reaction mixture stirred at r.t. for 16h. The solvent was evaporated in vacuo and the crude product was purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 100% EtOAc, to afford methyl 4-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate as a beige oil (19.8 mg, 48%). LCMS (Method C): Rt=3.98 min; [M+H].sup.+=427.3, LCMS (Method D): Rt=3.29 min; [M+H].sup.+=427.3.

Chemical Synthesis Example 29

Ethyl 4-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate

##STR00194##

[0534] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (46 mg, 0.24 mmol) was dissolved in anhydrous DCM (2.0 mL), mono-ethyl succinate (14 mg, 96 mol) was added and the reaction mixture stirred at r.t. for 1 h. Separately, DMAP (12 mg, 96 mol) was dissolved in anhydrous DCM (2.0 mL), Tofacitinib (30 mg, 96 mol) was added and the reaction mixture was stirred at r.t. for 10 min. The two solutions were combined, and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 100% EtOAc, to afford ethyl 4-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidin-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate as a colorless oil (11.5 mg, 27%). LCMS (Method C): Rt=4.33 min; [M+H].sup.+=441.2, LCMS (Method D): Rt=4.64 min; [M+H].sup.+=441.2.

Chemical Synthesis Example 30

1-(4-(Methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl 5-((R)-1,2-dithiolan-3-yl)pentanoate

##STR00195##

[0535] To a solution of Oclacitinib (40 mg, 0.12 mmol) in DMF (1.5 mL), sodium hydride (60% dispersion in oil, 4.7 mg, 0.12 mmol) was added and the reaction mixture stirred at r.t. for 15 min. 1-Chloroethyl 5-[(R)-1,2-dithiolan-3-yl]pentanoate (38 mg, 0.14 mmol) and NaI (18 mg, 0.12 mmol) were added and the reaction mixture stirred at 50 C. for 16 hours. The reaction mixture was partitioned between water (10 mL) and EtOAc (10 mL). The layers were separated and the organic phase washed successively with sat. NaHCO.sub.3(aq) and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford 1-(4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl 5-((R)-1,2-dithiolan-3-yl)pentanoate as a white solid (1.9 mg, 3%). LCMS (Method C): Rt=4.16 min; [M+H].sup.+=570.1, LCMS (Method E): Rt=5.26 min; [M+H].sup.+=570.1

Chemical Synthesis Examples 31, 32, & 33

(4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate (31), (4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate (32), and (4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoate (33)

##STR00196##

[0536] To a solution of Oclacitinib (20 mg, 59 mol) in DMF (1.5 mL), sodium hydride (60% dispersion in oil, 2.4 mg, 59 mol) was added and the reaction mixture stirred at r.t. for 15 min. Chloromethyl-(R)-5-(1,2-dithiolan-3-yl)pentanoate (33 mg, 0.13 mmol) was added and the reaction mixture stirred at r.t for 2 h. The reaction mixture was partitioned between water (5 mL) and EtOAc (5 mL). The layers were separated and the organic phase washed successively with sat. NaHCO.sub.3(aq) and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford (4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate, (4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate, and (4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoate.

[0537] (4-(Methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyri-midin-7-yl)methyl 5-((R)-1,2-dithiolan-3-yl)pentanoate as a white solid (5.6 mg, 27%). LCMS (Method C): Rt=4.08 min; [M+H].sup.+=556.1. LCMS (Method D): Rt=5.04 min; [M+H].sup.+=556.1. .sup.1H-NMR (400 MHz, DMSO-D6) 8.08-8.24 (m, 1H), 7.29 (d, J=3.7 Hz, 1H), 6.87 (q, J =5.0 Hz, 1H), 6.64 (d, J=3.2 Hz, 1H), 6.09-6.15 (m, 2H), 4.64 (d, J=9.6 Hz, 1H), 3.46-3.57 (m, 2H), 3.04-3.20 (m, 4H), 2.94-3.00 (m, 2H), 2.54-2.59 (m, 3H), 2.28-2.35 (m, 2H), 2.03-2.05 (m, 2H), 1.24-1.88 (m, 14H).

[0538] (4-(Methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate and (4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoate as a white solid (1.1 mg, 3%). LCMS (Method C): Rt=3.23 min; [M+H].sup.+=572.1. LCMS (Method D): Rt=4.12 min; [M+H].sup.+=572.1.

Chemical Synthesis Example 34

1-((1r,4r)-4-((7-(5-((R)-1,2-Dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(methyl)amino)cyclohexyl)-N-methylmethanesulfonamide

##STR00197##

[0539] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (57 mg, 0.30 mmol) was dissolved in anhydrous DCM (2.0 mL), lipoic acid (25 mg, 0.12 mmol) was added and the reaction mixture stirred at r.t. for 1 h. Separately, DMAP (15 mg, 0.12 mmol) was dissolved in anhydrous DCM (2.0 mL), Oclacitinib (40 mg, 0.12 mmol) was added, and the reaction mixture stirred at r.t. for 10 min. The two solutions were combined, and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 70% methyl acetate in pentane, followed by evaporation by N.sub.2 to afford 1-((1R,4r)-4-((7-(5-((R)-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(methyl)amino)cyclohexyl)-N-methylmethanesulfonamide as a yellow solid (26.7 mg, 43%). LCMS (Method C): Rt=5.11 min; [M+H].sup.+=526.2. LCMS (Method E): Rt=5.64 min; [M+H].sup.+=526.2.

Chemical Synthesis Examples 35 & 36

N-methyl-1-((1R,4r)-4-(methyl(7-(5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)methanesulfonamide (35) and N-methyl-1-((1R,4r)-4-(methyl(7-(5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)methanesulfonamide (36)

##STR00198##

[0540] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (71 mg, 0.37 mmol) was dissolved in anhydrous DCM (2.0 mL), 5-[(3R)-2-oxido-1,2-dithiolan-3-yl]pentanoic acid (33 mg, 0.15 mmol) was added and the reaction mixture stirred at r.t. for 1 h. Separately, DMAP (18 mg, 0.15 mmol) was dissolved in anhydrous DCM (2.0 mL), Oclacitinib (50 mg, 0.15 mmol) was added and the reaction mixture stirred at r.t. for 10 min. The two solutions were combined and the reaction mixture stirred at r.t. for 16h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 100% EtOAc to afford N-methyl-1-((1R,4r)-4-(methyl(7-(5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)methanesulfonamide and N-methyl-1-((1R,4r)-4-(methyl(7-(5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)methanesulfonamide as a colorless oil (13.3 mg, 17%). LCMS (Method C): Rt=3.85 min; [M+H].sup.+=542.2. LCMS (Method E): Rt=4.55 min; [M+H].sup.+=542.2.

Chemical Synthesis Example 37

Ethyl 4-(4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo [2,3-d]pyrimidin-7-yl)-4-oxobutanoate

##STR00199##

[0541] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (71 mg, 0.37 mmol) was dissolved in anhydrous DCM (2.0 mL), mono-ethyl succinate (22 mg, 0.15 mmol) was then added and the reaction mixture stirred at r.t. for 1 h. Separately, DMAP (18 mg, 0.15 mmol) was dissolved in anhydrous DCM (2.0 mL), Oclacitinib (50 mg, 0.15 mmol) was added and the reaction mixture stirred at r.t. for 10 min. The two solutions were combined and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 80% EtOAc in isohexane, to afford ethyl 4-(4-(methyl((1r,4r)-4-((N-methylsulfamoyl)methyl)cyclohexyl)amino)-7H-pyrrolo [2,3-d]pyrimidin-7-yl)-4-oxobutanoate as a white solid (14.3 mg, 21%). LCMS (Method C): Rt=4.12 min; [M+H].sup.+=466.2. LCMS (Method E): Rt=4.72 min; [M+H].sup.+=466.2. .sup.1H-NMR (400 MHz, DMSO-D6) 8.30-8.34 (m, 1H), 7.67 (d, J=4.1 Hz, 1H), 6.87 (q, J=4.9 Hz, 2H), 4.02-4.09 (m, 2H), 3.70 (t, J=6.4 Hz, 2H), 3.17 (d, J=7.8 Hz, 3H), 2.95 (d, J=6.0 Hz, 2H), 2.74 (t, J=6.4 Hz, 2H), 2.58-2.61 (m, 3H), 1.99-2.06 (m, 2H), 1.82-1.91 (m, 1H), 1.68-1.75 (m, 4H), 1.23-1.34 (m, 2H), 1.12-1.21 (m, 4H).

Chemical Synthesis Examples 38 & 39

1-(4-(1-((R)-2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl 5-((R)-1,2-dithiolan-3-yl)pentanoate (38) and (S)-3-(4-(7-(5-((R)-1,2-Dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile (39)

##STR00200##

[0542] To a solution of Ruxolitinib (50 mg, 0.16 mmol) in DMF (2.0 mL), sodium hydride (60% dispersion in oil, 7.5 mg, 0.19 mmol) was added and the reaction mixture stirred at r.t. for 15 min. 1-Chloroethyl 5-[-1,2-dithiolan-3-yl]pentanoate (110 mg, 0.41 mmol) and NaI (25 mg, 0.16 mmol) were then added and the reaction mixture stirred at 50 C. for 16 h. The reaction mixture was partitioned between water (10 mL) and EtOAc (10 mL). The layers were separated and the organic phase washed successively with sat. NaHCO.sub.3(aq) and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford 1-(4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl 5-((R)-1,2-dithiolan-3-yl)pentanoate and (S)-3-(4-(7-(5-((R)-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile.

[0543] 1-(4-(1-((R)-2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl-5-((R)-1,2-dithiolan-3-yl)pentanoate as an off-white solid (2.2 mg, 3%). LCMS (Method C): Rt=5.59 min; [M+H].sup.+=539.2. LCMS (Method D): Rt=5.71 min; [M+H].sup.+=539.2.

[0544] (S)-3-(4-(7-(5-((R)-1,2-Dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyra-zol-1-yl)-3-cyclopentylpropanenitrile as a white solid (7.0 mg, 9%). LCMS (Method C): Rt =6.04 min; [M+H].sup.+=495.2. LCMS (Method E): Rt=6.03 min; [M+H].sup.+=495.2. .sup.1H-NMR (400 MHz, DMSO-D6) 8.89-8.94 (m, 2H), 8.40 (d, J=22.0 Hz, 1H), 8.12 (d, J=4.1 Hz, 1H), 7.32 (d, J=4.1 Hz, 1H), 4.55 (td, J=9.6, 4.5 Hz, 1H), 3.64-3.73 (m, 1H), 3.48-3.56 (m, 2H), 3.10-3.29 (m, 4H), 2.33-2.47 (m, 2H), 1.88-1.95 (m, 1H), 1.71-1.87 (m, 4H), 1.47-1.69 (m, 6H), 1.12-1.45 (m, 4H).

Chemical Synthesis Examples 40, 41, & 42

[4-[(1S)-1-(2-Cyano-1-cyclopentyl-ethyl)pyrazol-4-yl]pyrrolo[2,3-d]pyrimidin-7-yl]methyl 5-[(3R)-dithiolan-3-yl]pentanoate (40), (4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl) pentanoate (41), and (4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate (42)

##STR00201##

[0545] To a solution of Ruxolitinib (60 mg, 0.20 mmol) in DMF (2.0 mL), sodium hydride (60% dispersion in oil, 12 mg, 0.30 mmol) was added and the reaction mixture stirred at r.t. for 15 min. Chloromethyl-(R)-5-(1,2-dithiolan-3-yl)pentanoate (100 mg, 0.39 mmol) was then added and the reaction mixture stirred at r.t for 2 h. The reaction mixture was partitioned between water and EtOAc. The layers were separated and the organic phase washed successively with sat. NaHCO.sub.3(aq) and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford [4-[(1S)-1-(2-cyano-1-cyclopentyl-ethyl)pyrazol-4-yl]pyrrolo[2,3-d]pyrimidin-7-yl]methyl 5-[(3R)-dithiolan-3-yl]pentanoate, (4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl) pentanoate, and (4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate.

[0546] [4-[(1S)-1-(2-Cyano-1-cyclopentyl-ethyl)pyrazol-4-yl]pyrrolo[2,3-d]pyrimidin-7-yl]methyl 5-[(3R)-dithiolan-3-yl]pentanoate as a beige solid (11.7 mg, 11%). LCMS (Method C): Rt=5.13 min; [M+H].sup.+=525.1. LCMS (Method D): Rt=5.17 min; [M+H].sup.+=525.1.

[0547] (4-(1-((R)-2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-2-oxido-1,2-dithiolan-3-yl) pentanoate and (4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl 5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoate as a white solid (5.2 mg, 5%). LCMS (Method C): Rt=5.75 min; [M+H].sup.+=541.1. LCMS (Method D): Rt=6.22 min; [M+H].sup.+=541.1.

Chemical Synthesis Examples 43 & 44

(3R)-3-Cyclopentyl-3-(4-(7-(5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile (43) and (3R)-3-Cyclopentyl-3-(4-(7-(5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile (44)

##STR00202##

[0548] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (63 mg, 0.33 mmol) was dissolved in anhydrous DCM (4.0 mL), 5-[(3R)-2-oxido-1,2-dithiolan-3-yl]pentanoic acid (29.1 mg, 0.13 mmol) was added and the reaction mixture stirred at r.t. for 1 h. Separately, DMAP (16 mg, 0.13 mmol) was dissolved in anhydrous DCM (4.0 mL), Ruxolitinib (40 mg, 0.13 mmol) was added and the reaction mixture stirred at r.t. for 10 min. The two solutions were combined, and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by preparative reversed-phase HPLC to afford (3R)-3-cyclopentyl-3-(4-(7-(5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile and (3R)-3-cyclopentyl-3-(4-(7-(5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile as a white solid (3.6 mg, 6%). LCMS (Method C): Rt=4.99 min; [M+H].sup.+=511.2. LCMS (Method D): Rt=5.02 min; [M+H].sup.+=511.2.

Chemical Synthesis Example 45

Methyl (R)-4-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate

##STR00203##

[0549] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (63 mg, 0.33 mmol) was dissolved in anhydrous DCM (2 mL), mono-methyl hydrogen succinate (17 mg, 0.13 mmol) was added and the reaction mixture stirred at r.t. for 1 h. Separately, DMAP (16 mg, 0.13 mmol) was dissolved in anhydrous DCM (2 mL), Ruxolitinib (40 mg, 0.13 mmol) was added and the reaction mixture was stirred at r.t. for 10 min. The two solutions were combined, and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 70% EtOAc in isohexane, to afford methyl (R)-4-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate as a white solid (43.0 mg, 78%). LCMS (Method C): Rt=4.89 min; [M+H].sup.+=421.3. LCMS (Method D): Rt=4.90 min; [M+H].sup.+=421.3.

Chemical Synthesis Example 46

Ethyl (R)-4-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimi-din-7-yl)-4-oxobutanoate

##STR00204##

[0550] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (63 mg, 0.33 mmol) was dissolved in anhydrous DCM (2.0 mL), mono-ethyl succinate (19 mg, 0.13 mmol) was then added and the mixture was stirred at r.t. for 1 h. Separately, DMAP (16 mg, 0.13 mmol) was dissolved in anhydrous DCM (2 mL), Ruxolitinib (40 mg, 0.13 mmol) was added and the reaction mixture was stirred at r.t. for 10 min. The two solutions were combined, and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 70% EtOAc in isohexane to afford ethyl (R)-4-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimi-din-7-yl)-4-oxobutanoate as a white solid (26.7 mg, 47%). LCMS (Method C): Rt=5.14 min; [M+H].sup.+=435.2. LCMS (Method D): Rt=5.17 min; [M+H].sup.+=435.2.

Chemical Synthesis Example 47

tert-Butyl (R)-4-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate

##STR00205##

[0551] 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (125 mg, 0.65 mmol) was dissolved in anhydrous DCM (3.0 mL), 4-(tert-butoxy)-4-oxobutanoic acid (46 mg, 0.26 mmol) was added and the mixture was stirred at r.t. for 1 h. Separately, DMAP (32 mg, 0.26 mmol) was dissolved in anhydrous DCM (3.0 mL), Ruxolitinib (80 mg, 0.26 mmol) was added and the mixture stirred at r.t. for 10 min. The two solutions were combined, and the reaction mixture stirred at r.t. for 16 h. The solvent was evaporated in vacuo and the crude product was purified by flash chromatography (Biotage Isolera four; 10 g Sfar cartridge) eluting with 70% EtOAc in isohexane, to afford tert-butyl (R)-4-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-oxobutanoate as a white solid (84.0 mg, 70%). LCMS (Method C): Rt=5.59 min; [M+H].sup.+=483.3. LCMS (Method D): Rt=5.62 min; [M+H].sup.+=483.3

Chemical Synthesis Examples 48 & 49

1-(4-(1-((R)-2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isobutyrate (48) and (R)-3-cyclopentyl-3-(4-(7-isobutyryl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile (49)

##STR00206##

[0552] To a solution of Ruxolitinib (75 mg, 0.23 mmol) in DMF (2.5 mL), sodium hydride (60% dispersion in oil, 10 mg, 0.42 mmol) was added and the reaction mixture stirred at r.t. for 15 min. 1-Chloroethyl 2-methylpropanoate (120 mg, 0.80 mmol) and NaI (10 mg, 0.29 mmol) were added and the reaction mixture stirred at r.t for 3 h. The reaction mixture was partitioned between water and EtOAc. The layers were separated and the organic phase washed successively with sat. NaHCO.sub.3(aq) and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford 1-(4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isobutyrate and (R)-3-cyclopentyl-3-(4-(7-isobutyryl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile.

[0553] 1-(4-(1-((R)-2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isobutyrate as a colorless gum (17.8 mg, 19%). LCMS (Method C): Rt=4.69 min; [M+H].sup.+=421.3. LCMS (Method D): Rt=4.79 min; [M+H].sup.+=421.3. .sup.1H-NMR (400 MHz, DMSO-D6) 8.79 (s, 1H), 8.71 (s, 1H), 8.35 (s, 1H), 7.89 (d, J=3.7 Hz, 1H), 7.25 (q, J=6.3 Hz, 1H), 7.09 (d, J=3.7 Hz, 1H), 4.50 (td, J=9.6, 4.1 Hz, 1H), 3.10-3.26 (m, 2H), 2.31-2.44 (m, 1H), 1.72-1.86 (m, 4H), 1.08-1.67 (m, 7H), 1.01 (d, J=6.9 Hz, 3H), 0.95 (d, J=7.3 Hz, 3H).

[0554] (R)-3-Cyclopentyl-3-(4-(7-isobutyryl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile as a white solid (20.6 mg, 24%). LCMS (Method C): Rt=5.13 min; [M+H].sup.+=377.2. LCMS (Method D): Rt=5.02 min; [M+H].sup.+=377.2.

Chemical Synthesis Examples 50 & 51

1-(4-(1-((R)-2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isopropyl carbonate (50) and isopropyl (R)-4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (51)

##STR00207##

[0555] To a solution of Ruxolitinib (50 mg, 0.16 mmol) in DMF (1.5 mL), sodium hydride (60% dispersion in oil, 7 mg, 0.29 mmol) was added and the reaction mixture stirred at r.t. for 15 min. 1-Chloroethyl isopropyl carbonate (60 mg, 0.36 mmol) and NaI (6 mg, 40 mol) were added and the reaction mixture stirred at r.t. for 4 h. The reaction mixture was partitioned between water and EtOAc. The layers were separated, and the organic phase washed successively with sat. NaHCO.sub.3(aq) and sat. brine solution, dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford 1-(4-(1-((R)-2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isopropyl carbonate and isopropyl (R)-4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate.

[0556] 1-(4-(1-((R)-2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethyl isopropyl carbonate was afforded as a white solid (15.9 mg, 22%). LCMS (Method G): Rt=7.24 min; [M+H].sup.+=437.3. LCMS (Method D): Rt=5.71 min; [M+H].sup.+=437.3.

[0557] Isopropyl (R)-4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimi-dine-7-carboxylate was afforded as a white solid (10.5 mg, 16%). LCMS (Method G): Rt=7.04 min; [M+H].sup.+=393.3. LCMS (Method D): Rt=6.48 min; [M+H].sup.+=393.3.

Chemical Synthesis Example 52

1-((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)ethyl (1r,4R)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate

##STR00208##

[0558] To a solution of Cilomilast (25.0 mg, 70.0 mol) in DMF (1.5 mL) was added DIPEA (44 L, 0.255 mmol) and 1-chloroethyl 5-[(3R)-dithiolan-3-yl]pentanoate (125 mg, 0.47 mmol) and the reaction mixture stirred at 50 C. for 48 h. To the reaction mixture was added water (3 mL) and EtOAc (3 mL) and the layers were separated. The aqueous phase was acidified and ethyl acetate (5 mL)added. The combined organic phases were dried (MgSO.sub.4) and the solvent evaporated in vacuo. The resulting residue was dissolved in DMSO and purified by preparative HPLC to afford 1-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)ethyl (1r,4R)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate (3.1 mg, 3%) as a yellow oil. LCMS (Method C): Rt =6.13 min; [M+NH.sub.4].sup.+=593.2.

Chemical Synthesis Examples 53 & 54

[0559] 1-((5-((3S)-1-Oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)ethyl (1r,4R)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate (53) and 1-((5-((3S)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)ethyl (1r,4R)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate (54)

##STR00209##

[0560] To a solution of Cilomilast (25.0 mg, 70.0 mol) in DMF (1.5 mL) was added DIPEA (44.0 L, 0.255 mmol) and 1-chloroethyl 5-[(3R)-dithiolan-3-yl]pentanoate (125 mg, 0.470 mmol). The reaction mixture was stirred at 50 C. for 48 h. To the reaction mixture was added water (3 mL) and ethyl acetate (3 mL). The layers were separated and the organic phase washed with sat. NaHCO.sub.3(aq), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by flash chromatography eluting with 20% EtOAc-isohexane.fwdarw.80% EtOAc-isohexane. The combined fractions were concentrated in vacuo to afford an orange oil. The oil was left to stand at r.t. under a blanket flow of air for 24 h to afford 1-((5-((3S)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)ethyl (1r,4S)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate and 1-((5-((3S)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)ethyl (1r,4S)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate (3.7 mg, 3%) as a yellow oil. LCMS (Method C): Rt=5.40 min; [M+H].sup.+=592.2.

Chemical Synthesis Example 55

1-(Isobutyryloxy)ethyl (1r,4r)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate

##STR00210##

[0561] To a solution of Cilomilast (25.0 mg, 70.0 mol) in DMF (1.5 mL) was added DIPEA (44.0 L, 0.255 mmol) and 1-chloroethyl isobutyrate (33.0 mg, 0.219 mmol) and the reaction mixture stirred at 50 C. for 24 h. To the reaction mixture was added water (3 mL) and ethyl acetate (3 mL). The layers were separated and the organic phase washed with NaHCO.sub.3(aq) (3 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The resulting residue was dissolved in DMSO and purified by preparative reversed-phase HPLC to afford 1-(isobutyryloxy)ethyl (1r,4r)-4-cyano-4-(3-(cyclopentyloxy)-4-methoxyphenyl)cyclohexane-1-carboxylate (4.5 mg, 14%) as an orange oil. LCMS (Method F): Rt=8.60 min; [M+NH.sub.4].sup.+=475.3.

Chemical Synthesis Example 56

(8S,9S,10R,11S,13S,14S,17R)-11,17-Dihydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid

##STR00211##

[0562] To a solution of prednisolone (0.50 g, 1.40 mmol) in THF (4.0 mL) and MeOH (1.0 mL) was added a warm solution (50 C.) of sodium periodate (0.89 g, 4.2 mmol) in water (3.3 mL). The resulting suspension was stirred at r.t. for 18 h. The reaction mixture was then concentrated in vacuo, diluted with water (5 mL) and the solid precipitate collected by filtration. The solid was further washed with water (25 mL) then dried in a vac oven at 40 C. for 16 h to afford (8S,9S,1R,11S,13S,14S,17R)-11,17-dihydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid as a white solid (0.44 g, 92%). LCMS (Method C): Rt=3.71 min; [M+H].sup.+=347.3. LCMS (Method E): Rt=2.71 min; [M+H].sup.+=347.2.

Chemical Synthesis Example 57

(8S,9S,10R,11S,13S,14S,17R)-17-((Ethoxycarbonyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid

##STR00212##

[0563] To a vigorously stirred mixture of (8S,9S,10R,11S,13S,14S,17R)-11,17-dihydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (40 mg, 0.115 mmol), DCM (3.0 mL) and water (3.0 mL) was added ethyl chloroformate (33 L, 0.35 mmol) as a solution in DCM (1.0 mL) and the mixture stirred at r.t. for 1 h. The reaction mixture was passed through a phase separator. Diethylamine (24 L, 0.23 mmol) was added and the mixture stirred at r.t. for 5 days. Additional diethylamine (24 L, 0.23 mmol) was added and the mixture stirred at r.t. for a further 4 h. The solvent was evaporated in vacuo and the crude product purified by preparative reversed-phase HPLC to afford (8S,9S,10R,11S,13S,14S,17R)-17-((ethoxycarbonyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid as a white solid (30.6 mg, 63%). LCMS (Method C): Rt=4.38 min; [M+H].sup.+=419.3. LCMS (Method E): Rt=3.05 min; [M+H].sup.+=419.3.

Chemical Synthesis Example 58

(8S,9S,10R,11S,13S,14S,17R)-17-((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid

##STR00213##

[0564] To a solution of Lipoic acid (30 mg, 0.14 mmol) in DMF (2.0 mL) was added DIPEA (60 L, 0.35 mmol) and HATU (60 mg, 0.16 mmol). The resulting solution was stirred at r.t. for 1 h. (8S,9S,10R,11S,13S,14S,17R)-11,17-Dihydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (50 mg, 0.14 mmol) was added and the reaction mixture stirred at r.t. for 7 days. The reaction mixture was diluted with EtOAc (30 mL), washed with H.sub.2O (315 mL) then sat. brine solution (15 mL). The organic phase was dried (MgSO.sub.4), filtered and the solvent evaporated to afford the crude material as a yellow gum. Purification by preparative reversed-phase HPLC afforded (8S,9S,10R,11S,13S,14S,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid as a white solid (39.6 mg, 51%). LCMS (Method C): Rt=5.13 min; [M+H].sup.+=535.2. LCMS (Method E): Rt=3.63 min; [M+H].sup.+=535.2.

Chemical Synthesis Examples 59, 60, & 61

Chloromethyl (8S,9S,10R,11S,13S,14S,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (59), chloromethyl (8S,9S,10R,11S,13S,14S,17R)-11-hydroxy-10,13-dimethyl-17-((5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (60), and chloromethyl (8S,9S,10R,11S,13S,14S,17R)-11-hydroxy-10,13-dimethyl-17-((5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (61)

##STR00214##

[0565] To a mixture of (8S,9S,10R,11S,13S,14S,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (60 mg, 0.112 mmol), tetrabutylammonium hydrogen sulfate (3.8 mg, 11 mol) and sodium bicarbonate (75 mg, 0.90 mmol) in 1:1 water-DCM (6 mL) was added chloromethyl chlorosulfate (34 L, 0.337 mmol). The reaction mixture was stirred vigorously at r.t. for 18 h. The reaction mixture was passed through a phase separator and the filtrate evaporated in vacuo. Purification by preparative reversed-phase HPLC afforded chloromethyl (8S,9S,10R,11S,13S,14S,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate, chloromethyl (8S,9S,10R,11S,13S,14S,17R)-11-hydroxy-10,13-dimethyl-17-((5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate, and chloromethyl (8S,9S,10R,11S,13S,14S,17R)-11-hydroxy-10,13-dimethyl-17-((5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate.

[0566] Chloromethyl (8S,9S,10R,11S,13S,14S,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate as an off-white solid (25.6 mg, 39%). LCMS (Method C): Rt=5.83 min; [M+H].sup.+=583.2. LCMS (Method E): Rt=5.81 min; [M+H].sup.+=583.2.

[0567] Chloromethyl (8S,9S,10R,11S,13S,14S,17R)-11-hydroxy-10,13-dimethyl-17-((5-((3R)-2-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate and chloromethyl (8S,9S,10R,11S,13S,14S,17R)-11-hydroxy-10,13-dimethyl-17-((5-((3R)-1-oxido-1,2-dithiolan-3-yl)pentanoyl)oxy)-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate as a white solid (9.5 mg, 14%). LCMS (Method C): Rt=4.98 min; [M+H].sup.+=599.2. LCMS (Method E): Rt=4.98 min; [M+H].sup.+=599.2.

Chemical Synthesis Example 62

(R)-2,2-Dimethylthiazolidine-4-carboxylic acid

##STR00215##

[0568] A suspension of L-cysteine (5.00 g, 40.0 mmol) in anhydrous acetone (125 mL) was heated under reflux (80 C.) under an atmosphere of nitrogen for 40 h. The reaction mixture was cooled to r.t., filtered through a Celite cartridge, concentrated to 50% of its original volume then left to stand at r.t for 24 h. The solid was collected, washed with acetone (10 mL) then dried to afford (R)-2,2-dimethylthiazolidine-4-carboxylic acid as a white solid (5.29 g, 82%). .sup.1H-NMR (400 MHz, D.sub.2O) 4.41-4.45 (m, 1H), 3.46 (dd, J=12.4, 8.2 Hz, 1H), 3.31 (dd, J=12.4, 7.3 Hz, 1H), 1.66 (s, 3H), 1.64 (s, 3H).

Chemical Synthesis Example 63

(R)-3-Acetyl-2,2-dimethylthiazolidine-4-carboxylic acid

##STR00216##

[0569] To a mixture of (R)-2,2-dimethylthiazolidine-4-carboxylic acid (2.00 g, 12.4 mmol) in acetone (100 mL) under an atmosphere of N.sub.2 was added acetic anhydride (2.30 mL, 24.8 mmol) followed by DBU (3.70 mL, 24.8 mmol). The resultant solution was stirred at r.t. for 18 h. Sat. aqueous NH.sub.4Cl (30 mL) was added to the reaction mixture and then stirred for 10 min. The mixture was extracted into EtOAc (250 mL), washed with water (50 mL) then sat. brine solution (50 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo to afford a first batch of (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylic acid as a brown solid (0.82 g, 33%). LCMS (Method A): Rt=1.15 min; [MH].sup.=202.1. LCMS (Method B): Rt=0.51 min; [MH].sup.=202.1.

[0570] 2M HCl.sub.(aq) was added to the combined aqueous washings until pH 1 was reached. The product was extracted into EtOAc (250 mL). The combined organics were washed with sat. brine solution (50 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo to afford a second batch of (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylic acid as a yellow solid (1.56 g, 62%). LCMS (Method A): Rt=1.12 min; [MH].sup.=202.1. LCMS (Method B): Rt=0.29 min; [MH].sup.=202.1. .sup.1H-NMR (400 MHz, ACETONE-D6) 11.17 (br s, 1H), 5.09 (dd, J=6.0, 1.4 Hz, 1H), 3.41 (dd, J=11.9, 6.0 Hz, 1H), 3.30 (dd, J=11.9, 0.9 Hz, 1H), 2.03 (s, 3H), 1.83 (s, 3H), 1.79 (s, 3H).

Chemical Synthesis Example 64

(8S,9S,10R,11S,13S,14S,17R)-17-(((R)-3-Acetyl-2,2-dimethylthiazolidine-4-carbonyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid

##STR00217##

[0571] To a solution of (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylic acid (29 mg, 0.144 mmol) in DMF (2.0 mL) was added DIPEA (0.060 mL, 0.346 mmol) and HATU (60 mg, 0.159 mmol). The resulting solution was stirred at r.t. for 1 h. (8S,9S,10R,11S,13S,14S,17R)-11,17-Dihydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (50 mg, 0.144 mmol) was added to the reaction mixture and stirring continued at r.t. for 16 h. The reaction mixture was diluted with EtOAc (30 mL), washed with water (315 mL) then sat. brine solution (15 mL), dried (MgSO.sub.4), filtered and the solvent evaporated in vacuo. The crude product was purified by preparative reversed-phase HPLC to afford (8S,9S,10R,11S,13S,14S,17R)-17-(((R)-3-acetyl-2,2-dimethylthiazolidine-4-carbonyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid as a white solid (27.2 mg, 35%). LCMS (Method A): Rt=1.64 min; [M+H].sup.+=532.2.

Chemical Synthesis Example 65

(8S,9S,10R,11S,13S,14S,17R)-17-((Acetyl-L-cysteinyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid

##STR00218##

[0572] A solution of (8S,9S,10R,11S,13S,14S,17R)-17-(((R)-3-acetyl-2,2-dimethylthiazolidine-4-carbonyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (58 mg, 0.11 mmol) in trifluoroacetic acid (2.0 mL) was heated at 40 C. for 3 h. The reaction mixture was evaporated to dryness in vacuo then purified by preparative reversed-phase HPLC to afford (8S,9S,10R,11S,13S,14S,17R)-17-((acetyl-L-cysteinyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (34.0 mg, 63%) as a white solid. LCMS (Method C): Rt=3.90 min; [M+Na]=514.2. LCMS (Method E): Rt=2.91 min; [M+H].sup.+=492.1.

Chemical Synthesis Example 66

(8S,9S,10R,11S,13S,14S,17R)-17-((Chloromethoxy)carbonyl)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylate

##STR00219##

[0573] To a mixture of (8S,9S,10R,11S,13S,14S,17R)-17-(((R)-3-acetyl-2,2-dimethylthiazolidine-4-carbonyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (50 mg, 0.094 mmol), tetrabutylammonium hydrogen sulfate (3.2 mg, 0.009 mmol) and sodium bicarbonate (63 mg, 0.75 mmol) in 1:1 DCM-H.sub.2O (6 mL) was added chloromethyl chlorosulfate (29 L, 0.28 mmol). The reaction mixture was stirred vigorously at r.t. for 18 h. The reaction mixture was passed through a phase separator with additional washing with DCM (3 mL) then the combined filtrate evaporated in vacuo to afford (8S,9S,10R,11S,13S,14S,17R)-17-((chloromethoxy)carbonyl)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylate as a yellow gum (53.4 mg, 98%). This was used without further purification. LCMS (Method A): Rt=1.84 min; [M+H].sup.+=580.2.

Chemical Synthesis Example 67

Chloromethyl (8S,9S,10R,11S,13S,14S,17R)-17-((acetyl-L-cysteinyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate

##STR00220##

[0574] A solution of (8S,9S,10R,11S,13S,14S,17R)-17-((chloromethoxy)carbonyl)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylate (53.4 mg, 90 mol) in trifluoroacetic acid (2.0 mL) was heated at 40 C. for 3 hours. The reaction mixture was evaporated to dryness in vacuo, redissolved in DMSO then purified by preparative reversed-phase HPLC to afford chloromethyl (8S,9S,10R,11S,13S,14S,17R)-17-((acetyl-L-cysteinyl)oxy)-11-hydroxy-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (23.6 mg, 47%) as a white solid. LCMS (Method C): Rt=4.58 min; [M+Na]=562.1. LCMS (Method E): Rt=4.58 min; [M+H].sup.+=540.2.

Chemical Synthesis Example 68

(6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid

##STR00221##

[0575] To a solution of (6S,8S,9R,10S,11S,13S,14S,16R,17R)-6,9-difluoro-11,17-dihydroxy-10,13,16-trimethyl-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (100 mg, 0.252 mmol) in DMF (2.0 mL) was added (R)-(+)-Lipoic acid (52 mg, 0.252 mmol) and HATU (115 mg, 0.303 mmol) followed by DIPEA (0.13 mL, 0.757 mmol). The reaction was stirred for 2 hours at r.t. The product was purified directly by prep HPLC to give (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (116 mg, 0.198 mmol, 78%) as a yellow foam. LC-MS (Method H): Rt=1.88, m/z [M+H].sup.+=585.3; [M+Na].sup.+=607.4, 100%.

Chemical Synthesis Example 69

Cyanomethyl (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (73A)

##STR00222##

[0576] To a solution of (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((5-((R)-1,2-Dithiolan-3-yl)pentanoyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (116 mg, 0.198 mmol) in DMF (2.0 mL) was added chloroacetonitrile (13 L, 0.198 mmol) and TEA (55 L, 0.395 mmol) and the reaction was stirred for 72 hours. The reaction was concentrated in vacuo and the residue was purified by column chromatography (silica, sfar 10 g, 0-10% MeOH in DCM) to give cyanomethyl (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((5-((R)-1,2-dithiolan-3-yl)pentanoyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (101 mg, 0.161 mmol, 81%) as an off-white solid. LC-MS (Method J): Rt=5.55, [M+H].sup.+=624.4, 99%, LC-MS (Method K): Rt=5.53, [M+H].sup.+=624.3, 99%. .sup.1H NMR (400 MHz, DMSO-d6) 7.22-7.26 (m, 2H), 7.15-7.17 (m, 1H), 6.29 (dd, J=10.1, 1.8 Hz, 1H), 6.10 (s, 1H), 5.53-5.70 (m, 2H), 4.98 (s, 2H), 4.18 (t, J=4.1 Hz, 1H), 3.53-3.59 (m, 1H), 3.05-3.21 (m, 2H), 2.31-2.40 (m, 3H), 2.21-2.29 (m, 2H), 2.02-2.12 (m, 2H), 1.78-1.87 (m, 2H), 1.59-1.66 (m, 2H), 1.48-1.56 (m, 6.5H), 1.32-1.40 (m, 1.5H), 1.19-1.25 (m, 1H), 0.98 (s, 3H), 0.85 (d, J=7.3 Hz, 3H).

Chemical Synthesis Example 70

(6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-(((R)-3-Acetyl-2,2-dimethylthiazolidine-4-carbonyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid

##STR00223##

[0577] To a solution of (4R)-3-acetyl-2,2-dimethyl-thiazolidine-4-carboxylic acid (77 mg, 0.378 mmol) in DCM (5.0 mL) was added (6S,8S,9R,10S,11S,13S,14S,16R,17R)-6,9-difluoro-11,17-dihydroxy-10,13,16-trimethyl-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (150 mg, 0.378 mmol), HATU (173 mg, 0.454 mmol) and DIPEA (0.20 mL, 1.14 mmol). The solution was left to stir at r.t. for 3 hours. The reaction was concentrated in vacuo and purified by prep HPLC to give (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-(((R)-3-acetyl-2,2-dimethylthiazolidine-4-carbonyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (168 mg, 0.289 mmol, 76%) as a white solid. LC-MS (Method H): Rt=1.77, m/z [M+H].sup.+=582.2; 95%.

Chemical Synthesis Example 71

(6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((Cyanomethoxycarbonyl)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-yl (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylate

##STR00224##

[0578] To a solution of (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-(((R)-3-acetyl-2,2-dimethylthiazolidine-4-carbonyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylic acid (168 mg, 0.289 mmol) in DMF (1.0 mL) was added chloroacetonitrile (22 uL, 0.347 mmol) and TEA (81 uL, 0.578 mmol). The reaction was left to stir at r.t. for 16 hours. The solvent was removed in vacuo and redissolved in DCM (10 mL) before being washed with sat. NaHCO.sub.3 (10 mL) followed by sat. NH.sub.4Cl (10 mL) and brine (10 mL) before being passed through a phase separator and concentrated in vacuo. The crude was purified by flash column chromatography (silica, sfar 10 g, 0-10% MeOH in DCM) to give (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((cyanomethoxycarbonyl)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-yl (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylate (37 mg, 0.0596 mmol, 20%,) as an off-white solid. LC-MS (Method L): Rt=2.73, m/z 621.3 M+H].sup.+, 84%.

Chemical Synthesis Example 72

Cyanomethyl (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((acetyl-L-cysteinyl)oxy)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (73B)

##STR00225##

[0579] To a solution of (6S,8S,9R,10S,115,13S,14S,16R,17R)-17-((cyanomethoxycarbonyl)-6,9-difluoro-11-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-yl (R)-3-acetyl-2,2-dimethylthiazolidine-4-carboxylate (37 mg, 0.0596 mmol) in DCM (0.20 mL) in a sealed vial was added TFA (0.80 mL). The reaction was heated to 40 C. and left to stir for 5 hours. The reaction was concentrated in vacuo and purified by prep HPLC to give cyanomethyl (6S,8S,9R,10S,11S,13S,14S,16R,17R)-17-((acetyl-L-cysteinyl)oxy)-6,9-difluoro-11-hydroxy--10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate (20 mg, 0.0344 mmol, 58%) as a white solid. LC-MS (Method J): Rt=4.38 min; [M+Na].sup.+=603.4, 92%, LC-MS (Method K): Rt=4.36 min; [M+H].sup.+=581.4, 84%. .sup.1H NMR (400 MHz, DMSO-d6) 8.27 (d, J=7.8 Hz, 1H), 7.26 (dd, J=10.1, 1.4 Hz, 1H), 6.30 (dd, J=10.5, 1.8 Hz, 1H), 6.11 (s, 1H), 5.71-5.54 (m, 2H), 4.95-4.97 (m, 2H), 4.40 (td, J=7.9, 4.7 Hz, 1H), 4.18 (td, J=4.6, 2.0 Hz, 1H), 3.13-3.18 (m, 1H), 2.78-2.85 (m, 1H), 2.65-2.75 (m, 1H), 2.59 (t, J=8.7 Hz, 1H), 2.24 (dd, J=6.6, 4.4 Hz, 1H), 2.00-2.12 (m, 2H), 1.76-1.85 (m, 4H), 1.60 (d, J=13.3 Hz, 1H), 1.43-1.52 (m, 4H), 1.14-1.20 (m, 1H), 0.98 (s, 3H), 0.87 (d, J=7.3 Hz, 3H).

[0580] Other compounds provided herein (e.g., in Table 1-9) are prepared according to a similar process as provided for the examples provided hereinabove (e.g., Chemical Synthesis Examples 1-72).

II. Biological Evaluation

Example 1: Rabbit Cornea Homogenate Stability Assay

[0581] Determining Rabbit Cornea Homogenate stability of the test compounds was performed using UPLC-MS. The assay was performed at two concentrations of Rabbit Cornea Homogenate (0.15 mg/mL and 0.45 mg/mL total protein) so that any hydrolysis observed can be assigned as esterase dependent or not.

Rabbit Cornea Homogenisation

[0582] Three to four rabbit corneas (e.g., New Zealand Whites (NZW) or Dutch Belted (DB)) of approx. 50 mg each were sliced and scraped with a scalpel and tweezers until reduced to small (1-3 mm), thin pieces. These were transferred into a glass vial containing approximately 2 mL of cold DPBS pH 7.4 buffer.

[0583] Sample was cooled intermittently on ice and shear homogenized for 3 minutes, then centrifuged for 3 min at 13,000 g. The supernatant was pipetted off into a vial, and total protein concentration determined at 280 nm. Sample was stored at 78 C.

Rabbit Cornea Esterase Assay

Preparation of Stock Solutions:

[0584] 10 mM Compound DMSO stocks were diluted to 10 M in a glass vial: 10 L of 10 mM Compound stock was added to 9,990 l 50 mM DPBS, pH 7.4 buffer. Esterase homogenate was diluted to 300 ng/l and 900 ng/L in DPBS.

Assay Conditions:

[0585] A heater shaker was set to 37 C. Into a suitable 96 well plate (Run Plate), 70 L of 300 or 900 ng/l esterase homogenate was pipetted into two rows as compounds were analyzed in duplicate (2 min, 5 min, 10 min, 20 min and 45 min). The plate was sealed and then warmed at 37 C. for 5 min.

[0586] Two 96 deep-well plates were put on ice (Kill Plates). To these, 990 L of 50:50 MeCNH.sub.2O were added to required rows, labelled 0 min 2 min, 5 min, 10 min, 20 min and 45 min. The plates were covered to minimize evaporation.

[0587] To both rows of the Run Plate, 70 L of 10 M compound solution was added. At the appropriate time point, 10 L of the assay mixture was added to the matching kill plate well containing 990 L of 50:50 cold MeCNH.sub.2O. Samples were analyzed as soon as practicable by UPLC-MS (Waters Xevo TQ-S).

Assay Conditions for Lipoic Acid Analysis:

[0588] A heater shaker was set to 37 C. Into a suitable 96 well plate (Run Plate), 80 L of 300 or 900 ng/L esterase homogenate was pipetted into two rows as compounds were analyzed in duplicate (2 min, 5 min, 10 min, 20 min and 45 min). The plate was sealed and then warmed at 37 C. for 5 min.

[0589] Two 96 shallow-well plates were placed on ice (Kill Plates). To these, 180 L of 60:40 MeCN-H.sub.2O+0.1% acetic acid are added to required rows. The plates were sealed to minimize evaporation.

[0590] To both rows of the Run Plate, 80 L of 10 M compound solution was added. At the appropriate time point, 20 L of the assay mixture was added to the matching kill plate well containing 180 L of 60:40 cold MeCN-H.sub.2O+0.1% acetic acid. For lipoic acid analysis, samples were analyzed as soon as practicable by LCMS (Waters Xevo TQ-S). For parent conjugate and parent analysis the samples are diluted further 1 in 10: 20 L supernatant is added to 180 L of 50:50 MeCNH.sub.2O.

[0591] Parent conjugate, parent and keratolytic concentrations were determined against appropriate standard response curves and the half-life (T1/2) of the parent conjugate was calculated using the measured concentration of the parent conjugate at each time point in the linear region of the log linear plot.

Example 2: Aqueous Hydrolysis Stability Assay

[0592] Determination of aqueous stability of the test compounds was performed using UPLC-MS. A test compound 10 mM stock solution was prepared in DMSO. 10 L of the DMSO stock solution was dissolved in 990 L of DPBS pH 7.4 buffer to prepare a 100 M stock. A further dilution was made by dissolving 75 l of 100 M stock into 225 L of DPBS. Final DMSO concentration is 0.25%. The solution is kept at 37 C. and injected without delay into the LCMS (Waters Xevo TQ-). Additional injections were performed at appropriate time points.

[0593] Half-life (T.sub.1/2) of the parent conjugate was calculated using the peak area or measured concentration of the parent conjugate at each time point in the linear region of the loglinear plot.

[0594] Aqueous Stability and Hydrolysis Rates of Example Compounds

TABLE-US-00012 TABLE 11 Aqueous Stability Esterase Esterase Esterase Chemical (DPBS) (0.15 mg/mL) (0.45 mg/mL) (0.90 mg/mL) Synthesis % parent released % parent released % parent released % parent released Example(s) after t min after 45 min after 45 min after 45 min 12 A (t = 44) B (NZW) B (NZW) n.t. 13 & 14 A (t = 44) A (NZW) A (NZW) n.t. 15 A (t = 44) D (NZW) D (NZW) n.t. 16 & 17 A (t = 44) A (NZW) B (NZW) n.t. 18 A (t = 44) B (NZW) B (NZW) n.t 19 A (t = 44) B (NZW) B (NZW) n.t. 21 A (t = 45) C (NZW) D (NZW) n.t. 22 n.t. D (NZW) D (NZW) n.t. 23 n.t A (NZW) A (NZW) n.t. 24 n.t. A (NZW) A (NZW) n.t. 25 A (t = 44) D (NZW) D (NZW) n.t. 26 A (t = 44) B (NZW) B (NZW) n.t. 27 A (t = 44) B (NZW) C (NZW) n.t. 28 n.t. n.t. D (DB) D (DB) 29 n.t. B (NZW) C (NZW) D (DB) D (DB) 30 B (t = 43) B (NZW) B (NZW) n.t. 31 A (t = 44) C (NZW) C (NZW) n.t. 32 & 33 n.t. B (NZW) C (NZW) n.t. 34 n.t. C (NZW) C (NZW) n.t. 35 & 36 A (t = 43) B (NZW) B (NZW) n.t. 37 A (t = 42) D (NZW) * D (NZW) * n.t. 38 A (t = 48) B (NZW) C (NZW) n.t. 39 B (t = 45) B (NZW) C (NZW) n.t. 40 A (t = 45) D (NZW) D (NZW) n.t. 41 & 42 n.t. n.t. D (DB) D (DB) 43 & 44 n.t. B (NZW) B (NZW) n.t. 45 A (t = 44.2) C (NZW) D (NZW) n.t. 46 n.t. C (NZW) D (NZW) n.t 47 A (t = 45) B (NZW) B (NZW) n.t. 48 n.t. A (NZW) A (NZW) n.t. 49 n.t. D NZW) D (NZW) n.t. 50 B (t = 44) D (DB) D (DB) n.t. 51 A (t = 45) A (DB) B (DB) n.t. 52 n.t. D (DB) D (DB) n.t. 53 & 54 A (t = 45) C (NZW) C (NZW) n.t. 55 A (t = 45) D (DB) D (DB) n.t. A: <25%; B: 25% to 50%; C: 51% to 75%; D: >75%; n.t. = not tested. * n = 2

TABLE-US-00013 TABLE 12 Aqueous Chemical Stability Esterase Esterase Esterase Esterase Synthesis (DPBS) T.sub.1/2 (0.15 mg/mL) (0.45 mg/mL) (0.9 mg/mL) (2.0 mg/mL) Example(s) after t min T.sub.1/2 (45 min) T.sub.1/2 (45 min) T.sub.1/2 (45 min) T.sub.1/2 (45 min) 58 >120 (t = 44 min) >120 (NZW) >120 (NZW) n.t. n.t. 59 31.6 (t = 45) n.t. n.t. 22.6 (NZW) 21.3 (NZW) 60 & 61 n.t. 117 (NZW) 98 (NZW) n.t. n.t. 65 33.7 (t = 42) >120 (NZW) 119 (NZW) n.t. n.t. 67 >120 (t = 44) n.t. n.t. 47.7 (NZW) 33.7 (NZW) a: <40 min; b: 40 min to 80 min; c: >80 min; n.t. = not tested.

TABLE-US-00014 TABLE 13 Aqueous Stability Esterase Esterase Esterase Chemical (DPBS) (0.15 mg/mL) (0.45 mg/mL) (0.90 mg/mL) Synthesis % parent released % parent released % parent released % parent released Example(s) after t min after 45 min after 45 min after 45 min 73A A (t = 45) A A n.t. 73B A (t = 45) A A n.t. A: <25%; B: 25% to 50%; C: 51% to 75%; D: >75%; n.t. = not tested.

Example 3: Mouse Model of Experimental Dry Eye Disease

[0595] Female C57BL/6 mice (6-8 weeks old) or female HEL BCR Tg mice (6-8 weeks old) are commercially obtained. Experimental dry eye is induced as described by Niederkorn, et al. (J. Immunol. 2006,176:3950-3957) and Dursun et al. (Invest. Ophthalmol. Vis. Sci. 2002, 43:632-638). In brief, mice are exposed to desiccating stress in perforated cages with constant airflow from fans positioned on both sides and room humidity maintained at 30% to 35%. Injection of scopolamine hydrobromide (0.5 mg/0.2 mL; Sigma-Aldrich, St. Louis, MO) is administered subcutaneously, three times a day (8:00 AM, 12:00 noon, and 5:00 PM), on alternating hind-flanks to augment disease. Mice are exposed to desiccating stress for 3 weeks. Untreated control mice are maintained in a nonstressed environment at 50% to 75% relative humidity without exposure to forced air. Test animals are exposed to test compound and subsequently tear samples are obtained to determine stability of test compounds, and tissue samples are taken to determine presence of pro-inflammatory biomarkers.

Example 4: Thiol Assay

Stratum Corneum Preparation

[0596] Epidermis pieces are transferred and incubated overnight from 25-37 C. in a container containing 100 mL of 0.0005% trypsin (diluted in PBS). The stratum corneum pieces are removed and washed twice with HPLC grade water in a petri dish (145 mm), removing intact cells. The dish and/or pieces are shaken, producing nearly transparent layers. The stratum corneum is then transferred to a petri dish (145 mm), washed with hexane, and shaken to remove fats. Each piece is gently mounted on an absorbent paper. Each piece is transferred to an Eppendorf tube, allowing residual solvents to evaporate for a few minutes.

Thiol Assay

[0597] Compounds (e.g., 50 L; 1 M to 800 M) are applied to the isolated stratum corneum at room temperature for a period of 1-24 hours. The pieces are gently mixed with the compounds by pipetting. Following incubation, about 200 L of 10 M sodium hydroxide is added, incubating for 1 hour at room temperature with continuous blending (e.g., vortexing) until the stratum corneum disintegrates. About 200 L of 10 M hydrochloric acid is added to normalize pH, vortexing the samples. The samples are centrifuged (e.g., for 20 min at 16,000g) at room temperature. The supernatant (middle layer) is transferred to an Eppendorf tube. The free thiols are isolated by adding tricholoracetic acid (e.g., 400 L) and vortexing. The tubes are centrifuged (e.g., for 10 min at 16,000g) at room temperature. The supernatant is removed, and Ellman's reagent solution (e.g., 220 L) is added to the remaining pellet. After mixing, 100 L for each tube is transferred to a 96 well plate in the dark. The plate is incubated for about 5 minutes at room temperature while shaking. The optical absorbance at 412 nm is detected and recorded.

Example 5: Glucocorticoid Binding Assay

[0598] GR (h) (agonist radioligand) assay performed by Eurofins Cerep, France using a method similar to the one described hereinbelow.

[0599] The human lymphoblast cell line IM9 is used as a source of the soluble glucocorticoid receptor (GR). The cells are grown to densities of 1 to 1010.sup.5 cells per milliliter in RPMI 1640 media containing 10% fetal bovine serum, penicillin (100 U/ml), streptomycin (100 g/ml), and 2 mM L-glutamine at 370 and 7% CO2 in a humidified incubator. The IM9 cells are harvested from the media by centrifugation for 10 minutes at 1500 g. Cells are washed in 12 volumes of Dulbecco's phosphate-buffered saline (PBS) and re-pelleted. Washed cells are resuspended in five to six volumes (per volume of packed cells) of homogenization buffer (10 mM TES, 10 mM sodium molybdate, 1 mM EDTA, pH 7.4, 20 mM 2-mercaptoethanol, and 10% glycerol), and the cells are broken by nitrogen cavitation using 215 minutes at 600 to 750 psi nitrogen in the N.sub.2 cavitator at 0 C. Cell disruption is confirmed by Hoffman contrast microscopy using a Nikon Diaphot. The broken cell preparation is then centrifuged at 27,000 g for 15 minutes, and the resultant supernatant was centrifuged at 103,000 g for 60 minutes at 4 C. The amount of protein in the supernatant fraction is determined using a BCA assay kit with a bovine serum albumin standard. Aliquots of the supernatant fraction are snap frozen in a dry ice-acetone bath and stored at 70 C. Competitive binding assays are done in duplicate in homogenization buffer (total volume of 200 IA) by mixing 1 mg of IM9 cytosol, 0.05/Ci (3 nM) of .sup.3H-dexamethasone, and Compounds described herein (10.sup.5 to 10.sup.11 M). After incubation at 0 C. for 16 to 18 hours, the assay is stopped by the addition of 100 g of a charcoal-dextran mixture (2% activated charcoal, 0.5% dextran in 10 mM Tris, 1 mM EDTA, pH 7.4). The assay mixture is further incubated at 0 C. for 10 minutes before being centrifuged for 5 minutes at 8200 g. A 100-l sample of the supernatant (protein-bound steroid fraction) is assayed for radioactivity by liquid scintillation spectrometry, and the IC.sub.50 values were determined graphically.

[0600] In some instances, the glucocorticoid binding of a compound described herein is provided in Table 14. In some instances, a free-acid is inactive in the glucocorticoid binding assay. In some instances, an ester is active in the glucocorticoid binding assay. In some instances, an ester retains activity, such as when attached to a keratolytic agent (or a radical thereof), in the glucocorticoid binding assay.

TABLE-US-00015 TABLE 14 Chemical Synthesis Glucocorticoid Example(s) EC.sub.50 65 0.35 nM 69 >100 nM 68 21 nM 70 >100 nM

III. Preparation of Pharmaceutical Dosage Forms

Example 1: Solution for Topical Ophthalmic Use

[0601] The active ingredient is a compound of any one of Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9, or a pharmaceutically acceptable salt thereof, and is formulated as a solution with a concentration of from 0.1-1.5% w/v.