Poly inflammatory kinase inhibitors and pharmaceutical compositions

Abstract

Disclosed herein are poly-inflammatory kinase (PINK) inhibitors, compositions containing PINK inhibitors. Methods for treating malignant diseases and autoimmune disorders using the PINK inhibitors are also disclosed.

Description

[0001] POLY INFLAMMATORY KINASE INHIBITORS AND PHARMACEUTICAL COMPOSITIONS

[0002] CROSS-REFERENCE TO RELATED APPLICATION

[0003] This application claims priority to U.S. Provisional Patent Application No. 63 / 636,377 filed on April 19, 2024, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0004] FIELD

[0005] The present disclosure relates to poly INflammatory Kinase (PINK) inhibitors, pharmaceutical compositions that include the poly INflammatory Kinase inhibitors, and treatment methods including the administration of the poly inflammatory inhibitors.

[0006] BACKGROUND

[0007] Psoriasis and atopic dermatitis (AD), known as eczema, are the two most common chronic, noncommunicating inflammatory skin diseases. The prevalence of AD is 10-30% in children, and 2-10% in adults, while psoriasis affects about 3% of population. Due to the function of the skin as the primary line of defense against external pathogens, both diseases are associated with significant comorbidity. The etiology and pathogenesis of both diseases is complex and not completely understood. Though it has been shown that immune dysregulation is an important component of skin inflammation, many other factors contribute to the progression of both diseases.

[0008] Due to the poorly understood pathogenesis, treatment of both psoriasis and AD is challenging and there is no cure for any of these diseases currently. Two general approaches to treatment of these diseases have involved either topical treatment or systemic therapy. The most common topical treatment for psoriasis includes administering corticosteroids and vitamin D analogs (e.g., calcipotriol and tacalcitol), which unfortunately have limited success. Recently, a more specific topical treatment for psoriasis ( / .e., Tapinarof), which targets the aryl hydrocarbon receptor cream has been developed, although the high cost of such treatment renders this option not readily accessible to everyone. The topical treatments for AD include mostly corticosteroids and, to the lesser extent, topical calcineurin inhibitors, both of which also have limited success. Systemic treatment for both diseases includes administering corticosteroids and systemic immunomodulators (for psoriasis and psoriatic arthritis), which include TNFalpha inhibitors, IL-17alpha inhibitors, IL-17 receptor inhibitor, IL-12 and IL-23 inhibitors.

[0009] Adherence to treatments, side effects of systemic administration (e.g. generalized immunosuppression), side effects of topical administration, restrictions regarding duration and sites of application, limited therapeutic potency and frequent relapse are all recognized problems with current AD and psoriasis treatments. Thus, there is an unmet need for the novel treatment for both psoriasis and AD.

[0010] Thus, to address the foregoing issues with the treatment of AD and psoriasis, provided herein are compounds and compositions that are capable of inhibiting multiple pro-inflammatory kinases that have a role in AD and psoriasis symptom induction and / or exacerbation. Methods that apply these compounds and compositions for the treatment of AD and psoriasis are also provided herein.

[0011] SUMMARY

[0012] One aspect of the present disclosure is a compound of Formula I:

[0013]

[0014] wherein:

[0015] R1is selected from any naturally occurring amino acid side group, an alkyl group, a cycloalkyl group, a spirocyclic group, an aryl group, an haloalkyl group, a heteroaryl group, an alkyloxy group, ora thioalkyl group; and

[0016] R2is selected from -H, an alkyl group, or one or more amino acids, or a pharmaceutically acceptable salt or stereoisomer thereof. Another aspect of the present disclosure is a topical formulation including i.e., comprising) a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof; and one or more cosmetic or pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic vehicles, water soluble vehicles, emulsifiers, buffers, humectants, moisturizers, solubilizers, preservatives, colorants, plasticizers, carriers, nanocarriers, nanosuspensions, nanocrystals, and / or excipients.

[0017] Another aspect of the present disclose is a method for treating a skin condition in a subject in need thereof that includes administering a topical composition containing a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or stereoisomer thereof, to the subject in need.

[0018] Another aspect of the present disclosure is a method for treating a malignant disease and / or an autoimmune disorder in a subject in need thereof that includes administering a composition containing a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or stereoisomer thereof to the subject in need.

[0019] BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Other features and advantages of the compositions, devices and methods disclosed herein will be apparent to those skilled in the art reading the following detailed description in conjugation with the exemplary embodiments illustrated in the drawings, wherein:

[0021] FIG. 1 depicts the IMQ-induced severe psoriasis preclinical model used to examine the effect that exemplary PINK inhibitors have on kinase activities.

[0022] FIG. 2 depicts a volcano plot showing differential protein tyrosine kinase (PTK) activities after 24-hour DD-L treatment. Kinases demonstrating decreased activity after 24-hour DD-L treatment are shown in the upper left quadrant of the volcano plot.

[0023] FIG. 3 depicts a volcano plot showing differential serine / threonine kinase (STK) activity after 24-hour DD-L treatment. Kinases demonstrating decreased activity after 24-hour DD-L treatment are shown in the upper left quadrant of the volcano plot.

[0024] FIG. 4 depicts a volcano plot showing differential STK activity after 48-hour DD-L treatment. Kinases demonstrating decreased activity after 48-hour DD-L treatment are shown in the upper left quadrant of the volcano plot.

[0025] FIG. 5 depicts a volcano plot showing differential STK activity after 72-hour DD-L treatment. Kinases demonstrating decreased activity after 72-hour DD-L treatment are shown in the upper left quadrant of the volcano plot.

[0026] FIG. 6 depicts a volcano plot showing differential STK activity after 48-hour DD-I treatment. Kinases demonstrating decreased activity after 48-hour DD-I treatment are shown in the upper left quadrant of the volcano plot.

[0027] FIG. 7 depicts a volcano plot showing differential STK activity after 72-hour DD-I treatment. Kinases demonstrating decreased activity after 72-hour DD-I treatment are shown in the upper left quadrant of the volcano plot.

[0028] FIG. 8 depicts a volcano plot showing differential PTK activity after 24-hour DD-V treatment. Kinases demonstrating decreased activity after 24-hour DD-V treatment are shown in the upper left quadrant of the volcano plot.

[0029] FIG. 9 depicts a volcano plot showing differential PTK activity after 48-hour DD-V treatment. Kinases demonstrating decreased activity after 48-hour DD-V treatment are shown in the upper left quadrant of the volcano plot.

[0030] FIG. 10 depicts a volcano plot showing differential PTK activity after 72-hour DD-V treatment. Kinases demonstrating decreased activity after 72-hour DD-V treatment are shown in the upper left quadrant of the volcano plot.

[0031] FIG. 11 depicts a volcano plot showing differential STK activity after 48-hour DD-V treatment. Kinases demonstrating decreased activity after 48-hour DD-V treatment are shown in the upper left quadrant of the volcano plot.

[0032] FIG. 12 depicts a volcano plot showing differential STK activity after 72-hour DD-V treatment. Kinases demonstrating decreased activity after 72-hour DD-V treatment are shown in the upper left quadrant of the volcano plot.

[0033] FIG. 13 depicts an in vivo treatment method of severe psoriasis fortesting of therapeutic efficacy of exemplary PINK inhibitors. FIG. 14 depicts changes in epidermal thickness following IMQ-induced skin inflammation in mice for a severe psoriasis preclinical model before treatment with PINK inhibitors.

[0034] FIG. 15 depicts an in vivo treatment method of mild-to-moderate psoriasis fortesting of therapeutic efficacy of exemplary PINK inhibitors.

[0035] FIG. 16 depicts an in vivo treatment method of severe eczema for testing of therapeutic efficacy of exemplary PINK inhibitors.

[0036] FIG. 17 depicts changes in epidermal thickness following TPA-induced skin inflammation in mice for a severe eczema preclinical model before treatment with PINK inhibitors.

[0037] FIG. 18 depicts an in vivo treatment method of mild-to-moderate eczema for testing of therapeutic efficacy of exemplary PINK inhibitors.

[0038] FIG. 19 depicts H&E images of mouse ear skin cross-sections (10um cryosections). Scale bars = 20um. DMSO is the control forTPA. Cream base is the control of 5% IMQ. Vehicle refers to the vehicle of the inhibitors.

[0039] FIG. 20 depicts changes in epidermal thickness following IMQ-induced skin inflammation in mice following the treatment with DD-L, DD-I, DD-V or DMSO (control). Each experiment involved at least 5 animals in each group. *P<0.05; **P < 0.01 ; ***P<0.001 ; ****P<0.0001.

[0040] FIG. 21 depicts changes in epidermal thickness following TPA-induced skin inflammation in mice following the treatment with DD-L, DD-I, DD-V or DMSO (control). Each experiment involved at least 5 animals in each group. *P<0.05; **P < 0.01 ; ***P<0.001 ; ****P<0.0001.

[0041] FIG. 22 depicts changes in epidermal thickness following IMQ-induced skin inflammation in mice following the treatment with DD-L, DD-I, DD-V or DMSO (control). Each experiment involved at least 5 animals in each group. *P<0.05; **P < 0.01 ; ***P<0.001 ; ****P<0.0001.

[0042] FIG. 23 depicts changes in epidermal thickness following TPA-induced skin inflammation in mice following the treatment with DD-L, DD-I, DD-V, or DMSO (control). Each experiment involved at least 5 animals in each group. *P<0.05; **P < 0.01 ; ***P<0.001 ; ****P<0.0001. FIG. 24 depicts the therapeutic effect of topical treatment with PINK inhibitors and steroids on mice with IMQ-induced mild-to-moderate psoriasis.

[0043] Topical treatment with 0.1% hydrocortisone as a single compound (current standard treatment) had a therapeutic effect (middle column vs. left column).

[0044] Treatment with a mixture of DD-L and 0.1% hydrocortisone, and with DD-V and 0.1% hydrocortisone had a significantly increased therapeutic effect (p<0.01) on psoriasis, as compared to the treatment with hydrocortisone alone.

[0045] FIG. 25 depicts the therapeutic effect of topical treatment with PINK inhibitors and steroids on mice with IMQ-induced mild-to-moderate psoriasis.

[0046] Topical treatment with DD-I and 0.25% hydrocortisone, and with DD-V and 0.25% hydrocortisone had a significantly increased therapeutic effect (p<0.01) on psoriasis, as compared to the treatment with hydrocortisone alone.

[0047] FIG. 26 depicts the synergistic and / or additive therapeutic effects that exemplary PINK inhibitors DD-L, DD-I, and DD-V have with 0.05% hydrocortisone against mild-to-moderate atopic dermatitis / eczema. Topical treatment with DD-L and 0.05% hydrocortisone, DD-I and 0.05% hydrocortisone, and with DD-V and 0.1% hydrocortisone had a significantly increased therapeutic effect (p<0.01) on atopic dermatitis / eczema, as compared to the treatment with hydrocortisone alone.

[0048] FIG. 27 depicts the synergistic and / or additive therapeutic effects that exemplary PINK inhibitors DD-L, DD-I, and DD-V have with 0.1% hydrocortisone against mild-to-moderate atopic dermatitis / eczema. Topical treatment with DD-L and 0.1% hydrocortisone, DD-I and 0.1% hydrocortisone, and with DD-V and 0.1% hydrocortisone had a significantly increased therapeutic effect (p<0.01) on atopic dermatitis / eczema, as compared to the treatment with hydrocortisone alone.

[0049] FIG. 28 depicts the synergistic and / or additive therapeutic effects that exemplary PINK inhibitors DD-I and DD-V have with 0.25% hydrocortisone against mild-to-moderate atopic dermatitis (AD)Zeczema. Topical treatment with DD-I and 0.25% hydrocortisone and with DD-V and 0.25% hydrocortisone had a significantly increased therapeutic effect (p<0.01) on atopic dermatitis / eczema, as compared to the treatment with hydrocortisone alone.

[0050] DETAILED DESCRIPTION All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

[0051] When an amount, concentration, or other value or parameter is given as either a range, preferred range or a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. Where a range of numerical values is recited herein, unless otherwise stated, the range is intended to include the endpoints thereof, and all integers and fractions within the range. It is not intended that the scope of the disclosure be limited to the specific values recited when defining a range.

[0052] As used herein, the term "about" refers to a value that is ± 5% of the stated value. In addition, it is understood that reference to a range of a first value to a second value includes the range of the stated values, e.g., a range of about 1 to about 5 also includes the more precise range of 1 to 5. It is also understood that the ranges disclosed herein include any selected subrange within the stated range, e.g., a subrange of about 50 to about 60 is contemplated in a disclosed range of about 1 to about 100.

[0053] One aspect of the present disclosure is a compound of Formula I:

[0054]

[0055] (Formula I) wherein:

[0056] R1is selected from any naturally occurring amino acid side group, an alkyl group, a cycloalkyl group, a spirocyclic group, an aryl group, an haloalkyl group, a heteroaryl group, an alkyloxy group, ora thioalkyl group; and

[0057] R2is selected from -H or an alkyl group; or a pharmaceutically acceptable salt or stereoisomer thereof.

[0058] As used herein, “a naturally occurring amino acid side group” or “a naturally occurring amino acid side chain” is any unique chemical structures attached to the alpha-carbon of naturally occurring amino acids. Each naturally occurring amino acid has a central carbon atom (the alpha carbon) to which four groups are attached: a hydrogen atom, an amino group (-NH2), a carboxyl group (-COOH), and a side chain or side group. Accordingly, R1can be selected from, but not limited to, any one of the following groups:

[0059]

[0060]

[0061] As used herein, the term “stereoisomer(s)” refers to compounds that have the same atomic connectivity but different atomic arrangement in space.

[0062] Stereoisomers can include, but are not limited to, cis-trans isomers, E and Z isomers, enantiomers, diastereomers and atropisomers. In the context of the present invention, the phrase "stereoisomer(s) thereof" is understood to mean any compound having the same connectivity as the disclosed compound but with different atomic arrangements, for example, around a chiral center. The compounds disclosed herein can be enantiomerically pure. As used herein, the term “enantiomerically pure” refers to a stereoisomer or composition of stereoisomers that possess an absolute configuration of a chiral center in an enantiomeric excess of more than 95%.

[0063] The present disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, atropisomers, tautomers and racemic and other mixtures thereof, such as enantiomers or diastereomeric enriched mixtures, all of which are within the scope of the present disclosure. Insofar as when compounds of Formula I as defined herein exist in optically active or racemic forms by virtue of one or more asymmetric carbon atoms, the phrase “stereoisomers thereof” includes in its definition any such optically active or racemic form. The synthesis of optically active compounds can be carried out by standard techniques of organic chemistry well known in the art such as, for example, by synthesis from optically active starting materials or by resolution of a racemic compound. Similarly, the enantiomeric or diastereomeric purity of a compound can be evaluated using standard laboratory techniques.

[0064] In exemplary embodiments, R1is selected from one of the following groups: and

[0065]

[0066] In exemplary embodiments, R1is selected from one of the following groups:

[0067]

[0068] As used herein, an “alkyl group” refers to any straight chain or branched, non-cyclic or cyclic, unsaturated or saturated aliphatic hydrocarbon containing from 1 to 10 carbon atoms, while the term "lower alkyl" has the same meaning as alkyl but contains from 1 to 3 carbon atoms. The term "higher alkyl" has the same meaning as alkyl but contains from 4 to 10 carbon atoms. Representative saturated straight chain alkyls include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and the like, while saturated branched alkyls include, but are not limited to, isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and the like. As used herein, a methyl substituent may be depicted as “CHs” or “Me” or as a terminal bond with no indication of specific atoms. As used herein, an “cycloalkyl group” refers to saturated and unsaturated cyclic alkyls. Representative saturated cyclic alkyls include, but are not limited to, C3-C14 (such as C3-C7) cycloalkyls, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclododecyl, and the like; while unsaturated cyclic alkyls include, but are not limited to, cyclobutenyl, cyclopentenyl and cyclohexenyl, cyclohexadiene, and the like. Cyclic alkyls are also referred to herein as "homocycles" or"homocyclic rings".

[0069] As used herein, a “spiro” or “spirocyclic group” refers to chemical structures having at least two rings sharing one common atom. The rings may be cycloalkyl, heterocyclyl or a combination thereof, and may include one or more aromatic or heteroaromatic rings. Exemplary embodiments include 1 ,4-dioxaspiro[4.5]decane, spirocyclic azetidines and spirocyclic pyrrolidines and spirocyclic piperidines, where the other ring is cycloalkyl (e.g., cyclobutane, cyclopentane or cyclohexane) or heterocyclyl (e.g., piperidine, tetrahydropyran, tetrahydrofuran, azetidine or pyrrolidine).

[0070] As used herein, an “aromatic group” or “aryl group” as used herein, refers to any aromatic carbocyclic ( / .e., all of the ring atoms are carbon) substituent such as, but not limited to, arylalkyls, aralkyls, phenyl (from benzene), tolyl (from toluene), xylyl (from xylene) or naphthyl (from naphthalene). The term "arylalkyl" or “aralkyl” as used herein refers to any alkyl having at least one alkyl hydrogen atom replaced with an aryl moiety such as, but not limited to, benzyl, -(Cl^phenyl, and -(CH2)3phenyl, -CH(phenyl)2.

[0071] As used herein, a "haloalkyl group" refers to any alkyl where at least one hydrogen atom (and including all hydrogen atoms) has been replaced with a halogen atom, such as, but not limited to, trifluoromethyl, dichloromethyl, difluoromethyl, monofluoromethyl, monobromomethyl, and 1,1,1 -trifluoroethyl.

[0072] As used herein, a "heteroaromatic group" or "a heteroaryl group" refers to any aromatic heterocycle ring of 5 to 10 or more members and having at least one heteroatom selected from nitrogen, oxygen and sulfur, and containing at least 1 carbon atom, including, but not limited to, both mono- and bicyclic- ring systems. The heteroaryl ring can be attached as a substituent via a ring heteroatom or a carbon atom. Representative heteroaromatics include, but are not limited to, furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1 ,2,4-triazole, 1 ,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1 ,2,3-oxadiazole, 1 ,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1 ,2,4-triazine, 1 ,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, and pteridine.

[0073] As used herein, an “alkyloxy group” or “alkoxy group” refers to any alkyl moiety attached through an oxygen bridge (i.e., -O-alkyl) such as, but not limited to, methoxy, ethoxy, and butoxy.

[0074] As used herein, a “thioalkyl group” means any alkyl moiety attached through a sulfur bridge (i.e., -S-alkyl) such as, but not limited to, methylthio and ethylthio.

[0075] In exemplary embodiments, R2is selected from -H and -CH2CH3.

[0076] In exemplary embodiments, R2is -CH2CH3.

[0077] As used herein, the term "pharmaceutically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal ora human.

[0078] As used herein, "pharmaceutically acceptable salt" refers to a salt that does not adversely impact the biological activity and properties of the compound and is suitable for use in contact with the tissues of subjects without undue toxicity, irritation and / or allergic response and the like. Pharmaceutically acceptable salts include those derived from suitable inorganic acids, organic acids and bases, and include, but are not limited to, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, ascorbic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, benzoic acid, naphthalene sulfonic acid, lactic acid, succinic acid, oxalic acid, and stearic acid. In some instances, pharmaceutically acceptable salts are obtained by reacting a compound having acidic group described herein with a base to form a salt such as, without limitation, an ammonium salt, an alkali metal salt (e.g., a sodium ora potassium salt), an alkaline earth metal salt (e.g., a calcium ora magnesium salt), a salt formed from an organic base, and an amino acid salt. Pharmaceutically acceptable salts derived from appropriate bases include, but are not limited to, alkali metals, alkaline earth metals, and ammonium and quaternary ammonium compounds. Specific metals include, but are not limited to, sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts can be prepared include, for example, primary, secondary, and tertiary amines.

[0079] The compounds disclosed herein have surprisingly been discovered to be capable of inhibiting the phosphorylation capabilities of a wide range of tyrosine and serine / threonine kinases involved in pro-inflammatory cellular pathways. This property of the disclosed compounds has allowed them to serve as effective agents in the treatment of diseases that arise from over-stimulation and dysregulation of the immune system (e.g., cancer, psoriasis and eczema). The disclosed compounds have also been observed to possess improved solubility, permeability, and bioavailability when compared to related compound CX-4945 (see below).

[0080] ,N

[0081]

[0082] The improvements seen with the disclosed compounds, without being bound to any particular theory, are expected to stem from the incorporation of the amide side chains. Indeed, the addition of natural amino acids at the carboxylic acid of CX-4945 has been shown to create compounds with improved PK properties. Another advantage of the claimed compounds are generally regarded as safe (e.g., when amino acids are incorporated), readily commercially available, and possess a large structure diversity. Another aspect of the present disclosure is a topical formulation including a therapeutically effective amount of a compound of Formula I; and one or more cosmetic or pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, co-surfactants, microemulsions, hydrophobic vehicles, water soluble vehicles, emulsifiers, nanoemulsifiers, buffers, humectants, moisturizers, solubilizers, preservatives, colorants, plasticizers, carriers, nanocarriers like nanocrystals, nanoparticles, ethosome, dendrimers, and liposomes, and / or excipients.

[0083] Diluents that can be included in the formulation include, but are not limited to, alcohols, glycols, oils, and water.

[0084] As used herein, the phrase "therapeutically effective amount" means any amount of a compound or pharmaceutically acceptable salt thereof of Formula I that when administered to an individual for treating a state, disease, disorder or condition is sufficient to affect such treatment, i.e., at least lessen the symptoms of said state, disease, disorder or condition. The therapeutically effective amount can vary depending on the particular state, disease, disorder or condition being treated and its severity and the age, weight, physical condition and responsiveness of the individual to be treated. Thus, one or more of these parameters can be used to select and adjust the effective amount of trolamine salicylate. Also, the amount can be determined using pharmacologic methods known in the art such as dose response curves.

[0085] In exemplary embodiments, the topical formulation contains about 1.00 wt%, 5.00 wt%, 10.00 wt%, 15.00 wt%, 20.00 wt%, 25.00 wt%, 30.00 wt%, 35.00 wt%, 40.00 wt%, 45.00 wt%, 50 wt% or any wt% falling within about 1.00 wt% to about 50.00 wt%, based on the total weight of the formulation, of a compound or pharmaceutically acceptable salt thereof of Formula I.

[0086] The topical formulation can contain one or more additives such as, but not limited to, purified water, isopropyl myristate, caprylic / capric / myristic / stearic triglyceride, dimethicone, cyclomethicone, cetyl alcohol, dimethyl sulfone, glucosamine sulfate, stearyl alcohol, glyceryl monosterate, PEG-12 glyceryl dimyristate, PEG-23 glyceryl disterate, PEG-100 stearate, glyceryl stearate, tocopheryl acetate, rosemary leaf extract, chamomilla recutita (matricaria) flower extract, lavender extract, grape seed extract, a fragrant compound or fragrant mixture, a preservative, a rheology modifier, white petrolatum, shea butter, Polysorbate 80, potassium sorbate, and / or phenoxyethanol.

[0087] In exemplary embodiments, the topical formulation contains any wt% falling within the range of about 45.00 wt% to about 95.00 wt% of purified water. In exemplary embodiments, the topical formulation contains about 45.00 wt% to about 81.00 wt% of purified water.

[0088] In exemplary embodiments, the topical formulation contains at least one component capable of producing a self-forming, thermodynamically stable liposome. Components that are capable of producing a self-forming, thermodynamically stable liposome include, but are not limited to, PEG-12 GDO, PEG-12 GDM, PEG-23 GDP, PEG-12 GDS, PEG-23 GDS, and PEG-23 glyceryl palmitate. GDO means glycerol dioleate, GDM means glycerol dimyristate, GDP means glycerol dipalmitate, and GDS means glycerol distearate.

[0089] In exemplary embodiments, the topical formulation contains one or more ionic liquids. The term “ionic liquid” has many definitions in the art, but is used herein to refer to salts (i.e., compositions including cations and anions) that are liquid at a temperature of at or below about 150° C. That is, at one or more temperature ranges or points at or below about 150° C. The ionic liquid compositions can be liquid; although, it is understood that they can be solids at other temperature ranges or points. The ionic liquids can contain at least two different ions; each of which can independently and simultaneously introduce a specific characteristic to the composition not easily obtainable with traditional dissolution and formulation techniques. Thus, by providing different ions and ion combinations, one can change the characteristics or properties of the disclosed ionic liquid compositions in a way not seen by simply preparing various crystalline salt forms. Examples of characteristics that can be controlled include, but are not limited to, melting, solubility control, and rate of dissolution. It is this multi-nature / functionality of ionic liquid compositions which allows one to fine-tune or design in very specific desired material properties. It is further understood that ionic liquid compositions can include solvent molecules {e.g., water); however, these solvent molecules should not be present in excess in the sense that the ionic liquid compositions are dissolved in the solvent, forming a solution. That is, the ionic liquid compositions contain no or minimal amounts of solvent molecules that are free and not bound or associated with the ions present in the ionic liquid composition. Thus, the ionic liquid compositions can be liquid hydrates or solvates, but not solutions. Suitable examples of cations and anions that can be included in the ionic liquid compositions include, but are not limited to, those disclosed in US 8,802,596 ( .g., pyridinium cations, pyridazinium cations, pyrimidinium cations, pyrazinium cations, imidazolium cations, pyrazolium cations, oxazolium cations and anions selected from, but not limited to, halides (e.g., fluoride, chloride, bromide, and iodide), sulfates (SC>4“), carbonates, bicarbonates, phosphates, phosphates, nitrates (NOs-), nitrites (NO2"), acetates (CH3CO2"), PFe" , BFr , triflate (TfO; CF3SO2’), nonaflate (NfO; CF3(CF2)3SO2’), bis(triflyl)amide (Tf2N; (CFsSC^N-), trifluoroacetate (TA; CFsCCh”), and heptafluororobutanoate (HB; CF3(CF2)3SO2"), the content of which is incorporated herein in its entirety by reference.

[0090] In exemplary embodiments, the topical formulation contains at least one extracellular matrix component, fragments thereof and / or combinations thereof. Extracellular matrix (ECM) components can be divided into several classes of biomolecules based upon their structure and function within an extracellular matrix. The most prominent class is the structural class of extracellular matrix proteins. These include the collagen and elastin families of proteins. Collagen fibers can strengthen and organize the matrix; elastin fibers can provide flexibility and resilience. Another class is of specialized proteins, such as fibrillin, fibronectin, laminin, merosin, tenascin, and vitronectin, which serve less of a structural role and more of an adhesive or integral role within the extracellular matrix. These proteins allow for cell attachment and form crosslinks within the matrix gel. Numerous proteoglycans and heparan sulfate containing proteins can form the highly hydrated gel-like mixture that helps stabilize the matrix within an aqueous environment. Proteoglycans include a protein core to which are attached long chains of glycosaminoglycans (GAGs) forming extremely complex high molecular weight components of the ECM. Another GAG which can be a component of an extracellular matrix is hyaluronic acid, a non-sulfate GAG. The extracellular matrix component can be, but is not limited to, proteins, peptides, or a receptor associated with an extracellular matrix, hyaluronic acid, elastin, collagen, fibronectin, lectin, and fragments thereof and combinations thereof. Such fragments can include, without limitation, hyaluronic acid fragments, collagen fragments, fibronectin fragments, elastin fragments, lectin fragments, and combinations thereof.

[0091] In exemplary embodiments, the topical formulation contains from about 0.001 wt% to about 10 wt%, from about 0.1 wt% to about 2.0 wt%, about 0.25 wt% to about 3.0 wt%, about 0.5 wt% to about 5.0 wt%, about 0.75 wt% to about 7.5 wt%, based on the total weight of the formulation, of one or more extracellular matrix components.

[0092] In exemplary embodiments, the topical formulation contains one or more enzymes capable of facilitating delivery of a compound or pharmaceutically acceptable salt thereof of Formula I through one or more layers of skin. These enzymes can include, but are not limited to, a hyaluronidase and an elastase.

[0093] These enzymes can be present in the topical formulation in concentrations ranging from about 0.1 wt% to about 25 wt%, about 0.1 wt% to about 20 wt%, about 0.1 wt% to about 15 wt%, about 0.1 wt% to about 10 wt%, about 0.1 wt% to about 8 wt%, about 0.1 wt% to about 5 wt%, about 0.1 wt% to about 4 wt%, about 0.1 wt% to about 3 wt%, about 0.1 wt% to about 3 wt%, or about 0.1 wt% to about 1 wt% based on the total weight of the formulation. In other exemplary embodiments, the enzymes are present in concentrations of about 0.1 wt%, about 0.5 wt%, about 1 wt%, about 2 wt%, about 5 wt%, about 10 wt%, or about 25 wt%, based on the total weight of the formulation.

[0094] In exemplary embodiments, the topical formulation contains one or more additional active agents. Non-limiting examples of active agents include a biologic, therapeutic peptides, biomimetic peptide, small molecule and macromolecular analgesic agents, antifungal agents, antibacterial agents, anesthetic agents, proteins, prostaglandins, enzyme inhibitors, steroids, small molecule drugs, macromolecular drugs, biologies, antibodies, chimeric antibodies, antibody fragments, diagnostic antibodies, antigens, peptides, adjuvants, antioxidants, cosmetic ingredients, therapeutic cells, diagnostic agents, radioactive tracers, contrast agents, neurotoxins, sensation modifying agents, and combinations thereof.

[0095] In exemplary embodiments, the topical formulation contains one or more sensation modifying agents. Examples of sensation modifying agents include, but are not limited to, a cooling agent, a warming agent, a relaxing or soothing agent, a stimulating or refreshing agent, and mixtures thereof.

[0096] The cooling agent can be selected from, but is not limited to, menthol; an isomer of menthol, a menthol derivative; 4-Methyl-3-(1-pyrrolidinyl)-2[5H]-furanone; WS-23, Icilin, Icilin Unilever Analog, 5-methyl-4-(1-pyrrolidinyl)-3-[2H]-furanone; 4,5-dimethyl-3-(1-pyrrolidinyl)-2[5H]-furanone; isopulegol, 3-(1-menthoxy)propane-1,2-diol, 3-(1-menthoxy)-2-methylpropane-1 ,2-diol, p-menthane-2,3-diol, p-menthane-3,8-diol, 6-isopropyl-9-methyl-1,4-dioxas-piro[4,5]decane-2-methanol, menthyl succinate and its alkaline earth metal salts, trimethylcyclohexanol, N-ethyl-2-isopropyl-5-methylcyclohexanecarb-oxamide, Japanese mint (Mentha arvensis) oil, peppermint oil, menthone, menthone glycerol ketal, menthyl lactate, 3-(1-menthoxy)ethan-1-ol, 3-(1-menthoxy)propan-1-ol, 3-(1-menthoxy)butan-1-ol, 1-menthylacetic acid N-ethyl amide, 1-menthyl-4-hydroxypentanoate, 1-menthyl-3-hydroxybutyrate, N, 2, 3-trimethyl-2-(1 -methyl ethyl)-butanamide, spearmint oil and combinations thereof.

[0097] The warming agent can be selected from, but is not limited to, polyhydric alcohols, capsaicin, capsicum powder, a capsicum tincture, capsicum extract, capsaicin, hamamalis, homocapsaicin, homodihydrocapsaicin, nonanoyl vanillyl amide, nonanoicacid vanillyl ether, vanillyl alcohol alkyl ether derivatives, such as vanillyl ethyl ether, vanillyl butyl ether, vanillyl pentyl ether, and vanillyl hexyl ether, isovanillyl alcohol alkyl ethers, ethylvanillyl alcohol alkyl ethers, veratryl alcohol derivatives, substituted benzyl alcohol derivatives, substituted benzyl alcohol alkyl ethers, vanillin propylene glycol acetal, ethylvanillin propylene glycol acetal, ginger extract, ginger oil, gingeol, gingeron and combinations thereof.

[0098] The relaxing or soothing agent can be selected from, but is not limited to, herb extracts, selected from the group including of aloe vera, alpha bisabolol, D-panthenol, allantoin, hamamelis, chamomile, yarrow; calendula, comfrey, witch hazel and other astringents, sea weed, and oat extracts; oils, selected from the group consisting of: almond oil, avocado oil, and comfrey; and essential oils, selected from the group consisting of: cardamone, eucalyptus, mentha piperita (peppermint), hyssop, and rosemary; waxy or unctuous substances selected from the group consisting of: lanolin or vaselline jelly, minerals, selected from the group consisting of: zinc oxide, calamine and selenium; vitamins, selected from the group consisting of: tocopheryl acetate (vitamin E), and pharmaceutical agents selected from the group consisting of: analgesics, anesthetics, antiinflammatory agents, and anti-histamines, and muscle relaxants; menthol, camphor, eugenol, eucalyptol, safrol, methyl salicylate, menthyl lactate, menthyl ethoxyacetate, menthone glycerinacetal, 3-1-menthoxypropane-1,2-diol, ethyl 1-menthyl carbonate, (1S,3S,4R)-p-menth-8-en-3-ol, menthyl pyrrolidone carboxylate, N-substituted-p-menthane-3-carboxamides hamamelis extract, ginger oil and combinations thereof.

[0099] The topical formulations disclosed herein can be formulated as paste, gel, ointment, solutions, lotion, emulsion, microemulsion, cream, foam, mousse, spray, aerosols, suspension, dispersion, tapes, ionic liquid ora liposomal formulation.

[0100] In exemplary embodiments, the topical formulation contains one or more cosmetic or pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic vehicles, water soluble vehicles, nanosuspension, emulsifiers, nano emulsifiers, buffers, humectants, moisturizers, solubilizers, preservatives, colorants, plasticizers, carriers, nanocarriers, dendrimers, liposomes, excipients, and combinations thereof. The person of ordinary skill in the art can refer to various pharmacologic references such as, for example, Modern Pharmaceutics, Banker & Rhodes, Marcel Dekker, Inc. (1979) and Goodman & Gilman's The Pharmaceutical Basis of Therapeutics, 6th Edition, MacMillan Publishing Co, New York (1980) for guidance in determining the amount of such components in the formulation.

[0101] Excipients that can be included in the formulation can be selected from, but are not limited to, gelatin, casein, lecithin, gum acacia, cholesterol, tragacanth, stearic acid, benzalkonium chloride, calcium stearate, glyceryl monostearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan esters, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyethylene glycols, polyoxyethylene stearates, colloidol silicon dioxide, phosphates, sodium dodecyl sulfate, carboxymethylcellulose calcium, carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethycellulose phthalate, noncrystalline cellulose, magnesium aluminum silicate, triethanolamine, polyvinyl alcohol, polyvinylpyrrolidone, sugars, starches and combinations thereof.

[0102] Surfactants (i.e., surface-active agents that lower surface tension and thereby increase the emulsifying, foaming, dispersing, spreading and wetting properties of a product) that can be include in the formulation can be selected from, but are not limited to, emulsifying wax, glyceryl monooleate, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polysorbate, sorbitan esters, benzyl alcohol, benzyl benzoate, cyclodextrins, glycerin monostearate, poloxamer, povidone, stearyl alcohol and combinations thereof.

[0103] Emulsifiers (i.e., surface active substances which promote the suspension of one liquid in another and / or promote the formation of a stable mixture, emulsion, nanoemulsion, or microemulsion, of oil and water) that can be included in the formulation can be selected from, but are not limited to, metallic soaps, animal and vegetable oils, acacia, anionic emulsifying wax, calcium stearate, carbomers, cetostearyl alcohol, cetyl alcohol, cholesterol, diethanolamine, ethylene glycol palm itostea rate, glycerin monostearate, glyceryl monooleate, hydroxpropyl cellulose, hypromellose, lanolin, hydrous, lanolin alcohols, lecithin, medium-chain triglycerides, methylcellulose, mineral oil and lanolin alcohols, monobasicsodium phosphate, monoethanolamine, nonionic emulsifying wax, oleic acid, poloxamer, poloxamers, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, propylene glycol alginate, self-emulsifying glyceryl monostearate, sodium citrate dehydrate, sodium lauryl sulfate, sorbitan esters, stearic acid, sunflower oil, tragacanth, triethanolamine, xanthan gum, glycerol stearate and combinations thereof.

[0104] In exemplary embodiments, the topical formulation is formulated as oil-in-water or water-in-oil emulsion or microemulsion, cream or ointment. A cream can be a water-in-oil (w / o) emulsion or microemulsion in which an aqueous phase is dispersed in an oil phase, or an oil-in-water (o / w) emulsion or microemulsion in which an oil is dispersed within an aqueous base. Creams of the topical pharmaceutical formulation can contain emulsifying agents and / or other stabilizing agents. In exemplary embodiments, the topical formulation is a cream having a viscosity of greater than 1000 centistokes, or a viscosity in the range of about 20,000-50,000 centistokes. An ointment generally refers to a more viscous oil-in-water cream. Ointment bases ( / .e. carriers) that can be used in the formulation include, but are not limited to, hydrocarbons (petrolatum, beeswax), vegetable oils, fatty alcohols (cholesterol, lanolin, wool alcohol, stearyl alcohol) or silicones.

[0105] Insoluble solids such as starch, zinc oxide, calcium carbonate, or talc can also be used in embodiments wherein the formulation is formulated as an ointment or cream.

[0106] In exemplary embodiments, the topical formulation is formulated as a lotion. A “lotion”, as used herein, is a low- to medium-viscosity liquid composition. A lotion can contain finely powdered substances that are soluble in the dispersion medium through the use of suspending agents and dispersing agents. Alternatively, lotions can have, as the dispersed phase, liquid substances that are immiscible with a vehicle and can dispersed by means of emulsifying agents or other suitable stabilizers. In exemplary embodiments, the topical formulation is a lotion in the form of an emulsion or microemulsion having a viscosity of between 100 and 1000 centistokes. The fluidity of lotions permits rapid and uniform application over a wide surface area.

[0107] In exemplary embodiments, the topical formulation is formulated as a gel. Gel forms of the topical formulation can include a lipophilic component, an aqueous component or both. In exemplary embodiments, the topical formulation includes one or more gelling agents selected from, but not limited to, modified celluloses, such as hydroxypropyl cellulose and hydroxyethyl cellulose; carbopol homopolymers and copolymers; and combinations thereof. In exemplary embodiments, the topical formulation is formulated as a gel and includes one or more liquid vehicles selected from, but not limited to, diglycol monoethyl ether; alklene glycols, such as propylene glycol; dimethyl isosorbide; alcohols, such as isopropyl alcohol, ethanol and combinations thereof. The solvents can be selected for their ability to dissolve the other components included in the gel formulation. Other additives can be included in the gel dosage forms of the formulations, such as, but not limited to, isopropyl myristate, ethyl acetate, C12-C15 alkyl benzoates, mineral oil, squalane, cyclomethicone, capric / caprylic triglycerides, or combinations thereof.

[0108] Gel forms of the formulations can be formed by the entrapment of large amounts of aqueous or aqueous-alcoholic liquids in a network of polymers or of colloidal solid particles. Such polymers or colloids (gelling or thickening agents) can be present at concentrations of less than 10 wt%, based on the total weight of the formulation, and include, but are not limited to, carboxymethyl cellulose, hydroxypropylmethyl cellulose, hydroxyethyl cellulose, methyl cellulose, sodium alginate, alginicacid, pectin, tragacanth, carrageen, agar, clays, aluminum silicate, carbomers, and combinations thereof.

[0109] In exemplary embodiments, the topical formulation includes one or more vitamins, cosmetic peptides, oil control agents, and / or other skin care agents.

[0110] Vitamins that can be included in the topical formulation include, but are not limited to, vitamin D, vitamin K, vitamin B (including niacinamide, nicotinic acid, Ci-18 nicotinic acid esters, and nicotinyl alcohol; B6 compounds, such as pyroxidine; and B5 compounds, such as panthenol, or“pro-B5”), vitamin A (including retinoids such as retinyl propionate, carotenoids, and other compounds), vitamin E (including tocopherol sorbate, tocopherol acetate, other esters of tocopherol), vitamin C (including ascorbyl esters of fatty acids, and ascorbic acid derivatives, for example, ascorbyl glucoside, magnesium ascorbyl phosphate, sodium ascorbyl phosphate, and ascorbyl sorbate), and all natural and / or synthetic analogs thereof, and combinations thereof. In exemplary embodiments, the formulation includes about 0.0001 wt% to about 50 wt%, about 0.001 wt% to about 10 wt%, about 0.01 wt% to about 5 wt%, or about 0.1 wt% to about 1 wt%, based on the total weight of the formulation, of one or more vitamins.

[0111] Cosmetic peptides that can be included in the topical formulation include, but are not limited to, di-, tri-, tetra-, penta-, and hexa-peptides, their salts, isomers, derivatives, and mixtures thereof. Examples of useful peptide derivatives include, but are not limited to, peptides derived from soy proteins, palmitoyl-lysine-threonine (pal-KT) and palmitoyl-lysine-threonine-threonine-lysine-serine (MATRIXYL®) palmitoyl-glycine-glutamine-proline-arginine (RIGIN®), these three being available from Sederma, France, and Cu-histidine-glycine-glycine (Cu-HGG, also known as IAMIN®), and naturally occurring and synthesized derivatives thereof, and combinations thereof. In exemplary embodiments, the formulation includes about 1xio-7wt% to about 20 wt%, about 1x10“6wt% to about 10 wt%, about 1xio-5wt% to about 5 wt%, based on the total weight of the formulation, of one or more cosmetic peptides.

[0112] Oil control agents that can be included in the topical formulation include, but are not limited to, salicylic acid, dehydroacetic acid, benzoyl peroxide, vitamin B3 (for example, niacinamide), their isomers, esters, salts and derivatives, and mixtures thereof. In exemplary embodiments, the formulation includes about 0.0001 wt% to about 15 wt%, about 0.01 wt% to about 10 wt%, about 0.1 wt% to about 5 wt%, about 0.2 wt% to about 2 wt%, based on the total weight of the formulation, of one or more oil control agents.

[0113] Other skin care agents that can be included in the topical formulation include, but are not limited to, retinol, steroids, sunblock, salicylate, minocycline, antifungals, peptides, antibodies, lidocaine, N-acyl amino acid compounds (including, for example, N-acyl phenylalanine, N-acyl tyrosine, and their isomers, including their D and L isomers) along with their salts and derivatives, and combinations thereof. An exemplary N-acyl amino acid compound is N-undecylenoyl-L-phenylalanine (commercially available under the tradename SEPIWHITE®). Other skin active agents such as Lavandox, Thallasine 2, Argireline NP, Gatuline In-Tense and Gatuline Expression, Myoxinol LS 9736, Syn-ake, and Instensyl®, Sesaflash™, N-acetyl D-glucosamine, panthenol (for example, DL panthenol available from Alps Pharmaceutical Inc.), tocopheryl nicotinate, benzoyl peroxide, 3-hydroxy benzoic acid, flavonoids (for example, flavanone, chaicone), farnesol, phytantriol, glycolic acid, lactic acid, 4-hydroxy benzoic acid, acetyl salicylic acid, 2-hydroxybutanoic acid, 2-hydroxypentanoic acid, 2-hydroxyhexanoic acid, cis-retinoic acid, trans-retinoic acid, retinol, retinyl esters (for example, retinyl propionate), phytic acid, N-acetyl-L-cysteine, lipoic acid, tocopherol and its esters (for example, tocopheryl acetate: DL-a-tocopheryl acetate available from Eisai), azelaic acid, arachidonic acid, tetracycline, ibuprofen, naproxen, ketoprofen, hydrocortisone, acetominophen, resorcinol, phenoxyethanol, phenoxypropanol, phenoxyisopropanol, 2,4,4'-trichloro-2'-hydroxy diphenyl ether, 3,4,4'-trichlorocarbanilide, octopirox, lidocaine hydrochloride, clotrimazole, miconazole, ketoconazole, neomycin sulfate, theophylline, and mixtures thereof can also be included within the formulation. Additional skin care agents that can be included in the formulation are disclosed in US Publication No. 2007 / 0020220A1 , wherein the components / ingredients are incorporated herein by reference in their entirety.

[0114] In exemplary embodiments, the topical formulation includes one or more skin lightening agents, such as, but not limited to, ascorbic acid compounds, vitamin B3 compounds, azelaic acid, butyl hydroxyanisole, gallic acid and its derivatives, glycyrrhizinic acid, hydroquinone, kojic acid, arbutin, mulberry extract, and mixtures thereof.

[0115] In exemplary embodiments, the topical formulation includes one or more sunblock agents such as, but not limited to, para-aminobenzoic acid (PABA), PABA esters (glyceryl PABA, amyldimethyl PABA and octyldimethyl PABA), benzophenones (oxybenzone and sulisobenzone), cinnamates (octylmethoxy cinnamate and cinoxate), salicylates (homomethyl salicylate) anthranilates, TiOs, avobenzone, bemotrizinol, bisoctrizole, 3-(4-methylbenzylidene)-camphor, cinoxate, diethylamino hydroxybenzoyl hexyl benzoate, dioxybenzone, drometrizole trisiloxane, ecamsule, ethylhexyl triazone, homosalate, menthyl anthranilate, octocrylene, octyl salicylate, iscotrizinol, isopentenyl-4-methoxycinnamate, octyl-dimethyl-p-aminobenzoic acid, octyl-methoxycinnamate, oxybenzone, polysilicone-15, trolamine salicylate, ZnO and combinations thereof.

[0116] In exemplary embodiments, the topical formulation is a hydrogel. Hydrogel dosage forms of the topical formulation can be prepared by cross-linking various monomers and / or polymers to provide a three-dimensional polymer network. Nonlimiting examples of cross-linkable polymers include, but are not limited to, polyoxyethylene-polypropylene block copolymers, ionic poly saccharides, such as chitosan or sodium alginate, cellulose, and biodegradable polymers, such as polylactides (PLA) and poly-glycolides (PGA), butylene succinate (PBS), polyhydroxyalkanoate (PHA), polycaprolactone acid lactone (PCL), polyhydroxybutyrate (PHB), glycolic amyl (PHV), PHB and PHV copolymer (PHBV), and poly lactic acid (PLA)-polyethylene glycol (PEG) copolymers (PLEG).

[0117] In exemplary embodiments, the topical formulation contains one or more penetration or permeation enhancers for transdermal drug delivery. Non-limiting examples of penetration or permeation enhancers include chemical permeation enhancers (CPEs) such as, but not limited to, solvents (e.g., monohydric alcohols such as methanol, ethanol, propanol, isopropanol), fatty acids (e.g., oleic acid, caprylic acid), fatty alcohols (e.g. lauryl alcohol, myristyl alcohol, oleyl alcohol), surfactants (e.g., ionic or non-ionic detergents), fatty acid esters e.g. isopropyl myristate, isopropyl palmitate, methylpropionate, and ethyl oleate), organic acids (e.g. salicylic acid and salicylates, citric acid and succinic acid), nitrogenous compounds (e.g. urea), bile salts and derivatives, micelles / liposomes or micelleforming or liposome-forming components (e.g., phospholipids), sulfoxides, terpenes and terpenoids, polyols, urea and derivatives, and chelating agents. A non-limiting list of CPEs can also include methanol, ethanol, propylene glycol, ethylene glycol, glycerol, butanediol, polyethylene glycol, polyethylene glycol monolaurate, diethylene glycol, monoethyl ether (transcutol), oleic acid, caprylic acid, menthol, nerol, camphor, methyl salicylate, sodium laurate, sodium lauryl sulfate, cetyltrimethylammonium bromide, benzalkonium chloride, poloxamer (231, 182, 184), Tween (20, 40, 60, 80), sodium dodecyl sulfonate (SDS), methylsulfonylmethane (MSM), benzalkonium chloride, polyoxyl 40 hydrogenated castor oil, didecyldimethylammonium bromide (DDAB), didecyltrimethylammonium bromide (DTAB), polysorbates, Na glyacolate, Na deoxycholate, EDTA, citric acid, dimethylacetamide (DMA), DMSO, dimethylformamide (DMF), dimethylsulfoxide, decylmethylsulfoxide, propylene glycol, polyethylene glycol, glycerol, 2-pyrrolidone, 1-methyl-2-pyrrolidone, ethanolamine, diethanolamine, triethanolamine urea, lecithin, terpenes, terpenoids, 1 -substituted azacycloheptan-2-ones, such as 1-n-dodecylcyclazacycloheptan-2-one phospholipids, water, and combinations thereof. More permeation enhancers that can be used in the formulation are those disclosed in, for example, Williams et al., “Penetration enhancers,” Adv. Drug Deliv. Rev. 2004 Mar. 27; 56(5):603-18; and Pathan et al., “Chemical Penetration Enhancers for Transdermal Drug Delivery System,” Trap. J. Pharma. Res., April 2009; 8(2): 173-179, the contents of which are incorporated herein by reference in their entireties.

[0118] In exemplary embodiments, the topical formulation contains about 0.1 wt% to about 15 wt%, about 2 wt% to about 12 wt%, about 4 wt% to about 10 wt%, about 4 wt% to about 7 wt%, about 4 wt% to about 6 wt%, about 4.5 wt% to about 5.5 wt%, about 4 wt% to about 5 wt%, based on the total weight of the formulation, of one or more penetration or permeation enhancers.

[0119] One or more of the penetration or permeation enhancers that can be included in the formulation can be a non-ionizable glycol ether. Non-ionizable glycol ethers include, but are not limited to, diethylene glycol monomethyl ether, triethylene glycol monomethyl ether, polyethylene glycol monomethyl ether, ethylene glycol monoethyl ether, diethylene glycol monoethyl ether, triethylene glycol monoethyl ether, ethylene glycol monoisopropyl ether, ethylene glycol monobutyl ether, diethylene glycol monobutyl ether, triethylene glycol monobutyl ether, ethylene glycol monoisobutyl ether, diethylene glycol monohexyl ether, ethylene glycol mono 2-ethylhexyl ether, diethylene glycol mono 2-ethylhexyl ether, ethylene glycol monoallyl ether, ethylene glycol monophenyl ether, ethylene glycol monobenzyl ether, diethylene glycol monobenzyl ether, propylene glycol monomethyl ether, dipropylene glycol monomethyl ether, tripropylene glycol monomethyl ether, dipropylene glycol monopropyl ether, propylene glycol monobutyl ether, dipropylene glycol monobutyl ether, propylene glycol monophenyl ether, ethylene glycol dimethyl ether, diethylene glycol dimethyl ether, triethylene glycol dimethyl ether, diethylene glycol diethyl ether, diethylene glycol dibutyl ether, dipropylene glycol dimethyl ether, and combinations thereof. The glycol portion of these non-ionizable glycol ethers can include a broad range of chemicals including, but not limited to, propylene glycol, dipropylene glycol, butylene glycol, and polyethyleneglycols having a general formula of HOCH2(CH2OH)nCH2OH, where n (number of oxyethylene groups) is 4-200. In exemplary embodiments, the non-ionizable glycol ether is a diethylene glycol monoethyl ether (“DEGEE” or “ethoxydiglycol” known under its trade name TRANSCUTOL®, commercially available from Gattefosse, Paramus, N.J). In exemplary embodiments, the topical formulation contains about 0.01 wt% to 50 wt%, about 0.5 wt% to about 10 wt%, about 1 wt% to about 25 wt%, about 2 wt% to about 20 wt%, about 4 wt% to about 10 wt%, about 4 wt% to about 8 wt%, about 4 wt% to about 5 wt%, based on the total weight of the formulation, of one or more non-ionizable glycol ethers.

[0120] One or more of the penetration or permeation enhancers that can be included in the formulation can be a peptide or protein fragment. Such peptides and protein fragments can be classified as “skin penetrating peptides” (SPPs) or cell penetrating peptides (CPPs). SPPs can stabilize structural proteins in the skin rather than denaturing them. For example, SPPs bind to keratin proteins through hydrogen bonds and weak electrostatic interactions and can operate as binding mediators between keratin and drug molecules. SPPs can also utilize pathways between corneocytes via diffusion of a drug via gaps between cells as well as through lipid bilayers, without disruption. An example of a SPP is TD-1 , which is known to loosen the desmosome-induced tight junctions between corneocytes with a change in the space between cells from about 30 nm to about 466 nm in 30 minutes from topical application. The cell gaps increase and then gradually are restored in 1 hour after treatment with TD-1. Various SPPs are known in the art and numerous peptides containing 9 to 19 amino acids have been shown to exhibit skin penetrating activity. Embodiments encompass all such peptides.

[0121] In exemplary embodiments, the topical formulation contains one or more fragrance-inducing compounds. As used herein, a “fragrance-inducing compound” is a chemical that provides the topical pharmaceutical formulation a specific aroma. The fragrance-inducing compound can be selected from, but is not limited to, ethyl acetate, benzaldehyde, ethyl maltol, limonene, kashmirin, and acetone. In exemplary embodiments, the topical formulation contains one or more fragranceinducing compounds that provide the formulation an aroma or scent selected from, but not limited to, tropical, sage, peppermint, lemon, eucalyptus, bergamot, tea tree, lemongrass, grapefruit, coconut, vanilla, lavender, bay rum, cedarwood, musk, vetiver, pine, black pepper, and sandalwood. Those of ordinary skill in the art readily understand what chemicals and essential oils are capable of achieving the above exemplified aromas and scents are readily capable of blending those fragrance-inducing compounds with other components of the topical formulation.

[0122] In exemplary embodiments, the topical formulation is in the form of a transdermal patch. The transdermal patch can be in any conventional form such as, but not limited to, a strip, a gauze, or a film. The transdermal patch can contain nonwoven or woven (e.g., gauze dressing) material. The transdermal patch can contain laminated layers. The transdermal patch can be nonocclusive or occlusive, with the latter being preferred, but not required, for backing layers. The transdermal patch can be hermetically sealed for storage (e.g., foil packaging). The transdermal patch can be held onto the skin and components of the patch can be held together using various adhesives. For example, the transdermal patch can be in the form of a band-aid type device, or it may be packaged in a small metal or plastic “cup”, which is strapped onto an appropriate site using an adhesive, tape, or an outer fabric or leather strap, similar to that worn as part of a watch. The transdermal patch be disposable or can be refillable / reusable.

[0123] The transdermal patch can be made of any polymeric material, non-limiting examples including ethylene vinyl acetate (EVA) copolymer, crosslinked poly(vinyl alcohol), poly(hydroxy ethylmethacrylate), acyl substituted cellulose acetates, hydrolyzed alkylene-vinyl acetate copolymers, polyvinyl chloride, polyvinyl acetate, polyvinyl alkyl ethers, polyvinyl fluoride, polycarbonate, polyurethane, polyamide, polysulphones, styrene acrylonitrile copolymers, crosslinked polyethylene oxide), poly(alkylenes), poly(vinyl imidazole), poly(esters), poly(ethylene terephthalate), polyphosphazenes, chlorosulphonated polyolefines, poly-lactides (PLA), polyglycolides (PGA), and combinations thereof.

[0124] In exemplary embodiments, the transdermal patch contains a hydrogel. In other exemplary embodiments, the transdermal patch includes additional components such as, but not limited to, binders, buffers, colorings, desiccants, diluents, humectants, preservatives, stabilizers, other excipients, adhesives, plasticizers, tackifiers, thickeners, cooling agents, and combinations thereof.

[0125] In exemplary embodiments, the topical formulation contains one or more steroids. Steroids that can be included in the formulation include, but are not limited to, hydrocortisone, clobetasone, betamethasone, triamcinolone, fluocinolone acetonide, fluticasone propionate, mometasone furoate, hydrocortisone butyrate, hydrocortisone valerate, halobetasol, amcinonide, betamethasone dipropionate, diflucortolone valerate, beclomethasone, and other mineralocorticoids and glucocorticoids.

[0126] In exemplary embodiments, the formulation contains one or more natural anti-inflammatory agents. As used herein, a “natural anti-inflammatory agent” refers to a compound derived from a natural source (i.e., an animal or plant) that is known to produce an anti-inflammatory effect when administered to an animal or human. Examples of natural anti-inflammatory agents include, but are not limited to, curcumin, ginger, allicin, capsaicin, omega-3 fatty acids, vitamin D, green tea extract, and salicin.

[0127] Another aspect of the present disclosure is a method for treating a skin condition in a subject in need thereof, the method including administering to the subject in need of treatment for a skin condition a topical composition containing a therapeutically effective amount of a compound or pharmaceutically acceptable salt thereof of Formula I; and one or more cosmetic or pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic vehicles, water soluble vehicles, emulsifiers, buffers, humectants, moisturizers, solubilizers, preservatives, colorants, plasticizers, carriers, nanocarriers, dendrimers, liposomes, and / or excipients.

[0128] In exemplary embodiments, the topical composition is a topical formulation disclosed herein.

[0129] In exemplary embodiments, the skin condition is characterized by one or more inflammation, itching, redness, rapid skin growth, thickening of skin, blistering of the skin or aging of the skin. Possible skin conditions characterized by inflammation, itching, redness, rapid skin growth, thickening of skin, blistering of the skin and aging of the skin include, but are not limited to, mild-to-severe atopic dermatitis (eczema), mild-to-severe psoriasis, Lupus, dermatomyositis, Lichen Planus, Pemphigus, Hidradenitis suppurativa, Vitiligo, Scleroderma, Pemphigoid, wrinkles, sagging skin, thinning skin, loss of skin elasticity and firmness, and appearance of age spots and dark spots. Another aspect of the present disclosure is a method for treating a malignant disease or autoimmune disorder in a subject in need thereof, the method includes administering to a subject in need of treatment for a malignant disease or autoimmune disorder a composition containing a therapeutically effective amount of a compound or pharmaceutically acceptable salt thereof of Formula I.

[0130] In exemplary embodiments, the subject in need thereof suffers from an autoimmune disorder selected from mild-to-severe atopic dermatitis (eczema), mild-to-severe psoriasis, Lupus, dermatomyositis, Lichen Planus, Pemphigus, Hidradenitis suppurativa, Vitiligo Scleroderma, and Pemphigoid.

[0131] In exemplary embodiments, the subject in need thereof suffers from a malignant disease selected from a topical or skin cancer, for example, basocellular carcinoma, squamous cell carcinoma and melanoma and other cancers having the mode of action that follows the tyrosine kinase pathways.

[0132] In exemplary embodiments, the subject in need thereof suffers from a malignant disease selected from a blood cancer, for example, leukemia or lymphoma, or lung, breast, colon, prostate, bladder, kidney, ovarian, and pancreatic cancers, or any type of cancer driven by increased activity of tyrosine kinase(s) and / or serine / threonine kinase(s).

[0133] In exemplary embodiments, the composition is administered topically or systemically to the subject in need thereof.

[0134] In exemplary embodiments, the composition is formulated for intravenous or systemic administration (e.g., subcutaneous administration and intramuscular administration). In these embodiments, the composition can be in the form of an injectable composition or intravenous infusion (e.g., solutions, suspensions, and emulsions). For parenteral administration, the composition can include a carrier that contains sterile water and optionally other ingredients to aid in solubility. Injectable solutions can be prepared from a carrier that contains a saline solution, a glucose solution, or a mixture of a saline and a glucose solution. Compositions for intravenous or systemic administration can include a compound of Formula I, or a pharmaceutically acceptable salt or stereoisomer thereof, in an amount in a range of about 100 micrograms to about 2000 micrograms. Given the prevalence of intravenous or systemic drug administration in the art, those skilled in the art are readily capable of developing intravenous or systemic administration forms of the compositions disclosed herein.

[0135] In exemplary embodiments, the method includes administering one or more steroids, such as topical corticosteroids (e.g., hydrocortisone), with or after administration of the composition.

[0136] Examples

[0137] The present disclosure will be described in more detail with reference to the following Examples, which shows exemplary embodiments in accordance with the present disclosure. The present disclosure is not limited to these exemplary embodiments.

[0138] Example 1 : Topical PINK Inhibitors Elicit a Therapeutic Effect on IMQ-induced Psoriasis Preclinical Model via the Influence of Multiple Kinases Activities Animal Treatment and Cryosections

[0139] Female C57BL / 6J mice (7-8 weeks old, Jackson Laboratory stock #000664) were housed under specific pathogen-free conditions, received daily topical applications of 5% imiquimod (IMQ) cream (Perrigo) on both ears for 1 through 3 days (see FIG. 1). PINK inhibitors DD-V (- valine ester), DD-L (- leucine ester), and DD-I (-isoleucine ester) were administered as a Proguard™ cream formulation containing 15% w / v of the inhibitor. DMSO was administered as a control (10 pL). The structures of DD-V, DD-L, and DD-I are provided below.

[0140]

[0141]

[0142] (DD-I)

[0143] The three PINK inhibitors were prepared by the following approach. Briefly, the corresponding ethyl esters of select amino acid hydrochlorides (1.2 mmol) were neutralized with N,N-diisopropylethylamine (3 mmol) in a suspension of ethyl acetate (15 mL) as solvent for 16 hours. Next TBTU (1.4 mmol), (5-(3-chlorophenylamino)benzo [c][2,6]naphthyridine-8--carboxylic acid (1 mmol), and DMSO (2 mL) was added. The reaction mixture was then heated at a temperature of 60°C for 18 hours. The reaction mixture was cooled to room temperature and quenched with water. The organic products of the reaction were extracted with ethyl acetate (3 x 40 mL). The organic layers were combined, washed with water (3 x 25 mL), dried over MgSC , filtered, and evaporated to yield corresponding conjugated amino acid esters. Column chromatography on a silica gel column using methylene chloride: methanol (9.5:0.5) as eluent gave the quantitative yield of desired DD-V (-valine ester), DD-L (-leucine ester), and DD-I (-isoleucine ester). Characterization and structures were confirmed by NMR and MS analysis. Preliminary analytical studies showed that the novel PINK inhibitors were 150mg -175 mg / mL soluble in DMSO. For reference the parent compound 5-(3-chlorophenylamino)benzo [c][2,6]naphthyridine-8-caboxylic acid has a solubility of 16 mg / mL in DMSO (Selleckchem.com data sheet, Houston, TX, USA). In addition, the novel PINK inhibitors were more stable at room temperature, relatively easy to manufacture, and were highly compatible with large-scale purification by chromatography due to their improved solubility in organic solvents.

[0144] The PINK inhibitors and DMSO were applied twice daily, starting 45 minutes post-IMQ treatment (see FIG. 1). Kinome analysis was performed on day 1, day 2 and day 3 after the application of IMQ and beginning of the treatment with the respective PINK inhibitor (DD-L, DD-I, or DD-V) (see FIG. 1). Ears were harvested 24 hours following the final treatment, embedded in Tissue Plus ® OCT compound (Fisher Healthcare), and flash-frozen in liquid nitrogen. Cryosectioning was performed using a Leica cryostat maintained at -20°C, with three consecutive 10 pm epidermal sections collected from the dorsal treated skin surface of each ear using low-profile Accu-Edge® Disposable Microtome Blades (Sakura Finetek). Sections were stored at -80°C in pre-cooled microcentrifuge tubes until further analysis.

[0145] Kinase Activity Profiling

[0146] Kinome profiling was performed at PamGene International BV, Netherlands. PamGene’s KinomePro approach measures the ability of active kinases in a cell lysate sample to phosphorylate specific peptides imprinted on the PamChip® microarray (see Hilhorst R, Houkes L, van den Berg A, Ruijtenbeek R., “Peptide microarrays for detailed, high-throughput substrate identification, kinetic characterization, and inhibition studies on protein kinase A”, Anal. Biochem.

[0147] 2009;387:150-61). The PamChip® consists of four identical arrays, with each array containing 144 serine / threonine kinase (STK) or 196 protein tyrosine kinase (PTK) phosphosites. A detailed protocol is provided below:

[0148] Ear epidermal sections [60-pm combined from both ears per animal] were lysed in M-PER Mammalian Extraction Buffer (Thermo Fischer Scientific, # 78501) supplemented with Halt Phosphatase Inhibitor Cocktail (Thermo Fischer Scientific, # 78420) and Halt Protease Inhibitor Cocktail EDTA free (Thermo Fischer Scientific, # 87785). Following lysis, protein quantification was determined using Pierce Coomassie Plus (Bradford) Assay Kit (Thermo Fischer Scientific, #23236).

[0149] Measurements of kinome activity were performed on a PamStation-12 by PamGene (see Chirumamilla CS, Fazil M, Perez-Novo C, Rangarajan S, de Wijn R, Ramireddy P, et al., “Profiling activity of cellular kinases in migrating T-cells”, Methods Mol. Biol. 2019;1930:99-113). Briefly, the PamChip protein tyrosine kinase (PTK) array was processed in a single-step reaction in which five microgram of protein lysate was dispensed onto PTK array dissolved in protein kinase buffer and additives including 1% BSA, 10 mmol / L dithiothreitol, FITC conjugated pY20 antibody, and 400 mmol / L ATP. The STK array was processed in a two-step reaction in which 1 .5 microgram of protein lysate was used with protein kinase buffer supplied with 1% BSA, primary STK antibody mix, and 400 mmol / L ATP (sample mix). After an initial incubation of 110 minutes, the reaction mix was removed, and the secondary FITC-labeled antibody mix was added. In both arrays, software-based image analysis (BioNavigator software V.6.3 from PamGene) integrates the signals obtained within the time course of the incubation of the kinase lysate on the chip into one single value for each peptide for each sample (exposure time scaling). Peptide phosphorylation kinetics (for PTK) and its variations (for PTK / STK) were used to remove low signal peptides as quality control analysis (QC). Individual peptide phosphorylation intensities were transformed into Iog2. The peptides with significant differences in phosphorylation intensity (Treatment vs. control, using test, P < 0.05) are visualized as either volcano and bar plots or as heatmap using the R heatmap package.

[0150] Upstream kinase analysis (UKA) of PTK and STK data was done using default setting of the PamApp (PTK or STK UKA 2018 V.4.0) on BioNavigator Analysis software tool as described before (see Alack K, Weiss A, Kruger K, Horet M, Schermuly R, Freeh T, et al., “Profiling of human lymphocytes reveals a specific network of protein kinases modulated by endurance training status” Sci. Rep.

[0151] 2020; 10:888). The analysis is based on documented kinase-substrate relationships (from iviv database and literature-based protein modifications such as HPRD, PhosphoELM, PhosphositePLUS, Reactome, UNIPROT) complemented with “in silico” predictions that are retrieved from the phosphoNET database. The analysis generated three major parameters calculated by the PamApp: (i) Median kinase statistic (MKS), which depicts the overall change of the peptide set that represents a kinase. For example, a larger positive value indicates a larger activity in treated cells compared with controls, (ii) Mean significance score (MSiS), which indicates the significance of the change represented by the mean kinase statistic(s) between two groups (using 500 permutations across sample labels), (iii) Mean specificity score (MSpS), which indicates the specificity of the mean kinase statistics with respect to the number of peptides used for predicting the corresponding kinase (using 500 permutations across target peptides). The final ranking of the kinases was based on Median Final Score (MFS), which was calculated by addition of MSiS and MSpS. Top predicted kinases from significant (MFS>1.3) PTK and STK peptide sets are represented on phylogenetic tree of the human protein kinase family generated by Coral (see Metz KS, Deoudes EM, Berginski ME, Jimenez-Ruiz I, Aksoy BA, Hammerbacher J, Gomez SM, Phanstiel DH., “Coral: Clear and Customizable Visualization of Human Kinome Data”, Cell Syst. 2018;7(3):347-350).

[0152] Results

[0153] Changes in the median kinase score for various kinases after treatment with DD-L or control after day 1 can be seen in FIG 2. Kinases that showed a median kinase score of 1.3 or more 24 hours after treatment with DD-L were primarily tyrosine kinase (e.g., LYN, LCK, SRC, BLK, BMX, ERBB3, YES1, TEC, ABL1 and RYK). Serine / Threonine kinases were also observed to possess median kinase scores of 1.3 or more 24 hours after treatment with DD-L when compared to the kinase scores of the control (see FIG. 3). For example, AURKB, DAPK2, PRKCZ, PRKCB, PRKCD, PRKCQ, PRKCG, NPR2, CSNK1A, NEK8, and RPS6KA4 were observed to possess median kinase scores of 1.3 or more. After 48 hours of treatment with DD-L, serine / threonine kinases were still demonstrating median kinase scores of 1.3 or more (e.g., PRKAB1, MAP3K8, PRKAA1, ATR, STK38L, PRKY, CAMK2B, CSNK2A1, and PRKACG) (see FIG. 4). Serine / Threonine kinases were also observed to possess median kinase scores of 1.3 or more even after 72 hours of DD-L treatment (e.g., PRKG1 , PRKG2, PRKACA, CAMK2G, CAMK2D, RPS6KA5, DCLK2, CAMK2A, RPS6KB2, CSNK2A1, and CAMK4) (see FIG. 5). Serine / Threonine kinases were shown to possess median kinase scores of 1.3 or more after 48 hours of treatment with DD-I e.g., MAP3K8, ATR, CDK19, PRKACA, CAMK4, MAPKAPK2, and NEK8) (see FIG. 6). Serine / Threonine kinases also showed median kinase scores of 1.3 or more after 72 hours of treatment with DD-I (e.g., PRKCZ, MAP3K8, NUAK1, RPS6KA1, PRKCH, PRKCQ, DCLK2, PRKCI, AURKB and AURKA) (see FIG. 7).

[0154] Tyrosine kinases were shown to possess median kinase scores of 1.3 or more after 24 hours of treatment with DD-V when compared to the median kinase scores of the same tyrosine kinases after 24 hours of treatment with DMSO control (e.g., IGF1R, SRC, MST1R, LCK, INSR, FYN, ERBB4 and KDR) (see FIG. 8). Tyrosine kinases also demonstrated median kinase scores of 1.3 or more after 48 hours of treatment with DD-V when compared to those treated for 48 hours with DMSO control (e.g., FRK, LCK, SRMS, LYN, BLK, ZAP70, KDR, MST1R, FCGR3A, and CHEK1) (see FIG. 9). Tyrosine kinases were also observed to possess median kinase scores of 1.3 or more after 72 hours of treatment with DD-V when compared to those treated for 72 hours with DMSO control (e.g.,ERBB4, EPHA1, EGFR, RYK, FGR, NTRK2, NTRK3, NTRK1, INSRR, and HCK) (see FIG.

[0155] 10). Serine / Threonine kinases were observed to possess median kinase scores of 1.3 or more after 48 hours of treatment with DD-V when compared to those treated for 48 hours with DMSO control (e.g., PRKACA, CAMK4, STK38L, PIM1, MAP3K8, PRKACB, PRKG2, OAMK2B, OK17, and CSNK2A1) (see FIG. 11).

[0156] Serine / Threonine kinases also possessed median kinase scores of 1.3 or more after 72 hours of treatment with DD-V when compared to those treated for 72 hours with DMSO control (e.g., RPS6KB2, RPS6KA2, RPS6KA6, MYLK3, CHUK, DCLK2, DAPK3, MAPKAPK3, CHEK2, and PKN1) (see FIG. 12).

[0157] Example 2 Conclusion

[0158] The kinase activity profiling of the above exemplary PINK inhibitors demonstrates the ability of the inhibitors to inhibit that kinase activity of multiple tyrosine and serine / threonine kinases. Indeed, many kinases inhibited by DD-L, DD-I and DD-V have a pro-inflammatory function, thereby suggesting that each compound - DD-L, DD-I and DD-V have anti-inflammatory effect on psoriasis and other inflammatory skin diseases. This ability of the PINK inhibitors can play a critical role in the treatment of AD and psoriasis.

[0159] Example 2: Topical Treatment of Psoriasis and Atopic Dermatitis (AD) in Preclinical Models

[0160] Mouse Models

[0161] Eight- to 12-week-old female C57BL / 6J inbred mice used in this example were obtained from Jackson Laboratory (Bar Harbor, ME, USA). Mice were provided rodent chow and water ad libitum. The Penn State Institutional Animal Care and Use Committee (IACUC) approved all protocols involving mice (Protocol #: PRQT0202001655).

[0162] Treatment

[0163] Psoriasis was induced using 5% IMQ topically. The experimental procedure for inducing severe psoriasis in this Example is outlined in FIG. 13. Briefly, 5% IMQ was applied topically for 4 days, and treatment with DD-L, DD-I, or DD-V was started 2 days after IMQ treatment, when severe psoriasis developed, as evidenced by strongly increased skin thickness (see FIG. 14), and continued for 4 days, twice daily. The experimental procedure for inducing mild-to-moderate psoriasis is outlined in FIG. 15. Briefly, 5% IMQ was applied topically for 4 days, and treatment with DD-L, DD-I, or DD-V was started the same day after IMQ treatment, twice daily, starting 45 minutes post-IMQ treatment and continued for 4 days. Each group consisted of 5 mice. IMQ + / - treatment was applied to each ear (thus 10 controls and 10 treatments).

[0164] AD was induced using TPA (2 ug) once a day. Edema was induced on one or both ears of each C57BL / 6J mouse by the topical application of 12-0-Tetradecanoylphorbol-13-acetate (TPA, Sigma Chemical Co., U.S.A.) after dissolving it in DMSO at a concentration of 100 pg / ml. Each mouse was treated with 20pl (2 pg per ear) of TPA on the outer surface of an ear. The experimental procedure for inducing severe AD is outlined in FIG. 16. Briefly, 2% TPA was applied for 4 days, and treatment with DD-L, DD-I, or DD-V was started 2 days after TPA treatment, when severe eczema developed, as evidenced by strongly increased skin thickness (see FIG. 17), and continued for 4 days, twice daily. The experimental procedure for inducing mild-to-moderate eczema is outlined in FIG. 18. Briefly, 2% TPA was applied for 4 days, and treatment with DD-L, DD-I, or DD-V was started the same day after TPA treatment, twice daily, starting 45 minutes post-TPA treatment and continued for 4 days.

[0165] Affected skin was treated with: DMSO (control), 15% DD-L; 15% DD-I; or 15% DD-V. The therapeutic efficacy of DD-L, DD-I, and DD-V was measured by the changes in epidermal thickness assessed by micrometer (Mitutoyo Series IP65, Mitutoyo America, Aurora, IL) or by microscopy. Each group consisted of 5 mice. TPA + / - treatment was applied to each ear (thus 10 controls and 10 treatments).

[0166] Microscopy Based Measurement of Ear Thickness

[0167] Ear tissue (unfixed) was embedded in OCT and cryosectioned into 10 mm sections using a Leica CM 1850 cryostat. H&E-stained sections were imaged using a Zeiss AxioObserver 7.0 color camera. Average epidermal thickness was determined using Fiji software by drawing 10 lines perpendicular to the plane of the dorsal ear epidermis for each H&E image with lines spanning the innermost edge of the basal keratinocyte layer to the outermost edge of the granular keratinocyte layer. Images from at least 5 individual mice were quantified. Statistical analysis for ear thickness measurement by microscopy (see FIG. 19) or micrometer was done using nonparametric Student T test.

[0168] Results

[0169] DD-L, DD-I, and DD-V have Strong Therapeutic Effect in Preclinical Models of Severe Psoriasis and Severe Eczema

[0170] The therapeutic effect of PINK inhibitors was tested on preclinical models of severe psoriasis as described above. Results showed that treatment with DD-L, DD-I, and / or DD-V results in highly reduced skin thickness following induction of severe psoriasis (p<0.001) (see FIG. 20). This suggest that each of PINK inhibitors have strong therapeutic effect against severe psoriasis in preclinical model.

[0171] The therapeutic effect of PINK inhibitors was tested on preclinical models of severe AD as described above. Results showed that treatment with DD-L, DD-I, and / or DD-V results in reduced skin thickness following induction of severe eczema ( / .e., AD) (p<0.001) (see FIG. 21). This suggest that each of the PINK inhibitors have strong therapeutic effect against severe AD in preclinical model. DD-L showed the strongest therapeutic effect against severe AD among all three PINK inhibitors.

[0172] DD-L, DD-I, and DD-V have Strong Therapeutic Effect in Preclinical Models of Mild-to-Moderate Psoriasis and Mild-to-Moderate Eczema

[0173] The therapeutic effect of PINK inhibitors was tested on preclinical models of mild-to-moderate psoriasis and mild-to-moderate eczema as described above. Results showed that treatment with DD-L, DD-I, and / or DD-V results in highly reduced skin thickness following induction of mild-to-moderate psoriasis (p<0.001) (see FIG. 22). Results also showed that treatment with DD-L, DD-I, and / or DD-V results in reduced skin thickness following induction of mild-to-moderate eczema (p<0.001) (see FIG. 23). This suggests that each of PINK inhibitors have strong therapeutic effect against mild-to-moderate AD and mild-to-moderate psoriasis in preclinical model.

[0174] Novel PINK Inhibitors have Synergistic and / or Additive Therapeutic Effect with Hydrocortisone against Mild-to-Moderate Psoriasis and Mild-to-Moderate Atopic Dermatitis / Eczema

[0175] PINK inhibitors were evaluated for additive and / or synergistic therapeutic effects with hydrocortisone against mild-to-moderate psoriasis and mild-to-moderate eczema. Results showed that a topical treatment of mice with IMQ-induced mild-to-moderate psoriasis with hydrocortisone, as a single compound, (current standard treatment) has a therapeutic effect (see FIG. 24). However, treatment with a mixture of DD-L and 0.1% hydrocortisone or DD-V and 0.1% hydrocortisone had a significantly increased therapeutic effect on mild-to-moderate psoriasis, as compared to 0.1% hydrocortisone as a single drug (see FIG. 24). Similarly, treatment with mixture of DD-I and 0.25% hydrocortisone or DD-V and 0.25% hydrocortisone had a significantly increased therapeutic effect on mild-to-moderate psoriasis, as compared to 0.25% hydrocortisone as a single drug (see FIG. 25). These results demonstrate that the novel PINK inhibitors have an additive and / or synergistic therapeutic effect with hydrocortisone against mild-to moderate psoriasis and suggest that the dual treatment with these two drugs together is superior to the treatment with hydrocortisone alone. The PINK inhibitors were also tested for additive and / or synergistic therapeutic effects with hydrocortisone in a preclin ical model of mild-to-moderate atopic dermatitis / eczema. Treatment with 0.05%, 0.1% or 0.25% of hydrocortisone, showed a therapeutic effect against mild-to-moderate atopic dermatitis / eczema (see FIGs. 26-28). However, treatment with a mixture of DD-L, DD-I, or DD-V and hydrocortisone significantly increased the therapeutic effect on mild-to-moderate psoriasis (see FIGs. 26-28). This was observed when 0.05% hydrocortisone was used (see FIG. 26); 0.1% hydrocortisone was used (see FIG. 27), as well as when 0.25% hydrocortisone was used with DD-I or DD-V (see FIG. 28).

[0176] Additive or synergistic effects of PINK inhibitors with hydrocortisone were evidenced by a significant reduction of ear thickness following treatment with hydrocortisone + PINK inhibitors as compared to the treatment with Hydrocortisone alone. These results demonstrate that the PINK inhibitors have additive and / or synergistic therapeutic effects with hydrocortisone against mild-to moderate eczema and mild-to-moderate psoriasis and suggest that the dual treatment with these two drugs together is superior to the treatment with hydrocortisone alone.

[0177] It will be appreciated by those skilled in the art that the present disclosure can be embodied in other specific forms without departing from the spirit or essential characteristics thereof. For example, those skilled in the art will appreciate that multiple exemplary embodiments can be combined to produce embodiments falling within the scope of the disclosure that do not depart from the spirit of the disclosure, or the essential characteristics disclosed herein. The presently disclosed embodiments are therefore considered in all respects to be illustrative and not restricted. The scope of the disclosure is indicated by the appended claims rather than the foregoing description and all changes that come within the meaning and range and equivalence thereof are intended to be embraced therein.

Claims

CLAIMS:

1. A compound of Formula I:(Formula I)wherein:R1is selected fromR2is selected from -H, -CH3, -CH2CH3 and -CH(CH3)2;or a pharmaceutically acceptable salt or stereoisomer thereof.

2. The compound of claim 1, wherein R1is selected fromand3. The compound of claim 1 , wherein R1is selected from4. The compound of claim 3, wherein R2is selected from -H and -CH2CH3.

5. The compound of claim 3, wherein R2is CH2CH3.

6. A topical formulation comprising:a therapeutically effective amount of a compound of Formula I:(Formula I)wherein:R1is selected from; andR2is selected from -H, -CH3, -CFhCHsand -CH(CH3)2;or a pharmaceutically acceptable salt or stereoisomer thereof; and one or more cosmetic or pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic vehicles, water soluble vehicles, emulsifiers, nanoemulsifiers, buffers, humectants, moisturizers, solubilizers, preservatives, colorants, plasticizers, carriers, nanocarriers, dendrimers, liposomes, and / or excipients.

7. The topical formulation of claim 6, wherein the topical formulation is a lotion, solution, gel, an emulsion, microemulsion, a cream, an ointment, an aerosol, tapes, liposomal formulation or an ionic liquid formulation.

8. The topical formulation according to claim 6, comprising:one or more cosmetic peptides.

9. The topical formulation according to claim 6, comprising:one or more steroids.

10. The topical formulation according to claim 6, comprising:one or more natural anti-inflammatory agents.

11. The topical formulation according to claim 6, wherein the topical formulation contains from about 5 to about 30 wt% of the overall weight of the formulation of the compound of Formula I or a pharmaceutically acceptable salt or stereoisomer thereof.

12. A method for treating a skin condition in a subject in need thereof, comprising:administering to the subject in need of treatment for a skin condition a topical composition containing a therapeutically effective amount of a compound of Formula I:(Formula I) wherein:R1is selected fromR2is selected from -H, -CH3, -CH2CH3and -CH(CH3)2;or a pharmaceutically acceptable salt or stereoisomer thereof; and one or more cosmetic or pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic vehicles, water soluble vehicles, emulsifiers, nano-emulsifiers, nano-suspension, buffers, humectants, moisturizers, solubilizers, preservatives, colorants, plasticizers, carriers, nano-carriers, dendrimers, liposomes, and / or excipients.

13. The method of claim 12, wherein the skin condition is characterized by one or more of inflammation, itching, redness, rapid skin growth, thickening of skin or blistering of the skin.

14. The method of claim 12, wherein R1is selected from15. The method of claim 12, wherein R1is selected fromsr<A.Art / vw'and16. The method of claim 15, wherein R2is selected from -H and -CH2CH3.

17. The method of claim 16, wherein R2is CH2CH3.

18. A method for treating a malignant disease or autoimmune disorder in a subject in need thereof, comprising:administering to the subject in need of treatment for a malignant disease or autoimmune disorder a composition containing a therapeutically effective amount of a compound of Formula I:(Formula I)wherein:R1is selected from5; andR2is selected from -H, -CH3, -CFLCHsand -CH(CHs)2;or a pharmaceutically acceptable salt or stereoisomer thereof.

19. The method of claim 18, wherein the composition is administered systemically or topically to the subject in need thereof.

20. The method of claim 18, wherein the malignant disease is a cancer.

21. The method of claim 18, wherein the autoimmune disorder is selected from mild-to-server atopic dermatitis (eczema), mild-to-severe psoriasis, Lupus, dermatomyositis, Lichen Planus, Pemphigus, hidradenitis suppurativa, Vitiligo, Scleroderma, and Pemphigoid.

22. The method of claim 18, comprising:administering a steroid with or after administration of the composition.

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